Multiple intra-articular injections with adipose-derived stem cells for knee osteoarthritis cause severe arthritis with anti-histone H2B antibody production.

Introduction: Osteoarthritis (OA) is the most common form of arthritis. OA results from the breakdown of cartilage, which leads to deterioration of the entire joint and the connective tissue that holds the joint together, and gradually and irreversibly worsens over time. Adipose-derived stem/stromal cells (ADSCs) have been used in the treatment of knee OA. However, the safety and efficacy of ADSC treatment of OA remain unclear. In this study, we investigated the pathophysiology of severe knee arthritis that occurred after ADSC treatment by screening for autoantibodies in synovial fluid from patients who received ADSC treatment. Methods: Adult Japanese patients with OA who received ADSC treatment at Saitama Cooperative Hospital between June 2018 and October 2021 were enrolled. Antibodies (Abs) were screened using immunoprecipitation (IPP) with [35S]-methionine-labeled HeLa cell extracts. The detected protein was identified by liquid chromatography coupled with time-of-flight mass spectrometry (MS) and ion trap MS, and the corresponding proteins were confirmed as autoantigens using immunoblotting. Ab titers were measured using an enzyme-linked immunosorbent assay. Results: A total of 113 patients received ADSC treatment, and 75% (85/113) received ADSC injection at least twice with a 6-month interval between. No obvious abnormalities were observed in any patient after their first treatment; by contrast, 53% (45/85) of patients who received their second or third ADSC injection showed severe knee arthritis. IPP detected a common anti-15 kDa Ab in synovial fluid of 62% (8/13) of the samples analyzed from patients who showed severe arthritis. This Ab was not detected in synovial fluid obtained from the same joints before treatment. The corresponding autoantigen was identified as histone H2B. All available synovial samples from patients who tested positive for anti-histone H2B Ab were newly positive after the treatment; that is, none had been positive for anti-histone H2B Ab before treatment. Conclusions: Multiple ADSC injections for OA induced severe arthritis in a high percentage of patients, particularly after the second injection. Synovial fluid from some patients with knee arthritis contained Ab to histone H2B that appeared only after ADSC treatment. These findings provide new insights into the pathogenesis of ADSC treatment-induced severe arthritis.

Optimisation of Radiation Therapy in Bladder Preservation Therapy for Patients With Clinical Stage T2N0M0 Bladder Cancer.

AIMS: A novel bladder preservation therapy, the OMC (Osaka Medical College) regimen, which combines radiation therapy with balloon-occluded arterial infusion of anticancer agents, is a treatment option for patients with muscle-invasive bladder cancer (MIBC). We retrospectively analysed the effects of changes in radiation dose and irradiation field on treatment efficacy and adverse events.The purpose of this study is to use the results of this study to help determine a course of radiation therapy for bladder preservation therapy of cT2N0M0 MIBC. MATERIALS AND METHODS: We examined 352 patients with clinical stage T2N0M0 (cT2N0M0) MIBC classified into the following groups based on the irradiation method: group A, the whole pelvis (50 Gy/25 fractions) + local bladder (10 Gy/5 fractions); group B, the small pelvis (50 Gy/25 fractions) + local bladder (10 Gy/5 fractions); group C, the whole pelvis (40 Gy/20 fractions) + local bladder (10 Gy/5 fractions). RESULTS: The complete response rate, 3-year overall survival and progression-free survival rates in group A were 92.9%, 94.9% and 82.1%, respectively; in group B were 87.2%, 86.7% and 76.7%, respectively; and in group C were 95.2%, 92.6% and 71.1%, respectively. No significant differences between the groups were noted. The incidence of ≥grade 3 urinary tract and gastrointestinal toxicities were not significantly different among the groups (group A: 7.8%, 1.7%; B, 11.1%, 0%; C, 7.1%, 1.8%, respectively). The 3-year progression-free rates of the common iliac lymph node (CILN) region in patients who received whole-pelvis and small-pelvis irradiation were 99.0 and 89.0% (P < 0.01), respectively, with the latter group having significantly high lymph node recurrence in the CILN region. CONCLUSIONS: Our findings showed that the optimal radiation therapy for patients with cT2N0M0 MIBC undergoing the OMC regimen is whole-pelvis irradiation including the CILN region, with a total dose of 50 Gy/25 fractions.

Statistical mechanical determination of nanocluster size distributions in the phase coexistence region of a first order phase transition from the isotherms of DMPC monolayers at the air-water interface.

A statistical mechanical deconvolution procedure for the experimentally measured surface pressure-area isotherms has been presented to obtain the surface pressure dependence of the liquid expanded (LE) and liquid condensed (LC) nanocluster size distributions in the LE-LC phase coexistence region of the first order phase transition of Dimyristoyl phosphatidylcholine (DMPC) monolayers at the air-water interface. This study presents the application of the deconvolution formulation introduced originally by Freire and Biltonen for the experimentally measured specific heat to calculate the submicroscopic lipid cluster distribution function in the phase coexistence region [E. Freire, R. L. Biltonen, Biopolymers, 1978, 17, 481-496] and extends their formulation to surface pressure isotherms. The present procedure involves the extraction of the pressure partition function calculated from the isotherm and utilizes the general relation between molecular density fluctuations and macroscopic lateral compressibility. In this procedure the high-density LC phase boundary has been determined uniquely. The average nanoscopic cluster sizes obtained in this study have been compared with the results from previous experimental studies. The cause of the finite difference between the values of the LC phase boundary area obtained from the present deconvolution procedure and the conventional extrapolation method on the same isotherm has been discussed from the viewpoint of slow hierarchical growth from nanoscopic clusters to macroscopic domains in the coexistence region.

Long-term results of salvage photodynamic therapy for patients with local failure after chemoradiotherapy for esophageal squamous cell carcinoma.

BACKGROUND AND STUDY AIMS: Local failure after chemoradiotherapy (CRT) remains a major problem for patients with esophageal squamous cell carcinoma (ESCC). The aim of this study was to clarify the long-term results of salvage photodynamic therapy (PDT) for local failure. PATIENTS AND METHODS: Patients were treated with CRT, consisting of more than 50 Gy irradiation and concurrent chemotherapy. The indications for salvage PDT were as follows: 1) absence of lymph-node or distant metastasis after CRT; 2) failure lesion limited to T2; 3) refusal by patient to undergo salvage esophagectomy; 4) written informed consent. PDT was performed using an excimer dye laser at 48 and 72 hours after administration of Photofrin. RESULTS: A total of 37 consecutive patients underwent salvage PDT. The baseline stage before CRT was as follows: T1/T2/T3/T4 in 3/4/24/6 and N0/1 in 13/24 patients, respectively. Prior to PDT, 20 patients had a uT1 lesion, and 17 had a uT2 lesion; 24 patients had histologically proven local failure. A complete response was achieved in 22 patients (59.5%) following PDT. Esophageal fistulae, stenosis, and phototoxicity occurred in 4 (10.8%), 20 (54.1%), and 2 (5.4%) patients, respectively. Over a median follow-up period of 55 months, the 5-year progression-free (PFS) and overall survival rates of 37 patients following PDT were 20.7% and 36.1%, respectively. The 5-year PFS and overall survival of 24 patients with proven local failure were 17.6% and 34.6%, respectively. CONCLUSION: Salvage PDT is a curative treatment option for patients with local failure after CRT for ESCC.

Submucosal tumor appearance is a useful endoscopic predictor of early primary-site recurrence after definitive chemoradiotherapy for esophageal squamous cell carcinoma.

Chemoradiotherapy (CRT) for esophageal cancer is disadvantageous because of a high locoregional failure rate. Detecting early small recurrent cancers at the primary site is necessary for potential salvage treatment. However, most endoscopists are inexperienced and therefore, a role for surveillance endoscopy after complete remission (CR) has not been established. We retrospectively evaluated serial surveillance endoscopic images from patients eventually proved to have primary-site recurrence in order to identify useful endoscopic features for early diagnosis. From January 2000 to December 2004, 303 patients with esophageal squamous cell carcinoma underwent definitive CRT, and 133 of them achieved CR. The surveillance endoscopic images stored at intervals of 1-3 months for the 16 patients with recurrence only at the primary tumor site and the 61 patients with no recurrence were collected for reexamination. Among 133 patients who achieved CR, 16 (12%) developed only local recurrence at the primary site. Thirteen of the 16 primary-site recurrent tumors (81%) appeared as submucosal tumors (SMT), with the remaining appearing as erosions or mild strictures. Of biopsy-proven recurrences, 81% were preceded by newly developed lesions such as SMT, erosions, or mild strictures detected by earlier surveillance endoscopies. For all 77 patients achieving CR with no metastasis, 86% of the evolving SMT with negative biopsies were eventually confirmed as cancer at later endoscopies. Thirteen of the 21 evolving lesions were subsequently confirmed as recurrent cancer. Early primary-site recurrence of esophageal cancer after a complete response to CRT is detectable with frequent endoscopic surveillance. SMT appearance is a useful endoscopic sign of early recurrence, as well as a predictor of subsequent diagnosis of recurrence.

Long-term results of salvage endoscopic mucosal resection in patients with local failure after definitive chemoradiotherapy for esophageal squamous cell carcinoma.

BACKGROUND AND STUDY AIMS: Local failure after definitive chemoradiotherapy (CRT) in patients with esophageal cancer remains one of the major problems in finding a cure. Endoscopic mucosal resection (EMR) is one treatment option when failure lesions are superficial. However, there are no relevant long-term survival data. The aim of this study was to clarify the long-term survival of salvage EMR. PATIENTS AND METHODS: Between January 1998 and March 2004, 289 patients with esophageal squamous cell carcinoma were treated with definitive CRT at the National Cancer Center Hospital East, Japan. Of these 289 patients, 21 patients with local failure without lymph-node or distant metastases were treated with salvage EMR. The technique of salvage EMR involved a strip biopsy method. We retrospectively analyzed the long-term survival data for the patients who underwent salvage EMR. RESULTS: At a median follow-up period of 54 months (range, 16-108 months), eight of 21 patients (38%) were alive with no recurrence and two patients had died from another disease but with no recurrence of esophageal cancer. Local recurrence after EMR was detected in four patients, with local and lymph-node recurrence in two patients, and lymph-node and/or distant metastases in five patients. The 5-year survival rate from the initiation of salvage EMR was 49.1%. There were no severe complications associated with EMR. CONCLUSION: EMR is one of the curative salvage treatment options for local failure after definitive CRT, if the failure lesion is superficial and there are no lymph-node or distant metastases.

Anti-Salmonella activity of (2E)-alkenals.

AIMS: This study investigated the effect of a series of naturally occurring aliphatic (2E)-alkenals against Salmonella choleraesuis subsp. choleraesuis ATCC 35640 and evaluated their antibacterial action. METHOD AND RESULTS: A homologous series of aliphatic (2E)-alkenals from C5-C13 were tested for their antibacterial activity against Salm. Choleraesuis. The antibacterial action of (2E)-alkenals against Salm. choleraesuis increases with increasing carbon chain length. (2E)-Dodecenal (C12) was the most effective against this food-borne bacterium with minimum bactericidal concentration (MBC) of 6.25 microg ml-1 (34 micromol l-1), followed by (2E)-undecenal (C11) with an MBC of 12.5 microg ml-1 (77 micromol l-1). The activity was found to correlate with the hydrophobic alkyl chain length from the hydrophilic aldehyde group. The time-kill curve study showed that (2E)-dodecenal was bactericidal against Salm. choleraesuis at any growth stage. CONCLUSIONS: The antibacterial activity of (2E)-alkenals against Salm. choleraesuis was found to correlate with the hydrophobic alkyl chain length. The conjugated double bond is not essential in eliciting the activity but is associated with increasing it. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of their easy availability and wide distribution in many edible plants, (2E)-alkenals can be used as anti-Salmonella agents.

Effect of sodium valproate on neuromagnetic responses to chromatic flicker: implication for photosensitivity.

The effect of oral administration of sodium valproate in normal subjects was evaluated using whole-scalp magnetoencephalography, with results compared to the effect of sodium valproate in photosensitive children. Neuromagnetic responses to 10 Hz equiluminant red-green and red-blue flicker were measured before and after 5 days of sodium valproate administration. For the red-green flicker, relative power spectra at the stimulus frequency (10 Hz) were attenuated with medication in most brain regions. However, for the red-blue flicker, the 10-Hz power in the occipital region was enhanced with medication, while it was reduced in other regions. These results qualitatively resembled those in photosensitive children. The present findings suggest that (1) combinational chromatic sensitivity can be a critical factor for cortical excitability, that (2) the effect of sodium valproate is qualitatively similar in normal and photosensitive subjects, and that (3) the effect of sodium valproate on cortical excitability is not simply to suppress the stimulus-synchronized occipital activity, but rather to inhibit the spread of cortical activity from the occipital region to other regions.

Efficient and versatile synthesis of novel 2alpha-substituted 1alpha,25-dihydroxyvitamin D(3) analogues and their docking to vitamin D receptors.

Novel 2alpha-substituted 1alpha,25-dihydroxyvitamin D(3) analogues with 2alpha-alkyl and 2alpha-hydroxyalkyl groups were systematically synthesized from D-xylose. Their conformation on binding to the ligand binding domain (LBD) of the vitamin D receptor was analyzed. It has been found that the 2alpha-hydroxypropyl group best fits the cavity of the LBD, and the binding activity is three times higher than that for the natural hormone.

Cloning of a new aquaporin (AQP10) abundantly expressed in duodenum and jejunum.

A new aquaporin (AQP10) was identified in human small intestine. This gene encoded a 264-amino-acid protein with high sequence identity with AQP3 (53%), 9 (52%), and 7 (43%). These AQPs constitute one subfamily of AQP family that is differentiated from the other subfamily of AQP (AQP0, 1, 2, 4, 5, 6, and 8) by sequence homology. Ribonuclease protection assay and Northern blotting demonstrated almost exclusive expression of AQP10 mRNA in the duodenum and jejunum. In situ hybridization localized it in absorptive jejunal epithelial cells. Xenopus oocytes expressing AQP10 exhibited an increased osmotic water permeability in a mercury-sensitive manner. Although AQP10 belongs to the AQP subfamily, which has been characterized by permeability to water and neutral solutes such as urea and glycerol, it was not permeable to urea nor glycerol. The specific expression of AQP10 suggests its contribution to the water transport in the upper portion of small intestine.

Immunolocalization of aquaporin-8 in rat digestive organs and testis.

Expression of aquaporin-8 mRNA has previously been shown in hepatocytes, pancreatic acinar cells, colon epithelial cells and seminiferous tubules of the testis in the rat by in situ hybridization technique. However, immunolocalization of this water channel has not yet been demonstrated. In the present study, the localization of immunoreactive aquaporin-8 and expression of the mRNA were examined in rat organs (cerebrum, cerebellum, eye, salivary gland, heart, lung, liver, pancreas, esophagus, stomach, jejunum, ileum, colon, testis, ovary, kidney, spleen and lymphnode) by immunohistochemistry using an antibody against aquaporin-8 and ribonuclease protection assay. Aquaporin-8 was distinctly immunolocalized on the apical membranes of pancreatic acinar cells and mucosal epithelium of the colon and jejunum. In the liver, the bile canalicular membrane of hepatocytes was immunostained. In the testis, immunoreactive aquaporin-8 was demonstrated on the luminal side of the seminiferous tubules. At high magnification, the peroxidase reaction products appeared on the ramified cytoplasmic membrane of Sertoli cells surrounding the residual bodies or spermatogenic cells. Specificity of the antibody was verified by Western blot analysis showing a minor approximately 28 kDa band (deduced deglycosylated form of aquaporin-8) and a major approximately 30 kDa band (glycosylated form) in these organs. The intensity of aquaporin-8 immunoreactivity was approximately comparable to that of aquaporin-8 mRNA expression in the liver, pancreas, colon, jejunum and testis. The aquaporin-8 mRNA expression in the hepatocytes was presumed to be closely associated with the structure of bile canaliculi since the message was detected in hepatocytes immediately after isolation from the liver but not in cells following cultivation for three days. The localization of immunoreactive aquaporin-8 indicated functions for this water channel in the secretion of bile and pancreatic juice, and the secretion or absorption of water in the colon and jejunum, and the maturation or liberation of spermatogenic cells in the testis.

Influence of postsurgical residual tumor volume on local control in radiotherapy for maxillary sinus cancer.

BACKGROUND: The aim was to study the influence of postsurgical gross residual tumor volume on local control of maxillary sinus cancer treated with radiotherapy combined with debulking surgery. METHODS: Forty-three patients who underwent combined surgery and radiotherapy (50-72 Gy, median 60 Gy) for squamous cell carcinoma of the maxillary sinus were reviewed. Gross residual tumor volume (GRTV) after surgery was measured on computed tomograms obtained during the radiotherapy planning. Patients were classified according to GRTV as follows: group AA, GRTV = 0 (microscopic residual, n = 2); group A, GRTV < 10 cm3 (n = 24); group B, 10-40 cm3 (n = 9); and group C, > or = 40 cm3 (n = 8). The relationship between local control and GRTV was analyzed using univariate and multivariate analysis. RESULTS: The 2-year local control rate for all patients was 62%. The differences in local control rates between groups AA, A and B were not significant (P > 0.05), but the rate was significantly lower in group C than in the other groups (69% at 2 years vs 31% at 1 year, P < 0.001). Multivariate analysis showed that GRTV (P = 0.002) and histological differentiation (poorly differentiated histology was favorable, P = 0.035) were independent prognostic factors and that intra-arterial chemotherapy and administered total dose were not. Local control in groups A and B significantly depended on the total dose of radiotherapy, with 2-year control rates of patients receiving 50 Gy (n = 6) and > or = 60 Gy (n = 27) of 17% vs 79%, respectively (P < 0.001). CONCLUSIONS: Our data suggest that adequate, not complete, debulking associated with a total radiotherapy dose of > or = 60 Gy can provide satisfactory local control for patients with squamous cell carcinoma of the maxillary sinus.

Immunolocalization of aquaporin-9 in rat hepatocytes and Leydig cells.

The aquaporin (AQP)-9 gene was recently isolated from human and rat liver cDNA libraries as a member of the water channel family for water and neutral solutes. Although the expression of AQP9 mRNA has been demonstrated in several organs including the liver and testis by Northern blot analysis, the cellular and subcellular localization of the AQP9 protein remains unclear. In the present light and electron microscopic immunohistochemical study, the localization of the AQP9 immunoreactivity was examined in fifteen kinds of rat organs using an antibody against rat AQP9 synthetic peptide. The antibody immunostained a major band of approximately 33 kDa in the liver by Western blot analysis. Immunoreactivity for AQP9 was found exclusively in the liver and testis among the organs examined. In the liver, positive staining appeared selectively along the space of Disse. Immunoelectron microscopy confirmed the localization of AQP9 on the surface of hepatocyte microvilli facing the space of Disse. In the testis, the plasma membrane of Leydig cells located between seminiferous tubules was conspicuously immunoreactive to the antibody. Intense mRNA expression was detected in the liver and testis but not in other organs by ribonuclease protection assay. These findings suggest a specific role for AQP9 in the transport of water and non-charged solutes in hepatocytes and Leydig cells.