Silencing of Sporothrix schenckii GP70 Reveals Its Contribution to Fungal Adhesion, Virulence, and the Host-Fungus Interaction.

Sporothrix schenckii is one of the etiological agents of sporotrichosis, a cutaneous and subcutaneous infection distributed worldwide. Like other medically relevant fungi, its cell wall is a molecular scaffold to display virulence factors, such as protective pigments, hydrolytic enzymes, and adhesins. Cell wall proteins with adhesive properties have been previously reported, but only a handful of them have been identified and characterized. One of them is Gp70, an abundant cell wall protein mainly found on the surface of yeast-like cells. Since the protein also has a role in the activity of 3-carboxy-cis,cis-muconate cyclase and its abundance is low in highly virulent strains, its role in the Sporothrix-host interaction remains unclear. Here, a set of GP70-silenced strains was generated, and the molecular and phenotypical characterization was performed. The results showed that mutants with high silencing levels showed a significant reduction in the adhesion to laminin and fibrinogen, enzyme activity, and defects in the cell wall composition, which included reduced mannose, rhamnose, and protein content, accompanied by an increment in ß-1,3-glucans levels. The cell wall N-linked glycan content was significantly reduced. These strains induced poor TNFα and IL-6 levels when interacting with human peripheral blood mononuclear cells in a dectin-1-, TLR2-, and TLR4-dependent stimulation. The IL-1ß and IL-10 levels were significantly higher and were stimulated via dectin-1. Phagocytosis and stimulation of neutrophil extracellular traps by human granulocytes were increased in highly GP70-silenced strains. Furthermore, these mutants showed virulence attenuation in the invertebrate model Galleria mellonella. Our results demonstrate that Gp70 is a versatile protein with adhesin properties, is responsible for the activity of 3-carboxy-cis,cis-muconate cyclase, and is relevant for the S. schenckii-host interaction.

The neonatal liver hosts a spontaneously occurring neutrophil population, exhibiting distinct spatial and functional characteristics from adults.

The elusive nature of the liver immune system in newborns remains an important challenge, casting a shadow over our understanding of how to effectively treat and prevent diseases in children. Therefore, deeper exploration into the intricacies of neonatal immunology might be crucial for improved pediatric healthcare. Using liver intravital microscopy, we unveiled a significant population of granulocytes in the hepatic parenchyma of fetuses and newborns. Utilizing high-dimensional immunophenotyping, we showed dynamic alterations predominantly in granulocytes during neonatal development. Liver intravital microscopy from birth through adulthood captures real-time dynamics, showing a substantial presence of Ly6G + cells that persisted significantly up to 2 weeks of age. Using CyTOF, we characterized neonatal Ly6G + cells as neutrophils, confirmed by morphology and immunohistochemistry. Surprisingly, the embryonic liver hosts a distinct population of neutrophils established as early as the second gestational week, challenging conventional notions about their origin. Additionally, we observed that embryonic neutrophils occupy preferentially the extravascular space, indicating their early establishment within the liver. Hepatic neutrophils in embryos and neonates form unique cell clusters, persisting during the initial days of life, while reduced migratory capabilities in neonates are observed, potentially compensating with increased reactive oxygen species (ROS) release in response to stimuli. Finally, in vivo imaging of acute neutrophil behavior in a newborn mouse, subjected to focal liver necrosis, unveils that neonatal neutrophils exhibit a reduced migratory response. The study provides unprecedented insights into the intricate interplay of neutrophils within the liver, shedding light on their functional and dynamic characteristics during development.

Inflammatory myopathies and beyond: The dual role of neutrophils in muscle damage and regeneration.

Skeletal muscle is one of the most abundant tissues of the human body and is responsible for the generation of movement. Muscle injuries can lead to severe disability. Skeletal muscle is characterized by an important regeneration capacity, which is possible due to the interaction between the myoblasts and immune cells. Neutrophils are fundamental as inducers of muscle damage and as promoters of the initial inflammatory response which eventually allows the muscle repair. The main functions of the neutrophils are phagocytosis, respiratory burst, degranulation, and the production of neutrophil extracellular traps (NETs). An overactivation of neutrophils after muscle injuries may lead to an expansion of the initial damage and can hamper the successful muscle repair. The importance of neutrophils as inducers of muscle damage extends beyond acute muscle injury and recently, neutrophils have become more relevant as part of the immunopathogenesis of chronic muscle diseases like idiopathic inflammatory myopathies (IIM). This heterogeneous group of systemic autoimmune diseases is characterized by the presence of muscle inflammation with a variable amount of extramuscular features. In IIM, neutrophils have been found to have a role as biomarkers of disease activity, and their expansion in peripheral blood is related to certain clinical features like interstitial lung disease (ILD) and cancer. On the other hand, low density granulocytes (LDG) are a distinctive subtype of neutrophils characterized by an enhanced production of NETs. These cells along with the NETs have also been related to disease activity and certain clinical features like ILD, vasculopathy, calcinosis, dermatosis, and cutaneous ulcers. The role of NETs in the immunopathogenesis of IIM is supported by an enhanced production and deficient degradation of NETs that have been observed in patients with dermatomyositis and anti-synthetase syndrome. Finally, new interest has arisen in the study of other phenotypes of LDG with a phenotype corresponding to myeloid-derived suppressor cells, which were also found to be expanded in patients with IIM and were related to disease activity. In this review, we discuss the role of neutrophils as both orchestrators of muscle repair and inducers of muscle damage, focusing on the immunopathogenesis of IIM.

Intestinal morphometry and cell density features in tropical farmed fish: A methodological approach.

The digestive tract of fish has many morphological adaptations related to habitat and nutrition. Intestinal biometry may reflect these adaptations. Here, we aimed to describe histometric patterns in farmed fish and their relationship with feeding by using a standardized protocol considering cell density by tissue area. Five juvenile specimens of each species (Pseudoplatystoma corruscans, Piaractus mesopotamicus, and Oreochromis niloticus) were used. O. niloticus possessed higher intestinal weight and length besides higher intestinal quotient and intestinal somatic index than the other species. The general histological composition was similar between species. However, P. corruscans showed differences in thickness between the anterior and posterior segments. O. niloticus had thinner serosa and muscularis layers than the other species. The cell density was distinct in both species and segments. Comparing the intestinal segments, O. niloticus displayed the lowest count of granulocytes. Goblet cell density was lower in P. mesopotamicus in all segments. However, the volume of these cells was higher in the anterior and middle anterior segments. Our data demonstrated that intestinal structural plasticity is associated with the difference in feeding habits. Here, we used quantitative standardized histometric criteria to understand the morphophysiological diversity of the fish digestive tract, and this technique can be applied in future studies to evaluate changes in the digestive tracts of vertebrates.

Usefulness of Immature Granulocytes as A Prognostic Factor in ST-Elevation Myocardial Infarction

ABSTRACT Objective: ST-segment elevation myocardial infarction (STEMI) is a serious, life-threatening disease. Inflammatory markers have recently become the focus of attention in the assessment of severity in the early stages of STEMI. This study aimed to evaluate the importance of immature granulocytes (IG) as a prognostic marker in STEMI. Methods: Patients admitted to the coronary care unit with a diagnosis of STEMI and who underwent primary percutaneous coronary intervention (pPCI) within the period from January 1, 2019 to January 1, 2020, were retrospectively scanned. A total of 146 patients were analised; of these, 112 (76.7%) were male and 34 (33.3) were female, with a mean age of 62.65±14.06 years. Patients' age, gender, haemogram, biochemistry, and mortality results were recorded. The patients were divided into two groups as low (<0.6) and high (≥0.6) IG levels and compared. Results: The mean IG levels were significantly higher in the non-survivor group compared to the survivor group (1.12±0.22 vs. 0.50±0.28, P<0.001). Mortality rates were significantly higher in the high IG group compared to the low IG group (26.9% vs. 9.6%, P=0.006). IG was shown to predict mortality with a sensitivity of 72.2% and a specificity of 77.8% at a cut-off value of 0.65 (area under the curve: 0.740, 95% CI: 0.635-0.846, P<0.001). Conclusion: High IG values in the blood collected at the time of admission to the emergency department are a marker of mortality in patients with STEMI.

Neutrophil depletion in the pre-implantation phase impairs pregnancy index, placenta and fetus development.

Maternal neutrophils cells are players in gestational tolerance and fetus delivery. Nonetheless, their actions in each phase of the pregnancy are unknown. We here investigated the role of maternal neutrophil depletion before the blastocyst implantation phase and outcomes in the pregnancy index, placenta, and fetus development. Neutrophils were pharmacologically depleted by i.p. injection of anti-Gr1 (anti-neutrophils; 200 µg) 24 hours after plug visualization in allogeneic-mated C57BL/6/BALB/c mice. Depletion of peripheral neutrophils lasted until 48 hours after anti-Gr1 injection (gestational day 1.5-3.5). On gestational day 5.5, neutrophil depletion impaired the blastocyst implantation, as 50% of pregnant mice presented reduced implantation sites. On gestational day 18.5, neutrophil depletion reduced the pregnancy rate and index, altered the placenta disposition in the uterine horns, and modified the structure of the placenta, detected by reduced junctional zone, associated with decreased numbers of giant trophoblast cells, spongiotrophoblast. Reduced number of placenta cells labeled for vascular endothelial growth factor (VEGF), platelet-endothelial cell adhesion molecule (PECAM-1), and intercellular cell adhesion molecule (ICAM-1), important markers of angiogenesis and adhesiveness, were detected in neutrophil depleted mice. Furthermore, neutrophil depletion promoted a higher frequency of monocytes, natural killers, and T regulatory cells, and lower frequency of cytotoxic T cells in the blood, and abnormal development of offspring. Associated data obtained herein highlight the pivotal role of neutrophils actions in the early stages of pregnancy, and address further investigations on the imbricating signaling evoked by neutrophils in the trophoblastic interaction with uterine epithelium.

CD3Ɛ immune restorative ability induced by Maitake Pro4x in immunosupressed BALBc mice.

OBJECTIVES: The aim of this research was to determine if the rich beta glucan compound called Maitake Pro4X can recover the T cell/NK population depleted by Dexamethasone treatment in lymph nodes from cancer-free BALBc female mice. A CD3Ɛ molecular FITC labelled marker was used to measure the effect of Maitake D-Fraction Pro4X (5 mg/kg) on T cell/NK cells populations employing flow cytometry from immunosuppressed female BALBc mice in lymph nodes. There were employed other molecular markers such as CD19, CD105, Ly6G. RESULTS: Maitake Pro4X (5 mg/kg) was able to recover 42.97% of the depleted CD3Ɛ FITC cell population level in Lymph nodes from immunosuppressed female BALBc mice from 4.328 ± 6.229 to 22.646 ± 12.393 (p < 0.01) using Flow Cytometry. Maitake was also able to significantly increase the Ly6G PE cell population with p < 0.05 in lymph nodes.

The interplay between serine proteases and caspase-1 regulates the autophagy-mediated secretion of Interleukin-1 beta in human neutrophils.

Neutrophils play major roles against bacteria and fungi infections not only due to their microbicide properties but also because they release mediators like Interleukin-1 beta (IL-1ß) that contribute to orchestrate the inflammatory response. This cytokine is a leaderless protein synthesized in the cytoplasm as a precursor (pro-IL-1ß) that is proteolytically processed to its active isoform and released from human neutrophils by secretory autophagy. In most myeloid cells, pro-IL-1ß is processed by caspase-1 upon inflammasome activation. Here we employed neutrophils from both healthy donors and patients with a gain-of-function (GOF) NLRP3-mutation to dissect IL-1ß processing in these cells. We found that although caspase-1 is required for IL-1ß secretion, it undergoes rapid inactivation, and instead, neutrophil serine proteases play a key role in pro-IL-1ß processing. Our findings bring to light distinctive features of the regulation of caspase-1 activity in human neutrophils and reveal new molecular mechanisms that control human neutrophil IL-1ß secretion.

Echocardiographic epicardial fat thickness and immature granulocyte are novel inflammatory predictors of acute ischemic stroke: a prospective study

ABSTRACT BACKGROUND: Acute ischemic stroke (AIS) is the most common type of stroke. Inflammation is the primary factor in the pathogenesis of atherosclerosis. Use of immature granulocytes (IGs) has been recommended as a new indicator of systemic inflammation. However, data on the association between echocardiographic epicardial fat tissue thickness (EFT) and IGs in patients with AIS are limited. OBJECTIVE: To evaluate the association between the presences of IGs, epicardial fat tissue and AIS. DESIGN AND SETTING: Prospective study in a tertiary-care university hospital in Antalya, Turkey. METHODS: Our study included 53 AIS patients and 41 healthy controls with age and gender compatibility. Blood samples and transthoracic echocardiography of all participants were compared. RESULTS: IG levels were significantly higher in patients with AIS than in controls (0.62 ± 0.36 versus 0.28 ± 0.02, P < 0.001). The mean EFT was 3.74 ± 0.61 mm in the control group and 6.33 ± 1.47 mm in the AIS patient group. EFT was significantly greater in AIS patients than in controls (P < 0.001). For the optimum cut-off value for IG (0.95), the area under the curve (AUC) was determined to be 0.840; sensitivity was determined to be 81.1% and specificity, 92.5%. For the optimum cut-off value for EFT (4.95 mm), the AUC was determined to be 0.953; sensitivity was determined to be 90.6% and specificity, 90%. CONCLUSIONS: IG and echocardiographic EFT are clinical markers that can be used to predict AIS risk.

Usefulness of Immature Granulocytes as A Prognostic Factor in ST-Elevation Myocardial Infarction.

OBJECTIVE: ST-segment elevation myocardial infarction (STEMI) is a serious, life-threatening disease. Inflammatory markers have recently become the focus of attention in the assessment of severity in the early stages of STEMI. This study aimed to evaluate the importance of immature granulocytes (IG) as a prognostic marker in STEMI. METHODS: Patients admitted to the coronary care unit with a diagnosis of STEMI and who underwent primary percutaneous coronary intervention (pPCI) within the period from January 1, 2019 to January 1, 2020, were retrospectively scanned. A total of 146 patients were analised; of these, 112 (76.7%) were male and 34 (33.3) were female, with a mean age of 62.65±14.06 years. Patients' age, gender, haemogram, biochemistry, and mortality results were recorded. The patients were divided into two groups as low (<0.6) and high (≥0.6) IG levels and compared. RESULTS: The mean IG levels were significantly higher in the non-survivor group compared to the survivor group (1.12±0.22 vs. 0.50±0.28, P<0.001). Mortality rates were significantly higher in the high IG group compared to the low IG group (26.9% vs. 9.6%, P=0.006). IG was shown to predict mortality with a sensitivity of 72.2% and a specificity of 77.8% at a cut-off value of 0.65 (area under the curve: 0.740, 95% CI: 0.635-0.846, P<0.001). CONCLUSION: High IG values in the blood collected at the time of admission to the emergency department are a marker of mortality in patients with STEMI.

Automated Versus Manual Band Counts for the Diagnosis of Invasive Bacterial Infections in Infants Who Are Febrile.

We conducted a secondary analysis of a prospective study of infants ≤60 days of age who were febrile to assess the diagnostic accuracy of automated vs manual immature neutrophils for invasive bacterial infections. Although manual counts were superior compared with automated counts, bands had suboptimal accuracy overall and had significant variability in test characteristics based on methodology.

Flow cytometric characterization of hemocytes of the flat oyster, Ostrea chilensis.

The flat oyster, Ostrea chilensis, native to New Zealand (NZ) and Chile is considered an important ecological, cultural and fisheries resource. Currently, commercial landings of this species in NZ are restricted due to low population numbers caused by ongoing mortalities resulting from the presence of the haplosporidian parasite, Bonamia exitiosa. More recently, the arrival of B. ostreae in NZ led to major mortalities in farmed stocks. To understand how diseases caused by Bonamia spp. affect this oyster species, a more complete understanding of its biology, physiology and immune system is needed. The present study characterized, for the first time, hemocytes of adult O. chilensis, from the Foveaux Strait, NZ, using flow cytometry (FCM) and histology. Based on the internal complexity of the hemocytes, two main circulating hemocyte populations were identified: granulocytes and hyalinocytes (accounting for ~30% and ~70% of the total circulating hemocyte population, respectively). These were further divided into two sub-populations of each cell type using FCM. A third sub-population of granulocytes was identified using histology. Using FCM, functional and metabolic characteristics were investigated for the two main hemocyte types. Granulocytes showed higher phagocytic capabilities, lysosomal content, neutral lipid content and reactive oxygen species production compared to hyalinocytes, indicating their important role in cellular immune defence in this species. Methods of hemocyte sampling and storage were also investigated and flow cytometric protocols were detailed and verified to allow effective future investigations into the health status of this important species.

Impairment of G-CSF receptor on granulocytic progenitor cells causes neutropenia in protein malnutrition.

OBJECTIVE: It is well known that protein malnutrition (PM) states can affect hematopoiesis, leading to severe leukopenia and reduced number of granulocytes, which act as the first line of defense, and are important to the innate immune response. The aim of this study was to elucidate some of the mechanisms involved in the impairment of granulopoiesis in PM. METHODS: Male C57BL/6 mice were submitted to PM with a low-protein diet containing 2% protein. Control mice were fed a 12% protein-containing diet. Bone marrow histology and the percentage of granulocytic progenitors were evaluated after in vivo granulocyte-colony stimulating factor (G-CSF) stimulus. Cell proliferation, STAT3 signaling, and the expression of G-CSF receptor were evaluated in hematopoietic progenitor cells. RESULTS: Malnourished animals presented with leukopenia associated with reduced number of granulocytes and reduced percentage of granulocytic progenitors; however, no differences were observed in the regulatory granulopoietic cytokine G-CSF. Additionally, the malnourished group presented with impaired response to in vivo G-CSF stimulus compared with control animals. PM was implicated in decreased ability of c-Kit+ cells to differentiate into myeloid progenitor cells and downregulated STAT3 signaling. Furthermore, the malnourished group exhibited reduced expression of G-CSF receptor on granule-monocytic progenitors. This reduced expression was not completely reversible with G-CSF treatment. CONCLUSIONS: This study implies that PM promotes intrinsic alterations to hematopoietic precursors, which result in hematologic changes, mainly neutropenia, observed in peripheral blood in PM states.

Growth hormone on ovarian morphology of lambaris (Astyanax bimaculatus) after induced spawning / Hormônio de crescimento sobre a morfologia ovariana de lambaris (Astyanax bimaculatus) após desova induzida

Lambari, Astyanax bimaculatus, is an oviparous, multiple-spawning fish that is reproductively active throughout the year, which makes it promising for cultivation and research. This research histologically evaluates the ovaries of lambari that have undergone artificial spawning induced with pituitary extract (control group), and the effect of growth hormone at a dose of 2 mg/g body weight (treatment group) on the subsequent process of ovarian recovery. Ovaries of fish in both the control and treatments groups were collected at 120 hours after spawning and analyzed using optical microscopy to characterize the average quantities of: follicles in different stages of development, post-ovulatory follicles, follicular atresia and granulocytes. Quantity and morphology of early and advanced primary follicles did not differ between the treatment and control groups; an important and necessary factor for ovarian recovery for subsequent spawning. There was a greater amount of granulocytes in initial atresia in the group treated with growth hormone. These results demonstrated that the administration of growth hormone may potentiate the process of ovarian recovery after induced spawning.(AU)

O Lambari Astyanax bimaculatus é um peixe ovíparo de desova múltipla que é reprodutivamente ativo durante todo o ano, o que o torna promissor para cultivo e pesquisa. Este trabalho avalia histologicamente os ovários de lambaris submetidos à desova artificial, induzida pelo extrato hipofisário (grupo controle) e o efeito do hormônio de crescimento na concentração de 2 g/g de massa corporal (grupo tratamento) no subsequente processo de recuperação ovariana. Os ovários dos peixes dos grupos controle e tratamento foram coletados às 120 horas após a desova e analisados em microscopia óptica para caracterizar as quantidades médias de: folículos em diferentes estágios de desenvolvimento, folículos pós-ovulatórios, atresia folicular e granulócitos. A quantidade e a morfologia dos folículos primários iniciais e avançados não diferiram entre os grupos tratamento e controle; um fator importante e necessário para a recuperação dos ovários para posterior desova. Houve maior quantidade de granulócitos na atresia inicial no grupo tratado com hormônio de crescimento. Esses resultados demonstram que a administração do hormônio do crescimento pode potencializar o processo de recuperação ovariana após a desova induzida.(AU)

Ex vivo Manufactured Neutrophils for Treatment of Neutropenia-A Process Economic Evaluation.

Neutropenia is a common side-effect of acute myeloid leukemia (AML) chemotherapy characterized by a critical drop in neutrophil blood concentration. Neutropenic patients are prone to infections, experience poorer clinical outcomes, and require expensive medical care. Although transfusions of donor neutrophils are a logical solution to neutropenia, this approach has not gained clinical traction, primarily due to challenges associated with obtaining sufficiently large numbers of neutrophils from donors whilst logistically managing their extremely short shelf-life. A protocol has been developed that produces clinical-scale quantities of neutrophils from hematopoietic stem and progenitor cells (HSPC) in 10 L single-use bioreactors (1). This strategy could be used to mass produce neutrophils and generate sufficient cell numbers to allow decisive clinical trials of neutrophil transfusion. We present a bioprocess model for neutrophil production at relevant clinical-scale. We evaluated two production scenarios, and the impact on cost of goods (COG) of multiple model parameters including cell yield, materials costs, and process duration. The most significant contributors to cost were consumables and raw materials, including the cost of procuring HSPC-containing umbilical cord blood. The model indicates that the most cost-efficient culture volume (batch size) is ~100 L in a single bioreactor. This study serves as a framework for decision-making and optimization strategies when contemplating the production of clinical quantities of cells for allogeneic therapy.

Hematological Parameters as Prognostic Biomarkers in Patients with Cardiovascular Diseases.

Cardiovascular diseases are the major causes of preventable health loss from disease in the world and lead to functional disturbances including hematological parameters. The inflammatory and hypoxemic nature of cardiovascular diseases causes a stimulus in the bone marrow and, depending on the intensity of this stimulus, there is a release of immature cells or increase of other cells in the bloodstream. Therefore, their presence in the circulation is an important variable used to diagnose, stratify and predict diseases. In the last five decades, with the advent of automated counting of immature cells in the peripheral blood, the hemogram was transformed into a clinical tool of great importance in hospital surveillance for demonstrating this daily variability in the hematopoietic response according to the existing injury in the patient. Studies have shown that the presence of nucleated red blood cells and increases in mean platelet volume, immature granulocytes and neutrophil to lymphocyte ratio in the systemic circulation are independent prognostic biomarkers. This review article has as main objective to demonstrate the association of these hematological parameters to cardiovascular diseases, emphasizing their importance in clinical decision making.

Leptospira species promote a pro-inflammatory phenotype in human neutrophils.

Leptospirosis is a global zoonosis caused by pathogenic Leptospira. Neutrophils are key cells against bacterial pathogens but can also contribute to tissue damage. Because the information regarding the role of human neutrophils in leptospirosis is scant, we comparatively analysed the human neutrophil's response to saprophytic Leptospira biflexa serovar Patoc (Patoc) and the pathogenic Leptospira interrogans serovar Copenhageni (LIC). Both species triggered neutrophil responses involved in migration, including the upregulation of CD11b expression, adhesion to collagen, and the release of IL-8. In addition, both species increased levels of pro-inflammatory IL-1ß and IL-6 associated with the inflammasome and NFκB pathway activation and delayed neutrophil apoptosis. LIC was observed on the neutrophil surface and not phagocytized. In contrast, Patoc generated intracellular ROS associated with its uptake. Neutrophils express the TYRO3, AXL, and MER receptor protein tyrosine kinases (TAM), but only LIC selectively increased the level of AXL. TLR2 but not TLR4-blocking antibodies abrogated the IL-8 secretion triggered by both Leptospira species. In summary, we demonstrate that Leptospira species trigger a robust neutrophil activation and pro-inflammatory response. These findings may be useful to find new diagnostic markers and therapeutic strategies against leptospirosis.

Growth hormone on ovarian morphology of lambaris (Astyanax bimaculatus) after induced spawning / Hormônio de crescimento sobre a morfologia ovariana de lambaris (Astyanax bimaculatus) após desova induzida

ABSTRACT: Lambari, Astyanax bimaculatus, is an oviparous, multiple-spawning fish that is reproductively active throughout the year, which makes it promising for cultivation and research. This research histologically evaluates the ovaries of lambari that have undergone artificial spawning induced with pituitary extract (control group), and the effect of growth hormone at a dose of 2 mg/g body weight (treatment group) on the subsequent process of ovarian recovery. Ovaries of fish in both the control and treatments groups were collected at 120 hours after spawning and analyzed using optical microscopy to characterize the average quantities of: follicles in different stages of development, post-ovulatory follicles, follicular atresia and granulocytes. Quantity and morphology of early and advanced primary follicles did not differ between the treatment and control groups; an important and necessary factor for ovarian recovery for subsequent spawning. There was a greater amount of granulocytes in initial atresia in the group treated with growth hormone. These results demonstrated that the administration of growth hormone may potentiate the process of ovarian recovery after induced spawning.

RESUMO: O Lambari Astyanax bimaculatus é um peixe ovíparo de desova múltipla que é reprodutivamente ativo durante todo o ano, o que o torna promissor para cultivo e pesquisa. Este trabalho avalia histologicamente os ovários de lambaris submetidos à desova artificial, induzida pelo extrato hipofisário (grupo controle) e o efeito do hormônio de crescimento na concentração de 2 μg/g de massa corporal (grupo tratamento) no subsequente processo de recuperação ovariana. Os ovários dos peixes dos grupos controle e tratamento foram coletados às 120 horas após a desova e analisados em microscopia óptica para caracterizar as quantidades médias de: folículos em diferentes estágios de desenvolvimento, folículos pós-ovulatórios, atresia folicular e granulócitos. A quantidade e a morfologia dos folículos primários iniciais e avançados não diferiram entre os grupos tratamento e controle; um fator importante e necessário para a recuperação dos ovários para posterior desova. Houve maior quantidade de granulócitos na atresia inicial no grupo tratado com hormônio de crescimento. Esses resultados demonstram que a administração do hormônio do crescimento pode potencializar o processo de recuperação ovariana após a desova induzida.

Neutrophils, eosinophils, and mast cells in the intestinal wall of dogs naturally infected with Leishmania infantum / Neutrófilos, eosinófilos e mastócitos na parede intestinal de cães naturalmente infectados com Leishmania infantum

Abstract Visceral leishmaniasis (VL) is a disease caused by the protozoa Leishmania infantum and can cause an inflammatory reaction in the gastrointestinal tract, however the role of granulocytic cells (neutrophils, eosinophils, and mast cells) in the intestine of dogs infected is not fully understood. We performed a quantitative analysis these cells in the intestinal wall of dogs with canine visceral leishmaniasis (CVL). Twenty dogs were assigned to one of three groups: group 1 (G1, n=8), dogs with CVL and L. infantum amastigotes in the intestine; group 2 (G2, n=9), dogs with CVL but without intestinal amastigotes; and group 3 (G3, n=3), uninfected dogs (control group). Granulocytic cells were counted in the crypt-villus unit (mucosa), submucosa, and muscle layer of the intestinal mucosa. Cell counts were higher in the intestinal wall of dogs from G2 followed by G1 and G3 (p≤0.05). In G1, there was a low inverse correlation between parasite burden of the small intestine and granulocyte counts (r= -0.1, p≤0.01). However, in G2 dogs, mast cell and eosinophil numbers showed positive correlation (r=0.85, p≤0.01). The granulocytic cell hyperplasia observed in the intestine of L. infantum-infected dogs suggests that these cells may be involved in the cell-mediated immune response for parasite elimination.

Resumo A leishmaniose visceral (LV) é uma doença causada pelo protozoário Leishmania infantum e pode causar uma reação inflamatória no trato gastrointestinal, entretanto o papel das células granulocíticas (neutrófilos, eosinófilos e mastócitos) no intestino de cães infectados não é totalmente compreendido. Neste estudo realizamos uma análise quantitativa dessas células na parede intestinal de cães com LV. Vinte cães foram distribuídos em três grupos: grupo 1 (G1, n=8), cães com LV e amastigotas de L. infantum no intestino; grupo 2 (G2, n=9), cães com LV, mas sem amastigotas intestinais; e grupo 3 (G3, n=3), não infectados (grupo controle). Células granulocíticas foram contadas na unidade cripta-vilo (mucosa), submucosa e camada muscular da mucosa intestinal. Observamos hiperplasia dessas células na parede intestinal de cães do G2, seguidas das G1 em relação ao G3 (p≤0,05). No G1, houve uma correlação inversa baixa entre a carga parasitária do intestino delgado e a contagem de granulócitos (r= -0,1; p≤0,01). No entanto, nos cães do G2, os números de mastócitos e eosinófilos apresentaram correlação positiva (r=0,85; p≤0,01). A hiperplasia de células granulocíticas observada no intestino de cães infectados por L. infantum sugere que essas células podem estar envolvidas na resposta imune mediada por células para a eliminação do parasita.

Neutrophils, eosinophils, and mast cells in the intestinal wall of dogs naturally infected with Leishmania infantum / Neutrófilos, eosinófilos e mastócitos na parede intestinal de cães naturalmente infectados com Leishmania infantum

Visceral leishmaniasis (VL) is a disease caused by the protozoa Leishmania infantum and can cause an inflammatory reaction in the gastrointestinal tract, however the role of granulocytic cells (neutrophils, eosinophils, and mast cells) in the intestine of dogs infected is not fully understood. We performed a quantitative analysis these cells in the intestinal wall of dogs with canine visceral leishmaniasis (CVL). Twenty dogs were assigned to one of three groups: group 1 (G1, n=8), dogs with CVL and L. infantum amastigotes in the intestine; group 2 (G2, n=9), dogs with CVL but without intestinal amastigotes; and group 3 (G3, n=3), uninfected dogs (control group). Granulocytic cells were counted in the crypt-villus unit (mucosa), submucosa, and muscle layer of the intestinal mucosa. Cell counts were higher in the intestinal wall of dogs from G2 followed by G1 and G3 (p≤0.05). In G1, there was a low inverse correlation between parasite burden of the small intestine and granulocyte counts (r= -0.1, p≤0.01). However, in G2 dogs, mast cell and eosinophil numbers showed positive correlation (r=0.85, p0.01). The granulocytic cell hyperplasia observed in the intestine of L. infantum-infected dogs suggests that these cells may be involved in the cell-mediated immune response for parasite elimination.(AU)

A leishmaniose visceral (LV) é uma doença causada pelo protozoário Leishmania infantum e pode causar uma reação inflamatória no trato gastrointestinal, entretanto o papel das células granulocíticas (neutrófilos, eosinófilos e mastócitos) no intestino de cães infectados não é totalmente compreendido. Neste estudo realizamos uma análise quantitativa dessas células na parede intestinal de cães com LV. Vinte cães foram distribuídos em três grupos: grupo 1 (G1, n=8), cães com LV e amastigotas de L. infantum no intestino; grupo 2 (G2, n=9), cães com LV, mas sem amastigotas intestinais; e grupo 3 (G3, n=3), não infectados (grupo controle). Células granulocíticas foram contadas na unidade cripta-vilo (mucosa), submucosa e camada muscular da mucosa intestinal. Observamos hiperplasia dessas células na parede intestinal de cães do G2, seguidas das G1 em relação ao G3 (p0,05). No G1, houve uma correlação inversa baixa entre a carga parasitária do intestino delgado e a contagem de granulócitos (r= -0,1; p≤0,01). No entanto, nos cães do G2, os números de mastócitos e eosinófilos apresentaram correlação positiva (r=0,85; p≤0,01). A hiperplasia de células granulocíticas observada no intestino de cães infectados por L. infantum sugere que essas células podem estar envolvidas na resposta imune mediada por células para a eliminação do parasita.(AU)