Switching mechanism from AR to EGFR signaling via 3-O-sulfated heparan sulfate in castration-resistant prostate cancer.

Androgen deprivation therapy is given to suppress prostate cancer growth; however, some cells continue to grow hormone-independently as castration-resistant prostate cancer (CRPC). Sulfated glycosaminoglycans promote ligand binding to receptors as co-receptors, but their role in CRPC remains unknown. Using the human prostate cancer cell line C4-2, which can proliferate in hormone-dependent and hormone-independent conditions, we found that epidermal growth factor (EGF)-activated EGFR-ERK1/2 signaling via 3-O-sulfated heparan sulfate (HS) produced by HS 3-O-sulfotransferase 1 (HS3ST1) is activated in C4-2 cells under hormone depletion. Knockdown of HS3ST1 in C4-2 cells suppressed hormone-independent growth, and inhibited both EGF binding to the cell surface and activation of EGFR-ERK1/2 signaling. Gefitinib, an EGFR inhibitor, significantly suppressed C4-2 cell proliferation and growth of a xenografted C4-2 tumor in castrated mouse. Collectively, our study has revealed a mechanism by which cancer cells switch to hormone-independent growth and identified the key regulator as 3-O-sulfated HS.

Effect of annealing on the magnetic microstructure of high-pressure torsion iron: the relevance of higher-order contributions to the magnetic small-angle neutron scattering cross section.

The development of higher-order micromagnetic small-angle neutron scattering theory in nanocrystalline materials is still in its infancy. One key challenge remaining in this field is understanding the role played by the microstructure on the magnitude and sign of the higher-order scattering contribution recently observed in nanocrystalline materials prepared by high-pressure torsion. By combining structural and magnetic characterization techniques, namely X-ray diffraction, electron backscattered diffraction and magnetometry with magnetic small-angle neutron scattering, this work discusses the relevance of higher-order terms in the magnetic small-angle neutron scattering cross section of pure iron prepared by high-pressure torsion associated with a post-annealing process. The structural analysis confirms: (i) the preparation of ultra-fine-grained pure iron with a crystallite size below 100 nm and (ii) rapid grain growth with increasing annealing temperature. The analysis of neutron data based on the micromagnetic small-angle neutron scattering theory extended to textured ferromagnets yields uniaxial magnetic anisotropy values that are larger than the magnetocrystalline value reported for bulk iron, supporting the existence of induced magnetoelastic anisotropy in the mechanically deformed samples. Furthermore, the neutron data analysis revealed unambiguously the presence of non-negligible higher-order scattering contributions in high-pressure torsion iron. Though the sign of the higher-order contribution might be related to the amplitude of the anisotropy inhomogeneities, its magnitude appears to be clearly correlated to the changes in the microstructure (density and/or shape of the defects) induced by combining high-pressure torsion and a post-annealing treatment.

A Case of Placenta Accreta Infection Managed by Hysterectomy After Treatment for Retained Products of Conception With Uterine Artery Embolization and Hysteroscopy.

Retained products of conception (RPOC) are frequently associated with previous cesarean section (C-section), abortion, and intrauterine operations, which may affect subsequent pregnancies. A 38-year-old female had a history of C-section and two abortions. After the second abortion, she underwent evacuation of RPOC and was treated with uterine artery embolization (UAE) and hysteroscopic resection. She became pregnant again and vaginally delivered an infant at full term. After delivery, RPOC was suspected on magnetic resonance imaging (MRI), but the patient was discharged for follow-up. She was rehospitalized with a diagnosis of infection and a placental remnant. Antibiotics did not improve the infection; therefore, she underwent a total hysterectomy. After the operation, signs of infection rapidly improved. The pathological diagnosis was placenta accreta. This case was considered a high-risk group for RPOC. In such rare and complicated cases, it is important to consider the possibility of recurrent RPOC and provide sufficient explanations before delivery for subsequent intensive management.

Ovarian Follicular Lymphoma Diagnosed Due to Hydronephrosis.

A 74-year-old woman presented with left lateral abdominal pain. Abdominal echography revealed left hydronephrosis and a pelvic mass. The patient underwent left adnexal resection of a suspected left ovarian tumor and was diagnosed with follicular lymphoma (FL) of clinical stage IIIA, grade 2. The patient was treated with rituximab-combined chemotherapy and achieved complete remission. The most common histological types of ovarian lymphoma are diffuse large B-cell lymphoma and Burkitt lymphoma, with FL being an extremely rare variant. We herein report a case of ovarian FL diagnosed as hydronephrosis.

Model experiments on hydraulic properties around multiple piers with reproduced 3D geometries.

The effects of multiple piers with 3D geometries on water levels during floods, as well as riverbed fluctuations and local scour after floods were examined via hydraulic model experiments. The Kintaikyo Bridge in Japan, a world-famous historical triple-wooden-arch bridge, was modeled at a scale of approximately 1/276. The number of model piers was set to four, as in the real bridge, and they were 3D printed and installed in the sand layer of an open channel. To provide adequate information on the pier installation conditions and hydraulic response, four cases were studied in the experiments, primarily considering present pier conditions; the previous conditions of the Kintaikyo Bridge were also considered. The experimental conditions in these four cases depended on whether foundations were present, the piers were skewed against the flow center, and the shape was spindle or similar to that of the present case. In the current bridge condition where foundations are present and there is no skew, water level rise, riverbed fluctuation, and local scour were suppressed. Furthermore, the statistical analysis of the results suggested that riverbed fluctuations are more stable after flooding. The two piers with foundations had smaller scour areas than those without. Further, those with foundations generally had lower mean scour at the pier perimeters. Regarding those without foundations, there were no differences in riverbed fluctuations or local scour due to the skewed pier angle. In addition, the representative length of the pier with a 3D geometry was determined by dividing the projected area of the pier below the initial sand surface in the main flow direction by the pier height, and the relationship between the Froude number or the mean water depth and the maximum scour depth was investigated.

Impact of Catheter Ablation on Brain Microstructure and Blood Flow Alterations for Cognitive Improvements in Patients with Atrial Fibrillation: A Pilot Longitudinal Study.

Atrial fibrillation (AF) predisposes patients to develop cognitive decline and dementia. Clinical and epidemiological data propose that catheter ablation may provide further benefit to improve neurocognitive function in patients with AF, but the underlying mechanism is poorly available. Here, we conducted a pilot prospective study to investigate whether AF ablation can alter regional cerebral blood flow (rCBF) and brain microstructures, using multimodal magnetic resonance imaging (MRI) technique. Eight patients (63 ± 7 years) with persistent AF underwent arterial-spin labeling (ASL) perfusion, 3D T1-structural images and cognitive test batteries before and 6 months after intervention. ASL and structural MR images were spatially normalized, and the rCBF and cortical thickness of different brain areas were compared between pre- and 6-month post-treatment. Cognitive-psychological function was improved, and rCBF was significantly increased in the left posterior cingulate cortex (PCC) (p = 0.013), whereas decreased cortical thickness was found in the left posterior insular cortex (p = 0.023). Given that the PCC is a strategic site in the limbic system, while the insular cortex is known to play an important part in the central autonomic nervous system, our findings extend the hypothesis that autonomic system alterations are an important mechanism explaining the positive effect of AF ablation on cognitive function.

Effects of obesity on CC16 and their potential role in overweight/obese asthma.

INTRODUCTION: Club cell secretory protein-16 (CC16) is a major anti-inflammatory protein expressed in the airway; however, the potential role of CC16 on overweight/obese asthma has not been assessed. In this study, we examined whether obesity reduces airway/circulatory CC16 levels using experimental and epidemiological studies. Then, we explored the mediatory role of CC16 in the relationship of overweight/obesity with clinical asthma measures. METHODS: Circulating CC16 levels were assessed by ELISA in three independent human populations, including two groups of healthy and general populations and asthma patients. The percentage of cells expressing club markers in obese vs. non-obese mice and human airways was determined by immunohistochemistry. A causal mediation analysis was conducted to determine whether circulatory CC16 acted as a mediator between overweight/obesity and clinical asthma measures. RESULTS: BMI was significantly and monotonously associated with reduced circulating CC16 levels in all populations. The percentage of CC16-expressing cells was reduced in the small airways of both mice and humans with obesity. Finally, mediation analysis revealed significant contributions of circulatory CC16 in the association between BMI and clinical asthma measures; 21.8% of its total effect in BMI's association with airway hyperresponsiveness of healthy subjects (p = 0.09), 26.4% with asthma severity (p = 0.030), and 23% with the required dose of inhaled corticosteroid (p = 0.042). In logistic regression analysis, 1-SD decrease in serum CC16 levels of asthma patients was associated with 87% increased odds for high dose ICS requirement (p < 0.001). CONCLUSIONS: We demonstrate that airway/circulating CC16, which is inversely associated with BMI, may mediate development and severity in overweight/obese asthma.

Severe Abdominal Recurrence of Low-grade Endometrial Stromal Sarcoma After Hysteroscopic Surgery.

BACKGROUND: Low-grade endometrial stromal sarcoma (LG-ESS) is an indolent tumor harboring gene fusion involving polycomb family genes. While early LG-ESS has a good clinical course, some tumors have pelvic recurrence. The etiology and genetic alterations involved in the process remain unknown. CASE REPORT: A 44-year-old nulliparous woman underwent hysteroscopic surgery for a 2.5 cm submucosal uterine tumor with negative endometrial cytology. Pathological evaluation revealed LG-ESS. On the 31st day, total laparoscopic hysterectomy was indicated. She was diagnosed with stage IA (pT1aNXM0) LG-ESS without lymphovascular invasion. At 4 years, positron-emission tomography showed multiple pelvic masses. Secondary debulking surgery was performed, which revealed severe intra-abdominal recurrence of LG-ESS with JAZF1-SUZ12 fusion. CONCLUSION: Hysteroscopic surgery is a convenient tool for benign uterine submucosal diseases. However, intrauterine morcellation with fluid can lead to unexpected recurrence of occult LG-ESS. It is important when seeking consent for surgery to inform patients about the possible risk of dissemination of uterine mesenchymal tumors.

A case report of advanced ovarian cancer detected due to an inguinal metastasis in the canal of Nuck.

INTRODUCTION: Ovarian cancer has few subjective symptoms, so approximately 40%-50% of cases have already reached stage III or IV by the time of diagnosis. These are advanced stages of the disease and have poor prognosis. Among these cases, less than 3 % are reported to exhibit inguinal lymph node metastasis. This report documents a rare case of advanced ovarian cancer that we detected due to an inguinal metastasis in the canal of Nuck. The work has been reported in line with the SCARE criteria. PRESENTATION OF CASE: The patient was a 43-year-old, married, premenopausal woman (G2P1). She was examined by her local practitioner for a chief complaint of a mass in the right inguinal region and was found to have a right inguinal mass. Magnetic resonance imaging (MRI) scans revealed a left ovarian tumor, and she was referred to our department. Rapid intraoperative diagnosis showed a highly atypical serous carcinoma present in both the left ovary and the right inguinal region mass, where the tumor had extended into the canal of Nuck. DISCUSSION: In this case, the right inguinal mass was ovarian cancer that had metastasized to a cyst in the canal of Nuck via the round ligament of the uterus. Though, many adult women with these types of inguinal hydrocoeles sometimes undergo fine-needle aspiration. CONCLUSION: This finding may highlight the need for a careful search for metastasis to the inguinal region in cases of ovarian cancer.

Idiopathic ventricular arrhythmias arising from the posterior septum of tricuspid and mitral annuli: comparison of electrocardiographic characteristics and outcomes of catheter ablation.

PURPOSE: Ventricular arrhythmia (VA) arising from the tricuspid annulus in the posterior septum (PS) (TAPS-VA) has similar electrocardiographic characteristics as VA arising from the mitral annulus in the PS (MAPS-VA) because the two locations are adjacent. We examined the electrocardiographic characteristics that differentiate MAPS-VA from TAPS-VA and the efficacy of catheter ablation. METHODS: We studied 13 patients whose VAs were successfully ablated in the TAPS (n = 7) and MAPS (n = 6). RESULTS: QRS morphologies of both groups were characterized by left and right bundle block morphologies in lead V1, superior axis deviation, and precordial transition at ≤ lead V3 in nine patients. Compared with TAPS-VA, MAPS-VA had (1) R < S wave in lead II, (2) precordial transition in lead V2, (3) s-wave in lead V6, and (4) slurred initial part of the QRS complex in the precordial leads, e.g., [4a] pseudo delta wave ≥ 34 ms, [4b] intrinsicoid deflection time ≥ 85 ms, and [4c] maximum deflection index ≥ 0.55. Patients who met ≥ 2 of (1)-(3) and any of [4a]-[4c] could be classified as having MAPS-VA, with a sensitivity and specificity of 100%. VA recurred in one patient in the TAPS group during the median follow-up of 7 (interquartile range 6 to 15.5) months. CONCLUSIONS: VA arising from the PS has superior axis deviation, and left and right bundle block morphologies with relatively early precordial transition. MAPS-VA can be differentiated from TAPS-VA based on electrocardiographic characteristics. This study provides useful information for treatment involving catheter ablation for VA arising from the PS.

Configuration of Materially Retained Carbon in Our Society: A WIO-MFA-Based Approach for Japan.

To achieve the goals of Paris Agreement, global society is directing much effort in substantially reducing greenhouse gas (GHG) emissions. In addition to energy-related efforts, prevention of carbon release into the atmosphere with carbon capture and storage (CCS) and/or utilization of biomass resources is considered indispensable to achieving the global objective. In this study, considering carbon-containing goods as carbon reservoirs in our society similar to forests and reservoirs enabling CCS, the flow of materially utilized carbon was quantified by input-output-based material flow analysis (IO-MFA). As a result, in 2011, 6.3 Mt-C of petroleum-derived carbon and 7.9 Mt-C of wood-derived carbon were introduced to the Japanese society as end-use products (e.g., automobiles and constructions) in various forms (e.g., plastics and synthetic rubbers). The total amount (14.2 Mt-C) corresponded to 4.1% (52.1 Mt-CO2) of annual CO2 emission in Japan in 2011. Subsequently, by referring to the technology that can treat carbon in the target forms in end-of-life products, the recoverability of carbon as a material has been discussed with respect to each form and end-use of carbon. By numerically showing the necessity and potential of implementing appropriate technologies, this study provides scientific direction for policymakers to establish a quality carbon cycle in our society.

Methicillin-Resistant Staphylococcus aureus Nasal Swab and Suction Drain Tip Cultures in 4573 Spinal Surgeries: Efficacy in Management of Surgical Site Infections.

STUDY DESIGN: A retrospective single-center study. OBJECTIVE: To assess the diagnostic value of methicillin-resistant Staphylococcus aureus (MRSA) nasal swab and suction drain tip cultures. SUMMARY OF BACKGROUND DATA: The prognostic value of MRSA nasal swab and suction drain tip cultures has not been firmly established in spinal surgery. METHODS: This study retrospectively included 4573 consecutive patients who underwent spinal surgery between January 2008 and December 2014. Patients diagnosed with infectious disease were excluded. Prophylactic antibiotics were administered intraoperatively and postoperatively for 48 hours. MRSA nasal swab cultures were taken from all patients before surgery. Drains were removed when the volume of postoperative fluid drainage was less than 50 mL in the preceding 24 hours and cultures were made. Surgical site infection (SSI) was defined according to Centers for Disease Control and Prevention criteria. RESULTS: SSI was identified in 94 cases (2.1%) and bacteria were isolated in 87 cases (92.6%). Positive MRSA nasal swab cultures were identified in 49 cases (1.1%). There was no significant difference in the SSI positivity rate between the MRSA nasal swab culture (+) and (-) groups. Positive drain tip cultures were found in 382 cases (8.4%), 28 of which developed SSI. There was a significant difference in the SSI positivity rate between the drain tip culture (+) and (-) groups. The sensitivity of drain tip culture was 29.8% and the specificity was 92.1%. In 16 of the 28 patients in the SSI (+) group with positive drain cultures, the same bacteria were isolated from the surgical site, giving a bacteria matching rate of 57.1%. CONCLUSION: MRSA nasal swab and drain tip cultures were not useful for predicting SSI. However, drain tip culture had a high positivity rate in the SSI group and the coincidence rate for the causative pathogen was relatively high. LEVEL OF EVIDENCE: 4.

Guidelines for the treatment of childhood-onset Graves' disease in Japan, 2016.

Purpose behind developing these guidelines: Over one decade ago, the "Guidelines for the Treatment of Graves' Disease with Antithyroid Drug, 2006" (Japan Thyroid Association (JTA)) were published as the standard drug therapy protocol for Graves' disease. The "Guidelines for the Treatment of Childhood-Onset Graves' Disease with Antithyroid Drug in Japan, 2008" were published to provide guidance on the treatment of pediatric patients. Based on new evidence, a revised version of the "Guidelines for the Treatment of Graves' Disease with Antithyroid Drug, 2006" (JTA) was published in 2011, combined with the "Handbook of Radioiodine Therapy for Graves' Disease 2007" (JTA). Subsequently, newer findings on pediatric Graves' disease have been reported. Propylthiouracil (PTU)-induced serious hepatopathy is an important problem in pediatric patients. The American Thyroid Association's guidelines suggest that, in principle, physicians must not administer PTU to children. On the other hand, the "Guidelines for the Treatment of Graves' Disease with Antithyroid Drug, 2011" (JTA) state that radioiodine therapy is no longer considered a "fundamental contraindication" in children. Therefore, the "Guidelines for the Treatment of Childhood-Onset Graves' Disease with Antithyroid Drug in Japan, 2008" required revision.

Uric acid ameliorates indomethacin-induced enteropathy in mice through its antioxidant activity.

BACKGROUND AND AIM: Uric acid is excreted from blood into the intestinal lumen, yet the roles of uric acid in intestinal diseases remain to be elucidated. The study aimed to determine whether uric acid could reduce end points associated with nonsteroidal anti-inflammatory drug (NSAID)-induced enteropathy. METHODS: A mouse model of NSAID-induced enteropathy was generated by administering indomethacin intraperitoneally to 8-week-old male C57BL/6 mice, and then vehicle or uric acid was administered orally. A group of mice treated with indomethacin was also concurrently administered inosinic acid, a uric acid precursor, and potassium oxonate, an inhibitor of uric acid metabolism, intraperitoneally. For in vitro analysis, Caco-2 cells treated with indomethacin were incubated in the presence or absence of uric acid. RESULTS: Oral administration of uric acid ameliorated NSAID-induced enteropathy in mice even though serum uric acid levels did not increase. Intraperitoneal administration of inosinic acid and potassium oxonate significantly elevated serum uric acid levels and ameliorated NSAID-induced enteropathy in mice. Both oral uric acid treatment and intraperitoneal treatment with inosinic acid and potassium oxonate significantly decreased lipid peroxidation in the ileum of mice with NSAID-induced enteropathy. Treatment with uric acid protected Caco-2 cells from indomethacin-induced oxidative stress, lipid peroxidation, and cytotoxicity. CONCLUSIONS: Uric acid within the intestinal lumen and in serum had a protective effect against NSAID-induced enteropathy in mice, through its antioxidant activity. Uric acid could be a promising therapeutic target for NSAID-induced enteropathy.

Current Status of Uterine Leiomyosarcoma in the Tohoku Region: Results of the Tohoku Translational Center Development Network Survey.

BACKGROUND: To prepare for a future clinical trial for improving the long-term prognosis of patients with uterine leiomyosarcoma (ULMS), we conducted a multi-institutional survey in the Tohoku region of Japan. METHODS: We conducted a retrospective cohort study between 2011 and 2014 in member institutions of the Tohoku Translational Research Center Development Network. RESULTS: A total of 53 patients with ULMS were registered in 31 institutions for the present survey. The median patient age was 56 years, 67.9% of the patients were postmenopausal, 88.7% had a performance status of 0 or 1, and only 6 patients (11.3%) showed preoperative evidence of malignancy. Although retroperitoneal lymphadenectomy was performed in only 26.4% of patients, 64.2% patients were identified as having FIGO stage 1 disease; 73.6% were eligible to undergo complete surgery. Among 36 patients who were treated with postoperative chemotherapy, 28 (77.8%) received docetaxel and gemcitabine combination therapy. The most frequent recurrence site was the lungs, and the median progression-free survival of all enrolled patients was 11.7 months. However, the median progression-free survival and the median overall survival in patients with stages III and IV disease were 3.4 and 11.4 months, respectively. CONCLUSION: Although ULMS was associated with a high rate of complete or optimal surgery, the long-term prognosis was poor. Effective postoperative therapy should be developed to improve the long-term prognosis of patients with ULMS.

A Comparative Study of Anterior Decompression With Fusion and Posterior Decompression With Laminoplasty for the Treatment of Cervical Spondylotic Myelopathy Patients With Large Anterior Compression of the Spinal Cord.

STUDY DESIGN: This is a retrospective observational single-center study. OBJECTIVES: To compare anterior decompression and fusion (ADF) and laminoplasty (LAMP) for the treatment of cervical spondylotic myelopathy (CSM) patients with large anterior compression in terms of clinical and radiologic outcomes. SUMMARY OF BACKGROUND DATA: We have reported that insufficient posterior decompression could be often seen after laminoplasty for CSM patients with preoperative anterior clearance of the spinal cord, defined as an interval <4 mm between the preoperative the modified K-line and anterior structure of the spinal canal at most compressive segment on sagittal T1-weighted magnetic resonance imaging. Here we conduct a study comparing ADF and LAMP for the treatment of CSM patients with such a risk factor. MATERIALS AND METHODS: Of the 221 consecutive CSM patients treated with either ADF or LAMP between 2008 and 2012 at our hospital, 79 patients in whom the interval was <4 mm with age ranged from 50 to 79 years were enrolled. Patients with myelopathy caused by single-level disk herniation, tumor or ossification of posterior longitudinal ligament, or patients with a history of cervical spine injury were excluded. The Japanese Orthopedic Association (JOA) scoring system for cervical myelopathy, recovery rate of the JOA score at the time of 2 years after surgery were investigated as clinical outcomes to compare these 2 groups. RESULTS: Demographics were almost similar between ADF and LAMP groups. The mean preoperative and postoperative JOA scores were 10.9 and 13.8 points for ADF group and 10.1 and 12.4 points for LAMP group, indicating that the recovery rate of JOA score was significantly greater in ADF group (49.6%) than that in LAMP group (38.2%; P=0.047). In LAMP group, spinal cord deformity was a significant predictive factor for unsatisfactory clinical outcome. CONCLUSION: ADF provided better surgical treatment for the patients with absence of preoperative anterior clearance of the spinal cord.

Psychological stress exacerbates NSAID-induced small bowel injury by inducing changes in intestinal microbiota and permeability via glucocorticoid receptor signaling.

BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) are popular painkillers, but they have serious side effects, not only in the upper gastrointestinal tract but also in the small intestine. It is well known that psychological stress may exacerbate various gastrointestinal diseases. The aim of this study was to determine whether psychological stress exacerbates NSAID enteropathy and to determine the possible underlying mechanisms for this. METHODS: Experiment 1: mice were exposed to water avoidance stress (WAS) or sham stress for 1 h per day for 8 consecutive days, and then enteropathy was induced by indomethacin. Experiment 2: cecal contents from stress (-) or (+) mice were transplanted into mice that had received antibiotics and in which NSAID enteropathy had been induced without WAS. Experiment 3: mifepristone, a glucocorticoid receptor antagonist, was injected before WAS for 8 days. Small intestinal injury, mRNA expression of TNFα, intestinal permeability, and the microbial community were assessed. RESULTS: Psychological stress exacerbated NSAID enteropathy and increased intestinal permeability. Psychological stress induced changes in the ileal microbiota that were characterized by increases in the total number of bacteria and the proportion of Gram-negative bacteria. The increased susceptibility to NSAIDs and intestinal permeability due to WAS was transferable via cecal microbiota transplantation. The increased permeability and aggravation of NSAID enteropathy caused by WAS were blocked by the administration of mifepristone. CONCLUSIONS: This study demonstrated a relationship between NSAID enteropathy and psychological stress, and showed the utility of studying the intestinal microbiota in order to elucidate the pathophysiology of NSAID enteropathy. It also showed the impact of stress on the intestinal microbiota and the mucosal barrier in gastrointestinal diseases.

Human serum albumin hydropersulfide is a potent reactive oxygen species scavenger in oxidative stress conditions such as chronic kidney disease.

Recently, hydropersulfide (RSSH) was found to exist in mammalian tissues and fluids. Cysteine hydropersulfide can be found in free cysteine residues as well as in proteins, and it has potent antioxidative activity. Human serum albumin (HSA) is the most abundant protein in mammalian serum. HSA possesses a free thiol group in Cys-34 that could be a site for hydropersulfide formation. HSA hydropersulfide of high purity as a positive control was prepared by treatment of HSA with Na2S. The presence of HSA hydropersulfide was confirmed by spectroscopy and ESI-TOFMS analysis where molecular weights of HSA hydropersulfide by increments of approximately 32 Da (Sulfur atom) were detected. The fluorescent probe results showed that Alexa Fluor 680 conjugated maleimide (Red-Mal) was a suitable assay and bromotrimethylammoniumbimane bromide appeared to be a selective reagent for hydropersulfide. The effect of oxidative stress related disease on the existence of albumin hydropersulfides was examined in rat 5/6 nephrectomy model of chronic kidney disease (CKD). Interestingly, the level of hydropersulfides in rat 5/6 nephrectomy model serum was decreased by a uremic toxin that increases oxidative stress in rat 5/6 nephrectomy model. Furthermore, we demonstrated that the levels of HSA hydropersulfide in human subjects were reduced in CKD but restored by hemodialysis using Red-Mal assay. We conclude that HSA hydropersulfide could potentially play an important role in biological anti-oxidative defense, and it is a promising diagnostic and therapeutic marker of oxidative diseases.

Antibiotics Suppress Activation of Intestinal Mucosal Mast Cells and Reduce Dietary Lipid Absorption in Sprague-Dawley Rats.

BACKGROUND & AIMS: The gut microbiota affects intestinal permeability and mucosal mast cells (MMCs) responses. Activation of MMCs has been associated with absorption of dietary fat. We investigated whether the gut microbiota contributes to the fat-induced activation of MMCs in rats, and how antibiotics might affect this process. METHODS: Adult male Sprague-Dawley rats were given streptomycin and penicillin for 4 days (n = 6-8) to reduce the abundance of their gut flora, or normal drinking water (controls, n = 6-8). They underwent lymph fistula surgery and after an overnight recovery were given an intraduodenal bolus of intralipid. We collected intestinal tissues and lymph fluid and assessed activation of MMCs, intestinal permeability, and fat transport parameters. RESULTS: Compared with controls, intestinal lymph from rats given antibiotics had reduced levels of mucosal mast cell protease II (produced by MMCs) and decreased activity of diamine oxidase (produced by enterocytes) (P < .05). Rats given antibiotics had reduced intestinal permeability in response to dietary lipid compared with controls (P < .01). Unexpectedly, antibiotics also reduced lymphatic transport of triacylglycerol and phospholipid (P < .01), concomitant with decreased levels of mucosal apolipoproteins B, A-I, and A-IV (P < .01). No differences were found in intestinal motility or luminal pancreatic lipase activity between rats given antibiotics and controls. These effects were not seen with an acute dose of antibiotics or 4 weeks after the antibiotic regimen ended. CONCLUSIONS: The intestinal microbiota appears to activate MMCs after the ingestion of fat in rats; this contributes to fat-induced intestinal permeability. We found that the gut microbiome promotes absorption of lipid, probably by intestinal production of apolipoproteins and secretion of chylomicrons.

Platelet interaction with lymphatics aggravates intestinal inflammation by suppressing lymphangiogenesis.

Lymphatic failure is a histopathological feature of inflammatory bowel disease (IBD). Recent studies show that interaction between platelets and podoplanin on lymphatic endothelial cells (LECs) suppresses lymphangiogenesis. We aimed to investigate the role of platelets in the inflammatory process of colitis, which is likely to be through modulation of lymphangiogenesis. Lymphangiogenesis in colonic mucosal specimens from patients with IBD was investigated by studying mRNA expression of lymphangiogenic factors and histologically by examining lymphatic vessel (LV) densities. Involvement of lymphangiogenesis in intestinal inflammation was studied by administering VEGF-receptor 3 (VEGF-R3) inhibitors to the mouse model of colitis using dextran sulfate sodium and evaluating platelet migration to LVs. The inhibitory effect of platelets on lymphangiogenesis was investigated in vivo by administering antiplatelet antibody to the colitis mouse model and in vitro by coculturing platelets with lymphatic endothelial cells. Although mRNA expressions of lymphangiogenic factors such as VEGF-R3 and podoplanin were significantly increased in the inflamed mucosa of patients with IBD compared with those with quiescent mucosa, there was no difference in LV density between them. In the colitis model, VEGF-R3 inhibition resulted in aggravated colitis, decreased lymphatic density, and increased platelet migration to LVs. Administration of an antiplatelet antibody increased LV densities and significantly ameliorated colitis. Coculture with platelets inhibited proliferation of LECs in vitro. Our data suggest that despite elevated lymphangiogenic factors during colonic inflammation, platelet migration to LVs resulted in suppressed lymphangiogenesis, leading to aggravation of colitis by blocking the clearance of inflammatory cells. Modulating the interaction between platelets and LVs could be a new therapeutic means for treating IBD.