Septin7 regulates inner ear formation at an early developmental stage.

Septins are guanosine triphosphate-binding proteins that are evolutionally conserved in all eukaryotes other than plants. They function as multimeric complexes that interact with membrane lipids, actomyosin, and microtubules. Based on these interactions, septins play essential roles in the morphogenesis and physiological functions of many mammalian cell types including the regulation of microtubule stability, vesicle trafficking, cortical rigidity, planar cell polarity, and apoptosis. The inner ear, which perceives auditory and equilibrium sensation with highly differentiated hair cells, has a complicated gross morphology. Furthermore, its development including morphogenesis is dependent on various molecular mechanisms, such as apoptosis, convergent extension, and cell fate determination. To determine the roles of septins in the development of the inner ear, we specifically deleted Septin7 (Sept7), the non-redundant subunit in the canonical septin complex, in the inner ear at different times during development. Foxg1Cre-mediated deletion of Sept7, which achieved the complete knockout of Sept7 within the inner ear at E9.5, caused cystic malformation of inner ears and a reduced numbers of sensory epithelial cells despite the existence of mature hair cells. Excessive apoptosis was observed at E10.5,E11.5 and E12.5 in all inner ear epithelial cells and at E10.5 and E11.5 in prosensory epithelial cells of the inner ears of Foxg1Cre;Septin7floxed/floxed mice. In contrast with apoptosis, cell proliferation in the inner ear did not significantly change between control and mutant mice. Deletion of Sept7 within the cochlea at a later stage (around E15.5) with Emx2Cre did not result in any apparent morphological anomalies observed in Foxg1Cre;Septin7floxed/floxed mice. These results suggest that SEPT7 regulates gross morphogenesis of the inner ear and maintains the size of the inner ear sensory epithelial area and exerts its effects at an early developmental stage of the inner ear.

Anti-adhesive effects of a newly developed two-layered gelatin sheet in dogs.

AIM: Adhesion after pelvic surgery causes infertility, ectopic pregnancy, and ileus or abdominal pain. The materials currently available for clinical use are insufficient. The purpose of this study was to develop an anti-adhesive material that overcomes the limitations of conventional anti-adhesive agents. METHODS: The adhesion prevention effects of three methods - a two-layered sheet composed of gelatin film and gelatin sponge, Seprafilm and INTERCEED - were evaluated in 37 dogs. Anti-adhesive effects were investigated macroscopically and microscopically in a cauterized uterus adhesion model. Cell growth on the materials in vitro using human peritoneal mesothelial cells, fibroblasts and uterine smooth muscle cells were also evaluated. RESULTS: The two-layered gelatin sheet had significantly superior anti-adhesive effects compared to the conventional materials (Seprafilm and INTERCEED). A single-cell layer of mature mesothelium formed three weeks after surgery in the gelatin group. Peritoneum regeneration in the Seprafilm and INTERCEED groups was delayed and incomplete in the early phase. Little inflammation around the materials occurred and cell growth was significantly proliferated with the gelatin sheet. CONCLUSION: The anti-adhesive effects of a two-layered gelatin sheet were superior to conventional agents in a cauterized canine uterus model, demonstrating early regeneration of the peritoneum, little inflammation and material endurance. The newly developed two-layered gelatin sheet is a useful option as an anti-adhesive agent for deeply injured and hemorrhagic sites.

Hedgehog signaling regulates prosensory cell properties during the basal-to-apical wave of hair cell differentiation in the mammalian cochlea.

Mechanosensory hair cells and supporting cells develop from common precursors located in the prosensory domain of the developing cochlear epithelium. Prosensory cell differentiation into hair cells or supporting cells proceeds from the basal to the apical region of the cochleae, but the mechanism and significance of this basal-to-apical wave of differentiation remain to be elucidated. Here, we investigated the role of Hedgehog (Hh) signaling in cochlear development by examining the effects of up- and downregulation of Hh signaling in vivo. The Hh effector smoothened (Smo) was genetically activated or inactivated specifically in the developing cochlear epithelium after prosensory domain formation. Cochleae expressing a constitutively active allele of Smo showed only one row of inner hair cells with no outer hair cells (OHCs); abnormal undifferentiated prosensory-like cells were present in the lateral compartment instead of OHCs and their adjacent supporting cells. This suggests that Hh signaling inhibits prosensory cell differentiation into hair cells or supporting cells and maintains their properties as prosensory cells. Conversely, in cochlea with the Smo conditional knockout (Smo CKO), hair cell differentiation was preferentially accelerated in the apical region. Smo CKO mice survived after birth, and exhibited hair cell disarrangement in the apical region, a decrease in hair cell number, and hearing impairment. These results indicate that Hh signaling delays hair cell and supporting cell differentiation in the apical region, which forms the basal-to-apical wave of development, and is required for the proper differentiation, arrangement and survival of hair cells and for hearing ability.

Autoamputation of a pedunculated, subserosal uterine leiomyoma presenting as a giant peritoneal loose body.

Peritoneal loose bodies (PLBs) are defined as fibrotic or calcified-free bodies within the peritoneal cavity; they commonly autoamputate from appendices epiploicae that have undergone torsion. Pedunculated, subserosal uterine leiomyomas (PSULs) are subserosal uterine leiomyomas connected to the uterus via a pedicle. In the present report, we describe the case of a PLB that originated from the autoamputation of a PSUL, confirmed based on histological evidence consistent with a uterine leiomyoma and the laparoscopic findings of a broken pedicle. This case clearly demonstrates the potential for a uterine leiomyoma to be the source of a PLB. Our findings contribute to the understanding of the etiological relationship between PLBs and uterine leiomyomas.

Development of gelatin flakes, a new type of anti-adhesive material: a preliminary study of in vivo rat adhesion models.

To overcome the problems associated with sheet- or film-type anti-adhesive materials, we developed a new type of anti-adhesive material, gelatin flakes. We made two types of gelatin flakes with or without thermal cross-linking, and preliminarily examined their basic properties and the anti-adhesive efficacy using a rodent adhesion model. Both types of the gelatin flakes rapidly turned into gel and tightly attached the injured surfaces, absorbing the moisture and blood, when applied onto the abraded sites of rats. In addition, these flakes could be sprayed into the desired area by compressed air through a device with a long, thin tube, which could be used in laparoscopic surgery. The anti-adhesive effects of both types of gelatin flakes were similar, and both types were significantly superior compared to the non-treated group. Although further investigations are necessary, the gelatin flakes have unique and useful properties and satisfactory anti-adhesive effects, which indicate that they may be applicable in laparoscopic surgery.

Characteristics of Epithelioid Trophoblastic Tumor: Endoscopic and Magnetic Resonance Imaging Findings.

Introduction: Epithelioid endothelial tumor (ETT) is an extremely rare tumor that typically occurs in women of reproductive age. The diagnosis tends to be delayed because it often necessitates a total hysterectomy. Therefore, it is important to understand ETT macroscopic and imaging findings. Here, we report a case of ETT with detailed macroscopic and imaging findings. Case Presentation: A 39-year-old woman with positive pregnancy test results was admitted to a nearby hospital. No gestational sac was found in the uterus, and magnetic resonance imaging (MRI) revealed a cystic mass of approximately 7 cm that extended continuously from the anterior wall of the lower uterine segment into the pelvic cavity. She underwent laparoscopic and hysteroscopic surgeries for a ruptured cervical pregnancy. Pathology of the specimens obtained from this surgery did not allow for the diagnosis of ETT. Two months after the surgery, as the serum human chorionic gonadotropin ß subunit (ß-HCG) level did not decrease, she was diagnosed with low-grade gestational trophoblastic neoplasia, leading to the administration of chemotherapy. After three regimens of chemotherapy over 9 months, her ß-HCG level decreased but did not reach normal levels. Ultimately, a total hysterectomy was performed. The pathological diagnosis was mixed ETT and choriocarcinoma. A literature review revealed several cases similar to ours. Conclusion: ETT in the lower uterus often perforates the myometrium and forms cystic lesions in the retroperitoneal space or subserosa. The MRI and laparoscopic/hysteroscopic findings in this case may have contributed to the early diagnosis of ETT.

Short term-efficacy and tolerability of Rheocarna, a novel direct hemoperfusion adsorptive column, for chronic limb-threatening ischemia in dialysis patients: A single-center case series.

INTRODUCTION: With population aging and lifestyle changes, the number of patients with chronic limb-threatening ischemia (CLTI) is increasing, and refractory or recurrent lesions are more common, especially in chronic dialysis patients. In March 2021, a new type of adsorptive cellulose bead column immobilized with dextran sulfate and L-tryptophan for direct hemoperfusion (DHP) was approved by Japan's medical insurance system as a treatment for CLTI. METHODS: We retrospectively analyzed 17 cases of CLTI in dialysis patients treated with DHP using the novel column (Rheocarna) (DHP-R) at our hospital from May 2021 to October 2022. The short-term of efficacy of DHP-R was judged qualitatively by the foot care team every 2 weeks based on the assessment of skin color, warmth, ulcer epithelialization or shrinkage of the ulcer area, and foot pain. The final judgment of efficacy was made after the final DHP-R session. RESULTS: The median age of patients was 66 years, the median dialysis duration was 10 years, 15 cases (88%) were male, and 15 cases (88%) had diabetes. The median total number of sessions was eight. In comparing the groups in which DHP-R was effective and ineffective, there was no significant difference in any factors including patient background data (i.e., age, diabetes, low-density lipoprotein cholesterol, hemoglobin, dialysis duration, etc.), type of anticoagulants, and presence of episodes of blood pressure drop or circuit clotting during session. Three cases with symptomatic hypotension during the session and two cases with circuit clotting that did not improve with increased heparin dose all resolved immediately after changing the anticoagulant from heparin to nafamostat mesylate (NM). CONCLUSION: Identification of patients' characteristics in which DHP-R is favorable and some reliable index that allow a rapid decision to continue DHP-R are needed. In addition, validating whether the use of NM as anticoagulant affects the efficacy of DHP-R for CTLI treatment remains a challenge to resolve.

Eosinophilic airway inflammation and eosinophilic chronic rhinosinusitis during nivolumab and ipilimumab.

Immune-related adverse events (irAEs) are induced by immune checkpoint inhibitors (ICIs) which are administered for many cancers. There are many irAEs such as endocrine abnormalities, interstitial lung disease, and colitis. However, irAEs associated with type 2 (T2) inflammation are less known. We herein report a 71-year-old woman who developed eosinophilic airway inflammation and eosinophilic chronic rhinosinusitis (ECRS) simultaneously during combination therapy with nivolumab and ipilimumab for renal cell carcinoma. After two cycles of therapy, she developed cough and nasal congestion with high level of fractioned exhaled nitric oxide and blood eosinophil count, and nasal polyps with eosinophil infiltration in bilateral nasal cavities. She was diagnosed with eosinophilic airway inflammation and ECRS, and treated with corticosteroid inhalation, steroid nasal spray, and nasal irrigation, resulting in symptom reduction. Although they are relatively rare irAEs of ICIs, clinicians should consider these diseases associated with T2 inflammation and treat appropriately.

New polyglycolic acid fabric for the prevention of postoperative pancreatic fistulas.

BACKGROUND: The incidence of postoperative pancreatic fistula (POPF) after distal pancreatectomy is approximately 30%. The most serious complications of pancreatic resection, such as mortality and prolonged hospitalization, are unresolved despite the proposal of various surgical procedures. We developed a new polyglycolic acid (PGA) fabric composed of fine diameter fibers to prevent POPF, and macroscopically and microscopically evaluated the effects of applying it to the pancreatic remnant. METHODS: The ventral pancreatic surface was cauterized to create the experimental model of POPF in 33 female Wistar/ST rats. The injured sites were wrapped with nonwoven PGA fabrics of different fiber diameters and porosities in the treated rats; one group of rats remained untreated. Survival, incidence of generalized peritonitis, and microscopic findings around the pancreas were investigated. RESULTS: The PGA fabrics acted as a scaffold for tissue repair and resulted in superior survival. Generalized peritonitis was milder in the PGA treated groups. With the new PGA fabric, abundant fibroblast infiltration and a uniformly-developed, self-organized barrier wall prevented both pancreatic leak and spread of inflammation. CONCLUSION: Application of the newly developed PGA fabric to the pancreatic remnant prevented POPF, and the essential factor for preventing pancreatic leak was the early formation of a self-organized barrier.

Two-layer sheet of gelatin: A new topical hemostatic agent.

BACKGROUND/OBJECTIVE: Uncontrolled surgical bleeding is associated with increased morbidity, mortality, and hospital cost. Topical hemostatic agents available today have problems controlling hemostatic effects; furthermore, their handling is difficult and they are unsafe. METHODS: We devised a new hemostatic agent comprising gelatin sponge and film designed to be applied to the bleeding site, thereby creating a topical hemostatic agent made of gelatin alone. The gelatin was prepared by alkali treatment to eliminate viral activity. Hemostatic effects, surgical handling, and tissue reactions of the materials, namely a two-layer sheet of gelatin, TachoSil, and gelatin sponge, were evaluated using 21 dogs' spleens. RESULTS: The two-layer gelatin sheet and gelatin sponge exhibited superior hemostatic effects (100% hemostasis completed) compared with TachoSil (0-17% hemostasis). The gelatin matrix immediately absorbed blood flowing from wounds and activated the autologous components in the absorbed blood that promoted coagulation at the bleeding site. The two-layer gelatin sheet had the best surgical handling among the evaluated materials. Materials made of gelatin were associated with fewer inflammatory reactions compared with materials of TachoSil. CONCLUSION: The two-layer sheet of gelatin is a useful topical agent because of its superior hemostatic effects and usability, and is associated with a lower risk of transmitting diseases and inflammatory reactions.

Newly Developed Polyglycolic Acid Reinforcement Unified with Sodium Alginate to Prevent Adhesion.

Polyglycolic acid (PGA) mesh fabric is widely used for reinforcing injured tissues during surgeries. However, PGA induces chronic inflammation and adhesion. The purpose of this study is to develop PGA reinforcement "without PGA-induced adhesion." We developed a reinforcement fabric unified with PGA mesh and alginate foam. The antiadhesive effects of sodium alginate foam and calcium alginate foam were evaluated in rats. Sodium alginate foam unified with PGA mesh fabric exhibited strong effects that limit the extent and severity of adhesion, whereas calcium alginate foam unified with PGA mesh was less effective in preventing adhesion. In the sodium alginate group, fibroblasts and collagen fibers around implanted sites were sparse and the material degraded rapidly by macrophage ingestion. Fibroblasts and collagen fibers play a major role in adhesion formation and their excessive proliferation results in postoperative adhesion. Thus, inhibiting their increase is the key in preventing PGA-induced adhesion. The reinforcement that is composed of PGA mesh unified with sodium alginate foam strongly inhibited PGA-induced adhesion and showed excellent handling during surgery and could be easily applied with a one-step procedure.

An attempt to measure the diametric relationship between slow and quick phases of nystagmus.

OBJECTIVE: To investigate whether our original method can precisely evaluate the angle between slow and quick phases of nystagmus (vector angle) and to determine whether vector angle analysis is helpful in differentiating between horizontal nystagmus and mixed nystagmus with horizontal and vertical components. METHODS: We included 20 healthy volunteers, 17 patients with horizontal nystagmus, and 15 patients with mixed nystagmus. Caloric nystagmus was recorded in healthy volunteers; positional nystagmus was recorded in each patient. We extracted the velocity of nystagmus from eye movement of each subject and analysed the vector angle. RESULTS: In caloric nystagmus, the vector angle approached 180 degrees as slow-phase velocity increased, suggesting that our vector angle measurement is more reliable with faster nystagmus. Importantly, in horizontal nystagmus from peripheral vestibular disease, the vector angle similarly approached 180 degrees as slow-phase velocity increased; in contrast, the vector angle in cases of mixed nystagmus from vertebrobasilar insufficiency or spinocerebellar degeneration significantly differed from the angle of caloric nystagmus. CONCLUSIONS: Vector angle analysis using our original algorithm can precisely evaluate the diametric relationship in vestibular nystagmus; it may be helpful in diagnosis of non-peripheral vestibular disorders.

Physical and biological properties of a novel anti-adhesion material made of thermally cross-linked gelatin film: Investigation of the usefulness as anti-adhesion material.

To create more useful, effective and safer anti-adhesion materials, we developed a thermally cross-linked gelatin film. In this study, we examined the physical properties of the film such as the physical strength and the adhesiveness to reveal the handling properties and biological properties, such as the anti-adhesion effect, the influence on cell proliferation, and the cytotoxicity to reveal the anti-adhesion mechanism, especially in comparison with the conventional hyaluronic acid and carboxymethylcellulose film (the conventional film). A tensile test under dry and wet conditions and shearing stress test showed that the gelatin film has significant higher maximum tensile stress and fracture strain than the conventional film. In the study using a rat model of cecum adhesion, the anti-adhesion effect of the gelatin film was significantly superior to that of the conventional film. In the cell proliferation test, the number of fibroblast cells on the gelatin film increased at each time point, while no cell proliferation was observed on the conventional film. Furthermore, in the cytotoxicity test using a colony assay and Live/Dead assay, the extract of the gelatin film had no cytotoxicity, while the extract of the conventional film had cytotoxicity considerably. These results suggest that the gelatin film provides better handling than the conventional film, due to better physical strength and ductility of the film. In addition, the gelatin film has a significantly greater anti-adhesion effect than the conventional film without any cytotoxicity. Therefore, the gelatin film is quite favorable as an anti-adhesion material. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 689-696, 2018.

Constitutive activation of DIA1 (DIAPH1) via C-terminal truncation causes human sensorineural hearing loss.

DIAPH1 encodes human DIA1, a formin protein that elongates unbranched actin. The c.3634+1G>T DIAPH1 mutation causes autosomal dominant nonsyndromic sensorineural hearing loss, DFNA1, characterized by progressive deafness starting in childhood. The mutation occurs near the C-terminus of the diaphanous autoregulatory domain (DAD) of DIA1, which interacts with its N-terminal diaphanous inhibitory domain (DID), and may engender constitutive activation of DIA1. However, the underlying pathogenesis that causes DFNA1 is unclear. We describe a novel patient-derived DIAPH1 mutation (c.3610C>T) in two unrelated families, which results in early termination prior to a basic amino acid motif (RRKR1204-1207) at the DAD C-terminus. The mutant DIA1(R1204X) disrupted the autoinhibitory DID-DAD interaction and was constitutively active. This unscheduled activity caused increased rates of directional actin polymerization movement and induced formation of elongated microvilli. Mice expressing FLAG-tagged DIA1(R1204X) experienced progressive deafness and hair cell loss at the basal turn and had various morphological abnormalities in stereocilia (short, fused, elongated, sparse). Thus, the basic region of the DAD mediates DIA1 autoinhibition; disruption of the DID-DAD interaction and consequent activation of DIA1(R1204X) causes DFNA1.

Prevention of Polyglycolic Acid-Induced Peritoneal Adhesions Using Alginate in a Rat Model.

Postoperative intra-abdominal or intrathoracic adhesions sometimes cause significant morbidity. We have designed three types of alginate-based treatments using strongly cross-linked (SL), weakly cross-linked (WL), and non-cross-linked (NL) alginate with calcium gluconate. In rat experiments, we compared the antiadhesive effects of the three types of alginate-based treatments, fibrin glue treatment (a standard treatment), and no treatment against adhesions caused by polyglycolic acid (PGA) mesh (PGA-induced adhesions). The antiadhesive materials were set on the PGA sheet fixed on the parietal peritoneum of the abdomen. Fifty-six days later, the adhesions were evaluated macroscopically by the adhesion scores and microscopically by hematoxylin-eosin staining and immunostaining. We also tested the fibroblast growth on the surface of the antiadhesive materials in vitro. The antiadhesive effects of WL and NL were superior to the no treatment and fibrin glue treatment. A microscopic evaluation confirmed that the PGA sheet was covered by a peritoneal layer constructed of well-differentiated mesothelial cells, and the inflammation was most improved in the NL and WL. The fibroblast growth was inhibited most on the surfaces of the NL and WL. These results suggest that either the WL or NL treatments are suitable for preventing PGA-induced adhesions compared to SL or the conventional treatment.

Biological properties of a thermally crosslinked gelatin film as a novel anti-adhesive material: Relationship between the biological properties and the extent of thermal crosslinking.

In order to prevent postoperative adhesion and the related complications, a thermally crosslinked gelatin (TCG) film was developed and the basic biological properties were examined, paying special attention to the relationship between these properties and the extent of crosslinking of the film. The gelatin films crosslinked thermally for five different time periods (0, 1, 3, 8, and 14 hours) were developed and the following tests were performed. Regarding the material characterization of the films, the water content, the water solubility, and the enzymatic degradation for collagenase were found to be closely related to the duration of thermal crosslinking. In an in vitro study conducted to examine the cell growth of fibroblasts cultured on the films, the degree of cell growth, except no crosslinked film, was less than that observed in the control group, thus suggesting that such effects of the films on fibroblast cell growth may be related with their anti-adhesive effects. In in vivo tests, the films crosslinked for longer time periods (3, 8, and 14 hours) were retained for longer after being implanted into the abdominal cavity in rats and showed a significant anti-adhesive effect in the rat cecum adhesion models, indicating that the biodegradability and anti-adhesive effects of the TCG films depend on the duration of thermal crosslinking. In order to develop useful and effective anti-adhesive gelatin film, it is very important to optimize duration of the thermal crosslinking.

One-third of vertiginous episodes during the follow-up period are caused by benign paroxysmal positional vertigo in patients with Meniere's disease.

CONCLUSION: In the present study, about one-third of patients with Meniere's disease developed benign paroxysmal positional vertigo (BPPV)-like attacks. Additionally, more than one-third of all vertigo attacks were BPPV-like attacks. Thus, vertigo attacks in patients with Meniere's disease must be carefully treated because the therapy for such vertigo attacks is totally different from the therapy for BPPV. OBJECTIVE: Physicians sometimes encounter patients with previously diagnosed Meniere's disease who develop BPPV attacks during the course of clinical follow-up. In this study, we explored the frequency with which BPPV was involved in all vertiginous episodes among patients with Meniere's disease. METHODS: This retrospective study involved 296 patients with Meniere's disease who visited Kyoto University Hospital. The diagnosis of Meniere's disease was based on the guidelines for the diagnosis of Meniere's disease proposed by the Committee on Hearing and Equilibrium. We judged the cause of vertigo as one of the following five types: (1) definite Meniere's disease attack, (2) suspicious Meniere's disease attack, (3) definite BPPV attack, (4) suspicious BPPV attack, or (5) unknown. RESULTS: In all, 96 patients (32.8%) developed BPPV-like attacks, and 187 vertiginous episodes (37.9%) were caused by BPPV. The lateral semicircular canal was the most frequently involved canal.

In vivo imaging of mouse cochlea by optical coherence tomography.

HYPOTHESIS: Cochlear pathology can be evaluated in living animals using optical coherence tomography (OCT). BACKGROUND: The current imaging methods available for the detailed analysis of cochlear pathology in a clinical setting provide only limited information. Thus, a cochlear imaging modality with high definition is needed for improving the diagnosis of cochlear pathology. OCT has been used in other fields for obtaining high-resolution subsurface images, and its use could potentially be extended to the analysis of cochlear pathogenesis. METHODS: Slc26a4(-/-) mice, which generate endolymphatic hydrops, and their littermates were used in this study. Auditory function was monitored by the auditory brainstem responses (ABR). After the mice were placed under general anesthesia, OCT images of the cochlea were captured. The cochlea was subsequently dissected out and histologically evaluated. Three or 7 days later, the wild-type mice cochleae were visualized again. RESULTS: In ABR assessments, Slc26a4(-/-) mice showed severe hearing loss, while no significant hearing loss was found in Slc26a4(+/-) or Slc26a4(+/+) mice. OCT demonstrated normal morphology in the cochlea of both Slc26a4(+/-) and Slc26a4(+/+) mice, including the location of Reissner's membrane. Meanwhile, in Slc26a4(-/-) mice, obvious dislocation of Reissner's membrane was observed, indicating severe endolymphatic hydrops. These findings in the OCT images were consistent with the histologic results for the cochlear morphology, as observed with hematoxylin and eosin staining. Three or 7 days later, wild-type cochleae were successfully visualized using OCT, and no otitis media or labyrinthitis was observed. CONCLUSION: OCT can be applied in the detection of endolymphatic hydrops in living mice, indicating the potential of OCT for cochlear imaging analyses for clinical use in the near future.