Antioxidative and Anti-photooxidative Potential of Interruptins from the Edible Fern Cyclosorus terminans in Human Skin Cells.

Background: Human skin is exposed daily to oxidative stress factors such as UV light, chemical pollutants, and invading organisms. Reactive oxygen species (ROS) are intermediate molecules that cause cellular oxidative stress. In order to survive in an oxygen-rich environment, all aerobic organisms, including mammals, have evolved enzymatic and non-enzymatic defence systems. The interruptins from an edible fern Cyclosorus terminans possess antioxidative properties and can scavenge intracellular ROS in adipose-derived stem cells.

Objectives: This study aimed to evaluate the antioxidative efficacy of interruptins A, B, and C in cultured human dermal fibroblasts (HDFs) and epidermal keratinocytes (HEKs). Moreover, the anti-photooxidative activity of interruptins in ultraviolet (UV)-exposed skin cells was investigated.

Methods: The intracellular ROS scavenging capacity of interruptins in skin cells was measured by flow cytometry. Their induction effects on gene expression of the endogenous antioxidant enzymes was monitored using real-time polymerase chain reaction.

Results: Interruptins A and B, but not interruptin C, were highly effective in ROS scavenging, particularly in HDFs. Interruptins A and B upregulated gene expression of superoxide dismutase (SOD)1, SOD2, catalase (CAT), and glutathione peroxidase (GPx) in HEKs, but they only induced SOD1, SOD2, and GPx gene expression in HDFs. Additionally, interruptins A and B efficiently suppressed UVA- and UVB-induced ROS generation in both HEKs and HDFs.

Conclusion: The results suggest that these naturally occurring interruptins A and B are potent natural antioxidants and therefore may have the potential in the future of inclusion in antiaging cosmeceutical products.

Anti-cancer effect of engineered recombinant interleukin 18.

BACKGROUND: Interleukin 18 (IL-18) is an inflammatory cytokine belonging to the interleukin 1 (IL-1) superfamily, and is known for its role in anti-cancer activity by promoting type 1 immune response, and thus may be applied to cancer immunotherapy. Our previous report has showed 16 times higher activity of engineered E6K+T63A IL-18 than of native IL-18 in vitro. However, no data has been acquired for its anti-cancer effect in animal model. OBJECTIVES: To investigate the anti-cancer effect of engineered E6K+T63A IL-18 as an immune stimulant in vivo. MATERIAL AND METHODS: Tumor-bearing mice were treated with native IL-18 or E6K or E6K+T63A IL-18 once a day for 10 days after the tumor reached the volume of 100 mm3. Tumor volume and the number of certain immune cell type in the tumor microenvironment were investigated in this study. RESULTS: The results showed that tumor progression in mice treated with E6K+T63A was slower than in mice treated with E6K and native IL-18. The volume of the tumor was also smaller and the lifespan longer in the E6K+T63A IL-18-treated mice. The proportions of type 1 helper T cell (Th1) and cytotoxic T lymphocyte (CTL) were significantly higher in mice treated with E6K+T63A IL-18. CONCLUSIONS: These results suggest that our engineered IL-18 conferred strong anti-tumor immunity in the animal model.

Investigation of Leptospira Infection in Stray Animals in Songkhla, Thailand: Leptospirosis Risk Reduction in Human.

Leptospirosis is a zoonosis of public health concern in Thailand. Human leptospirosis presents severe illness and can be fatal due to pulmonary hemorrhage, kidney failure, or cardiac impairment. Infected animals show no clinical signs and play an important role in the Leptospira infection of humans and other hosts. The prevalence of leptospirosis in stray animals in Thailand is unknown. The aim of this study was to investigate the incidence of Leptospira infection in stray animals including dogs and cats in Songkhla province, Thailand. A total of 434 blood samples were collected from 370 stray dogs and 64 stray cats during a population control program from 2014 to 2018. Screening the serum samples using the latex agglutination test to detect antibodies against Leptospira interrogans showed that 29.26% (127/434) were positive. There were 120 positive samples for stray dogs and 7 positive samples for stray cats. The detection of positive samples by polymerase chain reaction specific to the LipL32 of L. interrogans showed 1.61% (7/434) were positive. Stray cats (5/64) showed a higher prevalence than stray dogs (2/370), which might be because they are more likely to come into contact with rodents in their habitat. Although the active infection detected was low, the seroprevalence was high. This result indicated that the stray animals might not have been infected at the time of sample collection, but that they had been infected in the past or were in a latent period of infection. Therefore, they might serve as a leptospirosis reservoir for domestic animals and humans present in the same environment. The results show that stray animals need health care, such as vaccination, surveillance, and treatment, when infected to prevent or reduce the risk of transmission to humans.

Investigation of Melioidosis Outbreak in Pig Farms in Southern Thailand.

Melioidosis, caused by the Gram-negative bacterium Burkholderia pseudomallei, is a potentially life-threatening infection that can affect humans and a wide variety of animals in the tropics. In December 2017, a swine melioidosis case was discovered during a meat inspection at a privately-owned slaughterhouse in Nakhon Si Thammarat Province in southern Thailand. The infection, which continued for several months, caused a dispute about where the disease began. An environmental investigation into two farms-both involved in raising the first infected pig-ensued. Through genetic analysis, the investigation revealed that a contaminated water supply at one farm was the probable source of infection. The three local sequence types identified in the investigation were types 51, 298 and 392.

Successful Treatments and Management of A Case of Canine Melioidosis.

This communication presents a successful story of an attempt to treat and manage a case of canine melioidosis, a severe tropical disease caused by Burkholderia pseudomallei. A 10-year-old dog was trapped with barbed wires, causing an infected wound around its neck and back, which was later diagnosed as severe melioidosis. The dog was treated based on a modified human protocol. Intravenous meropenem injections (20 mg/kg twice daily) were given for 14 days to prevent death from sepsis prior to treatment with oral sulfamethoxazole-trimethoprim (25 mg/kg twice daily) for 20 weeks to eliminate the bacteria. Canine melioidosis is an unusual infection in dogs, even in Thailand where melioidosis is highly endemic. This successful case management was solely based on proper diagnosis and appropriate treatments.

miR-144 regulates oxidative stress tolerance of thalassemic erythroid cell via targeting NRF2.

Thalassemia has a high prevalence in Thailand. Oxidative damage to erythroid cells is known to be one of the major etiologies in thalassemia pathophysiology. Oxidative stress status of thalassemia is potentiated by the heme, nonheme iron, and free iron resulting from imbalanced globin synthesis. In addition, levels of antioxidant proteins are reduced in α-thalassemia and ß-thalassemia erythrocytes. However, the primary molecular mechanism for this phenotype remains unknown. Our study showed a high expression of miR-144 in ß- and α-thalassemia. An increased miR-144 expression leads to decreased expression of nuclear factor erythroid 2-related factor 2 (NRF2) target, especially in α-thalassemia. In α-thalassemia, miR-144 and NRF2 target are associated with glutathione level and anemia severity. To study the effect of miR-144 expression, the gain-loss of miR-144 expression was performed by miR inhibitor and mimic transfection in the erythroblastic cell line. This study reveals that miR-144 expression was upregulated, whereas NRF2 expression and glutathione levels were decreased in comparison with the untreated condition after miR mimic transfection, while the reduction of miR-144 expression contributed to the increased NRF2 expression and glutathione level compared with the untreated condition after miR inhibitor transfection. Moreover, miR-144 overexpression leads to significantly increased sensitivity to oxidative stress at indicated concentrations of hydrogen peroxide (H2O2) and rescued by miR-144 inhibitor. Taken together, our findings suggest that dysregulation of miR-144 may play a role in the reduced ability of erythrocyte to deal with oxidative stress and increased RBC hemolysis susceptibility especially in thalassemia.

Seropositivities against brucellosis, coxiellosis, and toxoplasmosis and associated factors in pregnant women with adverse pregnancy outcomes: A cross-sectional study.

BACKGROUND: Brucellosis, coxiellosis, and toxoplasmosis can be transmitted from infected ruminants to pregnant women and may induce adverse pregnancy outcomes; however, there are to date few studies. This study aimed to examine the seropositivities of immunoglobulin G (IgG) against those three pathogens among pregnant women with adverse pregnancy outcomes, and to explore the associated factors. METHODS: A cross-sectional study was conducted in southern Thailand, where goat production is common. A total of 105 pregnant Thai women who had adverse pregnancy outcomes and serum samples collected at first antenatal care visit before their 28th gestational week from June 2015 to June 2016 were included. The seropositivities of IgG anti-Brucella abortus, Toxoplasma gondii, and Coxiella burnetii antibodies were tested by using commercial enzyme-linked immunosorbent assay (ELISA) kits. Associated factors with seropositivity were analyzed using multiple logistic regression. RESULTS: Most women were Muslim aged 20-34 years and 32.4% had a prior history of one or more adverse pregnancy outcomes. One-third of the women had been exposed to goats or raw goat products. Of the 105 serum samples, the seropositivity of anti-T. gondii IgG was highest (33/105, 31.4%), followed by anti-C. burnetii IgG (2/105, 1.9%), and anti-B. abortus IgG (1/105, 1.0%), respectively. None of the pregnant women were found to be co-seropositive for those three pathogens. CONCLUSIONS: One-third of women with adverse pregnancy outcomes showed positive antibodies for toxoplasmosis, coxiellosis and brucellosis. A dose-response relationship between seropositivity of anti-T. gondii IgG and age was noticed.

Risk of adverse pregnancy outcomes and seroprevalence for brucellosis in pregnant women exposed to goats or raw goat products in southern Thailand: a prospective cohort study.

BACKGROUND: Pregnant women infected with brucellosis have been shown to have higher odds of having been exposed to goats and raw goat products and adverse pregnancy outcomes. However, information on these associations in asymptomatic pregnant women is limited, particularly in the brucellosis-endemic areas. This study aimed to assess the association of a history of exposure to goats and/or raw goat products and the serological status of anti-Brucella abortus immunoglobulin G (IgG) with adverse pregnancy outcomes among pregnant women, and explore factors associated with having exposure to goats and/or raw goat products. METHODS: A prospective cohort study was conducted among pregnant women from July 2015 to July 2016 at Songkhla province in southern Thailand. All pregnant women who came for antenatal care (ANC) visits were approached. Blood samples from the women who agreed to participate were randomly tested for anti-Brucella abortus IgG. The women were then followed for adverse pregnancy outcomes. RESULTS: Of 666 pregnant women, the majority (74.4%) were aged 20-34 years and Muslim (89.2%), 30.6% indicated exposure to goats or raw goat products, and 17.3% had adverse pregnancy outcomes. Women rearing goats at home or having neighbors rearing goats were more likely to be exposed to goats or raw goat products by cutaneous contact. Of 465 women having a blood test, 3.7% had seropositive results for anti-Brucella abortus IgG. No association with adverse pregnancy outcomes was found in the women reporting any exposure to goat and raw goat products. Having the first ANC visit at the first trimester and history of preterm birth or low birth weight newborn were independent risk factors of adverse pregnancy outcomes. Women who had positive serological results were more likely to have a history of drinking raw goat milk than those who had negative results significantly. CONCLUSIONS: Although no association between past exposure with goats and raw goat products and adverse pregnancy outcomes was found, women with past exposure showed positive anti-Brucella abortus IgG. Counseling on avoiding consumption of raw goat milk would be beneficial to prevent goat-related infection in pregnant women in this area.

Rhodomyrtone as a potential anti-proliferative and apoptosis inducing agent in HaCaT keratinocyte cells.

Psoriasis is a skin disease associated with hyperproliferation and abnormal differentiation of keratinocytes. Available approaches using synthetic drugs for the treatment of severe psoriasis may cause side effects. Alternatively, plant-derived compounds are now receiving much attention as alternative candidates for the treatment of psoriasis. In this study, the effects of rhodomyrtone, a bioactive plant extract isolated from Rhodomyrtus tomentosa leaves on the proliferation, growth arrest, and apoptosis of HaCaT keratinocytes were investigated. Percentage anti-proliferative activity of rhodomyrtone on HaCaT cells at concentrations of 2-32µg/ml after 24, 48, and 72h ranged from 13.62-61.61%, 50.59-80.16%, and 61.82-85.34%, respectively. In a scratch assay, rhodomyrtone at 2 and 4µg/ml significantly delayed closure of a wound by up to 61.78%, and 71.65%, respectively, after 24h incubation. HaCaT keratinocytes treated with rhodomyrtone showed chromatin condensation and fragmentation of nuclei when stained with Hoechst 33342. This indicated that rhodomyrtone induced apoptosis in the keratinocytes. In addition, flow cytometric analysis demonstrated an increase in the percentage of apoptosis of keratinocytes after treatment with rhodomyrtone at 2-32µg/ml from 1.2-10%, 8.2-35.4%, and 21.0-77.8% after 24, 48, and 72h, respectively, compared with the control. To further develop the compound as a potential anti-psoriasis agent, a rhodomyrtone formulation was prepared and subjected to skin irritation tests in rabbits. The formulation caused no skin irritation including such as erythema and edema. The results indicated that rhodomyrtone had the potential as a promising candidate for further development as a natural anti-psoriasis agent.

A prospective evaluation of four immunodiagnotic assays for human leptospirosis.

This was a prospective evaluation of four immunodiagnostic assays for human leptospirosis, including the indirect immunofluorescence test (IFA), the microscopic agglutination test (MAT), the LEPTO dipstick, and the latex agglutination (LA) test. Four hundred ninety-two serum samples were collected from 348 patients who presented with acute fever without localizing signs. The sensitivities of the IFA, MAT, Dipstick, and LA were 91.9, 76.6, 77.4, and 83.1%, respectively. The specificities of these assays were 100.0, 100.0, 89.3, and 83.5, respectively. Even though IFA showed the highest overall sensitivity and specificity, when acute sera were considered, the LA was the most sensitive (28.7%). All 3 genus specific antibody assays had broad reactivity against various serogroups. The MAT is best suited for the reference laboratory, where it can be maintained with the battery of live antigens; the IFA is suited for a laboratory with sophisticated equipment and technical expertise; the Dipstick and LA are suitable for peripheral laboratories which lack expensive equipment and expertise.