The evaluation of the safety and efficacy of intravenously administered allogeneic multilineage-differentiating stress-enduring cells in a swine hepatectomy model.

INTRODUCTION: Multilineage-differentiating stress-enduring (Muse) cells are non-tumorigenic endogenous pluripotent-like cells residing in the bone marrow that exert a tissue reparative effect by replacing damaged/apoptotic cells through spontaneous differentiation into tissue-constituent cells. Post-hepatectomy liver failure (PHLF) is a potentially fatal complication. The main purpose of this study was to evaluate the safety and efficiency of allogeneic Muse cell administration via the portal vein in a swine model of PHLF. METHODS: Swine Muse cells, collected from swine bone marrow-mesenchymal stem cells (MSCs) as SSEA-3(+) cells, were examined for their characteristics. Then, 1 × 107 allogeneic-Muse cells and allogeneic-MSCs and vehicle were injected via the portal vein in a 70% hepatectomy swine model. RESULTS: Swine Muse cells exhibited characteristics comparable to previously reported human Muse cells. Compared to the MSC and vehicle groups, the Muse group showed specific homing of the administered cells into the liver, resulting in improvements in the control of hyperbilirubinemia (P = 0.04), prothrombin international normalized ratio (P = 0.05), and suppression of focal necrosis (P = 0.04). Integrated Muse cells differentiated spontaneously into hepatocyte marker-positive cells. CONCLUSIONS: Allogeneic Muse cell administration may provide a reparative effect and functional recovery in a 70% hepatectomy swine model and thus may contribute to the treatment of PHLF.

Novel Immunoliposome Technology for Enhancing the Activity of the Agonistic Antibody against the Tumor Necrosis Factor Receptor Superfamily.

We have developed a technology for efficiently enhancing the anticancer apoptosis-inducing activity of agonistic antibodies against the tumor necrosis factor receptor (TNFR) superfamily by the formation of immunoliposomes. To induce apoptosis in cancer cells, agonistic antibodies to the TNFR superfamily normally need cross-linking by internal immune effector cells via the Fc region after binding to receptors on the cell membrane. To develop apoptosis-inducing antibodies that do not require the support of cross-linking by immune cells, we prepared immunoliposomes conjugated with TRA-8, an agonistic antibody against death receptor 5 (DR5), with various densities of antibody on the liposome surface, and evaluated their activities. The TRA-8 immunoliposomes exhibited apoptosis-inducing activity against various DR5-positive human carcinoma cells at a significantly lower concentration without cross-linking than that of the original TRA-8 and its natural ligand (TRAIL). The activity of the immunoliposomes was correlated with the density of antibodies on the surface. As the antibody component, not only the full-length antibody but also the Fab' fragment could be used, and the TRA-8 Fab' immunoliposomes also showed exceedingly high activity compared with the parental antibody, namely, TRA-8. Moreover, cytotoxicity of the TRA-8 full-length or Fab' immunoliposome against normal cells, such as human primary hepatocytes, was lower than that for TRAIL. Enhanced activity was also observed for immunoliposomes conjugated with other apoptosis-inducing antibodies against other receptors of the TNFR superfamily, such as death receptor 4 (DR4) and Fas. Thus, immunoliposomes are promising as a new modality that could exhibit significant activity at a low dose, for cost-effective application of an antibody fragment and with stable efficacy independent of the intratumoral environment of patients as a TNF superfamily agonistic therapy.

Availability of 24-h urine collection method on dietary phosphorus intake estimation.

Accurate assessment of dietary phosphorus intake is necessary to prevent hyperphosphatemia. The aim of this study was to evaluate the 24-h urine collection method for estimation of phosphate intake in healthy males. Two experiments, a 1-day and a 5-day loading test, were performed. After an overnight fast, subjects consumed test meals, 24-h urine collection was performed, and blood samples were obtained. In the 5-day loading test, a phosphorus supplement was orally administered on day 3. The association between the phosphorus content of test meals and urinary excretion, anthropometric indices, and blood biomarkers was analyzed to develop a more precise formula for estimating phosphorus intake. In the 1-day loading test, the standard deviation of predictive phosphorus intake, based on multiple linear regression analysis, was less than that for the phosphorus absorption rate. In the 5-day loading test, urinary phosphorus excretion was similar on days 2, 4 and 5, but was significantly higher on day 3 after phosphorus supplementation. Our results indicate that estimation of dietary phosphorus intake with the 24-h urine collection method, using the amount of phosphorus and urea nitrogen excretion, may increase the precision of short-term monitoring.

A case of an intramesenteric GIST accurately diagnosed and completely resected by PET-CT and laparoscopy after gastrectomy for gastric cancer.

We performed the accurate diagnosis and complete surgical resection of a gastrointestinal stromal tumor at the mesentery of the small bowel. Computed tomography (CT) in a 62-year-old man at 2 years after gastrectomy for gastric cancer showed a mesenteric tumor, with no other tumors noted. Positron emission tomography-computed tomography (PET-CT) showed a maximum standardized uptake value (SUV max) of 2.9 at the tumor. The presence of a single and low SUV max tumor allowed us to perform laparoscopic surgery. Partial resection of the tumor with an adequate margin was performed. The pathological findings showed c-kit positivity and a low Ki-67 proliferation index (<5%). In the present case, PET-CT and laparoscopic assessments were useful for accurately evaluating the surgical resectability of the mesenteric tumor after distal gastrectomy for gastric cancer. The low SUV max and laparoscopic findings led to complete surgical resection of a mesenteric tumor.

Stromal expression of hemopexin is associated with lymph-node metastasis in pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma is one of the most aggressive types of cancer. Certain proteins in the tumor microenvironment have attracted considerable attention owing to their association with tumor invasion and metastasis. Here, we used proteomics to identify proteins associated with lymph-node metastasis, which is one of the prognostic factors. We selected lymph node metastasis-positive and -negative patients (n = 5 each) who underwent pancreatectomy between 2005 and 2015 and subjected to comprehensive proteomic profiling of tumor stroma. A total of 490 proteins were detected by mass spectrometry. Software analysis revealed that nine of these proteins were differentially expressed between the two patient groups. We focused on hemopexin and ferritin light chain based on immunohistochemistry results. We assessed the clinicopathological data of 163 patients and found that hemopexin expression was associated with UICC N2 (p = 0.0399), lymph node ratio (p = 0.0252), venous invasion (p = 0.0096), and lymphatic invasion (p = 0.0232). Notably, in vitro assays showed that hemopexin promotes invasion of the pancreatic cancer cells. Our findings suggest that hemopexin is a lymph node metastasis-associated protein that could potentially serve as a useful therapeutic target or biomarker of pancreatic ductal adenocarcinoma.

Cochlear implantation in a case of bilateral sensorineural hearing loss due to mumps.

It is well known that unilateral profound sensorineural hearing loss is caused by mumps; however, bilateral deafness is rare. Herein we report a case of bilateral profound hearing loss caused by mumps infection in a four-year-old boy. Labyrinthitis due to the mumps virus was suspected. His verbal understanding was poor, and he completely stopped talking. He was soon fitted with a hearing aid, but it proved insufficient. Thereupon, cochlear implantation was performed on his left ear. Six months after the operation, his speech perception and speech production were improved. In cases of bilateral profound hearing loss due to mumps infection conservative therapy is ineffective; therefore, cochlear implantation is recommended. Vaccine coverage for mumps virus is also strongly recommended in Japan.

Regeneration of the trachea using a bioengineered scaffold with adipose-derived stem cells.

OBJECTIVES: Our group has developed and clinically applied an artificial graft made from a collagen sponge scaffold for the regeneration of tracheal tissue. However, the artificial graft requires about 2 months for epithelial regeneration. The purpose of the present study was to accelerate the regeneration process of the trachea through the effective use of a bioengineered scaffold. Adipose-derived stem cells (ASCs) with multilineage differentiation capability were used. In our study, we implanted a bioengineered scaffold that included autologous ASCs into tracheal defects in rats. METHODS: Collagen gel, including ASCs labeled with monomeric yellow fluorescent protein, was layered onto the surface of the collagen sponge to form a bioengineered scaffold. This scaffold was implanted into the tracheal defects in rats. A control scaffold without ASCs was also implanted. RESULTS: On day 14 after implantation, a pseudostratified columnar epithelium with well-differentiated ciliated and goblet cells and neovascularization was observed in rats that received the implant with the bioengineered scaffold that included ASCs. CONCLUSIONS: These results suggested that implanted ASCs accelerated neovascularization and epithelialization on the regenerated trachea. Thus, our newly developed bioengineered scaffold contributes to tracheal regeneration.

A novel liver metastasis-correlated protein of pancreatic neuroendocrine neoplasm (PanNEN) discovered by proteomic analysis.

The aim of this study was to identify novel liver metastasis-correlated proteins of PanNEN by proteomics to compare pancreatic tumor (PT) with paired metastatic liver tumor (LT). Of 118 surgical cases with PanNEN, 7 cases with formalin-fixed paraffin-embedded (FFPE) tissues of both PT and paired LT were evaluated by proteomics. Tumor cells were selectively collected from FFPE tissues by laser capture microdissection. A total of 3,722 proteins were detected from extracted peptides by mass spectrometry-based shotgun analysis. Selection of the candidate proteins expressed differently between PT and LT were performed by semi-quantitative comparison in silico and confirmation with immunohistochemistry. We focused on ANXA6, CNPY2, RAB11B and TUBB3, all of which had higher expressions in LT. In all surgical cases with FFPE samples, liver recurrence-free survival (RFS) was evaluated in correlation to the expression of the candidate proteins in PT by immunohistochemistry. Liver RFS was significantly poorer in CNPY2 positive patients than in negative patients (10-year liver RFS; 39.8% vs. 92.3%, p = 0.012). Also, liver RFS tended to be poorer in ANXA6 positive patients than in those who were negative (10-year liver RFS; 51.4% vs. 95.0%, p = 0.099). In the multivariate analysis, the independent predictors of liver RFS were CNPY2 positivity (HR: 6.19, 95 % CI: 1.47-42.79, p = 0.011) and tumor size ≥ 42 mm (HR: 4.63, 95 % CI: 1.03-23.23, p = 0.045). In conclusion, CNPY2 is a novel liver metastasis-correlated protein of PanNEN.

Age-dependent degeneration of the stria vascularis in human cochleae.

OBJECTIVE: Aging is a common cause of acquired hearing impairments. This study investigated age-related morphologic changes in human cochleae, with a particular focus on degeneration of the stria vascularis (SV) and the spiral ganglion (SG). STUDY DESIGN: Retrospective case review. METHODS: The study group comprised 91 temporal bones from individuals aged 10 to 85 years who had no history or audiometric findings suggestive of specific causes of cochlear degeneration. We quantified the SV and SG atrophy at each cochlear turn using morphometric measurements. Correlations of the SV and SG atrophy with age, audiometric patterns of hearing loss, and auditory thresholds were statistically investigated. RESULT: The SV and the SG both showed a tendency for progressive atrophy to develop with age. However, statistically significant correlations were observed between aging and SV atrophy only in the apical and basal cochlear turns. These findings were consistent with those reported previously in gerbils. No significant correlations were detected between SV or SG atrophy and audiometric findings. CONCLUSION: SV atrophy appears to be the most prominent anatomic characteristic of aged human cochleae.

Presence of cytomegalovirus in the perilymphatic fluid of patients with profound sensorineural hearing loss caused by congenital cytomegalovirus infection.

CONCLUSION: Not all patients diagnosed with congenital infection using umbilical cord assay were found to be positive for CMV-DNA by perilymphatic fluid assay. In addition, a CMV-DNA-positive result was observed in one patient who had not been diagnosed with congenital infection. Sampling of perilymphatic fluid from a large population of patients with congenital SNHL caused by congenital CMV infection or of unknown etiology is required to determine the prevalence of CMV-related profound HL. OBJECTIVES: Sensorineural hearing loss (SNHL) is one of the most frequent manifestations in patients with congenital cytomegalovirus (CMV) infection. Using dried umbilical cord, a PCR-based assay was recently developed for the retrospective detection of congenital CMV infection. This study analyzed the presence of CMV in the perilymphatic fluid and evaluated differences in the effect of cochlear implantation between CMV-positive and -negative groups. METHOD: Perilymphatic fluid was collected from each patient at the time of cochlear implantation and analyzed for the presence of CMV using a PCR method. RESULTS: The perilymphatic fluid in two of the five patients suffering from congenital CMV infection and in one of the 17 patients without congenital CMV infection was found to be positive for CMV.

[Temporal bone study of residual mastoid inflammation in ears with normally appearing tympanic membrane].

We examined 395 temporal bones with an intact tympanic membrane to explore the relationship between residual inflammation in the middle ear cavity and development of mastoid pneumatization. Histopathological changes were studied in the middle ear cavity. Mastoid pneumatization was classified as good or poor based on the extent of mastoid tip development to the lateral semicircular canal. Specimens were 344 temporal bones with well-pneumatized mastoid and 51 with poorly-pneumatized mastoid. Otitis media was noted in 119 (34.6%) bones in the good group and 9 (17.6%) in the poor group. In well-pneumatized mastoid, chronic inflammatory changes were frequently observed at the lower portion of mastoid cells, the round window niche, and the tympanic sinus. In contrast, no such incidence of inflammatory change was noted in poorly-pneumatized mastoid. Our findings indicate that an intact tympanic membrane does not always mean freedom from mastoid inflammation, especially when the mastoid is well-pneumatized. This makes it important to check for possible remaining otitis media in patients with a well-pneumatized mastoid, even if the tympanic membrane appears normal.

Outcome of cochlear implantation in children with congenital cytomegalovirus infection or GJB2 mutation.

CONCLUSIONS: Outcomes following cochlear implantation in children with congenital cytomegalovirus (CMV) infection were almost equivalent to those of children with GJB2 mutation-related sensorineural hearing loss (SNHL). Although our patients with developmental disorder showed poor auditory performance and speech and language skills after cochlear implantation, SNHL with developmental disorder should not be a contraindication for the procedure. OBJECTIVE: Congenital CMV infection accounts for approximately 20% of all cases of neonatal hearing loss, while the GJB2 mutation accounts for 30-50% of all cases of profound nonsyndromic hearing loss. Here, outcomes for auditory behavior and speech and language skills were compared in children with congenital CMV infection or GJB2 mutation who received cochlear implantation for profound SNHL. METHODS: Five children with asymptomatic congenital CMV infection and seven children with GJB2 mutation-related SNHL, with and without developmental disorder, underwent cochlear implantation. Hearing level and speech and language development were evaluated post-implantation using IT-MAIS, MUSS, and S-S method. RESULTS: The IT-MAIS and MUSS scores of the congenital CMV infection group and the GJB2 mutation group continued to increase for 4 years after implantation. The S-S method score in both groups gradually increased, although the scores for children with mental retardation were low.

PEGylation of osteoprotegerin/osteoclastogenesis inhibitory factor (OPG/OCIF) results in decreased uptake into rats and human liver.

OBJECTIVES: Our aim was to investigate the effect of PEGylation on the uptake of osteoprotegerin/osteoclastogenesis inhibitory factor (OPG/OCIF) into rat liver, kidney and spleen, and human liver. METHODS: Copolymer of polyethyleneglycol allylmethylether and maleamic acid sodium salt with OCIF (poly(PEG)-OCIF) (0.5 mg/kg) was administered to rats and the concentrations of poly(PEG)-OCIF in the liver, kidney and spleen at 15 min after administration were measured by ELISA. For human liver uptake, the liver perfusion of OCIF and (3)H-labelled poly(PEG)-OCIF was conducted using fresh human liver block. KEY FINDINGS: The tissue uptake of poly(PEG)-OCIF in rats was significantly lower compared with that of OCIF. In fresh human liver perfusion, (3)H-poly(PEG)-OCIF was rarely taken up into the liver. On the other hand, more than 50% of the perfused OCIF was taken up. CONCLUSIONS: PEGylation of OCIF using poly(PEG) dramatically suppressed the uptake of OCIF into human liver as well as into rat liver and could be a promising approach for improving the pharmacokinetic and pharmacological effects of OCIF in the clinical setting.

A case of severe acute pancreatitis, in overlap syndrome of systemic sclerosis and systemic lupus erythematosus, successfully treated with plasmapheresis.

Here, we describe a case of severe pancreatitis in overlap syndrome of systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) in an adult female patient. Treatment with plasmapheresis and high-dose prednisone successfully saved her life and led to remission of the pancreatitis. This is the first case report of severe acute pancreatitis in the setting of overlap syndrome of SLE and SSc. The advantages and disadvantages of the use of corticosteroids are discussed.