Episcleroceratite nodular granulomatosa em cães / Nodular episclerokeratitis in dogs

Background: Episcleral inflammation may be assumed to be primary immune-mediated, secondary to intra- or extraocular diseases, or systemic abnormalities. We aimed to report a confirmed and another suspect case of nodular episclerokeratites (NEK) due to its rarity in the clinical setting and the paucity of case reports in Brazilian literature. Cases: Case 1. Refers to a 7-year-old castrated male, Collie-mixed breed, presenting with epiphora and an irregular ocular surface shape in the left eye (LE). Ophthalmic evaluation of this eye revealed mucoid discharge, conjunctival hyperemia, episcleral injection, and a gelatinous mass in the temporal limbic region. Biomicroscopic evaluation of the anterior chamber, lens, and vitreous was impaired in the LE because of corneal vessels and a mild flare in the aqueous humor. Histopathology of a scleral biopsy revealed the presence of lymphocytes, histiocytes, and some plasma cells. Positive CD3-lymphocytes were observed by immunohistochemistry, confirming the diagnosis of NEK. Case 2. Refers to a 8-year-old, spayed female Border Collie with a history of exophthalmos, conjunctival hyperemia, and inability to close the eyelid of the LE. During ophthalmic examination, an irregular espicleral nodule of approximately 9 mm was also found in the temporal limbic region, along with enlargement of episcleral vessels and scleral thinning at the equatorial region. The cornea showed mild and diffuse edema, and white crystal-like deposits were distributed in a band-like fashion at the dorsal aspect. Ultrasonography revealed scleral thinning without evidence of a mass effect arising from the iris, ciliary body, or retrobulbar space. Based on these findings, NEK was suspected. In both cases, the clinical signs reduced significantly after seven days of topical treatment with corticosteroids and cyclosporine. Discussion: It is assumed that scleral disorders are primarily immune mediated. However, such conditions may develop secondary to ocular trauma (surgery and foreign bodies), Ehrlichia canis, and Onchocerca spp. Infections and situations were ruled out in both cases. In case 1, additional histological and immunohistochemical findings supported a primary and immune-mediated scleral disease. Although the definitive diagnosis was not confirmed by histology in case 2, one can assume that the episcleral inflammation may have arisen due to an immune-mediated disorder once the eye responded positively to corticosteroid therapy. Additionally, secondary glaucoma was excluded as a possible diagnosis in case 2, because the intraocular pressure of the affected eye was below the reference range for dogs, coupled with the irregular appearance of the episclera, which is not characteristic of canine glaucoma. Moreover, in case 2, because remission of the masses of neoplastic origin after corticotherapy was not expected, the tumor was discarded. NEK has a characteristic ploriferative behavior and resistance to topical immunosuppression; clinical recurrence was not observed in the LE of either patient who remained on treatment after 60 days of follow-up. Regarding prognosis, one study showed a correlation between cellular contingent and therapeutic responses. The percentage of positive CD79a cells (B-lymphocytes) was significantly higher in cases of epicleritis and NEK, in which a poor response was achieved after topical immunosuppressive treatment. As shown by the veterinary literature and the cases described here, the complete remission of NEK is more common in unilateral cases, as confirmed after a 12-month follow-up. The 2 reported cases are useful for clarifying the common findings, diagnosis, and long-term management of NEK. Scleral abnormalities, such as NEK, must be included in the list of differential diagnoses of glaucoma, neoplasia, and endophthalmitis during ophtalmic examination.

Resposta metabólica ao trauma cirúrgico / Metabolic response to surgical trauma

Os efeitos do trauma cirúrgico no organismo, os quais são amplamente estudados pela medicina humana e veterinária, resultam em uma complexa resposta neuroendócrina e imunológica, desencadeadas para restabelecer a homeostase e preservar a vida. A resposta metabólica ao trauma é caracterizada pelo aumento da secreção de hormônios hipofisários, ativação do sistema nervoso simpático e secreção de citocinas pró-inflamatórias. Essa resposta tem por objetivo manter a perfusão tecidual, controlar a proliferação de microrganismos e mobilizar substratos para garantir a integridade das funções celulares e promover reparação tecidual. Diferentes fatores podem influenciar a intensidade da resposta do organismo ao trauma, incluindo magnitude e duração dos estímulos, enfermidades ou eventos traumáticos pré-existentes, protocolos anestésicos e técnicas cirúrgicas utilizadas. Respostas metabólicas exacerbadas ocasionam complicações pós-operatórias, tempo prolongado de recuperação dos pacientes e óbito. Através da identificação dos principais eventos relacionados à resposta metabólica ao trauma cirúrgico, é possível atuar de forma a reduzir os danos associados, melhorando a qualidade e minimizando os riscos do período pós-operatório. Sendo assim, este trabalho tem como objetivo apresentar uma revisão descrevendo a fisiopatologia e as consequências da resposta metabólica ao trauma cirúrgico, em suas diferentes formas e origens, no período perioperatório.

The effects of surgical trauma on the organism, which are widely studied by human and veterinary medicine, result in a complex neuroendocrine and immune responses triggered to restore homeostasis and preserve life. The metabolic response to trauma is characterized by increased secretion of hypophyseal hormones, activation of the sympathetic nervous system, and secretion of pro-inflammatory cytokines. This response aims to maintain tissue perfusion, control the proliferation of microorganisms, and mobilize substrates to ensure the integrity of cellular functions and promote tissue repair. Different factors can influence the intensity of the organism's response to trauma, including magnitude and duration of stimuli, pre-existing infirmities or traumas, anaesthetic protocols, and surgical techniques applied. Exacerbated metabolic responses can cause post-operative complications, prolonged recovery time, and death. By identifying the main events related to the metabolic response to surgical trauma, it is possible to act to reduce the associated damages, improving quality and minimizing the risks of the postoperative period. Thus, the present paper aims to present a review describing the physiopathology and the consequences of the metabolic response to surgical trauma, in its different forms and origins, in the perioperative period.

Effects of ß-glucan from S. cerevisiae on the expression of Casp9, Ccna2 and Sod1 genes in MCF-7 cells

ß-Glucans (ßG) are polysaccharides widely distributed in nature with chemopreventive properties. The aim of this study was to investigate the effects of ßG and the combined treatment with doxorubicin (Dox) on cell viability and mRNA levels of genes involved in cell cycle, apoptosis and antioxidant response. ßG was not cytotoxic. The mRNA levels of CCNA2of cells exposed to ß-glucan was upregulated and the exposure to Dox decreased the expression, while the combination led to an upregulation. Modulation of mRNA levels of CASP9suggest that ßG could inhibit promotion and progression steps of carcinogenesis, eliminatingneoplastic cells. The upregulation of CCNA2gene in combined treatment could be occurred due to ability of ßG in restoring the cell cycle distribution pattern after treatment with Dox. The upregulation of SOD1suggests that ßG can enhance the intracellular antioxidant defense, reducing the levels of superoxide dismutase induced by Dox. This response could reduce oxidative damage and attenuate tissue damage during chemotherapeutic treatment. Our data suggest that the drug combination may be less effective in killing tumor cells than the treatment with Dox alone. Thus, future studies should carefully consider this effect on indication of ßG during chemotherapy.Keywords:caspase-9; cyclin A2; superoxide dismutase 1; cell cycle; antioxidant.Received on July 2, 2020.Accepted on February 7, 2022.IntroductionGlucans are polysaccharides widely distributed in nature and oftenstudied due to chemopreventive properties. They are constituent of the cell wall of plants (oats and barley), algae, bacteria and fungi. ß-glucans (ßG)have a common structure comprising a main chain of ß-(1,3) and/or ß-(1,4) D-glucopyranosyl unit and they differ in length and branching structures. ßG of Saccharomyces cerevisiaehave 1→6 side branches while those of bacteria have 1→4 side branches (Chan, Chan, & Sze, 2009). ßGcan prevent DNA damage induced by chemical and physical agents (Ghavami,Goliaei, Taghizadeh, & Nikoofar, 2014). Some authors showed its significant efficacy in preventing mutagenic effects caused by doxorubicin, cyclophosphamide and cisplatin (Tohamy, El-Ghor, El-Nahas, & Noshy, 2003), methyl methanesulfonate (Oliveira et al., 2007)and hydrogen peroxide (Slamenová, 2003). Moreover, some studies have related the antioxidant ability of ßGagainst reactive free radicals formed by endogenous metabolic processes or exogenous chemicals (Tsiapali et al., 2001; Slamenová,2003; Sener, Eksioglu-Demiralp, Cetiner, Ercan, & Yegen, 2006; Guerra Dore et al., 2007; Kofuji et al., 2012; Lei et al., 2015). Yeast-derived ßGhave modulating action of humoral and cellular immune responses (Vetvicka et al., 2007).This activity provides protection to the organism against infections and cancer development (Samuelsen, Schrezenmeir, & Knutsen, 2014; Roudbary, Daneshmand, Hajimorad, Roudbarmohammadip, & Hassan, 2015). Despite postulated modes of action by which ß-glucan works are lacking information about the molecular mechanisms involved in the chemopreventive activity of this polysaccharide. In addition, compounds with chemopreventive properties can contribute to reduce side effects and toxicity during the chemotherapeutic treatment. Therefore, the aim of this study was to investigate the effects of ßG and the combined treatment with doxorubicin (Dox) on the expression of genes related with apoptosis (CASP9), cell cycle control (CCNA2)and antioxidant defense (SOD1)in human breast cancer MCF-7 cells. Doxorubicin (Dox) was chosen because it is one of the most used chemotherapeutic agent for cancer treatment. The limitation on the use of Dox in cancer treatment is the lack of selectivity against cancer cells and, consequently, its toxicity to patients.(AU)

In situ cellular immune response in non-ulcerated skin lesions due to Leishmania (L) infantum chagasi infection

Skin lesions of patients affected by non-ulcerated cutaneous leishmaniasis (NUCL) caused by L. (L.) infantum chagasi are characterized by lymphohistiocytic inflammatory infiltrate associated with epithelioid granuloma and scarce parasitism. However, the in situ cellular immune response of these patients is unclear. Therefore, the aim of the present study was to characterize the cellular immune response in the skin lesions of patients affected by NUCL. Methods Twenty biopsies were processed by immunohistochemistry using primary antibodies to T lymphocytes (CD4, CD8), NK cells, B lymphocytes, macrophages, nitric oxide synthase and interferon-gamma. Results Immunohistochemistry revealed higher expression of all cellular types and molecules (IFN-γ, iNOS) in the dermis of diseased skin compared to the skin of healthy individuals (p < 0.05). Morphometric analysis performed in the skin lesions sections showed the predominance of CD8+ T lymphocytes in the mononuclear infiltrate, followed by macrophages, mostly iNOS+, a response that could be mediated by IFN-γ. Conclusion Our study improves knowledge of the cellular immune response in non-ulcerated or atypical cutaneous leishmaniasis caused by L. (L.) infantum chagasi in Central America and pointed to the pivotal participation of CD8+ T lymphocytes in the host defense mechanisms against the parasite in patients with NUCL.(AU)

In situ cellular immune response in non-ulcerated skin lesions due to Leishmania (L) infantum chagasi infection

Skin lesions of patients affected by non-ulcerated cutaneous leishmaniasis (NUCL) caused by L. (L.) infantum chagasi are characterized by lymphohistiocytic inflammatory infiltrate associated with epithelioid granuloma and scarce parasitism. However, the in situ cellular immune response of these patients is unclear. Therefore, the aim of the present study was to characterize the cellular immune response in the skin lesions of patients affected by NUCL. Methods Twenty biopsies were processed by immunohistochemistry using primary antibodies to T lymphocytes (CD4, CD8), NK cells, B lymphocytes, macrophages, nitric oxide synthase and interferon-gamma. Results Immunohistochemistry revealed higher expression of all cellular types and molecules (IFN-γ, iNOS) in the dermis of diseased skin compared to the skin of healthy individuals (p < 0.05). Morphometric analysis performed in the skin lesions sections showed the predominance of CD8+ T lymphocytes in the mononuclear infiltrate, followed by macrophages, mostly iNOS+, a response that could be mediated by IFN-γ. Conclusion Our study improves knowledge of the cellular immune response in non-ulcerated or atypical cutaneous leishmaniasis caused by L. (L.) infantum chagasi in Central America and pointed to the pivotal participation of CD8+ T lymphocytes in the host defense mechanisms against the parasite in patients with NUCL.(AU)

Inflammation: improving understanding to prevent or ameliorate kidney diseases

Abstract Inflammatory processes are believed to play an important role in immune response to maintain tissue homeostasis by activating cellular signaling pathways and releasing inflammatory mediators in the injured tissue. Although acute inflammation can be considered protective, an uncontrolled inflammation may evolve to tissue damage, leading to chronic inflammatory diseases. Inflammation can be considered the major factor involved in the pathological progression of acute and chronic kidney diseases. Functional characteristics of this organ increase its vulnerability to developing various forms of injuries, including acute kidney injury (AKI) and chronic kidney disease (CKD). In view of translational research, several discoveries should be considered regarding the pathogenesis of the inflammatory process, which results in the validation of biomarkers for early detection of kidney diseases. Biomarkers enable the identification of proinflammatory mediators in kidney affections, based on laboratory research applied to clinical practice. Some inflammatory molecules can be useful biomarkers for the detection and diagnosis of kidney diseases, such as neutrophil gelatinase-associated lipocalin, kidney injury molecule-1 and interleukin 18.

Inflammation: improving understanding to prevent or ameliorate kidney diseases

Inflammatory processes are believed to play an important role in immune response to maintain tissue homeostasis by activating cellular signaling pathways and releasing inflammatory mediators in the injured tissue. Although acute inflammation can be considered protective, an uncontrolled inflammation may evolve to tissue damage, leading to chronic inflammatory diseases. Inflammation can be considered the major factor involved in the pathological progression of acute and chronic kidney diseases. Functional characteristics of this organ increase its vulnerability to developing various forms of injuries, including acute kidney injury (AKI) and chronic kidney disease (CKD). In view of translational research, several discoveries should be considered regarding the pathogenesis of the inflammatory process, which results in the validation of biomarkers for early detection of kidney diseases. Biomarkers enable the identification of proinflammatory mediators in kidney affections, based on laboratory research applied to clinical practice. Some inflammatory molecules can be useful biomarkers for the detection and diagnosis of kidney diseases, such as neutrophil gelatinase-associated lipocalin, kidney injury molecule-1 and interleukin 18.(AU)

Inflammation: improving understanding to prevent or ameliorate kidney diseases

Inflammatory processes are believed to play an important role in immune response to maintain tissue homeostasis by activating cellular signaling pathways and releasing inflammatory mediators in the injured tissue. Although acute inflammation can be considered protective, an uncontrolled inflammation may evolve to tissue damage, leading to chronic inflammatory diseases. Inflammation can be considered the major factor involved in the pathological progression of acute and chronic kidney diseases. Functional characteristics of this organ increase its vulnerability to developing various forms of injuries, including acute kidney injury (AKI) and chronic kidney disease (CKD). In view of translational research, several discoveries should be considered regarding the pathogenesis of the inflammatory process, which results in the validation of biomarkers for early detection of kidney diseases. Biomarkers enable the identification of proinflammatory mediators in kidney affections, based on laboratory research applied to clinical practice. Some inflammatory molecules can be useful biomarkers for the detection and diagnosis of kidney diseases, such as neutrophil gelatinase-associated lipocalin, kidney injury molecule-1 and interleukin 18.(AU)

Analysis of systemic and cutaneous inflammatory immune response in canine atopic dermatitis

Background: Canine atopic dermatitis (CAD) is a chronic and inflammatory disease present in veterinary dermatologicalpractice. The inflammation in CAD is triggered by environmental allergens and skin microorganisms, which are responsiblefor the worsening of cutaneous lesions. This continuous activation of skin inflammatory process can induce the productionof free radicals that also contribute to cellular damage and ultimately leads to changes in blood parameters in dogs withCAD. Although there are reports of inflammatory parameters in CAD, there are a lack of studies correlating skin lesions,blood leukocytes and oxidative stress. Based on that, this study aimed to evaluate the integumentary and systemic inflammatory response in dogs with atopic dermatitis.Materials, Methods & Results: Dogs with confirmed diagnosis of canine atopic dermatitis (n = 10) were divided in twogroups according to CADESI-IV: AI, with CADESI between 0-10, AII, with CADESI between 10-34, and control group (n= 5). Blood-biochemical and histological analysis were performed to access systemic and cutaneous inflammatory response.AII group tended to higher neutrophil and eosinophil counts, as well as neutrophil/lymphocyte ratio (NLR) when comparedto AI. The albumin was lower in AII compared to AI and control (P < 0.05), while total bilirubin and malondialdehyde(MDA) did not differ between groups. NLR (r = 0.64 and P = 0.04) and MDA (r = 0.54 and P = 0.1) were positively correlated with CADESI, while albumin was negatively correlated with CADESI (r = -0.79 and P = 0.005). Histopathologicalanalysis revealed a larger number of neutrophils, macrophages and mast cells in AI and AII than in control group (P < 0.05).Discussion: In this study it was possible to evaluate the systemic and cutaneous leukocyte dynamics in CAD. Skin inflammation induces the production of chemotactic molecules contribute to neutrophil outflow from blood vessel toward the...

Analysis of systemic and cutaneous inflammatory immune response in canine atopic dermatitis

Background: Canine atopic dermatitis (CAD) is a chronic and inflammatory disease present in veterinary dermatologicalpractice. The inflammation in CAD is triggered by environmental allergens and skin microorganisms, which are responsiblefor the worsening of cutaneous lesions. This continuous activation of skin inflammatory process can induce the productionof free radicals that also contribute to cellular damage and ultimately leads to changes in blood parameters in dogs withCAD. Although there are reports of inflammatory parameters in CAD, there are a lack of studies correlating skin lesions,blood leukocytes and oxidative stress. Based on that, this study aimed to evaluate the integumentary and systemic inflammatory response in dogs with atopic dermatitis.Materials, Methods & Results: Dogs with confirmed diagnosis of canine atopic dermatitis (n = 10) were divided in twogroups according to CADESI-IV: AI, with CADESI between 0-10, AII, with CADESI between 10-34, and control group (n= 5). Blood-biochemical and histological analysis were performed to access systemic and cutaneous inflammatory response.AII group tended to higher neutrophil and eosinophil counts, as well as neutrophil/lymphocyte ratio (NLR) when comparedto AI. The albumin was lower in AII compared to AI and control (P < 0.05), while total bilirubin and malondialdehyde(MDA) did not differ between groups. NLR (r = 0.64 and P = 0.04) and MDA (r = 0.54 and P = 0.1) were positively correlated with CADESI, while albumin was negatively correlated with CADESI (r = -0.79 and P = 0.005). Histopathologicalanalysis revealed a larger number of neutrophils, macrophages and mast cells in AI and AII than in control group (P < 0.05).Discussion: In this study it was possible to evaluate the systemic and cutaneous leukocyte dynamics in CAD. Skin inflammation induces the production of chemotactic molecules contribute to neutrophil outflow from blood vessel toward the...(AU)

Efeito dos anticorpos maternos sobre a resposta imune induzida pela vacinação em bezerros Holandeses / Effect of maternal antibodies on vaccine-induced immune responses in Holstein calves

Objetivou-se avaliar o efeito dos anticorpos (ACs) maternos sobre resposta imune humoral induzida pela vacinação em bezerros Holandeses. Bezerros foram distribuídos aleatoriamente em quatro grupos: G1 - vacinados no D14 e D44 (n=6); G2 - vacinados no D90 e D120 (n=5); G3 - vacinados no D180 e D210 (n=8); controle: não vacinado (n=5). Utilizaram-se 5mL de vacina comercial (Cattle Master Gold FP5+L5® - Zoetis, Brasil), por via subcutânea. Foi realizada vírus neutralização (VN) no momento da vacinação, booster e 30 dias após a revacinação. Não foram observadas diferenças entre controle e G1 ou G2 para a frequência de soropositivos ou títulos de ACs contra os vírus respiratórios (P≥0,05). G3 apresentou maior produção de ACs em relação ao controle para BoHV-1 (P<0,01), BRSV (P<0,01) e BPIV-3 (P=0,02) após o booster (D240). A análise no tempo também demonstrou aumento nos títulos de ACs no G3 (P≤0,05). O perfil clínico revelou broncopneumonia apenas no grupo controle (n=4/5) entre 80-135 dias de vida. A imunidade colostral e a vacinal apresentaram perfis inversamente proporcionais, com maior produção de ACs aos seis meses de idade. Devido à precocidade da doença respiratória, estudos complementares são necessários para esclarecer o papel da resposta imune celular na vacinação diante dos ACs maternos.(AU)

This research aimed to evaluate the effect of colostral antibodies (ABs) on the humoral immune response induced by vaccination in Holstein calves. Twenty-four calves were randomly assigned into four groups: G1 - vaccinated on D14 and D44 (n= 6); G2 - on D90 and D120 (n= 5); G3 - on D180 and D210 (n= 8); Control: unvaccinated (n= 5). Commercial vaccine (Cattle Master Gold FP5+L5® - Zoetis, Brazil) was administered subcutaneously (5mL). Virus neutralization test (VN) was performed at the time of vaccination, booster and 30 days after booster to determine AB titers. No differences were observed between control and G1 or G2 for seropositive frequencies and ABs titers (P≥ 0.05). G3 showed higher AB production than control for BoHV-1 (P< 0.01), BRSV (P< 0.01) and BPIV-3 (P= 0.02) after booster (D240). Overtime analysis also exhibited increase in AB titers in G3 (P≤ 0,05). Bronchopneumonia was identified in the control group (n= 4/5) between 80-135 days of life. The colostral and vaccinal immunity presented inversely proportional profiles, with higher production of ABs at 6 months of age. Due to the precocity of respiratory disease further studies are required to clarify the role of cellular immune response to vaccination in face of maternal ABs.(AU)

Efeito dos anticorpos maternos sobre a resposta imune induzida pela vacinação em bezerros Holandeses / Effect of maternal antibodies on vaccine-induced immune responses in Holstein calves

Objetivou-se avaliar o efeito dos anticorpos (ACs) maternos sobre resposta imune humoral induzida pela vacinação em bezerros Holandeses. Bezerros foram distribuídos aleatoriamente em quatro grupos: G1 - vacinados no D14 e D44 (n=6); G2 - vacinados no D90 e D120 (n=5); G3 - vacinados no D180 e D210 (n=8); controle: não vacinado (n=5). Utilizaram-se 5mL de vacina comercial (Cattle Master Gold FP5+L5® - Zoetis, Brasil), por via subcutânea. Foi realizada vírus neutralização (VN) no momento da vacinação, booster e 30 dias após a revacinação. Não foram observadas diferenças entre controle e G1 ou G2 para a frequência de soropositivos ou títulos de ACs contra os vírus respiratórios (P≥0,05). G3 apresentou maior produção de ACs em relação ao controle para BoHV-1 (P<0,01), BRSV (P<0,01) e BPIV-3 (P=0,02) após o booster (D240). A análise no tempo também demonstrou aumento nos títulos de ACs no G3 (P≤0,05). O perfil clínico revelou broncopneumonia apenas no grupo controle (n=4/5) entre 80-135 dias de vida. A imunidade colostral e a vacinal apresentaram perfis inversamente proporcionais, com maior produção de ACs aos seis meses de idade. Devido à precocidade da doença respiratória, estudos complementares são necessários para esclarecer o papel da resposta imune celular na vacinação diante dos ACs maternos.(AU)

This research aimed to evaluate the effect of colostral antibodies (ABs) on the humoral immune response induced by vaccination in Holstein calves. Twenty-four calves were randomly assigned into four groups: G1 - vaccinated on D14 and D44 (n= 6); G2 - on D90 and D120 (n= 5); G3 - on D180 and D210 (n= 8); Control: unvaccinated (n= 5). Commercial vaccine (Cattle Master Gold FP5+L5® - Zoetis, Brazil) was administered subcutaneously (5mL). Virus neutralization test (VN) was performed at the time of vaccination, booster and 30 days after booster to determine AB titers. No differences were observed between control and G1 or G2 for seropositive frequencies and ABs titers (P≥ 0.05). G3 showed higher AB production than control for BoHV-1 (P< 0.01), BRSV (P< 0.01) and BPIV-3 (P= 0.02) after booster (D240). Overtime analysis also exhibited increase in AB titers in G3 (P≤ 0,05). Bronchopneumonia was identified in the control group (n= 4/5) between 80-135 days of life. The colostral and vaccinal immunity presented inversely proportional profiles, with higher production of ABs at 6 months of age. Due to the precocity of respiratory disease further studies are required to clarify the role of cellular immune response to vaccination in face of maternal ABs.(AU)

Marbofloxacin induces leishmanicidal activity and less inflammatory response in Leishmania chagasi infected macrophages / Marbofloxacina induz atividade leishmanicida e menos resposta inflamatória em macrófagos infectados por Leishmania chagasi / Marbofloxacino induce actividad leishmanicida y menor respuesta inflamatoria en macrófagos infectados con Leishmania chagasi

Marbofloxacin has promising leishmanicidal activity due to the direct action on the Leishmania chagasi amastigotes. It was developed only for veterinary medicine uses, and it could be used as an drug for the treatment of Canine Visceral Leishmaniasis (CVL).In the present study, we evaluated the leishmanicidal effect and macrophage modulation of marbofloxacin in macrophages infected with amastigotes of L. chagasi. Macrophages were collected from peritoneum of BALB/c mice and infected with promastigotes of L. chagasi. After internalization and transformation into amastigotes forms, cells were treated with marbofloxacinin concentration of100, 500 and 750 μg/mL for 18 hours. The leishmanicidal effect was evaluated by morphological spects of amastigotes inside of macrophages by phagocytosis assay and detection of death amastigotes and macrophages by TUNEL assay. Macrophage modulation was evaluated by release of cytotoxic metabolites and cytokine production. The results showed that L. chagasi-infected macrophages treated with the highest concentration of the drug showed lower amount of amastigotes into the macrophages (p<0,0632) than untreated infected cells. There was pronounced presence of cellular vacuoles in the treated infected-cells, and more apoptotic amastigotes in alive macrophages. It was observed decreased levels of H2O2, IL-1β, IL-6, and TNF-αdose-dependent of marbofloxacin in infected macrophages. The results indicate the leishmanicidal effect of marbofloxacin in infected macrophages and immunomodulation actions in these cells.

A marbofloxacina tem promissora atividade leishmanicida devido à ação direta sobre as amastigotas de Leishmania chagasi. Foi desenvolvido apenas para uso em medicina veterinária, podendo ser utilizado no tratamento da Leishmaniose Visceral Canina (LVC). No presente estudo, avaliamos o efeito leishmanicida e a modulação macrofágica da marbofloxacina em macrófagos infectados com amastigotas de L. chagasi. Os macrófagos foram coletados do peritônio de camundongos BALB/c infectados com promastigotas de L. chagasi. Após internalização e transformação em formas amastigotas, as células foram tratadas com marbofloxacina nas concentrações 100, 500 e 750μg / mL por 18 horas. O efeito leishmanicida foi avaliado pelos aspectos morfológicos de amastigotas dentro de macrófagos, por ensaio de fagocitose e detecção de amastigotas e macrófagos mortos por ensaio TUNEL. A modulação dos macrófagos foi avaliada pela liberação de metabólitos citotóxicos e produção de citocinas. Os resultados mostraram que macrófagos infectados com L. chagasi tratados com a maior concentração da droga apresentaram menor quantidade de amastigotas nos macrófagos (p <0,0632) do que células infectadas não tratadas. Houve a presença pronunciada de vacúolos celulares nas células infectadas tratadas, e mais amastigotas apoptóticas em macrófagos vivos. Observou-se diminuição dos níveis de H2O2, IL-1β, IL-6 e TNF-αdependente da dose de marbofloxacina em macrófagos infectados. Os resultados indicam o efeito leishmanicida da marbofloxacina em macrófagos infectados e ações imunomoduladoras nessas células.

El marbofloxacino tiene una actividad leishmanicida prometedora debido a la acción directa sobre amastigotes de Leishmania chagasi. Fue desarrollado solo para uso en medicina veterinaria y puede usarse en el tratamiento de la leishmaniasis visceral canina (LVC). En el presente estudio, evaluamos el efecto leishmanicida y la modulación de macrófagosde marbofloxacino en macrófagos infectados con amastigotes de L. chagasi. Se recogieron macrófagos del peritoneo de ratones BALB/c infectados con promastigotes de L. chagasi. Despuésde la internalizacion y transformacion en formas amastigotes, las células se trataron con marbofloxacina a concentraciones de 100, 500 y 750 μg / ml durante 18 horas. El efecto leishmanicida se evaluó mediante los aspectos morfológicosde amastigotes dentro de macrófagos, mediante ensayo de fagocitosis y detección de amastigotes y macrófagos muertos mediante ensayo TUNEL. La modulación de los macrófagos se evaluó mediante la liberación de metabolitos citotóxicos y la producción de citocinas. Los resultados mostraron que los macrófagos infectados con L. chagasi tratados con la concentración más altas del fármaco tenían una menor cantidad de amastigotes en los macrófagos (p <0,0632) que las células infectadas no tratadas. Hubo una presencia pronunciada de vacuolas celulares en las células infectadas tratadas y más amastigotes apoptóticas en macrófagos vivos. Se observó una disminuicón en los niveles de H2O2, IL-1β, IL-6 y TNF-α dependiendo de la dosis de marbofloxacino en macrófagos infectados. Los resultados indican el efecto leishmanicida de el marbofloxacino en macrófagos infectados y acciones inmunomoduladoras en estas células.

Marbofloxacin induces leishmanicidal activity and less inflammatory response in Leishmania chagasi infected macrophages / Marbofloxacina induz atividade leishmanicida e menos resposta inflamatória em macrófagos infectados por Leishmania chagasi / Marbofloxacino induce actividad leishmanicida y menor respuesta inflamatoria en macrófagos infectados con Leishmania chagasi

Marbofloxacin has promising leishmanicidal activity due to the direct action on the Leishmania chagasi amastigotes. It was developed only for veterinary medicine uses, and it could be used as an drug for the treatment of Canine Visceral Leishmaniasis (CVL).In the present study, we evaluated the leishmanicidal effect and macrophage modulation of marbofloxacin in macrophages infected with amastigotes of L. chagasi. Macrophages were collected from peritoneum of BALB/c mice and infected with promastigotes of L. chagasi. After internalization and transformation into amastigotes forms, cells were treated with marbofloxacinin concentration of100, 500 and 750 μg/mL for 18 hours. The leishmanicidal effect was evaluated by morphological spects of amastigotes inside of macrophages by phagocytosis assay and detection of death amastigotes and macrophages by TUNEL assay. Macrophage modulation was evaluated by release of cytotoxic metabolites and cytokine production. The results showed that L. chagasi-infected macrophages treated with the highest concentration of the drug showed lower amount of amastigotes into the macrophages (p<0,0632) than untreated infected cells. There was pronounced presence of cellular vacuoles in the treated infected-cells, and more apoptotic amastigotes in alive macrophages. It was observed decreased levels of H2O2, IL-1β, IL-6, and TNF-αdose-dependent of marbofloxacin in infected macrophages. The results indicate the leishmanicidal effect of marbofloxacin in infected macrophages and immunomodulation actions in these cells.(AU)

A marbofloxacina tem promissora atividade leishmanicida devido à ação direta sobre as amastigotas de Leishmania chagasi. Foi desenvolvido apenas para uso em medicina veterinária, podendo ser utilizado no tratamento da Leishmaniose Visceral Canina (LVC). No presente estudo, avaliamos o efeito leishmanicida e a modulação macrofágica da marbofloxacina em macrófagos infectados com amastigotas de L. chagasi. Os macrófagos foram coletados do peritônio de camundongos BALB/c infectados com promastigotas de L. chagasi. Após internalização e transformação em formas amastigotas, as células foram tratadas com marbofloxacina nas concentrações 100, 500 e 750μg / mL por 18 horas. O efeito leishmanicida foi avaliado pelos aspectos morfológicos de amastigotas dentro de macrófagos, por ensaio de fagocitose e detecção de amastigotas e macrófagos mortos por ensaio TUNEL. A modulação dos macrófagos foi avaliada pela liberação de metabólitos citotóxicos e produção de citocinas. Os resultados mostraram que macrófagos infectados com L. chagasi tratados com a maior concentração da droga apresentaram menor quantidade de amastigotas nos macrófagos (p <0,0632) do que células infectadas não tratadas. Houve a presença pronunciada de vacúolos celulares nas células infectadas tratadas, e mais amastigotas apoptóticas em macrófagos vivos. Observou-se diminuição dos níveis de H2O2, IL-1β, IL-6 e TNF-αdependente da dose de marbofloxacina em macrófagos infectados. Os resultados indicam o efeito leishmanicida da marbofloxacina em macrófagos infectados e ações imunomoduladoras nessas células.(AU)

El marbofloxacino tiene una actividad leishmanicida prometedora debido a la acción directa sobre amastigotes de Leishmania chagasi. Fue desarrollado solo para uso en medicina veterinaria y puede usarse en el tratamiento de la leishmaniasis visceral canina (LVC). En el presente estudio, evaluamos el efecto leishmanicida y la modulación de macrófagosde marbofloxacino en macrófagos infectados con amastigotes de L. chagasi. Se recogieron macrófagos del peritoneo de ratones BALB/c infectados con promastigotes de L. chagasi. Despuésde la internalizacion y transformacion en formas amastigotes, las células se trataron con marbofloxacina a concentraciones de 100, 500 y 750 μg / ml durante 18 horas. El efecto leishmanicida se evaluó mediante los aspectos morfológicosde amastigotes dentro de macrófagos, mediante ensayo de fagocitosis y detección de amastigotes y macrófagos muertos mediante ensayo TUNEL. La modulación de los macrófagos se evaluó mediante la liberación de metabolitos citotóxicos y la producción de citocinas. Los resultados mostraron que los macrófagos infectados con L. chagasi tratados con la concentración más altas del fármaco tenían una menor cantidad de amastigotes en los macrófagos (p <0,0632) que las células infectadas no tratadas. Hubo una presencia pronunciada de vacuolas celulares en las células infectadas tratadas y más amastigotes apoptóticas en macrófagos vivos. Se observó una disminuicón en los niveles de H2O2, IL-1β, IL-6 y TNF-α dependiendo de la dosis de marbofloxacino en macrófagos infectados. Los resultados indican el efecto leishmanicida de el marbofloxacino en macrófagos infectados y acciones inmunomoduladoras en estas células.(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)

Phenoloxidase activity and haemolymph cytology in honeybees challenged with a virus suspension (deformed wings virus DWV) or phosphate buffered suspension (PBS) / Atividade fenoloxidase e citologia de hemolinfa em abelhas infectadas com uma suspensão de vírus (DWV deformed wings) ou suspensão tamponada com fosfato (PBS)

The innate immune system of honeybees mainly consists in antimicrobial peptides, cellular immunity and melanisation. In order to investigate the immune response of honeybees to immune stressors, three stress degrees were tested. Newly emerged bees naturally DWV-infected were collected from a Varroa mite-free apiary and divided into three experimental groups: naturally DWV infected bees, PBS injected bees, and artificially DWV super infected bees. Phenoloxidase activity and haemolymph cellular subtype count were investigated. Phenoloxidase activity was highest (P<0.05) in DWV-superinfected bees, and the haemocyte population differed within the three observed groups. Although, immune responses following DWV infection have still not been completely clarified, this investigation sheds light on the relation between cell immunity and the phenoloxidase activity of DWV-naturally infected honeybees exposed to additional stress such as injury and viral superinfection.(AU)

O sistema imune inato das abelhas consiste principalmente em peptídeos antimicrobianos, imunidade celular e melanização. Para investigar a resposta imune das abelhas a estressores imunológicos, foram testados três graus de estresse. Abelhas recém-emergidas naturalmente infectadas por DWV foram coletadas de um apiário livre de Varroa e divididas em três grupos experimentais: abelhas naturalmente infectadas por DWV, abelhas injetadas com PBS e abelhas superinfectadas artificialmente com DWV. A atividade de fenoloxidase e a contagem de subtipos celulares de hemolinfa foram investigadas. A atividade da fenoloxidase foi maior (P<0,05) nas abelhas super-infectadas com DWV, e a população de hemócitos diferiu entre os três grupos observados. Embora as respostas imunes após a infecção pelo DWV ainda não tenham sido completamente esclarecidas, esta investigação lança luz sobre a relação entre a imunidade celular e a atividade da fenoloxidase das abelhas infectadas naturalmente pelo DWV, expostas a estresse adicional, como lesão e superinfecção viral.(AU)

Evaluation of cell damage and modulation of cytokines TNF-, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy

Abstract 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.

Resumo 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.

Pathomorphological and immunohistochemical evaluation of unilateral progressive Mooren’s ulcer in a cat

Background: Mooren’s ulcer is a chronic and painful ulceration of the cornea. It begins progressively in the periphery and spread centrally in cornea. In human, it is seen uniaterally in most of cases. Mooren ulcer has not been reported in any kind of animals up to now. Although its aetiology is not completely enlighted, it has been suspected of the inflammatory reaction against injuries-microbiological and immun mediated effects. Immun response in presence of accumulation of immune complexes into the limbal vessels.As a result of the deficit in the regulatory mechanism because the number of suppressor cells control over B and T lymphocytes, These situations can result in a progressive tendancy to inflammation because the production of autoantibodies and/or lymphokine from cytotoxic T-lymhocytes creates an immune-mediated vasculitis. Numerous immigrant inflammatory cells and proteins are evaded from vessels. After triggering inflammatory cells and releasing of meditors, corneal vascularization, scar tissue and re-epithelization develop. This regenerative-reperative process plays an important role during post-inflammatory process.Case: In this case, it was aimed to detect pathomorphological structure and immunologic relations in progressive Mooren’s ulcer (MU). A 1 year-old mix breed cat was submitted to clinic with complaints of progressive painful and eyesight loss in left eye. There were 1 cm-ulceration, opacification and old haemorrhagic areas at peripheral cornea. Histopathologically, there was wide ulceration including all layers of corneal epithelium and particularly vacuolar degeneration at suprabasal cells. In corneal stroma, numerous neutrophiles and mononuclear cells were infiltrated. Neovascularisation and fibrosis beginning from limbus were also present. This fibrotic progress was confirmed by Masson’s trichrome staining method.[...]

Pathomorphological and immunohistochemical evaluation of unilateral progressive Moorens ulcer in a cat

Background: Moorens ulcer is a chronic and painful ulceration of the cornea. It begins progressively in the periphery and spread centrally in cornea. In human, it is seen uniaterally in most of cases. Mooren ulcer has not been reported in any kind of animals up to now. Although its aetiology is not completely enlighted, it has been suspected of the inflammatory reaction against injuries-microbiological and immun mediated effects. Immun response in presence of accumulation of immune complexes into the limbal vessels.As a result of the deficit in the regulatory mechanism because the number of suppressor cells control over B and T lymphocytes, These situations can result in a progressive tendancy to inflammation because the production of autoantibodies and/or lymphokine from cytotoxic T-lymhocytes creates an immune-mediated vasculitis. Numerous immigrant inflammatory cells and proteins are evaded from vessels. After triggering inflammatory cells and releasing of meditors, corneal vascularization, scar tissue and re-epithelization develop. This regenerative-reperative process plays an important role during post-inflammatory process.Case: In this case, it was aimed to detect pathomorphological structure and immunologic relations in progressive Moorens ulcer (MU). A 1 year-old mix breed cat was submitted to clinic with complaints of progressive painful and eyesight loss in left eye. There were 1 cm-ulceration, opacification and old haemorrhagic areas at peripheral cornea. Histopathologically, there was wide ulceration including all layers of corneal epithelium and particularly vacuolar degeneration at suprabasal cells. In corneal stroma, numerous neutrophiles and mononuclear cells were infiltrated. Neovascularisation and fibrosis beginning from limbus were also present. This fibrotic progress was confirmed by Massons trichrome staining method.[...](AU)

Immunotherapeutic potential of Crotoxin: anti-inflammatory and immunosuppressive properties

For the past 80 years, Crotoxin has become one of the most investigated isolated toxins from snake venoms, partially due to its major role as the main toxic component in the venom of the South American rattlesnake Crotalus durissus terrificus. However, in the past decades, progressive studies have led researchers to shift their focus on Crotoxin, opening novel perspectives and applications as a therapeutic approach. Although this toxin acts on a wide variety of biological events, the modulation of immune responses is considered as one of its most relevant behaviors. Therefore, the present review describes the scientific investigations on the capacity of Crotoxin to modulate anti-inflammatory and immunosuppressive responses, and its application as a medicinal immunopharmacological approach. In addition, this review will also discuss its mechanisms, involving cellular and molecular pathways, capable of improving pathological alterations related to immune-associated disorders.(AU)