ß-Cyclodextrins alter the energy metabolism-related enzyme activities in rats.

Although widely used in medicine, separation technology, and other fields, the effects of cyclodextrins on the activities of phosphoryl transfer enzymes have not been previously evaluated. In vivo studies evaluated the function of cyclodextrins as active compounds. Despite the use of cyclodextrins as active compounds, the effects of cyclodextrins on hepatic and renal tissues remain to be fully elucidated. The primary objective of this study was to evaluate the effects of ß- cyclodextrins, methyl-ß-cyclodextrin (M-ß- cyclodextrins), and (2-hydroxypropyl)-ß-cyclodextrin (HP-ß-cyclodextrins) on enzyme activities regulating the maintenance of energy homeostasis in the kidney and liver tissues in relation to toxicity. Serum levels of liver and kidney markers were measured, and oxidative stress parameters were assessed. After 60-day treatments, we observed that the administration of ß-cyclodextrins and M-ß-cyclodextrins inhibited the hepatic activity of pyruvate kinase, an irreversible enzyme within the glycolytic pathway. Additionally, administration of HP-ß-cyclodextrins inhibited creatine kinase activity and increased the total sulfhydryl content in kidneys. Here, we demonstrated for the first time that ß-cyclodextrins, M-ß-cyclodextrins, and HP-ß-cyclodextrins cause bioenergetic dysfunction in renal and hepatic tissues. These findings suggest that understanding the balance between cyclodextrins' efficacy and adverse effects is essential for better accepting their use in medicine.

MicroRNA qPCR normalization in Nile tilapia (Oreochromis niloticus): Effects of acute cold stress on potential reference targets.

The Nile tilapia (Oreochromis niloticus) is one of the most important cultured fish worldwide, but tilapia culture is largely affected by low temperatures. Recent studies suggest that microRNAs (miRNAs) regulate cold tolerance traits in fish. In general, qPCR-based methods are the simplest and most accurate forms of miRNA quantification. However, qPCR data heavily depends on appropriate normalization. Therefore, the aim of the present study is to determine whether the expression of previously tested, stably expressed miRNAs are affected by acute cold stress in Nile tilapia. For this purpose, one small nuclear RNA (U6) and six candidate reference miRNAs (miR-23a, miR-25-3, Let-7a, miR-103, miR-99-5, and miR-455) were evaluated in four tissues (blood, brain, liver, and gills) under two experimental conditions (acute cold stress and control) in O. niloticus. The stability of the expression of each candidate reference miRNA was analyzed by four independent methods (the delta Ct method, geNorm, NormFinder, and BestKeeper). Further, consensual comprehensive ranking of stability was built with RefFinder. Overall, miR-103 was the most stable reference miRNA in this study, and miR-103 and Let-7a were the best combination of reference targets. Equally important, Let-7a, miR-23a, and miR-25-3 remained consistently stable across different tissues and experimental groups. Considering all variables, U6, miR-99-5, and miR-455 were the least stable candidates under acute cold stress. Most important, suitable reference miRNAs were validated in O. niloticus, facilitating further accurate miRNA quantification in this species.

Chronic cold exposure modulates genes related to feeding and immune system in Nile tilapia (Oreochromis niloticus).

Nile tilapia is the fourth most produced species in the global aquiculture panorama. This species requires water temperatures higher than 16 °C to grow and survive, and so, little is known about the effects of low temperatures on genes related to food intake and inflammatory responses. This study brought insights about the modulation of genes in different tissues of Nile tilapia chronically exposed to low temperatures. Thus, sixty animals were divided in two experimental groups: a control group in which the animals remained at the optimum temperature of 24 °C; and an exposed to cold group, in which a decrease in the water temperature was applied until reaching 15 °C. These conditions were maintained for 28 days. Blood samples were collected for flow cytometry analysis, while brain, spleen, liver, and kidney tissues were collected for total RNA extraction, followed by quantitative PCR (RT-qPCR). For genes related to feeding process pathway, it was observed an upregulation in pyy and a downregulation of npy and cart gene expression. Also, pro-inflammatory cytokine genes were modulated in the spleen, kidney and liver with a higher expression of il-1b and tnfα and a reduction in the il-8 and nf-κß gene expressions in the group exposed to 15 °C. The fish exposed to cold presented higher serum cortisol levels than the ones from control group. The blood cell analysis showed a lower level of membrane fluidity and a higher DNA fragmentation and cell disruption in the group exposed to cold. These findings suggest an important effect of a stressful situation in the tilapia organism due to cold exposure. This study brings insights on tilapia wellbeing under low temperature stress. It can be a first step to understanding the appropriate way to cope with cold impacts on aquaculture.

Resveratrol and resveratrol-hydroxypropyl-ß-cyclodextrin complex recovered the changes of creatine kinase and Na+, K+-ATPase activities found in the spleen from streptozotocin-induced diabetic rats.

Type 1 diabetes (T1D) is the result of the selective destruction of the pancreatic ß-cells by T cells of the immune system. Although spleen is a secondary lymphoid organ, it is also involved in the T1D pathogenesis. However, the alterations in a variety of cellular processes of this disease need to be further understood. We aimed to analyze the benefits of resveratrol, and its complexed form on diabetic complications in the spleen of rats. To this end, we investigated important enzymes of phosphoryl transfer network, and Na+, K+-ATPase activity. Wistar rats were divided into non-diabetic groups: Control, Ethanol, Resveratrol, Hydroxypropyl-ß-cyclodextrin, Resveratrol-hydroxypropyl-ß-cyclodextrin, and diabetic groups with the same treatments. Diabetes was induced by a single dose of 60 mg/kg of streptozocin intraperitoneally, and treatments by intragastric gavage once daily for 60 days. Hyperglycemia reduced creatine kinase activity, which was reversed by the administration of resveratrol. Na+, K+-ATPase activity was greatly affected, but it was reversed by resveratrol and resveratrol-hydroxypropyl-ß-cyclodextrin. This suggest an energetic imbalance in the spleen of diabetic rats, and in case this also occurs in the diabetic patients, it is possible that resveratrol supplementation could be beneficial to the better functioning of the spleen in diabetic patients.

Experimental infection of cattle with Listeria monocytogenes: Participation of purinergic metabolism in disease pathogenesis.

The objective of this study was to evaluate whether experimental infection with Listeria monocytogenes alters the activity of triphosphate diphosphohydrolase (NTPDase), 5'-nucleotidase, and adenosine deaminase (ADA) in cattle. Ten male Holstein breed cattle were divided in two groups of five animals each: a control group, and a group infected with a pathogenic strain of L. monocytogenes. We drew blood for platelets on days 0, 7 and 14 of the experiment. On the 14th day post infection (PI), the animals were euthanized. Brain, spleen and liver were processed for histopathological examination and measurement of enzyme activities. The five (n = 5/5) bovines experimentally infected by L. monocytogene were positive-PCR in hepatic tissue. In the brain, only four (n = 4/5) of these animals were positive-PCR for listeriosis. There were no differences in platelet counts between groups (P > 0.05). In platelets, NTPDase activity (with ATP and ADP as substrates) were higher on the 7th PI day in the infected group, whereas the activities of 5'-nucleotidase and ADA were higher on the 7th and 14th PI. In serum and liver, ADA activity was higher in infected animals, but was lower on day 14 PI in spleen. NTPDase activity (with ATP as substrate) was higher in the cerebellum of infected animals, but was lower in the cerebral cortex and medulla oblongata. NTPDase activity (with ADP as substrate) was lower in the cerebellum and cerebral cortex of infected animals, whereas 5'-nucleotidase was higher. ADA activity was lower in the cerebellum, cerebral cortex and medulla oblongata in infected animals compared with controls. In conclusion, there appears to be a protective immunomodulatory response in spleen and brain structures of cattle infected with L. monocytogenes.

Listeria monocytogenes impairs enzymes of the phosphotransfer network and alters antioxidant/oxidant status in cattle brain structures.

Several evidences have suggested the involvement of enzymes belonging to the phosphotransfer network, formed by creatine kinase (CK), pyruvate kinase (PK) and adenylate kinase (AK), as well the oxidative stress on the pathogenesis of infectious diseases associated with the central nervous system (CNS). Thus, the aim of this study was to evaluate whether listeriosis alters the brain energy metabolism and/or causes oxidative stress in different brain structures of cattle experimentally infected by Listeria monocytogenes. The cytosolic CK activity was inhibited in the cerebral cortex, cerebellum, brainstem and hippocampus of infected animals compared to uninfected animals, while the mitochondrial CK activity was increased. The PK activity was inhibited in all brain structures of infected animals, while the AK activity was unchanged. Na+, K+-ATPase activity decreased in the cerebral cortex, cerebellum and hippocampus of animals infected by L. monocytogenes. Regarding the oxidative strees variables, the cerebellum and brainstem of infected animals showed increased thiobarbituric acid reactive substances, while the catalase activity was inhibited. Glutathione S-transferarase was inhibited in the cerebral cortex and brainstem of infected animals, and it was increased in the cerebellum. L. monocytogenes was quantified in the liver (n = 5/5) and cerebral cortex (n = 4/5) of the infected cattle. Based on these evidences, the nucleocytoplasmic communication between CK isoenzymes was insufficient to avoid an impairment of cerebral bioenergetics. Moreover, the inhibition on brain PK activity caused an impairment in the communication between sites of ATP generation and ATP utilization. The lipid peroxidation and alteration on antioxidant status observed in some brain structures were also involved during the disease. In summary, these alterations contribute to disease pathogenesis linked to CNS during cattle listeriosis.

Trypanocidal activity of the compounds present in Aniba canelilla oil against Trypanosoma evansi and its effects on viability of lymphocytes.

Aniba canelilla (H.B.K.) Mez, popularly known as "casca-preciosa" (precious bark), is a plant of the Lauraceae family, widely distributed in the Amazon region. Its major constituent is 1-nitro-2-phenylethane, a rare molecule in plants which is responsible for this plant's cinnamon scent. The present study aimed to report the chemical characterization of the oil extracted from Aniba canelilla using gas-chromatography/mass spectrometry and to assess its in vitro trypanocidal activity against Trypanosoma evansi, a prevalent haemoflagellate parasite that affects a broad range of mammal species in Africa, Asia and South America. The oil presented 1-nitro-2-phenylethane (83.68%) and methyleugenol (14.83%) as the two major components. The essential oil as well as both major compounds were shown to exert trypanocidal effect. Methyleugenol was slightly more active than 1-nitro-2-phenylethane. In vitro studies showed that the oil extracted from the stems of A. canelilla may be regarded as a potential natural treatment for trypanosomosis, once proven their in vivo action, may be an interesting alternative in the treatment of infected animals with T. evansi.

Effect of senolytic drugs in young female mice chemically induced to estropause.

AIMS: This study aimed to assess metabolic responses and senescent cell burden in young female mice induced to estropause and treated with senolytic drugs. MAIN METHODS: Estropause was induced by 4-vinylcyclohexene diepoxide (VCD) injection in two-month-old mice. The senolytics dasatinib and quercetin (D + Q) or fisetin were given by oral gavage once a month from five to 11 months of age. KEY FINDINGS: VCD-induced estropause led to increased body mass and reduced albumin concentrations compared to untreated cyclic mice, without affecting insulin sensitivity, lipid profile, liver enzymes, or total proteins. Estropause decreased catalase activity in adipose tissue but had no significant effect on other redox parameters in adipose and hepatic tissues. Fisetin treatment reduced ROS levels in the hepatic tissue of estropause mice. Estropause did not influence senescence-associated beta-galactosidase activity in adipose and hepatic tissues but increased senescent cell markers and fibrosis in ovaries. Senolytic treatment did not decrease ovarian cellular senescence induced by estropause. SIGNIFICANCE: Overall, the findings suggest that estropause leads to minor metabolic changes in young females, and the senolytics D + Q and fisetin had no protective effects despite increased ovarian senescence.

Trypanocidal activity of the essential oils in their conventional and nanoemulsion forms: in vitro tests.

The aim of this study was to investigate the susceptibility in vitro of Trypanosoma evansi to the essential oils of andiroba (Carapa guaianensis) and aroeira (Schinus molle), in their conventional and nanostructured forms. For that, pure oils at concentrations of 0.5%, 1.0% and 2.0% were used. A negative control (untreated) and a positive control (diminazene aceturate 0.5%) were used as comparative parameters. Later, the same tests were performed, using nanoemulsions oils at concentrations of 0.5% and 1.0%. The tests were carried out in triplicates and the numbers of parasites were quantified on 1, 3 and 6 h from onset of the study. A dose-dependent reduction in the number of parasites to the forms of two oils tested was observed after 1 h. The concentration of parasites was significantly reduced at low concentrations after 3 h, as well as at 6 h no alive parasites were observed for the essential oils tested. Ours findings indicate, for the first time, that oils of andiroba and aroeira (in their conventional and nanoemulsion forms) have high activity against T. evansi in vitro, leading to the suggestion that these oils may be applied as an alternative treatment for this disease.

Pharmacological modulation of Na+, K+-ATPase as a potential target for OXA-induced neurotoxicity: Correlation between anxiety and cognitive decline and beneficial effects of 7-chloro-4-(phenylselanyl) quinoline.

Growing evidence demonstrates that Oxaliplatin (OXA) is commonly associated with neurotoxicity that leads to emotional and cognitive impairments. The aim of the present study was to evaluate the OXA and Na+, K+-ATPase interaction and to correlate anxious behavior and cognitive impairment induced by this chemotherapeutic in Swiss mice. Also, considering the pharmacological modulation of Na+, K+-ATPase as a potential target for OXA-induced neurotoxicity, the therapeutic potential of 7-chloro-4-(phenylselanyl) quinoline (4-PSQ) was evaluated. Mice received OXA (10 mg kg-1) or vehicle by intraperitoneal route (days 0 and 2). Oral administration of 4-PSQ (1 mg kg-1) or vehicle was performed from days 2-14. Behavioral tasks started from day 12 onwards. On day 15, the animals were sacrificed, and the tissues collected. The effects of OXA and 4-PSQ on activity and expression level of Na+, K+-ATPase in the hippocampus and cerebral cortex, and the plasmatic corticosterone levels were determined. The findings demonstrated a significant positive correlation between anxious behavior and cognitive impairment induced by OXA. OXA caused an increase on the plasmatic corticosterone levels and reduced activity and expression level of Na+, K+-ATPase. 4-PSQ reduced both anxious behavior and cognitive impairment induced by OXA. 4-PSQ effect seems to be due to the modulation of Na+, K+-ATPase and reduction of corticosterone levels. Our results helped to expand knowledge about the mechanisms involved in the physiopathology of the OXA-induced neurotoxicity and strongly indicated that 4-PSQ may be a good prototype for the treatment of anxious behavior and cognitive impairment induced by OXA exposure.

Experimental listeriosis: A study of purinergic and cholinergic inflammatory pathway.

The cholinergic, purinergic and oxidative stress systems were related to nervous system damage in some pathologies, as well as being involved in pro-inflammatory and anti-inflammatory pathways. The objective was to investigate changes in purinergic, cholinergic systems and oxidative stress related to the neuropathology of listeriosis. Gerbils were used as experimental models. The animals were divided in two groups: control and infected. The animals were orally infected with 5 × 108 CFU/animal of the pathogenic strain of Listeria monocytogenes. Collected of material was 6 and 12th days post-infection (PI). Infected animals showed moderate mixed inflammatory infiltrates in the liver. The spleen and brain was used for PCR analyses, confirming infection by L. monocytogenes. Increase in number of total leukocytes because of an increase in lymphocytes in infected (P < 0.001). ATP and ADP hydrolysis by NTPDase was lower at 6 and 12th days PI in infected animals than in the control group. ADA (adenosine deaminase) activity was higher on the 6th day PI (P < 0.05) and decreased on the 12th day PI (P < 0.05) in infected animals. AChE (acetylcholinesterase) activity did not differ between groups on the 6th day PI; however, activity decreased in infected group on the 12th day PI (P < 0.05). On the 12th day PI, an increase of oxygen-reactive species levels and lower catalase and superoxide dismutase activities in the infected group was observed, characterizing a situation of cerebral oxidative stress. The inflammatory and oxidative mechanisms are present in listeriosis in asymptomatic animals, and that ectonucleotidases and cholinesterase's are involved in immunomodulation.

Polysaccharide-based film loaded with vitamin C and propolis: A promising device to accelerate diabetic wound healing.

Wound healing can be a painful and time-consuming process in patients with diabetes mellitus. In light of this, the use of wound healing devices could help to accelerate this process. Here, cellulose-based films loaded with vitamin C (VitC) and/or propolis (Prop), two natural compounds with attractive properties were engineered. The starting materials and the cellulose-based films were characterized in detail. As assessed, vitamin C can be released from the Cel-PVA/VitC and Cel-PVA/VitC/Prop films in a controlled manner. In vitro antibacterial activity studies showed a reduction of bacteria counts (Escherichia coli and Staphylococcus aureus) after Cel-PVA/VitC, Cel-PVA/Prop, and Cel-PVA/VitC/Prop treatments. Moreover, we examined the antibacterial and wound healing properties of the cellulose-based films in a streptozotocin (STZ)-induced diabetic animal model. Diabetic mice exhibited impaired wound healing while the Cel-PVA/VitC/Prop treatment increased the wound closure. A marked reduction in bacterial counts present in the wound environment of diabetic mice was observed after Cel-PVA/VitC, Cel-PVA/Prop and Cel-PVA/VitC/Prop treatment. Histological analysis demonstrated that the non-treated diabetic mice group did not exhibit adequate wound healing while the treated group with Cel-PVA/VitC and Cel-PVA/VitC/Prop films presented good cicatricial response. Furthermore, these novel eco-friendly films may represent a new therapeutic approach to accelerate diabetic wound healing.

In vitro effect of seven essential oils on the reproduction of the cattle tick Rhipicephalus microplus.

The acaricidal effect of seven essential oils was examined in vitro against the cattle tick (Rhipicephalus microplus). Engorged female ticks were manually collected in farms of Southern Brazil and placed into petri dishes (n = 10) in order to test the following oils: juniper (Juniperus communis), palmarosa (Cymbopogon martinii), cedar (Cedrus atlantica), lemon grass (Cymbopogon citratus), ginger (Zingiber officinale), geranium (Pelargonium graveolens) and bergamot (Citrus aurantium var bergamia) at concentrations of 1%, 5%, and 10% each. A control group was used to validate the tests containing Triton X-100 only. Treatment effectiveness was measured considering inhibition of tick oviposition (partial or total), egg's weight, and hatchability. C. martinii, C. citratus and C. atlantica essential oils showed efficacy higher than 99% at all concentrations tested. In addition, J. communis, Z. officinale, P. graveolens, and C. aurantium var bergamia oils showed efficiency ranging from 73% to 95%, depending on the concentration tested, where higher concentrations showed greater efficacy. It was concluded that essential oils can affect tick reproduction in vitro by inhibiting oviposition and hatchability.

Using of essential oils in the treatment of mice infected with Trypanosoma evansi / Uso de aceites esenciales en el tratamiento de ratones infectados con Trypanosoma evansi

Objective. This study aimed to test the effectiveness of copaiba, andiroba and aroeira essential oils for controlling trypanosomosis by Trypanosoma evansi with mice as experimental model. Materials and methods. Sixty-six mice were divided into eleven groups (A to L) with six animals each. Group A was the unique composed by healthy and uninfected animals (negative control). Animals in groups B to L were inoculated with 0.1 mL of blood containing 2.7 x 10(6) trypanosomes. Group B was used as a positive control without treatment. In experiment were tested copaiba (C, D and E), andiroba (F, G and H) and aroeira (I, J and L) oils at doses of 0.6, 0.8 and 1.0 mL kg-1 to infected mice (T. evansi). Results. These protocols did not provide curative efficacy; however, the mice treated with highest dose of copaiba showed a significant increase in the longevity when compared others groups. Conclusions. Previously in our studies, these essential oils have shown trypanocidal activity in vitro, but when they were tested in vivo in mice infected with T. evansi, this trypanocidal activity, or the curative effect was not found, being only able to prolong the lifespan of the animals treated with copaiba oil.

Objetivo. Este estudio tuvo como objetivo evaluar la eficacia de los aceites esenciales de copaiba, andiroba y aroeira para controlar la tripanosomiasis por Trypanosoma evansi con ratones como modelo experimental. Materiales y métodos. Sesenta y seis ratones se dividieron en once grupos (A a L) con seis animales cada uno. Grupo A fue el único compuesto por los animales sanos y no infectadas (control negativo). Los animales en los grupos B a L fueron inoculados con 0,1 mL de sangre que contiene 2,7 x 10(6) tripanosomas. Grupo B se utilizó como control positivo, sin tratamiento. En el experimento se pusieron a prueba los aceites de copaiba (C, D y E), andiroba (F, G y H) y aroeira (I, J y L) en una dosis de 0,6, 0,8 y 1,0 ml kg-1 en ratones infectados (T. evansi). Resultados. Estos protocolos no proporcionan una eficacia curativa; sin embargo, los ratones tratados con la dosis más alta de copaiba mostraron un aumento significativo en longevidad en comparación con otros grupos. Conclusiones. De forma previa en nuestros estudios, estos aceites esenciales han demostrado actividad tripanocida in vitro, pero cuando se ensayaron in vivo en ratones infectados con T. evansi, no se encontró esta actividad tripanocida o el efecto curativo, siendo sólo capaz de prolongar la vida de los animales tratados con aceite de copaiba.