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.

Anaplasma phagocytophilum in horses - evaluation of proinflammatory biomarkers

Background: Anaplasma phagocytophilum is an obligate intracellular pathogen transmitted by the ticks that cause equinegranulocytic anaplasmosis (EGA). This pathogen is infects predominantly blood cells, principally granulocytes and especially neutrophils. A. phagocytophilum causes an acute febrile disease in horses accompanying with lethargy, loss ofappetite, lameness and hemorrhages. In horses, this disease should be considered in all acute symptoms accompaniedby thrombocytopenia and leukopenia identified by hematological test performed. Tick-borne pathogens have becomeincreasingly threatening for both animals and also public health since ticks mostly carry numerous well-documented andundocumented pathogens, and the geographical range of ticks has expanded in the recent years. This research has aimed toevaluate the impact of A. phagocytophilum infection on some oxidative/nitrosative stress parameters, antioxidant enzymeactivities, proinflammatory biomarkers and trace element levels in horses.Materials, Methods & Results: The present study has been carried out using blood samples collected from 93 horses aged1-year and older. The blood samples were centrifuged and sera were separated. Serum samples stored in the freezer (-20°C)until the day of analysis. The DNA was extracted from blood and analysed by nested-PCR technique targeting 16S rRNAgene of A. phagocytophilum and then positive PCR products were sequenced. A. phagocytophilum was 6 horses (6.4%)showed positive nested-PCR results. An infected group comprised of 6 positive horses according to PCR analysis results also6 healthy horses as control were selected. Serum SOD (Horse Superoxide Dismutase(Cu-Zn)) ELISA Kit, MPO (ELISAAssay Kit Horse Myeloperoxidase) and GPx (Horse glutathione peroxidase 1 ELISA Kit Assay), IL1 (Horse Interleukin 1Beta ELISA Kit), IL6 (Horse Interleukin 6 ELISA Kit), TNF α (Horse Tumor Necrosis...

Wound-healing and cytokine-modulating potential of medicinal oil formulation comprising leaf extract of Murraya koenigii and olive oil / Potencial de cicatrização de feridas e modulação de citocinas de formulação de óleo medicinal compreendendo extrato de folhas de Murraya koenigii e azeite de oliva

The study investigated the wound healing effect of medicinal oil (MO) formulation prepared from Murraya koenigii leaves extract (methanolic) incorporated in olive oil. The MO was visually transparent, homogenous, smooth in texture, the viscosity grade was observed as 140 cP and easily spreadable. Pro-inflammatory cytokines IL-1ß, IL-6, and TNF-α were significantly reduced to 82.3 ± 3.5, 156 ± 6.2, 137.3. ± 5.5 pg/ml, respectively after treatment with MO when compared to disease control animals that showed IL-1ß, IL-6, and TNF-α levels of 170 ± 6, 265 ± 7, and 288.6 ± 11, pg/ml respectively. The level of pro-inflammatory cytokine in povidone iodine solution (PIS) group was 95.3 ± 3, 162 ± 6, 177.6 ± 8.9 pg/ml of IL-1ß, IL-6, and TNF-α respectively. Interestingly, the wound-healing efficacy of MO was found better as compared to povidone iodine treated standard group and concluded that MO has excellent wound healing effect.

O estudo investigou o efeito cicatrizante da formulação de óleo medicinal (MO) preparado a partir do extrato de folhas de Murraya koenigii (metanol) incorporado ao azeite de oliva. O MO era visualmente transparente, homogêneo, de textura lisa, o grau de viscosidade observado foi de 140 cP e facilmente espalhável. As citocinas pró-inflamatórias IL-1ß, IL-6 e TNF-α foram significativamente reduzidas para 82,3 ± 3,5, 156 ± 6,2, 137,3. ± 5,5 pg/ml, respectivamente, após o tratamento com MO quando comparados aos animais controle da doença que apresentaram níveis de IL-1ß, IL-6 e TNF-α de 170 ± 6, 265 ± 7 e 288,6 ± 11, pg/ml, respectivamente . O nível de citocina pró-inflamatória no grupo solução de iodopovidona (PIS) foi de 95,3 ± 3, 162 ± 6, 177,6 ± 8,9 pg/ml de IL-1ß, IL-6 e TNF-α, respectivamente. Curiosamente, a eficácia de cicatrização de feridas de MO foi encontrada melhor em comparação com o grupo padrão tratado com iodopovidona e concluiu que a preparação de MO tem efeito de cicatrização de feridas.

Through DNA sensors and hidden mitochondrial effects of SARS-CoV-2

The COVID-19 pandemic brought attention to studies about viral infections and their impact on the cell machinery. SARS-CoV-2, for example, invades the host cells by ACE2 interaction and possibly hijacks the mitochondria. To better understand the disease and to propose novel treatments, crucial aspects of SARS-CoV-2 enrolment with host mitochondria must be studied. The replicative process of the virus leads to consequences in mitochondrial function, and cell metabolism. The hijacking of mitochondria, on the other hand, can drive the extrusion of mitochondrial DNA (mtDNA) to the cytosol. Extracellular mtDNA evoke robust proinflammatory responses once detected, that may act in different pathways, eliciting important immune responses. However, few receptors are validated and are able to detect and respond to mtDNA. In this review, we propose that the mtDNA and its detection might be important in the immune process generated by SARS-CoV-2 and that this mechanism might be important in the lung pathogenesis seen in clinical symptoms. Therefore, investigating the mtDNA receptors and their signaling pathways might provide important clues for therapeutic interventions.(AU)

Through DNA sensors and hidden mitochondrial effects of SARS-CoV-2

The COVID-19 pandemic brought attention to studies about viral infections and their impact on the cell machinery. SARS-CoV-2, for example, invades the host cells by ACE2 interaction and possibly hijacks the mitochondria. To better understand the disease and to propose novel treatments, crucial aspects of SARS-CoV-2 enrolment with host mitochondria must be studied. The replicative process of the virus leads to consequences in mitochondrial function, and cell metabolism. The hijacking of mitochondria, on the other hand, can drive the extrusion of mitochondrial DNA (mtDNA) to the cytosol. Extracellular mtDNA evoke robust proinflammatory responses once detected, that may act in different pathways, eliciting important immune responses. However, few receptors are validated and are able to detect and respond to mtDNA. In this review, we propose that the mtDNA and its detection might be important in the immune process generated by SARS-CoV-2 and that this mechanism might be important in the lung pathogenesis seen in clinical symptoms. Therefore, investigating the mtDNA receptors and their signaling pathways might provide important clues for therapeutic interventions.(AU)

Changes in somatic cell count, composition and cytokine levels in milk from cows with mastitis due to mixed infections

Mastitis is a mammary gland inflammation that is very common worldwide, mostly caused by bacteria, and causes enormous economic losses. Many microorganisms cause this disease. The most common causes of mastitis by these microorganisms are Staphylococcus aureus (S. aureus), Escherichia coli (E. coli) and Streptococcus agalactiae (S. agalactiae). The anti-inflammatory properties of transforming growth factor (TGF)-ß include: 1) limiting interferon (IFN)-γ production; 2) increasing the expression of the interleukine (IL)-1 receptor antagonist; 3) inhibiting macrophage production of chemokines, pro-inflammatory cytokines, nitric oxide, and reactive oxygen intermediates; and 4) increasing macrophage clearance of bacterial debris and damaged parenchymal cells. It is stated that cytokines and milk composition change in case of mastitis. In this study, it was aimed to reveal the changes in milk TGF-ß1 and Tumor necrosis factor (TNF)-α concentrations and milk composition in mixed infections caused by three pathogens causing mastitis. In this study, milk samples from 90 cows were divided into 5 groups. Tumor necrosis factor (TNF)-α and TGF-ß1 concentrations and milk composition were determined in these milk samples. The California Mastitis Test (CMT) was applied to the cows included in the study and scoring was done. According to the CMT results of the milk samples taken, CMT(-) cows were included in group 1 (n = 22). Those with the CMT(+) were sent to the microbiology laboratory for analysis within 2 h. After the bacteria was determined, combination groupings were formed. Group 2 (n = 17), in which S. aureus and E. coli grew together, group 3 (n = 21), in which S. aureus and S. agalactiae grew together, group 4 (n = 8), in which S. agalactiae and E. coli grew together in milk samples, and milk samples without any bacterial growth in CMT (+) formed group 5 (n = 22), respectively. Somatic cell count was measured with the DeLaval Cell Counter® (Cell Counter DCC) device. Mineral matter, fat, protein, lactose, electrical conductivity and specific gravity were measured in milk samples using Lactoscan Milk Analyzer (Milkotronic/EUROPE). Milk samples were then stored at -80°C to measure TGF-ß1 and TNF-α. Tumor necrosis factor-α and TGF-ß1 concentrations in milk samples were measured using ELISA kits (Sunred Biological Technology). Changes in milk TNF-α and TGF-ß1 concentration and milk composition were determined in milk samples with mastitis caused by mixed infection. The TNF-α concentration of group 4 was higher than the other groups. On the other hand, the highest concentration of TGF-ß1 was found in group 2. While the number of somatic cells in group 1 was lower than in groups 2, 3, and 4, there was no statistical difference between groups 1 and 5. The lowest milk fat ratio was found in group 1, and it was found to be statistically lower than groups 2, 3, and 4. While the rate of solid-non-fat of group 1 increased compared to groups 2 and 3, the highest protein ratio was found in groups 1 and 5. There was no difference between the 5 groups in terms of mineral matter ratios. While the specific gravity was highest in group 1, there was no statistical difference between the other 4 groups. Overall, it was concluded that there was an increase in TNF-α and TGF-ß1 concentrations and a change in milk composition in samples with bacterial growth.(AU)

Immune response in Santa Inês sheep during Fasciola hepatica infection / Resposta imune em ovinos da raça Santa Inês durante a infecção por Fasciola hepatica

In the present study, we describe the proliferative response and the IFN-γ, IL-10, IL-4 cytokine expression profiles in peripheral blood, and IgG production, after the experimental infection of Santa Inês sheep with 250 Fasciola hepatica metacercariae that were evaluated at 60 and 210 days post-infection(dpi). The cytokine expression profile at 60 dpi was characterized by the production of IFN-γ and IL-10, which is indicative of an initial mixed Th1/Th2 response. The modulation of the response occurred at 210 dpi with a predominance of IL-10 and IL-4 over IFN-γ. Mononuclear cells from peripheral blood stimulated with F. hepatica antigen exhibited proliferative capacity at 60 dpi, whereas the response was consistent with modulation from Th1 to Th2 in the chronic phase. The IgG antibody response was more marked at 60 days than at 210 dpi, confirming the mixed-response profile. The late modulation of the T lymphocyte response in association with the predominance of IL-10 and IL-4 at 210 dpi suggests the involvement of these cytokines after the establishment of the parasites in the bile ducts. The transition from a mixed to a regulatory response in the chronic phase was also accompanied by a reduced number of eggs per gram of feces.(AU)

No presente estudo, observou-se a resposta proliferativa e os perfis de expressão de citocinas IFN-γ, IL10, IL-4 no sangue periférico, e a produção de IgG, aos 60 e 210 dias após infecção (dpi) experimental de ovinos Santa Inês com 250 metacercárias de Fasciola hepatica. O perfil de expressão de citocinas aos 60 dpi foi caracterizado pela produção de IFN-γ e IL-10, o que é indicativo de uma resposta mista inicial Th1/ Th2. A modulação da resposta ocorreu aos 210 dpi, com predominância de IL-10 e IL-4 sobre IFN-γ. As células mononucleares do sangue periférico estimuladas com antígeno total de F. hepatica adulta exibiram capacidade proliferativa aos 60 dpi, enquanto a resposta foi consistente com uma modulação de Th1 para Th2 na fase crônica. A resposta do anticorpo IgG foi maior aos 60 dpi do que aos 210 dpi. A modulação tardia da resposta dos linfócitos T em associação com a predominância de IL-10 e IL-4 aos 210 dpi sugere o envolvimento dessas citocinas após o estabelecimento dos parasitos nos ductos biliares. A transição de uma resposta mista para uma resposta com pefil regulador na fase crônica também foi acompanhada por redução dos ovos por grama de fezes.(AU)

Boar seminal plasma: current insights on its potential role for assisted reproductive technologies in swine

Seminal plasma (SP) supports not only sperm function but also the ability of spermatozoa to withstand biotechnological procedures as artificial insemination, freezing or sex sorting. Moreover, evidence has been provided that SP contains identifiable molecules which can act as fertility biomarkers, and even improve the output of assisted reproductive technologies by acting as modulators of endometrial and embryonic changes of gene expression, thus affecting embryo development and fertility beyond the sperm horizon. In this overview, we discuss current knowledge of the composition of SP, mainly proteins and cytokines, and their influence on semen basic procedures, such as liquid storage or cryopreservation. The role of SP as modulator of endometrial and embryonic molecular changes that lead to successful pregnancy will also be discussed.

Boar seminal plasma: current insights on its potential role for assisted reproductive technologies in swine

Seminal plasma (SP) supports not only sperm function but also the ability of spermatozoa to withstand biotechnological procedures as artificial insemination, freezing or sex sorting. Moreover, evidence has been provided that SP contains identifiable molecules which can act as fertility biomarkers, and even improve the output of assisted reproductive technologies by acting as modulators of endometrial and embryonic changes of gene expression, thus affecting embryo development and fertility beyond the sperm horizon. In this overview, we discuss current knowledge of the composition of SP, mainly proteins and cytokines, and their influence on semen basic procedures, such as liquid storage or cryopreservation. The role of SP as modulator of endometrial and embryonic molecular changes that lead to successful pregnancy will also be discussed.(AU)

Differential expression of T helper cytokines in the liver during early pregnancy in sheep

Liver plays important roles in the innate and adaptive immunity, and contributes to the maternal immune adjustments during pregnancy in mice and rats. T helper 1 (Th1) and Th2 cytokines are related to immune response. However, expression of Th1 and Th2 cytokines in maternal livers is unclear during early pregnancy in sheep. In this study, livers were collected on day 16 of the estrous cycle and on days 13, 16 and 25 of pregnancy (n = 6 for each group) in ewes, and qRT-PCR, western blot and immunohistochemistry were used to analyze the expression of Th1 and Th2 cytokines in the livers. Our results showed that interferon-gamma (IFN-γ), interleukin (IL)-2, IL-4, IL-6 and IL-10 were downregulated, and IL-5 was upregulated in the livers during early pregnancy. Furthermore, there was no effect for early pregnancy on expression of TNF-β in the livers, and the IFN-γ protein was limited to the endothelial cells of the proper hepatic arteries and portal veins. In conclusion, early pregnancy exerted its effect on the liver to regulate the Th cytokines expression, but there was no evident shift from Th1 to Th2 cytokines, which may be necessary for the maternal hepatic immune adjustments during early pregnancy in sheep.

Differential expression of T helper cytokines in the liver during early pregnancy in sheep

Liver plays important roles in the innate and adaptive immunity, and contributes to the maternal immune adjustments during pregnancy in mice and rats. T helper 1 (Th1) and Th2 cytokines are related to immune response. However, expression of Th1 and Th2 cytokines in maternal livers is unclear during early pregnancy in sheep. In this study, livers were collected on day 16 of the estrous cycle and on days 13, 16 and 25 of pregnancy (n = 6 for each group) in ewes, and qRT-PCR, western blot and immunohistochemistry were used to analyze the expression of Th1 and Th2 cytokines in the livers. Our results showed that interferon-gamma (IFN-γ), interleukin (IL)-2, IL-4, IL-6 and IL-10 were downregulated, and IL-5 was upregulated in the livers during early pregnancy. Furthermore, there was no effect for early pregnancy on expression of TNF-β in the livers, and the IFN-γ protein was limited to the endothelial cells of the proper hepatic arteries and portal veins. In conclusion, early pregnancy exerted its effect on the liver to regulate the Th cytokines expression, but there was no evident shift from Th1 to Th2 cytokines, which may be necessary for the maternal hepatic immune adjustments during early pregnancy in sheep.(AU)

Correlation between clinical findings, mast cell count and interleukin 31 immunostaining in the skin of dogs with atopic dermatitis / Correlação entre achados clínicos, contagem de mastócitos e imunomarcação de interleucina 31 na pele de cães com dermatite atópica

In this study the correlation between the clinical score, mast cell count and interleukin 31 (IL-31) immunostaining in the skin of dogs with atopic dermatitis was determined. A total of 31 dogs of different breeds, from one to eight years of age, were chosen for the study. The 20 females and 11 males were categorized based on the CADESI-4 system, as having discrete, moderate or marked atopic dermatitis. Skin samples were collected from the axillary and interdigital regions and stained with hematoxylin and eosin for cytohistomorphological analyses and toluidine blue to evaluate the mast cell counts, and immunohistochemistry for the IL-31 immunostaining. Animals revealing higher atopic dermatitis scores had greater numbers of mast cells and IL-31 immunolabeled cells. More numbers of cells immunolabeled for IL-31 were evident in the axillary skin compared with the interdigital skin in dogs having this condition. A correlation was identified between the clinical scores and mast cell numbers in the interdigital region, as well as between the clinical scores and number of cells immunolabeled for IL-31 in the axillary area. A correlation was also reported between the mast cell numbers and IL-31 immunolabeled cells only in the axillary skin, and none in the interdigital regions. It was thus concluded that the mast cells and IL-31 are involved in the pathogenesis of the canine atopic dermatitis (CAD), as well as lymphocytes and plasma cells. It was also observed that the higher the degree of clinical severity of the disease, the more the numbers of mast cells and IL-31 in the skin of those animals suffering from CAD, which implies the influence of these immunological constituents on the genesis of pruritus and disease progression.(AU)

Este estudo avaliou a correlação entre o escore clínico, a contagem de mastócitos e a imunomarcação de interleucina 31 (IL-31) na pele de cães com dermatite atópica. Foram selecionados 31 cães de diferentes raças, com idade entre um e oito anos, sendo 20 fêmeas e 11 machos, divididos em discretamente, moderadamente e acentuadamente acometidos por dermatite atópica segundo o sistema CADESI-4. Amostras da pele das regiões axilar e interdigital foram colhidas e submetidas às colorações de hematoxilina e eosina para a avaliação cito-histomorfológica e azul de toluidina para a contagem de mastócitos, bem como a técnica de imunoistoquímica para a imunomarcação de IL-31. Os animais com maior escore de dermatite atópica apresentaram maior número de mastócitos e de células imunomarcadas para IL-31. Houve maior número de células imunomarcadas para IL-31 na pele da axila em relação à interdigital nos cães com a doença. Foi constatada correlação entre o escore clínico e a quantidade de mastócitos no interdígito, bem como entre o escore clínico e a quantidade de células imunomarcadas para IL-31 na axila. Também foi verificada correlação entre a quantidade de mastócitos e células imunomarcadas para IL-31 na pele da região axilar, mas não da interdigital. Conclui-se que mastócitos e a IL-31 estão envolvidos na patogenia da DAC, assim como linfócitos e plasmócitos. Também, quanto maior o grau de severidade clínica da doença, maior a quantidade de mastócitos e IL-31 na pele dos animais com DAC, o que remete à influência desses componentes imunológicos na gênese do prurido e progressão da doença.(AU)

Tumor-associated macrophage is correlated with survival and SOCS protein expression in canine mammary carcinoma / Correlação do infiltrado macrofágico com a sobrevida e a expressão da proteína SOCS no carcinoma mamário canino

The inflammatory infiltrate in the tumor microenvironment, particularly in mammary tumors, has aroused great interest in oncology, to play different roles in the progression or tumor regression dependent on the types and cell subsets involved. The present study aimed to evaluate (1) the occurrence and intensity of macrophage infiltration in the mammary carcinoma microenvironment, (2) the expression of SOCS1 and SOCS3 proteins in tumor associated macrophages, (3) any association between these parameters and tumor development, as well as survival rates in female dogs. Twenty-two female dogs diagnosed as carcinoma arising in a mixed tumor (CMT) by histopathology were divided into two groups following mastectomy: dogs without metastasis (CMT(-)=11) and those with metastasis (CMT(+)=11). The following parameters were analyzed: tumor size, lymph node metastasis, clinical stage, histological grade, distribution and intensity of inflammatory infiltrate, tumor macrophage quantification by immunohistochemical analysis of SOCS1 and SOCS3 expression, and immunophenotyping of peripheral blood leukocytes by flow cytometry. Dogs with the higher proportions of macrophages in the inflammatory infiltrate (≥400/tumor) also had higher survival rates in comparison with dogs with less macrophages. Immunostaining revealed higher proportions of SOCS3-positive macrophages in dogs without lymph node metastasis, while SOCS1-positive macrophages were predominant in dogs with metastasis (p<0.05). Multivariate analysis found associations between survival rate and clinical staging (p=0.025), histological grade (p=0.007), and the expression of MHC-CI in circulating monocytes (p=0.018)...(AU)

O infiltrado inflamatório no microambiente tumoral, particularmente nos tumores mamários, tem despertado grande interesse na oncologia, por desempenhar diferentes funções na progressão ou regressão tumoral, dependendo dos tipos e subtipos celulares envolvidos. O presente estudo teve como objetivo avaliar: (1) a ocorrência e a intensidade do infiltrado macrofágico no microambiente do carcinoma mamário; (2) a expressão das proteínas SOCS1 e SOCS3 nos macrófagos associados ao tumor; (3) qualquer associação relacionada ao prognóstico entre estes parâmetros e o desenvolvimento tumoral, assim como a taxa de sobrevida. Vinte e duas cadelas diagnosticadas com carcinoma em tumor misto (CTM) por exame histopatológico foram divididas em dois grupos após a mastectomia: cadelas sem metástase (CTM(-)=11) e cadelas com metástase (CTM(+)=11). Foram analisados os seguintes parâmetros: tamanho do tumor, metástase para linfonodo, estadiamento clínico, grau histológico, distribuição e intensidade do infiltrado inflamatório, quantificação dos macrófagos tumorais por análise imuno-histoquímica da expressão de SOCS1 e SOCS3, e imunofenotipagem dos leucócitos (monócitos e linfócitos) do sangue periférico por citometria de fluxo. Cadelas que apresentavam maiores proporções de macrófagos no infiltrado inflamatório (≥400/tumor) também tiveram maior taxa de sobrevida em comparação àquelas com menos macrófagos. A imunomarcação revelou maiores proporções de macrófagos SOCS3-positivos em cães sem metástase para linfonodo, enquanto que macrófagos SOCS1-positivos foram predominantes naqueles com metástase (p<0,05). A análise multivariada identificou associações entre a taxa de sobrevida e o estadiamento clínico (p=0,025), grau histológico (p=0,007) e a expressão de MHC-CI em monócitos circulantes (p=0,018)...(AU)

Tumor-associated macrophage is correlated with survival and SOCS protein expression in canine mammary carcinoma / Correlação do infiltrado macrofágico com a sobrevida e a expressão da proteína SOCS no carcinoma mamário canino

The inflammatory infiltrate in the tumor microenvironment, particularly in mammary tumors, has aroused great interest in oncology, to play different roles in the progression or tumor regression dependent on the types and cell subsets involved. The present study aimed to evaluate (1) the occurrence and intensity of macrophage infiltration in the mammary carcinoma microenvironment, (2) the expression of SOCS1 and SOCS3 proteins in tumor associated macrophages, (3) any association between these parameters and tumor development, as well as survival rates in female dogs. Twenty-two female dogs diagnosed as carcinoma arising in a mixed tumor (CMT) by histopathology were divided into two groups following mastectomy: dogs without metastasis (CMT(-)=11) and those with metastasis (CMT(+)=11). The following parameters were analyzed: tumor size, lymph node metastasis, clinical stage, histological grade, distribution and intensity of inflammatory infiltrate, tumor macrophage quantification by immunohistochemical analysis of SOCS1 and SOCS3 expression, and immunophenotyping of peripheral blood leukocytes by flow cytometry. Dogs with the higher proportions of macrophages in the inflammatory infiltrate (≥400/tumor) also had higher survival rates in comparison with dogs with less macrophages. Immunostaining revealed higher proportions of SOCS3-positive macrophages in dogs without lymph node metastasis, while SOCS1-positive macrophages were predominant in dogs with metastasis (p<0.05). Multivariate analysis found associations between survival rate and clinical staging (p=0.025), histological grade (p=0.007), and the expression of MHC-CI in circulating monocytes (p=0.018). Higher SOCS3 expression in activated macrophages within the inflammatory infiltrate were considered indicative of an antitumor immune response, improved clinicopathological parameters and longer survival, whereas SOCS1-related activation was associated with tumor progression, metastasis development and reduced survival in female dogs with mammary carcinomas.(AU)

O infiltrado inflamatório no microambiente tumoral, particularmente nos tumores mamários, tem despertado grande interesse na oncologia, por desempenhar diferentes funções na progressão ou regressão tumoral, dependendo dos tipos e subtipos celulares envolvidos. O presente estudo teve como objetivo avaliar: (1) a ocorrência e a intensidade do infiltrado macrofágico no microambiente do carcinoma mamário; (2) a expressão das proteínas SOCS1 e SOCS3 nos macrófagos associados ao tumor; (3) qualquer associação relacionada ao prognóstico entre estes parâmetros e o desenvolvimento tumoral, assim como a taxa de sobrevida. Vinte e duas cadelas diagnosticadas com carcinoma em tumor misto (CTM) por exame histopatológico foram divididas em dois grupos após a mastectomia: cadelas sem metástase (CTM(-)=11) e cadelas com metástase (CTM(+)=11). Foram analisados os seguintes parâmetros: tamanho do tumor, metástase para linfonodo, estadiamento clínico, grau histológico, distribuição e intensidade do infiltrado inflamatório, quantificação dos macrófagos tumorais por análise imuno-histoquímica da expressão de SOCS1 e SOCS3, e imunofenotipagem dos leucócitos (monócitos e linfócitos) do sangue periférico por citometria de fluxo. Cadelas que apresentavam maiores proporções de macrófagos no infiltrado inflamatório (≥400/tumor) também tiveram maior taxa de sobrevida em comparação àquelas com menos macrófagos. A imunomarcação revelou maiores proporções de macrófagos SOCS3-positivos em cães sem metástase para linfonodo, enquanto que macrófagos SOCS1-positivos foram predominantes naqueles com metástase (p<0,05). A análise multivariada identificou associações entre a taxa de sobrevida e o estadiamento clínico (p=0,025), grau histológico (p=0,007) e a expressão de MHC-CI em monócitos circulantes (p=0,018). A maior expressão de SOCS3 nos macrófagos ativados foi considerada indicativa de uma resposta imune antitumoral, melhores parâmetros clínicos e maior taxa de sobrevida, ao passo que a ativação relacionada com SOCS1 foi associada à progressão tumoral, desenvolvimento de metástase e redução na taxa de sobrevida em cadelas com carcinoma mamário.(AU)

The effects of coenzyme Q10 on inflammation markers in streptozotocin-induced diabetic rats

Background: Coenzyme Q10 is a well-known cofactor in the mitochondrial electron transport chain required for ATP production. Coenzyme Q10 is recognized as an intracellular antioxidant that protects cell membrane phospholipids, mitochondrial membrane protein, and plasma low-density lipoprotein against oxidative damage caused by free radicals. Diabetes and its complications have been related to increased levels of free radicals and systemic proinflammatory cytokines and to an abnormal lipid profile. The aim of this study was to investigate the effects of coenzyme Q10 supplementation on some cytokine levels in streptozotocin-induced diabetic rats. Materials, Methods & Results: In this study, 38 healthy, adult male rats were used. The rats were divided into 5 groups. All animals were housed in separated cages during the four weeks. The animals in group 1 was fed standard rat pellets for 4 weeks. It was administered at 0.3 mL corn oil intraperitoneally daily for four weeks in group 2 animals. The animals in group 3 was injected intraperitoneally with 10 mg/kg CoQ10 daily for 4 weeks. Group 4 was made diabetic by subcutaneous injections of streptozotocin at dose of 40 mg/kg in 0.1 M citrate buffer (pH 4.5) single daily dose for two days and group 5 was made diabetic by subcutaneous injections of streptozotocin at dose of 40 mg/kg in 0.1 M citrate buffer (pH 4.5) single daily dose [...](AU)

The effects of coenzyme Q10 on inflammation markers in streptozotocin-induced diabetic rats

Background: Coenzyme Q10 is a well-known cofactor in the mitochondrial electron transport chain required for ATP production. Coenzyme Q10 is recognized as an intracellular antioxidant that protects cell membrane phospholipids, mitochondrial membrane protein, and plasma low-density lipoprotein against oxidative damage caused by free radicals. Diabetes and its complications have been related to increased levels of free radicals and systemic proinflammatory cytokines and to an abnormal lipid profile. The aim of this study was to investigate the effects of coenzyme Q10 supplementation on some cytokine levels in streptozotocin-induced diabetic rats. Materials, Methods & Results: In this study, 38 healthy, adult male rats were used. The rats were divided into 5 groups. All animals were housed in separated cages during the four weeks. The animals in group 1 was fed standard rat pellets for 4 weeks. It was administered at 0.3 mL corn oil intraperitoneally daily for four weeks in group 2 animals. The animals in group 3 was injected intraperitoneally with 10 mg/kg CoQ10 daily for 4 weeks. Group 4 was made diabetic by subcutaneous injections of streptozotocin at dose of 40 mg/kg in 0.1 M citrate buffer (pH 4.5) single daily dose for two days and group 5 was made diabetic by subcutaneous injections of streptozotocin at dose of 40 mg/kg in 0.1 M citrate buffer (pH 4.5) single daily dose [...]

The Effects of Coenzyme Q10 on Inflammation Markers in Streptozotocin-Induced Diabetic Rats

Background: Coenzyme Q10 is a well-known cofactor in the mitochondrial electron transport chain required for ATP production. Coenzyme Q10 is recognized as an intracellular antioxidant that protects cell membrane phospholipids, mitochondrial membrane protein, and plasma low-density lipoprotein against oxidative damage caused by free radicals. Diabetes and its complications have been related to increased levels of free radicals and systemic proinflammatory cytokines and to an abnormal lipid profile. The aim of this study was to investigate the effects of coenzyme Q10 supplementation on some cytokine levels in streptozotocin-induced diabetic rats.Materials, Methods & Results: In this study, 38 healthy, adult male rats were used. The rats were divided into 5 groups. All animals were housed in separated cages during the four weeks. The animals in group 1 was fed standard rat pellets for 4 weeks. It was administered at 0.3 mL corn oil intraperitoneally daily for four weeks in group 2 animals. The animals in group 3 was injected intraperitoneally with 10 mg/kg CoQ10 daily for 4 weeks. Group 4 was made diabetic by subcutaneous injections of streptozotocin at dose of 40 mg/kg in 0.1 M citrate buffer (pH 4.5) single daily dose for two days and group 5 was made diabetic by subcutaneous injections of streptozotocin at dose of 40 mg/kg in 0.1 M citrate buffer (pH 4.5) single daily dose

Effect of anti-TNF-alpha on the development of offspring and pregnancy loss during pregnancy in rats

Background: Etanercept binds soluble tumor necrosis factor-alpha (TNF-alpha) and is classified as pregnancy risk category B. Increase in TNF-alpha level causes preterm labour or miscarriage. Lipopolysaccharides trigger preterm birth and abortion via producing of pro-inflammatory cytokines. Cytokines are divided into two groups as pro-inflammatory and anti-inflammatory. TNF-alpha is a pro-inflammatory cytokine, whereas interleukin (IL)-10 is an anti-inflammatory cytokine. IL-10 predominant in normal pregnancy while TNF-alpha characterize in abortion and recurrent abortion. The aim of this study was to determine the effect of etanercept on the development of offspring and lipopolysaccharide-induced pregnancy loss. Materials, Methods & Results: Twenty-eight female and 7 male Wistar rats (5-6 months old) were used in this study. The rats were fed a standard pelleted diet and tap water ad libitum. After female rats were caged with males for 1 day, the presence of a vaginal plug was designated as day 0 of pregnancy. Twenty-eight pregnant Wistar rats were divided into 4 equal groups, as follows: control (0.3 mL of Normal Saline Solution intravenously on day 10 of pregnancy); etanercept (0.8 mg kg-1/day intraperitoneally on days 9 and 10 of pregnancy); lipopolysaccharide (160 µg kg-1 intravenously on day 10 of pregnancy); and etanercept + lipopolysaccharide. Blood samples were obtained from the [...](AU)

Effect of anti-TNF-alpha on the development of offspring and pregnancy loss during pregnancy in rats

Background: Etanercept binds soluble tumor necrosis factor-alpha (TNF-alpha) and is classified as pregnancy risk category B. Increase in TNF-alpha level causes preterm labour or miscarriage. Lipopolysaccharides trigger preterm birth and abortion via producing of pro-inflammatory cytokines. Cytokines are divided into two groups as pro-inflammatory and anti-inflammatory. TNF-alpha is a pro-inflammatory cytokine, whereas interleukin (IL)-10 is an anti-inflammatory cytokine. IL-10 predominant in normal pregnancy while TNF-alpha characterize in abortion and recurrent abortion. The aim of this study was to determine the effect of etanercept on the development of offspring and lipopolysaccharide-induced pregnancy loss. Materials, Methods & Results: Twenty-eight female and 7 male Wistar rats (5-6 months old) were used in this study. The rats were fed a standard pelleted diet and tap water ad libitum. After female rats were caged with males for 1 day, the presence of a vaginal plug was designated as day 0 of pregnancy. Twenty-eight pregnant Wistar rats were divided into 4 equal groups, as follows: control (0.3 mL of Normal Saline Solution intravenously on day 10 of pregnancy); etanercept (0.8 mg kg-1/day intraperitoneally on days 9 and 10 of pregnancy); lipopolysaccharide (160 µg kg-1 intravenously on day 10 of pregnancy); and etanercept + lipopolysaccharide. Blood samples were obtained from the [...]

Micropropagation of Agapanthus umbellatus var. minor by using two systems of multiplication / Micropropagação de Agapanthus umbellatus var. minor com dois sistemas de multiplicação

For conventional micropropagation methods, semisolidified medium (SM) is used; the use of this medium requires intense manipulation of the cultures and skilled labor. Systems that use liquid medium show equal or better efficiency of the multiplication process, besides reducing the cost for the elimination of agar. In this study, we evaluated the mass propagation of Agapanthus umbellatus var. minor two in vitro multiplication systems (SM system and temporary immersion system [SIT]). The plant material was grown in MS medium supplemented with 6-benzylaminopurine (6-BA; 0.0, 8.9, 17.8, and 35.6 ?M). The data obtained in this study demonstrate that the two systems used were efficient for the multiplication phase of this species. However, we recommend SIT in view of its reuse in the process of multiplication and rooting. Moreover, simple construction, low cost of the culture medium, and low cost of the bioreactors and the fact that agar is not required qualify this system as an efficient alternative for large-scale micropropagation of Agapanthus umbellatus var. minor. We recommend 17.8 ?M 6-BA for the SM system and 8.9 ?M 6-BA for SIT.

Na metodologia convencional de micropropagação utiliza-se meio nutritivo geleificado, o que acarreta intensa manipulação das culturas e envolve mão-de-obra especializada. Porém, há uma tendência em se utilizar biorreatores em meio líquido com igual ou até melhor eficiência do processo de multiplicação, além de diminuir o custo pela eliminação do ágar. O presente trabalho teve como objetivo comparar a eficiência do uso do meio geleificado (MG) e de biorreator de imersão temporária (BIT), visando a propagação em massa de Agapanthus umbellatus var. minor. O material vegetal cresceu em meio MS suplementado com diferentes concentrações de 6-BAP (0,0; 8,9; 17,8 e 35,6 ?M). As avaliações de crescimento e desenvolvimento aos 60 dias mostraram que ambos os sistemas foram eficientes na fase de multiplicação. No entanto, a imersão temporária pode ser recomendada, com vista à sua reutilização no processo de multiplicação e enraizamento. Além disso, sugere-se este sistema como uma alternativa eficiente para a micropropagação em larga escala desta espécie por sua simplicidade na construção, custo mais baixo do meio de cultura, por não utilizar ágar e pelo baixo custo dos biorreatores. Dentre as concentrações utilizadas nos sistemas, recomenda-se a concentração de 17,8 ?M de 6-BAP para o MG e 8,9 ?M de 6-BAP para o BIT.