Reduction of carcinogens in fermented fish (pla-ra and pla-som) by heating.

Background and Aim: The risk factors for cholangiocarcinoma (CCA) are opisthorchiasis and the intake of a combination of nitroso compounds through the consumption of traditionally fermented fish, which is very popular in areas where liver flukes are endemic. The incidence of CCA remains high because this cultural habit of rural people has been altered. Therefore, decreasing nitrate and nitrite concentrations in fermented fish are an alternative approach to reducing the risk of CCA. Thus, this study aimed to reduce nitrate and nitrite concentrations in fermented foods by heating and investigated its effect on CCA development in a hamster model. Materials and Methods: We used Association of Official Analytical Chemists method 973.31 to measure the nitrate and nitrite concentrations in both fermented fish (pla-ra [PR]) and pickled fish (pla-som [PS]) before and after boiling for 5 and 30 min, respectively. The same samples were fed to Opisthorchis viverrini (OV)-infected or -uninfected hamsters for 3 months. Thereafter, the hamsters' liver and blood were collected for analysis. Results: The levels of nitrates and nitrites in PS and PR significantly decreased following boiling for 5 and 30 min. The OV-PR and OV-PS groups showed dramatically increased numbers of inflammatory cells, fibrosis surrounding the bile duct, and focal fibrotic areas. However, after boiling the fermented dishes for 5 and 30 min, the extent of inflammatory cell infiltration and intensity of fibrosis in these groups were decreased. Conclusion: Our findings suggest that boiling reduces nitrate and nitrite toxicity in fermented dishes, as evidenced by reduced hepatic inflammation. However, regardless of heating, kidney tissues are adversely affected when fermented meals are consumed daily.

Acetaminophen Overdose Enhances Early Cholangiocarcinoma in Opisthorchiasis Hamsters.

Opisthorchiasis which exerted by infection of Opisthorchis viverrini is strongly related to the incident of cholangiocarcinoma (CCA) in many Southeast Asian countries northeastern of Thailand. The O. viverrini infection is primarily caused by raw fish consumption, and repeated exposure to liver fluke. Meanwhile, acetaminophen is usually medicated to relieve pain in particularly people in northeast Thailand. OBJECTIVE: This study therefore aimed at investigating effects of acetaminophen on pathogenesis in hamsters for opisthorchiasis. METHODS: There were 4 groups of hamsters: i) uninfected hamster (N); ii) sole acetaminophen administration (N-Ac); iii) sole O. viverrini infection (OV); and iv) combination of O. viverrini infection and acetaminophen (OV-Ac) on pathology of hamsters for 1 month post infection. For analysis of histopathological changes through hematoxylin and eosin, Sirius red and immunohistostaining for Cytokeratin 19 (CK-19), Proliferating cell nuclear antigen (PCNA) and CA 19-9, serum's hamsters were used detected for liver function tests and tumor-related genes expression. RESULTS: After 1 month under these treatments, the OV-Ac showed significantly higher CCA risk, including inflammatory cells were aggregations around bile duct, new bile duct and fibrosis in subcapsular hepatic tissues, than other treatments. These pathological parameters were positively correlated with immunohistochemical staining derived from CK-19, PCNA and CA 19-9. In addition, OV-Ac had significantly higher liver function tests (ALT). CONCLUSION: Combined intake of liver fluke-contaminated raw fishes and acetaminophen rendered more severity of CCA than sole consumption of the contaminated raw fishes.

In Vivo Efficacy of Clove Essential Oil Ointment for Microsporum gallinae Avian Dermatophytosis-A Randomized Controlled Trial.

Avian favus (dermatophytosis) is a superficial mycosis caused by Microsporum gallinae in poultry. This disease is an important problem in poultry husbandry, but the standard antifungal treatment can leave drug residues in farm products. The aim of this study was to compare the efficacy of a clove essential oil ointment (3%, w/w) with commercially available ketoconazole cream (2%, w/w) for the treatment of M. gallinae infection in chickens. An in vitro time-kill assay showed that clove essential oil ointment reduced the number of viable M. gallinae ATCC 90749 by 99.99% within 1 hr. A randomized controlled trial showed that the therapeutic efficacy of clove essential oil ointment (3%, w/w) was noninferior to ketoconazole cream (2%, w/w) in M. gallinae-infected chickens. The percentage of completely recovered (culture-negative) animals in both treatment groups was 90% in day 35 after initial treatment. This study indicates that clove essential oil is suitable for preparation as an alternative topical treatment for avian dermatophytosis.

Eficacia in vivo de una pomada de aceite esencial de clavo para la dermatofitosis aviar por Microsporum gallinae: Ensayo controlado y aleatorio. El favus aviar (dermatofitosis) es una micosis superficial causada por Microsporum gallinae en la avicultura. Esta enfermedad es un problema importante en la avicultura, pero el tratamiento antifúngico estándar puede dejar residuos de medicamentos en los productos agrícolas. El objetivo de este estudio fue comparar la eficacia de una pomada de aceite esencial de clavo (3%, peso a peso; P/P) con la crema de ketoconazol disponible comercialmente (2%, P/P) para el tratamiento de la infección por M. gallinae en pollos. Un ensayo in vitro para determinar el tiempo de eliminación mostró que la pomada de aceite esencial de clavo redujo el número de M. gallinae ATCC 90749 viables en un 99.99% en una hora. Un ensayo controlado y aleatorio mostró que la eficacia terapéutica de la pomada de aceite esencial de clavo (3%, P/P) no fue inferior a la crema de ketoconazol (2%, P/P) en pollos infectados con M. gallinae. El porcentaje de animales completamente recuperados (con cultivo negativo) en ambos grupos de tratamiento fue del 90% en el día 35 después del tratamiento inicial. Este estudio indica que el aceite esencial de clavo es adecuado para su preparación como tratamiento tópico alternativo para la dermatofitosis aviar.

DEVELOPMENT OF AN ANTIBIOTIC OPTIONS INDEX FOR ANTIBIOTIC RESISTANCE MONITORING.

Using antibiogram data to indicate the overall antibiotic resistance of a pathogen is complicated by the multiple antibiotic susceptibilities reported in the antibiogram. The objectives of this study were to develop and determine the benefits of an Antibiotic Options Index (AOI); an index that summarizes antibiotic susceptibility data for a pathogen by presenting it as the availability of antibiotic treatment options. The AOI was calculated using antibiogram data for the seven most commonly isolated pathogens from the National Antimicrobial Resistance Surveillance Center of Thailand between 1998 and 2014 and was classified as acceptable (AOI ≥ 0.8) or unacceptable (AOI < 0.8) based on the availability of treatment options. The AOI identified two problematic pathogens: Acinetobacter baumannii and methicillin-resistant Staphylococcus aureus (MRSA). For A. baumannii, the probability of having at least two viable antibiotic treatment options (AOIm2) decreased from an acceptable level (0.93) in 1998 to an unacceptable level (0.53) in 2014 and for MRSA the AOIm2 decreased from an acceptable level (0.82) in 1998 to an unacceptable level (0.47) in 2014. By including the idea that the problem with increasing antibiotic resistance is a problem with treating infections, the AOI effectively compiles susceptibility data to present it as the probability of having effective antibiotic treatment. This index is calculated from widely available antibiogram data, making it more suitable to be used to monitor antibiotic resistance at the hospital, provincial and national levels.

A role for altered phagosome maturation in the long-term persistence of Helicobacter pylori infection.

The vigorous host immune response that is mounted against Helicobacter pylori is unable to eliminate this pathogenic bacterium from its niche in the human gastric mucosa. This results in chronic inflammation, which can develop into gastric or duodenal ulcers in 10% of infected individuals and gastric cancer in 1% of infections. The determinants for these more severe pathologies include host (e.g., high IL-1ß expression polymorphisms), bacterial [e.g., cytotoxicity-associated gene (cag) pathogenicity island], and environmental (e.g., dietary nitrites) factors. However, it is the failure of host immune effector cells to eliminate H. pylori that underlies its persistence and the subsequent H. pylori-associated disease. Here we discuss the mechanisms used by H. pylori to survive the host immune response and, in particular, the role played by altered phagosome maturation.

Helicobacter pylori phagosome maturation in primary human macrophages.

BACKGROUND: Helicobacter pylori (H. pylori) is a micro-aerophilic, spiral-shaped, motile bacterium that is the principal cause of gastric and duodenal ulcers in humans and is a major risk factor for the development of gastric cancer. Despite provoking a strong innate and adaptive immune response in the host, H. pylori persists in the gastric mucosa, avoiding eradication by macrophages and other phagocytic cells, which are recruited to the site of infection. Here we have characterised the critical degradative process of phagosome maturation in primary human macrophages for five genotypically and phenotypically distinct clinical strains of H. pylori. RESULTS: All of the H. pylori strains examined showed some disruption to the phagosome maturation process, when compared to control E. coli. The early endosome marker EEA1 and late endosome marker Rab7 were retained on H. pylori phagosomes, while the late endosome-lysosome markers CD63, LAMP-1 and LAMP-2 were acquired in an apparently normal manner. Acquisition of EEA1 by H. pylori phagosomes appeared to occur by two distinct, strain specific modes. H. pylori strains that were negative for the cancer associated virulence factor CagA were detected in phagosomes that recruited large amounts of EEA1 relative to Rab5, compared to CagA positive strains. There were also strain specific differences in the timing of Rab7 acquisition which correlated with differences in the rate of intracellular trafficking of phagosomes and the timing of megasome formation. Megasomes were observed for all of the H. pylori strains examined. CONCLUSIONS: H. pylori appeared to disrupt the normal process of phagosome maturation in primary human macrophages, appearing to block endosome fission. This resulted in the formation of a hybrid phagosome-endosome-lysosome compartment, which we propose has reduced degradative capacity. Reduced killing by phagocytes is consistent with the persistence of H. pylori in the host, and would contribute to the chronic stimulation of the inflammatory immune response, which underlies H. pylori-associated disease.

Lysosomal storage disease: revealing lysosomal function and physiology.

The discovery over five decades ago of the lysosome, as a degradative organelle and its dysfunction in lysosomal storage disorder patients, was both insightful and simple in concept. Here, we review some of the history and pathophysiology of lysosomal storage disorders to show how they have impacted on our knowledge of lysosomal biology. Although a significant amount of information has been accrued on the molecular genetics and biochemistry of lysosomal storage disorders, we still do not fully understand the mechanistic link between the storage material and disease pathogenesis. However, the accumulation of undegraded substrate(s) can disrupt other lysosomal degradation processes, vesicular traffic, and lysosomal biogenesis to evoke the diverse pathophysiology that is evident in this complex set of disorders.

Monocyte and macrophage killing of helicobacter pylori: relationship to bacterial virulence factors.

BACKGROUND: Helicobacter pylori infection is an important health problem, as it involves approximately 50% of the world's population, causes chronic inflammatory disease and increases the risk of gastric cancer development. H. pylori infection elicits a vigorous immune response, but this does not usually result in bacterial clearance. We have investigated whether the persistence of H. pylori in the host could be partly due to an inability of macrophages to kill this bacterium. MATERIALS AND METHODS: Monocytes and macrophages isolated from the peripheral blood of normal human controls were infected in vitro with five H. pylori isolates. The isolates were characterized for known H. pylori virulence factors; vacuolating cytotoxin (VacA), the cag pathogenicity island (cagPAI), urease, and catalase by Western blot and polymerase chain reaction analysis. The ability of primary human monocytes and macrophages to kill each of these H. pylori strains was then defined at various time points after cellular infection. RESULTS: The five H. pylori strains showed contrasting patterns of the virulence factors. There were different rates of killing for the bacterial strains. Macrophages had less capacity than monocytes to kill three H. pylori strains. There appeared to be no correlation between the virulence factors studied and differential killing in monocytes. CONCLUSIONS: Primary human monocytes had a higher capacity to kill certain strains of H. pylori when compared to macrophages. The VacA, cagPAI, urease, and catalase virulence factors were not predictive of the capacity to avoid monocyte and macrophage killing, suggesting that other factors may be important in H. pylori intracellular pathogenicity.