BACKGROUND: Toll like receptor (TLR), an element of innate immunity, is upregulated by Ischemia/reperfusion (IR) injury and may be involved in adaptive immune response. Immunosuppressive agents may increase or attenuate IR injury and TLR expression. To explore the involvement of TLRs in hypoxic tubular injury and modification by mycophenolic acid (MPA) rapamycin (RAP), this study examined TLR expression in hypoxia-induced human renal proximal tubular epithelial cells (HK-2). METHODS: HK-2 cells were cultured in keratinocyte-SFM media supplemented with epidermal growth factor and bovine pituitary extract. The Induction of hypoxia was achieved using GasPak pouch system. TLR 2, 3, and 4 mRNA expression was analyzed by real time RT-PCR using SYBR green and TLR 4 protein expression was evaluated by Western blot analysis. MPA at concentration of 100 nM and 1uM and RAP at concentration of 20, 50, and 100 nM were added to culture medium. RESULTS: TLR4 but noTLR2 or TLR3 mRNA expressions increased in hypoxic HK-2 cells at 24 and 48 hrs. TLR4 protein expression also increased in hypoxic HK-2 cells at 24 and 48 hrs. MPA (100 nM and 1uM) and RAP (20, 50, and 100 nM) decreased hypoxia-induced TLR4 mRNA expression in HK-2 cells compared to normoxia at 24 hrs. However, TLR4 protein expression was decreased only by RAP at 20 and 50 nM. CONCLUSIONS: The results suggest that RAP may modify hypoxic renal tubular damage by decreasing TLR4-mediated inflammatory and immune reactions.
Adaptive Immunity ; Anoxia ; Blotting, Western ; Epidermal Growth Factor ; Epithelial Cells ; Humans ; Immunity, Innate ; Immunosuppressive Agents ; Mycophenolic Acid ; RNA, Messenger ; Sirolimus ; Toll-Like Receptors

This paper aims to historically re-evaluate the issues surrounding resident participation in health care and its legacy by examining the community participation in health care. In the 1970s and 1980s, community participation was one of crucial trends and a controversial topic in the international health as well as the international aid. Throughout the 1970s, local participation was regarded as one of essential elements for the access to basic healthcare and primary health care in developing countries. Community health projects which aimed to apply primary health care were implemented in rural areas and some urban areas in the 1970s and 1980s. Village Health Workers(VHWs) were a symbolic example of community participation in these projects. They consisted of local women and led health activities with simple skills in their villages. They served as a bridge between the project team and the residents. Health professionals expected them to be health leaders for “self-help” in health of their communities. In the mid-1980s, however, as the number of health facilities and professional health care workers increased, the activities of VHSs were decreased. The mixed understanding of the responsibilities and roles of VHSs among the health professionals affected the skeptical view on the achievement and effects of community participation in health care. In the mid-1980s, as the government officially organized the VHWs, the dilemma surrounding community participation intensified. When the community health projects were ended, most of the VHW organizations were also disbanded. After the projects, the spirit of community participation was only inherited by some healthcare movement organizations, such as medical cooperatives.

PURPOSE: Deposition of polymeric IgA1 in the kidney mesangium is the hallmark of IgA nephropathy, but the molecular mechanisms of IgA-mediated mesangial responses and inflammatory injuries remain poorly understood. We hypothesize that Toll-like receptor 4 (TLR4) is involved in IgA-induced mesangial cell activation. MATERIALS AND METHODS: Mouse mesangial cells were stimulated with lipopolysaccharide (LPS) (1 microg/mL), IgA (20 microg/mL), or both, and TLR4 expression was measured by real time RT-PCR and Western blot. Intracellular responses to LPS or IgA were assessed by Western blot for ERK1/2, JNK, p38 MAP kinases (MAPKs), Ikappa-Balpha degradation and fibronectin secretion. MCP-1 secretion was assessed by ELISA. Small interfering RNA (siRNA) of TLR4 was used to confirm that the effects were caused by TLR4 activity. RESULTS: LPS- or IgA-treatment upregulated the levels of TLR4 mRNA and protein in cultured MMC at 24 h. LPS and IgA induced rapid phosphorylation of MAPKs, but degradation of Ikappa-Balpha was observed only in LPS-treated MMC. LPS, but not IgA, induced increased secretion of MCP-1 and fibronectin at 24 h or 48 h. Combined LPS and IgA treatment did not cause additional increases in TLR4 mRNA and protein levels or Ikappa-Balpha degradation, and MCP-1 and fibronectin secretions were less than with LPS alone. LPS- or IgA-induced TLR4 protein levels and MAPK activation were inhibited by transfection with TLR4 siRNA. CONCLUSION: These results indicate that the activation of MAPKs and MCP-1 secretion are mediated by TLR4, at least in part, in IgA-treated mesangial cells. TLR4 is involved in mesangial cell injury by induction of pro-inflammatory cytokines in IgA nephropathy.
Animals ; Chemokine CCL2/secretion ; Enzyme-Linked Immunosorbent Assay ; Extracellular Signal-Regulated MAP Kinases/metabolism ; Fibronectins/secretion ; Glomerulonephritis, IGA/*metabolism ; I-kappa B Proteins/metabolism ; Mesangial Cells/*metabolism/secretion ; Mice ; Mice, Transgenic ; Phosphorylation ; RNA Interference ; RNA, Messenger/metabolism ; *Signal Transduction ; Toll-Like Receptor 4/antagonists & inhibitors/genetics/*metabolism

Background: The increased prevalence of obesity has led to increases in the prevalence of chronic diseases worldwide. There is interest whether probiotics have an effect on obesity, but the effectiveness and safety of only a few probiotics for the treatment of obesity have been reported. The purpose of this study was to investigate whether ingestion of Lactobacillus sakei (CJLS03) derived from kimchi causes weight loss in people with obesity. Methods: This randomized, double-blind, placebo-controlled, clinical trial involved 114 adults with a body mass index (BMI) ≥25 kg/m2 who were assigned randomly to a CJLS03 or placebo group. The groups received two allocations of either 5×109 colony-forming units of CJLS03/allocation or the equivalent vehicle for 12 weeks. Demographic and biochemical parameters, and body composition including fat and muscle mass were measured at baseline and after 12 weeks. Changes in body fat, weight, and waist circumference were compared between the two treatment groups. Adverse events were monitored during study period. Results: Body fat mass decreased by 0.2 kg in the CJLS03 group and increased by 0.6 kg in the placebo group (0.8 kg difference, P=0.018). After the 12 weeks, waist circumference was 0.8 cm smaller in the CJLS03 group than in the placebo group (P=0.013). BMI and body weight did not change after the 12 weeks. Adverse events were mild and did not differ between the two groups. Conclusion These data suggest that L. sakei (CJLS03) might help people with obesity reduce body fat mass without serious side effects (ClinicalTrials.gov: NCT03248414).

Tomato extract has been shown to exert antiplatelet activity in vitro and to change platelet function ex vivo, but with limitations. In this study, antiplatelet activity of water soluble tomato concentrate (Fruitflow I) and dry water soluble tomato concentrate (Fruitflow II) was investigated using rat platelets. Aggregation was induced by collagen and adenosine diphosphate and granule-secretion, [Ca2+]i, thromboxane B2, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) levels were examined. The activation of integrin αIIbβ3 and phosphorylation of signaling molecules, including mitogen-activated protein kinase (MAPK) and PI3K/Akt, were investigated by flow cytometry and immunoblotting, respectively. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were examined. Moreover, in vivo thrombus weight was tested by an arteriovenous shunt model. Fruitflow I and Fruitflow II significantly inhibited agonist induced platelet aggregation, adenosine triphosphate and serotonin release, [Ca2+]i, and thromboxane B2 concentration, while having no effect on cAMP and cGMP levels. Integrin αIIbβ3 activation was also significantly decreased. Moreover, both concentrates reduced phosphorylation of MAPK pathway factors such as ERK, JNK, P38, and PI3K/Akt. In vivo thrombus formation was also inhibited. Taken together, these concentrates have the potential for ethnomedicinal applications to prevent cardiovascular ailments and can be used as functional foods.
Adenosine Diphosphate ; Adenosine Monophosphate ; Adenosine Triphosphate ; Animals ; Blood Platelets* ; Cardiovascular Diseases ; Collagen ; Flow Cytometry ; Functional Food ; Guanosine Monophosphate ; Immunoblotting ; In Vitro Techniques ; Lycopersicon esculentum* ; Partial Thromboplastin Time ; Phosphorylation ; Platelet Aggregation ; Protein Kinases* ; Prothrombin Time ; Rats ; Serotonin ; Thrombosis ; Thromboxane B2 ; Water*

A double-stranded RNA (dsRNA) mycovirus was detected in malformed fruiting bodies of Pleurotus ostreatus strain ASI2792, one of bottle cultivated commercial strains of the edible oyster mushroom. The partial RNA-dependent RNA polymerase (RdRp) gene of the P. ostreatus ASI2792 mycovirus (PoV-ASI2792) was cloned, and a cDNA sequences alignment revealed that the sequence was identical to the RdRp gene of a known PoSV found in the P. ostreatus strain. To investigate the symptoms of PoV-ASI2792 infection by comparing the isogenic virus-free P. ostreatus strains with a virus-infected strain, isogenic virus-cured P. ostreatus strains were obtained by the mycelial fragmentation method for virus curing. The absence of virus was verified with gel electrophoresis after dsRNA-specific virus purification and Northern blot analysis using a partial RdRp cDNA of PoV-ASI2792. The growth rate and mycelial dry weight of virus-infected P. ostreatus strain with PoV-ASI2792 mycovirus were compared to those of three virus-free isogenic strains on 10 different media. The virus-cured strains showed distinctly higher mycelial growth rates and dry weights on all kinds of experimental culture media, with at least a 2.2-fold higher mycelial growth rate on mushroom complete media (MCM) and Hamada media, and a 2.7-fold higher mycelial dry weight on MCM and yeastmalt-glucose agar media than those of the virus-infected strain. These results suggest that the infection of PoV mycovirus has a deleterious effect on the vegetative growth of P. ostreatus.
Agar ; Agaricales* ; Blotting, Northern ; Clone Cells ; Culture Media ; DNA, Complementary ; Electrophoresis ; Fruit ; Fungal Viruses* ; Methods ; Pleurotus* ; RNA Replicase ; RNA, Double-Stranded ; Weights and Measures

BACKGROUND/OBJECTIVES: The aim of this study was to examine the validity and reproducibility of a food frequency questionnaire (FFQ) developed in Korea for breast cancer survivors. SUBJECTS/METHODS: Ninety-nine breast cancer survivors who completed an FFQ twice and three 3-day dietary records (DRs) between 2016–2017 were included. Energy and 14 nutrient intakes were calculated from FFQs and DRs. To determine the validity of the FFQ, energyadjusted de-attenuated Pearson correlations between two FFQ assessments and the average of the three 3-day DRs were calculated, and to determine reproducibility, energy-adjusted Pearson correlations and degrees of agreement were calculated between the first and second FFQ assessments. RESULTS: Correlation coefficients of validity ranged from 0.29 (protein) to 0.47 (fat) (median value = 0.36) for the FFQ assessment and from 0.20 (riboflavin) to 0.53 (calcium) (median value = 0.37) for the second. Correlation coefficients of reproducibility ranged from 0.22 (sodium) to 0.62 (carbohydrate) (median value = 0.36). Regarding FFQ reproducibilities, percentage classifications of exact agreements for energy-adjusted nutrients ranged from 27.3% (sodium) and 45.5% (fat). A median 76.8% of participants were classified into the same or adjacent quartiles, while a median of 5.6% of participants were classified in extreme quartiles. Bland–Atman plots for the majority of data points of three macronutrients, calcium and vitamins A and C fell within limits of agreement. CONCLUSIONS These results indicated that the newly developed FFQ for Korean breast cancer survivors has acceptable validity and reproducibility as compared with three 3-day DRs collected over a one-year period.

Purpose: Assessing the status of metastasis in sentinel lymph nodes (SLNs) by pathologists is an essential task for the accurate staging of breast cancer. However, histopathological evaluation of sentinel lymph nodes by a pathologist is not easy and is a tedious and time-consuming task. The purpose of this study is to review a challenge competition (HeLP 2018) to develop automated solutions for the classification of metastases in hematoxylin and eosin–stained frozen tissue sections of SLNs in breast cancer patients. Materials and Methods: A total of 297 digital slides were obtained from frozen SLN sections, which include post–neoadjuvant cases (n = 144, 48.5%) in Asan Medical Center, South Korea. The slides were divided into training, development, and validation sets. All of the imaging datasets have been manually segmented by expert pathologists. A total of 10 participants were allowed to use the Kakao challenge platform for six weeks with two P40 GPUs. The algorithms were assessed in terms of the AUC (area under receiver operating characteristic curve). Results: The top three teams showed 0.986, 0.985, and 0.945 AUCs for the development set and 0.805, 0.776, and 0.765 AUCs for the validation set. Micrometastatic tumors, neoadjuvant systemic therapy, invasive lobular carcinoma, and histologic grade 3 were associated with lower diagnostic accuracy. Conclusion In a challenge competition, accurate deep learning algorithms have been developed, which can be helpful in making frozen diagnosis of intraoperative sentinel lymph node biopsy. Whether this approach has clinical utility will require evaluation in a clinical setting