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Although many aquaporin (AQP) transcripts have been demonstrated to express in the female reproductive tract, the defined localizations and functions of AQP subtype proteins remain unclear. In this study, we investigated the expression of AQP1, AQP3, AQP5, AQP6, and AQP9 proteins in female reproductive tract of mouse and characterized their precise localizations at the cellular and subcellular levels. Immunofluorescence analyses for AQP1, AQP3, AQP6, and AQP9 showed that these proteins were abundantly expressed in female reproductive tract and that intense immunoreactivities were observed in mucosa epithelial cells with a subtype-specific pattern. The most abundant aquaporin in both vagina and uterine cervix was AQP3. Each of AQP1, AQP3, AQP6, and AQP9 exhibited its distinct distribution in stratified squamous or columnar epithelial cells. AQP9 expression was predominant in oviduct and ovary. AQP1, AQP3, AQP6, and AQP9 proteins were mostly seen in apical membrane of ciliated epithelial cells of the oviduct as well as in both granulosa and theca cells of ovarian follicles. Most of AQP subtypes were also expressed in surface epithelial cells and glandular cells of endometrium in the uterus, but their expression levels were relatively lower than those observed in the vagina, uterine cervix, oviduct and ovary. This is the first study to investigate the expression and localization of 5 AQP subtype proteins simultaneously in female reproductive tract of mouse. Our results suggest that AQP subtypes work together to transport water and glycerol efficiently across the mucosa epithelia for lubrication, proliferation, energy metabolism and pH regulation in female reproductive tract., (© Copyright 2020 The Korean Society of Developmental Biology.)

Histidine triad nucleotide-binding protein 1 (HINT1), which belongs to the evolutionarily conserved HIT superfamily, has been shown to possess a tumor-suppressive function by binding to and inhibiting several oncogenic transcription factors, such as β-catenin and microphthalmia transcription factor (MITF), in various types of cancer cells. However, the regulatory mechanism that mediates the binding capacity of HINT1 for partner transcription factors remains elusive. Here, we report that HINT1 is acetylated by CBP at K21 and K30 and deacetylated by SIRT1. Deacetylation of HINT1 by SIRT1 increases the capacity of HINT1 to bind to β-catenin or MITF. As a result, the tumor-suppressive function of HINT1 is increased. In support of this, the deacetylation mimetic HINT1 mutant HINT1 2KR was found to significantly reduce cellular proliferation in colon cancer and melanoma cells and tumorigenesis in xenograft assays. Thus, this study reveals an acetylation-dependent regulatory mechanism that governs the tumor-suppressive function of HINT1.

Dendritic cells (DCs) are potent antigen-presenting cells that link the innate and adaptive immune responses; as such they play pivotal roles in initiation and progression of rheumatoid arthritis (RA). Here, we report that the tonicity-responsive enhancer-binding protein (TonEBP or NFAT5), a Rel family protein involved in the pathogenesis of autoimmune disease and inflammation, is required for maturation and function of DCs. Myeloid cell-specific TonEBP deletion reduces disease severity in a murine model of collagen-induced arthritis; it also inhibits maturation of DCs and differentiation of pathogenic Th1 and Th17 cells in vivo. Upon stimulation by TLR4, TonEBP promotes surface expression of major histocompatibility complex class II and co-stimulatory molecules via p38 mitogen-activated protein kinase. This is followed by DC-mediated differentiation of pro-inflammatory Th1 and Th17 cells. Taken together, these findings provide mechanistic basis for the pathogenic role of TonEBP in RA and possibly other autoimmune diseases.

Alverine, a smooth muscle relaxant, is used to relieve cramps or spasms of the stomach and intestine. Although the effects of alverine on spontaneous and induced contractile activity are well known, its anti-inflammatory activity has not been fully evaluated. In this study, we investigated the anti-inflammatory effects of alverine in vitro and in vivo. The production of nitric oxide (NO) in RAW264.7 cells activated by lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid (poly (I:C)) was reduced by alverine. The mRNA expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor-α (TNF-α) was also dose-dependently inhibited by treatment with alverine. In reporter gene assays, alverine clearly decreased luciferase activity, mediated by the transcription factor nuclear factor κB (NF-κB) in TIR-domain-containing adapter-inducing interferon-β (TRIF)- or MyD88-overexpressing HEK293 cells. Additionally, phosphorylation of NF-κB subunits and upstream signaling molecules, including p65, p50, AKT, IκBα, and Src was downregulated by 200 μM of alverine in LPS-treated RAW264.7 cells. Using immunoblotting and cellular thermal shift assays (CETSAs), Src was identified as the target of alverine in its anti-inflammatory response. In addition, HCl/EtOH-stimulated gastric ulcers in mice were ameliorated by alverine at doses of 100 and 200 mg/kg. In conclusion, alverine reduced inflammatory responses by targeting Src in the NF-κB pathway, and these findings provide new insights into the development of anti-inflammatory drugs.

Isolated complex III (CIII) deficiencies are among the least frequently diagnosed mitochondrial disorders. Clinical symptoms range from isolated myopathy to severe multi-systemic disorders with early death and disability. To date, we know of pathogenic variants in genes encoding five out of 10 subunits and five out of 13 assembly factors of CIII. Here we describe rare bi-allelic variants in the gene of a catalytic subunit of CIII, UQCRFS1, which encodes the Rieske iron-sulfur protein, in two unrelated individuals. Affected children presented with low CIII activity in fibroblasts, lactic acidosis, fetal bradycardia, hypertrophic cardiomyopathy, and alopecia totalis. Studies in proband-derived fibroblasts showed a deleterious effect of the variants on UQCRFS1 protein abundance, mitochondrial import, CIII assembly, and cellular respiration. Complementation studies via lentiviral transduction and overexpression of wild-type UQCRFS1 restored mitochondrial function and rescued the cellular phenotype, confirming UQCRFS1 variants as causative for CIII deficiency. We demonstrate that mutations in UQCRFS1 can cause mitochondrial disease, and our results thereby expand the clinical and mutational spectrum of CIII deficiencies., (Copyright © 2019 American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.)

The fermentation of Korean red ginseng (RG) increases the bioavailability and efficacy of RG, which has a protective role in various diseases. However, the ginsenoside-specific molecular mechanism of the fermented RG with Cordyceps militaris (CRG) has not been elucidated in non-alcoholic fatty liver disease (NAFLD). A mouse model of NAFLD was induced by a fast-food diet (FFD) and treated with CRG (100 or 300 mg/kg) for the last 8 weeks. CRG-mediated signaling was assessed in the liver cells isolated from mice. CRG administration significantly reduced the FFD-induced steatosis, liver injury, and inflammation, indicating that CRG confers protective effects against NAFLD. Of note, an extract of CRG contains a significantly increased amount of ginsenosides (Rd and Rg3) after bioconversion compared with that of conventional RG. Moreover, in vitro treatment with Rd or Rg3 produced anti-steatotic effects in primary hepatocytes. Mechanistically, CRG protected palmitate-induced activation of mTORC1 and subsequent inhibition of mitophagy and PPARα signaling. Similar to that noted in hepatocytes, CRG exerted anti-inflammatory activity through mTORC1 inhibition-mediated M2 polarization. In conclusion, CRG inhibits lipid-mediated pathologic activation of mTORC1 in hepatocytes and macrophages, which in turn prevents NAFLD development. Thus, the administration of CRG may be an alternative for the prevention of NAFLD., Competing Interests: The authors declare that they have no conflict of interest.

The bioelectrochemical anaerobic nitrogen removal was demonstrated in an anaerobic batch reactor equipped with a pair of polarized bioelectrodes. The bioelectrochemical reactor was operated in sequential batch mode after inoculating activated sludge and polarizing the electrode to 0.6 V. The medium contains ammonium, nitrite, alkalinity and trace minerals, but no organic carbon source. By the repetitive sequential operation, simultaneous removals of ammonium, nitrite and alkalinity were improved, and the electrochemical activity of the bulk sludge was confirmed from the redox peaks of the cyclic voltammogram. This indicates that ammonia oxidizing exoelectrogens (AOE) and denitritating electrotrophs (DNE) were enriched more in the bulk solution. Biogas production that mainly consisted of nitrogen was observed from the bioelectrochemical reactor, and the minor components in the biogas were methane and carbon dioxide. This demonstrates that AOE use nitrite as an electron acceptor to oxidize ammonia. The requirements of nitrite and alkalinity for the removal of ammonia nitrogen are around 0.72 mg NO 2 -N/mg NH 4 -N and 1.73 mg as CaCO 3 /mg NH 4 -N, respectively, and nitrate was not produced as a by-product. The bacterial groups involved in the bioelectrochemical nitrogen removal are electroactive autotrophs and can be enriched from activated sludge by polarized electrode. This bioelectrochemical ammonia oxidation is a novel approach recommended for treatment of nitrogen-rich wastewater., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: Photodynamic diagnosis (PDD) is a preoperative procedure that simplifies Mohs micrographic surgery (MMS) by allowing more accurate demarcation of neoplastic lesions involving the skin. After topical application of aminolevulinic acid (ALA), protoporphyrin IX (PpIX) is selectively induced in neoplastic cells with a high tumor-to-surrounding tissue ratio and can be visualized after excitation using a Wood's lamp., Objectives: To evaluate the efficacy of PpIX fluorescence images in accurately determining the boundaries of tumors during MMS., Methods: A retrospective evaluation of biopsy-proven basal cell carcinoma (BCC; n=163), squamous cell carcinoma (SCC; n=51), and Bowen's disease (n=8) in 222 patients treated with MMS was conducted. In the PDD group, 20% ALA ointment was applied 6 hours before the surgery, and then the boundaries were mapped based on the red fluorescence of the Wood's lamp in the operating room. Each of the PDD (n=142) and non-PDD (n=80) groups was surgically staged and compared., Results: The average number of stages was 1.59, with more in the PDD group (1.66) than the non-PDD group (1.44) (p=.05). The average number of stages was 1.77 in the PDD group and 1.50 in the non-PDD group (p=.06) for BCC, was 1.40 in the PDD group and 1.37 in the non-PDD group (p=.90) for SCC, and 1.25 in the PDD group and 1.00 in the non-PDD group (p=.36) for Bowen's disease., Conclusions: In previous studies, the fluorescence of induced porphyrins was effective in detecting and delineating neoplastic skin areas, but, in the present study, PDD did not support surgical efficacy during MMS., (© 2010 by the American Society for Dermatologic Surgery, Inc.)

Aims: Although heme oxygenase-1 (HO-1) is involved in osteoblastic differentiation, the HO-1- and odontoblastic differentiation-inducing effects of mechanical stress (MS) have not been clarified in human dental pulp cells (HDPCs). In this study, we examined the effects of MS on the odontoblastic differentiation of immortalized HDPCs and on the primary intracellular signaling pathways, including the HO-1 pathway, implicated in this differentiation., Main Methods: A Flexercell strain unit was used to generate cyclic tensile strain in HDPCs. Expressions of mRNAs encoding HO-1 and HDPC differentiation markers, such as osteopontin (OPN), bone sialoprotein (BSP), dentin sialophosphoprotein (DSPP), and dentin matrix-protein-1 (DMP-1), were evaluated using the reverse transcription-polymerase chain reaction. Expression of the NF-E2-related transcription factor 2 (Nrf2) protein was analyzed by Western blotting., Key Findings: MS significantly increased the expression of HO-1, OPN, BSP, DSPP, and DMP-1 mRNAs in HDPCs. HO-1 silencing and inhibitors of HO-1, p38 MAPK, ERK, phosphoinositide 3-kinase, and nuclear factor-kappaB (NF-kappaB) all attenuated MS-stimulated differentiation. The MS-induced nuclear translocation of Nrf2 was suppressed by inhibitors of PI3K and NF-kappaB., Significance: Collectively, these results provide the first evidence that MS stimulates odontoblastic differentiation of HDPCs via modulation of the Nrf2-mediated HO-1 pathway., (Copyright 2009 Elsevier Inc. All rights reserved.)