Indoleamine-2,3 dioxygenase: a fate-changer of the tumor microenvironment.

Indoleamine-2,3 dioxygenase is a rate-limiting enzyme in the tryptophan catabolism in kynurenine pathways that has an immunosuppressive effect and supports cancer cells to evade the immune system in different cancer types. Diverse cytokines and pathways upregulate the production of indoleamine-2,3 dioxygenase enzymes in the tumor microenvironment and cause more production and activity of this enzyme. Ultimately, this situation results in anti-tumor immune suppression which is in favor of tumor growth. Several inhibitors such as 1-methyl-tryptophan have been introduced for indoleamine-2,3 dioxygenase enzyme and some of them are widely utilized in pre-clinical and clinical trials. Importantly at the molecular level, indoleamine-2,3 dioxygenase is positioned in a series of intricate signaling and molecular networks. Here, the main objective is to provide a focused view of indoleamine-2,3 dioxygenase enhancer pathways and propose further studies to cover the gap in available information on the function of indoleamine-2,3 dioxygenase enzyme in the tumor microenvironment.

IDO blockade negatively regulates the CTLA-4 signaling in breast cancer cells.

Cancer is classified into metabolic and/or genetic disorders; notably, the tryptophan catabolism pathway is vital in different cancer types. Here, we focused on the interaction and molecular connection between the cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) receptor and indoleamine-2,3-dioxygenase (IDO) enzyme. To test the impact of the selected immunotherapies on breast cancer cell migration and cell survival, we used in vitro assays. Also, we test the impact of anti-CTLA-4 antibody on the IDO-positive cells. The results of cell migration and clonogenic assays showed that anti-CTLA-4 antibody reduces cancer cell migration and clonogenic abilities of murine breast cancer cells. In addition, the result of flow cytometry showed that the anti-CTLA-4 antibody did not change the percentage of IDO-positive cancer cells. Notably, administrating an IDO blocker, 1-Methyl-DL-tryptophan (1MT), reduces the efficiency of the antiCTLA-4 antibody. The enzymatic blocking of the IDO reduces the efficiency of the anti-CTLA-4 antibody on cell migration and clonogenic abilities suggesting that there is an inhibitory interaction at the molecular level between functions of CTLA-4 and IDO. It is unclear via which mechanism(s) IDO interacts with CTLA-4 signaling and also why blocking IDO makes disruption in CTLA-4 signaling in cancer cells. Indeed, evaluating the role of IDO in CTLA-4 signaling in cancer cells may assist in clarifying a poor response to CTLA-4 immunotherapies by some patients. Hence, further investigation of the molecular interaction between CTLA-4 and IDO might help to improve the efficiency of CTLA-4 immunotherapy.

Association of Systemic Inflammatory and Immune Indices With Survival in Canine Patients With Oral Melanoma, Treated With Experimental Immunotherapy Alone or Experimental Immunotherapy Plus Metronomic Chemotherapy.

Analysis of the expression of inflammatory markers before starting treatment in human patients with cancer helps to predict outcomes and prognosis; however, there have been few studies on this topic in veterinary medicine. The present study aimed to evaluate inflammatory indices before treatment with autologous antitumor vaccine alone or this vaccine plus metronomic chemotherapy (MC) to predict response and prognosis. The indices included the neutrophil-lymphocyte ratio (NRL), platelet-lymphocyte ratio (PLR), monocyte-lymphocyte ratio (MLR), systemic immune-inflammation index (SII), C-reactive-protein-albumin ratio (CRP/ALB), lactate dehydrogenase level (LDH), frequency of blood lymphocyte subsets (CD4+, CD8+, Treg, and CD4/CD8 ratio) and frequency of blood myeloid-derived suppressor cells (MDSCs: monocytic [M]- MDSCs, and granulocytic [PMN]-MDSCs). Blood samples were collected from 25 dogs with oral melanoma treated with the autologous antitumor vaccine and from nine dogs that received MC plus vaccine before surgery. There were no statistically significant differences in the progression-free survival (PFS) or overall survival (OS) between the groups. In addition to the clinical stage, the CRP/ALB ratio and blood circulating Tregs in the univariate analysis showed an association with PFS and OS, and thus were selected for multivariable analysis. The CRP/ALB ratio was associated with PFS [hazard ratio (HR), 1.1; 95% confidence interval (CI), 1.0-1.1; p = 0.017] and OS [HR, 1; 95%CI, 1.0-1.1; p = 0.023]. Similarly, Treg was associated with PFS (HR, 1.6; 95% CI, 1.2-2.1; p = 0.001) and OS (HR, 1.6; 95% CI, 1.2-2.1; p = 0.001). Furthermore, canine patients with a CRP/ALB ratio above the cut-off point of 1.9 (established by receiver operating characteristic curve analysis) had worse PFS and OS, indicating the impact of the preoperative CRP/ALB ratio on the PFS and OS of dogs with oral melanoma. The CRP/ALB ratio and frequency of circulating Tregs are potential prognostic markers in dogs with oral melanoma.

Methylene blue and photodynamic therapy for melanomas: Inducing different rates of cell death (necrosis and apoptosis) in B16-F10 melanoma cells according to methylene blue concentration and energy dose.

BACKGROUND: Photodynamic therapy (PDT) is a cancer treatment based on the interaction between the photosensitizing agent methylene blue (MB), light, and molecular oxygen. MB has antibacterial properties and can to bind to melanin. Here, we investigated whether MB based PDT (MB-PDT) could decrease viability and induce death of murine melanoma B16-F10 cells. METHODS: B16-F10 cells were incubated with different concentrations of MB (0, 1, or 2 µg/mL) and exposed to a diode red laser with a wavelength of 660 nm and power output of 100 mW/cm2. The energy dose and density varied from 0 J and 0 J/cm2 to 100.8 J and 720 J/cm². Cell viability was measured using the trypan blue exclusion assay of cell viability and confirmed by performing an MTT assay. The morphological type and cell death rates were determined using fluorescence microscopy with acridine orange and ethidium bromide. The presence and rate of apoptosis were evaluated via Annexin V-Alexa Fluor/propidium iodide staining and flow cytometry analysis. RESULTS: MB-PDT decreased cell viability and increased cell death (necrosis and apoptosis) in a drug- and light-dose dependent manner. Morphological characteristics of necrosis were observed immediately after treatment, and apoptotic characteristics were observed after 3 h. The apoptosis and necrosis rates varied with the drug and light doses, with 2 µg/mL MB and a 100.8 J energy dose inducing the highest rates. CONCLUSIONS: We demonstrated that MB-PDT reduced murine melanoma B16-F10 cell viability and induced cell death in a drug- and light-dose dependent manner.

Quantification of Global DNA Methylation in Canine Melanotic and Amelanotic Oral Mucosal Melanomas and Peripheral Blood Leukocytes From the Same Patients With OMM: First Study.

Oral mucosal melanomas (OMMs) are aggressive and resistant cancers of high importance in veterinary oncology. Amelanotic OMM produces comparatively less melanin and is considered to be more aggressive than melanotic OMM. Global DNA methylation profiles with hypomethylated or hypermethylated patterns have both been associated with aggressive neoplasms; however, global DNA hypomethylation seems to correlate to higher aggressiveness. Accordingly, global DNA methylation in peripheral blood leukocytes has been investigated to understand the role of systemic or environmental factors in cancer development. This study aimed to quantify global DNA methylation in canine melanotic and amelanotic OMM samples and in the peripheral blood leukocytes of the same dogs. Tumor tissue samples were collected from 38 dogs, of which 19 were melanotic and 19 were amelanotic OMM. These were submitted to immunohistochemistry (IHC) with anti-5-methylcytosine (5mC) and anti-Ki67 primary antibodies. Ki67- and 5mC-positive nuclei were manually scored with the help of an image analysis system. Peripheral blood samples were collected from 18 among the 38 OMM-bearing dogs and from 7 additional healthy control dogs. Peripheral blood leukocytes were isolated from the 25 dogs, and DNA was extracted and analyzed by high-performance liquid chromatography (HPLC) for global DNA methylation. The pattern of global DNA methylation in both canine melanotic and amelanotic OMM indicated higher percentages of weakly or negatively stained nuclei in most of the OMM cells, presuming predominant global DNA hypomethylation. In addition, Ki67 counts in amelanotic OMM were significantly higher than those in melanotic OMM (p < 0.001). Global DNA methylation different immunostaining patterns (strong, weak or negative) correlated with Ki67 scores. Global DNA methylation in circulating leukocytes did not differ between the 9 melanotic and 9 amelanotic OMM or between the 18 OMM-bearing dogs and the 7 healthy dogs. This study provides new information on canine melanotic and amelanotic OMM based on global DNA methylation and cell proliferation.

Intratumoral (Poly-ICLC) Therapy for Dogs with Advanced Cancers: First Report on Clinical Effectiveness, Quality of Life, and Adverse Events.

Polyinosinic-polycytidylic acid-poly-l-lysine carboxymethylcellulose (poly-ICLC) is a synthetic double-stranded viral RNA analog widely tested as a component of human therapeutic cancer vaccines and as a standalone agent for treating human cancers. However, there are no reports on the use of poly-ICLC for treating canine cancers. This study aimed to investigate the clinical efficacy, quality of life (QL), and adverse events of poly-ICLC treatment in dogs with advanced cancers. The treatment protocol consisted of weekly intratumoral doses of poly-ICLC. The canine patients underwent clinical, laboratory, and imaging tests, and their owners answered weekly QL questionnaires. Fourteen canine patients with different types of spontaneous advanced tumors were enrolled. Most dogs had received prior conventional therapies. Five dogs received at least 12 doses of poly-ICLC: the injected tumor was stable in three dogs, there was a partial response in one, and the injected tumor significantly enlarged in the other. The QL scoring remained stable or increased in most cases. Mild adverse events related to poly-ICLC were observed in 10 of the 14 patients. The data showed that intratumoral poly-ICLC therapy was well tolerated in dogs with advanced cancers, with clinical benefit and improved QL scores observed in some dogs.

Inhibitory Effects of a Reengineered Anthrax Toxin on Canine and Human Osteosarcoma Cells.

Canine and human osteosarcomas (OSA) share similarities. Novel therapies are necessary for these tumours. The Bacillus anthracis toxin was reengineered to target and kill cells with high expressions of matrix metalloproteinases (MMPs) and urokinase plasminogen activator (uPA). Since canine OSA express MMPs and uPA, we assessed whether the reengineered toxin could show efficacy against these tumours. Two OSA cell lines (canine D17 and human MG63) and a non-neoplastic canine osteoblastic cell line (COBS) were used. Cells were treated with different concentrations of the reengineered anthrax toxin and cell viability was quantified using MTT assay. The cell cycle, apoptosis, and necrosis were analysed by flow cytometry. The wound-healing assay was performed to quantify the migration capacity of treated cells. D17 and MG63 cells had significantly decreased viability after 24 h of treatment. Cell cycle analysis revealed that OSA cells underwent apoptosis when treated with the toxin, whereas COBS cells arrested in the G1 phase. The wound-healing assay showed that D17 and MG63 cells had a significantly reduced migration capacity after treatment. These results point for the first time towards the in vitro inhibitory effects of the reengineered anthrax toxin on OSA cells; this reengineered toxin could be further tested as a new therapy for OSA.

Gene expression of the immunoinflammatory and immunological status of obese dogs before and after weight loss.

Obesity is characterized by a low degree of chronic inflammation state that, along with metabolic modifications, promotes important changes in the animal's organism. Adipose tissue actively participates in inflammation and immunity, and several defense cells of the organism may, therefore, be involved in the diversity found between obese and ideal weight individuals. Studies regarding this subject have shown immune cell changes in humans and rats, however, the literature is scarce in relation to dogs. Thus, the present study aimed to evaluate the gene expression profile of immunoinflammatory response and the lymphoproliferation of obese dogs before and after weight loss. Eight female dogs, neutered, of different breeds, aged between 1 and 8 years (4.74±3.19), obese, with body condition score (BCS) of 9 out of a 9-point scale and body composition determined by the deuterium isotope dilution method were included. The obese dogs were enrolled in a weight loss program and after losing 20% of their initial weight became a second experimental group. A third experimental group consisted of eight female dogs, neutered, aged between 1 and 8 years (3.11±0.78) and with ideal BCS (5 out of a 9-point scale). Gene expression of immunoinflammatory cytokines (resistin, leptin, adiponectin, TNF-α, IL-6, IL-8, and IL-10) was assessed by qRT-PCR and immunity was assessed by lymphoproliferative response using the flow cytometry technique. The data that presented normal distribution was evaluated by analysis of variance by the PROC MIXED of the SAS and when differences were detected, these were compared by the Tukey test. Regarding the gene expression data, the procedure PROC GLIMMIX was adopted and the methodology of generalized linear model was used, in which the Gama distribution proved to be adequate. Values of p<0.05 were considered significant. The mean weight loss period of the animals included in the study was 194.25 ± 28.31 days and the mean weekly weight loss rate was 1.02 ± 0.82%. The average fat mass, both in percentage (P<0.001) and in kilograms (P = 0.012), was higher in the obese group (40.88%; 8.91kg), returning to normal and without difference between the control group (19.16%; 3.01kg) and after weight loss (22.10%; 4.11kg). The weight loss program resulted in an increase in percentage of lean body mass (P = 0.001), 55.50% in obese animals vs 77.90% in obese dogs after weight loss, the latter with no difference when compared to the control group (80.84%). The obese group presented increased gene expression of resistin and IL-8 in relation to the weight loss group (P = 0.002). In adiponectin, the obese group presented increased mRNA gene expression when compared to the weight loss group (P = 0.003). The evaluation of lymphocyte proliferation showed differences between the group of obese animals before and after weight loss (P = 0.004). Weight loss resulted in an increase in the lymphoproliferation rate (18.48%) compared to obese dogs at the beginning of the study (10.71%). These results indicate that weight loss modulates the immunoinflammatory response of obese dogs and may present important benefits to health and longevity of dogs.

Functional characterization of neotropical snakes peripheral blood leukocytes subsets: linking flow cytometry cell features, microscopy images and serum corticosterone levels

Reptiles are the unique ectothermic amniotes, providing the key link between ectothermic anamniotes fish and amphibians, and endothermic birds and mammals; becoming an important group to study with the aim of providing significant knowledge into the evolutionary history of vertebrate immunity. Classification systems for reptiles' leukocytes have been described by their appearance rather than function, being still inconsistent. With the advent of modern techniques and the establishment of analytical protocols for snakes' blood by flow cytometry, we bring a qualitative and quantitative assessment of innate activities presented by snakes’ peripheral blood leukocytes, thereby linking flow cytometric features with fluorescent and light microscopy images. Moreover, since corticosterone is an important immunomodulator in reptiles, hormone levels of all blood samples were measured. We provide novel and additional information which should contribute to better understanding of the development of the immune system of reptiles and vertebrates.

Modulation of Cytokines Production by Indomethacin Acute Dose during the Evolution of Ehrlich Ascites Tumor in Mice.

The aim of the present study was to investigate the influence of a nonselective COX1/COX2 inhibitor (indomethacin) on tumor growth of Ehrlich Ascites Tumor (EAT) in mice, using as parameters the tumor growth and cytokine profile. Mice were inoculated with EAT cells and treated with indomethacin. After 1, 3, 6, 10, and 13 days the animals were evaluated for the secretion of TNFα, IL-1α, IL-2, IL-4, IL-6, IL-10, and IL-13 and PGE2 level in peritoneal cavity. The results have shown that EAT induces PGE2 production and increases tumor cells number from the 10th day. The cytokine profile showed EAT induces production of IL-6 from 10th day and of IL-2 on 13th day; the other studied cytokines were not affected in a significant way. The indomethacin treatment of EAT-bearing mice inhibited the tumor growth and PGE2 synthesis from the 10th day. In addition, the treatment of EAT-bearing mice with indomethacin has stimulated the IL-13 production and has significantly inhibited IL-6 in the 13th day of tumor growth. Taken together, the results have demonstrated that EAT growth is modulated by PGE2 and the inhibition of the tumor growth could be partly related to suppression of IL-6 and induction of IL-13.

An experimental study to compare inflammatory response due to liquid or gas joint distension in horses submitted to arthroscopy.

PURPOSE: To assess comparatively the inflammatory response that follows CO(2) or Ringer's lactate joint capsular distension of horses submitted to experimental arthroscopy METHODS: Each animal was submitted to a bilateral tarsocrural arthroscopy employing gas distention in one joint and fluid distention in the contralateral joint. Synovial fluid was evaluated at 0, six, 12, 24 and 48 hours post-operative. RESULTS: The use of CO(2) for arthroscopy causes an acute and mild synovitis alike to the liquid capsular distension, showing similar synovial fluid increase of leukocytes, TP, and TNF-α. Although synovial fluid PGE(2) content was higher in joints submitted to CO(2) distension, lower levels of hemoglobin and leukocytes oxidative burst after surgery indicates that CO(2) arthroscopy decreased intra-articular bleeding and activation of infiltrating leukocytes. CONCLUSIONS: The use of CO(2) for arthroscopic examination causes acute and mild synovitis that is similar to the effects caused by the liquid capsular distension. CO2 also seems to decrease intra-articular bleeding and activation of leukocytes.

An experimental study to compare inflammatory response due to liquid or gas joint distension in horses submitted to arthroscopy / Estudo experimental para comparar a resposta inflamatória decorrente da distensão líquida ou gasosa em cavalos submetidos à artroscopia

PURPOSE: To assess comparatively the inflammatory response that follows CO2 or Ringer's lactate joint capsular distension of horses submitted to experimental arthroscopy METHODS: Each animal was submitted to a bilateral tarsocrural arthroscopy employing gas distention in one joint and fluid distention in the contralateral joint. Synovial fluid was evaluated at 0, six, 12, 24 and 48 hours post-operative. RESULTS: The use of CO2 for arthroscopy causes an acute and mild synovitis alike to the liquid capsular distension, showing similar synovial fluid increase of leukocytes, TP, and TNF-α. Although synovial fluid PGE2 content was higher in joints submitted to CO2 distension, lower levels of hemoglobin and leukocytes oxidative burst after surgery indicates that CO2 arthroscopy decreased intra-articular bleeding and activation of infiltrating leukocytes. CONCLUSIONS: The use of CO2 for arthroscopic examination causes acute and mild synovitis that is similar to the effects caused by the liquid capsular distension. CO2 also seems to decrease intra-articular bleeding and activation of leukocytes.

OBJETIVO: Avaliar comparativamente a resposta inflamatória decorrente da distensão líquida ou gasosa em cavalos submetidos ao exame artroscópico. MÉTODOS: Cada animal foi submetido a uma artroscopia bilateral tarsocrural empregando uma distensão com gás em uma articulação e líquido na articulação contralateral. O líquido sinovial foi avaliado as zero, seis, 12, 24 e 48 horas do pós-operatório. RESULTADOS: A utilização de CO2 para a artroscopia provoca uma sinovite aguda e leve tal como a distensão capsular por líquido, mostrando um aumento similar de leucócitos, TP (proteína total) e TNF-a. Embora no líquido sinovial a quantidade de PGE2 tenha sido maior nas articulações submetidas à distensão por CO2, níveis mais baixos de hemoglobina e explosão oxidativa de leucócitos após a cirurgia indica que a artroscopia com CO2 diminuiu o sangramento intra-articular e ativação de leucócitos. CONCLUSÕES: A utilização de CO2 para exame artroscópico provoca uma sinovite aguda e leve que são semelhantes aos efeitos causados pela distensão capsular por líquido. O CO2 também parece diminuir o sangramento intra-articular e a ativação de leucócitos.

Metabolismo oxidativo de leucócitos em animais infectados pelo Vírus da Leucemia Bovina / Oxidative burst of leukocytes from bovine leukemia Virus-Infected Cattle

A infecção pelo vírus da leucemia bovina (VLB) leva ao desenvolvimento de linfocitose persistente (LP) ou linfossarcomas, principalmente em rebanhos bovinos leiteiros. Entretanto, os eventos que induzem tais manifestações ou o efeito da infecção na função das diferentes populações de leucócitos são pouco conhecidos. Avaliou-se o efeito da infecção pelo VLB na produção intracelular de peróxido de hidrogênio (H2O2) em leucócitos circulantes, mensurada pela fluorescência produzida pela diclorodihidrofluoresceína, utilizando-se de citometria de fluxo. As células foram obtidas de cinco vacas soronegativas; cinco vacas infectadas pelo VLB, alinfocitóticas; e cinco vacas infectadas, manifestando LP. Verificou-se que a infecção pelo VLB não altera a porcentagem de leucócitos circulantes produzindo H2O2, com ou sem prévio estímulo por adição in vitro: de 12-miristato 13-acetato de forbol (PMA); de lipopolissacarídeos de Escherichia coli (LPS); ou Staphylococcus aureus. Todavia, a produção de H2O2 em leucócitos de animais apresentando LP, com ou sem estímulo, foi menor que aquela verificada em leucócitos de animais soronegativos e de animais soropositivos alinfocitóticos. Os estímulos aumentaram a porcentagem de leucócitos produzindo H2O2 e a produção intracelular média em animais soronegativos ou naqueles infectados alinfocitóticos. Contudo, em leucócitos de vacas soropositivas manifestando LP, a fagocitose de S. aureus não elevou a porcentagem de leucócitos produzindo H2O2. Também, apenas a adição de PMA elevou a produção intracelular de H2O2 em leucócitos de fêmeas soropositivas manifestando LP. Concluiu-se que bovinos infectados pelo VLB, manifestando LP, apresentam menor produção intracelular de H2O2, demonstrando vulnerabilidade funcional refletida por imunossupressão.

Widespread Bovine Leukemia Virus (BLV) infection leads to persistent lymphocytosis (PL) or lymphosarcoma, mainly in dairy herds. Nevertheless, neither the sequence of events that conducts to these symptoms, nor the effect of infection on function of different leukocyte populations, is well known. We evaluated the effect of BLV infection on immune response of cows through the analysis of hydrogen peroxide (H2O2) intracellular production of circulating leukocytes after in vitro stimuli with phorbol-12-myristate-13-acetate (PMA), Escherichia coli lipopolysaccharides (LPS), or Staphylococcus aureus. Cells were obtained from five BLV-non infected cows, five BLV-infected, non-lymphocytotic cows, and five BLV-infected cows with PL, and analyzed by flow cytometry. Intracellular production of H2O2 was measured by dichlorodihydrofluorescein diacetate dependent fluorescence. Results showed that BLV infection does not alter the percentage of H2O2-producing circulating leukocytes (H2O2-PCL), with or without previous stimuli. However, the average of H2O2 intracellular production, with or without previous stimuli, in leukocytes obtained from cows with PL was smaller than those from cells obtained from BLV-negative cows and from BLV-infected, non-lymphocytotic cows. Moreover, stimuli increased H2O2 intracellular production and the percentage of H2O2-PCL obtained from BLVnegative cows and from BLV-infected, non-lymphocytotic cows. Conversely, neither phagocytosis of S. aureus and stimulus with LPS increases H2O2 intracellular production, nor phagocytosis increases the percentage of H2O2-PCL, when leukocytes were obtained from cows with PL. Thus, results show that BLV-infected cattle, with PL, have an impaired H2O2 intracellular production, demonstrating functional vulnerability reflected by immunosuppression.

Nepeta cataria L. var. citriodora (Becker) increases penile erection in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Nepeta cataria (NC), catnip, induces pleasure in cats and humans. AIM OF THE STUDY: Because sexual behavior is involved in pleasure, the effect of NC on sexual behavior and penile erection was evaluated in male rats that were acutely fed chow enriched with 10% NC leaves. Further, yawning was monitored because we previously demonstrated that NC modifies dopaminergic-related behaviors and that sexual behavior is closely linked with the dopaminergic system. The general activity and the motor coordination were examined to investigate the possible motor and emotional interferences of the sexual performance. MATERIAL AND METHODS: Male rats of the NC group received for a 4h period the chow enriched with 10% NC leaves while the control groups received regular chow. Fifteen min after the end of the 4h period of NC feeding the sexual behavior, apomorphine-induced penile erection and motor coordination were observed; the general activity in the open field was assessed 0, 15, 30 and 60 min after treatment. RESULTS: NC treatment increased male rat's penile erection. A slightly facilitation on male rat sexual behavior and a decreased in general activity of NC treated rats were observed. No effects on motor coordination and yawning episodes were detected by the NC treatment. CONCLUSIONS: It was suggested that NC increases penile erection and slightly improves male rat sexual behavior by an action on dopaminergic systems.

Assessing the impact of lung hyperinflation maneuver on systemic inflammatory response and lung collapse in patients undergoing surgeries under spontaneous ventilation.

BACKGROUND AND OBJECTIVES: Lung hyperinflation maneuvers (LHM) reverse intraoperative atelectasis; however, they can lead to pulmonary-induced systemic inflammatory response. The objective of this study was to determine the impact of LHM on systemic inflammatory response and lung structure in patients undergoing subarachnoid block. METHODS: After approval by the Ethics Committee of the institution and signing the informed consent, 20 patients undergoing small and medium surgical procedures were randomly separated into two groups: 1) control (CG), and 2) LHM (LHMG). One hour after the spinal anesthesia, LHM was performed in LHMG by applying bilevel positive pressure in the airways (BIPAP) with an expiratory pressure of 20 cmH(2)O and inspiratory pressure of 20 cmH(2)O for 1 to 2 minutes. Blood levels of TNFalpha, IL-1, IL-6, IL-8, IL-10, and IL-12 were determined by flow cytometry at baseline and at 90, 180, and 780 minutes. Lung volumes and weights were determined using CT scans obtained immediately after the surgery. RESULTS: The use of LHM resulted in a reduction in the fraction of non-aerated pulmonary parenchyma (7.5 +/- 4.3%, in the Control Group, vs. 4 +/- 2.1%, in the LHM Group, p = 0.02) without changing pulmonary volumes. A progressive increase in plasma levels of IL-1, IL-6, IL-8, and IL-10, similar in both groups, was observed. Plasma levels of TNFalpha and IL-12 were undetectable during the study. CONCLUSIONS: The use of LHM reduced the incidence of atelectasis, but it did not amplify the inflammatory response in patients with normal lungs undergoing small and medium surgeries under subarachnoid block.

Avaliação do impacto da aplicação de manobra de hiperinsuflação pulmonar sobre a resposta inflamatória sistêmica e colapso pulmonar em pacientes submetidos a procedimentos cirúrgicos sob ventilação espontânea / Assessing the impact of lung hyperinflation maneuver on systemic inflammatory response and lung collapse in patients undergoing surgeries under spontaneous ventilation

JUSTIFICATIVA E OBJETIVOS: O uso de manobras de hiperinsuflação pulmonar (MHP) reverte atelectasias intraoperatórias. Contudo, pode induzir resposta inflamatória sistêmica de origem pulmonar. O objetivo deste estudo foi testar o impacto da aplicação de MHP sobre a resposta inflamatória sistêmica e sobre a estrutura pulmonar em pacientes submetidos à anestesia subaracnoidea. MÉTODO: Após aprovação do Comitê de Ética institucional e obtenção do consentimento escrito pós-informado, 20 pacientes submetidos a procedimentos cirúrgicos de pequeno e médio porte foram alea torizados em dois grupos: 1) Controle (GC) e 2) MHP (GMHP). Uma hora após a instalação do bloqueio espinhal, foi realizada MHP no GMHP aplicando-se pressão positiva nas vias aéreas em dois níveis (BIPAP) com pressão expiratória de 20 cmH2O e pressão inspiratória de 20 cmH2O durante 1 a 2 minutos. TNFα, IL-1, IL-6, IL-8, IL-10 e IL-12 foram medidos no sangue através de técnica de citometria de fluxo nos momentos basal, 90, 180 e 780 minutos. Os volumes e peso pulmonares foram computados a partir de tomografias computadorizadas obtidas imediatamente após a cirurgia. RESULTADOS: A aplicação de MHP resultou em redução da fração de parênquima pulmonar não aerado (7,5 ± 4,3 por cento no Grupo Controle versus 4 ± 2,1 por cento no Grupo MHP, p = 0,02), sem alterações nos volumes pulmonares. Foi observada elevação progressiva nos valores plasmáticos das interleucinas IL-1, IL-6, IL-8 e IL-10, similar nos dois grupos. Os níveis plasmáticos de TNFα e IL-12 foram indetectáveis durante o estudo. CONCLUSÕES: A aplicação de MHP única reduziu a fração de atelectasias, porém não amplificou a resposta inflamatória observada em pacientes com pulmões normais submetidos a cirurgias de pequeno e médio portes sob anestesia subaracnoidea.

BACKGROUND AND OBJECTIVES: Lung hyperinflation maneuvers (LHM) reverse intraoperative atelectasis; however, they can lead to pulmonary-induced systemic inflammatory response. The objective of this study was to determine the impact of LHM on systemic inflammatory response and lung structure in patients undergoing subarachnoid block. METHODS: After approval by the Ethics Committee of the institution and signing the informed consent, 20 patients undergoing small and medium surgical procedures were randomly separated into two groups: 1) control (CG), and 2) LHM (LHMG). One hour after the spinal anesthesia, LHM was performed in LHMG by applying bilevel positive pressure in the airways (BIPAP) with an expiratory pressure of 20 cmH2O and inspiratory pressure of 20 cmH2O for 1 to 2 minutes. Blood levels of TNFα, IL-1, IL-6, IL-8, IL-10, and IL-12 were determined by flow cytometry at baseline and at 90, 180, and 780 minutes. Lung volumes and weights were determined using CT scans obtained immediately after the surgery. RESULTS: The use of LHM resulted in a reduction in the fraction of non-aerated pulmonary parenchyma (7.5 ± 4.3 percent, in the Control Group, vs. 4 ± 2.1 percent, in the LHM Group, p = 0.02) without changing pulmonary volumes. A progressive increase in plasma levels of IL-1, IL-6, IL-8, and IL-10, similar in both groups, was observed. Plasma levels of TNFα and IL-12 were undetectable during the study. CONCLUSIONS: The use of LHM reduced the incidence of atelectasis, but it did not amplify the inflammatory response in patients with normal lungs undergoing small and medium surgeries under subarachnoid block.

JUSTIFICATIVA Y OBJETIVOS: El uso de maniobras de hiperinsuflación pulmonar (MHP), revierte las atelectasias intraoperatorias, pero puede conllevar a una respuesta inflamatoria sistémica de origen pulmonar. Los objetivos de este estudio fueron comprobar el impacto de la aplicación de MHP sobre la respuesta inflamatoria sistémica y sobre la estructura pulmonar en pacientes sometidos a la anestesia subaracnoidea. MÉTODO: Después de la aprobación del Comité de Ética institucional y de la obtención del consentimiento informado, 20 pacientes sometidos a procedimientos quirúrgicos de pequeño y mediano porte, se separaron aleatoriamente en dos grupos: 1) control (GC) y 2) MHP (GMHP). Una hora después de la instalación del bloqueo espinal, fue realizada MHP en el GMHP aplicando una presión positiva en las vías aéreas en los niveles (BIPAP) con una presión espiratoria de 20 cmH2O y una presión inspiratoria de 20 cmH2O durante 1 a 2 minutos. FNTα, IL-1, IL-6, IL-8, IL-10 y IL-12 se midieron en la sangre a través de la técnica de citometría de flujo en los momentos basal, 90, 180 y 780 minutos. Los volúmenes y el peso pulmonares fueron computados a partir de tomografías computadorizadas obtenidas inmediatamente después de la operación. RESULTADOS: La aplicación de MHP resultó en una reducción de la fracción del parénquima pulmonar no aerado (7,5 ± 4,3 por ciento en el Grupo Control vs 4 ± 2,1 por ciento en el Grupo MHP, p = 0,02), sin alteraciones en los volúmenes pulmonares. Se observó una elevación progresiva en los valores plasmáticos de las interleucinas IL-1, IL-6, IL-8 y IL-10, similar en los dos grupos. Los niveles plasmáticos de FNT< y IL-12 no se detectaron durante el estudio. CONCLUSIONES: La aplicación de MHP única redujo la fracción de atelectasias, sin embargo, no amplificó la respuesta inflamatoria observada en pacientes con pulmones normales sometidos a cirugías de pequeño y mediano porte bajo anestesia subaracnoidea.

Influência da leucose enzoótica bovina na função fagocítica de leucócitos circulantes em animais manifestando linfocitose persistente / Influence of enzootic bovine leukosis on phagocytic function of circulating leukocytes from animals with persistent lymphocytosis

Avaliou-se, por citometria de fluxo, a fagocitose de Staphylococcus aureus conjugados com iodeto de propídio (IP), por leucócitos circulantes obtidos de cinco fêmeas bovinas negativas no sorodiagnóstico para a Leucose Enzoótica Bovina (LEB); de cinco fêmeas infectadas, manifestando linfocitose persistente (LP); e de cinco fêmeas infectadas, porém alinfocitóticas. Observou-se que, entre as amostras dos animais soronegativos, a porcentagem média de células realizando fagocitose (12,90%) não diferiu da observada entre as células dos animais alinfocitóticos (14,70%). Contudo, ambas foram maiores (p=0,047) que aquela verificada entre as células obtidas de animais manifestando LP (7,20%). Além disso, a intensidade média de fagocitose (caracterizada pela intensidade de fluorescência do IP, em valores arbitrários), verificada em leucócitos de animais manifestando LP (17,43) foi menor (p<0,001) que a observada em leucócitos de animais alinfocitóticos (29,50), e que a observada em leucócitos de animais soronegativos (25,18), que não diferiram entre si. Assim, os resultados permitem-nos alvitrar que há alteração na função fagocítica de leucócitos circulantes em animais infectados pelo vírus da LEB, manifestando LP.

This study evaluated the phagocytosis of propidium iodide-labeled Staphylococcus aureus (PI-Sa) by circulating leukocytes obtained from five Enzootic Bovine Leukosis (EBL)-negative cows, five naturally EBL-infected, non-lymphocytotic cows, and five EBL-positive cows with persistent lymphocytosis (PL), analyzed by flow cytometry. Among cells obtained from EBL-infected cows, presenting PL, the percentage of leukocytes carrying out phagocytosis (7.20%), was smaller (p=0.047) than that verified among cells obtained from non-infected (12.90%), and from BLV-infected, non-lymphocytotic cows (14.70%). Furthermore, leukocytes obtained from EBL-infected cows, presenting PL, showed smaller phagocytosis intensity (characterized by the intensity of propidium iodide fluorescence) than leukocytes obtained from non-infected and from EBL-infected, non-lymphocytotic, cows (p<0.001). Therefore, results show a decreased phagocytic function among circulating leukocytes obtained from BLV-infected, lymphocytotic cows.