Polyphenolics from Mango (Mangifera indica L.) and Pomegranate (Punica granatum L.) Suppress Inflammation in in vivo and in vitro Models for Colitis

Ulcerative colitis is a chronic inflammation of the large intestine, and it may increase risk of human colorectal cancer. Polyphenolics from mango and pomegranate have been shown to have potent anti-inflammatory properties, thus they could be the potential therapeutic agents for colitis. However, the mechanism underlying these effects of polyphenolics has not yet been elucidated. To determine the anti-inflammatory effects and possible mechanisms of polyphenolics from mango (gallic acid and gallotannins), and pomegranate (ellagic acid and ellagitannins) in dextran sodium sulfate (DSS)-induced colitis in rats, Sprague Dawley rats were administered control, mango, or pomegranate juice, and were exposed to three cycles of 3% DSS followed by 2-week recovery period. Colon inflammation and injury scores were assessed, and cell proliferation was evaluated by immunohistochemical detection of Ki-67. The mRNA and protein expressions involved in the inflammatory response and the mTOR pathway were analyzed by qRT-PCR, low density arrays, western blot analysis and multiplex bead assay. The involvement of miRNAs was additionally investigated with the antagomiR-126 and antagomiR-145 in lipopolysaccharide (LPS)-treated CCD-18Co, non-cancer colon fibroblasts cell lines. Both mango and pomegranate showed anti-inflammatory properties in vitro and in vivo. Mango and pomegranate juice reduced DSS-induced colon inflammation score (41% and 50%) and cell proliferative index (38% and 36%) during chronic colitis in rats compared to control juice. Mango and pomegranate juice significantly attenuated the pro-inflammatory cytokines, CRP, TNF-alpha, IL-1beta, GM-CSF, and IL-6 levels in colonic tissues. In addition, mango and pomegranate juice suppressed COX-2 and iNOS mRNA and protein expressions. Mango juice suppressed HIF-1alpha by decreasing the PI3K(p85beta)/AKT-mTOR signaling axis via up-regulation of miR-126, while pomegranate decreased p70S6K and HIF-1alpha by up-regulating miR-145. The interactions of mango with miR-126/PI3K(p85beta) and pomegranate with miR-145/p70S6K1 were additionally identified in CCD-18Co cells, where mango and pomegranate extract reversed the effect of the antagomiR. In addition, the modulation of microbiota composition (Blautia, Fusobacterium, and Ruminococcaceae) by pomegranate and short-chain fatty acids (SCFA; Isovalerate and valerate) production by mango may be involved in at least in part the anti-inflammatory effects of polyphenolics. These results suggest that both mango and pomegranate polyphenolics seem to have potential in the prevention and mitigation of colon inflammation

Optimisation and Application of Comparative Genomic Hybridisation (CGH) in Cancer Cytogenetics

The aim of this study was to assess to the applicability of the CGH technique to analysis of various kinds of malignancies in particular hematological malignancies and solid tumours, and to optimise a CGH technique for use on very small amounts of DNA from paraffin embedded tissue. During the initial part of the study several FISH experiments were also performed. First, interphase FISH with selected chromosome specific alpha satellite probes was applied to 22 cases with acute lymphoblastic leukemia, to investigate whether this could contribute to the detection of hidden hyperdiploidy or detect aneuploidy in cases where cytogenetic analysis had failed. Then CGH and interphase FISH were both performed in 19 cases with acute lymphoblastic leukemia to compare the efficiency of the two approaches. CGH showed potential as a comprehensive screening method for detection of hyperdiploid cases and additionally unbalanced DNA copy number changes including one amplification on 6p23-pter were detected. Interphase FISH also might be a complementary method to confirm CGH results, especially in hyperdiploid or hypotriploid cases which failed by conventional karyotyping. CGH analysis in twenty-two cases of AML, MDS or MDS in transformation, revealed that net gains and losses of chromosomal material could be detected more simply and accurately by CGH than by conventional karyotyping in cases with a complex karyotype. CGH followed by FISH using specific probes which were chosen according to discordance between CGH and conventional karyotyping could identify the origin of marker chromosomes, and CGH also could detect amplification sites where candidate genes related to pathogenesis of cancer might be harbored. A further fourteen cases of other hematological malignancies were analysed by CGH and another hidden abnormality which was missed by conventional karyotyping was detected in 1 case. Next, CGH experiments moved to fresh frozen solid tumours. Five cases of well differentiated liposarcoma and seventeen cases of malignant melanoma were analysed. In well differentiated liposarcoma CGH could detect additional amplifications including one amplification (3ql2-ql3.3) which was hitherto unreported and helped to evaluate the composition of marker chromosomes. In cases with malignant melanoma, CGH could identify frequently involved regions of gains and losses, the inter-relationship between abnormalities, and found three new sites of amplification (1 pl 1.1-p12, 3p24, and 22) and amplification on 17q24-qter in malignant melanoma. The CGH technique was next applied to very small amounts of DNA from paraffin-embedded tissue of solid tumours. For success of this experiment, several steps from the extraction of DNA to CGH itself had to be optimised. Although it would have been ideal to move to the next step after completing the optimisation of one step, most experiments involving the extraction of DNA went on to attempt CGH because these test samples were precious material and the final goal of this experiment was to obtain successful CGH result. For obtaining an adequate quantity of high quality of DNA, fifteen different parameters were investigated to discover the optimal method for extraction of DNA from paraffin-embedded tissue. According to this optimisation process, in a very small sample (1 x 7mum) a single step cell lysis method (without phenol/chloroform extraction) and higher dosage of proteinase K (1-2 mg/ml) with shorter duration (≤24 hrs) showed better results. For whole genome amplification, two methods - primer extension preamplification (PEP) and degenerate oligonucleotide primed-PCR (DOP-PCR) were used. DOP-PCR was performed with 8 different minor modifications. The DOP-PCR method without disruption from start to finish was the better method for avoidance of contamination and cases using thermosequenase showed better results than those using combination of topoisomerase and T7 sequenase. PEP was performed with several modifications to assess if whole genome amplification (WGA) using PEP could produce analysable CGH images and if so what volume of starting template DNA could produce CGH images by this method. Another aim was to evaluate the difference of results in comparison with DOP-PCR. Although PEP produced some successful CGH images, it showed some limitation to amplify a sufficient amount of DNA to produce reliable CGH from very small samples. By comparing results between both methods, there are some discrepancies of amplification pattern. After the optimisation process was almost completed, this technique was applied to three kinds of serially diluted samples (DNA from normal female blood, abnormal bone marrow with AML/MDS, and thick-sectioned paraffin-embedded ovarian cancer tissue). These experiments showed that the smallest amount of template DNA which could produce reliable CGH images was above 10 pg of DNA from normal female blood, above 50 pg of DNA from abnormal bone marrow with complex chromosome aberrations, and above 100 pg of DNA from thick sectioned paraffin-embedded ovarian cancer tissue. This study also revealed that the application of the same amplification and labelling method to both test and control DNA was the ideal method for decreasing bias developed during the amplification and labelling process

Intention to Use Long-Term Care Facilities: Differences between Korean Pre-elderly and Korean Baby-boomers

With the rapidly increasing number of older adults, dealing with long-term care (LTC) needs becomes an emerging issue in South Korea. This study aims to examine factors affecting the intention to use longtermcare facilities with two groups of young-old adults: (1) Korean pre-elderly (KPE) and (2) Korean babyboomers (KBB). Guided by Andersen's behavioral model of health service use and prior research, predisposing characters, enabling resources, need factors, availabilities of informal care and self-care activities were used as predictors. In the final analyses, 803 KPE and 966 KBB were included. The results of logistic regression analyses showed different findings in two groups. Age, education, spouse's physicalhealth, and self-care activities for relationship with family and friends are significantly associated with intention to use LTC facilities among KPE. However, income, physical health of respondents, and relationship satisfaction with children are significantly related to intention of use LTC facilities in the group of KBB. This study suggests different LTC needs between KPE and KBB. Health care professionals and policy makers need to consider such differences to provide quality LTC care for them

The Extraction of Knowledge Factors of Teachers for Physical Computing Education

In informatics, physical computing focuses on interactions to realize the real world as a computing system. From 2018, how to teach the physical computing in informatics as a mandatory subject is important. The purpose of this study is to analyze the problems in the physical computing education recognized by secondary school informatics teachers and to provide implications for effective programming education. First, we extracted related keywords of physical computing in the 2015 revised informatics curriculum and science curriculum. Second, extracted keywords are classified into hardware and programming. Third, we developed a questionnaire item suitable for classification keywords. Finally, web surveys were conducted and analyzed for in-service and pre-service secondary school informatics teachers. As a result of the research, it was confirmed that the informatics teachers recognized that physical computing education was helpful for programming education. However, a large proportion of the member's lack of training time and receive appropriate education and training programs, hardware, reduced the level of knowledge about the physical computing element content

Two-point discrimination values vary depending on test site, sex and test modality in the orofacial region: a preliminary study

The two-point discrimination (TPD) test is one of the most commonly used neurosensory tests to assess mechanoperception in the clinical settings. While there have been numerous studies of functional sensibility of the hand using TPD test, there have been relatively not enough reports on TPD in the orofacial region. Objective The aims of the present study were to determine the normal values of TPD in the six trigeminal sites (the forehead, cheek, mentum, upper lip, lower lip, and the tongue tip) and to investigate the effect of the site, sex, and test modality on the TPD perception. Material and Methods Forty healthy volunteers consisting of age-matched men (20) and women (20) with a mean age of 27.1 years were recruited. One examiner performed the TPD test using a simple hand-operated device, i.e., by drawing compass with a blunt or sharp-pointed tip. The static TPD with a blunt-pointed tip (STPDB), moving TPD with a blunt-pointed tip (MTPDB), and static TPD with a sharp-pointed tip (STPDS) were measured. The predictors were the site, sex, and test modality, and the outcome variable was the TPD value. Three-way ANOVA was used for statistics. Results The analysis showed a significant effect of the site, sex and test modality on the TPD values. Significant differences between the test sites were observed with the descending order from the forehead and cheek>;mentum>;upper lip and lower lip>;tongue tip and index finger. Women showed lower TPD values than those of men. The STPDS measurements were consistently lower than those of the STPDB and MTPDB. Conclusions The normal values of TPD in this study suggest that the cheek and forehead were less sensitive than other regions evaluated and women were more sensitive than men. The STPDS was the most sensitive test modality

IRT5 Probiotics Changes Immune Modulatory Protein Expression in the Extraorbital Lacrimal Glands of an Autoimmune Dry Eye Mouse Model

PURPOSE. While the association between the gut microbiome and the immune system has been studied in autoimmune disorders, little is known about ocular disease. Previously we reported that IRT5, a mixture of five probiotic strains, could suppress autoimmune dry eye. In this study, we investigated the mechanism by which IRT5 performs its immunomodulatory function in a mouse model of autoimmune dry eye. METHODS. NOD.B10.H2b mice were used as an autoimmune dry eye model. Either IRT5 or PBS was gavaged orally for 3 weeks, with or without 5 days of antibiotic pretreatment. The effects on clinical features, extraorbital lacrimal gland and spleen proteins, and fecal microbiota were analyzed. RESULTS. The ocular staining score was lower, and tear secretion was higher, in the IRT5-treated groups than in the PBS-treated groups. After IRT5 treatment, the downregulated lacrimal gland proteins were enriched in the biological processes of defense response and immune system process. The relative abundances of 33 operational taxonomic units were higher, and 53 were lower, in the feces of the IRT5-treated groups than in those of the PBS-treated groups. IRT5 administration without antibiotic pretreatment also showed immunomodulatory functions with increases in the Lactobacillus helveticus group and Lactobacillus hamsteri. Additional proteomic assays revealed a decrease of proteins related to antigen-presenting processes in the CD11b(+) and CD11c(+) cells of spleen in the IRT5-treated groups. CONCLUSIONS. Changes in the gut microbiome after IRT5 treatment improved clinical manifestations in the autoimmune dry eye model via the downregulation of antigen-presenting processes in immune networks.11Ysciescopu

Death Attitudes Among Middle-Aged Koreans: Role of End-of-Life Care Planning and Death Experiences

The purpose of this study was to examine factors affecting death attitudes among middle-aged Koreans. In addition, the study explored the interaction effect between knowledge about end-of-life care planning and the experience of death of family or friends on death attitudes. The sample was obtained from a national survey with middle-aged adults in South Korea (n = 2,026). Multivariate regression analysis revealed significant main effects and an interaction effect between knowledge about end-of-life care planning and the experience of death on death attitudes. Greater knowledge of end-of-life care planning was associated with more positive attitudes toward death; however, the effect was stronger for those who had not experienced the death of family or friends. Being older and having greater life satisfaction were also associated with more positive attitudes toward death. This study suggests that end-of-life education can help middle-aged adults embrace the final stage of life and prepare for their own death

Keeping Customers Shopping in the Store: Interrelationships among Store Attributes, Shopping Enjoyment, Place Attachment, and Store Loyalty

With consumers shifting to Internet and mobile shopping, the demand for brick and mortar store shopping has decreased and in response, several major U.S. retail chains have downsized the number of their physical retail operations (Farfan, 2013). Yet, retailers have not completely said good bye to stores. Indeed there may be several key store attributes that retailers’ can focus on and continue to develop if they are interested in keeping customers shopping and buying in brick and mortar stores

Extensive Emphysematous Pyelonephritis in a Nondiabetic Female Cat - Treatment with Unilateral Nephroureterectomy

Background: Emphysematous pyelonephritis (EPN) is an acute, severe necrotizing infection of the renal parenchyma and surrounding tissues that results in gas formation in the kidney, collecting system, or surroundings. EPN is a rare condition in veterinary medicine and occurs most frequently in dogs with diabetes mellitus. Although the prognosis of medical management in animals is poor, the standardized treatment protocol according to EPN severity is unclear. This report describes the first case of a nondiabetic female cat with extensive EPN and good prognosis following direct nephroureterectomy (NU). Case: A 10-year-old spayed female cat presented with the chief complaint of an acute loss of weight within 1 week, vomiting, and disorientation including stumbling, discoordination, circling, wobbling, head tilting, and difficulties in standing. At presentation, the patient had a body condition score of 1/9 and weighed 2.6 kg. Blood examination revealed leukocytosis, anemia, and hypoproteinemia. Abdominal radiography revealed severely decreased serosal details. A massive gas silhouette observed in the peritoneal and retroperitoneal cavities, was diagnosed as abdominal free gas. Abdominal ultrasound showed an accumulation of moderately anechoic fluid mixed with gas and cyst-like capsules around the left kidney. Left partial ureteral obstruction and dilation were also observed. Computed tomography (CT) was performed without sedatives or anesthetic drugs. The findings showed severe inflammatory changes in the peritoneum and a loss of the normal inner structure in the left kidney. A pyelogram of the left kidney was not observed after injection of the contrast material. Diffuse fat stranding and free gas observed in the mesentery of the entire abdominal cavity and around the left kidney were considered septic peritonitis. Urinalysis revealed proteinuria and hematuria. Numerous neutrophils with rod-type bacteria were observed in the ascites. Following diagnostic examinations, the patient was diagnosed with extensive left EPN, including inflammatory ascites and abdominal free gas. Therefore, emergency NU of the nonfunctional left kidney and ruptured ureter and thorough abdominal lavage were conducted. Diffuse inflammation and a nephrolith were observed in the section of the harvested kidney. The nephrolith was composed of 100% calcium oxalate monohydrate. The real-time polymerase chain reaction (RT-PCR) test for feline infectious peritonitis (FIP) was negative. Escherichia coli was detected in the ascites, and antibiotic therapy was administered following the antibiotic sensitivity test. The histological findings from the left kidney and ureter included marked chronic inflammation and fibrosis. The patient was discharged 4 days after surgery. During the 8-month follow-up period, the patient’s condition improved. Discussion: This was a unique case of EPN in a nondiabetic cat and the first reported case of EPN with a ruptured ureter, including abdominal free gas, inflammatory ascites, and peritonitis. This patient had a bacterial urinary tract infection with E. coli, which is the most frequently isolated pathogen in humans. This gas-forming bacteria produced a massive amount of gas and inflammation that were considered to have ruptured the urinary tract, so that the gas was released into the abdomen. This case corresponded to class 3B, with two risk factors according to the human EPN classification system. Direct NU and abdominal lavage were performed as emergency surgeries. The patient stabilized gradually and showed a good prognosis. Immediate surgical intervention is recommended in animal patients showing the extensive EPN stage. Keywords: kidney, nephroureterectomy, emphysematous pyelonephritis, peritonitis, cat, E. coli.