BACKGROUND AND PURPOSE ATP is released in response to cellular damage, and P2X7 receptors have an essential role in the onset and maintenance of pathological changes. Haemorrhagic cystitis (HC) is a ...well‐known adverse effect of therapy with cyclophosphamide used for the treatment of many solid tumours and autoimmune conditions. Here we have evaluated the role of P2X7 receptors in a model of HC induced by cyclophosphamide.
EXPERIMENTAL APPROACH Effects of pharmacological antagonism or genetic deletion of P2X7 receptor on cyclophosphamide‐induced HC in mice was assessed by nociceptive and inflammatory measures. In addition, the presence of immunoreactive P2X7 receptors was assessed by immunohistochemistry.
KEY RESULTS Pretreatment with the selective P2X7 receptor antagonist A‐438079 or genetic ablation of P2X7 receptors reduced nociceptive behaviour scores in the HC model. The same strategies decreased both oedema and haemorrhage indices, on macroscopic or histological evaluation. Treatment with A‐438079 decreased the staining for c‐Fos in the lumbar spinal cord and brain cortical areas. Treatment with A‐438079 also prevented the increase of urinary bladder myeloperoxidase activity and macrophage migration induced by cyclophosphamide and reduced the tissue levels of IL‐1β and TNF‐α. Finally, P2X7 receptors were markedly up‐regulated in the bladders of mice with cyclophosphamide‐induced HC.
CONCLUSIONS AND IMPLICATIONS P2X7 receptors were significantly involved in a model of HC induced by cyclophosphamide. Pharmacological inhibition of these receptors might represent a new therapeutic option for this pathological condition.
Receptors of the P2X7 type have been demonstrated in granulocytes, monocytes/macrophages, B and T lymphocytes, and have been involved in several cellular mechanisms including those related to ...inflammation and immunological response. This study attempted to investigate the role of these receptors on the inflammatory and fibrogenic response in the kidneys of unilateral ureteral obstruction (UUO), by using P2X7 knockout mice (−/−). C57Bl6 mice were submitted to left UUO and killed after 7 and 14 days. Histopathology using hematoxylin–eosin, periodic-acid Schiff and Sirius-red staining, immunohistochemistry for macrophages, myofibroblasts, transforming growth factor-β (TGF-β)1 and P2X7, and immunofluorescence for apoptotic cells (terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick-end labeling) were performed. Protocols were as follows: (1) control; (2) sham; (3) control P2X7 (−/−); (4) sham P2X7 (−/−); (5) UUO wild type (WT); (6) UUO P2X7 (−/−). Myofibroblasts and Sirius-red staining were significantly lower in UUO P2X7 (−/−) mice at days 7 and 14, compared to UUO WT. Kidneys from UUO P2X7 (−/−) mice showed reduced number of inflammatory cells at day 14 but not at day 7, compared to UUO WT. TGF-β1 was less in UUO P2X7 (−/−) mice at days 7 and 14 when compared to UUO WT. Macrophage infiltration and tubular apoptosis were lower in UUO P2X7 (−/−) at day 14 but not at day 7, compared to UUO WT. P2X7 was expressed only in tubular epithelial cells at day 7 of UUO WT mice. These findings constitute the first evidence that P2X7 receptors are implicated in macrophage infiltration, collagen deposition and apoptosis in response to ureteral obstruction in mice.
Purpose
Incisional hernia (IH) occurs when there is a partial or complete solution of continuity of a fascia previously incised. Systematic reviews demonstrate that surgical treatment of IHs with the ...use of meshes are approximately 16%. Meta-analyses have demonstrated the superiority of mesh placement using sublay technique, but without a pathophysiological explanation. Thus, we aim to evaluate the different techniques of mesh positioning in an experimental model.
Methods
Fifty rats were distributed into five groups; control; simulation (SM)—submitted to laparotomy only; onlay—the mesh was positioned in onlay fashion; retromuscular (SL)—the mesh was positioned in a sublay fashion; intraperitoneal (IPOM)—positioning of the mesh adjacent to the transversalis fascia, inside the cavity. After 60 days, adhesions, tensiometry, histology, and immunohistochemistry were addressed.
Results
The IPOM group had the most adhesions, together with the SL group, with significantly relevant results. The SL group had higher values of tensiometric evaluation, while the IPOM group had the lowest mean in the tensiometry evaluation, being even lower than the SM group. Regarding histological and immunohistochemical findings, the SL group had a higher pixel number count compared to the groups, with statistical significance, in addition to higher expression of polymorphonuclear infiltrate and CD68 markers.
Conclusion
The mesh positioning in sublay compartment is associated with the development of more pronounce minimum tensile force required for detaching the surrounding abdominal wall tissues it was incorporated. The intensity of these findings correlates to the different histological and immunohistochemical profiles observed following each repair, since SL group was characterized by a higher proportion of collagen, inflammatory, and reparative elements. Characterizing these pro-healing elements and its counterparts will allow the development of new therapeutic tools which could be added to the still far-from-ideal current therapeutic options for IH treatment.
Chitosan is a biopolymer derived from chitin deacetylation, present in the exoskeleton of crustaceans and insects. Chitosan has been evaluated as rumen modulator and silage additive due to its ...antimicrobial properties. The objective of this study was to determine the effects of both chitosan and a bacterial additive on microbiological quality, chemical composition, nutrient in vitro degradation, fermentative profile, and total losses of whole-soybean plant silage (SS) harvested at R6 stage. Four treatments in a factorial arrangement were randomly assigned to 40 experimental minisilos as no additives (CON), 8 g/t fresh forage of microbial inoculant (INO; Kera SIL, Kera Nutrição Animal, Bento Gonçalves, Brazil); 5 g/kg of fresh forage chitosan (CHI); and CHI + INO. Microbial inoculant was composed of Lactobacillus plantarum (4.0 × 1010 cfu/g) and Propionibacterium acidipropionici (2.6 × 1010 cfu/g). The CHI and INO alone increased counts of lactic bacteria and anaerobic bacteria and decreased counts of mold and yeast in SS. The CHI or INO alone increased in vitro degradation of dry matter, crude protein, and neutral detergent fiber, and decreased nonfiber carbohydrate content of SS. Chitosan increased NH3-N and lactate concentrations and decreased ethanol concentration in SS. The CHI increased dry matter recovery from SS; INO increased silage aerobic stability. The combination of CHI+INO showed the lowest value of gas losses. In general, the combination of CHI and INO had small positive effects on gas losses of SS; however, both CHI or INO alone improved nutrient in vitro degradation and decreased mold and yeast in SS. Chitosan or INO utilization improves SS quality.
Lactation diets are composed mostly of carbohydrates that are not fully fermented by rumen microbes. The aim of this study was to evaluate exogenous fibrolytic (Fibrozyme, Alltech Inc., ...Nicholasville, KY) and amylolytic (Amaize, Alltech Inc.) enzymes on nutrient intake, sorting index, total-tract apparent digestibility, ruminal fermentation, nitrogen utilization, milk yield, and composition of dairy cows in mid-lactation. Thirty-two multiparous Holstein cows (181 ± 35 d in milk, 571 ± 72.7 kg of body weight, and 29.6 ± 5.24 kg/d of milk yield at the start of experiment) were blocked according to milk yield and randomly allocated to treatments in a 4 × 4 Latin square design. Treatments were (1) control, basal diet without exogenous enzymes; (2) fibrolytic enzyme (FIB), dietary supplementation of Fibrozyme at 12 g/d (51 IU of xylanase activity/kg of diet dry matter); (3) amylolytic enzyme (AMY), dietary supplementation of Amaize at 8 g/d (203 fungal amylase units/kg of diet dry matter); and (4) both fibrolytic and amylolytic enzymes (FIB+AMY) added at the same dose of the individual treatments. Enzyme products were added to the concentrate during its preparation (once a week). The supply of FIB and AMY had no effect on nutrient intake and digestibility. However, an interaction effect was observed on sorting index of feed particle size between 8 and 19 mm. Amylolytic enzyme increased the sorting for feed particles between 8 and 19 mm, only when fed without FIB. In addition, AMY decreased the sorting for feed with particle size greater than 19 mm. An interaction effect was observed between FIB and AMY for ruminal butyrate concentration and N excretion. Amylolytic enzyme increased ruminal butyrate concentration in cows treated with FIB. Further, FIB decreased milk protein production and feed efficiency only in cows not fed AMY. Amylolytic enzyme reduced urinary N excretion. Exogenous enzymes had no effect on milk production and composition of dairy cows. This study lacks evidence that fibrolytic and amylolytic enzymes can affect nutrient digestibility, ruminal fermentation, and performance of mid-lactation cows.
The goal of this study was to test the feasibility of BALB/c mice as an experimental model in the study of dengue disease. BALB/c mice were intraperitoneal infected with DENV-2 obtained from a human ...patient. Histopathological analysis of infected animals revealed liver injury with viral antigens detection. In initial stages, the most prominent lesions were vacuolization and diffuse steatosis in hepatocytes. Serum levels of ALT and AST increased progressively, reaching the highest values 7 days p.i. and decreasing at the 14th day. Since levels of circulating virus were very low, viremia was analyzed in C6/36 cells. Virus presence was detected by ultrastructural analysis, confirmed by RT-PCR assays. Period of viremia was analyzed by flow cytometry with cells incubated with mouse-infected sera collected in different days, revealing peak virus levels at the 7th day p.i. All such data correlate to the development of the disease described in humans.
The objective of the current study was to evaluate the effects of supplemental n-3 and n-6 fatty acid (FA) sources on cellular immune function of transition dairy cows. Animals were randomly assigned ...to receive 1 of 4 diets: control (n=11); whole flaxseed (n-3 FA source; n=11), 60 and 80g/kg of whole flaxseed diet dry matter (DM) basis during pre- and postpartum, respectively; whole raw soybeans (n-6 FA source; n=10), 120 and 160g/kg of whole raw soybeans (diet DM basis) during pre- and postpartum, respectively; and calcium salts of unsaturated FA (Megalac-E, n-6 FA source; n=10), 24 and 32g/kg of calcium salts of unsaturated FA (diet DM basis) during pre- and postpartum, respectively. Supplemental FA did not alter DM intake and milk yield but increased energy balance during the postpartum period. Diets containing n-3 and n-6 FA sources increased phagocytosis capacity of leukocytes and monocytes and phagocytosis activity of monocytes. Furthermore, n-3 FA source increased phagocytic capacity of leukocytes and neutrophils and increased phagocytic activity in monocytes and neutrophils when compared with n-6 FA sources. Supplemental FA effects on adaptive immune system included increased percentage of T-helper cells, T-cytotoxic cells, cells that expressed IL-2 receptors, and CD62 adhesion molecules. The results of this study suggest that unsaturated FA can modulate innate and adaptive cellular immunity and trigger a proinflammatory response. The n-3 FA seems to have a greater effect on phagocytic capacity and activity of leukocytes when compared with n-6 FA.
Galectin‐3 (gal‐3), a β‐galactoside‐binding animal lectin, plays a role in cell‐cell and cell‐extracellular matrix interactions. Extracellular gal‐3 modulates cell migration and adhesion in several ...physiological and pathological processes. Gal‐3 is highly expressed in activated macrophages. Schistosoma mansoni eggs display a large amount of gal‐3 ligands on their surface and elicit a well‐characterized, macrophage‐dependent, granulomatous, inflammatory reaction. Here, we have investigated the acute and chronic phases of S. mansoni infection in wild‐type and gal‐3−/− mice. In the absence of gal‐3, chronic‐phase granulomas were smaller in diameter, displaying thinner collagen fibers with a loose orientation. Schistosoma‐infected gal‐3−/− mice had remarkable changes in the monocyte/macrophage, eosinophil, and B lymphocyte subpopulations as compared with the infected wild‐type mice. We observed a reduction of macrophage number, an increase in eosinophil absolute number, and a decrease in B lymphocyte subpopulation (B220+/high cells) in the periphery during the evolution of the disease in gal‐3−/− mice. B lymphopenia was followed by an increase of plasma cell number in bone marrow, spleen, and mesenteric lymph nodes of the infected gal‐3−/− mice. The plasma IgG and IgE levels also increased in these mice. Gal‐3 plays a role in the organization, collagen distribution, and mobilization of inflammatory cells to chronic‐phase granulomas, niches for extramedullary myelopoiesis, besides interfering with monocyte‐to‐macrophage and B cell‐to‐plasma cell differentiation.
Objectives:
To evaluate somatostatin receptor 2A (SSTR2A) and dopamine receptor 2 (DR2) protein expression in somatotropinomas and to relate it to response to somatostatin analogues (SA).
Design and ...patients:
SSTR2A and DR2 expression was analyzed by immunohistochemistry in 88 somatotropinomas from patients submitted to either pre-surgical or adjuvant SA treatment. Tumors were scored according to percentage of immunostained cells: 0 (<25%), 1 (25–50%), and 2 (>50%). Relation between protein expression and response to SA was performed in 66 patients. Response to SA was assessed by percent IGF-I reduction, being considered as an IGF-I per cent reduction higher than 50%. Disease control was also assessed (GH<1.0 ng/ml and normal IGF-I).
Results:
SSTR2A and DR2 were expressed in 100% and 98% of tumors, respectively. Biochemical response and disease control rates were 48% and 32%, respectively. Median IGF-I percent reduction after 3 months of SA treatment was lower in the SSTR2A score 0 than in the scores 1 and 2 (
p
<0.001, both), and after 6 months in the score 0 than in the score 1 (
p
=0.001) and 2 (
p
<0.001). Biochemical response and disease control were associated with SSTR2 expression (
p
<0.001 and
p
=0.004, respectively). A negative predictive value for biochemical response of 100% was found when a SSTR2A expression <25% of immunostained cells cut-off point was considered. No relation was found between DR2 expression and biochemical response and disease control.
Conclusion:
SSTR2A and DR2 are highly expressed in somatotropinomas. Low SSTR2A, but not DR2, expression is a negative predictive factor to response to SA.
•Chitosan improved chemical composition, DM recovery, and NDF in vitro degradability of sugarcane silage.•All inoculants decreased ethanol concentration and pH from silage juice.•Silage treated with ...inoculants showed better aerobic stability than control.•Chitosan increased lactic-acid bacteria as compared to microbial inoculants.
The purpose of the current study was to evaluate the effects of inoculants on chemical composition, dry matter (DM) and neutral detergent fiber (aNDF) in vitro degradation, fermentative and effluent losses, microbiology, fermentative profile, and aerobic stability of sugarcane mini-silos. Treatments were randomly distributed to the mini-silos, in which: (1) Control (CON); (2) Lactobacillus buchneri (Lb), addition of Lb at 2.6×1010cfu/g; (3) Lactobacillus buchneri and Bacillus subtilis (Lb+Bs), addition of Lb at 2.6×1010cfu/g and Bs at 1×109cfu/g; and (4) Chitosan (CHI), addition of 1% of CHI on wet basis of sugarcane ensiled. Treatments 2 and 3 were incorporated to the silage at 2g/t of natural matter ensiled. Lb and Lb+Bs did not alter the in vitro degradation of DM and NDF. Chitosan incorporation increased the DM content (P=0.013, 18.7g/kg DM) and improved (P=0.029, 45.6g/kg DM) the NDF in vitro degradation of sugarcane silage. In addition, CHI incorporation showed higher (P=0.002) DM content in silage than Lb and Lb+Bs. Microbial inoculants (Lb and Lb+Bs) reduced the total losses (P=0.009) of sugarcane silage. Moreover, CHI incorporation showed lower (P=0.001, 84.9g/kg DM) total losses and higher (P=0.031, 84.8g/kg DM) dry matter recovery than Lb and Lb+Bs. Lactic acid bacteria concentration was increased (P=0.001) with additives, and CHI incorporation showed higher (P=0.001) lactic acid bacteria concentration than silages treated Lb and Lb+Bs. All additives decreased the ethanol concentration in sugarcane silage, but CHI showed lower (P=0.002) ethanol concentration compared to Lb and Lb+Bs. Inoculants improved the aerobic stability of sugarcane silage. In general, the incorporation of CHI to sugarcane silage showed better results of NDF in vitro degradation and gas and effluent losses than Lb and Lb+Bs. Moreover, CHI incorporation showed higher concentration of lactic acid bacteria and lower concentration of ethanol compared to silages treated Lb and Lb+Bs. Chitosan may be an alternative additive to microbial inoculants used in sugarcane ensiling.
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