In-process monitoring of a tissue-engineered oral mucosa fabricated on a micropatterned collagen scaffold: use of optical coherence tomography for quality control.

Background: We previously reported a novel technique for fabricating dermo-epidermal junction (DEJ)-like micropatterned collagen scaffolds to manufacture an ex vivo produced oral mucosa equivalent (EVPOME) for clinical translation; however, more biomimetic micropatterns are required to promote oral keratinocyte-based tissue engineering/regenerative medicine. In addition, in-process monitoring for quality control of tissue-engineered products is key to successful clinical outcomes. However, evaluating three-dimensional tissue-engineered constructs such as EVPOME is challenging. This study aimed to update our technique to fabricate a more biomimetic DEJ structure of oral mucosa and to investigate the efficacy of optical coherence tomography (OCT) in combination with deep learning for non-invasive EVPOME monitoring. Methods: A picosecond laser-textured microstructure mimicking DEJ on stainless steel was used as a negative mould to fabricate the micropatterned collagen scaffold. During EVPOME manufacturing, OCT was applied twice to monitor the EVPOME and evaluate its epithelial thickness. Findings: Our moulding system resulted in successful micropattern replication on the curved collagen scaffold. OCT imaging visualised the epithelial layer and the underlying micropatterned scaffold in EVPOME, enabling to non-invasively detect specific defects not found before the histological examination. Additionally, a gradual increase in epithelial thickness was observed over time. Conclusion: These findings demonstrate the feasibility of using a stainless-steel negative mould to create a more biomimetic micropattern on collagen scaffolds and the potential of OCT imaging for quality control in oral keratinocyte-based tissue engineering/regenerative medicine.

HCV genotype 4, 5 and 6: Distribution of viral subtypes and sustained virologic response rates in clinical trials of approved direct-acting antiviral regimens.

Multiple direct-acting antiviral (DAA)-based regimens are now available for all hepatitis C virus (HCV) genotypes (GTs). Because HCV GT 4, 5 and 6 are less common in the United States (US) and worldwide, relatively small numbers of participants with these GTs were evaluated in individual clinical trials. To provide a comprehensive description of subtype diversity and treatment outcomes in clinical trials for these less common GTs, we analysed data from 744 participants with HCV GT4 (n = 573), GT5 (n = 81), or GT6 (n = 90) across 18 clinical trials of DAA regimens. These data are from US New Drug Applications submitted between 2014 and 2017, and our analyses included only approved regimens. Excluding unresolved or mixed subtypes, the distribution of reported GT4 subtypes was 49% 4a, 31% 4d and 16% for one of 14 other subtypes. The distribution of GT6 subtypes was 39% 6a, 27% 6e, 8% 6 L and 23% for one of 11 other subtypes. Across approved regimens, sustained virologic response rates 12 weeks post-treatment (SVR12) for GT 4, 5 and 6 ranged from 91% to 100%, 93% to 97% and 96% to 100%, respectively. SVR12 by GT4 subtype ranged from 96% to 100% for 4a and 81% to 100% for 4d. Virologic failures occurred in GT 4a, 4b, 4d and 4r. For GT6, SVR12 was 100% for all subtypes except 6 L, for which 1 of 7 participants experienced virologic failure. To our knowledge, this is the largest compilation of HCV GT 4, 5 or 6 clinical trial data. These analyses may be useful for clinicians treating HCV GT 4, 5 or 6.

Significance of caveolin-1 and matrix metalloproteinase 14 gene expression in canine mammary tumours.

Canine mammary tumours (CMTs) are the most common neoplasms affecting female dogs. There is an urgent need for molecular biomarkers that can detect early stages of the disease in order to improve accuracy of CMT diagnosis. The aim of this study was to examine whether caveolin-1 (Cav-1) and matrix metalloproteinase 14 (MMP14) are associated with CMT histological malignancy and invasion. Sixty-five benign and malignant CMT samples and six normal canine mammary glands were analysed using quantitative reverse transcription-polymerase chain reaction. Cav-1 and MMP14 genes were highly expressed in CMT tissues compared to normal tissues. Cav-1 especially was overexpressed in malignant and invasive CMT tissues. When a CMT cell line was cultured on fluorescent gelatin-coated coverslips, localisation of Cav-1 was observed at invadopodia-mediated degradation sites of the gelatin matrix. These findings suggest that Cav-1 may be involved in CMT invasion and that the markers may be useful for estimating CMT malignancy.

Whole-genome prediction of fatty acid composition in meat of Japanese Black cattle.

Because fatty acid composition influences the flavor and texture of meat, controlling it is particularly important for cattle breeds such as the Japanese Black, characterized by high meat quality. We evaluated the predictive ability of single-step genomic best linear unbiased prediction (ssGBLUP) in fatty acid composition of Japanese Black cattle by assessing the composition of seven fatty acids in 3088 cattle, of which 952 had genome-wide marker genotypes. All sires of the genotyped animals were genotyped, but their dams were not. Cross-validation was conducted for the 952 animals. The prediction accuracy was higher with ssGBLUP than with best linear unbiased prediction (BLUP) for all traits, and in an empirical investigation, the gain in accuracy of using ssGBLUP over BLUP increased as the deviations in phenotypic values of the animals increased. In addition, the superior accuracy of ssGBLUP tended to be more evident in animals whose maternal grandsire was genotyped than in other animals, although the effect was small.

Haemoglobin wrapped covalently by human serum albumin mutants containing Mn(III) protoporphyrin IX: an O2 complex stable in H2O2 solution.

We describe the synthesis of a haemoglobin (Hb) wrapped covalently by recombinant human serum albumin mutants [HSA(Y161H)] containing Mn(III) protoporphyrin IX (MnPP), the Hb-[HSA(Y161H)-MnPP]3 cluster, highlighting the formation of its O2-complex stable even in H2O2 solution.

An Escherichia coli trap in human serum albumin microtubes.

We describe the template synthesis of human serum albumin microtubes (MTs) and highlight their Escherichia coli (E. coli) trapping capability with extremely high efficiency. The E. coli was loaded into the one-dimensional pore space interior of the tubule. Similar MTs including an Fe3O4 layer also captured E. coli and were manipulated by exposure to a magnetic field.

Genomic prediction in Japanese Black cattle: application of a single-step approach to beef cattle.

The implementation of genomic selection for Japanese Black cattle, known for rich marbling of their meat, is now being explored. Although multiple-step methods are often adopted for dairy cattle, they present shortcomings such as bias and loss of information in addition to operational complexity. These can be avoided using single-step genomic BLUP (ssGBLUP) based on the relationship matrix H, which is constructed from the numerator relationship matrix (A) augmented by the genomic relationship matrix (G). This study assessed the use of ssGBLUP for 3 economically important traits in Japanese Black cattle. Three aspects of ssGBLUP that are important for practical use were examined specifically: the mixing proportions of blending G with A, selection of subsets of genotyped animals used for constructing H, and prediction ability for ungenotyped animals. Different mixing proportions were tested to assess the influence of these proportions on variance component estimation and prediction accuracy. For all traits, the highest or nearly highest accuracy was obtained when the adopted mixing proportion provided heritability closest to that inferred based on A. However, the accuracy did not increase greatly under adjustment of the mixing proportion, thereby suggesting that the influence of the mixing proportion on the accuracy was limited. Genotype data of influential bulls showed a greater contribution to accuracy than that of bulls that were less influential. Genotyping animals with phenotypic records increased the accuracy. It can be prioritized over genotyping bulls that are not influential on the population. These results are expected to present good guides to the future expansion of genotyped populations. Even for animals without genotype data but with genotyped sires, ssGBLUP provided more accurate prediction than BLUP did. For both phenotype and breeding value prediction, ssGBLUP provides more accurate prediction than BLUP, suggesting its usefulness in genomic selection in Japanese Black cattle.

Spatial distributions of floating seaweeds in the East China Sea from late winter to early spring.

Floating seaweeds play an important role as a habitat for many animals accompanying or attaching to them in offshore waters. It was in 2000 that the first report described abundant distributions of floating seaweeds in offshore waters in the East China Sea in spring. Young individuals of the yellowtail Seriola quinqueradiata are captured for aquaculture purposes from floating seaweeds in the East China Sea. Therefore, a sound understanding of the distributions of floating seaweeds in the East China Sea is needed. Detailed information is especially important during the late winter to early spring, which corresponds to the juvenile period of the yellowtail. Thus, field surveys using R/V Tansei-Maru were conducted in the Japanese Exclusive Economic Zone in the East China Sea from late winter to early spring in 2010 and 2011. We obtained positions of the vessel by GPS and transversal distances from the vessel to a raft by visual observation. Distance sampling method (Thomas et al. 2010) was applied to estimation of floating seaweed densities (rafts km-2). Seaweed rafts were also randomly sampled using nets during the research cruises. In the East China Sea, seaweed rafts were distributed mainly on the continental shelf west of the Kuroshio, especially in waters between 26° N and 30° N. Collected rafts consisted of only one species, Sargassum horneri (Turner) C. Agardh. Taking into account surface currents and geographical distribution of S. horneri, it is estimated that these floating seaweeds originated from natural beds along the coast between mid and south China. Considering the approximate travel times, it is suggested that floating patches are colonized by yellowtails early on during their trips, i.e., close to the Chinese coast.

Factors having effect on graft survival in cadaveric renal transplantation.

BACKGROUND: One of the problems of cadaveric renal transplantation is that its graft survival rate is less than that for living renal transplantation. We aim to study relationships between the graft survival of cadaveric renal transplantation patients and various factors. MATERIALS AND METHODS: We retrospectively analyzed 350 cadaveric renal transplantation patients from our institutions from 1983 to 2011. Kaplan-Meier analysis was performed to evaluate graft survival ratios. Using a multivariable Cox regression model, we evaluated the relationship between graft survival and the factors such as age and gender of donor and recipient, body mass index of recipient, duration of hemodialysis, warm ischemic time, and acute rejection (AR), etc. RESULTS: Among 235 males and 115 females, the overall mean age was 41 years. Median follow-up was 15 years (2 to 28 years). The graft survival ratio was 97% at 1 year, 85% at 5 years, and 71% at 10 years. Using the Cox regression model, graft survival was affected by donor age (younger than 60 years; hazard ratio [HR] 1.5; 95% confidence interval (CI) 1.0-2.0; P = .027) and early acute rejection (within 3 months; HR 2.1; CI 1.6-2.8; P < .001). CONCLUSIONS: The graft survival of cadaveric renal transplantation patients is affected by factors of donor age and early AR.

Involvement of peripheral cannabinoid and opioid receptors in ß-caryophyllene-induced antinociception.

BACKGROUND: ß-caryophyllene (BCP) is a common constitute of the essential oils of numerous spice, food plants and major component in Cannabis. The present study investigated the contribution of peripheral cannabinoid (CB) and opioid systems in the antinociception produced by intraplantar (i.pl.) injection of BCP. The interaction between peripheral BCP and morphine was also examined. METHODS: The antinociceptive effect of i.pl. BCP was assayed by the capsaicin tests in mice. Antagonists for CB and opioid receptors, and antisera against ß-endorphin were injected peripherally prior to i.pl. injection of BCP. Morphine in combination with BCP was injected subcutaneously or intrathecally. RESULTS: The i.pl. injection of BCP dose-dependently attenuated capsaicin-induced nociceptive response. The antinociceptive effect produced by BCP was prevented by pretreatment with AM630, a selective CB2 receptor antagonist, but not by AM251, a selective CB1 receptor antagonist. Pretreatment with naloxone, an opioid receptor antagonist, and ß-funaltrexamine, a selective µ-opioid receptor antagonist, reversed the antinociceptive effect of BCP. Pretreatment with naloxone methiodide, a peripherally acting antagonist for opioid receptors and antisera against ß-endorphin, resulted in a significant antagonizing effect on BCP-induced antinociception. Morphine-induced antinociception was increased by a low dose of BCP. The increased effect of morphine in combination with BCP was antagonized significantly by pretreatment with naloxone. CONCLUSIONS: The present results demonstrate that antinociception produced by i.pl. BCP is mediated by activation of CB2 receptors, which stimulates the local release from keratinocytes of the endogenous opioid ß-endorphin. The combined injection of morphine and BCP may be an alternative in treating chemogenic pain.

An analysis of the survival rate after radiotherapy in lung cancer patients with bone metastasis: is there an optimal subgroup to be treated with high-dose radiation therapy?

We investigated the prognostic factors after radiotherapy for bone metastasis from lung cancer while taking the recent findings in the treatment of such cases into consideration. A total of 132 patients with bone metastases from pathologically confirmed lung cancer were evaluated regarding the following potential prognostic factors: treatment for primary site (surgery vs. other), treatment site (spine vs. other), number of bone metastases (solitary vs. multiple), number of metastatic organs (0 vs. 1 vs. ≥2), neurological symptoms (no symptoms vs. numbness vs. paresis), degree of pain (no pain vs. mild pain vs. severe pain), performance status [PS] (0-1 vs. ≥2), biological effective dose [BED] (≥40 Gy vs. <40Gy), time to distant metastasis (≥1 year vs. <1 year), histology (adenocarcinoma vs. others), and use of epidermal growth factor receptor [EGFR]-targeted agents (Yes vs. No). The univariate analysis demonstrated that all factors except for the treatment site were significant. Surgery as treatment for primary site, solitary bone metastasis, no visceral organ metastasis, no symptoms or numbness, no pain, PS<2, BED≥40 Gy, time to distant metastasis ≥ 1year, adenocarcinoma histology, and use of EGFR-targeted agents were correlated with a favorable prognosis. In a multivariate analysis, solitary bone metastasis, PS<2, BED≥40 Gy, adenocarcinoma histology, and the use of EGFR-targeted agents were significantly correlated with a better survival (p = 0.038, 0.006, 0.003, 0.014, and <0.001, respectively). A contingency table to assess the relationship between each variable and the median survival time of the patients according to the administered BED showed that in patients with the time to distant metastasis ≥ 1year and the use of EGFR-targeted agents, the subgroups treated with BED≥40 Gy had a favorable prognosis. Our study suggests that high-dose radiotherapy is associated with a better prognosis in combination with other favorable prognostic factors.

(99m)Tc-mercaptoacetyl triglycine renography to monitor renal transplant function among kidneys from donors after cardiac death.

PURPOSE: Delayed graft function usually occurs after kidney transplantation from donors after cardiac death, It is important to monitor graft function during the anuric period, but there have been few useful tools. Consequently, we evaluated the availability of (99m)-Tc mercaptoacetyltriglycine (MAG3) renography. METHODS: Thirty-two patients underwent renal transplantation from donors after cardiac death between June 2, 2005, and April 14, 2011. One patient was excluded due to an acute rejection episode which developed during the dialysis period. The first (99m)Tc-MAG3 renogram was performed as early as possible after the operation and repeated until the patient was weaned from dialysis. The corrected tubular extraction rate (cTER; mL/min/1.73 m(2)) was calculated; it represents the MAG3 clearance corrected by body surface area. RESULTS: cTER was low immediately after transplantation, but increased gradually until the patient was weaned from dialysis. A significant correlation was observed between early cTER and the period of dialysis-dependence (r = -0.677, P < .001) as well as the short-term best corrected creatinine clearance (r = 0.526, P = .002). CONCLUSION: We observed that graft function can be monitored by routinely performing (99m)Tc-MAG3 renography after transplantation of kidneys from donors after cardiac death.

[Respiratory distress due to rapid expansion of intrapericardial bronchogenic cyst].

A 79-year-old female who had been observed for an intrapericardial cyst for 3 years was admitted due to severe dyspnea and back pain. During 3 years observation, there were no symptoms and no growth of the cyst. Computed tomography scans showed rapid expansion of the cyst compared with an magnetic resonance imaging (MRI) taken 9-days before when she visited our hospital for the 1st time. Urgent complete extirpation of the tumor was successfully done and a histological diagnosis found it was an intrapericardial bronchogenic cyst. This is a 1st report of intrapericardial bronchogenic cyst of which rapid expansion is clearly demonstrated by radiology. It is suggested that complete surgical excision of intrapericardial bronchogenic cysts at the time of diagnosis even if asymptomatic.

Usefulness of a bidirectional e-learning material for explaining surgical anesthesia to cancer patients.

BACKGROUND: We developed an e-learning system, which is based on an interactive animation video that assists anesthesiologists in preanesthetic interviews. MATERIALS AND METHODS: First, the feasibility of the system was investigated in 18 anesthesiologists and 95 volunteers from the general public. Content/quantity, operability, and satisfaction were assessed with a five-point scale. Secondly, a randomized controlled trial was conducted on 211 cancer patients who were scheduled to undergo general anesthesia. They were divided into an e-learning group (n = 106) and a control group (n = 105). The patients in the e-learning group watched the interactive animation before a preanesthetic interview by an anesthesiologist. RESULTS: In 10 of the 11 items for content/quantity, operability, and satisfaction, the average score for both anesthesiologists and volunteers was ≥3.0 in feasibility study. Then, the level of patient comprehension of preoperative rounds and postoperative complications in the e-learning group was significantly higher than that in the control group (mean: 4.4 ± 0.5 versus 4.1 ± 0.7, P = 0.003, and 4.3 ± 0.5 versus 4.2 ± 0.5, P = 0.02); however, no significant difference in anxiety was seen between the two groups. Patient satisfaction in the e-learning group was significantly higher (mean: 4.3 ± 0.5 versus 4.0 ± 0.6, P = 0.002). CONCLUSION: The e-learning system is an effective supplementary tool for preanesthetic interviews in cancer patients.

Prolonged carbon dioxide insufflation under general anesthesia for endoscopic submucosal dissection.

BACKGROUND AND STUDY AIMS: Carbon dioxide (CO (2)) insufflation for endoscopy has been reported to provide superior recovery and is expected to reduce the risk of serious complications, including air embolism and tension pneumothorax, whereas general anesthesia offers some advantages not found under intravenous sedation. Little is known about the effect of prolonged CO (2) insufflation into gastrointestinal tracts on arterial CO (2) tension (PaCO (2)). Here we introduce the use of general anesthesia with CO (2) insufflation for esophagogastroduodenal endoscopic submucosal dissection (ESD). PATIENTS AND METHODS: A prospective observational study was conducted in a university-affiliated hospital. A total of 100 patients were scheduled for esophagogastroduodenal ESD under general anesthesia with CO (2) insufflation, using standardized anesthesia techniques and unchanged ventilatory settings. Arterial blood gas analyses were repeated at predetermined time intervals. RESULTS: Of the initial 100 participants, 94 patients undergoing ESD and four patients undergoing endoscopic mucosal resection completed the study. The median procedure time was 122 minutes (range 29 - 309 minutes). The median baseline PaCO (2) of 28 mmHg increased to a median peak PaCO (2) of 39 mmHg ( P < 0.001), with marked inter-individual variability in the time courses of changes in PaCO (2). The correlation coefficient of PaCO (2) with the procedure time was low (r = 0.194; n = 577, P < 0.0001). FEV (1.0) % (forced expiratory volume in 1 second/forced vital capacity) of < 70 % and esophagoscopy vs. gastroduodenoscopy were relative enhancement factors of PaCO (2). CONCLUSION: Increases of PaCO (2) during esophagogastroduodenal ESD under general anesthesia with CO (2) insufflation remained within acceptable or readily controllable ranges, and are little enhanced by prolongation of the procedure. Esophagogastroduodenal ESD can be performed safely and feasibly with this procedure.

Endotoxin-induced lung injury in α-galactosylceramide-sensitized mice is caused by failure of interleukin-4 production in lung natural killer T cells.

Administration of bacterial lipopolysaccharide (LPS) known as endotoxin into α-galactosylceramide (α-GalCer)-sensitized mice causes severe lung lesions but few hepatic lesions in lethal shock, and interferon (IFN)-γ is suggested to play a pivotal role in preparation of the lung lesions. In order to clarify the mechanism of how α-GalCer sensitization causes lung lesions exclusively in mice, we examined the differential responsiveness of lungs and livers to α-GalCer sensitization. Although lung and liver natural killer T (NK T) cells both produced IFN-γ in response to α-GalCer, IFN-γ signalling was triggered only in the lungs of α-GalCer-sensitized mice. Lung NK T cells did not produce interleukin (IL)-4 in response to α-GalCer and it did not induce the expression of suppressor of cytokine signalling 1 (SOCS1) in the lungs. Conversely, IL-4 produced by liver NK T cells led to the expression of SOCS1 in the livers of the mice. Neutralization of IL-4 reduced SOCS1 expression in the livers and exacerbated LPS-induced hepatic lesions. IL-10 was produced by liver NK T cells but not lung NK T cells. However, IL-10 was produced constitutively by alveolar epithelial cells in normal lung. Lung NK T cells and liver NK T cells might express CD8 and CD4, respectively. Based on the fact that IL-4 inhibited IFN-γ signalling in the livers of α-GalCer-sensitized mice via SOCS1 expression and signal transducer and activator of transcription 1 (STAT-1) activation, no inhibition of the IFN-γ signalling in the lungs caused LPS-induced lung lesions in α-GalCer-sensitized mice. The detailed mechanism of development of the lung lesions in α-GalCer-sensitized mice is discussed.

Metformin attenuates production of nitric oxide in response to lipopolysaccharide by inhibiting MyD88-independent pathway.

Metformin is reported to ameliorate inflammation in diabetic patients. The effect of metformin on lipopolysaccharide-induced nitric oxide production was studied by using RAW 264.7 macrophage-like cells. The action of metformin was analyzed by dividing lipopolysaccharide signaling into the MyD88-dependent and -independent pathways. Metformin significantly reduced the expression of an inducible type of nitric oxide synthase and inhibited lipopolysaccharide-induced nitric oxide production. On the other hand, metformin did not inhibit lipopolysaccharide-induced tumor necrosis factor-alpha production. The expression levels of interferon-beta protein and mRNA, which is a key molecule in MyD88-independent pathway, were significantly inhibited by metformin. Compound C, a specific AMP-activated protein kinase inhibitor, did not affect the inhibitory action of metformin. Metformin was suggested to inhibit lipopolysaccharide-induced nitric oxide production via inhibition of interferon-beta production in MyD88-independent pathway. Metformin might exhibit an anti- inflammatory action on diabetic complications as well as the antidiabetic action.

[Solitary fibrous tumor of the seminal vesicle]. / Solitärer fibröser Tumor der Samenblase.

A 56-year-old man presented to our hospital with a pelvic mass. The tumor was diagnosed to occur from right seminal vesicle and to be a benign solitary fibrous tumor by transrectal tumor biopsy. The tumor enlarged during follow up, and he under-went resection of the tumor.