Morphological Changes and Inflammation Preceded the Pathogenesis of 1,2-Dimethylhydrazine-Induced Colorectal Cancer.

OBJECTIVE: This study examined the morphological changes in the colonic mucosa and the presence of inflammation in rats induced with 1,2-dimethylhydrazine (DMH) 30 mg/kg BW over 9, 11, and 13 weeks without a latency period. METHODS: Hematoxylin and eosin staining was performed to assess the morphology and characteristic alteration of the epitheliocytes in the colon. Immunohistochemistry was employed to assess the expression of tumor necrosis factor (TNF)-α and cyclooxygenase-2 (COX-2). The difference in the severity of inflammation and COX-2 expression was examined using one-way analysis of variance. The correlation of COX-2 expression with the severity of inflammation was analyzed using Spearman's rank correlation test. RESULT: Until week 13, chronic inflammation and non-hyperplastic and hyperplastic aberrant crypt foci occurred. The severity of inflammation gradually shifted from high moderate to low moderate. TNF-α expression was high in all groups; however, COX-2 expression was gradually lower with longer duration of induction, which corresponded with the severity of inflammation. CONCLUSION: DMH induction until week 13 without a latency period caused chronic inflammation without the formation of adenoma or adenocarcinoma. A very strong correlation was established between COX-2 expression and inflammation.

Tumour-suppressive effects of curcumin analogs CCA-1.1 and Pentagamavunone-1 in colon cancer: In vivo and in vitro studies.

This study aimed to evaluate the efficacy of Chemoprevention Curcumin Analog-1.1 (CCA-1.1) and Pentagamavunone-1 (PGV-1) in vivo and in vitro in colorectal cancer model. CCA-1.1 or PGV-1 was administered orally to 1,2-dimethylhydrazine (DMH)-induced rats for 16 weeks. The cytotoxicity of both compounds was tested on Caco-2, CT26, and NIH/3T3 cells using the MTT method. The cell cycle, apoptosis, and reactive oxygen species (ROS) levels were analyzed through flow cytometry. X-gal staining was used to examine the compound's effect on senescence. Oral co-administration of CCA-1.1 or PGV-1 significantly suppressed the carcinogenic characteristics and symptoms of premalignant colon cancer relative to DMH-only and untreated groups. CCA-1.1 and PGV-1 administration did not affect the blood profile. CCA-1.1 and PGV-1 demonstrated great cytotoxicity on Caco-2 and CT26 cells, with 50% inhibition concentration (IC50) values of 4.3 ± 0.2 and 3.1 ± 0.1 µM for CCA-1.1 and 11.2 ± 1.1 and 4.8 ± 0.1 µM for PGV-1, respectively, while not toxic against fibroblast cells. Both compounds instigated G2/M arrest and efficiently induced cell senescence and apoptosis. Moreover, these analogs selectively elevated oxidative stress in colon cancer cells without inducing noticeable changes in fibroblasts. In conclusion, PGV-1 and CCA-1.1 suppressed colorectal tumor formation and induced mitotic arrest.

Clinicopathological study of sarcomatoid renal cell carcinoma in animals in East Java, Indonesia, from 2017 to 2022.

Background: Renal cell carcinoma (RCC) is common cancer derived from the renal epithelium. One of the rarest cases of RCC is sarcomatoid RCC (sRCC). The occurrence of sRCC in animals is not clearly demonstrated. Aim: This study aimed to observe the clinicopathological characteristics of sRCC in animals from East Java, Indonesia, from 2017 to 2022. Methods: This study used patients who were histopathologically diagnosed with sRCC in our laboratory from 2017 to 2022. The data on the clinical characteristics of animals, hematology, serology, histopathology, and immunohistochemistry (IHC) were retrieved and tabulated. The data were qualitatively and quantitatively analyzed using a simple descriptive method and Statistical Package for the Social Sciences version 26, respectively. Results: Fourteen cases of sRCC in animals have been identified in this study. It was found in rodents, dogs, and cats. sRCC predominantly occurred in rodents (57.14%) without specific clinical signs. The common histopathological findings of sRCC were epithelial renal cells transition into elongated atypical spindle cells. In addition, other histopathological patterns of a renal epithelial cell such as clear cell, tubule-cystic, and papillary also have been found. IHC by using antibodies demonstrates that PAX8 is expressed on sRCC tissue samples 92.85% (13/14 samples). Hence, PAX8 could be used as a supporting method for establishing the diagnosis of sRCC in animals. Hematology and serological tests did not correlate to the type of sRCC either pure sRCC or dedifferentiated sRCC. sRCC results in hypercreatinemia in rodents and dogs. Conclusion: This study shows that the incidence of sRCC in animals is rare. Animals with sRCC did not show any specific clinical signs. The histopathological finding is quite difficult to be differentiated from the other RCC. PAX8 expression on renal tissue samples is useful in supporting the diagnosis of sRCC in animals.

A Review of the GSTM1 Null Genotype Modifies the Association between Air Pollutant Exposure and Health Problems.

Air pollution is one of the significant environmental risks known as the cause of premature deaths. It has deleterious effects on human health, including deteriorating respiratory, cardiovascular, nervous, and endocrine functions. Exposure to air pollution stimulates reactive oxygen species (ROS) production in the body, which can further cause oxidative stress. Antioxidant enzymes, such as glutathione S-transferase mu 1 (GSTM1), are essential to prevent oxidative stress development by neutralizing excess oxidants. When the antioxidant enzyme function is lacking, ROS can accumulate and, thus, cause oxidative stress. Genetic variation studies from different countries show that GSTM1 null genotype dominates the GSTM1 genotype in the population. However, the impact of the GSTM1 null genotype in modifying the association between air pollution and health problem is not yet clear. This study will elaborate on GSTM1's null genotype role in modifying the relationship between air pollution and health problems.

Anti-rheumatoid activity of a hexane-insoluble fraction from Plantago major in female Wistar rats induced by Complete Freund's Adjuvant.

Background and aim: Plantago major has long been used for medical purposes in Indonesia. However, reports on the anti-arthritic activities of P. major are limited. Experimental procedure: The anti-arthritic properties of an n-hexane-insoluble fraction of dichloromethane extracts of P. major (IPM) were evaluated using Complete Freund's Adjuvant (CFA)-induced arthritis induced in female Wistar rat by CFA. Diclofenac was used as a positive control. The volume of paw oedema, white blood cell count, lymphocytes, neutrophils, expression of TNF-α and Interleukin-6 and the histopathological features of the joint tissues were assessed to characterise IPM activity. Results: The IPM extract at doses of 280 and 420 mg/kg BW and diclofenac inhibited paw oedema by 15.70 %, 15.94 % and 19.71 % respectively. IPM also reduced the incidence of arthritis and arthritic index. Unlike untreated rats, animals treated with IPM showed a significant decrease in the number of neutrophils and decreased expression of TNF-α and Interleukin-6. Histopathological examination showed a reduction in the number of inflammatory cells and hyperplasia of the synovium after IPM treatment. Conclusion: This study showed that P. major displays anti-rheumatoid arthritis activity.

Chemopreventive Effect of Dietary Maranta arundinacea L. Against DMBA-Induced Mammary Cancer in Sprague Dawley Rats Through the Regulation of Autophagy Expression.

BACKGROUND: Breast cancer prevention still needs to be improved. Calorie restriction is thought to prevent breast cancer through the induction of autophagy. Maranta arundinacea L. (MA) has the potential for calorie restriction because it contains high fiber. This research aimed to observe the effect of dietary MA against dimethylbenz(a)anthracene (DMBA)-induced mammary cancer in Sprague Dawley rats related to autophagy. METHODS: Twenty-five Sprague Dawley rats were randomly divided into five groups: 1) control group without DMBA-induced with a standard diet, 2) 20 mg/kg BW of DMBA two times a week for five weeks with a standard diet, 3) DMBA and diet modification with 30% of MA, 4) DMBA and diet modification with 45% of MA, and 5) DMBA and diet modification with 60% of MA. Examination of the nodule was conducted once every week for 22 weeks. Breast tissue/tumor examination underwent histology examination with hematoxylin-eosin. Examinations of immunohistochemical staining against Beclin1, LC3B, and SQSTM1 were conducted to reveal autophagy. The difference of autophagy protein expression was analyzed using One way ANOVA with 95% confidence level and significance set as p<0.05. RESULTS: Cancer was detected in four rats of DMBA standard diet, two rats of 30% MA, one rat of 45% MA. No cancer was detected in the rats of control and rats with 60% of MA group. The Beclin1 expressions showed that the 60% of MA group had the highest score (2.5±0.52) followed by the 45% of MA group (1.87±0.49), control group (1.77±0.11), 30% of MA group (1.28±0.75), and DMBA with standard diet had the lowest score (1.28±0.91). The difference of Beclin1 expressions was statistically significant (p-value=0.03). However, the difference of the LC3B expressions (p-value=0.11) and SQSTM1 expressions (p-value=0.225) were not statistically significant. CONCLUSION: Dietary modifications with MA potentially prevent breast cancer and induce initiation of autophagy.

In vivo anti-inflammatory activities of Plantago major extract and fractions and analysis of their phytochemical components using a high-resolution mass spectrometry.

Background and purpose: Plantago major has been applied as a herbal remedy for centuries. However, studies on anti-inflammatory activities and their chemical ingredients are limited. The objective of this study was to investigate the anti-inflammatory properties of P. major in three animal models and its phytochemical contents. Experimental approach: Dichloromethane extract (DCM) of P. major was fractionated with n-hexane to yield the soluble (SHF) and insoluble (IHF) fractions. The anti-inflammatory activities of DCM, SHF, and IHF were evaluated using rat's paw edema induced by carrageenan, thioglycolate-induced leukocyte emigration in the mice, and rheumatoid arthritis (RA) induced by complete Freund's adjuvants in rats. The chemical constituents were analyzed using a high-resolution mass spectrometer (HRMS). Findings / Results: The DCM, SHF, and IHF inhibited paw edema in the rats and reduced the leukocyte migration in the mice. At dose 560 mg/kg, the percentage of inhibitory was 47.33%, 55.51%, and 46.61% for the DCM, IHF, and SHF, respectively. In the RA animal model, IHF at 280 and 560 mg/kg reduced osteoclast formation and COX-2 expression compared to diclofenac. Some compounds namely oleic acid, linoleic acid, palmitic acid, and oleamide identified in the DCM, IHF, and SHF may be responsible for these activities. Conclusion and implications: This study showed that P. major has several in-vivo anti-inflammatory activities.

Comparative safety and efficacy of two bivalent vaccines containing Newcastle disease LaSota and avian influenza H9N2 Sidrap isolate formulated with different oil adjuvants.

BACKGROUND AND AIM: Newcastle disease (ND) and avian influenza (AI) are two devastating diseases of poultry, which cause great economic losses to the poultry industry and disrupt food security in our country. The use of ND-AI inactive bivalent vaccine is very effective and economical to prevent and control ND and AI disease. Bivalent ND LaSota-AI H9N2 vaccine is not yet available in Indonesia. The inactivated vaccines used in poultry industry often require oil adjuvant to elicit a sufficient immune response. This study aimed to develop the bivalent inactive vaccines containing ND LaSota and AI H9N2 Sidrap isolate which are local isolates as poultry vaccine candidates, and formulated with two different commercial adjuvants, then compared. MATERIALS AND METHODS: Two vaccines bivalent were prepared by emulsifying inactivated Newcastle disease virus (LaSota strain) and AI H9N2 Sidrap isolate viruses with Marcol white mineral oil and Montanide ISA70 adjuvants. Both of bivalent vaccines were tested for safety (physical and histopathological at the injection site) and efficacy in specific-pathogen-free chickens. Parameters used for the evaluation of the efficacy were immunogenicity by hemagglutination inhibition and protection percentage. RESULTS: Both bivalent vaccines are safe to use. Post-vaccination (PV) immune response was observed using a hemagglutination inhibition test at 2, 3, 4, 5, 6, 7, and 8 weeks of PV. The bivalent vaccine B gives a better immune response to ND at 2, 3, and 4 weeks of PV (p<0.05) compared to the bivalent vaccine A, but in 5, 6, 7, and 8 weeks, the PV does not show differences in the immune response. The immune response to AI H9N2 showed differences at weeks 2 and 3 PV (p<0.05) with the bivalent vaccine B indicated higher immunity. A single immunization with both bivalent vaccines induces 100% protection in chickens that have been vaccinated against the deadly challenge with the virulent ND virus. CONCLUSION: Both of bivalent vaccines are safe to use and provide good efficacy against virulent ND viruses, but bivalent vaccine B (with Montanide ISA70 adjuvant) shows better immune response than bivalent vaccine A (Marcol white mineral oil adjuvant).

Dermatophytosis in companion animals: A review.

Dermatophytosis, a zoonotic disease, is caused by fungi of three main genera, namely, Micropsorum, Trichophyton, and Epidermophyton. Specific lesions of dermatophyte infections are localized in the face, legs, and/or tail. Skin lesions in infected animals demonstrate localized alopecia, erythema, and crust, which are more commonly known as ringworm. Factors that affect dermatophytosis include the dermatophyte species; virulence factors of the agent; and the immune status, age, and sex of the host. High levels of cortisol and pro-inflammatory cytokines have also been reported to play an important role in dermatophyte infection. This review aims to explore and understand factors that affect dermatophyte infection with an emphasis on the prevalence, clinical signs, pathogenesis, immune response, and the roles of cortisol and cytokines in companion animals infected by a dermatophyte.

The Effect of (E)-1-(4'-aminophenyl)-3-phenylprop-2-en-1-one on MicroRNA-18a, Dicer1, and MMP-9 Expressions against DMBA-Induced Breast Cancer.

BACKGROUND: Most of breast cancer patients are estrogen receptor alpha-positive and have high resistance and side effect of chemotherapeutic drug. Therefore, discovering an effective anticancer agent is needed. This research explored the effect of (E)-1-(4'-aminophenyl)-3-phenylprop-2-en-1-one (APE) on miR-18a, Dicer1, and MMP-9 expressions. METHODS: Twenty four female Sprague-Dawley rats were invetigated in this study. The rats were divided into 6 groups of 4. G1 was considered as normal rat. G2, G3, T1, T2, and T3 were given DMBA 20 mg/kgBW twice a week for 5 weeks to induce mammary cancer. After being affiliated with cancer, G2 was given vehicle and G3 was treated with tamoxifen. T1, T2, and T3 were treated with APE intraperitoneally everyday for 21 days at doses of 5, 15, and 45 mg/kgBW/day, respectively. Blood plasma was collected to measure miR-18a expression using qRT-PCR. Mammary tissues were also collected to determine Dicer1 and MMP-9 expressions by using  immunohistochemistry. RESULTS: The results showed significant down-regulation of miR-18a relative expression and up-regulation of Dicer1 expression in G3 and T1 compared to G2 (P<0.05). MMP-9 expression has significant decrease in T1 compared to G2 (P<0.05). CONCLUSION: APE can decrease miR-18a and MMP-9 expressions and increase Dicer1 expression in rat mammary cancer. Therefore, this compound could be a candidate of novel anticancer.

Clinical and pathological features of aspergillosis due to Aspergillus fumigatus in broilers.

BACKGROUND AND AIM: Aspergillus fumigatus is a ubiquitous pathogen causing aspergillosis in poultry. This research aimed to evaluate the clinical and pathological features of aspergillosis infection in broilers. MATERIALS AND METHODS: A. fumigatus infection was induced experimentally by intra-air sac inoculation of a 1.7×108 spore suspension into broilers. Infected and non-infected birds were closely observed for the development of clinical signs of infection twice daily. Pathological samples were collected 5, 14, and 30 days post-infection (dpi) and examined by hematoxylin-eosin staining. RESULTS: A total of 160 birds were included in this study. Clinical signs emerged at 3 dpi and became consistent at 5 dpi. A considerable decrease in severity and number of birds showing infection symptoms followed. The clinical signs of aspergillosis included anorexia (n=40; 50%), lethargy (n=32; 40%), dyspnea (n=38; 48%), and gasping (n=29; 36%). Macroscopic changes in the air sacs at 3 dpi included the development of minor lesions showing cloudiness, slight membrane thickening, and local exudates. Histopathological examination of the air sacs collected at 3 dpi indicated local inflammation surrounded by hyphae and spores. At 5 dpi, infected birds developed nodules, necrosis, and parenchymal consolidation of the lungs. Pulmonary changes, such as bronchopneumonia, spores, septate hyphae, and mild granulomatous inflammation, were also observed. At 14 dpi, multiple caseous nodules and plaques were found in the air sacs; plaque and necrosis in large areas of the lungs and severe multifocal granulomatous inflammation were noted. CONCLUSION: The clinical symptoms of aspergillosis emerged at 3 dpi and gradually decreased beginning at 7 dpi. Similar pathological changes were observed in the air sacs and lungs. The results of this work provide additional information on the pathogenesis of aspergillosis.

Efficacy of tetravalent coryza vaccine against the challenge of Avibacterium paragallinarum serovars A and B isolates from Indonesia in chickens.

AIM: Infectious coryza is caused by Avibacterium paragallinarum. In Indonesia, this infection results in a 10%-40% decrease in egg production by laying hens. This study was conducted to determine the effectiveness of tetravalent coryza vaccine contained A. paragallinarum bacterin serovars A, B, C2, and C3; strain A-221, B-Spross, C2-Modesto, and C-3-Akko in layers based on antibody titer and clinical signs using a post-challenge test. MATERIALS AND METHODS: Forty four-week-old Lohmanns strain chickens were used in this study. Forty chickens were divided into four groups for serological and challenge test: Group 1 (unvaccinated and challenged by A. paragallinarum serovar A), Group 2 (unvaccinated and challenged by A. paragallinarum serovar B), Group 3 (vaccinated and challenged by A. paragallinarum serovar A), and Group 4 (vaccinated and challenged by A. paragallinarum serovar B). Vaccination was done using the tetravalent vaccine in oil-emulsion adjuvant contained A. paragallinarum bacterin serovars A, B, C2, and C3; strain A-221, B-Spross, C2-Modesto, and C-3-Akko. Vaccination was performed at day 1 and booster was done at day 14. Blood serum was collected on days 0, 14, and 28 for the hemagglutination-hemagglutination inhibition (HI) test. The challenge test was given at day 29 through intranasal administration using A. paragallinarum serovars A-L2447 and B-L1710 approximately 6×108 CFU/mL. Clinical signs were observed for 14 days post-infection. At the end of the study, chickens were euthanized, and pathological features of the infraorbital sinus, facial skin, and trachea were recorded. RESULTS: Data analysis of antibody titers and pathological changes was performed descriptively, while clinical symptom scores were analyzed non-parametrically with the Mann-Whitney U-test using SPSS version 21. At days 14 and 28 post-vaccination, the antibody titer in Group 3 was 5 HI and 20 HI, respectively. However, the antibody titers in Group 4 at 28 days post-vaccination were 0 HI. Clinical observations, the vaccinated groups that were challenged with A. paragallinarum serovars A and B showed clinical symptoms on days 4 and 6 post-infection, namely mild unilateral facial edema and severe bilateral facial edema, respectively. Clinical signs in Groups 3 and 4 were less severe than in Groups 1 and 2 (p<0.05). Pathological examination findings supported clinical observations and serological testing. CONCLUSION: Tetravalent coryza vaccine in chickens has efficacy to protect against the challenge test of A. paragallinarum serovars A and B isolated from Indonesia.

Combinations of herbs and probiotics as an alternative growth promoter: An in vitro study.

BACKGROUND AND AIM: Antibiotic growth promoters (AGPs) are added to animal feed to stimulate growth and increase livestock productivity. However, the regular use of antibiotics in animal diets has a considerable contribution to the occurrence of antibiotic resistance in livestock and humans. This study aimed to investigate the feasibility of red ginger (Zingiber officinale var. Rubrum), turmeric (Curcuma domestica), and wild ginger (Curcuma xanthorrhiza), Lactobacillus acidophilus, and Lactobacillus brevis as an alternative to AGPs. MATERIALS AND METHODS: The antibacterial activities and probiotic stimulatory effects of herbs were screened through the disk diffusion method and optical densitometry. The inhibitory ability of probiotics against pathogens was also tested through the disk diffusion method. The adhesion ability of probiotics was tested by mixing the optimal herbal combinations with broiler intestinal epithelial cells (105 cells/ml). The cells were then subjected to Gram staining, and the number of adherent bacteria was calculated. RESULTS: The test results showed that 3.13% ethanolic wild ginger extract had the highest inhibitory activity against Salmonella Enteritidis, followed by ethanolic red ginger extract and aqueous wild ginger extract at the same concentration. The three extracts also supported the growth of L. acidophilus and L. brevis. Further tests showed that the combination of 3.13% ethanolic red ginger extract had the highest inhibitory activity against S. Enteritidis, followed by ethanolic and aqueous wild ginger extract at the same concentration. The three extracts also supported the growth of L. acidophilus and L. brevis. Further tests showed that the combination of 3.13% ethanolic red ginger extract and 3.13% aqueous wild ginger extract had the best inhibitory effect on the growth of S. Enteritidis. The stimulatory effect of the combinations of herbal extract on the growth of L. acidophilus (0.18±0.00) and L. brevis (0.21±0.01) was better than those of individual extract, positive controls, and the glucose control. L. acidophilus and L. brevis had a weak inhibitory effect on the growth of S. Enteritidis (<6 mm). The adhesion ability of L. acidophilus (420.00±28.21) and L. brevis (259.33±24.03) was stronger than that of S. Enteritidis (202.00±14.00) under treatment with combined extracts. CONCLUSION: The tested combinations of herbs and probiotics can adhere to the intestinal tract. Given this characteristic, herb and probiotic combinations may be developed as an alternative to conventional AGPs.

CCL2 and IL18 expressions may associate with the anti-proliferative effect of noncontact electro capacitive cancer therapy in vivo.

Background: Noncontact Electro Capacitive Cancer Therapy (ECCT) is a novel treatment modality in cancer. Chemokine (C-C motif) ligand 2 (CCL2) has a major role in the outgrowth of metastatic breast cancer. Interleukin 18 (IL18) plays a role in macrophage alteration, which leads to excessive angiogenesis. This study aims to elaborate on the association of CCL2, IL18, IL23α, and TNF-α (tumor necrosis factor-alpha) expression with the anti-proliferative effect of ECCT in rat breast tumor tissue.   Methods: Low intensity (18 Vpp) and intermediate frequency (150 kHz) alternating current-electric field (AC-EF) between two capacitive electrodes were exposed as external EF to a rat cage. Twenty-four rats were divided into four groups of six replicates. Breast tumor tissues were collected from 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rats. Two groups were non DMBA-induced rats without ECCT exposure (NINT) and with (NIT). The other two groups were DMBA-induced rats without ECCT exposure (INT) and with (IT). Mammary glands and breast tumor tissues were collected from each group and preserved. Hematoxylin-eosin and immunohistochemistry staining were performed on paraffin sections of tissues using anti-PCNA, anti-ErbB2, anti-Caspase3, and anti-CD68. CCL2, IL18, IL23α, and TNF-α mRNA relative expressions were analyzed using qRT-PCR. Results: ECCT exposure may cause the reduction of PCNA protein expression as well as ErbB2 on breast tumor tissues, but it causes the increase of Caspase3 and macrophage CD68 protein. In rat breast tumor tissues of IT groups, the mRNA expression of CCL2 and IL18 are significantly down-regulated, in contrast with the up-regulated expression of these cytokines in tumor tissues of the INT group. IL23α and TNF- α expression remained similar in both groups. Conclusion: CCL2 and IL18 expressions have an association with the inhibition of breast tumor cell proliferation affected by ECCT exposure.

Chemopreventive Effects of Edible Canna (Canna edulis Kerr.) Against Colorectal Carcinogenesis: Effects on Expression of Adenomatous Polyposis Coli and Inducible Nitric Oxide Synthase in Rat Inflammatory Model

Objective: Dietary high fibre and calcium intake has been suggested to reduce colorectal cancer risk. However, there is limited information available regarding the potential of edible canna (Ganyong), with high dietary fibre and calcium content, to act as a preventive agent for colorectal cancer. This experimental study was conducted to investigate the preventive effect of Ganyong in reducing colorectal carcinogenesis with attention to effects on adenomatous polyposis coli (APC) and inducible nitric oxide synthase (iNOS) expression. Methods: Thirty male Wistar rats were divided into 5 equal groups; a normal control group without azoxymethane/dextran sodium sulphate (AOM/DSS) induction and Ganyong, a 'cancer' control group with AOM/DSS induction only, and three treatment groups with AOM/DSS induction and different percentages (5%, 10% and 20%) of Ganyong. Paraffin-embedded sections of rat colon tissue were analysed by haematoxylin-eosin and immunohistochemical staining against antibodies against APC and iNOS. Variation in rates of APC and iNOS expression were analyzed using the Kruskal-Wallis test followed by the Dunn's test (SPSS statistic version 24). P<0.05 was considered statistically significant. Results: AOM/DSS induction increased the expression of APC (p=0.013) and iNOS (p=0.013) compared to the normal control group. APC expression in the treated groups was lower than in the 'cancer' control group (p=0.049), especially in the 10% Ganyong group (p=0.02). In contrast, there was no significant variation among the treated groups regarding iNOS expression. Histopathological features of the colon supported the data for APC and iNOS expression. Conclusion: This study indicated potential chemopreventive effects of Ganyong reducing expression of factors contributing to colorectal carcinogenesis.

Protection by Ethanolic Extract from Ulva lactuca L. against Acute Myocardial Infarction: Antioxidant and Antiapoptotic Activities.

BACKGROUND: Reactive oxygen species (ROS) play a major role in myocardial damage during acute myocardial infarction (AMI). This study aimed to determine the antioxidant and antiapoptotic activities of an ethanolic extract from Ulva lactuca L. (EEUL) against AMI. METHODS: Thirty-six male Wistar rats were divided into six groups: one control group and five treatment groups. Treatment group II was given 85 mg/kg body weight (BW) of isoproterenol (ISO). Group III, IV and V were given ISO and EEUL at 250, 500 and 750 mg/kg BW, respectively. Group VI were given 10 mg/kg BW of ISO and melatonin. EEUL and melatonin were orally administered for 28 days. ISO was injected subcutaneously on day 29 and 30 to chemically induce AMI. On day 31, blood was collected for antioxidant assay and heart tissues were collected for histological examination. RESULTS: The activity of catalase (CAT), an endogenous antioxidant, in the EEUL-treatment groups was significantly increased compared to the ISO-treatment group (P < 0.001). The EEUL-treatment groups showed significantly decreased expression of caspase-3 (P < 0.001) and better myocardial tissue morphology. CONCLUSION: EEUL possibly protects against AMI because of its antioxidant and antiapoptotic properties.

Photoimmune protective effect of the phytoestrogenic isoflavonoid equol is partially due to its antioxidant activities.

Topical application of lotions containing the phytoestrogenic isoflavonoid equol have been reported to protect mice against UV radiation-induced inflammation, immune suppression and photocarcinogenesis. The photoimmune protective property was shown to depend on equol's activation of oestrogen receptor signalling in the skin. However, isoflavones are also recognised for their antioxidant properties in biological systems. As endogenous cutaneous antioxidant enzymes including the inducible stress protein haem oxygenase (HO)-1, have photoprotective efficacy, this study in the Skh:hr-1 hairless mouse seeks evidence for an antioxidant role for equol in contributing to its photoimmune protection. Oxidative stress has been measured as UVA-induced lipid peroxidation in the mouse skin, and was dose-dependently inhibited by topical equol. Inhibition of the UVA (320-400 nm)-inducible HO activity significantly reduced the level of equol protection against lipid peroxidation, thereby attributing a component of equol's lipid protection capacity to this stress enzyme. It was consistent that topical equol enhanced the level of HO induction by UVA irradiation in both skin and liver. Subsequently, the dose-dependent protection by topical equol lotions against solar simulated UV radiation induced immunosuppression, measured by the contact hypersensitivity reaction, was found also to be partially reduced by the inhibition of HO activity. Therefore, in addition to the activation by equol of oestrogenic signalling pathways for photoprotection, this isoflavonoid also provides UV-protective antioxidant effects that depend partially on HO-1 induction.

Topical isoflavonoids reduce experimental cutaneous inflammation in mice.

The antioxidant and anti-proliferative biological effects of isoflavonoids are relevant properties to counteract the characteristics of many cutaneous diseases. This study uses ultraviolet (UV)B irradiation to induce inflammation in the mouse skin, as a model for some symptoms of cutaneous inflammatory and hyperproliferative diseases such as psoriasis in humans, with the objective of testing two topically applied isoflavonoid compounds for therapeutic properties. UVB exposure resulted in the overexpression of the cytokines, tumour necrosis factor (TNF)-α, interleukin (IL)-6 and the adhesion molecule P-cadherin. Infiltration into the dermal compartment of mast cell populations was also induced. These factors are also overexpressed in psoriatic skin. The effect of topical applications of two isoflavonoids, equol and a synthetic analogue NV-38, was tested. Both isoflavonoids dose dependently inhibited the UVB induction of cutaneous TNF-α mRNA and protein, a cytokine critical for the initiation of psoriatic inflammation. Expression of IL-6 mRNA and protein was also decreased, and the number of infiltrating mast cells into the dermis was reduced by both isoflavonoids. Furthermore, the upregulated mRNA and protein levels of P-cadherin, a marker characteristic of cutaneous hyperproliferation, were also normalized by both isoflavonoids. These results suggest that this class of compounds has the potential for useful, innocuous anti-inflammatory therapy from topical application in human cutaneous diseases.

Estrogen receptor signaling protects against immune suppression by UV radiation exposure.

The phytoestrogenic isoflavonoid equol is known to protect against solar-simulated UV radiation-induced inflammation, immunosuppression, and skin carcinogenesis. The mechanism may involve antioxidant actions, because equol not only is a radical scavenger but also enhances the induction of a relevant cutaneous antioxidant, metallothionein. However, this study in female hairless mice examined whether the estrogenicity of the isoflavonoid might be responsible. Protection by topically applied equol against photoimmune suppression was found to be strongly and dose-dependently inhibited by the estrogen receptor (ER) antagonist ICI 182,780. Furthermore, ICI 182,780 alone was found to significantly exacerbate immunosuppression resulting from solar-simulated UV radiation irradiation, suggesting a natural role for the ER in photoimmune protection. In support of this role, topical application of the physiological ligand 17-beta-estradiol also provided dose-dependent photoimmune protection, inhibitable by ICI 182,780, that was attributed largely to the inactivation of the downstream actions of cis-urocanic acid, an important endogenous immunosuppressive photoproduct. Thus, a hitherto unrecognized function of the ER as a normal photoprotective immune regulator in the skin was revealed. The relationship between equol and cutaneous metallothionein suggests an association of the ER with this inducible antioxidant in constraining the photoimmune-suppressed state and therefore in the prevention of the facilitation of photocarcinogenesis by this immunological defect. This role for the ER may underlie important gender-specific differences in UV-responsiveness that would reflect different needs for environmental photoprotection in males and females.

Protective effect of the isoflavone equol against DNA damage induced by ultraviolet radiation to hairless mouse skin.

Equol, an isoflavonoid metabolite produced from the dietary isoflavone daidzein by the gut microflora in mammals, has been found to protect not only against ultraviolet (UV) radiation-induced cutaneous inflammation and photoimmune suppression, but also have antiphotocarcinogenic properties in mice. Because the state of DNA damage has been correlated with suppression of the immune system and photocarcinogenesis, we have therefore examined the potential of equol to offer protection from solar-simulated UV (SSUV) radiation-induced DNA damage in hairless mice by the immunohistochemical approach using monoclonal antibody specific for cyclobutane pyrimidine dimers (CPDs; H3 antibody). Topical application of 20 microM equol lotion, which was applied both before and after SSUV significantly reduced the number of CPDs. This reduction was evident immediately after SSUV exposure, at 1 h after exposure, and at 24 h after exposure, revealing 54%, 50%, and 26% reduction in CPDs, respectively. When the same concentration was applied for 5 consecutive days after SSUV exposure, there was no significant difference in the reduction of CPDs immediately after SSUV irradiation or at 1 hour afterwards, but there were significant reductions of 23% and 42% at 24 and 48 h after SSUV exposure, respectively. Despite apparently reducing the number of CPDs post-SSUV, topically applied equol did not appear to increase the rate of dimer removal. To conclude, equol applied topically prior to SSUV irradiation offers protection against CPD formation in hairless mice, possibly by acting as a suncreen and thus inhibiting DNA photodamage.