Effects of paclitaxel before vitrification on the nuclear maturation and development of immature porcine oocytes.

BACKGROUND: Cryopreservation of porcine oocytes is difficult compared with other species and immature oocytes particularly so compared to the meiotic stage. OBJECTIVE: To evaluate the efficacy of a pretreatment with 1 micromole per L paclitaxel (PTX, 30 min exposure) before vitrification to promote the maturation of porcine immature oocytes. MATERIALS AND METHODS: Cumulus cell-enclosed oocytes (COs) aspirated from porcine ovaries were divided into three groups: i) non-pretreated with PTX and non-vitrified group (control group); ii) pretreated with PTX and vitrified group (PTX-V group); and iii) non-pretreated with PTX and vitrified group (nPTX-V group). RESULTS: The nuclear maturation rate up to the preovulatory stage was significantly lower (P < 0.05) in the nPTX-V group than in the control group, but was similar in the PTX-V and control groups. No significant differences were observed in viability assessed by a normal CO morphology and the embryonic development of oocytes activated by the parthenogenetic stimulation between the PTX-V and control groups, but not the non-PTX-V group. CONCLUSION: PTX may promote the maturation of vitrified porcine immature oocytes. Doi.org/10.54680/fr23510110812.

Central odontogenic fibroma with amyloid: a diagnostically challenging case.

Odontogenic fibroma is a rare benign mesenchymal odontogenic tumor, with its histological diversity possibly posing diagnostic challenges. A case of the amyloid variant of central odontogenic fibroma, with epithelial cells in perineural and intraneural locations, is reported herein. The 46-year-old female patient had experienced discomfort related to her anterior right hard palate for approximately 25 years. Clinical examination revealed a depression in the anterior hard palate, and radiographic examination showed a well-defined radiolucent lesion with root resorption of the adjacent teeth. Histologically, the well-circumscribed tumor was composed of hypocellular collagenous connective tissue with small islands of odontogenic epithelium. In addition, the juxta-epithelial deposition of amyloid globules without calcification and epithelial cells in perineural and intraneural locations were observed, which posed a diagnostic challenge in differentiating the lesion from the non-calcifying variant of calcifying epithelial odontogenic tumor and sclerosing odontogenic carcinoma. However, on the basis of the clinical and radiographic findings, which were suggestive of a benign and slowly progressive process given the corticated, unilocular radiolucency, the considerable root resorption, and the long history of this finding in an otherwise healthy patient, the final diagnosis was amyloid variant of central odontogenic fibroma. Increased recognition of this variant of odontogenic fibroma and its differentiation from other more aggressive lesions could help the clinician to avoid overdiagnosis and overtreatment.

QSAR model to predict Kp,uu,brain with a small dataset, incorporating predicted values of related parameter.

The unbound brain-to-plasma concentration ratio (Kp,uu,brain) is a parameter that indicates the extent of central nervous system penetration. Pharmaceutical companies build prediction models because many experiments are required to obtain Kp,uu,brain. However, the lack of data hinders the design of an accurate prediction model. To construct a quantitative structure-activity relationship (QSAR) model with a small dataset of Kp,uu,brain, we investigated whether the prediction accuracy could be improved by incorporating software-predicted brain penetration-related parameters (BPrPs) as explanatory variables for pharmacokinetic parameter prediction. We collected 88 compounds with experimental Kp,uu,brain from various official publications. Random forest was used as the machine learning model. First, we developed prediction models using only structural descriptors. Second, we verified the predictive accuracy of each model with the predicted values of BPrPs incorporated in various combinations. Third, the Kp,uu,brain of the in-house compounds was predicted and compared with the experimental values. The prediction accuracy was improved using five-fold cross-validation (RMSE = 0.455, r2 = 0.726) by incorporating BPrPs. Additionally, this model was verified using an external in-house dataset. The result suggested that using BPrPs as explanatory variables improve the prediction accuracy of the Kp,uu,brain QSAR model when the available number of datasets is small.

A case of nivolumab-induced acute-onset type 1 diabetes mellitus in melanoma.

Nivolumab, an anti-PD-1 antibody, is now considered an important therapeutic agent in several advanced malignancies. However, immune-related adverse events such as endocrinopathies have been reported with its use. Thyroid disorder and isolated adrenocorticotropic hormone deficiency have frequently been reported as nivolumab-induced immune-related adverse events. Another endocrinopathy is nivolumab-induced type 1 diabetes mellitus (t1dm), described as diabetes mellitus with rapid onset and complete insulin insufficiency, at times leading to fulminant t1dm. We report the case of a 68-year-old woman who developed pancreatic islet-related autoantibody-negative t1dm, possibly induced by nivolumab, under continuous glucocorticoid administration. She was treated with nivolumab for advanced malignant melanoma, concomitant with 10 mg prednisolone daily for thrombophlebitis tapered to 5 mg after 13 courses of nivolumab therapy. At approximately the 27th course of nivolumab therapy, she showed elevated plasma glucose levels despite preserved insulin secretion. A month later, she developed diabetic ketoacidosis. Her insulin secretion decreased and finally was exhausted. She was diagnosed with acute-onset rather than fulminant t1dm because of a rapidly progressive course to diabetic ketoacidosis during just more than 1 week. She is currently receiving insulin replacement. There has been no recurrence of the melanoma. Thus, nivolumab might induce autoimmune diabetes mellitus, with patients having t1dm-sensitive human leucocyte antigen being more susceptible even when receiving glucocorticoids. Physicians should be aware that nivolumab could potentially induce t1dm as a critical immune-related adverse event.

Detection of in-stent protrusion (ISP) by intravascular ultrasound during carotid stenting: Usefulness of stent-in-stent placement for ISP.

OBJECTIVES: As in-stent protrusion (ISP) during carotid artery stenting (CAS) may cause postoperative embolism, ISP detection is important. Intravascular ultrasound examination (IVUS) is useful for ISP detection because the blood vessel cross-section can be drawn as a tomogram from the lumen. Our objective was to clarify the occurrence of ISP during CAS using IVUS and relevant factors, and to report the usefulness of stent-in-stent placement when treating ISP. METHODS: In 142 consecutive patients (128 men, average age 71.7 years; 69 symptomatic) who underwent CAS using dual protection and the blood aspiration method, and subsequent IVUS after stent placement were included. The outcome of CAS, and the occurrence rate of ISP and related factors (plaque characteristics, stent design, intraoperative debris capture rate and postoperative diffusion-weighted imaging (DWI) positive rate) were examined. RESULTS: All CAS procedures were successful and no major adverse events (MAEs) were observed at 30 days. ISP was found in 12% (17/142), and stent-in-stent placement was performed in all cases. Vulnerable plaques were observed in 12 of 17 ISP cases (71%). A closed stent was used in 13 of 17 ISP cases (71%). The intraoperative debris capture rate was 100%, and no neurological symptoms were observed in any patients. A significant increase in ISP susceptibility was related to vulnerable plaques and the intraoperative debris capture rate. CONCLUSIONS: Vulnerable plaques and debris capture were significantly correlated with ISP occurrence. In all ISP cases, stent-in-stent placement was performed and good results were obtained. KEY POINTS: • ISP detection during CAS using IVUS is important. • ISP-positive patients were correlated with NASCET ≥ 80%, vulnerable plaques and stent length. • Adequate additional treatment of stent in stenting under reliable protection against ISP-positive patients achieved low perioperative complications.

Mild to Moderate Intrapulmonary Shunting in Pediatric Liver Transplantation: Is Screening Necessary?

BACKGROUND: Despite reported associations between intrapulmonary vascular shunting (IPVS) and morbidity and mortality in pediatric liver transplantation (LT), there are no guidelines for screening. OBJECTIVE: To investigate IPVS before and after pediatric LT. METHODS: Retrospective records review of all pediatric LT (n = 370) from 2005 to 2015 at a single institute in Japan. All children with cirrhosis and clinical suspicion of IPVS without cardiac or pulmonary conditions were included. 99mTechnetium labelled macroaggregated albumin (99mTcMAA) scans were performed before and after LT. The severity of IPVS was graded using shunt ratios. RESULTS: Twenty-four children fulfilled inclusion criteria and underwent Tc99MAA scans. All revealed mild (<20%) to moderate (20%-40%) grades of IPVS. Following LT, the mean shunt ratio regressed from 20.69 ± 6.26% to 15.1 ± 3.4% (P = .06). The median (range) follow-up was 17 (4-85) months. Mortality was zero. The incidence of portal vein thrombosis (4.2%) biliary strictures (12.5%) and graft loss (4.1%) in the study group was not statistically significant compared to the remainder of the 370 transplants (3.2%, 9.4% and 3%, respectively). Sub-group analysis revealed hepatopulmonary syndrome (HPS) in 2 out of 24 children. The mean shunt ratios before and after LT were 39.2 ± 0.77% and 16.2 ± 8.5%, respectively (P = .08). There was 1 complication (intra-abdominal abscess). CONCLUSIONS: HPS is less likely in mild to moderate IPVS. LT may achieve comparable results when performed in the presence of mild to moderate IPVS.

Electronic Structure of Ce-Doped and -Undoped Nd_{2}CuO_{4} Superconducting Thin Films Studied by Hard X-Ray Photoemission and Soft X-Ray Absorption Spectroscopy.

In order to realize superconductivity in cuprates with the T^{'}-type structure, not only chemical substitution (Ce doping) but also postgrowth reduction annealing is necessary. In the case of thin films, however, well-designed reduction annealing alone without Ce doping can induce superconductivity in the T^{'}-type cuprates. In order to unveil the origin of superconductivity in the Ce-undoped T^{'}-type cuprates, we have performed bulk-sensitive hard x-ray photoemission and soft x-ray absorption spectroscopy on superconducting and nonsuperconducting Nd_{2-x}Ce_{x}CuO_{4} (x=0, 0.15, and 0.19) thin films. By postgrowth annealing, core-level spectra exhibited dramatic changes, which we attributed to the enhancement of core-hole screening in the CuO_{2} plane and the shift of chemical potential along with changes in the band filling. The result suggests that the superconducting Nd_{2}CuO_{4} film is doped with electrons despite the absence of the Ce substitution.

Imaging Differences between Neuromyelitis Optica Spectrum Disorders and Multiple Sclerosis: A Multi-Institutional Study in Japan.

BACKGROUND AND PURPOSE: Both clinical and imaging criteria must be met to diagnose neuromyelitis optica spectrum disorders and multiple sclerosis. However, neuromyelitis optica spectrum disorders are often misdiagnosed as MS because of an overlap in MR imaging features. The purpose of this study was to confirm imaging differences between neuromyelitis optica spectrum disorders and MS with visually detailed quantitative analyses of large-sample data. MATERIALS AND METHODS: We retrospectively examined 89 consecutive patients with neuromyelitis optica spectrum disorders (median age, 51 years; range, 16-85 years; females, 77; aquaporin 4 immunoglobulin G-positive, 93%) and 89 with MS (median age, 36 years; range, 18-67 years; females, 68; relapsing-remitting MS, 89%; primary-progressive MS, 7%; secondary-progressive MS, 2%) from 9 institutions across Japan (April 2008 to December 2012). Two neuroradiologists visually evaluated the number, location, and size of all lesions using the Mann-Whitney U test or the Fisher exact test. RESULTS: We enrolled 79 patients with neuromyelitis optica spectrum disorders and 87 with MS for brain analysis, 57 with neuromyelitis optica spectrum disorders and 55 with MS for spinal cord analysis, and 42 with neuromyelitis optica spectrum disorders and 14 with MS for optic nerve analysis. We identified 911 brain lesions in neuromyelitis optica spectrum disorders, 1659 brain lesions in MS, 86 spinal cord lesions in neuromyelitis optica spectrum disorders, and 102 spinal cord lesions in MS. The frequencies of periventricular white matter and deep white matter lesions were 17% and 68% in neuromyelitis optica spectrum disorders versus 41% and 42% in MS, respectively (location of brain lesions, P < .001). We found a significant difference in the distribution of spinal cord lesions between these 2 diseases (P = .024): More thoracic lesions than cervical lesions were present in neuromyelitis optica spectrum disorders (cervical versus thoracic, 29% versus 71%), whereas they were equally distributed in MS (46% versus 54%). Furthermore, thoracic lesions were significantly longer than cervical lesions in neuromyelitis optica spectrum disorders (P = .001), but not in MS (P = .80). CONCLUSIONS: Visually detailed quantitative analyses confirmed imaging differences, especially in brain and spinal cord lesions, between neuromyelitis optica spectrum disorders and MS. These observations may have clinical implications.

Postoperative laryngeal morbidity and intubating conditions using the McGRATH™ MAC videolaryngoscope with or without neuromuscular blockade: a randomised, double-blind, non-inferiority trial.

Tracheal intubation without neuromuscular blockade may be associated with worse intubating conditions and increased laryngeal morbidity. We hypothesised that tracheal intubation using the McGRATH™ MAC videolaryngoscope would not increase postoperative hoarseness, even without neuromuscular blockade. In this prospective, randomised, parallel-group, double-blind, non-inferiority trial, 248 patients were randomly assigned to tracheal intubation with or without neuromuscular blockade using rocuronium. Hoarseness and sore throat were evaluated at 24 h and 48 h postoperatively. The primary outcome was the incidence of hoarseness at 48 h postoperatively with a pre-defined non-inferiority margin of 10%. Hoarseness at 48 h did not differ between the non-paralysed group and the paralysed group (8.1% vs. 13.6%; absolute difference: -5.4%; 95%CI: -13.3 to 2.4). Also, no significant differences were found between the two groups for hoarseness at 24 h (22.8% vs. 27.1%) or for sore throat at 24 h (12.2% vs. 9.3%) and 48 h postoperatively (1.6% vs. 0.8%). Although more patients in the non-paralysed group showed an adducted position of the vocal cords (29.3% vs. 0%), there were no significant group differences in the ease of laryngoscopy (96.7% vs. 98.3%), Cormack grade laryngeal view 1 (97.6% vs. 96.6%) or first-pass success rate (100% vs. 100%). We conclude that when using the McGRATH MAC videolaryngoscope for tracheal intubation, the incidence of postoperative hoarseness was not inferior if neuromuscular blockade was avoided.

Six National University Consortium in Liver Transplant Professionals Training (SNUC-LT) Program in Japan.

BACKGROUND: There has been no public structured training program for transplant surgeons in Japan. However, such a program is crucial for optimizing liver transplant surgery and training young professionals in liver transplant surgery. A comprehensive training program was recently developed and the underlying concepts, structure and curriculum, and results of this program are described here. METHODS: We developed a 3-year training program in 2014 called the Six National University Consortium in Liver Transplant Professionals Training (SNUC-LT) program supported by the Ministry of Education, Culture, Sports, Science, and Technology. This program is based on strong cooperation among 6 national universities (Kumamoto, Okayama, Nagasaki, Kanazawa, Niigata, and Chiba Universities). The program includes various courses to help trainees learn transplant theory and practice as well as to teach surgical skills required to safely perform transplant surgery. RESULTS: Three trainees completed the specially designed 3-year curriculum. They attended lectures on transplant theory for an average of 59 hours and participated in an average of 44 liver transplant surgeries and 51 liver resections for transplant practice. Trainees from low-volume centers had sufficient opportunities to attend operations in high-volume centers because of the cooperative agreement among the universities. After finishing the program, the trainees were certified as talent-proven liver transplant surgeons. CONCLUSIONS: The SNUC-LT program is the first national program in Japan to have strong professional support. Our multicenter program enables young surgeons to have more abundant knowledge, more extensive experience, better surgical skills, and smoother communication skills in the field of liver transplantation.

Effects of biopolyimide molecular design on their silica hybrids thermo-mechanical, optical and electrical properties.

Polymers, derived from bio-derived resources, have gained considerable momentum because of a lower dependence over conventional fossil-based resources without compromising the materials' thermo-mechanical properties. Unique characteristics of organic and inorganic materials can be incorporated by a combination of both to obtain hybrid materials. We have recently developed a series of transparent biopolyimides (BPI) from a biologically derived exotic amino acid, 4-aminocinnamic acid (4ACA) to yield 4-amino truxillic ester (4ATA ester) derivatives. In the present research, the polyimide-precursor was subjected to sol-gel polycondensation reactions with silicon-alkoxide followed by annealing under vacuo to yield a biopolyimide-silica hybrid. The biopolyimide structures (4ATA acid/ester) and their silica hybrids thermo-mechanical, electrical and optical performance were evaluated. It was found that the biopolyimide with 4ATA(ester) yields thermo-mechanically robust films with very high electrical stability as well as optical transparency, plausibly due to the uniform dispersion of the silica particles in the biopolyimide matrix.

Liver Transplantation for Mitochondrial Respiratory Chain Disorder: A Single-Center Experience and Excellent Marker of Differential Diagnosis.

Mitochondrial respiratory chain disorder (MRCD) can cause liver failure requiring liver transplantation (LT), although it is often difficult to diagnose before LT. From 2005 to 2016, 9 MRCD patients with the median age at LT of 6 months underwent LT in our institute. Their clinical courses were retrospectively reviewed and the laboratory parameters were compared between the MRCD patients and 10 patients with acute liver failure unrelated to MRCD (non-MRCD). Five patients had extrahepatic manifestations, including developmental disorders in 3 and failure to thrive in 3, before LT. Only 3 patients (33.3%) were diagnosed before LT. Between MRCD and non-MRCD, lactate was significantly high and lactate-to-pyruvate ratio (L/P ratio) tended to be higher in MRCD. From the receiver operating characteristic curve, the optimal cutoff value of lactate was 50.0 mg/dL and that of L/P ratio was 23.2. Patient survival rate of MRCD was 77.8%, although 2 patients with mitochondrial depletion syndrome suffered from de novo pulmonary hypertension after LT. Our experiences showed the difficulty of preoperative diagnosis, and preoperative extrahepatic manifestations did not always mean poor outcome. Our study showed that lactate value and L/P ratio can be excellent predictors of MRCD.

Predicting skeletal muscle mass from dual-energy X-ray absorptiometry in Japanese prepubertal children.

BACKGROUND/OBJECTIVE: We aimed to develop regression-based prediction equations for estimating total and regional skeletal muscle mass (SMM) from measurements of lean soft tissue mass (LSTM) using dual-energy X-ray absorptiometry (DXA) and investigate the validity of these equations. SUBJECTS/METHODS: In total, 144 healthy Japanese prepubertal children aged 6-12 years were divided into 2 groups: the model development group (62 boys and 38 girls) and the validation group (26 boys and 18 girls). Contiguous MRI images with a 1-cm slice thickness were obtained from the first cervical vertebra to the ankle joints as reference data. The SMM was calculated from the summation of the digitized cross-sectional areas. Total and regional LSTM was measured using DXA. RESULTS: Strong significant correlations were observed between the site-matched SMM (total, arms, trunk and legs) measured by MRI and the LSTM obtained by DXA in the model development group for both boys and girls (R2adj=0.86-0.97, P<0.01, standard error of the estimate (SEE)=0.08-0.44 kg). When these SMM prediction equations were applied to the validation group, the measured total (boys 9.47±2.21 kg; girls 8.18±2.62 kg) and regional SMM were very similar to the predicted values for both boys (total SMM 9.40±2.39 kg) and girls (total SMM 8.17±2.57 kg). The results of the Bland-Altman analysis for the validation group did not indicate any bias for either boys or girls with the exception of the arm region for the girls. CONCLUSIONS: These results suggest that the DXA-derived prediction equations are precise and accurate for the estimation of total and regional SMM in Japanese prepubertal boys and girls.

A three-dimensional cell culture system as an in vitro canine mammary carcinoma model for the expression of connective tissue modulators.

In this study, derived complex carcinoma (CC) and simple carcinoma (SC) cell lines were established and cultured under two-dimensional (2D) and three-dimensional (3D) conditions. The 3D was performed in six-well AlgiMatrix™ (LifeTechnologies®, Carlsbad, CA, USA) scaffolds, resulting in spheroids sized 50-125 µm for CC and 175-200 µm for SC. Cell viability was demonstrated up to 14 days for both models. Epidermal growth factor receptor (EGFR) was expressed in CC and SC in both systems. However, higher mRNA and protein levels were observed in SC 2D and 3D systems when compared with CC (P < 0.005). The connective tissue modulators, metalloproteinases-1, -2, -9 and -13 (MMPs), relaxin receptors 1 and 2 (RXR1 and RXR2) and E-cadherin (CDH1) were quantitated. All were upregulated similarly when canine mammary tumour (CMT)-derived cell lines were cultured under 3D AlgiMatrix, except CDH1 that was downregulated (P < 0.005). These results are promising towards the used of 3D system to increase a high throughput in vitro canine tumour model.

Adhesion patterning by a novel air-lock technique enables localization and in-situ real-time imaging of reprogramming events in one-to-one electrofused hybrids.

Although fusion of somatic cells with embryonic stem (ES) cells has been shown to induce reprogramming, single-cell level details of the transitory phenotypic changes that occur during fusion-based reprogramming are still lacking. Our group previously reported on the technique of one-to-one electrofusion via micro-slits in a microfluidic platform. In this study, we focused on developing a novel air-lock patterning technique for creating localized adhesion zones around the micro-slits for cell localization and real-time imaging of post fusion events with a single-cell resolution. Mouse embryonic fibroblasts (MEF) were fused individually with mouse ES cells using a polydimethylsiloxane (PDMS) fusion chip consisting of two feeder channels with a separating wall containing an array of micro-slits (slit width ∼3 µm) at a regular spacing. ES cells and MEFs were introduced separately into the channels, juxtaposed on the micro-slits by dielectrophoresis and fused one-to-one by a pulse voltage. To localize fused cells for on-chip culture and time-lapse microscopy, we implemented a two-step approach of air-lock bovine serum albumin patterning and Matrigel coating to create localized adhesion areas around the micro-slits. As a result of time-lapse imaging, we could determine that cell division occurs within 24 h after fusion, much earlier than the 2-3 days reported by earlier studies. Remarkably, Oct4-GFP (Green Fluorescent Protein) was confirmed after 25 h of fusion and thereafter stably expressed by daughter cells of fused cells. Thus, integrated into our high-yield electrofusion platform, the technique of air-lock assisted adhesion patterning enables a single-cell level tracking of fused cells to highlight cell-level dynamics during fusion-based reprogramming.

Functional outcome and quality of life after traumatic total brachial plexus injury treated by nerve transfer or single/double free muscle transfers: a comparative study.

AIMS: Between 2002 and 2011, 81 patients with a traumatic total brachial plexus injury underwent reconstruction by double free muscle transfer (DFMT, 47 cases), single muscle transfer (SMT, 16 cases) or nerve transfers (NT, 18 cases). METHODS: They were evaluated for functional outcome and quality of life (QoL) using the Disability of Arm, Shoulder and Hand questionnaire, both pre- and post-operatively. The three groups were compared and followed-up for at least 24 months. RESULTS: The mean shoulder abduction and flexion were comparable in all groups, but external rotation was significantly better in the DFMT group as were range and quantitative power of elbow flexion. Patients who had undergone DFMT had reasonable total active finger movement and hook grip strength. All groups showed improvement in function at a level greater than a minimum clinically important difference. The DFMT group showed the greatest improvement. DISCUSSION: Patients in the DFMT group had a better functional outcome and QoL recovery than those in the NT and SMT groups. TAKE HOME MESSAGE: Double free muscle transfer procedure is capable of restoring maximum function in patients of total brachial plexus palsy.