The Influence of porcine parity on colostrum cytokine levels and their passive transfer to piglets / A influência da paridade suína na diferença dos níveis de citocinas no colostro e na transferência passiva para leitões

The limited ability of newborn piglets to produce cytokines may influence lymphocyte development and response to antigen exposure. As a result, colostrum intake is crucial because it contains nutrients that contribute to immune system development in piglets. Our goal was to investigate the effect of sow parity on the transfer of maternal cytokines to nursing piglets. Sixty piglets from nine sows were divided into six groups: piglets from gilts or sows kept with their dams and allowed to suckle normally; piglets from gilts or sows having their dams exchanged and then allowed to suckle normally; piglets from gilts or sows isolated from their dams and bottle-fed a commercial milk replacer formula for pigs. All piglets remained in the diet groups for 24 hours after birth. Concentrations of cytokines in colostrum and serum of gilt/ sows and serum of piglets were then evaluated. The 13 evaluated cytokines had higher concentrations in colostrum and serum of sows than in gilts. Concentrations of GM-CSF, IFNγ, IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-12, IL-18, and TNFα were higher in piglets suckling sows. Piglets that received commercial formula showed higher concentrations of the cytokines IL1-RA and IL-8 than piglets fed colostrum. This outcome can influence piglets' development into adulthood. In short, our findings demonstrated that maternal parity influenced colostrum cytokine composition and its maternal transfer patterns.(AU)

A capacidade limitada dos leitões recém-nascidos de produzir citocinas pode influenciar o desenvolvimento de linfócitos e a resposta à exposição ao antígeno. Portanto, a ingestão de colostro é importante porque contém nutrientes, que contribuem para o desenvolvimento do sistema imunológico do leitão. O objetivo do estudo foi investigar o efeito da paridade da porca na transferência de citocina materna para leitões lactentes. Sessenta leitões de nove porcas foram divididos em seis grupos: leitões de marrãs/porcas mantidas com suas próprias mães e amamentadas normalmente; leitões de marrãs/porcas que foram trocados de mães e amamentados normalmente; leitões de marrãs/porcas que foram isolados das mães e alimentados com mamadeira com substituto do leite para suínos. Os leitões permaneceram nos grupos por 24 horas após o nascimento. Foram avaliadas as concentrações de citocinas no colostro e plasma das marrãs/porcas e no plasma dos leitões. O colostro e o plasma das porcas apresentaram maiores concentrações das 13 citocinas analisadas do que as marrãs. No mesmo sentido, as concentrações de GM-CSF, IFNγ, IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-12, IL-18 e TNFα foram significantemente maiores nos leitões que mamaram o colostro de porcas. Os leitões que receberam fórmula comercial apresentaram, em especial, concentrações das citocinas IL1-RA e IL-8 superiores aos leitões amamentados com colostro. Isso pode influenciar o desenvolvimento até a fase adulta. Portanto, nossos dados demonstraram que a paridade materna influenciou a composição das citocinas do colostro, bem como as características das citocinas na transferência materna.(AU)

The in vitro effects of n-3 fatty acids on immune response regulation of bovine ex vivo endometrial explants / Os efeitos in vitro dos ácidos graxos n-3 na regulação da resposta imune de explantes endometriais bovinos ex vivo

Fatty acids are considered metabolic intermediaries, although new facts indicate they also work as signaling molecules with different roles in the immune response. Based on that, in this study, we investigated the anti-inflammatory effects of n-3 polyunsaturated fatty acids (PUFAs) as eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and α-linolenic acid (LNA) in ex vivo bovine endometrial explants. For this, two groups were formed: (1) LPS-challenged and (2) control, both to evaluate the accumulation of proinflammatory cytokines as interleukin 1ß (IL1B) and interleukin 6 (IL6). To develop the study, bovine female reproductive tracts from non-pregnant Angus heifers without evidence of reproductive diseases were selected. Endometrial explants were processed and treated for 24 h with EPA, DHA, and LNA in five different concentrations (0µM, 50µM, 100 µM, 200µM and 400 µM) and then, challenged with LPS for 24 h. Supernatants were collected to evaluate the concentration of IL1B and IL6 by ELISA. Explants treated with EPA from control groups reduced the concentrations of ILB (200µM) and IL6 (400 µM), and IL6 (50 µM; 100 µM) from the LPS-challenged group. DHA decreased the accumulation of IL1B and IL6 at 200 µM on explants from the LPS-challenged group, and 200 µM reduced IL6 from the control group. In contrast, explants treated with LNA only reduced the accumulation of IL1B to 400µM (from both groups). In conclusion, the EPA acid is the best anti-inflammatory option to decrease the concentration of both pro-inflammatory cytokines (IL1B and IL6) from LPS-challenged and control groups in bovine endometrial explants; while LNA evidence to be the last option to promote an anti-inflammatory response.(AU)

Os ácidos graxos são considerados intermediários metabólicos, embora novos fatos indiquem que eles também atuem como moléculas sinalizadoras com diferentes papéis na resposta imune. Dessa forma, este estudo investigou os efeitos anti-inflamatórios de ácidos graxos poliinsaturados n-3 (PUFAs) como ácido eicosapentaenóico (EPA), ácido docosahexaenóico (DHA) e ácido α-linolênico (LNA) em explantes endometriais ex vivo de bovinos. Para tal, o experimento foi divido em dois grupos: (1) Desafiado-LPS e (2) Controle, para que então pudesse avaliar o acúmulo de citocinas pró-inflamatórias como interleucina 1ß (IL1B) e interleucina 6 (IL6). Foram selecionados tratos reprodutivos de fêmeas bovinas de novilhas Angus não prenhes sem evidência de doenças reprodutivas. Explantes endometriais foram processados e tratados por 24h com EPA, DHA e LNA em cinco concentrações diferentes (0µM, 50µM, 100 µM, 200µM e 400 µM) e, em seguida, desafiados com LPS por mais 24h. Os sobrenadantes foram colhidos para avaliar a concentração de IL1B e IL6 pelo teste de ELISA. Os explantes tratados com EPA dos grupos de controle reduziram as concentrações de ILB (200 µM) e IL6 (400 µM) e no grupo desafiado com LPS houve redução das concentrações de IL6 (50 µM; 100 µM). Nos explantes do grupo desafiado com LPS, o DHA diminuiu o acúmulo de IL1B e IL6 nas concentrações de 200 µM, e no grupo controle reduziu IL6 nas concentrações de 200 µM, enquanto os explantes tratados com LNA reduziram apenas o acúmulo de IL1B a 400 µM (de ambos os grupos). Em conclusão, o ácido EPA provou ser a melhor opção anti-inflamatória para diminuir a concentração de ambas as citocinas pró-inflamatórias (IL1B e IL6) de grupos desafiados com LPS e controle em explantes endometriais bovinos; enquanto o LNA evidencia ser a opção menos viável para promover uma resposta anti-inflamatória.(AU)

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.

Innate immunity gene expression profiles in conjunctival membrane biopsies from Amazonian buffalo / Perfis de expressão de genes da imunidade inata em biopsias de membrana conjuntival de búfalos da Amazônia

Domestic buffalo production plays an economically important role in the Brazilian Amazon, but they are susceptible to many diseases favored by the tropical climate and annually flooded habitats, including ocular diseases. In this context, it is important to select genotypes that maximize innate ocular immunity in Amazonian herds. We aimed to characterise, for the first time, gene expression profiles of the innate immune system in the conjunctival membrane of buffalo. Ocular conjunctival tissue samples were collected from 60 clinically healthy slaughtered animals in the northern Brazilian state of Amapá. The samples were histologically processed for classification into three groups according to the quantitative degree of lymphoid tissue associated with the conjunctiva (discrete, G1; slight, G2; and moderate, G3 presence of lymphoid tissue). RT-PCR was used to quantify gene expression of inflammatory cytokine (IL6, IL10, TNFA, IFNG), Toll-like receptor 4 (TLR4), and Defensin beta 110 (DEFB110), relative to the endogenous GAPDH gene. G1 animals presented low expression for IL6, IL10, TNFA, and DEFB110, while G2 exhibited high expression for IL6, IL10, IFNG, and TLR4. All G3 animals showed high expression for all tested genes. These results suggest a greater resistance to pathogenic microorganisms of buffalos in the G3 group, and the proportion of lymphoid tissue associated with the conjunctiva may be related to the immune resistance of individuals.(AU)

A produção de búfalos domésticos desempenha um papel economicamente importante na Amazônia brasileira, mas eles são suscetíveis a muitas doenças favorecidas pelo clima tropical e habitats inundados anualmente, incluindo doenças oculares. Nesse contexto, é importante selecionar genótipos que maximizem a imunidade ocular inata em rebanhos amazônicos. Objetivamos caracterizar, pela primeira vez, perfis de expressão gênica do sistema imune inato na membrana conjuntival de búfalos. Amostras de tecido conjuntival ocular foram coletadas de 60 animais clinicamente saudáveis abatidos no estado do Amapá, norte do Brasil. As amostras foram processadas histologicamente para classificação em três grupos de acordo com o grau quantitativo de tecido linfoide associado à conjuntiva (discreta, G1; leve, G2; e moderada, G3 presença de tecido linfoide). RT-PCR foi utilizado para quantificar a expressão gênica de citocinas inflamatórias (IL6, IL10, TNFA, IFNG), receptor Toll-like 4 (TLR4) e Defensina beta 110 (DEFB110), em relação ao gene GAPDH endógeno. Os animais do G1 apresentaram baixa expressão para IL6, IL10, TNFA e DEFB110, enquanto G2 exibiu alta expressão para IL6, IL10, IFNG e TLR4. Todos os animais do G3 apresentaram alta expressão para todos os genes testados. Esses resultados sugerem maior resistência aos microrganismos patogênicos dos búfalos do grupo G3, e a proporção de tecido linfoide associado à conjuntiva pode estar relacionada à resistência imunológica dos indivíduos.(AU)

Characterization of macrophage polarization in lesions of dogs inoculated with Leishmania (Leishmania) infantum (BH401) strain

The aim of this work was to describe the anatomical pathology of dogs experimentally infected with Leishmania (Leishmania) infantum strain MCAN / BR / 2002 / BH401, a Brazilian form of L. infantum isolated from a symptomatic dog from an endemic area. For this, five beagles (three months old and both sexes) composed the experimental group. Markers of macrophage subpopulations M1 and M2 (related to resistance and susceptibility to visceral leishmaniasis) and the tissue cytokine transforming growth factor beta 1 (TGF-ß1) (one of the main cytokines related to the fibrosis process and anti-inflammatory action) were evaluated in livers, lungs and kidneys. The BH 401 L. infantum strain induced classical lesions of the visceral disease where all evaluated organs showed a chronic inflammatory reaction and tissue parasitism associated with a higher expression of CD163 and TGF-ß1 markers, might be related to the progression of the disease. In this work it was possible to conclude that the BH 401 strain reproduces canine visceral leishmaniasis that occurs naturally.(AU)

Detection of respiratory viruses and expression of inflammatory cytokines in patients with acute exacerbation chronic obstructive pulmonary disease in Mongolia China / Detecção de vírus respiratórios e expressão de citocinas inflamatórias em pacientes com doença pulmonar obstrutiva crônica de exacerbação aguda na Mongólia, China

Chronic obstructive pulmonary disease (COPD) was estimated to be the third cause of global mortality by 2020. Acute exacerbation COPD (AECOPD) is a sudden worsening of COPD symptoms and could be due to virus/bacterial infections and air pollution. Increased expression of inflammatory markers in patients with AECOPD is associated with viral infection. This study aimed to detect different viruses and analyze the expression of various inflammatory markers associated with AECOPD patients. Three hundred and forty-seven patients diagnosed with COPD according to GOLD criteria were included in this study. Swab samples and blood were collected for the detection of viruses by RT-PCR and expression of inflammatory markers, respectively. Of the swab samples, 113 (32.6%) of samples were positive for virus detection. Of these, HRV (39.8%) was the predominant virus detected followed by FluB (27.4%) and FluA (22.1%). The presence of HRV was significantly higher (p=0.044) among the other detected viruses. When compared to healthy controls the expression levels of TNF-α, IL-6 and IL-8 were significantly higher (p<0.05) in virus-positive patients. The IL-6 and IL-8 were the next predominantly expressed in markers among the samples. The higher expression rate of IL-8 was significantly (p<0.05) associated with patients having COPD GOLD III severity level and smoking history. Although HRV was the predominant virus detected the combined prevalence of Influenza A and B surpassing the rate of HRV. The high-level expression of well known inflammatory markers of AECOPD, TNF-α, IL-6 and IL-8 indicates a chronic severe illness. These markers play an important role and could be used as a marker for determining the severity of AECOPD.

Estima-se que a doença pulmonar obstrutiva crônica (DPOC) seja a terceira causa de mortalidade global em 2020. A exacerbação aguda DPOC (AECOPD) é um agravamento súbito dos sintomas da DPOC e pode ser devido a infecções por vírus/bactérias e poluição do ar. O aumento da expressão de marcadores inflamatórios em pacientes com AECOPD está associado à infecção viral. Este estudo teve como objetivo detectar diferentes vírus e analisar a expressão de vários marcadores inflamatórios associados a pacientes com AECOPD. Trezentos e quarenta e sete pacientes com diagnóstico de DPOC de acordo com os critérios GOLD foram incluídos neste estudo. Amostras de swab e sangue foram coletadas para detecção de vírus por RT-PCR e expressão de marcadores inflamatórios, respectivamente. Das amostras de esfregaço, 113 (32,6%) amostras foram positivas para detecção de vírus. Nestas, o HRV (39,8%) foi o vírus predominante detectado, seguido do FluB (27,4%) e do FluA (22,1%). A presença de VFC foi significativamente maior (p = 0,044) entre os demais vírus detectados. Quando comparados a controles saudáveis, os níveis de expressão de TNF-α, IL-6 e IL-8 foram significativamente maiores (p <0,05) em pacientes com vírus positivo. A IL-6 e a IL-8 foram as próximas predominantemente expressas em marcadores entre as amostras. A maior taxa de expressão de IL-8 foi significativamente (p <0,05) associada a pacientes com grau de gravidade GOLD III da DPOC e história de tabagismo. Embora o HRV tenha sido o vírus predominante, a prevalência combinada de Influenza A e B ultrapassou a taxa de HRV. O alto nível de expressão de marcadores inflamatórios bem conhecidos de AECOPD, TNF-α, IL-6 e IL-8 indica uma doença crônica grave. Esses marcadores desempenham um papel importante e podem ser usados como um marcador para determinar a gravidade da AECOPD.

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.

Anti-inflammatory activities of arthropod peptides: a systematic review

Peptides obtained from different animal species have gained importance recently due to research that aims to develop biopharmaceuticals with therapeutic potential. In this sense, arthropod venoms have drawn attention, not only because of their toxicity but mainly for the search for molecules with various bioactivities, including anti-inflammatory activity. The purpose of the present study is to gather data available in the literature on new peptides derived from arthropod species with anti-inflammatory potential. This systematic review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. Studies on peptides from arthropods that display anti-inflammatory activity were retrieved from PubMed, Scopus, Web of Science, and Google Scholar databases. The bibliographic research started in 2020 and searched papers without a limit on the publication date. The articles were analyzed using a search string containing the following terms: "Peptides" and "Anti-inflammatory", in combinations such as "Ant", "Bee", "Wasp", "Crab", "Shrimp", "Scorpion", "Spider", "Tick" and "Centipedes". Besides, a search was carried out in the databases with the terms: "Peptides", "Antitumor", or "Anticancer", and "Arthropods". Articles that met the inclusion and exclusion criteria totalized 171, and these served for data extraction. Additionally, the present review included anti-inflammatory peptides with anticancer properties. Peptides with confirmed anti-inflammatory activity were from insects (ants, bees, and wasps), crustaceans (shrimp and crabs), arachnids (scorpions, spiders, and ticks), and centipedes. These arthropod peptides act mainly by decreasing pro-inflammatory cytokines as analyzed in vitro and in vivo. Some showed significant antineoplastic activity, working in essential cellular pathways against malignant neoplasms.(AU)

Anti-inflammatory activities of arthropod peptides: a systematic review

Peptides obtained from different animal species have gained importance recently due to research that aims to develop biopharmaceuticals with therapeutic potential. In this sense, arthropod venoms have drawn attention, not only because of their toxicity but mainly for the search for molecules with various bioactivities, including anti-inflammatory activity. The purpose of the present study is to gather data available in the literature on new peptides derived from arthropod species with anti-inflammatory potential. This systematic review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. Studies on peptides from arthropods that display anti-inflammatory activity were retrieved from PubMed, Scopus, Web of Science, and Google Scholar databases. The bibliographic research started in 2020 and searched papers without a limit on the publication date. The articles were analyzed using a search string containing the following terms: "Peptides" and "Anti-inflammatory", in combinations such as "Ant", "Bee", "Wasp", "Crab", "Shrimp", "Scorpion", "Spider", "Tick" and "Centipedes". Besides, a search was carried out in the databases with the terms: "Peptides", "Antitumor", or "Anticancer", and "Arthropods". Articles that met the inclusion and exclusion criteria totalized 171, and these served for data extraction. Additionally, the present review included anti-inflammatory peptides with anticancer properties. Peptides with confirmed anti-inflammatory activity were from insects (ants, bees, and wasps), crustaceans (shrimp and crabs), arachnids (scorpions, spiders, and ticks), and centipedes. These arthropod peptides act mainly by decreasing pro-inflammatory cytokines as analyzed in vitro and in vivo. Some showed significant antineoplastic activity, working in essential cellular pathways against malignant neoplasms.(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)

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)

Markers of inflammation and insulin resistance in dogs before and after weight loss versus lean healthy dogs / Marcadores de inflamação e resistência à insulina em cães antes e depois da perda de peso versus cães saudáveis e magros

Chronic low-grade inflammation in obesity is characterized by an increased production of pro-inflammatory cytokines that contribute to insulin resistance. For this study body composition, markers of inflammation and of insulin resistance in dogs before and after weight loss were compared to those of lean dogs. Eleven client-owned obese adult dogs underwent a weight loss program with commercial dry food for weight loss and reached an ideal body condition score (BCS) six months after the beginning of the weight loss program. A Control Group of nine dogs with ideal BCS were selected for the comparison. Shapiro-Wilk test was used to test for normality, Mann Whitney were used for non-normally distributes data, and Student t-test was used for normally distributed parameters. In the Obese Group body fat decreased from 41.6% (30.7-58.6) to 29.1% (18.6-46.3) (P<0.01) and dogs maintained lean body mass throughout the weight loss program (P>0.05). Obese dogs presented higher concentration of fructosamine, triglycerides, insulin, IGF-1 and leptin than the Control Group before weight reduction (P<0.05). Serum concentrations of triglycerides, IL-2, IL-6, TNF-α, insulin, leptin and IGF-1 decreased after weight loss (P<0.01), and these concentrations were similar to the Control Group (P>0.05), except for leptin (P<0.001). No alteration on peptide YY was found. Leptin (r=0.60, P=0.01), fructosamine (r=0.44, P=0.02) and triglycerides (r=0.40, P=0.04) concentrations correlated with the reduction of body fat. Weight loss reduced the concentrations of inflammatory and insulin resistance markers and most parameters became similar to dogs that have always been lean, reinforcing the importance of weight loss in small animal practice.(AU)

A inflamação crônica de baixo grau na obesidade é caracterizada pela produção aumentada de citocinas pró-inflamatórias que contribuem para a resistência à insulina. Para este estudo a composição corporal e os marcadores de inflamação e de resistência à insulina em cães antes e após a perda de peso foram comparados aos de cães magros. Onze cães adultos obesos pertencentes a tutores foram incluídos em um programa de perda de peso com ração comercial hipocalórica e alcançaram escore de condição corporal (ECC) ideal seis meses após o início do regime. Um Grupo Controle de nove cães com ECC ideal foi selecionado para a comparação. O teste de Shapiro-Wilk foi usado para testar a normalidade, Mann Whitney foi usado para análise dos dados que não atenderam distribuição normal e o teste t de Student foi usado para parâmetros que atenderam a normalidade. No Grupo Obesidade, a gordura corporal diminuiu de 41,6% (30,7-58,6) para 29,1% (18,6-46,3) (P<0,01) e os cães mantiveram massa magra ao longo do programa de perda de peso (P>0,05). Cães obesos apresentaram maior concentração de frutosamina, triglicérides, insulina, IGF-1 e leptina do que o Grupo Controle antes da redução de peso (P<0,05). As concentrações séricas de triglicerídeos, IL-2, IL-6, TNF-α, insulina, leptina e IGF-1 diminuíram após a perda de peso (P<0,01) e, essas concentrações foram semelhantes ao Grupo Controle (P>0,05), com exceção da leptina (P<0,001). Nenhuma alteração no peptídeo YY foi encontrada. As concentrações de leptina (r=0,60; P=0,01), frutosamina (r=0,44; P=0,02) e triglicerídeos (r=0,40; P=0,04) correlacionaram-se com a redução da gordura corporal. A perda de peso reduziu as concentrações de marcadores inflamatórios e de resistência à insulina e a maioria dos parâmetros tornaram-se semelhante aos dos cães que sempre foram magros, reforçando a importância da perda de peso na prática clínica de pequenos animais.(AU)

Markers of inflammation and insulin resistance in dogs before and after weight loss versus lean healthy dogs / Marcadores de inflamação e resistência à insulina em cães antes e depois da perda de peso versus cães saudáveis e magros

Chronic low-grade inflammation in obesity is characterized by an increased production of pro-inflammatory cytokines that contribute to insulin resistance. For this study body composition, markers of inflammation and of insulin resistance in dogs before and after weight loss were compared to those of lean dogs. Eleven client-owned obese adult dogs underwent a weight loss program with commercial dry food for weight loss and reached an ideal body condition score (BCS) six months after the beginning of the weight loss program. A Control Group of nine dogs with ideal BCS were selected for the comparison. Shapiro-Wilk test was used to test for normality, Mann Whitney were used for non-normally distributes data, and Student t-test was used for normally distributed parameters. In the Obese Group body fat decreased from 41.6% (30.7-58.6) to 29.1% (18.6-46.3) (P<0.01) and dogs maintained lean body mass throughout the weight loss program (P>0.05). Obese dogs presented higher concentration of fructosamine, triglycerides, insulin, IGF-1 and leptin than the Control Group before weight reduction (P<0.05). Serum concentrations of triglycerides, IL-2, IL-6, TNF-α, insulin, leptin and IGF-1 decreased after weight loss (P<0.01), and these concentrations were similar to the Control Group (P>0.05), except for leptin (P<0.001). No alteration on peptide YY was found. Leptin (r=0.60, P=0.01), fructosamine (r=0.44, P=0.02) and triglycerides (r=0.40, P=0.04) concentrations correlated with the reduction of body fat. Weight loss reduced the concentrations of inflammatory and insulin resistance markers and most parameters became similar to dogs that have always been lean, reinforcing the importance of weight loss in small animal practice.(AU)

A inflamação crônica de baixo grau na obesidade é caracterizada pela produção aumentada de citocinas pró-inflamatórias que contribuem para a resistência à insulina. Para este estudo a composição corporal e os marcadores de inflamação e de resistência à insulina em cães antes e após a perda de peso foram comparados aos de cães magros. Onze cães adultos obesos pertencentes a tutores foram incluídos em um programa de perda de peso com ração comercial hipocalórica e alcançaram escore de condição corporal (ECC) ideal seis meses após o início do regime. Um Grupo Controle de nove cães com ECC ideal foi selecionado para a comparação. O teste de Shapiro-Wilk foi usado para testar a normalidade, Mann Whitney foi usado para análise dos dados que não atenderam distribuição normal e o teste t de Student foi usado para parâmetros que atenderam a normalidade. No Grupo Obesidade, a gordura corporal diminuiu de 41,6% (30,7-58,6) para 29,1% (18,6-46,3) (P<0,01) e os cães mantiveram massa magra ao longo do programa de perda de peso (P>0,05). Cães obesos apresentaram maior concentração de frutosamina, triglicérides, insulina, IGF-1 e leptina do que o Grupo Controle antes da redução de peso (P<0,05). As concentrações séricas de triglicerídeos, IL-2, IL-6, TNF-α, insulina, leptina e IGF-1 diminuíram após a perda de peso (P<0,01) e, essas concentrações foram semelhantes ao Grupo Controle (P>0,05), com exceção da leptina (P<0,001). Nenhuma alteração no peptídeo YY foi encontrada. As concentrações de leptina (r=0,60; P=0,01), frutosamina (r=0,44; P=0,02) e triglicerídeos (r=0,40; P=0,04) correlacionaram-se com a redução da gordura corporal. A perda de peso reduziu as concentrações de marcadores inflamatórios e de resistência à insulina e a maioria dos parâmetros tornaram-se semelhante aos dos cães que sempre foram magros, reforçando a importância da perda de peso na prática clínica de pequenos animais.(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)

TNF-alpha inhibitor adalimumab attenuates endotoxin induced cardiac damage in rats

Purpose To investigate the effects of adalimumab pretreatment on the lipopolysaccharide-mediated myocardial injury. Methods Twenty-eight Wistar rats were randomized into four groups (n=7). Control (C) group animals were injected once a day with intraperitoneal (i.p) 0.9 % saline for two days. In the Adalimumab (Ada) group, adalimumab was injected at a dose of 10 mg/kg/ day (i.p) for two days. Lipopolysaccharide (Lps) group rats were injected with a dose of 5 mg/kg (i.p) lipopolysaccharide. Lipopolysaccharide + Adalimumab (Lps+Ada) group rats received adalimumab before the administration of lipopolysaccharide. The animals were sacrificed 24 h after the last injection and blood samples were obtained for determination of biochemical cardiac injury markers and circulating levels of TNF-α and interleukin-6 (IL-6). Hearts were harvested for histological examination. Results Endotoxin exposure resulted in significant increases in serum cardiac injury markers, serum cytokines and histological myocardial injury scores in the Lps group. The levels of circulating cytokines, cardiac injury markers and histological injury scores for myocardial necrosis, perivascular cell infiltration, and inflammation were significantly reduced in Lps+Ada as compared to Lps group (p<0.05). Conclusions Adalimumab pretreatment reduces endotoxin-induced myocardial damage in rats. This beneficial effect is thought to be related to the reduction of cytokine release.(AU)

Subclinical endometritis and postpartum ovarian resumption in respect to TNF-alpha, IL-8 and CRP in Egyptian buffaloes

The study was carried out to find the relation between subclinical endometritis (SCE) and postpartum (pp.) ovarian resumption as well as to evaluate serum and endometrial TNF-α, IL-8 and serum CRP in buffaloes with and without SCE. Thirty-nine pluriparous buffaloes at the 3rd (W3), 5th (W5) and 7th (W7) week pp. were involved in this experiment. The parity of the buffaloes ranged from 4 to 8 with an average 5.8±0.2. Subclinical endometritis was diagnosed by the percentage of polymorphonuclear cells (PMNs) in uterine cytology obtained from endometrial cytobrush at W5 and W7. The cut-off point of PMNs% in buffaloes for SCE was ≥ 6% at W5 or ≥ 4% at W7. According to PMNs%, buffaloes were divided into SCE group (n=27) and non-SCE group (n=12). Ovarian cyclicity was monitored by rectal palpation, ultrasonography and progesterone assay at W3, W5 and W7. Serum and endometrial TNF-α, IL-8 and serum CRP were estimated at W5 and W7. Buffaloes with SCE (55.6%) showed delayed ovarian activity as compared to non-SCE (16.7%) animals (P=0.036). Significant increase in serum cytokines and CRP levels were detected at W5 (P ˂0.05) and W7 (P <0.01) in SCE buffaloes as compared to non-SCE. Endometrial levels of cytokines were significantly (P ˂0.05) elevated in SCE buffaloes. Serum and endometrial cytokines showed significant positive correlation. Furthermore, levels of TNF-α, IL-8 and CRP exhibited significant positive correlation with PMNs%. In conclusion, SCE delayed postpartum ovarian cyclicity in buffaloes. Moreover, TNF-α, IL-8 and CRP assessments could be efficient tools in prediction of SCE in buffaloes.

HMGB1 and inflammatory cytokines in experimental acute lung injury induced in rabbits / [HMGB1 e citocinas inflamatórias em lesão pulmonar aguda experimental induzida em coelhos]

The aim of this work was to measure HMGB1, TNF-alpha, and IL-8 in bronchoalveolar lavage (BAL), serum and TLR2 and TLR4mRNA expression in lung tissue of rabbits with two grades of acute lung injury (ALI). The animals were randomly assigned to groups with severe (S) and mild/moderate (MM) ALI, induced with warm saline, and a control group. HMGB1, TNF-alpha, IL-8, TLR2mRNA and TLR4mRNA were measured after ALI induction. The results showed increased levels of IL-8, TNF-alpha, HMGB1 and TLR4mRNA in the ALI groups. HMGB1, IL-8 and TNF-alpha concentrations in BAL were higher in S compared MM. Increased TLR4mRNA was observed in S and MM versus control. The results suggest an early participation of HMGB1 in ALI together with IL-8 and TNF-alpha and association with severity. TLR4 has early expression and role in ALI pathophysiology but is not associated with severity.(AU)

O objetivo deste trabalho é determinar os níveis de HMGB1, TNF-alfa e IL-8 no lavado broncoalveolar (BAL), bem como quantificar a expressão sérica de TLR2 e TLR4 mRNA em tecido pulmonar de coelhos com dois graus de lesão pulmonar aguda (LPA). Os animais foram distribuídos aleatoriamente em grupos com LPA grave (S) e leve / moderada (MM), induzidas com solução salina morna, e um grupo controle. HMGB1, TNF-alfa, IL-8, TLR2mRNA e TLR4mRNA foram medidos após a indução de LPA e quatro horas de ventilação mecânica. Os resultados mostraram níveis aumentados de IL-8, TNF-alfa, HMGB1 e TLR4mRNA nos grupos com LPA. As concentrações de HMGB1, IL-8 e TNF-alfa no LBA foram maiores no S comparado ao MM. Aumento de TLR4mRNA foi observado em S e MM versus controle. Os resultados sugerem uma participação precoce da HMGB1 na LPA em conjunto com IL-8 e TNF-alfa e associação com a gravidade da LPA. O TLR4 foi expresso na ALI e possivelmente possui papel precoce na fisiopatologia da LPA, mas sem associação com a gravidade.(AU)

Evaluation of cell damage and modulation of cytokines TNF-, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy / Avaliação do dano celular e da modulação das citocinas TNF-alpha, IL-6 e IL-10 em macrófagos expostos à terapia fotodinâmica mediada pela PpIX

Little is known regarding whether photodynamic therapy (PDT)-induced cell death can substantially compromise macrophages (M), which are important cells in PDT-induced immune responses. Here, parameters of PDT-mediated M cytotoxicity and cytokine production in response to protoporphyrin IX (PpIX) were evaluated. Peritoneal M from BALB/c mice were stimulated in vitro with PDT, light, PpIX, or lipopolysaccharide (LPS). After that, cell viability, lipid peroxidation, Nitric Oxide (NO), DNA damage, TNF-, IL-6 and IL-10 were evaluated. Short PDT exposure reduced cell viability by 1030%. There was a two-fold increase in NO and DNA degradation, despite the non-increase in lipoperoxidation. PDT increased TNF- and IL-10, particularly in the presence of LPS, and decreased the production of IL-6 to 10-fold. PDT causes cellular stress, induces NO radicals and leads to DNA degradation, generating a cytotoxic microenvironment. Furthermore, PDT modulates pro- and anti-inflammatory cytokines in M(AU)

Pouco se sabe se a morte celular induzida pela terapia fotodinâmica (PDT) compromete os macrófagos (M), envolvidos nas respostas imunes induzidas pela PDT. Neste estudo, foram avaliados parâmetros de citotoxicidade dos M mediada pela PDT e a produção de citocinas, frente à protoporfirina IX (PpIX). M peritoneais de camundongos BALB/c foram estimulados in vitro com PDT, luz, PpIX ou lipopolissacarídeo (LPS). Após isto, a viabilidade celular (VC), a lipoperoxidação, os níveis de óxido nítrico (NO), de DNA degradado, de TNF-, IL-6 e IL-10 foram avaliados. A exposição curta à PDT reduziu a VC em 10-30%. Os níveis de NO e de DNA degradado duplicaram, sem aumento da lipoperoxidação. Houve aumento de TNF- e IL-10, sendo maior na presença de LPS. Já a produção de IL-6 reduziu em dez vezes. A PDT induz estresse celular, gera radicais NO e causa dano ao DNA, tornando o microambiente citotóxico. Ainda, modula citocinas pró e anti-inflamatórias em M(AU)

Subclinical endometritis and postpartum ovarian resumption in respect to TNF-alpha, IL-8 and CRP in Egyptian buffaloes

The study was carried out to find the relation between subclinical endometritis (SCE) and postpartum (pp.) ovarian resumption as well as to evaluate serum and endometrial TNF-α, IL-8 and serum CRP in buffaloes with and without SCE. Thirty-nine pluriparous buffaloes at the 3rd (W3), 5th (W5) and 7th (W7) week pp. were involved in this experiment. The parity of the buffaloes ranged from 4 to 8 with an average 5.8±0.2. Subclinical endometritis was diagnosed by the percentage of polymorphonuclear cells (PMNs) in uterine cytology obtained from endometrial cytobrush at W5 and W7. The cut-off point of PMNs% in buffaloes for SCE was ≥ 6% at W5 or ≥ 4% at W7. According to PMNs%, buffaloes were divided into SCE group (n=27) and non-SCE group (n=12). Ovarian cyclicity was monitored by rectal palpation, ultrasonography and progesterone assay at W3, W5 and W7. Serum and endometrial TNF-α, IL-8 and serum CRP were estimated at W5 and W7. Buffaloes with SCE (55.6%) showed delayed ovarian activity as compared to non-SCE (16.7%) animals (P=0.036). Significant increase in serum cytokines and CRP levels were detected at W5 (P ˂0.05) and W7 (P <0.01) in SCE buffaloes as compared to non-SCE. Endometrial levels of cytokines were significantly (P ˂0.05) elevated in SCE buffaloes. Serum and endometrial cytokines showed significant positive correlation. Furthermore, levels of TNF-α, IL-8 and CRP exhibited significant positive correlation with PMNs%. In conclusion, SCE delayed postpartum ovarian cyclicity in buffaloes. Moreover, TNF-α, IL-8 and CRP assessments could be efficient tools in prediction of SCE in buffaloes.(AU)