Nitrogen dioxide increases the risk of disease progression in idiopathic pulmonary fibrosis.

BACKGROUND AND OBJECTIVE: Air pollution affects clinical course and prognosis of idiopathic pulmonary fibrosis (IPF). However, the effect of individual exposure to air pollutants on disease progression is unclear. We aimed to identify the effect of individual exposure to nitrogen dioxide (NO2 ) and particulate matter (aerodynamic diameter ≤ 10 µm [PM10 ]) on disease progression in patients with IPF. METHODS: The serial lung function data of 946 IPF patients (mean age: 65.4 years, male: 80.9%) were analysed. Individual-level long-term exposures to NO2 and PM10 at the residential addresses of patients were estimated using a national-scale exposure prediction model, constructed based on air quality regulatory monitoring data. Progression was defined as a relative decline (≥10%) in forced vital capacity. Individual- and area-level covariates were adjusted in the primary analysis model. RESULTS: Overall, 547 patients (57.8%) experienced progression during a median follow-up of 1.0 year (interquartile range: 0.4-2.6 years). In the primary model, a 10-ppb increase in NO2 concentration was associated with a 10.5% increase in the risk of progression (hazard ratio [HR] = 1.105; 95% CI = 1.000-1.219) in patients with IPF. There was also an increasing trend of progression in patients with IPF according to the second to fourth quartiles of NO2 (Q2 [HR = 1.299; 95% CI = 0.972-1.735], Q3 [1.409; 1.001-1.984], Q4 [1.598; 1.106-2.310]) compared to the first quartile. We found no association between PM10 and progression in IPF patients. CONCLUSION: Our data suggest that increased individual exposure to NO2 can increase the risk of progression in patients with IPF.

Different Mortality Risks of Long-Term Exposure to Particulate Matter across Different Cancer Sites.

Particulate matter (PM) air pollution has challenged the global community and the International Agency for Research on Cancer (IARC) classified airborne particulate matter as carcinogenic to humans. However, while most studies of cancer examined a single cancer type using different cohorts, few studies compared the associations of PM between different cancer types. We aimed to compare the association of long-term exposure to PM (PM10 and PM2.5) and cancer mortality across 17 different types of cancer using a population-based cohort in the Seoul Metropolitan Area (SMA), South Korea; Our study population includes 87,608 subjects (mean age: 46.58 years) residing in the SMA from the National Health Insurance Services-National Sample cohort (NHIS-NSC) and followed up for 2007-2015. We used the time-dependent Cox proportional hazards model to estimate hazard ratios (HRs) and 95% confidence intervals (95% CIs) of each cancer mortality per 10 µg/m3 increase in PM concentrations, after adjusting for individual and areal characteristics. During eight years of follow-up, 1487 people died with any of 17 cancer types. Lung cancer death was the highest, followed by liver and stomach cancer. Although we did not find the association for all cancer types, possibly because of limited cancer cases, HRs of PM2.5 were relatively high for lung, stomach, pancreas, non-Hodgkin's lymphoma, prostate, esophagus, oral and pharynx, and brain cancer mortality (HRs = 1.44-7.14). High HRs for pancreas, non-Hodgkin's lymphoma, esophagus, and oral and pharynx cancer were also seen for PM10; our findings suggest PM air pollution as a potential risk factor of cancer mortality for upper digestive tracts, mouth, pancreas, and non-Hodgkin's lymphoma in a highly urbanized population with high exposure to PM for a long time.

Association between long-term exposure to high levels of ambient air pollution and incidence of lung cancer in a population-based cohort.

Although outdoor air pollution including particulate matter (PM) was classified as carcinogenic to humans based on accumulating epidemiological evidence, these findings were suggested mostly from low-dose environments in North America and Europe. We aimed to examine the association of long-term exposure to PM ≤ 10 and 2.5 µm in diameter (PM10 and PM2.5) and nitrogen dioxide (NO2) with lung cancer incidence using a population-based cohort in the Seoul Metropolitan Area (SMA), South Korea. Our study included 83,478 people residing in the SMA and followed up for 2007-2015 from the National Health Insurance Service-National Sample Cohort. This cohort was constructed based on the National Health Insurance database that contains sociodemographic and medical information under universal health coverage. Individual long-term concentrations of PM10, PM2.5, and NO2 were estimated at people's district-level and annually-updated residential addresses for the previous 5 years by using previously-validated prediction models. We applied a time-dependent Cox proportional hazards model and estimated hazard ratios (HRs) per 10 µg/m3 and 10 ppb increases in PM and NO2, respectively, after adjusting for individual characteristics. During 9 years of follow-up, 489 lung cancer new cases occurred (714,012 person-year). The adjusted HRs for PM10 were greater than 1 but statistically non-significant (HR = 1.15; 95% CI = 0.88-1.52). We also did not find associations for PM2.5 and NO2. Despite null associations for the total population, our subgroup analysis suggested associations with PM in family members with cancer (PM10: HR = 2.59, 95% CI = 1.26-5.32; PM2.5: 5.55, 1.09-27.91) and in those who have smoked more than 1 pack per day (1.77, 0.96-3.25; 3.81, 1.00-14.44) or for less than 20 years (2.81; 1.13-7.07; 2.02, 0.21-19.23). Our study based on a highly urbanized population exposed to relatively high air pollution provides no evidence of the association between PM and lung cancer incidence in the total population but indicates the potential susceptibility in heavy smokers for relatively short periods and family members of cancer patients. Future studies should re-examine the association using improved exposure assessment and extended population.

Nitrogen dioxide increases the risk of mortality in idiopathic pulmonary fibrosis.

Ambient air pollution is associated with the prognosis of idiopathic pulmonary fibrosis (IPF) patients. We aimed to identify the impacts of individual exposure to particulate matter with a 50% cut-off aerodynamic diameter of 10 µm (PM10) and nitrogen dioxide (NO2) on IPF patients' mortality.1114 patients (mean age 65.7 years; male 80.5%) diagnosed with IPF between 1995 and 2016 were included in this study. Individual-level long-term concentrations of PM10 and NO2 at residential addresses of patients were estimated using a national-scale exposure prediction model. The effect of PM10 and NO2 on mortality was estimated using a Cox proportional hazards model adjusted for individual- and area-level covariates.The median follow-up period was 3.8 years and 69.5% of the patients died or underwent lung transplantation. When adjusted for individual- and area-level covariates, a 10 ppb increase in NO2 concentration was associated with a 17% increase in mortality (hazard ratio (HR) 1.172, 95% CI 1.030-1.344; p=0.016). When IPF patients were stratified by age (≥65 versus <65 years) or by sex, NO2 was a significant prognostic factor for mortality in the elderly (HR 1.331, 95% CI 1.010-1.598; p=0.010). When stratified by age and sex jointly, NO2 showed the stronger association with mortality in elderly males (HR 1.305, 95% CI 1.072-1.598; p=0.008) than in other groups. PM10 was not associated with IPF mortality in all patients and in subgroups stratified by age or sex.Our findings suggest that increased exposure to NO2 can increase the risk of mortality in patients with IPF, specifically in elderly males.

In vitro and in vivo inhibition of Helicobacter pylori by Lactobacilllus paracasei HP7.

The efficacy of standard therapeutic strategies for Helicobacter pylori (H. pylori) infection is decreasing over time due to the emergence of drug-resistant strains. As an alternative, the present study investigated the capacity of Lactobacilllus paracasei (L. paracasei) HP7, isolated from kimchi, to inhibit H. pylori growth. The effects of L. paracasei HP7 on H. pylori adhesion and H. pylori-induced inflammation were examined in AGS human gastric adenocarcinoma epithelial cells and a mouse model of H. pylori SS1 infection. L. paracasei HP7 reduced H. pylori adhesion to AGS cells and suppressed the inflammatory response in infected cells by downregulating interleukin-8. H. pylori colonization in the stomach of C57BL/6 mice was demonstrated by rapid urease test, and results showed significant decrease in mice post-treated with L. paracasei HP7. Additionally, L. paracasei HP7 decreased gastric inflammation and epithelial lesions in the stomach of H. pylori-infected mice. These results demonstrate that L. paracasei HP7 treatment can inhibit H. pylori growth and is thus a promising treatment for patients with gastric symptoms such as gastritis that are caused by H. pylori infection.

Multiple roles of RARRES1 in prostate cancer: Autophagy induction and angiogenesis inhibition.

BACKGROUND: Prostate cancer (PCa) poses a major health concern in men worldwide. Retinoic Acid Receptor Responder (RARRES1)/ Tazarotene-induced gene-1 (TIG-1) is a putative tumor suppressor gene that exerts its tumor suppressor function via unknown mechanisms. Epigenetic silencing of RARRES1 leads to its loss in several types of cancer, including PCa. Determining the molecular mechanisms that mediate the tumor suppressor role of RARRES1 in PCa is the focus of our study. FINDINGS: Our data indicates that RARRES1 over expression in PCa cell lines represses mitogen-activated protein kinase (MAPK) activation. RARRES1 expression induces the levels of autophagy-related genes, beclin, ATG3 and increases LC3B-II conversion. A significant induction of SIRT1 along with mTOR inhibition is noted on RARRES1 expression. Furthermore, RARRES1 over expression elevates the levels of the antioxidant enzyme, catalase. Our results also indicate that RARRES1 expression inhibits angiogenesis in endothelial cells. CONCLUSIONS: In summary, the data presented here indicate that forced expression of RARRES1 in PCa cells (a) induces ER stress and autophagic response; (b) increases SIRT1 levels; and (c) higher levels of anti-oxidant enzymes. Our study also implicates the role of RARRES1 as a novel anti-angiogenic molecule. Overall this study reports the molecular players that RARRES1 modulates to serve as a tumor suppressor molecule. Future studies will help determine the in vivo mechanisms by which RARRES1 may serve as a target for therapeutic intervention both in cancer and in angiogenesis-related disorders.

Detection of Helicobacter felis in a cat with gastric disease in laboratory animal facility.

A 3-month-old male cat in the animal facility was presented for investigation of anorexia and occasional vomiting. We collected the specimens from gastroscopic biopsy and stool collection. The gastroscopic biopsy specimens were tested using a rapid urease test, CLO Helicobacter-detection kits. Stool specimens were gathered and evaluated using the commercially available SD Bioline H. pylori Ag kit according to the manufacturer's instructions. Genomic DNAs from gastroscopic biopsy and stool specimens of the cat were extracted and submitted to the consensus PCR to amplify Helicobacter rpoB gene. Then the DNAs from gastroscopic biopsy and stool specimens were conducted a multiplex species-specific PCR to amplify urease B gene for H. heilmannii, H. pylori and H. felis. As the results, the rapid urease test with gastroscopic biopsy was revealed positive reaction. The result of H. pylori Stool Ag assay was one red line, negative for H. pylori. The gastroscopic biopsy and stool specimen were positive reactions by the consensus PCR reaction using the RNA polymerase beta-subunit-coding gene (rpoB) to detect Helicobacter species. By multiplex species-specific PCR with gastroscopic biopsy and stool specimens, no amplification products corresponding to either H. heilmannii or H. pylori were detected, but the specimens tested were positive for H. felis. This case was confirmed as gastroenteric disease induced by H. felis infection. On our knowledge, this is a very rare report about H. felis-induced gastroenteric disease in cat and may provide a valuable data on the study of feline Helicobacter infection.

A Dunnione Compound MB12662 Improves Cisplatin-Induced Tissue Injury and Emesis.

The present study was aimed to investigate the effects of MB12662, a synthetic dunnione compound, on cisplatin-induced vomiting reflexes and intestinal, renal, immune system, and hematopoietic toxicities in ferrets and mice, respectively. Male ICR mice were orally administered MB12662 (5, 10, 25 or 50 mg/kg) for 10 days, during which intraperitoneally challenged with cisplatin (3.5 mg/kg) from day 4 to 7, and sacrificed on day 10 for the pathological examination. Male ferrets were orally administered MB12662 (25, 50 or 100 mg/kg) for 7 days, subcutaneously challenged with cisplatin (5 mg/kg), and monitored for vomiting reflexes and survival of the animals. Four-day injection of cisplatin (3.5 mg/kg) to mice caused body weight loss and degeneration and atrophy of intestinal villi, reducing villi/crypt ratio to a half level of control animals. Cisplatin also induced renal and hepatic toxicities, and depletion of splenocytes and bone marrow progenitor cells. The systemic toxicities including decreased villi/crypt ratio, immune system atrophy, splenocyte depletion, and decreased cellularity in bone marrow were improved by MB12662. Cisplatin (5 mg/kg) induced retching and emetic responses of ferrets, which were remarkably attenuated by MB12662 in a dose-dependent manner. All the ferrets pretreated with MB12662 survived the challenge of cisplatin, in comparison with 40% mortality in vehicle-treated animals, and blood parameters of nephrotoxicity and hepatotoxicity were markedly recovered. It is expected that MB12662 could be a candidate for the body protection against burden, including emesis, of chemotherapeutic agents.

Comparison of three diagnostic assays for the identification of Helicobacter spp. in laboratory dogs.

A number of Helicobacter species may confound experimental data because of their association with disease progressing in various kinds of laboratory animals. Screening of Helicobacter species is particularly desirable, because they are prevalent in commercial and research animal facilities. The aim of the present study was to compare three diagnostic methods [e.g. Helicobacter stool antigen kit (HpSA), polymerase chain reaction (PCR) and rapid urease test (RUT)] for the identification of Helicobacter spp. in stools or gastric biopsy specimens collected from eight dogs suffering from gastritis. The gastroscopic biopsy specimens were tested using RUT and PCR, while stool specimens were evaluated using both HpSA and PCR. DNAs from the gastric biopsies and stool specimens were analyzed by both a consensus PCR that amplified the RNA polymerase beta-subunit-coding gene (rpoB) of Helicobacter spp. and a species-specific PCR to amplify the urease B gene of Helicobacter heilmannii, Helicobacter pylori, and Helicobacter felis. Helicobacter spp. were detected in 62.5% of the dogs, while H. heilmannii and H. felis were identified in 37.5 and 25% of the dogs, respectively. The HpSA did not efficiently detect Helicobacter spp. in the stool samples compared to the RUT and PCR assays, both of which successfully detected Helicobacter spp. in the two sample types. Finally, we recommend that consensus PCR with stool specimens could be used before the species-specific PCR for identifying Helicobacter species in laboratory dogs.

Onion peel water extracts enhance immune status in forced swimming rat model.

Onion peel contains a high concentration of quercetin and other flavonoids. In this study, the potential immune-enhancing effects of an onion peel water extract (OPE) supplement were investigated by the rat forced swimming test. OPE was prepared using hot water. Thirty-six male Sprague Dawley rats were fed a pellet diet for 1 week and were then randomly divided into six groups: normal control (NC), forced swimming control (FSC), positive control (quercetin 20 mg/kg), and three groups administered 4, 20, or 100 mg/kg of OPE. Oral drug administration was conducted daily for 4 weeks. All rats, except those of NC group, were forced to swim in water and were considered exhausted when they failed to rise to the water surface to breathe within a 7-s period. Blood lymphocyte counts, immune organ weights, histopathological analysis, and serum interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and interleukin (IL)-12 levels were determined. OPE-treated rats consumed more food and had an increased thymic cortex to medulla ratio than that observed in FSC group rats (P<0.05). The area of the white pulp in the spleens of OPE-treated group rats was increased compared with that in FSC group rats (P<0.05). Furthermore, blood lymphocyte numbers and IFN-γ, TNF-α, and IL-12 concentrations were significantly higher in OPE-fed groups than in FSC group (P<0.05). These results suggest that an OPE supplement can improve the immune status by increasing the number of immune-related cells and specific cytokine levels.

Immunostimulating effects of extract of Acanthopanax sessiliflorus.

As malfunction/absence of immune cells causes a variety of immunosuppressive disorders and chemical synthetic drugs for curing these diseases have many adverse effects, vigorous studies are being conducted. The Acanthopanax family has been used as traditional medicines for gastric ulcer, diabetes, etc. and culinary materials in East-South Asia. In this study, the immunostimulating properties of A. sessiliflorus were evaluated. A. sessiliflorus increased not only the splenocyte number but also immune-related cytokines such as TNF-α. However, it could not upregulate the expressions of IFN-γ and IL-2. A. sessiliflorus increased the swimming time, and comparison of organ weights relative to body weights for immune-related organs such as the spleen and thymus after a forced swim test showed that it could recover the spleen and thymus weights. It also increased the expression of TNF-α and slightly increased the concentration of IFN-γ but not IL-2. From the results, we concluded that as A. sessiliflorus has not only a host defense effect but also a stress-ameliorating property, further study it will be a promising material of immunostimulating material.

Usefulness of a Helicobacter pylori stool antigen test for diagnosing H. pylori infected C57BL/6 mice.

Among several diagnostic tests, a Helicobacter pylori stool antigen (HpSA) test may offer a useful noninvasive method for diagnosing infection without sacrificing animals. In this study, male C57BL/6 mice (n=6) were infected with H. pylori ATCC 49503 (1×10(8) CFU/mouse) by intragastric inoculation three times at 2-day intervals, and H. pylori infected stool specimens were collected 1, 3, 5, 7, 14, 21 days after infection to assess reliability of the HpSA test. Five of six specimens were positive at 5-21 days after infection, and the sensitivity of the HpSA test was 83.33%. The presence of H. pylori infection was confirmed by the rapid urease test and genomic DNA polymerase chain reaction (PCR), and showed the same results as the HpSA. However, the rapid urease test and genomic DNA PCR are invasive tests and require animal sacrifice to detect H. pylori in gastric biopsy samples. We suggest that an HpSA test kit would be useful and effective for monitoring H. pylori in various laboratory animals, as H. pylori can be easily monitored without sacrificing animals.

Protective effects of the ethanolic extract of Melia toosendan fruit against colon cancer.

Colorectal cancer is one of the leading causes of death in the world. Plant-derived products have proven to be valuable sources for discovery and development of unique anticancer drugs. In this study, the inhibitory effects of ethanolic extract of Melia toosendan fruit (EMTF), a traditional medicine in the Chinese Pharmacopeia were evaluated in vitro and in vivo against colon cancer. Human colon cancer cells SW480 and murine colorectal adenocarcinoma cells CT26 were used to investigate cell proliferation. The results showed that EMTF inhibited cell proliferation of SW480 and CT26 by promoting apoptosis as indicated by nuclear chromatin condensation and DNA fragmentation. Through increasing mitochondrial membrane permeability and cytochrome c release from mitochondria, EMTF induced caspase-9 activity which further activated caspase-3 and poly(ADP-ribose) polymerase cleavage, leading the tumor cells to apoptosis. The in vivo results confirmed reduction of tumor volume and apoptotic effects and the side effects were not induced by EMTF. Therefore, EMTF may be an effective chemotherapeutic agent for colon cancer treatment.

Spontaneous osteosarcoma of the femur in a non-obese diabetic mouse.

An abnormal swelling was identified in the distal portion of the right femur in a 1-year-old non-obese diabetic (NOD) mouse. Grossly, a large mass of the distal femur was observed in the right femur. Lesions were poorly marginated, associated with destruction of the cancellous and cortical elements of the bone, and showed ossification within the soft tissue component. Histologically, the tumor was identified as a poorly differentiated sarcoma. Histopathologic examination of the bone masses revealed invasive proliferation of poorly differentiated neoplastic mesenchymal cells forming streams, bundles, and nests, which resulted in destruction of normal bone. Neoplastic cells exhibited random variation in cellular appearance and arrangement, as well as matrix composition and abundance. Haphazard and often intermingling patterns of osteogenic, chondroblastic, lipoblastic, and angiogenic tissues were present. Larger areas of neoplastic bone and hyaline cartilage contained multiple large areas of hemorrhage and necrosis bordered by neoplastic cells. The mass was diagnosed as an osteosarcoma. To our knowledge, this is the first spontaneous osteosarcoma in an NOD mouse.

Spontaneous sertoli cell tumor with cryptorchism in a beagle dog.

A male one year-old beagle dog with unilateral cryptorchism was presented for investigation of reduced appetite. Abdominal sonography and radiography demonstrated abnormal enlargement of the left testicle in the abdominal cavity. Both the retroperitoneal cryptorchid testicle and the other contralateral testicle were removed surgically. The retroperitoneal cryptorchid testicle was an enlarged, firm and bulging sphere mass. The cut surface revealed a homogeneous white color. The contralateral testicle in the scrotum showed an almost normal appearance. Histopathologically, the retroperitoneal cryptorchid testicle was diagnosed as a Sertoli cell tumor. This report describes a case of Sertoli cell tumor with cryptorchism in a beagle dog.

Uterine adenocarcinoma with feline leukemia virus infection.

Feline endometrial adenocarcinomas are uncommon malignant neoplasms that have been poorly characterized to date. In this study, we describe a uterine adenocarcinoma in a Persian cat with feline leukemia virus infection. At the time of presentation, the cat, a female Persian chinchilla, was 2 years old. The cat underwent surgical ovariohystectomy. A cross-section of the uterine wall revealed a thickened uterine horn. The cat tested positive for feline leukemia virus as detected by polymerase chain reaction. Histopathological examination revealed uterine adenocarcinoma that had metastasized to the omentum, resulting in thickening and the formation of inflammatory lesions. Based on the histopathological findings, this case was diagnosed as a uterine adenocarcinoma with abdominal metastasis. To the best of our knowledge, this is the first report of a uterine adenocarcinoma with feline leukemia virus infection.

A single mutation at lysine 241 alters expression and trafficking of the D2 dopamine receptor.

Ubiquitination of G protein-coupled receptors has been identified to regulate receptor signal transduction including agonist-induced internalization and sorting of internalized receptor for degradation or for recycling. Using co-immunoprecipitation and immunoblot analysis, I found that the membrane-associated D(2) dopamine receptor (DAR) is mono-ubiquitinated in the absence of an agonist following heterologous expression in human embryonic kidney cells (HEK293). By using site-directed mutagenesis, this report shows that the loss of lysine-241, K241A D(2) DAR reduced the amount of membrane-associated D(2) DAR. It is of interest that the K241A D(2) DAR also had a distinctly different ubiquitination pattern than the wild-type D(2) DAR. It is important to note that the ubiquitinated mutant D(2) DAR was degraded through ubiquitin-proteasome pathway. These data provide the factual evidence that a loss of lysine-241 of the D(2) DAR affects receptor ubiquitination and renders the protein susceptible to the proteasomal degradation.

Malignant catarrhal fever-like disease in sheep after intranasal inoculation with ovine herpesvirus-2.

A malignant catarrhal fever (MCF)-like disease was induced experimentally in 3 sheep after aerosol inoculation with ovine herpesvirus-2 (OvHV-2). Each of 3 OvHV-2-negative sheep was nebulized with 2 ml of nasal secretions containing approximately 3.07 X 10(9) OvHV-2 DNA copies from a sheep experiencing an intensive viral-shedding episode. Ovine herpesvirus-2 DNA became detectable by polymerase chain reaction in the peripheral blood leukocytes of all 3 sheep within 3 days, and all 3 seroconverted between 6 and 8 days postinfection (PI). The sheep developed clinical signs, with copious mucopurulent nasal discharge and fever around 14 days PI. One of the 3 clinically affected sheep was euthanized at 18 days PI. Major lesions at necropsy were multifocal linear erosions and ulcers in mucosa of the cheeks, tongue, pharynx, and proximal esophagus and mild disseminated pneumonia. Microscopically, there was extensive moderate superficial histiocytic-lymphocytic rhinitis with epithelial dissociation and degeneration. Moderate multifocal histiocytic bronchointerstitial pneumonia was associated with loss of terminal bronchiolar epithelium. Lymphocytic vasculitis was present only in the lung. The remaining 2 sheep recovered clinically, approximately 25 days PI. The study revealed that clinical signs and lesions resembling MCF can develop when uninfected sheep are exposed to a high dose of aerosolized OvHV-2.

Cystic endometrial hyperplasia and endometritis in a dog following prolonged treatment of medroxyprogesterone acetate.

An 8-year-old female Yorkshire Terrier was presented for investigation of reduced appetite, and occasional vomiting. She has been treated with medroxyprogesterone acetate (MPA) from past 3 year-old age for contraception. Abdominal sonography showed abnormal enlargement of uterus, and ovariohysterectomy was performed. Main gross findings of uterus were enlarged lesions in two areas of the left horn, which had thickened wall and yellowish sticky material in the lumen. Histopathologically, cystic endometrial hyperplasia (CEH) and endometritis were present in the thickened area. In this case, CEH and endometritis may be attributed to prolonged treatment of MPA. It was concluded that further study is needed to clarify the association of MPA treatment with age, its pathogenesis and abnormal uterine changes in dogs.