Sheep naturally infected with Anaplasma ovis - evaluation of cardiac and inflammatory biomarkers

Background: Anaplasmosis, also called gall sickness or tropical bovine ehrlichiosis, is an infectious disease caused by species belonging to the genus Anaplasma in domestic and wild animals in tropical and subtropical regions. Anaplasma ovis and A. phagocytophilum are important pathogens of sheep. A. ovis is considered the most common species affecting sheep. The infection is usually subclinical and progresses with high fever, anaemia, icterus, weight loss and abortions. This study aimed to investigate changes in cardiac damage markers, oxidative stress and antioxidant status, cytokines, and acute phase proteins in sheep naturally infected with A. ovis. Materials, Methods & Results: For this purpose, a total of 40 animals, including 20 healthy sheep and 20 sheep infected with anaplasmosis, were used. A. ovis was diagnosed based on clinical findings and peripheral blood smear. Blood smears were prepared from the ear vein. The smears were stained with Giemsa and examined for the presence of Anaplasma spp. Infection was also confirmed by polymerase chain reaction (PCR) analysis. The genomic DNA was isolated from blood, and the MSP-4 gene region was amplified as A. ovis specific target gene. Twenty clinically healthy sheep of the same age group, reared under the same conditions and testing negative in the molecular assessment were used as controls. Blood samples were collected from the cephalic vein and and centrifuged to obtain serum. The serum stored at -20°C until the analysis stage. Serum samples were used for the analysis of cardiac damage markers [troponin I (cTnI), creatine kinase MB (CK-MB), lactate dehydrogenase (LDH) and aspartate transaminase (AST)], oxidative stress parameters [malondialdehyde (MDA), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], cytokines [interleukins IL-6, IL-1ß and IL-10, tumour necrosis factor α (TNF-α), and interferon-γ (IFN-γ)] and acute phase proteins [C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp)]. cTnI and CK-MB levels were measured using a chemiluminescent immunoassay. MDA, TAS, SOD, CAT, GPx, TNF-α, IL-1ß, IL-6, IL-10, IFN-γ, SAA and Hp levels were measured by an ELISA reader. LDH, AST and CRP levels were measured in an autoanalyzer. cTnI and LDH levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The concentration of AST was decreased in infected animals. MDA, TAS, SOD, CAT and GPx levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The levels of the inflammatory parameters such as TNF-α, IL-1ß, IL-10 and IFN-γ were significantly increased in the infected animals compared to the healthy ones (P < 0.05). Hp level were significantly increased in the infected group compared with the control group (P < 0.05). However, there was no significant change in CK-MB, SAA and CRP concentrations in the infected animals (P > 0.05). Discussion: Ovine anaplasmosis is an obligate intracellular arthropod disease that causes widespread changes in haematobiochemical, immune response and oxidative stress parameters. Cardiac damage is often overlooked in field conditions due to the lack of adequate knowledge about the pathophysiology of the disease. Our results showed that A. ovis infection leads to significant changes in cardiac biomarkers and that the parasite can cause cardiac dysfunction. This is the first report on cardiac damage markers in Anaplasma-infected sheep. Additionally, the levels of proinflammatory and oxidative stress markers that may cause functional disorders were also found to be increased. Thus, measuring markers of cardiac function, oxidative stress and inflammation can be a useful tool in the early diagnosis of ovine anaplasmosis.

Licorice zinc suppresses melanogenesis via inhibiting the activation of P38MAPK and JNK signaling pathway in Balb/c mice skin

Purpose: The active melanocytes in the skin were affected by hormones and ultraviolet (UV) irradiation. Licorice zinc has a whitening effect, which may have a prominent potential in the treatment of pigmented skin disease. Methods: Modeling chloasma C57BL/6J mice by daily progesterone injection (15 mg/kg) and ultraviolet B (UVB) irradiation (λ = 312 nm, 2 h/day) for 30 days. Then, mice were given 0.65, 1.3, and 2.6 (g/kg) of licorice zinc and tranexamic acid 250 mg daily by oral administration for 14 days, respectively. Hematoxylin and eosin and Fontana-Masson staining, and Western blotting (WB) were performed to test the inhibitory of melanogenesis and activation of c-Jun-N-terminal (JNK)/p38 mitogen-activated protein kinases (MAPK) for licorice zinc. Melanogenesis was induced by α-melanocyte-stimulating hormone in vitro. Cell counting kit-8, melanin content determination, and WB were performed to verify the inhibitory effect of licorice zinc on melanogenesis. Results: The present study showed that licorice zinc decreased melanin formation, cutaneous tissue injury, and the phosphorylation of JNK and P38MAPK, which was caused by UVB irradiation in vivo. In vitro, licorice zinc showed opposite effects from JNK/p38 activator. Meanwhile, tyrosinase-related protein-1, tyrosinase, and microphthalmia-associated transcription factor were decreased too. Conclusions: Licorice zinc induced a decrease in melanin synthesis by inhibiting the JNK and the P38MAPK signaling pathway, suggesting licorice zinc is a potential agent of anti-chloasma.

Canine mast cell tumour lomustine's sensitization by tyrosine kinase inhibitors

Background: Tyrosine kinase inhibitors (TKIs) may sensitize neoplasms to conventional antineoplastic agents, however such studies are scarse in the veterinary literature and there is no in vivo study about this subject. Although the literature recommend consensual about the use of masitinib for unresectable or metastatic MCTs, the potential of tumour sensitization to chemotherapeutic agents exerted by the drug is poorly explored in veterinary medicine. The objective of this paper was to report, for the first time, the sensitization of 2 canine mast cell tumours (MCTs) to lomustine, with the use of 2 tyrosine kinase inhibitors: masitinib and toceranib. Cases: Two dogs were referred due tumour recurrence in the left pelvic limb (dog 1), and unilateral mass in the right nasal mucocutaneous region (dog 2). The first case was a 8-year-old female Pinscher, and the second case refers to a 8-year-old male mixed-breed dog. Fine needle aspiration of both lesions was performed, and the cytological analysis were compatible with high grade canine MCT. In the first case, it was started a chemotherapeutic treatment with intravenous vinblastine (2 mg/m² ), associated with prednisolone (40 mg/m2 , every 24 h for 7 days), followed by 25 mg/m2 every 24 h, for more 30 days, tramadol (4 mg/kg every 8 h, until new recommendations) and gabapentin (3 mg/kg every 12 h, until new recommendations). However, there was no objective response, and vinblastine was substituted by lomustine (60 mg/m2 every 21 days), however there was also no response after 2 doses. After masitinib importation, the same was started at 12.5 mg/kg orally every 24 h, but there was also no objective response. However, after new lomustine administration the lesion showed complete remission. The second dog initiated its treatment with toceranib, recently licensed in Brazil, at a dosage of 2.7 mg/kg every 48 h, and after 30 days, there was partial remission. However, the remaining lesion still deemed unresectable, and systemic chemotherapy with lomustine (50 mg/m2 ) was initiated along with continuous toceranib. After 3 weeks of the first chemotherapy complete remission was noted and a second dose was administered. Once the patient remained in complete clinical remission, only toceranib was maintained at the same dose. After 11 months using the toceranib, there was sign of disease recurrence and lomustine was re-initiated resulting in complete remission. Discussion: The TKIs masitinib and toceranib might be considered the first-line therapy for unresectable and/or metastatic canine MCT, but also for those cases with confirmed internal tandem duplications in the exon 11 of the c-KIT protooncogene. Masitinib appears to be more selective than others TKI, such as toceranib, imatinib, dasatinib and sunitinib, because it causes weak inhibition of BCR/ABL (breakpoint cluster region-Abelson), Fms (macrophage colony-stimulating factor receptor), Flt-3 (FMS-like tyrosine kinase-3) and VEGFR (vascular endothelial growth factor receptor), which may partially explains its increased safety and lower risk of cardiotoxicity. In the first case, the animal has been treated with lomustine associated to masitinib and showed a progression-free interval of 33 days, however, the response reported may have been lower, due previously exposition to chemotherapeutic agents, which might compromise the response to TKI. The second case, with the association of lomustine and toceranib, was followed up for 365 days, presenting only one recurrence in the final third of the follow-up, however, with subsequent new complete remission. Sensitization of canine MCT to lomustine with TKIs increases the therapeutic possibilities for this neoplasm, mainly in patients with advanced stage and high-grade tumours.

The influence of season, environment, sex, and body mass on biochemical profile of the freshwater turtle Phrynops geoffroanus (Schweigger, 1812) / [Influência da estação, do ambiente, do sexo e da massa corporal no perfil bioquímico do cágado Phrynops geoffroanus (Schweigger, 1812)]

We analyzed the influence of the season, the environment, and the sex, as well as the relation of body mass (BM) in the serum albumin (ALB), aspartate aminotransferase (AST), creatinine (C), creatine kinase (CK), phosphorus (P), total calcium (tCa), total protein (TP), urea (U), uric acid (UA), calcium:phosphorus ratio (Ca:P), and the globulin value (GV) of thirty individuals of Phrynops geoffroanus of the urban area of Cuiabá, Mato Grosso, Brazil. The modeling of biochemical parameters was performed using the Generalized Additive Models for Location, Scale and Shape (GAMLSS) to verify the influence of variables considered in this study on each of the biochemical parameters analyzed. The season influenced AST, CK, C, tCa, Ca:P and UA. The environment influenced tCa, Ca:P, U and UA. On the other hand, CK, tCa, P, Ca:P and U differed significantly between males and females. Regarding the BM, a relationship of this variable was observed with CK, C, tCa, P, U, UA and Ca:P. We concluded that the season, environment, sex, and body mass can influence the biochemical parameters of P. geoffroanus, and these factors should be routinely considered in the interpretation of laboratory results.

Analisou-se a influência da estação, do ambiente e do sexo, assim como a relação da massa corporal (BM) nos níveis sorológicos de albumina (ALB), aspartato aminotransferase (AST), creatinina (C), creatina quinase (CK), fósforo (P), cálcio total (tCa), sólidos totais (TS), ureia (U), ácido úrico (UA), relação cálcio:fósforo (Ca:P), e do valor da globulina (GV) de Phrynops geoffroanus da área urbana de Cuiabá, Mato Grosso, Brasil. A modelagem dos parâmetros bioquímicos foi realizada utilizando-se os modelos aditivos generalizados para locação, escala e forma (GAMLSS) para verificar a influência das variáveis consideradas neste estudo em cada um dos parâmetros bioquímicos analisados. A estação sazonal influenciou os níveis de AST, CK, C, tCa, Ca:P e UA. O ambiente foi capaz de influenciar tCa, Ca:P, U e UA. Por outro lado, CK, tCa, P, Ca:P e U diferiram significativamente entre machos e fêmeas. Em relação à BM, observou-se relação dessa variável com CK, C, tCa, P, U, UA e Ca:P. Concluiu-se que a estação sazonal, o ambiente, o sexo e a massa corporal são capazes de influenciar os parâmetros bioquímicos de P. geoffroanus e que esses fatores devem ser rotineiramente considerados na interpretação dos resultados laboratoriais.

The neuroprotection of cerebrolysin after spontaneous intracerebral hemorrhage through regulates necroptosis via Akt/ GSK3B signaling pathway

ABSTRACT Purpose: Spontaneous intracerebral hemorrhage (ICH) is a major cause of death and disability with a huge economic burden worldwide. Cerebrolysin (CBL) has been previously used as a nootropic drug. Necroptosis is a programmed cell death mechanism that plays a vital role in neuronal cell death after ICH. However, the precise role of necroptosis in CBL neuroprotection following ICH has not been confirmed. Methods: In the present study, we aimed to investigate the neuroprotective effects and potential molecular mechanisms of CBL in ICH-induced early brain injury (EBI) by regulating neural necroptosis in the C57BL/6 mice model. Mortality, neurological score, brain water content, and neuronal death were evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, Evans blue extravasation, Western blotting, and quantitative real-time polymerase chain reaction (PCR). Results: The results show that CBL treatment markedly increased the survival rate, neurological score, and neuron survival, and downregulated the protein expression of RIP1 and RIP3, which indicated that CBL-mediated inhibition of necroptosis, and ameliorated neuronal death after ICH. The neuroprotective capacity of CBL is partly dependent on the Akt/GSK3β signaling pathway. Conclusions: CBL improves neurological outcomes in mice and reduces neuronal death by protecting against neural necroptosis.

Effects of different stimulators of cGMP synthesis on lipid content in bovine oocytes matured in vitro

Bovine oocytes and blastocysts produced in vitro are frequently of lower quality and less cryotolerant than those produced in vivo, and greater accumulation of lipids in the cytoplasm has been pointed out as one of the reasons. In human adipocytes cGMP signaling through the activation of PKG appears to be involved in lipid metabolism, and components of this pathway have been detected in bovine cumulus-oocyte complexes (COCs). The aim of this study was to investigate the influence of this pathway on the lipid content in oocytes and expression of PLIN2 (a lipid metabolism-related gene) in cumulus cells. COCs were matured in vitro for 24 h with different stimulators of cGMP synthesis. The activation of soluble guanylyl cyclase (sGC) by Protoporphyrin IX reduced lipid content (22.7 FI) compared to control oocytes (36.45 FI; P <0.05). Stimulation of membrane guanylyl cyclase (mGC) with natriuretic peptides precursors A and C (NPPA and NPPC) had no effect (36.5 FI; P>0.05). When the PKG inhibitor KT5823 was associated with Protoporphyrin IX, its effect was reversed and lipid contents increased (52.71 FI; P<0.05). None of the stimulators of cGMP synthesis affected the expression of PLIN2 in cumulus cells. In conclusion, stimulation of sGC for cGMP synthesis promotes lipolytic activities in bovine oocytes matured in vitro and such effect is mediated by PKG. However, such effect may vary depending on the stimulus received and/or which synthesis enzyme was activated, as stimulation of mGC had no effects.(AU)

Use of protein kinase C and phospholipase A2 inhibitors in bovine endometrial cells treated with estradiol and calcium ionophore / Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio

The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 µM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 µM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17ß-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2.(AU)

A liberação endometrial de prostaglandina-F2α (PGF2α) em fêmeas bovinas pode ser induzida in vivo pelo estradiol (E2). Entretanto o seu mecanismo de ação ainda não foi bem esclarecido. Nossa hipótese é que o E2 estimula a atividade e a abundância da proteína quinase C (PKC) e da fosfolipase A2 (PLA2). Nosso objetivo com este estudo foi analizar os efeitos de inibidores de PKC e PLA2 na síntese de PGF2α induzida por E2 e ionóforo de cálcio (CI) em células endometriais bovinas (células BEND; Experimento 1). Adicionalmente, nós avaliamos a abundância de PKC e PLA2 em explantes endometriais de vacas tratadas com ou sem E2 17 dias após o estro (D17, D0 = estro; Experimento 2). No Experimento 1, células BEND foram submetidas ao inibidor de PKC (10 µM de C25H24N4O2; bisindolylmaleimide I, ou BIS I), e ao inibidor de PLA2 (20 µM de arachydoniltrifluoromethane ou AACOCF3) ou a nenhum inibidor. As células BEND foram subsequentemente tratadas com E2 e CI e concentrações de PGF2α foram mensuradas no meio de cultura por radioimunoenssaio. Para DIF-12 (concentração de PGF2α 12 horas depois do tratamento, subtraída da concentração de PGF2α na hora 0), não foi observado efeito do inibidor de PKC (P = 0.2709). Entretanto DIF-12 foi menor (P < 0.05) nos grupos tratados com inibidor de PLA2 e inibidor de PLA2 + CI + E2 quando comparados com o grupo controle e o grupo CI + E2. O AACOCF3 foi um eficiente inibidor de PLA2 em sistema de cultura de células BEND e o E2 não estimulou a síntese de PKC e PLA2. No Experimento 2, novilhas Nelore cíclicas receberam 3 mg de 17ß-E2 (n = 6) ou nenhum tratamento (n = 5) no D17 e foram abatidas duas horas depois da administração dos tratamentos. A quantidade de PKC and PLA2 no tecido endometrial foi avaliada pela técnica de Western Blotting. Não foi observado efeito do E2 sobre a PKC (P= 0.08) e nem sobre a PLA2 (P= 0.56). Conclui-se que o E2 não estimulou a atividade e abundância de PKC e PLA2.(AU)

Use of protein kinase C and phospholipase A2 inhibitors in bovine endometrial cells treated with estradiol and calcium ionophore / Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio

The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 µM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 µM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17ß-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2.(AU)

A liberação endometrial de prostaglandina-F2α (PGF2α) em fêmeas bovinas pode ser induzida in vivo pelo estradiol (E2). Entretanto o seu mecanismo de ação ainda não foi bem esclarecido. Nossa hipótese é que o E2 estimula a atividade e a abundância da proteína quinase C (PKC) e da fosfolipase A2 (PLA2). Nosso objetivo com este estudo foi analizar os efeitos de inibidores de PKC e PLA2 na síntese de PGF2α induzida por E2 e ionóforo de cálcio (CI) em células endometriais bovinas (células BEND; Experimento 1). Adicionalmente, nós avaliamos a abundância de PKC e PLA2 em explantes endometriais de vacas tratadas com ou sem E2 17 dias após o estro (D17, D0 = estro; Experimento 2). No Experimento 1, células BEND foram submetidas ao inibidor de PKC (10 µM de C25H24N4O2; bisindolylmaleimide I, ou BIS I), e ao inibidor de PLA2 (20 µM de arachydoniltrifluoromethane ou AACOCF3) ou a nenhum inibidor. As células BEND foram subsequentemente tratadas com E2 e CI e concentrações de PGF2α foram mensuradas no meio de cultura por radioimunoenssaio. Para DIF-12 (concentração de PGF2α 12 horas depois do tratamento, subtraída da concentração de PGF2α na hora 0), não foi observado efeito do inibidor de PKC (P = 0.2709). Entretanto DIF-12 foi menor (P < 0.05) nos grupos tratados com inibidor de PLA2 e inibidor de PLA2 + CI + E2 quando comparados com o grupo controle e o grupo CI + E2. O AACOCF3 foi um eficiente inibidor de PLA2 em sistema de cultura de células BEND e o E2 não estimulou a síntese de PKC e PLA2. No Experimento 2, novilhas Nelore cíclicas receberam 3 mg de 17ß-E2 (n = 6) ou nenhum tratamento (n = 5) no D17 e foram abatidas duas horas depois da administração dos tratamentos. A quantidade de PKC and PLA2 no tecido endometrial foi avaliada pela técnica de Western Blotting. Não foi observado efeito do E2 sobre a PKC (P= 0.08) e nem sobre a PLA2 (P= 0.56). Conclui-se que o E2 não estimulou a atividade e abundância de PKC e PLA2.(AU)

Involvement of HSP90 in ischemic postconditioning-induced cardioprotection by inhibition of the complement system, JNK and inflammation

Purpose To investigate whether heat shock protein 90 (HSP90) is involved in complement regulation in ischemic postconditioning (IPC). Methods The left coronary artery of rats underwent 30 min of occlusion, followed by 120 min of reperfusion and treatment with IPC via 3 cycles of 30s reperfusion and 30s occlusion. The rats were injected intraperitoneally with 1 mg/kg HSP90 inhibitor geldanamycin (GA) after anesthesia. Eighty rats were randomly divided into four groups: sham, ischemia-reperfusion (I/R), IPC and IPC + GA. Myocardial infarct size, apoptosis index and the expression of HSP90, C3, C5a, tumor necrosis factor (TNF)-alpha, interleukin (IL)-1β and c-Jun N-terminal kinase (JNK) were assessed. Results Compared with the I/R injury, the IPC treatment significantly reduced infarct size, release of troponin T, creatine kinase-MB, and lactate dehydrogenase, and cardiomyocyte apoptosis. These beneficial effects were accompanied by a decrease in TNF-α, IL-1β, C3, C5a and JNK expression levels. However, all these effects were abrogated by administration of the HSP90 inhibitor GA. Conclusion HSP90 exerts a profound effect on IPC cardioprotection, and may be linked to the inhibition of the complement system and JNK, ultimately attenuating I/R-induced myocardial injury and apoptosis.(AU)

Protective effect of L-carnitine on myocardial injury in rats with heatstroke

Purpose: To investigate the protective effect of L-carnitine on myocardial injury in rats with heatstroke. Methods: orty-eight rats were randomly divided into control, heatstroke and 25, 50 and 100 mg/kg L-carnitine groups. The last three groups were treated with 25, 50 and 100 mg/kg L-carnitine, respectively, for seven successive days. Then, except for the control group, the other four groups were transferred into the environment with ambient temperature of (39.5 ± 0.4 °C) and relative humidity of (13.5 ± 2.1%) for 2 h. The core temperature (Tc), mean arterial pressure (MAP), heart rate (HR) and serum and myocardial indexes were detected. Results: Compared with the heatstroke group, in the 100 mg/kg L-carnitine group, the Tc was significantly decreased, the MAP and HR were significantly increased, the serum creatine kinase, lactate dehydrogenase, alkaline phosphatase, aspartate aminotransferase, tumor necrosis factor and interleukin 1 levels were significantly decreased, the myocardial superoxide dismutase and glutathione peroxidase levels were significantly increased, the myocardial malondialdehyde level was significantly decreased and the cardiomyocyte apoptosis index and myocardial caspase-3 protein expression level were remarkably decreased (p 0.05). Conclusions: The L-carnitine pretreatment can alleviate the myocardial injury in heatstroke rats through reducing the inflammatory response, oxidative stress and cardiomyocyte apoptosis.(AU)

BjussuLAAO-II induces cytotoxicity and alters DNA methylation of cell-cycle genes in monocultured/co-cultured HepG2 cells

Background:The use of animal venoms and their toxins as material sources for biotechnological applications has received much attention from the pharmaceutical industry. L-amino acid oxidases from snake venoms (SV-LAAOs) have demonstrated innumerous biological effects and pharmacological potential against different cancer types. Hepatocellular carcinoma has increased worldwide, and the aberrant DNA methylation of liver cells is a common mechanism to promote hepatic tumorigenesis. Moreover, tumor microenvironment plays a major role in neoplastic transformation. To elucidate the molecular mechanisms responsible for the cytotoxic effects of SV-LAAO in human cancer cells, this study aimed to evaluate the cytotoxicity and the alterations in DNA methylation profiler in the promoter regions of cell-cycle genes induced by BjussuLAAO-II, an LAAO from Bothrops jaracussu venom, in human hepatocellular carcinoma (HepG2) cells in monoculture and co-culture with endothelial (HUVEC) cells.Methods:BjussuLAAO-II concentrations were 0.25, 0.50, 1.00 and 5.00 μg/mL. Cell viability was assessed by MTT assay and DNA methylation of the promoter regions of 22 cell-cycle genes by EpiTect Methyl II PCR array.Results:BjussuLAAO-II decreased the cell viability of HepG2 cells in monoculture at all concentrations tested. In co-culture, 1.00 and 5.00 μg/mL induced cytotoxicity (p < 0.05). BjussuLAAO-II increased the methylation of CCND1 and decreased the methylation of CDKN1A in monoculture and GADD45A in both cell-culture models (p < 0.05).Conclusion:Data showed BjussuLAAO-II induced cytotoxicity and altered DNA methylation of the promoter regions of cell-cycle genes in HepG2 cells in monoculture and co-culture models. We suggested the analysis of DNA methylation profile of GADD45A as a potential biomarker of the cell cycle effects of BjussuLAAO-II in cancer cells...(AU)

Avaliação da atividade da proteína quinase P34Cdc2 na maturação in vitro de oócitos caninos / Evaluation of p34cdc2 kinase activity in the in vitro maturation of canine oocytes

O controle do ciclo celular é regulado por uma cascata de eventos coordenados que podem atuar influenciando na expressão ou repressão da atividade de proteínas relacionadas a retomada da meiose. Estudos indicam que a atividade dessas proteínas mostra-se tempo-dependente no processo de maturação in vitro (MIV). Esse trabalho teve o objetivo de avaliar a cinética da atividade quinase p34cdc2 durante a MIV de oócitos caninos. Ciência Animal 27(1), 2017. Os ovários foram obtidos de 40 cadelas submetidas à ovário-salpingo-histerectomia (OSH) eletiva. Após a OSH, os ovários foram imediatamente transportados a uma temperatura de 4 ºC. No laboratório, os ovários foram seccionados em fatias finas ("slicing"), para a liberação dos complexos cumulus-oócito (COCs). Apenas os COCs grau 1 foram selecionados e colocados em meio de maturação por um período de 24, 48 e 72 h de maturação. Após o cultivo, os COCs foram colocados em placas contendo solução de hialuronidase 0.2% para a retirada completa das células do Cumulus. A atividade da proteína p34cdc2 foi detectada por ELISA. Com base nos resultados verifica-se que a atividade da proteína mostra-se tempo-dependente, atingindo o pico após 48h de MIV (p<0,01). Após 72h, a atividade demonstrou um decréscimo. Com base neste estudo pode-se concluir que a proteína quinase p34cdc2 desempenha uma função de suma importância na progressão da meiose em cadelas. Dessa forma, a compreensão melhor dessa proteína assim como de outra que participam do processo de maturação poderá contribuir para o estabelecimento de meios mais adequados que melhorem significativamente as taxas dematuração.(AU)

The control of the cell cycle is regulated by a cascade of coordinated events that can act by influencing the expression or repression of the activity of proteins related to meiosis resumption. Studies have indicated that the activity of these proteins is time-dependent in the in vitro maturation process (IVM). This work aimed to evaluate the kinetics of the p34cdc2 kinase activity during IVM of canine oocytes. Ovaries were obtained from 40 bitches submitted to elective ovary-salpingo-hysterectomy (OSH). After OSH, ovaries were immediately transported at temperature of 4 °C. In the laboratory, ovaries were sliced for the release of cumulus-oocyte complexes (COCs). Only grade 1 COCs were selected and placed in maturation medium for a period of 24, 48 and 72 h of maturation. After culturing, COCs were plated of 0.2% hyaluronidase solution for complete removal of cumulus cells. The activity of the p34cdc2 protein was detected by ELISA. Based on the results, it was observed that the activity of the protein is time-dependent, peaking after 48 hours of IVM (p 0.01). After 72 hours, activity declined. Based on this study, it could be concluded that the p34cdc2 protein kinase plays a very important role in the meiosis progression in bitches. Thus, a better understanding of this protein as well as of others that participate in the maturation process may contribute to the establishment of more adequate media to significantly improve maturation rates.(AU)

Expression of EGF receptors in canine prostate with proliferative inflammatory atrophy and carcinoma / Expressão de receptores EGF na próstata canina com atrofia inflamatória proliferativa e carcinoma

Gene expression of ErbB1 and ErbB2, and immunostaining of EGFR (Her1) and Her2 (c-erbB-2) were evaluated in this study to ascertain whether these receptors are involved in the evolution of canine premalignant and malignant prostatic lesions, as proliferative inflammatory atrophy (PIA) and prostatic carcinoma (PC). With regards to the intensity of EGFR immunostaining, there was no difference between normal prostatic tissue and tissues with PIA or PC. In relation to Her2 immunostaining, there were differences between normal prostatic tissue and those with PIA and PC, as also differences between prostates with PIA and PC. There was no correlation between EGFR and Her2 immunostaining. ErbB1 gene product was detected in two normal tissue samples, in one with PIA, and in all samples with PC. ErbB2 mRNA was recorded in two canine samples with PIA, in all with PC, but was not detected in normal prostatic tissue. It was concluded that EGFR and Her2 play roles in canine PIA and PC, suggesting that those receptors may be involved in canine prostatic carcinogenesis.

A expressão gênica de ErbB1 e ErbB2 e a imunomarcação de EGFR (Her1) e Her2 (c-erbB-2) foram avaliadas para verificar o envolvimento desses receptores em lesões pré-malignas e malignas da próstata canina, como a atrofia proliferativa inflamatória (PIA) e o carcinoma prostático (PC). Em relação à intensidade de imunomarcação para EGFR, não houve diferença entre o tecido prostático normal e com PIA e PC. Em relação a Her2, observou-se diferença de imunomarcação entre o tecido prostático normal e aqueles com PIA e PC e entre os com PIA e PC. Não houve correlação entre EGFR e Her2. O gene ErbB1 foi detectado em duas amostras normais, uma de PIA e em todas as amostras de PC. O gene ErbB2 foi detectado em duas amostras de PIA e em todas as amostras de PC, não sendo detectado no tecido prostático normal. Conclui-se que EGFR e Her2 atuam nas lesões de PIA e PC, sugerindo o envolvimento destes na carcinogênese da próstata canina.

Expression of EGF receptors in canine prostate with proliferative inflammatory atrophy and carcinoma / Expressão de receptores EGF na próstata canina com atrofia inflamatória proliferativa e carcinoma

Gene expression of ErbB1 and ErbB2, and immunostaining of EGFR (Her1) and Her2 (c-erbB-2) were evaluated in this study to ascertain whether these receptors are involved in the evolution of canine premalignant and malignant prostatic lesions, as proliferative inflammatory atrophy (PIA) and prostatic carcinoma (PC). With regards to the intensity of EGFR immunostaining, there was no difference between normal prostatic tissue and tissues with PIA or PC. In relation to Her2 immunostaining, there were differences between normal prostatic tissue and those with PIA and PC, as also differences between prostates with PIA and PC. There was no correlation between EGFR and Her2 immunostaining. ErbB1 gene product was detected in two normal tissue samples, in one with PIA, and in all samples with PC. ErbB2 mRNA was recorded in two canine samples with PIA, in all with PC, but was not detected in normal prostatic tissue. It was concluded that EGFR and Her2 play roles in canine PIA and PC, suggesting that those receptors may be involved in canine prostatic carcinogenesis.(AU)

A expressão gênica de ErbB1 e ErbB2 e a imunomarcação de EGFR (Her1) e Her2 (c-erbB-2) foram avaliadas para verificar o envolvimento desses receptores em lesões pré-malignas e malignas da próstata canina, como a atrofia proliferativa inflamatória (PIA) e o carcinoma prostático (PC). Em relação à intensidade de imunomarcação para EGFR, não houve diferença entre o tecido prostático normal e com PIA e PC. Em relação a Her2, observou-se diferença de imunomarcação entre o tecido prostático normal e aqueles com PIA e PC e entre os com PIA e PC. Não houve correlação entre EGFR e Her2. O gene ErbB1 foi detectado em duas amostras normais, uma de PIA e em todas as amostras de PC. O gene ErbB2 foi detectado em duas amostras de PIA e em todas as amostras de PC, não sendo detectado no tecido prostático normal. Conclui-se que EGFR e Her2 atuam nas lesões de PIA e PC, sugerindo o envolvimento destes na carcinogênese da próstata canina.(AU)

Prognostic factors in canine mammary carcinomas and HER2 expression relationship / Fatores de prognóstico em carcinomas mamários caninos e sua relação com expressão de HER2

Background: The human epidermal growth factor type 2 (HER2) receptor is a membrane glycoprotein tyrosine kinase. In woman, HER2 expression is diagnosed in 30% of breast carcinomas and it is associated with a worse prognosis, higher rate of recurrence and mortality. In the bitch, the HER2 overexpression in canine mammary tumors is still controversial and the prognostic value remains uncertain. Thus, we aimed to verify the HER2 expression in canine mammary carcinomas and relate it to the type and histological grade, lymph node metastasis and clinical staging. Materials, Methods & Results: Ninety bitches diagnosed with mammary carcinoma were included in this study. The inclusion criteria were bitches with complete clinical examination, thoracic radiographic examination and submitted unilateral or bilateral mastectomy. Ninety-nine samples of mammary carcinoma were used and the fragments of tumor and regional lymph nodes were fixed in 10% neutral formalin for histopathological and immunohistochemistry analysis. The lesions were evaluated by two pathologists and classified according to the type and histological grade. HER2 expression was performed by semi-quantitative analysis of the slides according to the HerceptTestTM (Dako) recommended score. Simple carcinomas were the most frequent (51.51%) followed by complex carcinomas (46.47%) and in situ carcinoma (2.02%). [...](AU)

Prognostic factors in canine mammary carcinomas and HER2 expression relationship / Fatores de prognóstico em carcinomas mamários caninos e sua relação com expressão de HER2

Background: The human epidermal growth factor type 2 (HER2) receptor is a membrane glycoprotein tyrosine kinase. In woman, HER2 expression is diagnosed in 30% of breast carcinomas and it is associated with a worse prognosis, higher rate of recurrence and mortality. In the bitch, the HER2 overexpression in canine mammary tumors is still controversial and the prognostic value remains uncertain. Thus, we aimed to verify the HER2 expression in canine mammary carcinomas and relate it to the type and histological grade, lymph node metastasis and clinical staging. Materials, Methods & Results: Ninety bitches diagnosed with mammary carcinoma were included in this study. The inclusion criteria were bitches with complete clinical examination, thoracic radiographic examination and submitted unilateral or bilateral mastectomy. Ninety-nine samples of mammary carcinoma were used and the fragments of tumor and regional lymph nodes were fixed in 10% neutral formalin for histopathological and immunohistochemistry analysis. The lesions were evaluated by two pathologists and classified according to the type and histological grade. HER2 expression was performed by semi-quantitative analysis of the slides according to the HerceptTestTM (Dako) recommended score. Simple carcinomas were the most frequent (51.51%) followed by complex carcinomas (46.47%) and in situ carcinoma (2.02%). [...]

Avaliação da atividade da proteína quinase P34Cdc2 na maturação in vitro de oócitos caninos / Evaluation of p34cdc2 kinase activity in the in vitro maturation of canine oocytes

O controle do ciclo celular é regulado por uma cascata de eventos coordenados que podem atuar influenciando na expressão ou repressão da atividade de proteínas relacionadas a retomada da meiose. Estudos indicam que a atividade dessas proteínas mostra-se tempo-dependente no processo de maturação in vitro (MIV). Esse trabalho teve o objetivo de avaliar a cinética da atividade quinase p34cdc2 durante a MIV de oócitos caninos. Ciência Animal 27(1), 2017. Os ovários foram obtidos de 40 cadelas submetidas à ovário-salpingo-histerectomia (OSH) eletiva. Após a OSH, os ovários foram imediatamente transportados a uma temperatura de 4 ºC. No laboratório, os ovários foram seccionados em fatias finas ("slicing"), para a liberação dos complexos cumulus-oócito (COCs). Apenas os COCs grau 1 foram selecionados e colocados em meio de maturação por um período de 24, 48 e 72 h de maturação. Após o cultivo, os COCs foram colocados em placas contendo solução de hialuronidase 0.2% para a retirada completa das células do Cumulus. A atividade da proteína p34cdc2 foi detectada por ELISA. Com base nos resultados verifica-se que a atividade da proteína mostra-se tempo-dependente, atingindo o pico após 48h de MIV (p<0,01). Após 72h, a atividade demonstrou um decréscimo. Com base neste estudo pode-se concluir que a proteína quinase p34cdc2 desempenha uma função de suma importância na progressão da meiose em cadelas. Dessa forma, a compreensão melhor dessa proteína assim como de outra que participam do processo de maturação poderá contribuir para o estabelecimento de meios mais adequados que melhorem significativamente as taxas dematuração.

The control of the cell cycle is regulated by a cascade of coordinated events that can act by influencing the expression or repression of the activity of proteins related to meiosis resumption. Studies have indicated that the activity of these proteins is time-dependent in the in vitro maturation process (IVM). This work aimed to evaluate the kinetics of the p34cdc2 kinase activity during IVM of canine oocytes. Ovaries were obtained from 40 bitches submitted to elective ovary-salpingo-hysterectomy (OSH). After OSH, ovaries were immediately transported at temperature of 4 °C. In the laboratory, ovaries were sliced for the release of cumulus-oocyte complexes (COCs). Only grade 1 COCs were selected and placed in maturation medium for a period of 24, 48 and 72 h of maturation. After culturing, COCs were plated of 0.2% hyaluronidase solution for complete removal of cumulus cells. The activity of the p34cdc2 protein was detected by ELISA. Based on the results, it was observed that the activity of the protein is time-dependent, peaking after 48 hours of IVM (p 0.01). After 72 hours, activity declined. Based on this study, it could be concluded that the p34cdc2 protein kinase plays a very important role in the meiosis progression in bitches. Thus, a better understanding of this protein as well as of others that participate in the maturation process may contribute to the establishment of more adequate media to significantly improve maturation rates.

Insulin binding characteristics in canine muscle tissue: effects of the estrous cycle phases / Características de ligação da insulina no tecido muscular canino: efeitos da fase do ciclo estral

Hormonal fluctuations during the different estrous cycle are a well-recognized cause of insulin resistance in bitches, and little is known about insulin receptor binding or post-binding defects associated with insulin resistance in dogs. To evaluate insulin binding characteristics in muscle tissue of bitches during the estrous cycle, 17 owned bitches were used in the study (six in anestrus, five in estrus, and six in diestrus). An intravenous glucose tolerance test (IVGTT) was performed in all patients by means of injection of 1mL/kg of a glucose 50% solution (500mg/kg), with blood sample collection for glucose determination at 0, 3, 5, 7, 15, 30, 45 and 60 minutes after glucose infusion. Muscle samples, taken after spaying surgery, were immediately frozen in liquid nitrogen and then stored at -80 ºC until the membranes were prepared by sequential centrifugation after being homogenized. For binding studies, membranes were incubated in the presence of 20,000cpm of human 125I-insulin and in increasing concentrations of unlabeled human regular insulin for cold saturation. The IVGTT showed no differences among bitches during the estrous cycle regarding baseline glycemia or glycemic response after glucose infusion. Two insulin binding sites - high-affinity and low-affinity ones - were detected by Scatchard analysis, and significant statistical differences were observed in the dissociation constant (Kd1) and maximum binding capacity (Bmax1) of the high-affinity binding sites. The Kd1 for the anestrus group (6.54±2.77nM/mg of protein) was smaller (P<0.001) than for the estrus (28.54±6.94nM/mg of protein) and diestrus (15.56±3.88nM/mg of protein) groups. Bmax1 in the estrus (0.83±0.42nM/mg of protein) and diestrus (1.24±0.24nM/mg of protein) groups were also higher (P<0.001) than the values observed in anestrus (0.35±0.06nM/mg of protein). These results indicate modulation of insulin binding characteristics during different phases of the estrous cycle in dogs, showing that muscle insulin binding affinity for its receptor is reduced during estrus and diestrus. However, this poor hormone-receptor affinity is compensated for by a greater total binding capacity, once there is no difference in patients' glycemic response after an intravenous glucose load.(AU)

As flutuações hormonais durante as diferentes fases do ciclo estral são uma causa importante de resistência insulínica em fêmeas caninas, e poucas informações são conhecidas sobre defeitos na ligação da insulina ao seu receptor, ou defeitos pós-receptor associados com resistência à insulina em cães. Para avaliar as características da ligação insulina-receptor no tecido muscular de cadelas durante o ciclo estral, dezessete pacientes foram utilizadas no estudo (seis em anestro, cinco em estro e seis em diestro). Um teste de tolerância à glicose intravenosa (IVGTT) foi realizado em todas as pacientes por meio da infusão de 1mL/kg de uma solução de glicose 50% (500mg/kg), com coletas de sangue para determinação de glicemia nos tempos 0, 3, 5, 7, 15, 30, 45 e 60 minutos da injeção de glicose. Amostras de tecido muscular foram coletadas durante ovariohisterectomia, imediatamente congeladas em nitrogênio líquido, e posteriormente armazenadas a -80°C até a preparação das membranas por meio de homogeneização e centrifugação sequencial. Para os experimentos de ligação hormônio-receptor, as membranas foram incubadas na presença de 20.000cpm de 125I-insulina humana, e concentrações crescentes de insulina regular humana não marcada para saturação fria. O IVGTT não mostrou diferenças entre as pacientes em diferentes fases do ciclo estral com relação a glicemia basal, ou na resposta glicêmica após infusão de glicose nos tempos estudados. Dois sítios de ligação da insulina, um de alta-afinidade, e outro de baixa afinidade, foram detectados pela análise de Scatchard, e diferenças significativas foram detectadas na constante de dissociação (Kd1) e capacidade de ligação máxima (Bmax1) dos sítios de ligação de alta-afinidade. O Kd1 para o grupo anestro (6,54±2,77nM/mg de proteína) foi menor (P<0,001) que os Kd1 dos grupos estro (28,54±6,94 nM/mg de proteína) e diestro (15,56±3,88nM/mg de proteína). Os Bmax1 dos grupos estro (0,83±0,42nM/mg de proteína) e diestro (1,24±0,24nM/mg de proteína) também foram maiores que os valores encontrados no grupo anestro (0,35±0,06nM/mg de proteína). Estes resultados demonstram uma modulação das características de ligação da insulina nas diferentes fases do ciclo estral em cães, evidenciando uma menor afinidade de ligação da insulina ao seu receptor no tecido muscular durante o estro e diestro. Contudo, esta menor afinidade de ligação hormônio-receptor é compensada por uma maior capacidade de ligação, o que fica também evidenciado pela ausência de diferenças na resposta glicêmica das pacientes após um desafio com glicose por via endovenosa.(AU)

Insulin binding characteristics in canine muscle tissue: effects of the estrous cycle phases / Características de ligação da insulina no tecido muscular canino: efeitos da fase do ciclo estral

Hormonal fluctuations during the different estrous cycle are a well-recognized cause of insulin resistance in bitches, and little is known about insulin receptor binding or post-binding defects associated with insulin resistance in dogs. To evaluate insulin binding characteristics in muscle tissue of bitches during the estrous cycle, 17 owned bitches were used in the study (six in anestrus, five in estrus, and six in diestrus). An intravenous glucose tolerance test (IVGTT) was performed in all patients by means of injection of 1mL/kg of a glucose 50% solution (500mg/kg), with blood sample collection for glucose determination at 0, 3, 5, 7, 15, 30, 45 and 60 minutes after glucose infusion. Muscle samples, taken after spaying surgery, were immediately frozen in liquid nitrogen and then stored at -80 ºC until the membranes were prepared by sequential centrifugation after being homogenized. For binding studies, membranes were incubated in the presence of 20,000cpm of human 125I-insulin and in increasing concentrations of unlabeled human regular insulin for cold saturation. The IVGTT showed no differences among bitches during the estrous cycle regarding baseline glycemia or glycemic response after glucose infusion. Two insulin binding sites - high-affinity and low-affinity ones - were detected by Scatchard analysis, and significant statistical differences were observed in the dissociation constant (Kd1) and maximum binding capacity (Bmax1) of the high-affinity binding sites. The Kd1 for the anestrus group (6.54±2.77nM/mg of protein) was smaller (P<0.001) than for the estrus (28.54±6.94nM/mg of protein) and diestrus (15.56±3.88nM/mg of protein) groups. Bmax1 in the estrus (0.83±0.42nM/mg of protein) and diestrus (1.24±0.24nM/mg of protein) groups were also higher (P<0.001) than the values observed in anestrus (0.35±0.06nM/mg of protein). [...] (AU)

As flutuações hormonais durante as diferentes fases do ciclo estral são uma causa importante de resistência insulínica em fêmeas caninas, e poucas informações são conhecidas sobre defeitos na ligação da insulina ao seu receptor, ou defeitos pós-receptor associados com resistência à insulina em cães. Para avaliar as características da ligação insulina-receptor no tecido muscular de cadelas durante o ciclo estral, dezessete pacientes foram utilizadas no estudo (seis em anestro, cinco em estro e seis em diestro). Um teste de tolerância à glicose intravenosa (IVGTT) foi realizado em todas as pacientes por meio da infusão de 1mL/kg de uma solução de glicose 50% (500mg/kg), com coletas de sangue para determinação de glicemia nos tempos 0, 3, 5, 7, 15, 30, 45 e 60 minutos da injeção de glicose. Amostras de tecido muscular foram coletadas durante ovariohisterectomia, imediatamente congeladas em nitrogênio líquido, e posteriormente armazenadas a -80°C até a preparação das membranas por meio de homogeneização e centrifugação sequencial. Para os experimentos de ligação hormônio-receptor, as membranas foram incubadas na presença de 20.000cpm de 125I-insulina humana, e concentrações crescentes de insulina regular humana não marcada para saturação fria. O IVGTT não mostrou diferenças entre as pacientes em diferentes fases do ciclo estral com relação a glicemia basal, ou na resposta glicêmica após infusão de glicose nos tempos estudados. Dois sítios de ligação da insulina, um de alta-afinidade, e outro de baixa afinidade, foram detectados pela análise de Scatchard, e diferenças significativas foram detectadas na constante de dissociação (Kd1) e capacidade de ligação máxima (Bmax1) dos sítios de ligação de alta-afinidade. O Kd1 para o grupo anestro (6,54±2,77nM/mg de proteína) foi menor (P<0,001) que os Kd1 dos grupos estro (28,54±6,94 nM/mg de proteína) e diestro (15,56±3,88nM/mg de proteína).[...] (AU)

Serum hepatic biochemistry and electrophoretic protein profile of healthy and Ehrlich tumor-bearing mice treated with extracts of Agaricus blazei Murill / Bioquímica hepática sérica e perfil proteico eletroforético de camundongos saudáveis ou portadores do tumor de Ehrlich tratados com frações de extratos de Agaricus blazei Murril

Compounds isolated from Agaricus blazei Murill represent a group of promising natural immunomodulators for use in the treatment of neoplasms. We have evaluated the serum biochemical profile of healthy and Ehrlich tumor-bearing mice treated with different extracts of A. blazei. Total, supernatant, and polysaccharide extracts of A. blazei were obtained from suspensions (at acidic or neutral pH) kept in a water bath at 60 °C or in an ultrasonic bath at 37 °C. After oral administering the extracts to mice for 21 days, blood samples were collected for determination of aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), urea, total protein, albumin, globulins, and alpha-, beta- and gamma-globulin fractions. The presence of the tumor led to a significant increase in serum CK and AST activities and in the concentrations of total globulin and the gamma-globulin fraction, and to a decrease in the albumin and alpha2-globulin levels. The polysaccharide extracts of A. blazei reduced the serum AST and ALT activities, probably due to a hepatoprotective effect. In addition, polysaccharide and supernatant extracts inhibited the tumor-induced increase in gamma-globulin levels. Thus, the supernatant and polysaccharide fractions of the extract of A. blazei have potential for use in complementary antineoplastic treatments.(AU)

Biocompostos de Agaricus blazei Murril representam imunomoduladores naturais promissores para uso no tratamento de neoplasias. Objetivou-se avaliar o perfil bioquímico hepático sérico camundongos saudáveis ou portadores do tumor de Ehrlich tratados com diferentes extratos de A. blazei. Em pH ácido ou neutro e sob temperatura de extração de 60 ºC em banho-maria ou 37 ºC em banho ultrassônico foram obtidos extratos totais, de sobrenadante e de polissacarídeos de A. blazei. Após administração oral dos tratamentos para camundongos por 21 dias foram realizadas coletas de sangue e determinadas as atividades de aspartato aminotransferase (AST), alanina aminotransferase (ALT), creatina quinase (CK) e as concentrações de ureia, proteína total, albumina, globulinas e frações alfa, beta e gama-globulina. A presença do tumor de Ehrlich foi responsável por aumento significativo nas atividades séricas de AST e CK e das concentrações de globulinas totais e da fração gama-globulina, além de redução dos níveis de albumina e das alfa2-globulinas. Os extratos de polissacarídeos de A. blazei reduziram as atividades séricas de AST e ALT, provavelmente devido a um efeito hepatoprotetor. Além disto, extratos de polissacarídeos e sobrenadante inibiram o aumento das gama-globulinas induzido pelo tumor. Assim, as frações de polissacarídeos e sobrenadante do extrato de A. blazei apresentam bom potencial para uso complementar ao tratamento antineoplásico.(AU)