Canine Visceral Leishmaniasis: A Histological and Immunohistochemical Study of Fibropoiesis in Chronic Interstitial Pneumonitis.

We studied some fibrotic aspects of chronic interstitial pneumonitis in the lungs of dogs infected with Leishmania infantum. The lungs of eleven naturally infected dogs, twelve experimentally infected with two distinct strains of L. infantum (BH401 and BH46), and six uninfected (controls) dogs, were analyzed by histological, parasitological, and immunohistochemical studies. Conventional histology (HE), collagen deposition (Gomori's silver staining for reticulin collagen fibers), and immunohistochemistry for myofibroblast characterization were carried out based on the cellular expression of alpha-smooth muscle actin, vimentin, cytokeratin, E-cadherin, snail antigen homologue 1 (SNAI1) (Snail), and the cytokine expression of transforming growth factor-beta (TGF-ß). Parasitological screening was carried out using conventional polymerase chain reaction (PCR) and the immunohistochemical reaction of streptavidin-peroxidase for visualizing Leishmania amastigotes. Dogs naturally infected with L. infantum and experimentally infected with L. infantum BH401 strains showed intense interstitial pneumonitis characterized by thickening of the alveolar septa as a consequence of an intense diffuse and focal (plaques) chronic exudate of mononuclear cells associated with fibrogenesis. The expression of alpha-actin, vimentin, and TGF-ß was higher in the lung interstitium of all infected dogs than in the other two groups (BH46 strain and controls). Moreover, in both the naturally and experimentally infected dog (BH401 strain) groups, the expression of Snail was moderate to intense in contrast to the other groups. Based on these immunohistochemical results, we concluded that mesenchymal cells are active in promoting changes in the extracellular matrix in the lungs of dogs naturally and experimentally infected with L. infantum, but it depends on the virulence of the parasite.

Photodynamic therapy repairs medication-related osteonecrosis of the jaw by reducing NF-kB protein in rats.

OBJECTIVE: To evaluate whether antimicrobial photodynamic therapy (aPDT) repairs bisphosphonate-related osteonecrosis of the jaw (BRONJ) modulated by the reduction of NF-kB protein in a murine model. METHODOLOGY: Male Wistar rats (N=30) were divided into the following groups (n=6/group): negative control (NC); experimental osteonecrosis (ONE); ONE + photosensitizer (PS); ONE + photobiomodulation (PBM); and ONE + aPDT. Over 8 weeks, ONE was induced by zoledronic acid 250 µg/kg injections, except in the NC group, which received sterile 0.9% saline, followed by extraction of the lower left first molar. Red light laser irradiation (wavelength ~660 nm, power 50 mW, energy of 2 J, energy dose of 66.67 J/cm2 for 40 s) was performed once a week for 4 weeks. Methylene blue 0.3% was used as PS. The animals were euthanized and examined macroscopically for the presence of exposed bone and epithelial repair and microscopically by histochemical (hematoxylin-eosin and Masson's trichrome staining) and immunohistochemical (anti-NF-kB) methods. Macroscopic and histomorphometric data were analyzed by one-way ANOVA and Tukey's post-test (p<0.05). RESULTS: Mucosal repair, viable osteocytes, and NF-kB immunostaining were observed in the NC, ONE+PS, ONE+PBM, and ONE+aPDT groups. The ONE group showed no mucosal repair, showing empty lacunae and multifocal immunostaining for NF-kB. The ONE+PBM and ONE+aPDT groups had greater deposition of extracellular matrix and less necrotic bone tissue (p<0.05). CONCLUSION: PBM and aPDT treatments for BRONJ were effective for bone and epithelial repair, in addition to reducing inflammation mediated by the decrease of NF-kB protein in the irradiated regions.

(-)-Fenchone Prevents Cysteamine-Induced Duodenal Ulcers and Accelerates Healing Promoting Re-Epithelialization of Gastric Ulcers in Rats via Antioxidant and Immunomodulatory Mechanisms.

BACKGROUND: (-)-Fenchone is a naturally occurring monoterpene found in the essential oils of Foeniculum vulgare Mill., Thuja occidentalis L., and Peumus boldus Molina. Pharmacological studies have reported its antinociceptive, antimicrobial, anti-inflammatory, antidiarrheal, and antioxidant activities. METHODS: The preventive antiulcer effects of (-)-Fenchone were assessed through oral pretreatment in cysteamine-induced duodenal lesion models. Gastric healing, the underlying mechanisms, and toxicity after repeated doses were evaluated using the acetic acid-induced gastric ulcer rat model with oral treatment administered for 14 days. RESULTS: In the cysteamine-induced duodenal ulcer model, fenchone (37.5-300 mg/kg) significantly decreased the ulcer area and prevented lesion formation. In the acetic acid-induced ulcer model, fenchone (150 mg/kg) reduced (p < 0.001) ulcerative injury. These effects were associated with increased levels of reduced glutathione (GSH), superoxide dismutase (SOD), interleukin (IL)-10, and transforming growth factor-beta (TGF-ß). Furthermore, treatment with (-)-Fenchone (150 mg/kg) significantly reduced (p < 0.001) malondialdehyde (MDA), myeloperoxidase (MPO), interleukin-1 beta (IL-1ß), tumor necrosis factor-alpha (TNF-α), and nuclear transcription factor kappa B (NF-κB). A 14-day oral toxicity investigation revealed no alterations in heart, liver, spleen, or kidney weight, nor in the biochemical and hematological parameters assessed. (-)-Fenchone protected animals from body weight loss while maintaining feed and water intake. CONCLUSION: (-)-Fenchone exhibits low toxicity, prevents duodenal ulcers, and enhances gastric healing activities. Antioxidant and immunomodulatory properties appear to be involved in its therapeutic effects.

Intermittent fasting associated with coconut oil (Cocos nucifera L.) alters gut-liver axis parameters in diet-induced obese rats.

OBJECTIVE: The aim of this article is to investigate the effect of intermittent fasting, associated or not with coconut oil intake, on the gut-liver axis of obese rats. METHODS: A total of 50 rats were divided into five groups: control, obese, obese with intermittent fasting, obese with intermittent fasting plus coconut oil, and obese with caloric restriction. The rats were induced to obesity with a high-sugar diet for 17 wk. The respective interventions were carried out in the last 4 wk. RESULTS: The groups with intermittent fasting protocols had reduced total cholesterol (on average 54.31%), low-density lipoprotein (on average 53.39%), and triacylglycerols (on average 23.94%) versus the obese group; and the obese with intermittent fasting plus coconut oil group had the highest high-density lipoprotein compared with all groups. The obese with intermittent fasting plus coconut oil and obese with caloric restriction groups had lower metabolic load compared with the other groups. The obese group had high citric and succinic acid concentrations, which affected the hepatic tricarboxylic acid cycle, while all the interventions had reduced concentrations of these acids. No histologic changes were observed in the intestine or liver of the groups. CONCLUSION: Intermittent fasting, especially when associated with coconut oil, had effects comparable with caloric restriction in modulating the parameters of the gut-liver axis.

Identification of phlebotomine sand fly (Diptera: Psychodidae) in Atlantic forest fragments and their dispersal to urban area.

The geographical distribution of sand flies in Brazil has been the subject of some studies, yet there is no information about the phlebotomine fauna in João Pessoa, State of Paraíba, Brazil. The aim of this work is to evaluate the occurrence and distribution of sand flies in the Atlantic forest fragments and to evaluate a possible dispersion in 06 nearby districts. Light traps were used during three consecutive nights, supplemented by an aspirator during the dry period and rainy season. A total of 222 sand flies were found, 143 (130 males and 13 females) in the Atlantic forest, and 79 in urban areas. During the entire dry season, three species of phlebotomine sand flies were recorded in 11 forest fragments, Lutzomyia longipalpis, Lu. migonei and Lu. whitmani. During the rainy season, only Lu. longipalpis was found. This was the only species identified in the studied neighborhoods during both seasons. The differences in diversity of sand flies encountered between natural habitats and urban areas may thus be correlated mostly with adaptations to particular habitats and availability of food. One species (Lu. longipalpis) appears to be rapidly adapting to urban areas because of deforestation.

Obesity influences the development of bisphosphonate-induced osteonecrosis in Wistar rats.

Medication-related osteonecrosis of the jaw (MRONJ) is characterized by bone exposure for more than eight weeks in patients who have used or been treated with antiresorptive or antiangiogenic drugs, without a history of radiation therapy or metastatic diseases in the jaws. Obesity is associated with changes in periodontal tissues and oral microbiota that are linked to bone alterations. This study aimed to analyze the influence of obesity on the development of bisphosphonate-induced osteonecrosis. The experiment randomly and simply divided 24 male Wistar rats (Rattus norvegicus) into four groups: healthy, with osteonecrosis, obese, and obese with osteonecrosis (n=6 per group). Osteonecrosis was induced through weekly intraperitoneal injection for eight weeks at a dose of 250 µg/kg of zoledronic acid in a 4 mg/5 mL solution, combined with trauma (exodontia). Obesity was induced through a high glycaemic index diet. Each group was qualitatively and quantitatively evaluated regarding the development of models and pathological anatomy of the lesions. The results were expressed in mean percentage and standard deviation and statistically analyzed using one-way analysis of variance (ANOVA) followed by Tukey's post-hoc test, with a significance level of 5% (p<0.05) to establish differences found between the groups. Animals in the osteonecrosis group and the obese with osteonecrosis group presented larger necrosis areas (averages: 172.83±18,19 µm2 and 290.33±15,77 µm2, respectively) (p<0,0001). Bone sequestration, hepatic steatosis, and increased adipocyte size were observed in the obese group (average: 97.75±1.91 µm2) and in the obese with osteonecrosis group (average: 98.41±1.56 µm2), indicating greater tissue damage in these groups (p<0,0001). All parameters analyzed (through histological, morphometric, and murinometric analyses) increased for the obese and obese with osteonecrosis groups, suggesting a possible influence of obesity on the results. However, further studies are needed to confirm the role of obesity in the possible exacerbation of osteonecrosis and understand the underlying mechanisms.

Intermittent fasting associated with aerobic exercise improves oxidative parameters and causes muscle damage without compromising the performance of Wistar rats.

OBJECTIVES: The aim of this study was to` investigate the effects of intermittent fasting (IF) and the possible association with aerobic exercise on performance, oxidative, biochemical, and somatic parameters of Wistar rats. METHODS: Forty rats were randomized into the following groups: sedentary (SC) and trained (TC) controls, sedentary intermittent fasting (SIF), and trained intermittent fasting (TIF). The rats were subjected to IF for 15 h every day and aerobic exercise lasting 30 min, five times a week, at a speed of 15 m/min for 4 wk. Performance tests were performed at the beginning and end of the protocol. Glucose and insulin tolerance, somatic parameters, lipidogram, leptin, insulin, malondialdehyde, antioxidant capacity, C-reactive protein, alpha acid glycoprotein, creatine kinase, lactate dehydrogenase, and muscle histology were analyzed. RESULTS: The trained groups had similar performance and significantly improved performance at the end of the experiment. TIF showed lower body weight (-16 g), lean mass (22.49%), homeostatic model assessment for insulin resistance (29%), and lactate dehydrogenase (48%), and higher malondialdehyde (53%) and antioxidant capacity (75%) than the TC group. The SIF and TIF groups showed a fiber area reduction and positivity marking for tumor necrosis factor-α in the muscles. CONCLUSION: Although IF associated with aerobic exercise improved antioxidant capacity caused damage to muscle fibers and lean mass loss, it did not change the performance of the rats.

Mix of almond baru (Dipteryx alata Vog.) and goat whey modulated intestinal microbiota, improved memory and induced anxiolytic like behavior in aged rats.

The objective was to evaluate the effects of the consumption of a mix of baru almond and goat whey on memory performance and anxiety parameters related to the intestinal health of rats treated during aging. The animals were divided into three groups and treated by gavage for 10 weeks (n = 10/each group): Control (CT) - distilled water; Baru almond (BA) - 2000 mg of baru/kg of body weight; and Baru + Whey (BW) - 2000 mg of baru + 2000 mg of goat milk whey/kg of body weight. Anxiety behavior, memory, brain fatty acid profile and fecal microbiota were measured. BA and BW realized less grooming, spent more time in the central area of the open field and the open arms, and realized more head dipping in the elevated plus maze. A higher rate of exploration of the new object in the short and long-term memory was observed in BA and BW. There was an increase in the deposition of MUFAs and PUFAs and oleic acid in the brain of BA and BW. Regarding spatial memory, BA and BW performed better, with an emphasis on BW. There was a beneficial modulation of the fecal microbiota with a reduction of the pathogenic genus Clostridia_UFC-014 in BA and BW and an increase in the abundance of metabolic pathways of interest in the brain-gut axis. Thus, consumption of the mix is efficient in beneficially altering the intestinal microbiota, improving memory and anxiolytic-like behavior in rats during aging.

Extra virgin coconut oil (Cocos nucifera L.) intake shows neurobehavioural and intestinal health effects in obesity-induced rats.

The present study aimed to evaluate the effect of E-VCO on the neurobehaviour and intestinal health parameters of obesity-induced rats, focusing on food consumption, body composition, bacterial and faecal organic acids and histological analyses in the hippocampus and colon. A total of 32 male Wistar rats were randomized into healthy (HG, n = 16) and obese groups (OG, n = 16), which consumed a control or cafeteria diet for eight weeks, respectively. After this period, they were subdivided into four groups: healthy (HG, n = 8); healthy treated with E-VCO (HGCO, n = 8); obese (OG, n = 8); obese treated with E-VCO (OGCO, n = 8), continuing for another eight weeks with their respective diets. The treated groups received 3000 mg kg-1 of E-VCO and control groups received water via gavage. Food preference, body weight gain, body composition, anxiety- and depression-like behaviour were evaluated. Bacteria and organic acids were evaluated in faeces, and histological analyses of the hippocampus and M1 and M2 macrophages in the colon were performed. E-VCO reduced energy intake (16.68%) and body weight gain (16%), although it did not reduce the fat mass of obese rats. E-VCO showed an antidepressant effect, increased lactic acid bacteria counts and modulated organic acids in obese rats. Furthermore, E-VCO protected the hippocampus from neuronal degeneration caused by the obesogenic diet, decreased the M1 macrophage and increased the M2 macrophage population in the gut. The results suggest neurobehavioural modulation and improved gut health by E-VCO, with promising effects against obesity-related comorbidities.

Maternal high-fat diet alters the neurobehavioral, biochemical and inflammatory parameters of their adult female rat offspring.

BACKGROUND: Lipid metabolism dysregulations have been associated with depressive and anxious behaviors which can affect pregnant and lactating individuals, with indications that such changes extend to the offspring. Therefore, the aim of this study was to evaluate the effect of a maternal high-fat diet on the neurobehavioral, biochemical and inflammatory parameters of their adult female offspring. METHODS: Wistar rats ± 90 days old were mated. The dams were allocated to consume a control (CTL) or high-fat (HFD) diet during pregnancy and lactation. After weaning, the female offspring from the CTL (N = 10) and HFD (N = 10) groups received standard chow. The offspring behavioral tests were started at 120 days old. Then, the somatic measures were evaluated followed by euthanasia, histological and biochemical analyses. RESULTS: The HFD group had less ambulation and longer immobility time in the open field test compared to the CTL. The HFD group had lower HDL (48.4%) and a higher adiposity (71.8%) and LDL (62.2%) than the CTL. The CTL had a higher organic acid concentration in the intestine, mainly acetic and butyric acids, however the HFD had a higher citric and acetic acid concentration in the brain and ischemic lesion in the hippocampus with a higher NF-κB concentration. CONCLUSION: The results demonstrate deleterious effects of a maternal HFD on the neurobehavioral and biochemical parameters of their offspring which may be associated with the role of organic acids and NF-κB in fetal programming.

Acerola fruit by-product alleviates lipid, glucose, and inflammatory changes in the enterohepatic axis of rats fed a high-fat diet.

Acerola (Malpighia emarginata) by-product (ABP) has various bioactive compounds with hypoglycaemic, antioxidant and anti-inflammatory activity. The ABP effects on the biochemical changes in the enterohepatic axis caused by a high-fat diet (HFD) remains unclear. This study assessed whether the ABP or fenofibrate administration for 28 days interferes in lipid, glucose, or inflammatory changes in the enterohepatic axis of rats fed HFD. ABP induced in the rats fed HFD a reduction in body weight, serum lipids, blood glucose, and liver fat accumulation; increased insulin tolerance, and faecal bile acid excretion; regulated organic acid synthesis, faecal and colonic microbial growth; reduced M1 macrophage and increased M2 macrophage infiltration in the colon and liver, respectively. The fenofibrate did not improve the lipid or glucose alterations in enterohepatic axis of rats fed HFD. ABP has functional/nutraceutical potential in treating HFD-induced metabolic disorders with beneficial effects on lipid and glucose metabolism, and reduction of inflammation.

Obesity influences the development of bisphosphonate-induced osteonecrosis in Wistar rats

Abstract Medication-related osteonecrosis of the jaw (MRONJ) is characterized by bone exposure for more than eight weeks in patients who have used or been treated with antiresorptive or antiangiogenic drugs, without a history of radiation therapy or metastatic diseases in the jaws. Obesity is associated with changes in periodontal tissues and oral microbiota that are linked to bone alterations. This study aimed to analyze the influence of obesity on the development of bisphosphonate-induced osteonecrosis. The experiment randomly and simply divided 24 male Wistar rats (Rattus norvegicus) into four groups: healthy, with osteonecrosis, obese, and obese with osteonecrosis (n=6 per group). Osteonecrosis was induced through weekly intraperitoneal injection for eight weeks at a dose of 250 µg/kg of zoledronic acid in a 4 mg/5 mL solution, combined with trauma (exodontia). Obesity was induced through a high glycaemic index diet. Each group was qualitatively and quantitatively evaluated regarding the development of models and pathological anatomy of the lesions. The results were expressed in mean percentage and standard deviation and statistically analyzed using one-way analysis of variance (ANOVA) followed by Tukey's post-hoc test, with a significance level of 5% (p<0.05) to establish differences found between the groups. Animals in the osteonecrosis group and the obese with osteonecrosis group presented larger necrosis areas (averages: 172.83±18,19 µm2 and 290.33±15,77 µm2, respectively) (p<0,0001). Bone sequestration, hepatic steatosis, and increased adipocyte size were observed in the obese group (average: 97.75±1.91 µm2) and in the obese with osteonecrosis group (average: 98.41±1.56 µm2), indicating greater tissue damage in these groups (p<0,0001). All parameters analyzed (through histological, morphometric, and murinometric analyses) increased for the obese and obese with osteonecrosis groups, suggesting a possible influence of obesity on the results. However, further studies are needed to confirm the role of obesity in the possible exacerbation of osteonecrosis and understand the underlying mechanisms.

Milonine attenuates the lipopolysaccharide-induced acute lung injury in mice by modulating the Akt/NF-κB signaling pathways.

Acute lung injury is an inflammation that triggers acute respiratory distress syndrome with perialveolar neutrophil infiltration, alveolar-capillary barrier damage, and lung edema. Activation of the toll-like receptor 4 complex (TLR4/MD2) and its downstream signaling pathways are responsible for the cytokine storm and cause alveolar damage. Due to the complexity of this pulmonary inflammation, a defined pharmacotherapy has not been established. Thus, this study evaluated the anti-inflammatory potential of milonine, an alkaloid of Cissampelos sympodialis Eichl, in an experimental model of lung inflammation. BALB/c mice were lipopolysaccharide-challenged and treated with milonine at 2.0 mg/kg. Twenty-four hours later, the bronchoalveolar fluid, peripheral blood, and lungs were collected for cellular and molecular analysis. The milonine treatment decreased the cell migration (mainly neutrophils) to the alveoli, the pulmonary edema, and the cytokine levels (IL-1ß, IL-6, TNF-α). The systemic IL-6 level was also reduced. The milonine docking analysis demonstrated hydrophobic interaction at TLR4/MD2 groove with Ile124 and Phe126 amino acids. Indeed, the alkaloid downregulated the kinase-Akt and NF-κB through TLR4/MD2. Therefore, milonine is an effective inflammatory modulator being a potential molecule for the treatment of lung inflammation.

Leishmania (L.) infantum BH401 strain induces classic renal lesions in dogs: Histological and confocal microscopy study.

Extracellular matrix (ECM) alterations in visceral leishmaniasis are related mainly to collagen deposition (fibropoiesis). In canine visceral leishmaniasis (CVL), an intense fibrosis associated to chronic inflammation in organs such as kidneys is described. However, renal fibropoiesis has not been described in natural or experimental infections with L. (L.) infantum. We aimed to characterize renal nephropathies by histology and confocal microscopy comparing renal lesions in dogs naturally and experimentally infected with L. (L.) infantum. Sixty-two mixed-breed symptomatic dogs naturally infected with L. (L.) infantum, sixteen beagles experimentally infected with two strains of L. infantum (eleven dogs with the BH400 strain and five dogs with the BH401 strain), and five uninfected beagles (controls) were used. Samples were stained with hematoxylin & eosin for routine histology. Congo red was used to visualize amyloid protein deposits, periodic acid-Schiff to identify glomerular basal membrane anomalies, Masson's trichrome for collagen deposits, and Jones' methenamine silver to reveal membranous glomerulonephropathy. Immunohistochemistry was used to identify Leishmania amastigotes, and confocal microscopy was used for macrophage characterization (L1/calprotectin and CD163 antigen receptors). The most common lesions were chronic glomerular and interstitial nephritis, which was found in all naturally infected dogs and dogs experimentally infected with L. infantum strain BH401 but not with the BH400 strain. Glomeruloesclerosis was the main lesion presented in all BH401 group. Morphometric analysis revealed positive correlation of renal glomeruli tufts with cellular expression of L1/calprotectin and CD163 antigens. Leishmania infantum strain BH401 shows pathogenicity that may be sufficient to induce classic chronic visceral renal leishmaniasis.

Functional and morphologic dysfunctions in the airways of rats submitted to an experimental model of obesity-exacerbated asthma.

The obesity-exacerbated asthma phenotype is characterized by more severe asthma symptoms and glucocorticoid resistance. The aim of this study was to standardize an obesity-exacerbated asthma model by a high glycemic level index (HGLI) diet and ovalbumin (OVA) sensitization and challenges in Wistar rats. Animals were divided into groups: control (Ctrl), obese (Ob), asthmatic (Asth), obese asthmatic (Ob + Asth) and obese asthmatic treated with dexamethasone (Ob + Asth + Dexa), and in vivo and in vitro functional and morphological parameters were measured. After HGLI consumption, there was an increase in body weight, fasting blood glucose, abdominal circumferences, body mass index and adiposity index. Respiratory function showed a reduction in pulmonary tidal volume and ventilation. In isolated tracheas, carbachol showed an increase in contractile efficacy in the Ob, Ob + Asth and Ob + Asth + Dexa, but mostly on Ob + Asth. Histological analysis of lungs showed peribronchovascular inflammation and smooth muscle hypertrophy and extracellular remodeling on Ob + Asth and Ob + Asth + Dexa. An obesity-exacerbated asthma model was successfully established. Therefore, this model allows further molecular investigations and the search for new therapies for the treatment and relief of symptoms of patients with obesity-induced resistant asthma.

Ouabain modulates airway remodeling caused by Th2-high asthma in mice.

OBJECTIVE: Ouabain, an inhibitor of Na+/K+-ATPase, is a type of endogenous hormone synthesized in the adrenal cortex and hypothalamus. Previous studies found that ouabain potently inhibited acute inflammatory reactions such as type 2 inflammation and regulated immunological processes. In this study, we aimed to investigate ouabain effect on allergic asthma. METHODS: BALB/c mice were submitted to chronic airway allergic inflammation induced by an ovalbumin (OVA) protocol. The animals were treated with ouabain or standard drug, budesonide. The following parameters were evaluated: cell migration, cytokine profile, IgE levels, lung histological modifications and MAPK activation. RESULTS: At first, it was observed that ouabain reduced OVA-induced cell migration into the lung, observed by bronchoalveolar lavage fluid (BALF) cell counting and lung histological analysis (HE stain). Additionally, ouabain negatively modulated alarmins (IL-33 and TSLP), Th2 high cytokines levels (IL-1ß and IL-4) and tissue remodeling markers such as TNF-α and TGF-ß. Treatment with ouabain also reduced OVA-specific IgE titers in BALF and serum, respectively, when compared to the OVA group. Lung histological parameters, including collagen deposition and mucus production induced by OVA were also attenuated by ouabain treatment. Finally, our results showed that p38 mitogen-activated protein kinase (MAPK) signaling pathways were suppressed by ouabain in this model. All these parameters were reduced by budesonide, a steroidal anti-inflammatory standard drug. CONCLUSION: These data together suggest that, in addition to its acute anti-inflammatory action, ouabain is also able to modulate allergic asthma.

Malay apple (Syzygium malaccense) promotes changes in lipid metabolism and a hepatoprotective effect in rats fed a high-fat diet.

This study aimed to investigate the effects of Malay apple (MA) on the glycaemic response, lipid metabolism and liver health of rats fed a normal- or high-fat diet. Male Wistar rats were initially randomized into two groups: normal-fat diet (NF, n = 16) and high-fat diet (HF, n = 16) for three weeks. Then, they were subdivided into: normal-fat diet (NF, n = 8); normal-fat diet supplemented with MA (NFMA, n = 8); high-fat diet (HF, n = 8); high-fat diet supplemented with MA (HFMA, n = 8), continuing for another three weeks with their respective diets. Simultaneously, the supplemented groups received MA (400 mg/kg body weight) via gavage. MA contains anthocyanins (26.59 mg/100 g), catechin (2.55 ± 0.33 mg/100 g) and gallic acid (1.07 ± 0.20 mg/100 g) as the main phenolics. MA had a low-key effect on insulin and glucose tolerance, however decreased serum lipids and protected the liver from steatosis induced by the high-fat diet. HFMA rats showed changes in the fatty acid composition of the liver and faeces and decreased liver cholesterol levels (20.5%); as well as increased faecal excretion of fat (98%), cholesterol (10.5%) and bile acids (42.2%) when comparing to HF. MA supplementation reduced food and fat intake in HFMA in the last two weeks of the experiment, which may also have influenced these results. Our results suggest that MA was not able to improve glycaemic parameters, however, had hepatoprotective effect and minimizes changes in lipid metabolism caused by high-fat diet intake, which can relate mainly to the phenolic compounds present in this fruit.

Evaluation of Toxicity and Oxidative Stress of 2-Acetylpyridine-N(4)-orthochlorophenyl Thiosemicarbazone.

Thiosemicarbazones are well known for their broad spectrum of action, including antitumoral and antiparasitic activities. Thiosemicarbazones work as chelating binders, reacting with metal ions. The objective of this work was to investigate the in silico, in vitro, and in vivo toxicity and oxidative stress of 2-acetylpyridine-N(4)-orthochlorophenyl thiosemicarbazone (TSC01). The in silico prediction showed good absorption by biological membranes and no theoretical toxicity. Also, the compound did not show cytotoxicity against Hep-G2 and HT-29 cells. In the acute nonclinical toxicological test, the animals treated with TSC01 showed behavioral changes of stimulus of the central nervous system (CNS) at 300 mg/kg. One hour after administration, a dose of 2000 mg/kg caused depressive signs. All changes disappeared after 24 h, with no deaths, which suggest an estimated LD50 of 5000 mg/kg and GSH 5. The group treated with 2000 mg/kg had an increase of water consumption and weight gain in the second week. The biochemical parameters presented no toxicity relevance, and the analysis of oxidative stress in the liver found an increase of lipid peroxidation and nitric oxide. However, histopathological analysis showed organ integrity was maintained without any changes. In conclusion, the results show the low toxicological potential of thiosemicarbazone derivative, indicating future safe use.

Safety and tolerability of cinnamaldehyde in orabase for oral candidiasis treatment: phase I clinical trial.

OBJECTIVE: To advance studies on the effect of a new pharmaceutical formulation for the treatment of oral fungal infections, we evaluated the safety and tolerability of orabase ointment containing cinnamaldehyde for use on the oral mucosa. MATERIAL AND METHODS: A clinical trial (phase I) was carried out on 35 individuals with healthy oral mucosa divided into three groups: ointments at 200 µg/mL, n = 12; 300 µg/mL, n = 11; and 400 µg/mL, n = 12. Product safety was assessed using three parameters: (a) clinical evolution as recorded by trained examiners; (b) evolution of the inflammatory process as registered by an exfoliative cytology exam and analyzed by trained pathologists; (c) mucosal swab to count Candida spp. colony-forming units (CFU). These parameters were analyzed both beforehand and at 15 days of treatment. RESULTS: The three ointment concentrations evaluated did not trigger inflammatory processes. The mycological analyses revealed a reduction of at least 99% in the number of Candida spp. CFU. In the exfoliative cytology analyses, the cells were found to be healthy. Participants reported a pleasant taste, yet 17% reported a slight burning sensation when applying the product. CONCLUSIONS: The ointment is safe and tolerable for use on healthy oral mucosa. TRIAL REGISTRATION: Registration number: RBR-7zwzs3. CLINICAL RELEVANCE: The ointment proved to be safe and tolerable for use on oral mucosa, encouraging studies to evaluate its clinical efficacy in patients with oral candidiasis, and contributing to a new therapeutic proposal for the treatment of fungal infections caused by Candida spp.

4-Carvomenthenol, a monoterpene of essential oils, and its underlying effects on anti-inflammatory activity and immediate hypersensitivity reaction

Abstract The monoterpene 4-carvomenthenol (Carvo) is found in essential oils of plant. Here, we evaluate the Carvo oral pretreatment in acute inflammatory experimental models and in silico molecular docking. Mice pretreated with Carvo were challenged and submitted to the protocols: paw edema, peritonitis, scratching behavior and anaphylactic shock reaction. Besides, we used histamine H1 receptor, cyclooxygenases (COX-1 and COX-2) and phospholipase A2, as targets for molecular docking analysis. Carvo inhibited the carrageenan-induced paw edema and decreased the peritoneal influx of polymorphonuclear cells on carrageenan-challenged mice without interfering with the mononuclear cell influx. Moreover, Carvo diminished the histamine, PGE2 and compound 48/80 induced paw edematogenic effect. The monoterpene also diminished the mice scratching behavior and, surprisingly, avoided the animal death caused by compound 48/80 in 30 min. Through the docking analysis, Carvo showed favorable binding energy to the histamine H1 receptor. This study demonstrates that Carvo attenuated the allergic inflammatory process, decreasing edema, cell migration, activation of mast cells and the histamine release, probably due to interaction of Carvo with the histamine H1 receptor, ameliorating the itching and the anaphylactic shock reaction. Therefore, the results of this study indicate that Carvo has anti-inflammatory properties by reducing the histamine effects.