Pathological analysis of spermatic dysfunction following testicular ischemia-reperfusion injury\n

　Introduction & Objectives: Torsion, which may result in testicular ischemia, requires emergency surgery to restore testicular blood flow. However, the risk of spermatic dysfunction remains even if surgery is performed. The pathology of spermatic dysfunction in testicular ischemia-reperfusion injury (TIRI) remains unclear. A previous study showed the relevance of inflammation and oxidative stress in the other organs of ischemia-reperfusion injury. We hypothesized that inflammation and oxidative stress play key roles in causing spermatic dysfunction following TIRI. We investigated the pathophysiology of spermatic dysfunction in TIRI focusing on inflammatory changes using TIRI model mice.　Materials and Methods: The study used C57BL/6J male mice aged 10 to 15 weeks. To create TIRI model mice, the unilateral (left side) testicular vessels were clamped using Dieffenbach clamps (Bulldog clamps) for 1 hour and de-clamped. The bilateral testes were removed at 0 (ischemic state), 1, 3, and 5 weeks after creating the TIRI model mice. Spermatic changes following TIRI were investigated by analyzing the histology of the testes and semen and assessing levels of inflammation and oxidative stress. Semen was collected from the bilateral cauda epididymites and investigated using the sperm motility analysis system (SMAS).　Results: Histological analysis after hematoxylin-eosin staining showed tissue thickening in interstitial tissues at week 1 and 3 on the left (affected) testis, and week 1, 3 and 5 on the right (unaffected) testis. The infiltration of lymphocytes-predominant inflammatory cells were observed at week 1 and week 3 on the left (affected) testis. The destruction of ductal structures and giant cells were observed at weeks 3 and 5 on the left (affected) testis and week 5 on the right (unaffected) testis. SMAS showed significantly decreased spermatic concentration and motility in both testes of TIRI model mice compared with those of sham-operated mice at weeks 1, 3 and 5. Inflammation analysis using an inflammation-related proteome assay showed significantly increased levels of cytokines (IL-2, IL-3, IL-17A, and IL-23) and chemokines (CCL2, CCL5, CXCL1, and CX3CL1) at weeks 1, 3, and 5 in both testes of TIRI model mice. For the assessment of oxidative stress, enzyme-linked immuno-sorbent assay (ELISA) for 8-hydroxy-2’-deoxyguanosine (8-OHdG) was performed, which showed that levels of 8-OHdG were significantly increased in the left (affected) testis of TIRI model mice compared with that of sham-operated mice at all observation periods. Meanwhile, ELISA showed that levels of 8-OHdG in the right (unaffected) testis were significantly increased in TIRI model mice at weeks 3 and 5 compared with that of sham-operated mice.　Conclusions: Spermatic dysfunction following TIRI is induced by inflammation and oxidative stress. Inflammation and oxidative stress may be novel regulatory factors to prevent spermatic dysfunction following TIRI

Preventive effect of indoleamine 2,3-dioxygenase 1 inhibition on lipopolysaccharide-induced prostatitis

　Introduction and Objectives: Bacterial infections are the main cause of acute prostatitis and are treated with appropriate antimicrobial therapy. However, approximately 5% of patients continue to have inflammatory symptoms even after receiving antibacterial therapy, leading to refractory conditions. Bacterial prostatitis requires additional therapy, focusing on inflammatory changes. Indoleamine 2,3-dioxygenase 1 (IDO1) catalysis is the first rate-limiting step of tryptophan metabolism. IDO1 is expressed in the prostate and plays a key role in the immune response. As the first step in investigating the relationship between acute prostatitis and IDO1, we investigated the preventive effect of IDO1 inhibition on lipopolysaccharide (LPS)- induced prostatitis using IDO knockout (Ido1 −/−) mice in this study.　Materials and Methods: The study used Ido1 −/− and wild-type (Ido1 +/+) C57BL/6J malemice aged 10–15 weeks. LPS Escherichia coli O26 (100μg/PBS, 100μL) was administered transurethrally into the lower urinary tract to create a mouse model of LPS-induced prostatitis. The prostates were removed 1, 3, 5, and 7 days after creating the model mice. Histological, immunohistochemical, and biochemical analyses were used to compare the preventive effect in Ido1 −/− mice compared with that in Ido1+/+ mice.　Results: HE staining showed suppression of ductal destruction following infiltration of inflammatory cells in Ido1 −/− mice compared with Ido1 +/+ mice. The enzyme-linked immunosorbent assay (ELISA) method was used for kynurenine pathway analysis, which showed significantly maintained tryptophan levels and decreased L-kynurenine levels in Ido1 −/− mice compared to Ido1 +/+ mice. The IDO1 assay in Ido1 +/+ mice showed significantly increased levels during all observation periods after creating the model compared with that under normal conditions. Immunofluorescent staining using five types of cytokines and chemokines (IL-2, IL-4, IL-17, CCL2, and CCL3) related to the pathophysiology of acute prostatitis showed decreased expression of these cytokines and chemokines in Ido1 -/- mice compared with Ido1 +/+ mice. Inflammation-related proteome assays showed decreased levels of IL-1β, IL-4, IL-5, IL-6, IL-17, CCL2, CCL3, CXCL1, CXCL11, and tissue inhibitor of matrix metalloproteinases (TIMP)-1 in Ido1 −/− mice compared with Ido1 −/− mice during all observation periods after model creation.　Conclusions: IDO1 is involved in LPS-induced prostatitis through cytokines and chemokines. IDO1 inhibition contributes to the prevention of LPS-induced prostatitis. IDO1 inhibition has the potential to serve as an additional therapy for acute prostatitis

Pathophysiological analysis of detrusor overactivity following partial bladder outlet obstruction

　Introduction: Detrusor overactivity (DO) following partial bladder outlet obstruction (PBOO) is a common urological condition in humans, with 50-70% patients with PBOO complicated with DO. The pathological mechanisms of DO following PBOO are largely unknown, but inflammatory changes may play a key role. We hypothesized that inflammation is important in the earlier pathophysiological phase before overproduction of oxidative stress in DO following PBOO. Therefore, we investigated the relationships among bladder function, ischemia, oxidative stress and inflammation in DO following PBOO in PBOO model mice.　Materials and Methods: C57BL/6J male mice aged 10 to 15 weeks were used in the study. PBOO model mice were created surgically by ligation of the proximal urethra with 5-0 nylon suture under inhalation anesthesia. Sham-operated mice were used as controls. Pathophysiological changes in the bladder at 1, 3 and 5 weeks after creation of the PBOO model mice were compared with those in sham-operated mice using functional, histological, biochemical and immunohistochemical analyses.　Results: Functional analysis using a pressure flow study showed increased maximum detrusor pressure at 1 week and DO from 3 to 5 weeks after creation of the PBOO model. Histological analysis using hematoxylin-eosin and Masson-Trichrome staining showed greater invasion of inflammatory cells and fibrosis in PBOO model mice compared with sham-operated mice at 3 and 5 weeks. Inflammatory cells were mainly present in interstitial tissue, and fibrosis gradually infiltrated from interstitial tissue to the muscular layer. Ischemia analysis showed significantlyincreased HIF-1α in PBOO model mice at all time points. Oxidative stress analysis indicated significantly increased levels of ROS from 1 week and 8-OHdG from 3weeks in PBOO model mice. An inflammation-related proteome assay showed high levels of colony stimulating factor (CSF) family proteins at 1 week and IL-2, IL-3, IL-17A, IL-23, MMP-3, MMP-9 and periostin from 3 to 5 weeks in PBOO model mice.　Conclusions: Oxidative stress and inflammatory changes showed contemporaneous increase in pathophysiology of detrusor overactivity following partial bladder outlet obstruction. Especially, CSF family and ROS changes are showed in the early stage, and might be a predict marker in the pathophysiology of DO following PBOO at the early stage

Changes in serum antibody titers after vaccination for COVID-19 and evaluation of post-vaccination health conditions

　Introduction: The coronavirus disease 2019 (COVID-19) vaccine has preventive effects and high immunogenicity, but the outcomes of vaccination have not been widely reported. The goal of this study was to examine serum antibody titers before and after vaccination and to evaluate post-vaccination health conditions.　Methods: The subjects were 2,304 medical workers (mean age 41 years) at Kawasaki Gakuen who agreed to participate in the study and underwent COVID-19 vaccination, beginning in March 2021. Serum IgG antibody titers for SARS-CoV-2 spike protein were measured before the first vaccination and 4 weeks after the second vaccination. Health conditions were observed for 4 weeks after the second vaccination.　Results: The rates of seroconversion, seroprotection, and change in geometric mean antibody titer at 4 weeks after the second vaccination were 99.9%, 99.9%, and 2,685.5 (95% CI 587.8-5,319.2), respectively, suggesting high immunogenicity. After the first vaccination, pain, enlargement, and reddening occurred at the local injection site, and systemic side effects included fatigue, headache, physical pain, chill, nausea, and fever. After the second vaccination, the incidence of pain decreased, but those of other events increased. There were no serious side effects requiring hospitalization. In logistic regression analysis, sex, age, fever,chill, and lymph node enlargement after the second vaccination were associated with a change in antibody titer.　Conclusions: Serum antibody titers suggested high immunogenicity of the COVID-19 vaccine and a health condition survey confirmed the safety of the vaccine. Systemic side effects may serve as an index of immunization (acquisition of antibody) by the vaccine

Pathological analysis of spermatic dysfunction following testicular ischemia-reperfusion injury\n

Regular Articlejournal articl

Preventive effect of indoleamine 2,3-dioxygenase 1 inhibition on lipopolysaccharide-induced prostatitis

Regular Articlejournal articl

Pathophysiological analysis of detrusor overactivity following partial bladder outlet obstruction

Regular Articlejournal articl

Anti-inflammatory effect of tadalafil, a phosphodiesterase 5 (PDE5) inhibitor, in autoimmune prostatitis

　Prostatitis is one of the most common diseases in urology. The pathology involves intricate interactions among multiple factors, and this has limited the establishment of evidence-based treatment for prostatitis. An autoimmune prostatitis mouse model has recently been suggested to be appropriate as a model of human prostatitis pathology. PDE5 inhibitors are indicated as therapy for benign prostatic hyperplasia with lower urinary tract symptoms, pulmonary arterial hypertension and erectile dysfunction, and have also been shown to have an anti-inflammatory effect against tissue inflammation. Thus, we hypothesized that a PDE5 inhibitor may have a preventive effect on prostatitis. To verify this hypothesis, the anti-inflammatory effects of a PDE5 inhibitor, tadalafil, were examined in an autoimmune prostatitis mouse model. C57BL/6 male mice aged 15 weeks were used in the study. Prostate glands from Wistar rats aged 5 weeks were used as a source of prostate antigen (protein concentration 10 mg/ml). 　Prostate antigen (100 μg in 100 μl of adjuvant) was subcutaneously injected into C57BL/6 mice. Tadalafil (25 μg in 50 μl of water) was orally administered every day for up to 10 weeks after establishment of the model. Control mice received water only (50 μl). Inflammatory changes were investigated using histological, biochemical and immunohistochemical analyses. 　Histological analysis using hematoxylin-eosin and Masson-Trichrome staining showed inhibitory effects on tissue fibrosis following invasion by inflammatory cells in tadalafil-treated mice compared with control mice. Biochemical and immunohistochemical analyses using an inflammation-related proteome assay and immunostaining showed decreased levels of M-CSF, TREM-1, TIMP-1, CCL2, CCL3 and CXCL2 in tadalafil-treated mice compared with control mice.Tadalafil has an inhibitory effect on tissue fibrosis and decreases cytokine levels after an inflammatory response. These results suggest that PDE5 inhibitors might be effective as therapy for prostatitis