Evolutionary insights into strategy shifts for the safe and effective accumulation of ascorbate in plants.

Plants accumulate high concentrations of ascorbate, commonly in their leaves, as a redox buffer. While ascorbate levels have increased during plant evolution, the mechanisms behind this phenomenon are unclear. Moreover, has the increase in ascorbate concentration been achieved without imposing any detrimental effects on the plants? In this review, we focus on potential transitions in two regulatory mechanisms related to ascorbate biosynthesis and the availability of cellular dehydroascorbate (DHA) during plant evolution. The first transition might be that the trigger for the transcriptional induction of VTC2, which encodes the rate-limiting enzyme in ascorbate biosynthesis, has shifted from oxidative stress (in green algae) to light/photosynthesis (in land plants), probably enabling the continuous accumulation of ascorbate under illumination. This could serve as a preventive system against the unpredictable occurrence of oxidative stress. The second transition might be that DHA-degrading enzymes, which protect cells from the highly reactive DHA in green algae and mosses, have been lost in ferns or flowering plants. Instead, flowering plants may have increased glutathione concentrations to reinforce the DHA reduction capacity, possibly allowing ascorbate accumulation and avoiding the toxicity of DHA. These potential transitions may have contributed to strategies for plants' safe and effective accumulation of ascorbate.

Evaluating the State of Glomerular Disease by Analyzing Urinary Sediments: mRNA Levels and Immunofluorescence Staining for Various Markers.

Renal biopsy is the gold standard for making the final diagnosis and for predicting the progression of renal disease, but monitoring disease status by performing biopsies repeatedly is impossible because it is an invasive procedure. Urine tests are non-invasive and may reflect the general condition of the whole kidney better than renal biopsy results. We therefore investigated the diagnostic value of extensive urinary sediment analysis by immunofluorescence staining for markers expressed on kidney-derived cells (cytokeratin: marker for tubular epithelial cells, synaptopodin: marker for podocytes, claudin1: marker for parietal epithelial cells, CD68: marker for macrophages (MΦ), neutrophil elastase: marker for neutrophils). We further examined the expression levels of the mRNAs for these markers by real-time reverse transcription polymerase chain reaction. We also examined the levels of mRNAs associated with the M1 (iNOS, IL-6) and M2 (CD163, CD204, CD206, IL-10) MΦ phenotypes. Evaluated markers were compared with clinical and histological findings for the assessment of renal diseases. Claudin1- and CD68-positive cell counts in urinary sediments were higher in patients with glomerular crescents (especially cellular crescents) than in patients without crescents. The relative levels of mRNA for CD68 and the M2 MΦ markers (CD163, CD204, CD206, and IL-10) in urinary sediments were also higher in patients with glomerular crescents. These data suggest that immunofluorescence staining for claudin1 and CD68 in urinary sediments and the relative levels of mRNA for CD68 and M2 MΦ markers in urinary sediments are useful for evaluating the state of glomerular diseases.

Decreased plasma acetaminophen glucuronide/acetaminophen concentration ratio warns the onset of acetaminophen-induced liver injury.

Acetaminophen (APAP)-induced liver injury (AILI) is the most common cause of acute liver failure. Although the mechanisms that trigger AILI are well known, it is less understood how to halt AILI progression and facilitate liver recovery. Therefore, it is necessary to understand the pathophysiology of APAP hepatotoxicity in patients and to examine predictive/preventive markers. In a clinical study, we had a case in which aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels increased in a patient with a low ratio of APAP glucuronide concentration (AP-G)/APAP plasma concentration. Then a reverse translational study was conducted for clarifying this clinical question. The relationship between plasma AP-G/APAP concentration ratio and the levels of AST and ALT was examined by in vivo and in vitro experiments. In in vivo experiments, 10-week-old rats showed lower UGT activity, lower AP-G/APAP concentration ratios, and higher AST and ALT levels than 5-week-old rats. This suggests an inverse correlation between the AP-G/APAP concentration ratio and the AST, ALT levels in APAP-treated rats. Furthermore, as a result of the in vitro experiment, it was confirmed that the cell viability decreased when the AP-G/APAP concentration ratio in the culture medium decreased. Since the decrease in the plasma AP-G/APAP concentration ratio appears earlier than the increase of AST and ALT levels, the ratio might be a presymptomatic marker of AILI. When APAP is used for a long time, it is recommended to perform therapeutic drug monitoring of the AP-G/APAP concentration ratio, which is a predictive/preventive marker of AILI.

Transition from endocapillary proliferative glomerulonephritis to membranoproliferative glomerulonephritis in a patient with a prolonged human parvovirus B19 infection.

We report a case in which renal biopsies were performed 4 years apart in a woman with a prolonged human parvovirus B19 (HPB19) infection. When she was 29 years old the first biopsy, performed because of microscopic hematuria and mild proteinuria, showed endocapillary and mesangial proliferative glomerulonephritis in light microscopy as well as deposits of immunoglobulins (Igs) and complement C3 on capillary walls. Mesangial, intramembranous, and subepithelial hump-like electron dense deposits were seen in electron microscopy. The principal differential diagnoses, acute poststreptococcal glomerulonephritis and lupus nephritis, were unlikely, and her serological positivity for IgM antibody for HPB19 made us diagnose acute glomerulonephritis associated with HPB19 infection. The second biopsy, performed 4 years later because of persistent proteinuria and prolonged positivity for IgM antibody for HPB19, showed membranoproliferative glomerulonephritis (MPGN) with mesangial interposition and with thickening and double contours of glomerular basement membrane. In tissues obtained in both biopsies, HPB19 DNA was detected by polymerase chain reaction. HPB19 infection has been widely known to cause various glomerular diseases. This case reveals that acute endocapillary proliferative glomerulonephritis can change into MPGN during prolonged HPB19 infection.

[A Questionnaire Survey of Nurses on Pharmacist Services in the Emergency Rooms at Tokushukai Medical Group].

We conducted a multicenter survey of emergency room nurses to obtain information that would be useful for the establishment of pharmacist services in emergency rooms. Notably, 199 valid responses were obtained from 12 hospitals. The most common expectation from pharmacists in the emergency room was "drug management" (70.9%), followed by "providing information to physicians regarding the patient's medication history" (59.3%), and "auditing of dosage and interaction" (57.3%). The working arrangements that the survey respondents wanted regarding pharmacists in emergency rooms were: 24 h pharmacist (41.7% wanted this arrangement), day-shift pharmacist (24.6% wanted this arrangement), 24 h on-call (17.1% wanted this arrangement), day-shift on-call (5.0% wanted this arrangement), telephone support (11.1% wanted this arrangement), and 0.5% said that there was no need for pharmacists. In the analysis of factors affecting nurse satisfaction, day-shift pharmacist was a significant factor. We hope that the results of this survey will be used as a guide for the development of emergency room pharmacist services tailored to the unique characteristics and actual working conditions of each hospital.

Protective effects of Rho kinase inhibitor fasudil on rats with chronic kidney disease.

The protective effects of Rho kinase inhibitor fasudil against renal diseases have recently been reported. We compared the therapeutic effects of fasudil on the spontaneously hypercholesterolemic (SHC) rat, a model of chronic kidney disease (CKD) with proteinuria, with those of the angiotensin receptor blocker olmesartan (OL) by paying attention to the proteinuria and the macrophage phenotype. SHC rats were allocated to six treatment groups: a vehicle (Ve) group, a low-dose fasudil (FL) group, a high-dose fasudil (FH) group, an OL group, a combination of low-dose fasudil and OL (CL) group, and a combination of high-dose fasudil and OL (CH) group. Sprague-Dawley rats treated with vehicle served as a control (n = 7/each). The rats were treated for 24 wk. Compared with the Ve group, proteinuria was significantly decreased in the FH, OL, and CL groups, and it completely disappeared in the CH group. Glomerular stainings of nephrin and F-actin were focally impaired in the Ve group but were restored in the CH group. Western blotting showed that the CH group had significantly increased renal nephrin expression compared with the Ve group. Interstitial infiltration of macrophages was significantly increased in the Ve group, which was significantly attenuated in all treatment groups. The ratio of CD206 (M2 macrophage marker) to CD68 mRNA was significantly greater in the CH group than in the Ve group. These results indicate that fasudil with OL reduces proteinuria by protecting podocyte integrity and alters the interstitial macrophage density/phenotype, thereby exerting renoprotective effects against CKD.

Clinical and immunological implications of increase in CD208+ dendritic cells in tonsils of patients with immunoglobulin A nephropathy.

BACKGROUND: The therapeutic effect of tonsillectomy for immunoglobulin A nephropathy (IgAN) has been widely recognized, but the mechanism by which tonsillar immunity leads to glomerulonephritis has been unclear. We investigated subtypes and localization of dendritic cells (DCs) in tonsils and looked for relationships between the tonsillar DCs and the clinical features and renal histological changes of patients with IgAN. METHODS: We examined tonsils from 33 IgAN patients, using as control tonsillar specimens from subjects without glomerulonephritis. Five distinct markers of DCs (CD303, CD1c, CD209, CD208 and CD1a) were analyzed by immunohistochemistry and flow cytometry. The mRNA levels of these DC markers were evaluated using real-time polymerase chain reaction. The clinical data and histological results obtained evaluating renal biopsy tissues were statistically compared with immunological data. RESULTS: Of the five subtypes of DCs, CD208(+) DCs were significantly increased in the tonsils of IgAN patients compared with that of controls. Furthermore, the number of CD208(+) DCs in the tonsils was positively and linearly correlated with the proportion of crescentic glomeruli in renal biopsy tissues and with the urinary protein level. Only few CD208(+) cells, however, were found in the kidney biopsy specimens of IgAN patients. CONCLUSIONS: These observations suggest that increased CD208(+) DCs in tonsils may play a directive role in the pathogenesis of IgAN. The present results support the therapeutic significance of tonsillectomy for IgAN patients.

Renal tamponade in a patient with hydronephrosis-related Page kidney.

A 48-year-old woman presented with hyperreninemic hypertension and renal dysfunction and was diagnosed with hydronephrosis-related Page kidney. The pathophysiology was "renal tamponade", in which the kidney was compressed by the renal pelvis and Gerota's fascia, resulting in intrarenal microvascular ischemia. Ureteral stent placement promptly improved the hyperreninemic hypertension and renal dysfunction, and additional perirenal fluid drainage gradually improved these conditions. These observations indicated the following three points. First, renal compression-induced renin-angiotensin-aldosterone system upregulation plays an important role in the pathogenesis of Page kidney. Second, physicians should consider perirenal fluid drainage as a therapeutic option in addition to ureteral stenting in patients with hydronephrosis-related Page kidney. Third, bilateral perirenal subcapsular hematomas in this case could be caused by hydronephrosis. Hydronephrosis-induced intrarenal pressure elevation possibly caused chronic perirenal subcapsular hemorrhage at the vulnerable sites of the renal cortex and peeling of the renal capsule from the cortex, resulting in the bilateral massive subcapsular hematomas and Page kidney.

Identifying Clusters of Health Behaviors in a Japanese Working Population at Risk for Non-Communicable Diseases: A Latent Class Analysis of 12,168 Individuals.

Introduction: Noncommunicable diseases (NCDs) have become a significant global problem. Health behaviors are associated with NCDs, and characterizing populations using a public health approach can help provide specific interventions according to their characteristics. This study aims to examine the formation of clusters of health behavior combinations in the Japanese working population at risk of NCDs, taking into account the influences of age and gender, using latent class analysis. Methods: Participants were individuals at risk for NCDs but had not previously been diagnosed with any. Latent class analysis (LCA) was used to study clustering based on basic characteristics and health behaviors. All statistical analyses were conducted using R (Version 4.0.4) and the "poLCA" package (Version 1.6.0). Results: This study included 12,168 participants. LCA compared models with one to six latent classes. The five-class model was determined to be the most appropriate based on Bayesian Information Criterion, Akaike Information Criterion, and G^2 values, as well as distinguishable cluster characteristics. Cluster 1: "having healthy lifestyles but disliking hospitals"; Cluster 2: "women with healthy lifestyle behaviors"; Cluster 3: "general population"; Cluster 4: "middle-aged group in need of lifestyle improvement"; Cluster 5: "a group receiving treatment for lifestyle-related diseases." Conclusions: This study reveals discernible health behavior patterns in a sample of the Japanese population using large real-world data, suggesting the effectiveness of distinct approaches when considering a population approach to public health.

The role of nephritis-associated plasmin receptor (NAPlr) in glomerulonephritis associated with streptococcal infection.

It is well known that glomerulonephritis can occur after streptococcal infection, which is classically referred to as acute poststreptococcal glomerulonephritis (APSGN). The pathogenic mechanism of APSGN has been described by so-called immune complex theory, which involves glomerular deposition of nephritogenic streptococcal antigen and subsequent formation of immune complexes in situ and/or the deposition of circulating antigen-antibody complexes. However, the exact entity of the causative antigen has remained a matter of debate. We isolated a nephritogenic antigen for APSGN from the cytoplasmic fractions of group A streptococcus (GAS) depending on the affinity for IgG of APSGN patients. The amino acid and the nucleotide sequences of the isolated protein revealed to be highly identical to those of reported plasmin(ogen) receptor of GAS. Thus, we termed this antigen nephritis-associated plasmin receptor (NAPlr). Immunofluorescence staining of the renal biopsy tissues with anti-NAPlr antibody revealed glomerular NAPlr deposition in essentially all patients with early-phase APSGN. Furthermore, glomerular plasmin activity was detected by in situ zymography in the distribution almost identical to NAPlr deposition in renal biopsy tissues of APSGN patients. These data suggest that NAPlr has a direct, nonimmunologic function as a plasmin receptor and may contribute to the pathogenesis of APSGN by maintaining plasmin activity.

Seroma as a Rare Complication of Autologous Arteriovenous Fistula Creation in the Forearm of a Hemodialysis Patient: A Case Report.

Aim of the study: Seromas are rarely reported as complications of autologous arteriovenous fistula creation. Case description: An 89-year-old woman was hospitalized for hemodialysis and underwent an autologous arteriovenous fistula creation in the forearm. During cephalic vein expansion using a heparinized saline solution, leakage occurred. A suture was placed to control the leakage, and a Penrose drain was inserted. Serosanguineous drainage ceased on postoperative day two; however, a seroma occurred approximately two weeks after the surgery. Follow-up ultrasonography revealed no growth tendency; therefore, excision and aspiration were unnecessary. Conclusion: This seroma was associated with postoperative dead space, surgical technique, and patient clinical status. Sufficient preoperative ultrasonographic vascular mapping is required to avoid inappropriate handling of veins and prevent seroma formation. Postoperative ultrasonographic follow-up is recommended due to the future risk of fistula dysfunction and infection associated with seroma enlargement, which may necessitate surgical seroma excision.

A Mobile Health-Based Disease Management Program Improves Blood Pressure in People With Multiple Lifestyle-Related Diseases at Risk of Developing Vascular Disease　- A Retrospective Observational Study.

Background: The overlap of multiple lifestyle-related diseases increases the risk of vascular diseases. This study investigated the effects of a mobile health (mHealth)-based disease management program on blood pressure and the safety of this program in people with multiple lifestyle-related diseases at risk of developing vascular disease. Methods and Results: This retrospective observational study was conducted using secondary data collected by PREVENT Inc. People with a full history of hypertension, diabetes, and dyslipidemia and who participated in a 6-month mHealth-based disease management program were included in the study. The primary outcome was blood pressure. Adverse events during the program were investigated to evaluate safety. In total, 125 participants (mean [±SD] age 55.3±6.2 years) were examined. Systolic and diastolic blood pressure were significantly lower after the intervention than at baseline (systolic blood pressure, 128.0±12.3 vs. 131.9±12.7 mmHg [P<0.001]; diastolic blood pressure, 81.2±9.3 vs. 83.6±8.9 mmHg; P=0.003). No serious adverse events occurred during the program. Conclusions: The present results indicate that the mHealth-based disease management program may reduce blood pressure in people with multiple lifestyle-related diseases at risk of developing vascular disease and that the program is safe. These findings will help shape future health instructions using mHealth-based interventions.

Putative Novel Species of Genus Capnocytophaga, Capnocytophaga stomatis Bacteremia in a Patient with Multiple Myeloma after Direct Contact with a Cat.

Capnocytophaga species are among the typical zoonotic pathogens causing infections following direct contact with animals. Recently, a putative novel species of zoonotic Capnocytophaga, Capnocytophaga stomatis, was reported. We herein report the first case of bacteremia caused by C. stomatis. A woman in her 80s with multiple myeloma who was receiving bortezomib and dexamethasone therapy was admitted to our hospital with a 2-day history of a fever and right calf redness. She was often licked by her cat. On a blood culture, thin, Gram-negative rods were detected, which were identified as C. stomatis by whole-genome sequencing. The patient was successfully treated with ampicillin-sulbactam treatment. Our case highlights the pathogenic potential of the putative novel Capnocytophaga, C. stomatis, in immunocompromised hosts.

Effects of liposome-encapsulated clodronate on chlorhexidine gluconate-induced peritoneal fibrosis in rats.

BACKGROUND: Long-term peritoneal dialysis (PD) causes morphologic and functional changes in the peritoneum that hamper the continuation of PD therapy. Because macrophages play important roles in the development of peritoneal fibrosis and liposome-encapsulated clodronate (LC) induces macrophage apoptosis, we examined the effect of LC on chlorhexidine gluconate (CG)-induced peritoneal fibrosis in rats. METHODS: Fifty Sprague-Dawley rats were randomly allocated into five groups of 10 receiving intraperitoneal (i.p.) injections (1.5 mL/100 g) of either 0.1% CG (four groups) or vehicle (one group) three times a week. Three of the CG-treated groups also received intravenous injections of clodronate twice a week: 10 mg of LC, 20 mg of LC or 20 mg of unencapsulated clodronate (UC20). Twenty-one days after the first i.p. injection, the rats were sacrificed and the parietal peritoneum was harvested. RESULTS: The number of peritoneal macrophages in the rats given clodronate was significantly smaller than that in rats not given clodronate (92.0 ± 4.6 cells per field). It was 54.1 ± 3.2 cells per field in the group given 20 mg UC, 43.2 ± 5.2 cells per field in the group given 10 mg LC and 27.2 ± 2.8 cells per field in the group given 20 mg LC. This decrease in macrophage number was paralleled by decreases in peritoneal thickening, in the number of mesothelial cells staining positive for cytokeratin and α-smooth muscle actin and in messenger RNA expression for transforming growth factor-ß1 and collagen types I and III. CONCLUSIONS: These data suggest that macrophages play a critical role in the development of peritoneal fibrosis and that LC may be useful for treating peritoneal fibrosis in PD patients.

Alteration in the phenotype of macrophages in the repair of renal interstitial fibrosis in mice.

AIM: Renal interstitial fibrosis is the final common pathway determining long-term prognosis of chronic kidney diseases, but its repair process is scarcely understood. Because recent reports indicate that M2 macrophages play important roles in the repair of various tissues, special attention was paid to the phenotypes of infiltrating macrophages in the present study when the histological changes occurring in mouse kidneys after the release of unilateral ureteral obstruction (UUO) inducing renal fibrosis were analyzed. METHODS: The left ureter of male mice was obstructed for 10 days by using a vascular clamp, and that kidney was removed for analysis either on the day when the clamp was removed or after the kidney had been allowed to recover for 3, 7 or 21 days. RESULTS: Interstitial fibrosis assessed by picrosirius red staining decreased with time after the release, and this decrease was paralleled by a decrease in the interstitial area positive for α-smooth muscle actin. Macrophage infiltration assessed by F4/80 staining also significantly decreased from day 3. In contrast, real-time reverse transcription polymerase chain reaction revealed that the ratios of mRNA for the macrophage scavenger receptor (CD204) and the mannose receptor (CD206), both of which are preferentially expressed on M2 macrophages, to CD68 (a general macrophage marker) were significantly greater on day 7 than on day 0 in the UUO-released mice. CONCLUSION: Although the total number of infiltrating myofibroblasts and macrophages decreased after UUO release, the ratios of macrophages expressing CD204 and CD206 increased, suggesting that M2 macrophages play an important role in the repair of renal fibrosis.

Effects of endoscopy-related procedure time on musculoskeletal disorders in Japanese endoscopists: a cross-sectional study.

Background and study aims There has been little evidence assessing the prevalence of musculoskeletal disorders (MSDs) among endoscopists performing recent diagnostic and therapeutic endoscopic procedures requiring prolonged procedural times. We evaluated the prevalence and identified the risk factors for developing MSDs, focusing on procedural time. Methods An electronic survey of endoscopists (n = 213) employed at the Nagoya University Hospital and its affiliated hospitals was developed by a multidisciplinary group.  Results Of the 110 endoscopists (51.6 %) who responded to the survey, eighty-seven endoscopists (79.1 %) had experienced endoscopy-related MSDs during the previous 1 year, and 49 endoscopists (44.5 %) had experienced these MSDs during the previous week. Nineteen endoscopists (17.3 %) reported absence from work due to severe MSDs. The most frequent sites of MSDs were neck, low back, and shoulders. Logistic regression analyses showed that longer upper endoscopic submucosal dissection ESD, (odds ratio: 5.7; 95 %CI: 1.3-25.0), lower ESD (odds ratio 4.9; 95 %CI: 1.1-22.0), and lower gastrointestinal treatment (odds ratio: 5.6; 95 %CI: 2.3-13.3) were significantly associated with the development of MSDs in the low back area. Moreover, longer lower ESD (odds ratio: 5.0; 95 % CI: 1.2-20.2) was a risk factor for symptoms in the left shoulder. Conclusion This study suggests a correlation between the volume of therapeutic endoscopic procedures including ESD and the risk of MSDs mainly low back area and left shoulder. Managing monthly total endoscopic time, in light of organizational ergonomics, could contribute to minimizing such risks of endoscopy-related MSDs.

Acute Brachial Arterial Embolic Occlusion Following Anticoagulant Discontinuation in a Renal Biopsy of a Nephrotic Syndrome Patient.

A 73-year-old woman with atrial fibrillation treated with rivaroxaban was hospitalized for nephrotic syndrome. After discontinuation of rivaroxaban to lower the risk of hemorrhagic events, a renal biopsy was performed. Rivaroxaban was scheduled to resume a week after the biopsy to prevent renal hemorrhaging. However, she developed acute brachial arterial embolic occlusion and mural thrombosis in the abdominal aorta before resuming rivaroxaban. If immune-mediated renal diseases are suspected in anticoagulated patients at a risk of thrombotic events, physicians should consider initiating glucocorticoid therapy without a renal biopsy in order to avoid hemorrhagic and thrombotic events.

Severe anaphylaxis caused by intravenous anti-cancer drugs.

BACKGROUND: The incidence and risk factors of severe anaphylaxis by intravenous anti-cancer drugs are unclear, whereas those of milder reactions have been reported. STUDY DESIGN: Electronic medical charts of cancer patients who have undergone intravenous chemotherapy between January 2013 and October 2020 in a university hospital were retrospectively reviewed. Non-epithelial malignancies were also included in the analysis. "Severe anaphylaxis" was judged using Brown's criteria: typical presentation of anaphylaxis and one or more of hypoxia, shock, and neurologic compromise. (UMIN000042887). RESULTS: Among 5584 patients (2964 males [53.1%], 2620 females [46.9%], median age 66 years), 88,200 person-day anti-cancer drug administrations were performed intravenously, and 27 severe anaphylaxes were observed. The causative drugs included carboplatin (14 cases), paclitaxel (9 cases), and cisplatin, docetaxel, trastuzumab, and cetuximab (1 case each). The person-based lifetime incidence of severe anaphylaxis for patients who received at least one intravenous chemotherapy was 0.48% (27/5584, 95% confidence interval (CI) 0.30%-0.67%) and the administration-based incidence was 0.031% (27/88,200, 95% CI 0.019%-0.043%). Among 124 patients who received at least 10 carboplatin administrations, 10 patients experienced carboplatin-induced severe anaphylaxis (10/124, 8.1%, 95% CI 3.0%-13.1%). Carboplatin caused severe anaphylaxis after at least 9-min interval since the drip started. Thirteen out of 14 patients experienced carboplatin-induced severe anaphylaxis within a 75-day interval from the previous treatment. Paclitaxel infusion caused severe anaphylaxis after a median of 5 min after the first drip of the day at a life-long incidence of 0.93% (9/968, 95% CI 0.27%-1.59%). CONCLUSION: We elucidated the high-risk settings of chemotherapy-induced severe anaphylaxis.

Voltage-gated potassium channel Kv1.3 blocker as a potential treatment for rat anti-glomerular basement membrane glomerulonephritis.

The voltage-gated potassium channel Kv1.3 has been recently identified as a molecular target that allows the selective pharmacological suppression of effector memory T cells (T(EM)) without affecting the function of naïve T cells (T(N)) and central memory T cells (T(CM)). We found that Kv1.3 was expressed on glomeruli and some tubules in rats with anti-glomerular basement membrane glomerulonephritis (anti-GBM GN). A flow cytometry analysis using kidney cells revealed that most of the CD4(+) T cells and some of the CD8(+) T cells had the T(EM) phenotype (CD45RC(-)CD62L(-)). Double immunofluorescence staining using mononuclear cell suspensions isolated from anti-GBM GN kidney showed that Kv1.3 was expressed on T cells and some macrophages. We therefore investigated whether the Kv1.3 blocker Psora-4 can be used to treat anti-GBM GN. Rats that had been given an injection of rabbit anti-rat GBM antibody were also injected with Psora-4 or the vehicle intraperitoneally. Rats given Psora-4 showed less proteinuria and fewer crescentic glomeruli than rats given the vehicle. These results suggest that T(EM) and some macrophages expressing Kv1.3 channels play a critical role in the pathogenesis of crescentic GN and that Psora-4 will be useful for the treatment of rapidly progressive glomerulonephritis.

Involvement of epimorphin in the repair of experimental renal fibrosis in mice.

Interaction between epithelial cells and mesenchymal cells is essential in normal organ morphogenesis and in tissue repair after injury. Epimorphin, a mesenchymal protein that regulates epithelial morphogenesis through epithelial-mesenchymal interactions, has recently attracted attention as an important modulator of tissue repair. In this study we analyzed the role of epimorphin in renal fibrosis. We first found a progressive increase in epimorphin expression corresponding to the progression of renal fibrosis in mice with unilateral ureteral obstruction (UUO). To determine whether this expression has a role in the repair or progression of renal fibrosis, we analyzed a model of renal fibrosis repair, the UUO-release (UUO-R) model. Epimorphin expression was increased at 3 and 7 days after the UUO-R rather than on the day of release, but was decreased at 21 days after the release. Inhibition of endogenous epimorphin with anti-epimorphin antibody (MC-1) significantly delayed the repair of fibrosis. When compared with normal-IgG-injected mice, MC-1-injected mice showed significantly decreased renal matrix metalloproteinase (MMP)-2 and MMP-9 expressions by western blotting and increased expression of TGF-beta and collagen-I mRNA by real-time RT-PCR. Recombinant epimorphin induced prominent increases in MMP-2 and MMP-9 activities in the culture media of renal interstitial fibroblasts in vitro. These findings indicate that epimorphin has a pivotal role in the repair of renal fibrosis by modulating both extracellular matrix (ECM) degradation and its production.