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Kidney fibrosis is closely associated with the progression of chronic kidney disease (CKD). Furthermore, copper-containing secretory amine oxidases, such as lysyl oxidase (LOX) and LOX-like 1-4 (LOXL1-4), play pivotal roles in the regulation of extracellular components and facilitate fibrosis. In this study, we investigated the regulation of LOX enzymes in human tubular epithelial HK2 cells to help clarify the role of LOX enzymes in kidney fibrosis. Among 5 LOX enzymes, LOXL2 expression is abundantly expressed in HK2 cells. LOX enzymes inhibitor, β-aminopropionitrile, suppressed transforming growth factor-β1 (TGF-β1)-promoted epithelial-to-mesenchymal transition processes in HK2 cells, indicating that LOX enzymes are involved in TGF-β1-mediated fibrotic processes. Recent studies suggest that LOX enzymes are secreted into the extracellular spaces by exosomes and promote fibrotic processes. Similar to the previous reports, we observed that exosomes secreted from HK2 cells carry LOXL2 into the extracellular spaces. Furthermore, we determined that N-glycosylation on the asparagine residues plays a key role in LOXL2 secretion. Amino acid mutations in three asparagine residues, which can be glycosylated, suppressed the secretion of mutated LOXL2. Moreover, N-acetylglucosaminyltransferase 5, an enzyme used for the biosynthesis of β1,6N-acetylglucosamine-branched N-glycans, participated in LOXL2 secretion, and the N-glycosylation inhibitor, glucosamine hydrochloride (GS), inhibited TGF-β1-mediated LOXL2 secretion and fibrotic processes. Overall, TGF-β1 promotes LOXL2 secretion and may participate in kidney fibrosis. Our results provide novel insight into the antifibrotic properties of GS that contribute to the inhibition of CKD progression., (© 2023 Wiley Periodicals LLC.)

Lysyl oxidase (LOX) is a copper-containing enzyme and its overexpression in tumor tissues promote tumor metastasis through the crosslinking of extracellular matrix. Our previous report demonstrated that LOX expression is significantly increased in human leukemic THP-1 cell-derived M2-like macrophages, and histone modification plays a key role in its induction. However, the rigorous mechanism of LOX regulation remains unclear. In this study, we investigated the role of functional transcription factors, hypoxia-inducible factor 1α (HIF1α), signal transducer and activator of transcription 3 (STAT3) and forkhead box O1 (FOXO1) in LOX regulation in M2-like macrophages. HIF1α expression was significantly increased in M2-like macrophages, and HIF1α inhibitor, TX402, suppressed LOX induction. The significant STAT3 activation was also observed in M2-like macrophages. Additionally, LOX induction was canceled in the presence of STAT3 inhibitor, S3I-201, suggesting that HIF1α and STAT3 pathways play a critical role in LOX induction. On the other hand, our ChIP results clearly indicated that the enrichment of FOXO1 within the lox promoter region was dramatically decreased in M2-like macrophages. In this context, knockdown of FOXO1 further enhanced LOX induction. LOX induction and HIF1α binding to the lox promoter region were suppressed in FOXO1-overexpressed cells, suggesting that the FOXO1 binding to the lox promoter region counteracted HIF1α binding to that region. Overall, the present data suggested that both of HIF1α and STAT3 were required for LOX induction in M2-like macrophages, and loss of FOXO1 within the lox promoter region facilitated HIF1α binding to that region which promoted LOX induction., (© 2021 Wiley Periodicals LLC.)

Although photoallergens require UV energy for antigen formation, the subsequent immune response is considered to be the same as in ordinary skin sensitization. Therefore, in vitro tests for skin sensitization should also be applicable for photoallergy testing. In this study, we examined whether activation of the Keap1 (Kelch-like ECH-associated protein 1)-Nrf2 (nuclear factor-erythroid 2-related factor 2)-ARE (antioxidant response element) pathway could be used to assess the photoallergenic potential of chemicals, using the reporter cell line AREc32 or KeratinoSens(TM) . First, we identified an appropriate UVA irradiation dose [5 J cm(-2) irradiation in phosphate-buffered saline (PBS)] by investigating the effect of UV irradiation on ARE-dependent gene induction using untreated or 6-methylcoumarin (6-MC)-treated cells. Irradiation of well-known photoallergens under this condition increased ARE-dependent gene expression by more than 50% compared with both vehicle and non-irradiated controls. When the cut-off value for detecting photoallergens was set at 50% induction, the accuracy of predicting photoallergenic/phototoxic chemicals was 70% in AREc32 cells and 67% in KeratinoSens(TM) cells, and the specificity was 100% in each case. We designate these assays as a photo-ARE assay and photo-KeratinoSens(TM) , respectively. Our results suggest that activation of the Keap1-Nrf2-ARE pathway is an effective biomarker for evaluating both photoallergenic and phototoxic potentials. Either of the above tests might be a useful component of a battery of in vitro tests/in silico methods for predicting the photoallergenicity and phototoxicity of chemicals. Copyright © 2015 John Wiley & Sons, Ltd., (Copyright © 2015 John Wiley & Sons, Ltd.)

An in silico method for predicting percutaneous absorption of cosmetic ingredients was developed by using artificial neural network (ANN) analysis to predict the human skin permeability coefficient (log Kp), taking account of the physicochemical properties of the vehicle, and the apparent diffusion coefficient (log D). Molecular weight and octanol-water partition coefficient (log P) of chemicals, and log P of the vehicles, were used as molecular descriptors for predicting log Kp and log D of 359 samples, for which literature values of either or both of log Kp and log D were available. Adaptivity of the ANN model was evaluated in comparison with a multiple linear regression model (MLR) by calculating the root-mean-square (RMS) errors. Accuracy and robustness were confirmed by 10-fold cross-validation. The predictive RMS errors of the ANN model were smaller than those of the MLR model (log Kp; 0.675 vs 0.887, log D; 0.553 vs 0.658), indicating superior performance. The predictive RMS errors for log Kp and log D with the ANN model after 10-fold cross-validation analysis were 0.723 and 0.606, respectively. Moreover, we estimated the cumulative amounts of chemicals permeated into the skin during 24 hr (Q24hr) from the values of log Kp and log D by applying Fick's law of diffusion. Our results suggest that this newly established ANN analysis method, taking account of the property of the vehicle, could contribute to non-animal risk assessment of cosmetic ingredients by providing a tool for calculating Q24hr, which is required for evaluating the margin of safety.

The sensitizing potential of chemicals is usually identified and characterized using in vivo methods such as the murine local lymph node assay (LLNA). Due to regulatory constraints and ethical concerns, alternatives to animal testing are needed to predict the skin sensitization potential of chemicals. For this purpose, an integrated evaluation system employing multiple in vitro and in silico parameters that reflect different aspects of the sensitization process seems promising. We previously reported that LLNA thresholds could be well predicted by using an artificial neural network (ANN) model, designated iSENS ver. 2 (integrating in vitro sensitization tests version 2), to analyze data obtained from in vitro tests focused on different aspects of skin sensitization. Here, we examined whether LLNA thresholds could be predicted by ANN using in silico-calculated descriptors of the three-dimensional structures of chemicals. We obtained a good correlation between predicted LLNA thresholds and reported values. Furthermore, combining the results of the in vitro (iSENS ver. 2) and in silico models reduced the number of chemicals for which the potency category was under-estimated. In conclusion, the ANN model using in silico parameters was shown to be have useful predictive performance. Further, our results indicate that the combination of this model with a predictive model using in vitro data represents a promising approach for integrated risk assessment of skin sensitization potential of chemicals.

Habitat connectivity is considered to have an important role on the persistence of populations in the face of habitat fragmentation, in particular, for species with conservation concern. However, it can also impose indirect negative effects on native species through the spread of invasive species. Here, we investigated direct and indirect effects of habitat connectivity on populations of invasive bullfrogs and native wrinkled frogs and how these effects are modified by the presence of common carp, a resident shared predator, in a farm pond system in Japan. The distribution pattern analysis using a hierarchical Bayesian modelling indicated that bullfrogs had negative effects on wrinkled frogs, and that these negative effects were enhanced with increasing habitat connectivity owing to the metapopulation structure of bullfrogs. The analysis also suggested that common carp mitigated these impacts, presumably owing to a top-down trophic cascade through preferential predation on bullfrog tadpoles. These presumed interspecific interactions were supported by evidence from laboratory experiments, i.e. predation by carp was more intense on bullfrog tadpoles than on wrinkled frog tadpoles owing to the difference in refuge use. Our results indicate that metacommunity perspectives could provide useful insights for establishing effective management strategies of invasive species living in patchy habitats., (© 2014 The Author(s) Published by the Royal Society. All rights reserved.)

The sensitizing potential of chemicals is usually identified and characterized using in vivo methods such as the murine local lymph node assay (LLNA). Due to regulatory constraints and ethical concerns, alternatives to animal testing are needed to predict skin sensitization potential of chemicals. For this purpose, combined evaluation using multiple in vitro and in silico parameters that reflect different aspects of the sensitization process seems promising. We previously reported that LLNA thresholds could be well predicted by using an artificial neural network (ANN) model, designated iSENS ver.1 (integrating in vitro sensitization tests version 1), to analyze data obtained from two in vitro tests: the human Cell Line Activation Test (h-CLAT) and the SH test. Here, we present a more advanced ANN model, iSENS ver.2, which additionally utilizes the results of antioxidant response element (ARE) assay and the octanol-water partition coefficient (LogP, reflecting lipid solubility and skin absorption). We found a good correlation between predicted LLNA thresholds calculated by iSENS ver.2 and reported values. The predictive performance of iSENS ver.2 was superior to that of iSENS ver.1. We conclude that ANN analysis of data from multiple in vitro assays is a useful approach for risk assessment of chemicals for skin sensitization., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background/aims: The objective of this study was to determine whether cerebral arterial vasoconstriction occurs in relation to postprandial splanchnic blood pooling in cirrhotic patients., Methods: The pulsatility and the resistive indexes and blood flow in the middle cerebral artery were measured by magnetic resonance imaging in 21 cirrhotics and 14 controls. These measurements were repeated 30 min after ingestion of a 400 kcal liquid meal or placebo. Seven controls and 14 patients received the meal, and seven controls and seven patients received placebo., Results: In the fasting conditions, cirrhotics had a greater pulsatility index (0.81 +/- 0.10 vs. 0.67 +/- 0.05, P < 0.001) and a greater resistive index (0.61 +/- 0.04 vs. 0.53 +/- 0.04, P < 0.001) and a lower blood flow (127 +/- 42 ml/min vs. 167 +/- 37 ml/min, P < 0.03) in the middle cerebral artery compared with controls. Meal ingestion significantly increased the pulsatility index (P < 0.03) and the resistive index (P < 0.01) and decreased blood flow (P < 0.03) in the middle cerebral artery in cirrhotics but not in controls. In contrast, placebo ingestion had no effect on the hemodynamic parameters in the middle cerebral artery in the two groups., Conclusions: Results support the hypothesis that middle cerebral arterial vasoconstriction seen in cirrhotic patients is one of the cerebral artery's homeostatic responses to underfilling of the splanchnic arterial circulation.

We report a case of primary stomach carcinoma arising in the atypical epithelium, which was accompanied with ossification. The patient was a 73-year-old man, who was operated because of the possibility of advanced Type IIa carcinoma. Grossly, a large papillary polypoid lesion was recognized in the anterior wall of the antrum near the pylorus ring. Histological study revealed well differentiated tubular adenocarcinoma arising in the atypical epithelium; ossification was found in the stroma without necrosis, calcification or chondrification. Ossification arising in primary stomach carcinoma is very rare. Only five cases of ossification in primary stomach carcinoma have been reported in the world literature; two of these were Japanese patients. Probably ours is the first reported case with both stomach carcinoma and ossification arising in the atypical epithelium.

A 54-year-old male who noticed a growing mass in his right inguinal region one month earlier visited our clinic with a pigeon-egg sized, hard elastic, and movable mass with an unsmoothed surface. The removed mass was 4 X 2 X 2.5 cm, weighed 11.5 g and its cut surface was greyish white. The histopathological diagnosis was primary lipogranuloma. Postoperative course was uneventful with no sign of recurrence one year later.

The first case was a congenital renal arteriovenous fistula in a 33-year-old woman and the second case was a secondary renal arteriovenous fistula in a 67-year-old man. Both cases were seen with asymptomatic gross hematuria without any cardiovascular symptoms. In case 1, the arterial stage of right selective renal angiogram demonstrated a cirsoid type arteriovenous fistula in the right kidney; then, partial nephrectomy was performed. Angiogram of case 2 demonstrated arteriovenous fistula in the lower part of the right kidney. In this case arterial embolization was performed. After reviewing the literature, we believe that partial nephrectomy or arterial embolization is the most useful operative procedure to treat almost all cases of the cirsoid type renal A-V malformation. Conservative treatment of arteriovenous fistula has increased during the post 10 years.

Three statistical models of the individual's physiological variations, proposed by Dr Eugene K. Harris, were applied to the long-term series of individual test results in our health control system. It was found from this study that, in most cases, the homeostatic model shows the best fit to the population level from comparing correlation coefficients between the observations and estimations made based on Harris's three models. It was also found that for most cases, the homeostatic model gives the most reliable estimates in individual's level from the comparison of the chi-square values between the observations and estimations made for the individual's successive test results. Further, for a group of members with hyperglycaemia, the proportion of individuals in whom the random-walk model produced the most accurate predictions was increased to the proportion in the normal group. In the age group under 39 years of age, the autoregressive model showed a relatively high degree of predictive success, while the homeostatic model showed a relatively low degree in comparison with the results found in other age groups. From these investigations, it was found that the physiologically normal state shows strict homeostatic stability.

Tumor in the spermatic cord is infrequent. Further, among such cases, it is extremely rare to encounter a carcinoma other than a nonepithelial malignant tumor; almost all are metastatic tumors. We recently experienced an interesting case of spermatic cord tumor in a 78-year-old male. He entered our hospital with the chief complaint of a right inguinal tumor. He underwent 2 operations to excise the tumor. The extirpated tumor mass from the right spermatic cord was composed of adenocarcinoma with abundant fibrous stroma, which is considered to be metastasis from the other viscerae. We were unable to find the primary focus in spite of extensive clinical and pathological examinations. In an autopsy about four months after the 2nd operation, the tumor was confirmed to be primary intrahepatic bile duct carcinoma, which is called cholangioma. There have been only 20 cases of metastatic spermatic cord tumor reported in Japan. However, we considered that this is the 1st case caused by cholangioma in Japan and even in other countries.

Lipogranulomas developing secondarily in the genitourinary system have been reported rather frequently, but primary lipogranulomas without any past history of etiologically related conditions are rare. We report on 2 cases recently encountered in which tumors were diagnosed as sclerosing lipogranuloma on histopathologic examination.

The pathogenesis of experimental arteritis induced by Candida alkali-extract in mice was studied from the immunological, pharmacological and chemical points of view. The following results were obtained. 1) The incidence of arteritis increased in parallel with IgE antibody level of Candida, as shown by the PCA method. 2) Candida polysaccharide showed cardiotonic, coronary vasodilatatory and vasoconstrictive activities on Langendorff's and spiral preparation of rats. 3) Candida polysaccharide was shown to contain digitoxose by gas-mass spectrometry. From these results, it is suspected that this extract reacts on experimental animals by the toxic and immunological mechanisms, and that these mechanisms may be related to the occurrence of arteritis.

A case of a "so-called carcinosarcoma" of the pancreas that occurred in a 48-year-old female is reported. At autopsy, the pancreatic tumor was seen to have deeply invaded the gastric wall, causing ulceration. The pancreatic tumor consisted of two different histological components: a squamous cell carcinoma and spindle-cell sarcomatous tumor cells. Transitional areas between the two different tumor cells were also present. Since there were transitional areas and a lack of definitive electron microscopic evidence favoring a sarcoma in the sarcomatous tumor cells, the pancreatic tumor was considered to represent a "so-called carcinosarcoma".

A 19-year-old male with papillary adenocarcinoma of the seminal vesicle with cyst is reported. Cytological and histopathological studies were carried out and the relevant literature was perused. There are only 6 cases of carcinoma and 40 cases of cyst of the seminal vesicle in the Japanese literature. Moreover, ours is the first reported case of the co-existence of carcinoma and cyst in the seminal vesicle. There may be a pathogenetic relationship between the carcinoma and cyst in our case.

A boy 15 months old with diffuse infantile fibromatosis in the thigh is reported. A rapidly growing tumor in the right thigh was the only clinical manifestation, suggesting a malignant soft tissue tumor. Macroscopically, the lesion in the medial vastus muscle with an illdefined border gave a pepper and salt appearance. Histologically, the lesion was characterized by an abnormal proliferation of fibroblasts accompanied by some fibrosis, infiltration of chronic inflammatory cells, formation of lymphoid follicles, proliferation of capillaries and the presence of a small number of mature fat cells. No evidence of recurrence is found at present one and half years after operation.