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Most of the obesity-related complications are due to ectopic fat accumulation. Recently, the activation of the cell-surface receptor for advanced glycation end products (RAGE) has been associated with lipid accumulation in different organs. Nevertheless, the role of RAGE and sRAGE, the soluble form that prevents ligands to activate RAGE, in intramyocardial lipid accumulation is presently unknown. To this aim, we analyzed whether, in obesity, intramyocardial lipid accumulation and lipid metabolism-related transcriptome are related to RAGE and sRAGE. Heart and serum samples were collected from 10 lean (L) and 10 obese (OB) Zucker rats. Oil red staining was used to detect lipids on frozen heart sections. The lipid metabolism-related transcriptome (84 genes) was analyzed by a specific PCR array. Heart RAGE expression was explored by real-time RT-PCR and Western blot analyses. Serum levels of sRAGE (total and endogenous secretory form (esRAGE)) were quantified by ELISA. Genes promoting fatty acid transport, activation, and oxidation in mitochondria/peroxisomes were upregulated in OB hearts. Intramyocardial lipid content did not differ between OB and L rats, as well as RAGE expression. A slight increase in epicardial adipose tissue was observed in OB hearts. Total sRAGE and esRAGE concentrations were significantly higher in OB rats. sRAGE may protect against obesity-induced intramyocardial lipid accumulation by preventing RAGE hyperexpression, therefore allowing lipids to be metabolized. EAT also played a protective role by working as a buffering system that protects the myocardium against exposure to excessively high levels of fatty acids. These observations reinforce the potential role of RAGE pathway as an interesting therapeutic target for obesity-related complications, at least at the cardiovascular level.
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Produtos Finais de Glicação Avançada/metabolismo , Metabolismo dos Lipídeos/fisiologia , Miocárdio/metabolismo , Obesidade/metabolismo , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Animais , Biomarcadores , Western Blotting , Ensaio de Imunoadsorção Enzimática , Lipídeos , Masculino , Obesidade/sangue , Ratos , Ratos Zucker , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Methicillin-resistant Staphylococcus aureus (MRSA) clones are rapidly increasing beyond the hospital into the community, livestock farming and environmental settings. An Italian man, a professional diver working in Egypt, was admitted to Infectious Diseases Clinic-ASST Fatebenefratelli Sacco for ulcerative skin lesions. An MRSA strain was isolated from the lesions' purulent exudate and the nasal colonization was also ascertained. The strain, characterized by whole genome sequencing, resulted to be Panton-Valentine Leukocidin (PVL) positive, SCCmecI - spa-type t504, and belonging to the sequence type 1153, sporadically described worldwide.
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Genoma Bacteriano , Staphylococcus aureus Resistente à Meticilina , Infecções Comunitárias Adquiridas/microbiologia , Genoma Bacteriano/genética , Genômica , Humanos , Itália , Leucocidinas/metabolismo , Masculino , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Pessoa de Meia-Idade , Infecções Estafilocócicas/microbiologiaRESUMO
The gut-liver axis represents a current topic in human medicine. Extensive research investigates the gut microbiome (GM) modifications in relation to various kinds of chronic hepatobiliary diseases (CHD), with many mechanisms and therapeutical implications recognized. Those aspects in veterinary medicine are still quite unexplored. The aim of the present study was to evaluate GM in dogs diagnosed with CD. Comparison among CHD dogs were made considering some clinical and biochemical variables (lipemia and alanine-aminotransferase activities), presence of cholestasis or endocrine disorders, diet). Sixty-five dogs were prospectively enrolled with clinical and hematobiochemical evaluation and 16S-RNA GM sequencing assessed. Dogs that received antibiotics and/or pre/pro/symbiotics administration were excluded. Deeper GM alteration was observed between dogs with or without ultrasonographic and biochemical cholestatic CHD. Cholestasis was associated with a decrease in several bacterial taxa, including Clostridium hiranonis, Fusobacterium, Megamonas, Ruminococcus faecis, Turicibacter, and higher levels of Escherichia/Shigella and Serratia. Thus, the alteration in bile flow and composition, typical of cholestasis, may directly affect the local intestinal microbial environment. For the management of dogs with CHD and especially cholestatic CHD, clinicians should be aware that gut-liver interaction may lead to dysbiosis.
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An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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PURPOSE: Assessment of inflammatory bowel disease (IBD) currently relies on aspecific clinical signs of bowel inflammation. Specific imaging of the diseased bowel regions is still lacking. Here, we investigate mucosal addressin cell adhesion molecule 1 (MAdCAM-1) as a reliable and specific endothelial target for engineered nanoparticles delivering imaging agents to obtain an exact mapping of diseased bowel foci. MATERIALS AND METHODS: We generated a nanodevice composed of PLGA-PEG coupled with anti-MAdCAM-1 antibody half-chains and loaded with quantum dots (P@QD-MdC NPs). Bowel localization and systemic biodistribution of the nanoconjugate were analyzed upon injection in a murine model of chronic IBD obtained through repeated administration of dextran sulfate sodium salt. Specificity for diseased bowel regions was also assessed ex vivo in human specimens from patients with IBD. Potential for development as contrast agent in magnetic resonance imaging was assessed by preliminary study on animal model. RESULTS: Synthesized nanoparticles revealed good stability and monodispersity. Molecular targeting properties were analyzed in vitro in a cell culture model. Upon intravenous injection, P@QD-MdC NPs were localized in the bowel of colitic mice, with enhanced accumulation at 24 h post-injection compared to untargeted nanoparticles (p<0.05). Nanoparticles injection did not induce histologic lesions in non-target organs. Ex vivo exposure of human bowel specimens to P@QD-MdC NPs revealed specific recognition of the diseased regions vs uninvolved tracts (p<0.0001). After loading with appropriate contrast agent, the nanoparticles enabled localized contrast enhancement of bowel mucosa in the rectum of treated mice. CONCLUSION: P@QD-MdC NPs efficiently detected bowel inflammation foci, accurately following the expression pattern of MAdCAM-1. Fine-tuning of this nanoconjugate with appropriate imaging agents offers a promising non-invasive tool for specific IBD diagnosis.
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Moléculas de Adesão Celular/imunologia , Imunoconjugados/administração & dosagem , Doenças Inflamatórias Intestinais/diagnóstico por imagem , Mucoproteínas/imunologia , Pontos Quânticos/administração & dosagem , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/imunologia , Colite/induzido quimicamente , Colite/diagnóstico por imagem , Doença de Crohn/diagnóstico por imagem , Modelos Animais de Doenças , Feminino , Humanos , Imunoconjugados/farmacocinética , Injeções Intravenosas , Mucosa Intestinal/diagnóstico por imagem , Intestinos/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Camundongos Endogâmicos C57BL , Nanopartículas/administração & dosagem , Nanopartículas/química , Poliésteres/química , Polietilenoglicóis/química , Distribuição TecidualRESUMO
Dysfunctional epicardial adipose tissue (EAT) secretome can influence the heart's stretch response. However, the molecular mechanisms are still poorly understood. The aim of this study was to clarify how dysfunctional EAT promotes maladaptive heart remodeling in cardiovascular disease (CVD) through ST2 production associated with exchange protein directly activated by cAMP (EPAC) proteins. A series of 55 CVD males were enrolled and their EAT thickness, LV mass and volumes were measured by echocardiography. Blood, plasma and EAT biopsies were collected for molecular and proteomic assays. Taking EAT thickness as a continuous variable there was a direct correlation between the ST2 cardiac stretch mediator and EAT thickness (r = 0.54, p < 0.01) and an inverse relation between the ST2 gene and IL-33 expression (r -0.50, p < 0.01). In the CVD population EPAC2 expression directly correlated with the ST2 gene (r = 0.74, p < 0.0001) causing an ST2/IL-33 system local (p < 0.001) and systemic (sST2 = 57.33 ± 3.22 and IL-33 = 0.53 ± 017 pg/mL; p < 0.0001) protein imbalance associated with maladaptive remodeling. This indicated that dysfunctional EAT is a source of both EPAC and ST2 protein and an EPAC2 isoform seems involved in ST2 production in adipose tissue. Both EPAC2 and ST2 expression were directly related to maladaptive heart remodeling indices, suggesting EAT measurements could be useful in the early assessment of CVD complications.
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Doenças Cardiovasculares/metabolismo , Doenças Cardiovasculares/patologia , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Proteína 1 Semelhante a Receptor de Interleucina-1/metabolismo , Interleucina-33/metabolismo , Gordura Intra-Abdominal/metabolismo , Gordura Intra-Abdominal/patologia , Pericárdio/metabolismo , Pericárdio/patologia , Remodelação Ventricular/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , Doenças Cardiovasculares/diagnóstico por imagem , Ecocardiografia , Humanos , Hipertrofia Ventricular Esquerda/diagnóstico por imagem , Hipertrofia Ventricular Esquerda/metabolismo , Hipertrofia Ventricular Esquerda/patologia , Gordura Intra-Abdominal/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Pericárdio/diagnóstico por imagem , Transdução de SinaisRESUMO
OBJECTIVE: To study pathogenic features of the somatic testicular microenvironment associated with idiopathic germ cell aplasia. DESIGN: Cross-sectional study. SETTING: Tertiary referral center for reproductive medicine. PATIENT(S): Testicular specimens from men with idiopathic nonobstructive azoospermia (iNOA) prospectively submitted to microdissection testicular sperm extraction. Of 20 specimens used for histology, 10 were also available for proteomic analysis. Primary Sertoli cells with normal karyotype and phenotype were also used. INTERVENTION(S): Patients with iNOA were dichotomized according to a positive versus negative sperm retrieval at microdissection testicular sperm extraction, and on the isolated extracellular matrix (ECM) the proteomic analysis was performed. MAIN OUTCOME MEASURE(S): Proteomic analysis of the ECM from testicular specimens with positive versus negative sperm retrieval. Gene ontology enrichment was used to identify upstream regulators based on the 11 deregulated ECM proteins, which were validated by immunohistochemistry and quantitative polymerase chain reaction. Continuous variables were expressed as medians and interquartile range. RESULT(S): Germ cell aplasia was characterized by an increased signaling of the retinoic acid in Sertoli cells and associated with decreased expression of the basal membrane markers nidogen-2 and heparan sulfate proteoglycan-2. Decreased levels of the interstitial matrisome-associated factor IX and its regulator VKORC1 were, instead, coupled with decreased signaling of vitamin K in Leydig cells. An altered expression of a further eight ECM proteins was also found, including laminin-4 and laminin-5. Peripheral levels of the two vitamins were within the reference range in the two cohorts of iNOA men. CONCLUSION(S): We identified the pathogenetic signature of the somatic human testicular microenvironment, providing two vitamin-related mechanistic insights related to the molecular determinants of the idiopathic germ cell aplasia.
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Azoospermia/metabolismo , Azoospermia/patologia , Matriz Extracelular/patologia , Testículo/metabolismo , Vitamina A/metabolismo , Vitamina K/metabolismo , Adolescente , Células Cultivadas , Estudos Transversais , Matriz Extracelular/metabolismo , Humanos , Masculino , Microdissecção , Proteômica , Transdução de Sinais/fisiologia , Recuperação Espermática , Testículo/patologia , Adulto JovemRESUMO
One of the most important Staphylococcus aureus virulence factors is Panton-Valentine leukocidin (PVL). We describe an outbreak of recurrent cutaneous PVL infections in different members of three family clusters. Molecular investigations were performed to confirm the presence of the mecA and PVL genes and to assign the SCCmec type, sequence type (ST) and clonal relatedness. A strain of PVL-producing methicillin-resistant S. aureus (MRSA) was responsible for infection in two related families (A and B), and a third family (C) was infected with PVL-producing methicillin-sensitive S. aureus (MSSA). Molecular investigations revealed the same clone of community-acquired (CA)-MRSA, PVL positive ST8, and SCCmec IV in families A and B and CA-MSSA PVL positive ST15 in family C. S. aureus PVL may give rise to recurrent uncontrolled infections that are difficult to eradicate, and close family contacts are at high risk for transmission.
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Infecções Comunitárias Adquiridas/epidemiologia , Infecções Estafilocócicas/epidemiologia , Staphylococcus aureus/isolamento & purificação , Abscesso/microbiologia , Adulto , Toxinas Bacterianas/biossíntese , Criança , Infecções Comunitárias Adquiridas/microbiologia , Exotoxinas/biossíntese , Feminino , Humanos , Lactente , Leucocidinas/biossíntese , Masculino , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/fisiologia , Fatores de VirulênciaRESUMO
Human parvovirus 4 (PARV4) is a recently discovered member of the Parvoviridae. We investigated the presence of this virus in bone-marrow aspirates of 35 Italian patients with AIDS. Viral DNA was detected by polymerase chain reaction in over 40% of patients (16/35). The infection was most prevalent in injection drug users (IDU; 12/18; 66.7%) as opposed to non-IDU (4/17; 23.5%). PARV4 infection is widespread in Italian patients with AIDS.
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Síndrome da Imunodeficiência Adquirida/virologia , Medula Óssea/virologia , DNA Viral/análise , HIV-1 , Infecções por Parvoviridae/virologia , Parvoviridae/genética , Adulto , Feminino , Genótipo , Hepacivirus/genética , Hepatite C/virologia , Humanos , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Prevalência , Abuso de Substâncias por Via Intravenosa/virologia , Carga ViralRESUMO
BACKGROUND: To overcome some of the limitations of conventional microbiologic techniques, polymerase chain reaction (PCR)-based assays are proposed as useful tools for the diagnosis of visceral leishmaniasis. PATIENTS AND METHODS: A comparative study using conventional microbiologic techniques (i.e., serologic testing, microscopic examination, and culture) and a Leishmania species-specific PCR assay, using peripheral blood and bone marrow aspirate samples as templates, was conducted during an 8-year period. The study cohort consisted of 594 Italian immunocompetent (adult and pediatric) and immunocompromised (adult) patients experiencing febrile syndromes associated with hematologic alterations and/or hepatosplenomegaly. Identification of the infecting protozoa at the species level was directly obtained by PCR of peripheral blood samples, followed by restriction fragment-length polymorphism analysis of the amplified products, and the results were compared with those of isoenzyme typing of Leishmania species strains from patients, which were isolated in vitro. RESULTS: Sixty-eight patients (11.4%) had a confirmed diagnosis of visceral leishmaniasis. Eleven cases were observed in human immunodeficiency virus (HIV)-uninfected adults, 20 cases were observed in HIV-infected adults, and the remaining 37 cases were diagnosed in HIV-uninfected children. In the diagnosis of primary visceral leishmaniasis, the sensitivities of the Leishmania species-specific PCR were 95.7% for bone marrow aspirate samples and 98.5% for peripheral blood samples versus sensitivities of 76.2%, 85.5%, and 90.2% for bone marrow aspirate isolation, serologic testing, and microscopic examination of bone marrow biopsy specimens, respectively. None of 229 healthy blood donors or 25 patients with imported malaria who were used as negative control subjects had PCR results positive for Leishmania species in peripheral blood samples (i.e., specificity of Leishmania species-specific PCR, 100%). PCR and restriction fragment-length polymorphism analysis for Leishmania species identification revealed 100% concordance with isoenzyme typing in the 19 patients for whom the latter data were available. CONCLUSIONS: PCR assay is a highly sensitive and specific tool for the diagnosis of visceral leishmaniasis in both immunocompetent and immunocompromised patients and can be reliably used for rapid parasite identification at the species level.
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Medula Óssea/parasitologia , Hospedeiro Imunocomprometido , Leishmania/genética , Leishmaniose Visceral/diagnóstico , Reação em Cadeia da Polimerase/métodos , Testes Sorológicos/métodos , Infecções Oportunistas Relacionadas com a AIDS , Adulto , Idoso , Algoritmos , Animais , Criança , Pré-Escolar , Feminino , Infecções por HIV/complicações , Humanos , Lactente , Itália , Leishmania/classificação , Leishmania/isolamento & purificação , Leishmaniose Visceral/sangue , Leishmaniose Visceral/genética , Leishmaniose Visceral/virologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Sensibilidade e EspecificidadeRESUMO
AIM: We investigate MAdCAM-1 as a reliable target to detect active bowel inflammation for selective noninvasive nanodiagnostics. MATERIALS & METHODS: We coupled anti-MAdCAM-1 antibodies to manganese oxide nanoparticles, and analyzed nanoconjugate biodistribution and safety in murine model of inflammatory bowel disease by imaging and histology. RESULTS: Nanoparticles were stable and nontoxic. Upon administration in colitic mice, anti-MAdCAM-1 functionalized nanoparticles preferentially localized in the inflamed bowel, whereas untargeted nanoparticles were more rapidly washed out. Nanoparticles did not induce lesions in nontarget organs. CONCLUSION: Anti-MAdCAM-1 functionalized nanoparticles detected active bowel inflammation foci, accurately following MAdCAM-1 expression pattern. These nanoconjugates could be a promising noninvasive imaging system for an early and accurate follow-up in patients affected by acute colitis.
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Anticorpos/química , Colite/diagnóstico por imagem , Imunoglobulinas/imunologia , Compostos de Manganês/química , Nanopartículas Metálicas/química , Mucoproteínas/imunologia , Óxidos/química , Animais , Moléculas de Adesão Celular , Sobrevivência Celular , Colite/metabolismo , Colite/patologia , Hemólise , Humanos , Imunoglobulinas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mucoproteínas/metabolismo , Tamanho da Partícula , Propriedades de Superfície , Distribuição TecidualRESUMO
Chemotherapeutic treatment of breast cancer is based on maximum tolerated dose (MTD) approach. However, advanced stage tumors are not effectively eradicated by MTD owing to suboptimal drug targeting, onset of therapeutic resistance and neoangiogenesis. In contrast, "metronomic" chemotherapy is based on frequent drug administrations at lower doses, resulting in neovascularization inhibition and induction of tumor dormancy. Here we show the potential of H-ferritin (HFn)-mediated targeted nanodelivery of metronomic doxorubicin (DOX) in the setting of a highly aggressive and metastatic 4T1 breast cancer mouse model with DOX-inducible expression of chemoresistance. We find that HFn-DOX administered at repeated doses of 1.24 mg kg-1 strongly improves the antitumor potential of DOX chemotherapy arresting the tumor progression. We find that such a potent antitumor effect is attributable to multiple nanodrug actions beyond cell killing, including inhibition of tumor angiogenesis and avoidance of chemoresistance. Multiparametric assessment of heart tissues, including histology, ultrastructural analysis of tissue morphology, and measurement of markers of reactive oxygen species and hepatic/renal conditions, provided evidence that metronomic HFn-DOX allowed us to overcome cardiotoxicity. Our results suggest that HFn-DOX has tremendous potential for the development of "nanometronomic" chemotherapy toward safe and tailored oncological treatments.
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Antibióticos Antineoplásicos/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , Doxorrubicina/administração & dosagem , Resistencia a Medicamentos Antineoplásicos , Cardiopatias/prevenção & controle , Nanomedicina/métodos , Nanopartículas , Administração Metronômica , Animais , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/farmacocinética , Antibióticos Antineoplásicos/toxicidade , Apoferritinas/química , Apoferritinas/metabolismo , Disponibilidade Biológica , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Cardiotoxicidade , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/química , Doxorrubicina/farmacocinética , Doxorrubicina/toxicidade , Portadores de Fármacos , Composição de Medicamentos , Feminino , Cardiopatias/induzido quimicamente , Camundongos Endogâmicos BALB C , Neovascularização Patológica , Distribuição Tecidual , Carga Tumoral/efeitos dos fármacosRESUMO
BACKGROUND: Clinical experience highlights the wide heterogeneity of primary prostate cancer (PPCa), even when potentially related to the same grade and stage. Currently available prediction tools and biomarkers do not always allow for early recognition of PPCa aggressive phenotype, sometimes making it impossible to distinguish among men harbouring indolent tumours or life-threatening disease. OBJECTIVE: To establish a novel ex vivo/in vitro model suitable to estimate the invasive phenotype of PPCa cells (PPCaC). DESIGN, SETTING, AND PARTICIPANTS: The ability of PPCaC to infiltrate the prostate extracellular matrix (ECM) was used as an index of invasion. ECM was obtained by decellularising 24 NT-prostate specimens from radical prostatectomy. PPCaC were obtained from six tumours with different Gleason patterns and pathological stages. Invasion ability was estimated in direct-cocolture experiments. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: The extent of ECM invasion by PPCaC was quantified by counting the number of infiltrated cells. Mann-Whitney test was utilised for statistical comparisons. RESULTS AND LIMITATIONS: Samples of ECM resulted to be free of cells and DNA and with a preserved three-dimensional structure and stromal protein content. The system resulted to be reliable since well characterised normal-, benign-, and malignant-prostate cell lines either re-epitheliased or invaded the matrices, according to their specific nature. Similarly, PPCaC invaded the ECMs consistently with their stage and biochemical recurrence. Of notice, this model was able to identify a different invasive phenotype even among tumours with equal Gleason patterns and pathological stages. The small sample size represents a limitation. CONCLUSIONS: We developed an ex vivo/in vitro model able to reproduce the original PPCa-microenvironment and suitable to recognise the inherent invasive behaviour of PPCaC. PATIENT SUMMARY: We developed a novel ex vivo/in vitro system which enables us to uncover which prostate tumours host potentially aggressive cancer cells. The identification of cancer cells with different invasive abilities will likely lead to the identification of new biomarkers to safely predict disease progression.
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The extracellular matrix (ECM) from perilesional and colorectal carcinoma (CRC), but not healthy colon, sustains proliferation and invasion of tumor cells. We investigated the biochemical and physical diversity of ECM in pair-wised comparisons of healthy, perilesional and CRC specimens. Progressive linearization and degree of organization of fibrils was observed from healthy to perilesional and CRC ECM, and was associated with a steady increase of stiffness and collagen crosslinking. In the perilesional ECM these modifications coincided with increased vascularization, whereas in the neoplastic ECM they were associated with altered modulation of matrisome proteins, increased content of hydroxylated lysine and lysyl oxidase. This study identifies the increased stiffness and crosslinking of the perilesional ECM predisposing an environment suitable for CRC invasion as a phenomenon associated with vascularization. The increased stiffness of colon areas may represent a new predictive marker of desmoplastic region predisposing to invasion, thus offering new potential application for monitoring adenoma with invasive potential.
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Colo/metabolismo , Neoplasias Colorretais/metabolismo , Matriz Extracelular/metabolismo , Proteína-Lisina 6-Oxidase/metabolismo , Idoso , Idoso de 80 Anos ou mais , Movimento Celular , Colágeno/metabolismo , Colo/patologia , Neoplasias Colorretais/irrigação sanguínea , Neoplasias Colorretais/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Neovascularização Patológica , Proteína-Lisina 6-Oxidase/genética , Microambiente TumoralRESUMO
In the fields of biomaterials and tissue engineering simulating the native microenvironment is of utmost importance. As a major component of the microenvironment, the extracellular matrix (ECM) contributes to tissue homeostasis, whereas modifications of native features are associated with pathological conditions. Furthermore, three-dimensional (3D) geometry is an important feature of synthetic scaffolds favoring cell stemness, maintenance and differentiation. We analyzed the 3D structure, geometrical measurements and anisotropy of the ECM isolated from (i) human bladder mucosa (basal lamina and lamina propria) and muscularis propria; and, (ii) bladder carcinoma (BC). Next, binding and invasion of bladder metastatic cell line was observed on synthetic scaffold recapitulating anisotropy of tumoral ECM, but not on scaffold with disorganized texture typical of non-neoplastic lamina propria. This study provided information regarding the ultrastructure and geometry of healthy human bladder and BC ECMs. Likewise, using synthetic scaffolds we identified linearization of the texture as a mandatory feature for BC cell invasion. Integrating microstructure and geometry with biochemical and mechanical factors could support the development of an innovative synthetic bladder substitute or a tumoral scaffold predictive of chemotherapy outcomes.
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Matriz Extracelular/patologia , Invasividade Neoplásica/patologia , Microambiente Tumoral , Neoplasias da Bexiga Urinária/patologia , Idoso , Idoso de 80 Anos ou mais , Histocitoquímica , Humanos , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Mucosa/patologia , Bexiga Urinária/patologiaRESUMO
Merkel cell carcinoma (MCC) is a skin cancer that can also rarely arise in extracutaneous sites including mucosal surfaces. About 80% of MCCs harbor the Merkel cell polyomavirus (MCPyV). All cases of gastric MCCs so far reported were metastases from cutaneous sources. In the present article, we describe for the first time a primary gastric MCC harboring MCPyV. A 72-year-old man presented to clinical observation due to epigastric pain. Upper endoscopy revealed an ulcerated gastric tumor. The patient underwent total gastrectomy. The tumor was composed of mitotically active monomorphic small cells showing round nuclei with finely dispersed chromatin arranged in sheets and nests with large areas of necrosis. Tumor cells were positive for neuroendocrine markers and showed paranuclear dot immunoreactivity for cytokeratin 20. MCPyV was demonstrated with immunohistochemistry and electron microscopy, which showed intranuclear and intracytoplasmic viral particles. The MCPyV DNA in tumor cells was demonstrated with polymerase chain reaction analysis.
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Carcinoma de Célula de Merkel/patologia , Carcinoma de Célula de Merkel/virologia , Infecções por Polyomavirus/virologia , Neoplasias Gástricas/patologia , Neoplasias Gástricas/virologia , Infecções Tumorais por Vírus/virologia , Idoso , Carcinoma Neuroendócrino , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Masculino , Poliomavírus das Células de Merkel , Reação em Cadeia da Polimerase Multiplex , Gradação de Tumores , Infecções por Polyomavirus/patologia , Infecções Tumorais por Vírus/patologiaRESUMO
AIM: Immunohistochemical and molecular studies have suggested an oncogenic role for JCV in gastrointestinal carcinomas, but at least in colorectal cancers, the data are far from being unambiguous. METHODS: Two large series of formalin-fixed paraffin-embedded gastric and colorectal cancers were analysed for the expression of JCV large T Antigen (T-Ag) with a panel of five antibodies, and for the presence of T-Ag DNA sequences using two PCR systems. RESULTS: Intense nuclear staining was observed in 54/116 (46%) colorectal, and in 92/234 (39%) gastric cancers, using the PAb416 monoclonal antibody against large T-Ag. In colorectal cancers, PAb416-positivity was directly related to the presence of chromosomal instability, lymph node metastases and a more advanced tumour stage, and inversely related to proximal tumour site and the presence of microsatellite instability (MSI). In gastric cancers, the glandular histotype, the presence of lymph node metastases, a low frequency of MSI and EBV infection, and a worse prognosis were significantly associated with PAb416 immunoreactivity. Moreover, at both these sites, PAb416 expression was significantly associated with p53 nuclear accumulation. No positivity was obtained with all the other four anti-T-Ag-antibodies, and molecular analysis failed to demonstrate the presence of JCV DNA sequences in tested cases. CONCLUSIONS: Our immunohistochemical and molecular results do not support the idea that JCV T-Ag has a role in gastrointestinal carcinogenesis. It is possible that PAb416, besides binding the viral protein, may cross-react with a hitherto undefined protein whose expression is associated with a distinct pathological profile and, at least in gastric cancers, with worse prognosis.
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Adenocarcinoma/virologia , Antígenos Transformantes de Poliomavirus/imunologia , Neoplasias Gastrointestinais/virologia , Vírus JC/genética , Adenocarcinoma/genética , Adenocarcinoma/secundário , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais , Western Blotting , Núcleo Celular/metabolismo , Núcleo Celular/patologia , Reações Cruzadas , DNA Viral/análise , Feminino , Neoplasias Gastrointestinais/genética , Neoplasias Gastrointestinais/patologia , Humanos , Vírus JC/imunologia , Linfonodos/patologia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNAAssuntos
Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Mioclonia/diagnóstico por imagem , Mioclonia/patologia , Doença de Whipple/diagnóstico por imagem , Doença de Whipple/patologia , Encéfalo/fisiopatologia , Diagnóstico Diferencial , Evolução Fatal , Humanos , Masculino , Pessoa de Meia-Idade , Mioclonia/fisiopatologia , Mioclonia/terapia , Doença de Whipple/fisiopatologia , Doença de Whipple/terapiaRESUMO
OBJECTIVES: Low 25-Hydroxyvitamin D (25[OH]D) was associated with severe fibrosis and low sustained virological response (SVR) after interferon (IFN)-based therapy in chronic hepatitis C. Furthermore, hypovitaminosis D was reported in HIV-infected individuals, but its role in liver disease progression in HIV/HCV coinfection is unknown. METHODS: 25(OH)D was retrospectively measured in 237 HIV-infected patients (93 with HCV coinfection) and 76 healthy controls. Multivariate analysis included season, immuno-virological data, combined antiretroviral therapy (cART) and, in a subgroup of 51 HIV/HCV-genotype 1 coinfected patients, factors influencing SVR to pegylated-IFN and ribavirin. In a group of 20 patients, liver expression of cytochrome (CY)-P27A1 and CYP2R1, 25-hydroxylating enzymes, was assessed by immunohistochemistry. RESULTS: Median 25(OH)D levels were 23.4 (interquartile range 16.7-33.7) ng/mL in the HIV-infected population and 24 ng/mL (18.3-29.5) in healthy controls (p=0.9). At multiple regression analysis, only winter/spring measurements correlated with lower 25(OH)D levels. No correlation with HCV coinfection, nor with cART regimens was found. Low 25(OH)D was independently associated with advanced fibrosis in HIV/HCV coinfected patients (p=0.023), whereas no association emerged with SVR to IFN-based therapy. CYP27A1 and CYP2R1 expression was associated neither with 25(OH)D serum levels nor with HCV-infection, liver histology, or cART. CONCLUSIONS: In our experience, despite the high prevalence of 25(OH)D insufficiency, HIV and HCV-infection did not seem to influence vitamin D status. The role of HIV, HCV and cART on hypovitaminosis D needs further validation in larger cohorts that account for the vitamin levels in general populations and for seasonal and regional variability.