Bruno Cogliati

University of São Paulo, San Paulo, São Paulo, Brazil

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Publications (48)99.77 Total impact

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    ABSTRACT: Cellular channels composed of connexin 43 are known to act as key players in the life cycle of the skin and consequently to underlie skin repair. This study was specifically set up to investigate the suite of molecular mechanisms driven by connexin 43-based channels on wound healing. To this end, a battery of parameters, including re-epithelialization, neovascularization, collagen deposition and extracellular matrix remodeling, was monitored over time during experimentally induced skin repair in heterozygous connexin 43 knockout mice. It was found that connexin 43 deficiency accelerates re-epithelialization and wound closure, increases proliferation and activation of dermal fibroblasts, and enhances the expression of extracellular matrix remodeling mediators. These data substantiate the notion that connexin 43 may represent an interesting therapeutic target in dermal wound healing. Copyright © 2015 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.
    Journal of dermatological science 04/2015; DOI:10.1016/j.jdermsci.2015.03.019 · 3.34 Impact Factor
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    ABSTRACT: Liver fibrosis occurring as an outcome of non-alcoholic steatohepatitis (NASH) can precede the development of cirrhosis. We investigated the effects of sorafenib in preventing liver fibrosis in a rodent model of NASH. Adult Sprague-Dawley rats were fed a choline-deficient high-fat diet and exposed to diethylnitrosamine for 6 weeks. The NASH group (n=10) received vehicle and the sorafenib group (n=10) received 2.5 mg·kg-1·day-1 by gavage. A control group (n=4) received only standard diet and vehicle. Following treatment, animals were sacrificed and liver tissue was collected for histologic examination, mRNA isolation, and analysis of mitochondrial function. Genes related to fibrosis (MMP9, TIMP1, TIMP2), oxidative stress (HSP60, HSP90, GST), and mitochondrial biogenesis (PGC1α) were evaluated by real-time quantitative polymerase chain reaction (RT-qPCR). Liver mitochondrial oxidation activity was measured by a polarographic method, and cytokines by enzyme-linked immunosorbent assay (ELISA). Sorafenib treatment restored mitochondrial function and reduced collagen deposition by nearly 63% compared to the NASH group. Sorafenib upregulated PGC1α and MMP9 and reduced TIMP1 and TIMP2 mRNA and IL-6 and IL-10 protein expression. There were no differences in HSP60, HSP90 and GST expression. Sorafenib modulated PGC1α expression, improved mitochondrial respiration and prevented collagen deposition. It may, therefore, be useful in the treatment of liver fibrosis in NASH.
    Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas / Sociedade Brasileira de Biofisica ... [et al.] 02/2015; DOI:10.1590/1414-431X20143962 · 1.08 Impact Factor
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    ABSTRACT: Hepatotoxicity, including drug-induced liver injury, is frequently accompanied by cell death. The latter is typically driven by apoptosis or necrosis, which substantially differ based upon biochemical and morpho- logical criteria. This chapter describes two commonly used methods to probe apoptotic and necrotic activi- ties in adherent monolayer cultures of primary hepatocytes. The apoptosis assay uses a prototypical substrate of caspase 3, the main executor of apoptotic cell death, which can be cleaved, yielding a product that can be measured fl uorimetrically. The second assay relies on the disruption of the cell plasma mem- brane, which typically occurs in necrotic cell death and that results in the extracellular release of cytoplas- mic enzymes, such as lactate dehydrogenase. The latter can be indirectly assessed by spectrophotometrically measuring the consumption of reduced nicotinamide adenine dinucleotide.
    Protocols in In Vitro Hepatocyte Research, 2015 edited by Mathieu Vinken, Vera Rogiers, 01/2015: chapter Measurement of Apoptotic and Necrotic Cell Death in Primary Hepatocyte Cultures: pages 349-362; Springer Protocols-Humana Press., ISBN: 978-1-4939-2073-0
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    ABSTRACT: Connexins are proteins that form gap junctions. Perturbations in the cell membrane reportedly promote changes in the expression profile of connexins. Electroporation promotes destabilization by applying electrical pulses, and this procedure is used in electrochemotherapy and gene therapy, among others. This in vitro work aimed to study the interference of electroporation on the expression profile of GJB2 (Cx26 gene) and Connexin 26 in melanoma cell line B16/BL6. The techniques of immunocytochemistry, Western blot, and real-time PCR were used. After electroporation, cells showed a transient decrease in GJB2 mRNA. The immunostaining of Cx26 showed no noticeable change after electroporation at different time points. However, Western blot showed a significant reduction in Cx26 30 min after electroporation. Our results showed that electroporation interferes transiently in the expression of Connexin 26 in melanoma and are consistent with the idea that electroporation is a process of intense stress that promotes cell homeostatic imbalance and results in disruption of cell physiological processes such as transcription and translation.
    Journal of Membrane Biology 10/2014; 248(1). DOI:10.1007/s00232-014-9735-z · 2.17 Impact Factor
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    ABSTRACT: Background: Very few studies have evaluated the expression of homeobox A10 (HOXA10) and steroid (estrogen and progesterone) receptors exclusively in deep endometriosis. Conclusions drawn from studies evaluating peritoneal and ovarian endometriosis are usually generalized to explain the pathogenesis of the disease as a whole. We aimed to evaluate the expression of HOXA10, estrogen receptor (ER-), progesterone receptor (PR), and PR-B in rectosigmoid endometriosis (RE), a typical model of deep disease. Methods: We used RE samples from 18 consecutive patients to construct tissue microarray blocks. Nine patients each were operated during the proliferative and secretory phases of the menstrual cycle. We quantified the expressions of proteins by immunohistochemistry using the modified Allred score. Result: The HOXA10 was expressed in the stroma of nodules during the secretory phase in 5 of the 18 patients. Expression of ER- (in 16 of 18 patients), PR (in 17 of 18 patients), and PR-B (17 of 18 patients) was moderate to strong in the glands and stroma of nodules during both phases. Expression of both PR (P = .023) and PR-B (P = .024) was significantly greater during the secretory phase. Conclusion: The HOXA10 is expressed in RE, where it likely imparts the de novo identity of endometriotic lesions. The ER-, PR, and PR-B are strongly expressed in RE, which differs from previous studies investigating peritoneal and ovarian lesions. This suggests different routes of pathogenesis for each of the 3 types of endometriosis.
    Reproductive sciences (Thousand Oaks, Calif.) 09/2014; 22(1). DOI:10.1177/1933719114549846 · 2.18 Impact Factor
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    ABSTRACT: Background: Melanoma is one of the most common skin neoplasms in humans and dogs. The tumor microenvironment in melanoma comprises cancer cells and stromal cells that interact to accelerate tumor progression. Several prognostic markers for melanomas have been studied in many human tumors, including fibroblast-specific protein 1 (S100A4). S100A4 is a member of the S100 family of calcium-binding proteins in stromal cells.Hypothesis/objectives: The objective of this study was to describe the immunohistochemical patterns of S100A4 in stroma and neoplastic cells of canine skin melanomas and correlate them with some histological parameters. Animals and Methods: Forty-eight samples (38 pigmented and 10 non-pigmented melanomas) were first selected and their nature confirmed using S100, Melan A and vimentin. All cases were examined by immunohistochemistry using S100A4 to correlate expression, histotype, and level of invasion.Results: All the tumors, including 10 non-pigmented, were positive for S100, Melan A, vimentin and negative for cytokeratin AE1/AE3 (consistent with melanomas). The 48 melanomas were classified as epithelioid (n = 21), spindle (n = 14), and mixed (n = 13). S100A4 was preferentially expressed in epithelioid and spindle cell types compared with mixed melanomas and S100A4 expression was not associated with level of invasion (Clark's levels IV to V).Conclusion: S100A4 expression in melanoma samples varied among histotypes but not between levels of invasion.
    The Veterinary quarterly 07/2014; 34(2):1-7. DOI:10.1080/01652176.2014.936628 · 0.65 Impact Factor
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    ABSTRACT: Lung cancer is the leading cause of cancer-related mortality in both men and women throughout the world. This disease is strongly associated with tobacco smoking. The aim of this manuscript was to establish an in vitro model that mimics the chronic exposures of alveolar epithelial type II cells to the tobacco-specific nitrosamine carcinogen, NNK. Immortalized non-neoplastic alveolar epithelial cells type II, (E10 cells), from BALB/c mice were exposed to low concentration of NNK (100 pM) during 5, 10, 15, and 20 cycles of 48 h. NNK-transformed cells showed an increase of proliferation rate and motility. Moreover, these cells underwent epithelial-to-mesenchymal transition (EMT). Increased migratory capacity and EMT were correlated to the time of exposure to NNK. NNK-transformed cells were tested for their growth and metastatic capacity in vivo. Subcutaneous injection of cells exposed to NNK for 20 cycles (E10-NNK20 clone) into BALB/c mice led to the formation of subcutaneous tumors that arose after 40 ± 17 d in all animals, which died 95 ± 18 d after cell inoculation, with lymph nodes and lung metastasis. The morphological characteristics of tumors were compatible with metastatic undifferentiated carcinoma. Cells exposed to NNK for 5-10 cycles did not display metastatic capacity, while those exposed for 15 cycles displayed low capacity. Our results show that prolonged exposures to NNK led the cells to increasingly acquire malignant properties. The cellular model presented in this study is suitable for studying the molecular events involved in the different stages of malignant transformation. © 2012 Wiley Periodicals, Inc.
    Molecular Carcinogenesis 05/2014; 53(5). DOI:10.1002/mc.21987 · 4.77 Impact Factor
  • Journal of Hepatology 04/2014; 60(1):S152. DOI:10.1016/S0168-8278(14)60424-4 · 10.40 Impact Factor
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    ABSTRACT: Rationale: The excessive intake of vitamin A in the form of vitamin concentrate, supplement or vitamin-rich liver can result in hypervitaminosis A in man and animals. Although osteopathologies resulting from chronic vitamin A intoxication in cats are well characterized, no information is available concerning feline hypervitaminosis A-induced liver disease. Clinical summary: We report the first case of hepatic stellate cell lipidosis and hepatic fibrosis in a domestic cat that had been fed a diet based on raw beef liver. Radiographic examination revealed exostoses and ankylosis between vertebrae C1 and T7, compatible with deforming cervical spondylosis. Necropsy showed a slightly enlarged and light yellow to bronze liver. Microscopic and ultrastructural analyses of liver tissues revealed diffuse and severe liver fibrosis associated with hepatic stellate cell hyperplasia and hypertrophy. These cells showed immunopositive staining for α-smooth muscle actin and desmin markers. The necropsy findings of chronic liver disease coupled with osteopathology supported the diagnosis of hypervitaminosis A. Practical relevance: As in human hepatology, if there is dietary evidence to support increased intake of vitamin A, then hypervitaminosis A should be considered in the differential diagnosis of chronic liver disease in cats.
    03/2014; 16(3):243-8. DOI:10.1177/1098612X13516121
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    ABSTRACT: Hair follicle tumours generally present as benign, solitary masses and have a good prognosis following surgical resection. This report describes a case of multiple trichoblastomas in a dog. A 2-year-old crossbred dog presented with multiple soft cutaneous periocular, perilabial, submandibular and nasal nodules, between 2 and 9 cm in diameter, located on the right side of the face. New nodules were observed on the same side of the face at a second consultation 3 weeks later. Surgical resection of all nodules was performed in two procedures. Three nodules were initially resected and submitted for histolopathology and immunohistochemistry. The diagnosis was trichoblastoma for all three. At the time of the second consultation, new and remaining nodules were biopsied and the diagnosis of trichoblastoma confirmed. The dog was treated with doxorubicin and piroxicam for 30 days prior to the second surgical procedure in an attempt to reduce new tumour growth and the size of present tumours. All nodules were resected and the defects closed using rotation flaps. No recurrence of the neoplasm was noted within 10 months after surgery. Trichoblastomas are generally benign but can present as multiple neoplasms that may require surgical resection and may respond to chemotherapy. To the authors' knowledge, this is the first report of multiple trichoblastomas in a dog.
    Veterinary Dermatology 02/2014; 25(1):48-e19. DOI:10.1111/vde.12100 · 1.99 Impact Factor
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    ABSTRACT: The liver was among the first organs in which connexin proteins have been identified. Hepatocytes harbor connexin32 and connexin26, while non-parenchymal liver cells typically express connexin43. Connexins give rise to hemichannels, which dock with counterparts on adjacent cells to form gap junctions. Both hemichannels and gap junctions provide pathways for communication, via paracrine signaling or direct intercellular coupling, respectively. Over the years, hepatocellular gap junctions have been shown to regulate a number of liver-specific functions and to drive liver cell growth. In the last few years, it has become clear that connexin hemichannels are involved in liver cell death, particularly in hepatocyte apoptosis. This also holds true for hemichannels composed of pannexin1, a connexin-like protein recently identified in the liver. Moreover, pannexin1 hemichannels are key players in the regulation of hepatic inflammatory processes. The current paper provides a concise overview of the features of connexins, pannexins and their channels in the liver.
    Frontiers in Physiology 01/2014; 4:405. DOI:10.3389/fphys.2013.00405
  • 01/2014; 31(1):33-41. DOI:10.4322/jms.ao060313
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    ABSTRACT: Studies of skin wound healing in crocodilians are necessary given the frequent occurrence of cannibalism in intensive farming systems. Air temperature affects tissue recovery because crocodilians are ectothermic. Therefore, the kinetics of skin wound healing in Caiman yacare were examined at temperatures of 33°C and 23°C. Sixteen caiman were selected and divided into two groups of eight maintained at 23°C or 33°C. The studied individuals' scars were photographed after 1, 2, 3, 7, 15 and 30 days of the experimental conditions, and samples were collected for histological processing after 3, 7, 15 and 30 days. Macroscopically, the blood clot (heterophilic granuloma) noticeably remained in place covering the wound longer for the caiman kept at 23°C. Microscopically, the temperature of 23°C slowed epidermal migration and skin repair. Comparatively, new blood vessels, labeled using von Willebrand factor (vWF) antibody staining, were more frequently found in the scars of the 33°C group. The collagen fibers in the dermis were denser in the 33°C treatment. Considering the delayed healing at 23°C, producers are recommended to keep wounded animals at 33°C, especially when tanks are cold, to enable rapid wound closure and better repair of collagen fibers because such lesions tend to compromise the use of their skin as leather.
    11/2013; 2(11):1171-8. DOI:10.1242/bio.20135876
  • Journal of Minimally Invasive Gynecology 11/2013; 20(6):S29-S30. DOI:10.1016/j.jmig.2013.08.093 · 1.58 Impact Factor
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    ABSTRACT: Melanoma is a malignant neoplasm occurring in several animal species, and is the most frequently found tumor in the oral cavity in dogs. Melanomas are classified into two types: melanotic and amelanotic. Prior research suggests that human amelanotic melanomas are more aggressive than their melanotic counterparts. This study evaluates the behavior of canine melanotic and amelanotic oral cavity melanomas and quantifies cell proliferation and the expression of connexins. Twenty-five melanomas (16 melanotic and 9 amelanotic) were collected from dogs during clinical procedures at the Veterinary Hospital of the School of Veterinary Medicine and Animal Science of the University of São Paulo, Brazil. After diagnosis, dogs were followed until death or euthanasia. Histopathology confirmed the gross melanotic or amelanotic characteristics and tumors were classified according to the WHO. HMB45 or Melan A immunostainings were performed to confirm the diagnosis of amelanotic melanomas. Cell proliferation was quantified both by counting mitotic figures and PCNA positive nuclei. Expressions of connexins 26 and 43 were evaluated by immunohistochemistry, qRT-PCR and Western blot. Dogs bearing amelanotic melanomas presented a shorter lifespan in comparison to those with melanotic melanomas. Cell proliferation was significantly higher in amelanotic melanomas. Expressions of Connexins 26 and 43 were significantly reduced in amelanotic melanomas. The results presented here suggest that oral cavity melanotic and amelanotic melanomas differ regarding their behavior, cell proliferation and connexin expression in dogs, indicating a higher aggressiveness of amelanotic variants.
    Veterinary Research Communications 10/2013; 38(1). DOI:10.1007/s11259-013-9580-z · 1.36 Impact Factor
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    ABSTRACT: Gap junctions are communicating junctions which are important for tissue homeostasis, and their disruption is involved in carcinogenic processes. This study aimed to verify the influence of deletion of one allele of the Connexin 43 gene on cancer incidence in different organs. The 7, 12-dimethylbenzanthracene (DMBA) carcinogenic model, using hebdomadary doses by gavage of 9 mg per animal, was used to induce tumors in Connexin 43 heterozygous or wild-type mice. The experiment began in the eighth week of the mice life, and all of them were euthanized when reaching inadequate physical condition, or at the end of 53 weeks. No statistical differences occurred for weight gain and cancer survival time (P = 0.9853) between heterozygous and wild-type mice. Cx43(+/-) mice presented significantly higher susceptibility to lung cancer (P = 0.0200) which was not evidenced for benign neoplasms (P = 0.3449). In addition, incidence of ovarian neoplasms was 2.5-fold higher in Cx43(+/-) mice, although not statistically significant. Other organs showed a very similar cancer occurrence between Cx43 groups. The experiment strengthens the evidence of the relationship between Connexin 43 deficiency and carcinogenesis.
    10/2013; 2013:618475. DOI:10.1155/2013/618475
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    ABSTRACT: Apoptosis not only plays a key role in physiological demise of defunct hepatocytes, but is also associated with a plethora of acute and chronic liver diseases as well as with hepatotoxicity. The present paper focuses on the modelling of this mode of programmed cell death in primary hepatocyte cultures. Particular attention is paid to the activation of spontaneous apoptosis during the isolation of hepatocytes from the liver, its progressive manifestation upon the subsequent establishment of cell cultures and simultaneously to strategies to counteract this deleterious process. In addition, currently applied approaches to experimentally induce controlled apoptosis in this in vitro setting for mechanistic research purposes and thereby its detection using relevant biomarkers are reviewed.
    Archives of Toxicology 09/2013; 88(2). DOI:10.1007/s00204-013-1123-4 · 5.08 Impact Factor
  • Journal of Feline Medicine & Surgery 09/2013; 15(9):818-28. DOI:10.1177/1098612X13500432 · 1.22 Impact Factor
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    ABSTRACT: S-Nitroso-N-acetylcysteine (SNAC) is a water soluble primary S-nitrosothiol capable of transferring and releasing nitric oxide and inducing several biochemical activities, including modulation of hepatic stellate cell activation. In this study, we evaluated the antifibrotic activity of SNAC in an animal model of nonalcoholic steatohepatitis (NASH) induced in Sprague-Dawley rats fed with a choline-deficient, high trans fat diet and exposed to diethylnitrosamine for 8 weeks. The rats were divided into three groups: SNAC, which received oral SNAC solution daily; NASH, which received the vehicle; and control, which received standard diet and vehicle. Genes related to fibrosis (matrix metalloproteinases [MMP]-13, -9, and -2), transforming growth factor β-1 [TGFβ-1], collagen-1α, and tissue inhibitors of metalloproteinase [TIMP-1 and -2] and oxidative stress (heat-shock proteins [HSP]-60 and -90) were evaluated. SNAC led to a 34.4% reduction in the collagen occupied area associated with upregulation of MMP-13 and -9 and downregulation of HSP-60, TIMP-2, TGFβ-1, and collagen-1α. These results indicate that oral SNAC administration may represent a potential antifibrotic treatment for NASH.
    Drug Design, Development and Therapy 06/2013; 7:553-563. DOI:10.2147/DDDT.S43930 · 3.03 Impact Factor
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    ABSTRACT: Background/objective The early detection of focal hepatic lesions using ultrasound scanning is challenging, and this challenge becomes even greater in the presence of diffuse parenchymal disease. This study aimed to evaluate the diagnostic performance of elastography and contrast-enhanced ultrasonography (CEUS) in the early detection of hepatocellular lesions in an experimental rat model of nonalcoholic steatohepatitis (NASH). Methods B-mode and Doppler ultrasonography was performed weekly in 30 rats divided into a NASH group (n = 20) and a group without liver disease (n = 10). The animals underwent elastography and CEUS and were then euthanized. Liver nodules were assessed by histopathology. Results Doppler mapping results of lesions with vascularization were considered indicative of malignancy, with a sensitivity of 29% before and 71% after contrast injection. The specificity was 71% before and 96% after CEUS. Elastograms of positive lesions showed areas of high stiffness, which were indicative of malignancy. This malignancy was confirmed by the histologic evaluation, with a sensitivity of 90% and a specificity of 60%. After CEUS analysis, 4 nodules were identified that were not observed on B-mode ultrasonography. Early wash-in was significantly associated with malignancy (sensitivity of 88% and specificity of 67%). Conclusions Both techniques allow for the correct diagnosis of well-differentiated to moderately differentiated hepatocellular carcinomas with good accuracy in an experimental rat model of NASH.
    06/2013; 3(2):96–101. DOI:10.1016/j.jceh.2013.04.004

Publication Stats

83 Citations
99.77 Total Impact Points

Institutions

  • 2008–2015
    • University of São Paulo
      • • Departamento de Psicologia
      • • Department of Veterinary Medicine (ZMV)
      • • Departamento de Patologia (FM) (São Paulo)
      San Paulo, São Paulo, Brazil
  • 2010
    • Senac São Paulo
      San Paulo, São Paulo, Brazil