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Distant organ crosstalk during AKI, first studied mechanistically in the lung (1), is now known to play an important role in disease outcomes. The relationship between intestinal microbiota and AKI has also been explored, resulting in exciting findings that are promising for future human therapeutics. In search of ways to reduce kidney T cells by using “cleaner,” germfree (GF) mice, it was unexpectedly found that GF mice not only retained their kidney immune cells, but also had a higher number of natural killer T (NKT) cells and reduced IL-4 levels compared with normal wild-type (WT) mice. Moreover, GF mice subjected to kidney ischemia-reperfusion (IR) injury had increased CD8 T-cell trafficking and inflammatory cytokine mediators compared with normal WT mice (2). Induction of ischemic AKI in GF mice led to a worse course of AKI, in terms of both structure and function. Conventionalizing GF mice with normal mouse stool led to normalizing T-cell and NKT populations, and relative protection from AKI, compared with GF mice (2). It was subsequently observed that early exposure to gut microbiota during development was associated with long-term changes in NKT-cell function in nonrenal tissues, and NKT cells in GF mice had a less mature phenotype with diminished activation capacity upon antigen encounter (3,4). To elucidate whether the effects were bidirectional and AKI led to changes in the gut microbiome, gut microbiota was evaluated in C57BL/6 WT mice after ischemic or nephrotoxic (cisplatin; CP) injury. DNA was isolated from the fecal pellets ( n =4–5 per group) at baseline (D0) and at 3 days (D3) post-IR. The V3–V5 region of the 16S ribosomal RNA gene was amplified using the 357F/926R primer set and sequenced using the Roche/454 platform. IR and CP modified the relative abundance of specific bacterial species belonging to the phyla Actinobacteria, Bacteroidetes, Firmicutes, …

We investigated the clinical relevance of urinary cytokines/chemokines reflecting intrarenal immunologic micromilieu as prognostic markers and the optimal measurement timing after living donor kidney transplantation (LDKT). This prospective cohort study included 77 LDKT patients who were followed for ≥ 5 years. Patients were divided into control (n = 42) or acute rejection (AR, n = 35) group. Early AR was defined as AR occurring within 3 months. Serum and urine cytokines/chemokines were measured serially as follows: intraoperative, 8/24/72 h, 1 week, 3 months, and 1 year after LDKT. Intrarenal total leukocytes, T cells, and B cells were analyzed with immunohistochemistry followed by tissueFAXS. Urinary MCP-1 and fractalkine were also analyzed in a validation cohort. Urinary MCP-1 after one week was higher in the AR group. Urinary MCP-1, fractalkine, TNF-α, RANTES, and IL-6 after one week were significantly higher in the early AR group. Intrarenal total leukocytes and T cells were elevated in the AR group compared with the control group. Urinary fractalkine, MCP-1, and IL-10 showed positive correlation with intrarenal leukocyte infiltration. Post-KT 1 week urinary MCP-1 showed predictive value in the validation cohort. One-week post-KT urinary MCP-1 may be used as a noninvasive diagnostic marker for predicting AR after LDKT.

The versatility of the intrarenal immunologic micromilieu through dietary modification and the subsequent effects on susceptibility to ischemic acute kidney injury (AKI) are unclear. We investigated the effects of high-salt (HS) or high-fat (HF) diet on intrarenal immunologic micromilieu and development of ischemic AKI using murine ischemic AKI and human kidney-2 (HK-2) cell hypoxia models. Four different diet regimens [control, HF, HS, and high-fat diet with high-salt (HF+HS)] were provided individually to groups of 9-week-old male C57BL/6 mice for 1 or 6 weeks. After a bilateral ischemia-reperfusion injury (BIRI) operation, mice were sacrificed on day 2 and renal injury was assessed with intrarenal leukocyte infiltration. Human kidney-2 cells were treated with NaCl or lipids. The HF diet increased body weight and total cholesterol, whereas the HF+HS did not. Although the HF or HS diet did not change total leukocyte infiltration at 6 weeks, the HF diet and HF+HS diet increased intrarenal CD8 T cells. Plasma cells increased in the HF and HS diet groups. The expression of proinflammatory cytokines including TNF-α, IFN-γ, MCP-1, and RANTES was increased by the HF or HS diet, and intrarenal VEGF decreased in the HS and HF+HS diet groups at 6 weeks. Deterioration of renal function following BIRI tended to be aggravated by the HF or HS diet. High NaCl concentration suppressed proliferation and enhanced expression of TLR-2 in hypoxic HK-2 cells. The HF or HS diet can enhance susceptibility to ischemic AKI by inducing proinflammatory changes to the intrarenal immunologic micromilieu.

Intrarenal robust inflammatory response following ischemia-reperfusion injury (IRI) is a major factor in the pathogenesis of renal injury in ischemic acute kidney injury (AKI). Although numerous studies have investigated various agents of immune modulation or suppression for ischemic AKI, few showed reproducible effects. We hypothesized that poly (ADP-ribose) polymerase (PARP) inhibitor may favorably change post-ischemic intrarenal immunologic micromilieu by reducing damage-associated molecular pattern (DAMP) signals and improve renal outcome in ischemic AKI. The effects of JPI-289 (a PARP inhibitor) on early renal injury in a murine IRI model and hypoxic HK-2 cell model were investigated. Bilateral IRI surgery was performed in three groups of 9-week-old male C57BL/6 mice (control, JPI-289 50 mg/kg, and JPI-289 100 mg/kg; n = 9-10 in each group). Saline or JPI-289 was intraperitoneally injected. Renal function deterioration was significantly attenuated in the JPI-289 treatment groups in a dose-dependent manner. Inflammatory cell infiltration and proinflammatory cytokine/chemokine expressions in the post-ischemic kidneys were also attenuated by JPI-289 treatment. JPI-289 treatment at 0.5 and 0.75 μg/ml facilitated the proliferation of hypoxic HK-2 cells. PARP inhibition with JPI-289 treatment showed favorable effects in ischemic AKI by attenuating intrarenal inflammatory cascade in a murine model and facilitating proliferation of hypoxic HK-2 cells.

Background: Patients undergoing kidney transplantation (KT) often receive red blood cell (RBC) transfusion during admission for KT which may increase the risk of allosensitization. The association between peri-transplant RBC transfusion and graft survival was evaluated using a nationwide cohort. Methods: This retrospective study analyzed 13,871 patients who underwent KT in Korea between 2007 and 2015. The outcomes were graft failure rate and overall patient survival depending on the amount of RBC transfusion. Results: The overall graft failure rate was 15.5%. Compared to the graft failure rate of 13.5% in the no transfusion group, the graft failure rate was 15.4% in the 1–2 units group (sHR 1.06 (95% CI 0.97–1.17), p = 0.216), 21.4% in the 3–5 units group (sHR 1.39 (1.21–1.61), p &lt, 0.001), and 35.3% in the 6 or more units group (sHR 2.20 (1.70–2.85), p &lt, 0.001). The overall survival rate was 97.5% in the no transfusion group, compared to 95.9% in the 1–2 units group (HR 1.50 (1.22–1.83), p &lt, 0.001), 92.0% in the 3–5 units group (HR 2.43 (1.87–3.15), p &lt, 0.001), and 67.5% in the 6 or more units group (HR 6.81 (5.03–9.22), p &lt, 0.001). Conclusions: Peri-transplant RBC transfusion was independently associated with the increased risk of renal allograft failure and death in KT patients.

Hydroxypropyl cyclosophoraose-pullulan (HPCys-Pul) microspheres were designed as a novel hybrid system of biocompatible pullulan matrice and pendant hydroxypropyl cyclosophoraoses with drug complexing ability. The HPCys-Pul microspheres were prepared by emulsion crosslinking method, and characterized using thermogravimetric analysis, Fourier transform infrared spectroscopy, scanning electron microscopy, and solid state NMR spectroscopy. By virtue of hydroxypropyl cyclosophoraoses, the developed microspheres can encapsulate 4.2-fold more naproxen than pullulan microspheres. Korsmeyer–Peppas model was introduced for describing release kinetics. In vivo naproxen release analysis was carried out in Sprague-Dawley (SD) rats. From those results HPCys-Pul microsphere will be a promising platform for controlled drug delivery.

Ultraviolet radiation commonly causes serious skin diseases, and skin cell death. The UVB-blocking effect of hemp fabric which known to be a powerful agent against UVB was evaluated using mouse auricle skin. Based on UVB irradiation and the use of different fabrics, four mouse groups were evaluated in this study: two experimental groups, Group 1 (UVB exposed hemp fabric-shield site, EHFS), Group 2 (UVB exposed polyester fabric-shield site, EPFS), and two control groups, Group 3 (UVB exposed non-fabric-shield site, ENFS), and Group 4 (UVB unexposed non-fabric-shield site, UNFS). Except for UNFS all samples were exposed to UVB for 28 h and showed clear histologic changes in epidermis and dermis. After 45 h chronic irradiation, epidermal thickness was doubled in EHFS, roughly tripled in EPFS, and more than quadrupled in ENFS over that of UNFS. Based on the thickness of the altered epidermis, the blocking effect of hemp fabric was 50% higher than that of polyester fabric. Additionally, immunohistochemical analysis revealed expression of proliferating cell nuclear antigen (PCNA) in ENFS and EPFS throughout the hyperplasia of keratinocytes and sebocytes. After 45 h irradiation, the sebocytes of sebaceous glands, observed in sectioned images, increased on average to 14 cells in ENFS, 9 cells in EPFS, and 7 cells in EHFS, as compared to 8 cells in UNFS. In contrast, the cell area of adipose tissue was decreased by half in EHFS, one-fourth in EPFS, and one-tenth in ENFS, and mostly replaced with fibroblasts and other supporting cells. These results suggest that UVB irradiation directly affects epidermal and dermal tissues, and induces abnormal proliferation of keratinocytes and hyperplasia of sebocytes consuming fats in adipose tissue. For skin health, hemp fabric is a better material for protecting the skin against UVB than polyester fabric.

Luteolin [2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4-chromenone] is an active flavonoid compound from Lonicera japonica (Caprifoliaceae). Luteolin inhibits tumor cell proliferation, inflammatory and oxidative stress better, when compared with other flavonoids. In the present study, it was demonstrated that luteolin induces typical apoptosis in PC12 cells (derived from a pheochromocytoma of the rat adrenal medulla) accompanied by DNA fragmentation and formation of apoptotic bodies. In addition, luteolin regulates expression of the endoplasmic reticulum (ER) chaperone binding immunoglobulin protein, activating ER stress sensors (eukaryotic initiation factor 2α phosphorylation and X‑box binding protein 1 mRNA splicing) and induced autophagy. The results indicated that luteolin induces the upregulation of the unfolded protein response pathway through the ER stress sensors, which helps as an influential regulator for the apoptosis pathway in PC12 cells. The results suggested that the understanding of the molecular mechanisms underlying luteolin‑induced apoptosis may be useful in cancer therapeutics, chemoprevention and neurodegenerative diseases, such as Parkinson's disease and Alzheimer's disease.

The Down syndrome critical region 1 (DSCR1), a Down syndrome-associated protein, is an endogenous inhibitor of the Ca2+-dependent phosphatase calcineurin. It has been also suggested to be associated with Alzheimer’s disease (AD) but the role of DSCR1 in the pathogenesis of AD still remains controversial. In this paper, we investigated the effects of knockdown of sarah (sra), a Drosophila DSCR1 ortholog, on the Aβ42-induced developmental phenotypes of Drosophila. Knockdown of sra showed detrimental effects on the rough eye phenotype and survival of Aβ42-expressing flies without altering the Aβ42 accumulation. Furthermore, the knockdown of sra increased glial cell numbers in the larval brains and its susceptibility to oxidative stress. Overexpression of an active form of calcineurin produced similar results to sra knockdown as they both exacerbated the Aβ42-induced rough eye phenotype. However, sra knockdown did not alter apoptosis or c-Jun N-terminal kinase activation in Aβ42-expressing flies. In conclusion, our results suggest that sra does play an important role in Aβ42-induced developmental defects in Drosophila without affecting its stress responses.

A methoxylated flavanone derivative, 2′,3′,4′-trimethoxy-5,6-naphthoflavanone (TMNF), was synthesized, and its anti-proliferative activity was evaluated. In HCT116 colon cancer cells, TMNF inhibited cellular proliferation and triggered cytotoxicity. A flow cytometry assay showed that TMNF caused cell cycle arrest at the G2/M phase with a concomitant decrease in the population of G1 phase cells. Furthermore, TMNF caused accumulation of p53 and its downstream target, Bax, and stimulated cleavage of caspase-7 and poly(ADP-ribose) polymerase. These data suggest that methoxylated naphthoflavanone can be an attractive pharmacophore for further development of therapeutic adjuvants.

Eukaryotic translation initiation factor 2 alpha (eIF2α), which is a component of the eukaryotic translation initiation complex, functions in cell death and survival under various stress conditions. In this study, we investigated the roles of eIF2α phosphorylation in cell death using the breast cancer cell lines MCF-7 and MCF-7/ADR. MCF-7/ADR cells are MCF-7-driven cells that have acquired resistance to doxorubicin (ADR). Treatment of doxorubicin reduced the viability and induced apoptosis in both cell lines, although susceptibility to the drug was very different. Treatment with doxorubicin induced phosphorylation of eIF2α in MCF-7 cells but not in MCF-7/ADR cells. Basal expression levels of Growth Arrest and DNA Damage 34 (GADD34), a regulator of eIF2α, were higher in MCF-7/ADR cells compared to MCF-7 cells. Indeed, treatment with salubrinal, an inhibitor of GADD34, resulted in the upregulation of eIF2α phosphorylation and enhanced doxorubicin-mediated apoptosis in MCF-7/ADR cells. However, MCF-7 cells did not show such synergic effects. These results suggest that dephosphorylation of eIF2α by GADD34 plays an important role in doxorubicin resistance in MCF-7/ADR cells.

Purpose The incidence of chronic kidney disease (CKD) has been increasing due to improved survival after liver transplantation (LT). Risk factors of kidney injury after LT, especially perioperative management factors, are potentially modifiable. We investigated the risk factors associated with progressive CKD for 10 years after LT. Methods This retrospective cohort study included 292 adult patients who underwent LT at a tertiary referral hospital between 2000 and 2008. Renal function was assessed by the e stimated glomerular filtration rate (eGFR) using the Modification of Diet in Renal Disease formula. The area under the curve of serial eGFR (AUCeGFR) was calculated for each patient to assess the trajectory of eGFR over the 10 years. Low AUCeGFR was considered progressive CKD. Linear regression analyses were performed to examine the associations between the variables and AUCeGFR. Results Multivariable analysis showed that older age (regression coefficient = −0.53, P < 0.001), diabetes mellitus (DM) (regression coefficient = −6.93, P = 0.007), preoperative proteinuria (regression coefficient = −16.11, P < 0.001), preoperative acute kidney injury (AKI) (regression coefficient = −14.35, P < 0.001), postoperative AKI (regression coefficient = −3.86, P = 0.007), and postoperative mean vasopressor score (regression coefficient = −0.45, P = 0.034) were independently associated with progressive CKD. Conclusion More careful renoprotective management is required in elderly LT patients with DM or preexisting proteinuria. Postoperative AKI and vasopressor dose may be potentially modifiable risk factors for progressive CKD.

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease characterized by synovitis, hyperplasia, and the destruction of bone and cartilage. A variety of immunosuppressive biological agents have been developed because the pathogenesis of RA is related predominantly to the inflammatory response. However, rheumatoid arthritis fibroblast-like synovial cells (RAFLS), which are known to play an important role in RA progression, exhibit resistance to immunosuppressants through cancer-like properties. In this study, we identified a novel therapeutic compound for RA, which reduced inflammation and the abnormal proliferation of RAFLS in natural product library made from Korean native plants. Eupatorium japonicum Thunb. (EJT) extract, a component of the natural product library, most effectively reduced viability through the induction of ROS-mediated apoptosis in a dose-dependent manner. In addition, the increased ROS induced the expression of ATF4 and CHOP, key players in ER stress-mediated apoptosis. Interestingly, EJT extract treatment dose-dependently reduced the expression of IL-1β and the transcription of MMP-9, which were induced by TNF-α treatment, through the inhibition of NF-κB and p38 activation. Collectively, we found that EJT extract exerted apoptotic effects through increases in ROS production and CHOP expression and exerted anti-inflammatory effects through the suppression of NF-κB activation, IL-1β expression, and MMP-9 transcription.

Alzheimer's disease (AD) is the most common neurodegenerative disorder, characterized by progressive neuronal loss with amyloid β-peptide (Aβ) plaques. Despite several drugs currently used to treat AD, their beneficial effects on AD progress remains under debate. Here, we established a rapid in vivo screening system using Drosophila AD models to assess the neuroprotective activities of medicinal plants that have been used in traditional Chinese medicine. Among 23 medicinal plants tested, the extracts from five plants, Coriandrum sativum, Nardostachys jatamansi, Polygonum multiflorum (P. multiflorum), Rehmannia glutinosa, and Sorbus commixta (S. commixta), showed protective effects against the Aβ42 neurotoxicity. We further characterized the neuroprotective activity of ethanol extracts from P. multiflorum and S. commixta. Aβ42-expressing flies that we used showed AD neurological phenotypes, such as decreased survival and motility and increased cell death and reactive oxygen species level. However, feeding these flies extracts from P. multiflorum or S. commixta showed strong suppression of such phenotypes. Similar results were observed in human cells, so that the treatment of P. multiflorum and S. commixta extracts increased the viability of Aβ-treated SH-SY5Y cells. Moreover, 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside, one of the main constituents of P. multiflorum, also showed similar protective activity against Aβ42 cytotoxicity in both Drosophila and human cells. Taken together, our results suggest that both P. multiflorum and S. commixta have therapeutic potential for the treatment of neurodegenerative diseases, such as AD.

Letter to the Editor

BACKGROUND IgA nephropathy (IgAN) is the most frequent primary glomerular disease and the leading cause of end-stage renal disease. We investigated clinicopathologic predictors of renal survival in patients with IgAN with a focus on glomerular filtration rate (GFR) decline slope. MATERIALS AND METHODS We screened all patients with primary IgAN between 1995 and 2012. Renal progression was defined as doubling of serum creatinine. Using serial serum creatinine levels during the first-year, we calculated the GFR decline slopes. Further, we defined patients in the steepest GFR slope quartile as rapid decliners and those in the second steepest GFR slope quartile as slow decliners. Others were defined as nondecliners. RESULTS Of 214 participants, baseline GFR was 81 (62, 100) mL/min/1.73 m2 , which was not different among the 3 groups. Rapid decliners and slow decliners had higher levels of urinary protein/creatinine ratio (0.88, 0.89 and 0.58 g/gCr, respectively, P

The importance of forests continues to increase throughout the world, and one of the reasons is that a forest is a major place to emit terpenes, which have been reported to be beneficial to human health. In South Korea, forests occupy about 64% of the total land area and consist mainly of pine and oak trees. Since only a limited number of forests have been analyzed to date, a comprehensive understanding of terpenes emitted from regional forests remains in its infancy in Korea. Here, to gain insights into terpenes from regional forests located in South Korea, we review the characteristics of Korean forests and recent studies on major terpenes emitted from regional forests as well as from native trees dominant in South Korea. We also discuss meteorological factors that affect the terpene emissions in Korean forests. In conclusion, 18 types of terpenes were detected in Korean forests and their compositions in different forests are largely dependent on the dominant plant species in the forest. Moreover, terpene emissions in Korean forests are affected by various environmental factors, including temperature, amount and duration of daylight, season, and age of trees. To improve the understanding of the characteristics of terpene distribution, more studies are required on the terpene production of Korean forests in various regions.

Ethnopharmacological relevance The medicinal efficacy of hempseed (Cannabis sativa L.), which is rich in polyunsaturated fatty acids, in atopic dermatitis, inflammation, and rheumatoid arthritis (RA) has been suggested for centuries. Hempseed has been used as a treatment for these diseases in Korean and Chinese folk medicine. The aim of the study is to investigate the effects of hempseed oil (HO) on MH7A human RA fibroblast-like synovial cells. Materials and methods MH7A cells were used to study the anti-rheumatoid effects of hempseed (Cannabis sativa L., cv. Cheungsam/Cannabaceae) oil by investigating cell viability, apoptosis, lipid accumulation, oxidative stress, and endoplasmic reticulum (ER) stress-induced apoptosis. Results HO treatment reduced the survival rate of MH7A cells and promoted apoptotic cell death in a time- and dose-dependent manner. Both lipid accumulation and the level of intracellular reactive oxygen species (ROS) increased in HO-treated MH7A cells. Co-treatment with the antioxidant Tiron effectively abrogated the cytotoxic effects of HO; the ROS level was reduced, cell viability was recovered, and apoptotic cell death was significantly diminished. Moreover, HO-treated cells exhibited increased expression of the major ER stress markers, glucose-regulated protein 78 and C/EBP homologous protein (CHOP). The siRNA-mediated knockdown of CHOP prevented HO-induced apoptosis. Conclusions Our results suggest that HO treatment induced lipid accumulation, ROS production, CHOP expression, and apoptosis in MH7A cells, and that CHOP functions as an anti-rheumatoid factor downstream of HO in MH7A cells.

Plasma cells are exposed to intense endoplasmic reticulum (ER) stress imposed by enormous rates of immunoglobulin (Ig) synthesis and secretion. Therefore, protein homeostasis is crucial for the survival of plasma cells, but its molecular mechanism remains largely unknown. Here we found marked overexpression of FK506-binding protein 13 (FKBP13) in long-lived plasma cells from autoimmune mice, and investigated its function using a plasmacytoma cell line secreting IgA. FKBP13 expression was induced largely in the lumen of ER in response to treatment with an ER stressor tunicamycin or overexpression of an adaptive unfolded protein response protein X-box binding protein 1 (XBP1). Silencing of FKBP13 expression led to induction of molecules involved in the terminal unfolded protein response and ER stress-associated apoptosis. FKBP13 interacted with Ig, facilitated its ubiquitination, and lowered the extent of ER stress. FKBP13 overexpression caused a significant reduction in secreted IgA in plasmacytoma cells, and FKBP13 knockdown exerted an opposite effect. Rapamycin interfered with the interaction between FKBP13 and IgA and enhanced the amount of secreted IgA. Importantly, the level of FKBP13 was inversely correlated with the amount of secreted antibody in long-lived plasma cells from autoimmune mice. These results suggest that FKBP13 is a marker of long-lived plasma cells and a component of XBP1-dependent ER protein homeostasis. FKBP13 is likely to act as a molecular chaperone that delivers misfolded ER clients, including Ig, to ER-associated degradation, so reducing proteotoxic stress on the plasma cell. Our data reveal a novel cytoprotective role for FKBP13 in long-lived plasma cells occurring at the expense of antibody production.

The purpose of this study was to determine the late decline in viability of fat cells over time for fat tissue stored at -15 degrees C and -70 degrees C after harvest from abdominal liposuction. A total of 16 females were recruited for this study. The viability of fat cell specimens was measured after freezing for 1, 3, 7, 14, 28, and 56 days. A number of viable mature adipocytes were evaluated by fluorescence microscopy after staining with fluorescein diacetate and propidium iodide. The glycerol-3-phosphate dehydrogenase activity was measured in lipoaspirates before digestion and the XTT reduction assay was performed. In addition, the XTT reduction assay was also performed on isolated lipocytes and preadipocytes.The viability of mature adipocytes was very low for both the -15 degrees C and -70 degrees C samples after 1 day of freezing (13.3% +/- 7.4% and 12.6% +/- 6.3%, respectively). There was no statistically significant difference between the samples stored at the 2 temperatures. The GPDH activity of the lipoaspirates frozen, for 1 day, at -15 degrees C and -70 degrees C was 25.1% +/- 10% and 28.7% +/- 11%, respectively. For the XTT test, the fractional enzyme activity of the lipoaspirates frozen, for 1 day, at -15 degrees C and -70 degrees C was 30.0% +/- 10.9% and 36.1% +/- 12.3%, respectively. In addition, the adipocytes had low activity from day one: 15.4% +/- 7.2% at -15 degrees C and 11.5% +/- 5.6% at -70 degrees C. Furthermore, the preadipocytes had a low activity of 8.0% +/- 6.0% at -15 degrees C and 8.6% +/- 3.8% at -70 degrees C. At 8 weeks, there were few viable mature adipocytes and the activity of the cells was very low by XTT and GPDH testing.The results of this study showed that the viability of adipocytes declined rapidly after frozen storage for 1 day at both -15 degrees C and -70 degrees C, and decreased gradually in storage after 8 weeks; at which time only approximately 5% of the fat cells were alive. These findings suggest that the present fat preservation storage techniques using a -15 degrees C freezer or a -70 degrees C deep freezer are both inadequate to maintain the viability of fat cells.

Fibroblast-like synovial cells play a crucial role in the pathophysiology of rheumatoid arthritis (RA), as these cells are involved in inflammation and joint destruction. Apigenin, a dietary plant-flavonoid, is known to have many functions in animal cells including anti-proliferative and anticancer activities, but its role in human rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) has not been reported. In this study, we investigated the roles of apigenin in RA-FLSs. The survival rate decreased, and apoptotic cell death was induced by apigenin treatment in RA-FLSs. Apigenin treatment resulted in activation of the mitogen-activated protein kinase (MAPK) ERK1/2, and pretreatment with an ERK inhibitor PD98059 dramatically reduced apigenin-induced apoptosis. We found that apigenin-mediated production of a large amount of intracellular reactive oxygen species (ROS) caused activation of ERK1/2 and apoptosis; treatment with the antioxidant Tiron strongly inhibited the apigenin-induced generation of ROS, phosphorylation of ERK1/2, and apoptotic cell death. Apigenin-induced apoptotic cell death was mediated through activation of the effectors caspase-3 and caspase-7, and was blocked by pretreatment with Z-VAD-FMK (a pan-caspase inhibitor). These results showed that apigenin-induced ROS and oxidative stress-activated ERK1/2 caused apoptotic cell death in apigenin-treated RA-FLSs.

Macrophages secrete inflammatory cytokines and mono-nitrogen oxide (NO), and play crucial roles in inflammation in early-stage rheumatoid arthritis (RA). This study investigated whether glucosamine hydrochloride (GlcN), a nonsteroidal anti-inflammatory agent (NSAID) widely used to treat arthritis, affects the expression of inflammatory cytokines via the unfolded protein response (UPR) in lipopolysaccharide (LPS)-stimulated mouse macrophages (RAW264.7 cells). Pretreatment with GlcN reduced the expression of inflammatory cytokines and inhibited cell differentiation. Moreover, GlcN treatment increased the expression of CHOP and BiP/Grp78, the UPR target genes, in the presence or absence of LPS. Indeed, knockdown of CHOP using siRNAs prevented the GlcN-mediated reduction of inflammatory cytokines in LPS-stimulated RAW264.7 cells. Finally, we found that GlcN-mediated induction of CHOP reduced the phosphorylation of JNK and NF-?B in LPS-stimulated RAW264.7 cells. Combined, these results suggest that the GlcN-mediated induction of CHOP negatively regulates the inflammatory response by modulating JNK and NF-?B in LPS-stimulated RAW264.7 cells.

Micro-deletions at specific loci of the Y chromosome have been observed frequently in male infertility patients, suggesting that genes in these regions are involved in male germ cell development. DAZ is a representative male infertility gene at the AZFc locus of the Y chromosome. Since DAZ contains an RNA binding motif along with so-called a DAZ domain, it was proposed to participate in RNA metabolism during spermatogenesis. A mouse gene homologous to the human DAZ gene has been cloned and named Dazl (DAZlike). Dazl is autosomal and expressed in the testis and also at a low level in the ovary. Male mice homozygous for the Dazl null allele have small testes with a few spermatogonia and almost complete absence of germ cells beyond the spermatogonial stage, suggesting the requirement of Dazl for entry or progression through meiosis. However, its exact cellular functions have not been understood yet. In order to investigate cellular functions of Dazl, we decided to isolate candidate interacting protein genes of the mouse Dazl, using yeast two-hybrid screening. A number of candidate Dazlinteracting proteins have been isolated, such as Bprp, Acf, Hgs, Murr1, Nbak3 and Ranbp9, but dynein light chain 1 (Dlc1) was most predominant. A strong interaction of Dazl with Dlc1 suggests that Dazl might function as an mRNA adaptor to the dynein motor complex.

Background: Codman's paradox reveals a misunderstanding of geometry in orthopedic practice. Physicians often encounter situations that cannot be understood intuitively during orthopedic interventions such as corrective osteotomy. Occasionally, unexpected angular or rotational deformity occurs during surgery. This study aimed to draw the attention of orthopedic surgeons toward the concepts of orientation and rotation and demonstrate the potential for unexpected deformity after orthopedic interventions. This study focused on three situations: shoulder arthrodesis, femoral varization derotational osteotomy, and femoral derotation osteotomy. Methods: First, a shoulder model was generated to calculate unexpected rotational deformity to demonstrate Codman's paradox. Second, femoral varization derotational osteotomy was simulated using a cylinder model. Third, a reconstructed femoral model was used to calculate unexpected angular or rotational deformity during femoral derotation osteotomy. Results: Unexpected external rotation was found after forward elevation and abduction of the shoulder joint. In the varization and derotation model, closed-wedge osteotomy and additional derotation resulted in an unexpected extension and valgus deformity, namely, under-correction of coxa valga. After femoral derotational osteotomy, varization and extension of the distal fragment occurred, although the extension was negligible. Conclusions: Surgeons should be aware of unexpected angular deformity after surgical procedure involving bony areas. The degree of deformity differs depending on the context of the surgical procedure. However, this study reveals that notable deformities can be expected during orthopedic procedures such as femoral varization derotational osteotomy.

Essential hypertension is a multifactorial disease, and has been shown to be associated with insulin resistance. The relationship between the genetic variation of the insulin receptor (INSR) gene and essential hypertension in Korean population was investigated by the Nsi I restriction fragment length polymorphism (RFLP) pattern of this gene. The observed genotype frequencies of INSR gene were not deviated from those expected for the Hardy‐Weinberg equilibrium (HWE), but a significant association was observed between essential hypertension and N1 allele of Nsi I RFLP at the INSR gene (χ2‐test; P

Enteropathy-associated T-cell lymphoma (EATL) is a rare extranodal T-cell lymphoma arising from the intestine. Two types of EATL have been reported. In contrast to the classic EATL type I, EATL type II occurs sporadically, is unrelated to celiac disease, and comprises 10% to 20 % of all EATL cases. A total of five cases of EATL type II were diagnosed at our clinic from January 2009 to September 2012. Four of the five patients were diagnosed with the help of endoscopy. Among the four patients, two of the cases involved both the small and large intestines, whereas in the other two patients, EATL was limited to the small intestine. Common endoscopic findings included innumerable fine granularities (also called mosaic mucosal patterns) and diffuse thickening of the mucosa with a semicircular shallow ulceration in the lesions of the small bowel. In contrast, the endoscopic findings of the colon were nonspecific and could not distinguish EATL type II from other diseases. There are only few published reports regarding the representative endoscopic findings of EATL. Here, we present the clinical and endoscopic findings of four cases of EATL type II diagnosed by endoscopy.

본 증례에서 저자들은 단백뇨와 신기능 저하를 보이는 당뇨 환자에서 당뇨 망막 병증이 동반되어 있고 수 년간 혈당조절 없이 지냈던 점을 감안하여 당뇨 신 질환을 의심했다. 그러나 단백뇨의 주요 성분이 알부민이 아니고 혈청 면역글로불린 농도의 저하가 동반되어 있어 혈청, 요 면역 전기영동 검사와 신생검을 시행한 결과 경쇄 침착 질환을 진단하고 화학 요법과 자가 조혈모세포 이식을 통해 효과적으로 치료한 증례를 경험하였다. 당뇨가 있으면서 단백뇨와 신부전을 보이는 환자에서 비전형적인 임상양상을 보이는 경우에는 적극적으로 신조직 검사를 고려해야 하며, 당뇨 신 질환과 유사한 광학현미경 소견을 보이는 경쇄 침착 질환의 감별을 위해서는 경쇄에 대한 면역형광 염색과 전자현미경소견에 대한 주의 깊은 관찰이 필요하겠다.

Delayed perforation is a very rare complication of endoscopic submucosal dissection (ESD), with a reported incidence of 0.1% to 0.45%. Few reports exist on the clinical features and outcomes of delayed perforation after ESD, and it is unclear whether the optimal management strategy is emergency surgery or endoscopic closure with conservative treatment. Here, we report two cases of delayed perforation occurring after ESD for early gastric cancer. In both cases, lesions were located in the antrum, and tumor depths were confined to the mucosal layer. Total procedure times for ESD were 25 and 45 minutes, respectively. Because delayed perforation may be associated with excessive thermal damage and necrosis of the muscle layer, treatment with emergency surgery should be used instead of conservative management in cases of delayed perforation after ESD.

To the Editor, An adrenal "incidentaloma" is an adrenal mass, ≥ 1 cm diameter, that is discovered by chance during a radiological examination performed for medical conditions other than adrenal disease [1]. Clinically, about 80% of adrenal incidentalomas are nonfunctional, benign adrenocortical adenomas, 9% cause subclinical Cushing's syndrome, 4% are pheochromocytomas, and only 1% are aldosteronomas. The simultaneous occurrence of a functional adrenocortical adenoma and pheochromocytoma in the same gland is rare [2,3]. A few patients present with symptoms or biochemical laboratory findings compatible with pheochromocytoma and had both a pheochromocytoma and nonfunctioning adrenal cortical adenoma or subclinical Cushing syndrome [2]. Here, we report an extremely rare case of multiple adrenal incidentalomas comprising a functioning adrenal cortical adenoma with resulting adrenal Cushing syndrome and a pheochromocytoma, which was not detected by laboratory findings. A 58-year-old female with hypertension was hospitalized for evaluation and treatment of acute pneumonia. She complained of respiratory symptoms, edema, easy bruising, central obesity, weight gain, and proximal muscle weakness. Her blood examination showed hypokalemia with K of 2.9 mmol/L. She was taking furosemide, spironolactone, telmisartan, and amlodipine to correct the hypokalemia and control the edematous body changes. Chest computed tomography (CT) taken to evaluate the pneumonia incidentally revealed two discrete masses in the right adrenal gland. After 2 to 3 weeks of conservative treatment of the pneumonia, the patient was referred to our hospital for evaluation of the adrenal incidentalomas. On physical examination, she had a moon face and central obesity without abdominal cutaneous striae. Her family and social histories were unremarkable. Blood and urine samples were taken to evaluate the functional status of the tumor. She continued to take her antihypertensive medication and diuretics during the tests. The results revealed a normal serum aldosterone, metanephrine, normetanephrine, and 24-hour urinary catecholamines and metanephrines. Cushing syndrome studies showed abnormal values with an elevated serum cortisol (27.7 µg/dL) and 24-hour urinary cortisol (319 µg/day) (Table 1). Table 1 Hormone assays used to screen for adrenal adenoma An overnight dexamethasone suppression test was conducted to assess cortical secretion to screen for endogenous Cushing syndrome. The resulting cortisol level (22.6 µg/dL) was not suppressed by the overnight dexamethasone test. A standard 2-day dexamethasone suppression test with 0.5 mg per oral every 6 hours for 2 days was conducted for a definitive diagnosis of the Cushing syndrome. The serum cortisol and 24-hour urine cortisol were not suppressed by dexamethasone, remaining greater than 50 nmol/L, which confirmed the diagnosis of Cushing syndrome. CT of the adrenal glands showed two enhancing nodular lesions in the right adrenal gland that demonstrated rapid washout of contrast media consistent with cortical adenomas (Fig. 1). The patient underwent a laparoscopic right adrenalectomy, and two tumors were identified in the resected gland, as suggested by the CT imaging. The gross findings of the surgical specimens were compatible with adrenal cortical adenomas. The cut section shows two well-defined masses. The larger one, measuring 2.7 × 2.5 × 1.9 cm, was yellow and brown-tan with hemorrhage (Fig. 2A). The other, measuring 0.7 × 0.7 cm, was rubbery and yellow-white (Fig. 2B). Figure 1 Adrenal computed tomography findings. (A) The mass located at the lateral rim of the adrenal gland was 2.8 cm and its attenuation value was 27.7 Hounsfield unit (HU) initially, 158.5 HU at 1 minute, and 69.8 HU at 15 minutes. (B) The other mass measured ... Figure 2 Fresh right adrenalectomy surgical specimen, measuring 6.5 × 4.3 × 2.3 cm and weighing 22 g. The cut section shows two well-defined masses. Pathologically, the larger mass was diagnosed as an adrenal cortical adenoma and the smaller as ... The patient was stable intraoperatively and postoperatively. She was discharged from the hospital on prednisolone 10 mg per day. After discharge, the pathology report revealed two solid, well-encapsulated tumors with negative resection margins measuring 2.7 × 2.5 × 1.9 cm (Fig. 2A) and 1.0 × 1.0 cm (Fig. 2B). Microscopy and immunohistochemistry confirmed that the larger mass was a cortical adenoma that likely caused the Cushing syndrome (Fig. 2C), while the smaller mass was a pheochromocytoma (Fig. 2D). The immunohistochemical staining for chromogranin A was positive (×200) (Fig. 2E). The postoperative 24-hour urine cortisol was within the normal range (17 µg/day). A mixed corticomedullary tumor (MCMT) is a rare tumor, defined as a single adrenal lesion composed of a mixture of different cell types derived from both pheochromocytes and cortical cells, which do not share a common embryological origin [4]. Since MCMT was first described in 1969, at least 16 cases have been reported in the English- and French-language literature and more than 30 cases have been reported in three Japanese and two Chinese case series [4]. Coincident adrenal cortical adenomas and pheochromocytomas is not considered a category of MCMT and is extremely rare, especially in the same adrenal gland. There have been only 14 reported cases of a coexisting adrenal cortical adenoma and pheochromocytoma [4]. In all of these cases, there were no typical pheochromocytoma symptoms, and the disease was diagnosed by biochemical laboratory results and imaging studies [1]. Our patient presented with typical Cushingoid features and compatible CT imaging suggestive of an adrenal cortical adenoma. The plasma metanephrine and normetanephrine and the 24-hour urine metanephrine, normetanephrine, and catecholamines were normal, and there was no hypertensive crisis during the surgery. The final pathology examination revealed one mass consistent with an adrenocortical adenoma and a second 1-cm mass consistent with pheochromocytoma. Many pheochromocytomas are diagnosed incidentally during routine radiological examinations or rarely by genetic screening tests for multiple endocrine neoplasia (MEN) syndrome or von Hippel-Lindau disease [5]. There have been no incidental discoveries of surgically proven pheochromocytoma, except at autopsy [5]. Such unusual cases should lead us to evaluate the pheochromocytoma further, such as for gene mutations and MEN, if the patient is young. Roughly 10% of adrenal incidentalomas are bilateral or multiple masses. As in this case, multiple masses in the same adrenal gland that secrete different hormones are possible. Although there are no abnormal hormone tests for pheochromocytoma because the adrenal mass is too small to produce metanephrine, the coexistence of a different type of adrenal adenoma must be considered in all cases. We always test all screening hormone evaluations for adrenal incidentalomas. In conclusion, we reported the first case of two different adrenal masses, initially diagnosed as functioning adrenal adenomas, but eventually confirmed to be one cortisol-secreting adrenal adenoma and one histopathologically proven pheochromocytoma.

Male germ cell development includes mitotic and meiotic cell divisions that are followed by dramatic morphological changes resulting in the production of spermatozoa. Genetic evidence has indicated that the DAZ family genes are critical for successful male germ cell development in diverse animals as well as humans. In the present study, we investigated the cellular functions of Dazl in the mouse male germ cells. We identified a specific interaction of Dazl with the dynein light chain, a component of the dynein–dynactin motor complex. The subcellular distribution of Dazl was microtubule-dependent and a selected number of Dazl-bound mRNAs could accumulate in the perinuclear area. Based on these results, we propose that Dazl may play a role in transport of specific mRNAs via dynein motor complex. The Dazl-bound mRNAs may be stored at specific sites and would be available for future developmental processes. Our study revealed the presence of an active mRNA transport system in mouse male germ cells.

In this study, we investigated the effects of long-term administration of quercetin with or without daidzein on glutathione and the enzymes involved in its metabolism in rat liver in vivo. Male Sprague-Dawley rats were divided randomly into four groups and given oral quercetin (20 mg/day) and daidzein (20 mg/day) alone or in combination, or vehicle alone for six weeks. The serum and liver alpha-tocopherol concentrations were significantly increased following administration of quercetin and daidzein alone or in combination. Glutathione concentration and glutathione reductase activity was significantly (p0.05) decreased with quercetin treatment, while no such effect was observed with daidzein treatment. Interestingly, decrease in glutathione concentration and glutathione reductase activity by quercetin treatment was inhibited by combined administration of daidzein and quercetin. The malondialdehyde concentration was significantly decreased following administration of quercetin and daidzein alone or in combination. These results suggest that quercetin, but not daidzein, acts as a pro-oxidant agent by decreasing glutathione concentration and glutathione reductase activity. Interestingly, this pro-oxidant effect of quercetin was inhibited by the combined administration of quercetin and daidzein.

PURPOSE The purpose of this study was to investigate topographic characteristics of the retina in myopic eyes. METHODS We reviewed spectral-domain optical coherence tomography (SD-OCT) images of patients with myopia retrospectively. Retinal topography (RT) was defined as the topography of the retinal pigment epithelium layers. Retinal topographies were classified into several types, and the distribution and characteristics of each type were assessed in eyes with varying degrees of axial length (AL). RESULTS A total of 167 subjects with myopia were included in this study. Seventy eyes (41.9%) were classified as regular (R) type, 48 (28.7%) were wave (W) type, 32 (19.2%) were band (B) type, and 17 (10.2%) were pond (P) type. The distribution of these types varied significantly among eyes with different degrees of AL (P < 0.001); R-type was predominant in eyes with 24 mm less than or equal to AL less than 26 mm, and P-type in eyes with AL greater than or equal to 28 mm. The retina sloped away from the cornea more frequently on the nasal side than it did on the temporal side or in between the nasal and the temporal sides, with the exception of the P-type RT. Topography of the central zone was frequently slanted, except in B-type eyes. The difference between the refractive and corneal astigmatism of eyes with P-type RT was greater than that of B-type eyes (P = 0.002). CONCLUSIONS Retinal topography varies according to the degree of AL. This variation in RT may originate from the differences between healthy eyes and highly myopic eyes, and may be related to the optical characteristics of the eye.

The ER chaperone GRP78/BiP is crucial for the development of rheumatoid arthritis., An accumulation of misfolded proteins can trigger a cellular survival response in the endoplasmic reticulum (ER). In this study, we found that ER stress–associated gene signatures were highly expressed in rheumatoid arthritis (RA) synoviums and synovial cells. Proinflammatory cytokines, such as TNF and IL-1β, increased the expression of GRP78/BiP, a representative ER chaperone, in RA synoviocytes. RA synoviocytes expressed higher levels of GRP78 than osteoarthritis (OA) synoviocytes when stimulated by thapsigargin or proinflammatory cytokines. Down-regulation of Grp78 transcripts increased the apoptosis of RA synoviocytes while abolishing TNF- or TGF-β–induced synoviocyte proliferation and cyclin D1 up-regulation. Conversely, overexpression of the Grp78 gene prevented synoviocyte apoptosis. Moreover, Grp78 small interfering RNA inhibited VEGF165-induced angiogenesis in vitro and also significantly impeded synoviocyte proliferation and angiogenesis in Matrigel implants engrafted into immunodeficient mice. Additionally, repeated intraarticular injections of BiP-inducible factor X, a selective GRP78 inducer, increased synoviocyte proliferation and angiogenesis in the joints of mice with experimental OA. In contrast, mice with Grp78 haploinsufficiency exhibited the suppression of experimentally induced arthritis and developed a limited degree of synovial proliferation and angiogenesis. In summary, this study shows that the ER chaperone GRP78 is crucial for synoviocyte proliferation and angiogenesis, the pathological hallmark of RA.

Deletions of USP9Y have been observed among infertile males with defective spermatogenesis. Therefore, the gene has been designated as a male infertility gene on the Y chromosome. However, it remains to be determined how male infertility results from deletions of this gene. In order to initiate an investigation into the cellular functions of USP9Y in male germ cell development, in the present study we characterized the enzymatic specificity of USP9Y. Our results show that both USP9Y and Fam, the mouse infertility protein Usp9x, possess a protease activity specific to ubiquitin. These results suggest that, through de-ubiquitination, USP9Y may stabilize a specific target protein that is important for male germ cell development.