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1.
FASEB J ; 35(10): e21897, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34473378

RESUMO

Autosomal dominant polycystic kidney disease (ADPKD) is characterized by the development of bilateral renal cysts which enlarge continuously, leading to compression of adjacent intact nephrons. The growing cysts lead to a progressive decline in renal function. Cyst growth is driven by enhanced cell proliferation and chloride secretion into the cyst lumen. Chloride secretion is believed to occur mainly by the cAMP-activated cystic fibrosis transmembrane conductance regulator (CFTR), with some contribution by the calcium-activated chloride channel TMEM16A. However, our previous work suggested TMEM16A as a major factor for renal cyst formation. The contribution of CFTR to cyst formation has never been demonstrated in an adult ADPKD mouse model. We used mice with an inducible tubule-specific Pkd1 knockout, which consistently develop polycystic kidneys upon deletion of Pkd1. Cellular properties, ion currents, and cyst development in these mice were compared with that of mice carrying a co-deletion of Pkd1 and Cftr. Knockout of Cftr did not reveal any significant impact on cyst formation in the ADPKD mouse model. Furthermore, knockout of Cftr did not attenuate the largely augmented cell proliferation observed in Pkd1 knockout kidneys. Patch clamp analysis on primary renal epithelial cells lacking expression of Pkd1 indicated an only marginal contribution of CFTR to whole cell Cl- currents, which were clearly dominated by calcium-activated TMEM16A currents. In conclusion, CFTR does not essentially contribute to renal cyst formation in mice caused by deletion of Pkd1. Enhanced cell proliferation and chloride secretion is caused primarily by upregulation of the calcium-activated chloride channel TMEM16A.


Assuntos
Anoctamina-1/metabolismo , Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Cistos/metabolismo , Rim Policístico Autossômico Dominante/metabolismo , Canais de Cátion TRPP/metabolismo , Animais , Anoctamina-1/genética , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Cistos/genética , Cistos/patologia , Modelos Animais de Doenças , Técnicas de Silenciamento de Genes , Camundongos , Camundongos Knockout , Rim Policístico Autossômico Dominante/genética , Rim Policístico Autossômico Dominante/patologia , Canais de Cátion TRPP/genética
2.
FASEB J ; 35(10): e21865, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34486178

RESUMO

Autosomal dominant polycystic kidney disease is a common inherited renal disorder that results from mutations in either PKD1 or PKD2, encoding polycystin-1 (PC1) and polycystin-2 (PC2), respectively. Downregulation or overexpression of PKD1 or PKD2 in mouse models results in renal cyst formation, suggesting that the quantity of PC1 and PC2 needs to be maintained within a tight functional window to prevent cystogenesis. Here we show that enhanced PC2 expression is a common feature of PKD1 mutant tissues, in part due to an increase in Pkd2 mRNA. However, our data also suggest that more effective protein folding contributes to the augmented levels of PC2. We demonstrate that the unfolded protein response is activated in Pkd1 knockout kidneys and in Pkd1 mutant cells and that this is coupled with increased levels of GRP94, an endoplasmic reticulum protein that is a member of the HSP90 family of chaperones. GRP94 was found to physically interact with PC2 and depletion or chemical inhibition of GRP94 led to a decrease in PC2, suggesting that GRP94 serves as its chaperone. Moreover, GRP94 is acetylated and binds to histone deacetylase 6 (HDAC6), a known deacetylase and activator of HSP90 proteins. Inhibition of HDAC6 decreased PC2 suggesting that HDAC6 and GRP94 work together to regulate PC2 levels. Lastly, we showed that inhibition of GRP94 prevents cAMP-induced cyst formation in vitro. Taken together our data uncovered a novel HDAC6-GRP94-related axis that likely participates in maintaining elevated PC2 levels in Pkd1 mutant cells.


Assuntos
Cistos/patologia , Retículo Endoplasmático/metabolismo , Nefropatias/patologia , Glicoproteínas de Membrana/metabolismo , Fator de Transcrição PAX8/fisiologia , Canais de Cátion TRPP/fisiologia , Animais , Cálcio/metabolismo , Cistos/etiologia , Cistos/metabolismo , Nefropatias/etiologia , Nefropatias/metabolismo , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Resposta a Proteínas não Dobradas
3.
Nat Commun ; 12(1): 4548, 2021 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-34315885

RESUMO

Autosomal dominant polycystic kidney disease (ADPKD) is caused by germline mutations of PKD1 or PKD2 on one allele and a somatic mutation inactivating the remaining normal allele. However, if and how null ADPKD gene renal epithelial cells affect the biology and function of neighboring cells, including heterozygous renal epithelial cells, fibroblasts and macrophages during cyst initiation and expansion remains unknown. Here we address this question with a "cystic extracellular vesicles/exosomes theory". We show that cystic cell derived extracellular vesicles and urinary exosomes derived from ADPKD patients promote cyst growth in Pkd1 mutant kidneys and in 3D cultures. This is achieved by: 1) downregulation of Pkd1 gene expression and upregulation of specific miRNAs, resulting in the activation of PKD associated signaling pathways in recipient renal epithelial cells and tissues; 2) the activation of fibroblasts; and 3) the induction of cytokine expression and the recruitment of macrophages to increase renal inflammation in cystic kidneys. Inhibition of exosome biogenesis/release with GW4869 significantly delays cyst growth in aggressive and milder ADPKD mouse models, suggesting that targeting exosome secretion has therapeutic potential for ADPKD.


Assuntos
Células Epiteliais/patologia , Exossomos/metabolismo , Rim/patologia , Rim Policístico Autossômico Dominante/patologia , Compostos de Anilina/farmacologia , Animais , Compostos de Benzilideno/farmacologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Colágeno , Cistos/patologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Exossomos/efeitos dos fármacos , Fibrose , Regulação da Expressão Gênica/efeitos dos fármacos , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , Modelos Biológicos , Mutação/genética , Rim Policístico Autossômico Dominante/genética , Rim Policístico Autossômico Dominante/urina , Ratos , Transdução de Sinais/efeitos dos fármacos , Canais de Cátion TRPP/metabolismo , Tetraspanina 30/metabolismo
4.
Ann Agric Environ Med ; 28(2): 348-351, 2021 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-34184522

RESUMO

Introduction. Epididymal dirofilariasis is one of the unusual manifestations of this zoonosis. In Slovakia, this is the third case out of 20 Dirofilaria repens infected patients in whom the worm (the parasite) was identified in the epididymis. The patient felt a painless tumour about 2 cm in size on the left testicle. During ultrasound examination, the radiologist observed a cyst containing a live worm within the epididymis. After mechanical stimulation, the movement characteristic for filarial worms (´filarial dance sign´) appeared. An orchiectomy was performed at the Department of Urology in the University Hospital in Kosice. Histopathology confirmed a parasitic cyst with a worm, and based on characteristic morphological features, the parasite was identified as Dirofilaria repens. Objective. The aim of case report is to alert physicians to the possibility dirofilarial infection of the epididymis, where tumors and cystic structures often occur and to present clinical signs of the disease.


Assuntos
Cistos/parasitologia , Dirofilaria repens/isolamento & purificação , Dirofilariose/parasitologia , Epididimo/parasitologia , Doenças dos Genitais Masculinos/parasitologia , Idoso , Animais , Cistos/patologia , Dirofilaria repens/classificação , Dirofilaria repens/genética , Dirofilariose/patologia , Epididimo/patologia , Doenças dos Genitais Masculinos/patologia , Humanos , Masculino , Eslováquia
5.
J Am Soc Nephrol ; 32(8): 1913-1932, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34155062

RESUMO

BACKGROUND: In autosomal dominant polycystic kidney disease (ADPKD), cyst development and enlargement lead to ESKD. Macrophage recruitment and interstitial inflammation promote cyst growth. TWEAK is a TNF superfamily (TNFSF) cytokine that regulates inflammatory responses, cell proliferation, and cell death, and its receptor Fn14 (TNFRSF12a) is expressed in macrophage and nephron epithelia. METHODS: To evaluate the role of the TWEAK signaling pathway in cystic disease, we evaluated Fn14 expression in human and in an orthologous murine model of ADPKD. We also explored the cystic response to TWEAK signaling pathway activation and inhibition by peritoneal injection. RESULTS: Meta-analysis of published animal-model data of cystic disease reveals mRNA upregulation of several components of the TWEAK signaling pathway. We also observed that TWEAK and Fn14 were overexpressed in mouse ADPKD kidney cysts, and TWEAK was significantly high in urine and cystic fluid from patients with ADPKD. TWEAK administration induced cystogenesis and increased cystic growth, worsening the phenotype in a murine ADPKD model. Anti-TWEAK antibodies significantly slowed the progression of ADPKD, preserved renal function, and improved survival. Furthermore, the anti-TWEAK cystogenesis reduction is related to decreased cell proliferation-related MAPK signaling, decreased NF-κB pathway activation, a slight reduction of fibrosis and apoptosis, and an indirect decrease in macrophage recruitment. CONCLUSIONS: This study identifies the TWEAK signaling pathway as a new disease mechanism involved in cystogenesis and cystic growth and may lead to a new therapeutic approach in ADPKD.


Assuntos
Citocina TWEAK/metabolismo , Rim Policístico Autossômico Dominante/metabolismo , Rim Policístico Autossômico Dominante/patologia , Receptor de TWEAK/metabolismo , Adulto , Animais , Anticorpos Neutralizantes/farmacologia , Apoptose , Proliferação de Células/efeitos dos fármacos , Cistos/metabolismo , Cistos/patologia , Citocina TWEAK/antagonistas & inibidores , Citocina TWEAK/genética , Citocina TWEAK/farmacologia , Modelos Animais de Doenças , Progressão da Doença , Feminino , Fibrose , Expressão Gênica , Humanos , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos , Masculino , Camundongos , Pessoa de Meia-Idade , NF-kappa B/metabolismo , Rim Policístico Autossômico Dominante/fisiopatologia , Transdução de Sinais , Receptor de TWEAK/genética
6.
PLoS One ; 16(4): e0248400, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33886581

RESUMO

Augmentation of endogenous nitric oxide (NO) synthesis, either by the classical L-arginine-NO synthase pathway, or the recently discovered entero-salivary nitrate-nitrite-NO system, may slow the progression of autosomal dominant polycystic kidney disease (ADPKD). To test this hypothesis, the expression of NO in human ADPKD cell lines (WT 9-7, WT 9-12), and the effect of L-arginine on an in vitro model of three-dimensional cyst growth using MDCK cells, was examined. In addition, groups of homozygous Pkd1RC/RC mice (a hypomorphic genetic ortholog of ADPKD) received either low, moderate or high dose sodium nitrate (0.1, 1 or 10 mmol/kg/day), or sodium chloride (vehicle; 10 mmol/kg/day), supplemented drinking water from postnatal month 1 to 9 (n = 12 per group). In vitro, intracellular NO, as assessed by DAF-2/DA fluorescence, was reduced by >70% in human ADPKD cell lines, and L-arginine and the NO donor, sodium nitroprusside, both attenuated in vitro cyst growth by up to 18%. In contrast, in Pkd1RC/RC mice, sodium nitrate supplementation increased serum nitrate/nitrite levels by ~25-fold in the high dose group (P<0.001), but kidney enlargement and percentage cyst area was not altered, regardless of dose. In conclusion, L-arginine has mild direct efficacy on reducing renal cyst growth in vitro, whereas long-term sodium nitrate supplementation was ineffective in vivo. These data suggest that the bioconversion of dietary nitrate to NO by the entero-salivary pathway may not be sufficient to influence the progression of renal cyst growth in ADPKD.


Assuntos
Suplementos Nutricionais , Rim/patologia , Nitratos/uso terapêutico , Rim Policístico Autossômico Dominante/patologia , Rim Policístico Autossômico Dominante/terapia , Animais , Linhagem Celular , Cistos/patologia , Cistos/terapia , Cães , Feminino , Humanos , Células Madin Darby de Rim Canino , Masculino , Camundongos , Camundongos Endogâmicos C57BL
7.
FASEB J ; 35(5): e21533, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33826787

RESUMO

Polycystic kidney disease (PKD) is a genetic disorder characterized by aberrant renal epithelial cell proliferation and formation and progressive growth of numerous fluid-filled cysts within the kidneys. Previously, we showed that there is elevated Notch signaling compared to normal renal epithelial cells and that Notch signaling contributes to the proliferation of cystic cells. Quinomycin A, a bis-intercalator peptide, has previously been shown to target the Notch signaling pathway and inhibit tumor growth in cancer. Here, we show that Quinomycin A decreased cell proliferation and cyst growth of human ADPKD cyst epithelial cells cultured within a 3D collagen gel. Treatment with Quinomycin A reduced kidney weight to body weight ratio and decreased renal cystic area and fibrosis in Pkd1RC/RC ; Pkd2+/- mice, an orthologous PKD mouse model. This was accompanied by reduced expression of Notch pathway proteins, RBPjk and HeyL and cell proliferation in kidneys of PKD mice. Quinomycin A treatments also normalized cilia length of cyst epithelial cells derived from the collecting ducts. This is the first study to demonstrate that Quinomycin A effectively inhibits PKD progression and suggests that Quinomycin A has potential therapeutic value for PKD patients.


Assuntos
Antibacterianos/farmacologia , Cistos/tratamento farmacológico , Modelos Animais de Doenças , Equinomicina/farmacologia , Doenças Renais Policísticas/complicações , Canais de Cátion TRPP/fisiologia , Animais , Cistos/etiologia , Cistos/metabolismo , Cistos/patologia , Progressão da Doença , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout
8.
Am J Physiol Lung Cell Mol Physiol ; 320(6): L1158-L1168, 2021 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-33881909

RESUMO

The TGF-ß signaling pathway plays a pivotal role in controlling organogenesis during fetal development. Although the role of TGF-ß signaling in promoting lung alveolar epithelial growth has been determined, mesenchymal TGF-ß signaling in regulating lung development has not been studied in vivo due to a lack of genetic tools for specifically manipulating gene expression in lung mesenchymal cells. Therefore, the integral roles of TGF-ß signaling in regulating lung development and congenital lung diseases are not completely understood. Using a Tbx4 lung enhancer-driven Tet-On inducible Cre transgenic mouse system, we have developed a mouse model in which lung mesenchyme-specific deletion of TGF-ß receptor 2 gene (Tgfbr2) is achieved. Reduced airway branching accompanied by defective airway smooth muscle growth and later peripheral cystic lesions occurred when lung mesenchymal Tgfbr2 was deleted from embryonic day 13.5 to 15.5, resulting in postnatal death due to respiratory insufficiency. Although cell proliferation in both lung epithelium and mesenchyme was reduced, epithelial differentiation was not significantly affected. Tgfbr2 downstream Smad-independent ERK1/2 may mediate these mesenchymal effects of TGF-ß signaling through the GSK3ß-ß-catenin-Wnt canonical pathway in fetal mouse lung. Our study suggests that Tgfbr2-mediated TGF-ß signaling in prenatal lung mesenchyme is essential for lung development and maturation, and defective TGF-ß signaling in lung mesenchyme may be related to abnormal airway branching morphogenesis and congenital airway cystic lesions.


Assuntos
Cistos/metabolismo , Pneumopatias/patologia , Mesoderma/citologia , Receptores de Fatores de Crescimento Transformadores beta/metabolismo , Animais , Cistos/patologia , Células Epiteliais/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/genética , Pulmão/metabolismo , Pulmão/patologia , Pneumopatias/metabolismo , Camundongos , Camundongos Transgênicos , Morfogênese/efeitos dos fármacos , Morfogênese/fisiologia , Organogênese/fisiologia , Proteínas Serina-Treonina Quinases/metabolismo , Receptor do Fator de Crescimento Transformador beta Tipo II/metabolismo
9.
PLoS Negl Trop Dis ; 15(4): e0009370, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33909640

RESUMO

BACKGROUND: The diagnosis of cystic echinococcosis (CE) is primarily based on imaging, while serology should be applied when imaging is inconclusive. CE cyst stage has been reported among the most important factors influencing the outcome of serodiagnosis. We performed a systematic review and meta-analysis of the relation between cyst stage of hepatic CE and diagnostic sensitivity of serological tests, to evaluate whether their relation is a consistent finding and provide guidance for the interpretation of results of serological tests. METHODOLOGY/PRINCIPAL FINDINGS: MEDLINE, EMBASE, CENTRAL, and Lilacs databases were searched on December 1st 2019. Original studies published after 2003 (year of publication of the CE cyst classification), reporting sensitivity of serological tests applied to the diagnosis of human hepatic CE, as diagnosed and staged by imaging, were included. The quality of studies was assessed using the Newcastle-Ottawa Scale. Data from 14 studies were included in the meta-analysis. Summary estimates of sensitivities and 95% confidence intervals were obtained using random effects meta-analysis. Overall, test sensitivity was highest in the presence of CE2 and CE3 (CE3a and/or CE3b), and lowest in the presence of CE5 and CE4 cysts. ELISA, ICT and WB showed the highest sensitivities, while IHA performed worst. CONCLUSIONS/SIGNIFICANCE: The results of our study confirm the presence of a clear and consistent relation between cyst stage and serological tests results. Limitations of evidence included the heterogeneity of the antigenic preparations used, which prevented to determine whether the relation between cyst stage and sensitivity was influenced by the type of antigenic preparation, the paucity of studies testing the same panel of sera with different assays, and the lack of studies assessing the performance of the same assay in both field and hospital-based settings. Our results indicate the absolute need to consider cyst staging when evaluating serological results of patients with hepatic CE.


Assuntos
Cistos/patologia , Equinococose Hepática/diagnóstico , Echinococcus/imunologia , Testes Sorológicos/métodos , Animais , Anticorpos Anti-Helmínticos/sangue , Equinococose Hepática/sangue , Equinococose Hepática/parasitologia , Echinococcus/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Humanos , Sensibilidade e Especificidade , Fatores de Tempo
10.
Indian J Pathol Microbiol ; 64(2): 343-346, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33851631

RESUMO

Background: Renal oncocytomas are benign epithelial tumors usually detected incidentally. They are typically solid,well-circumscribed,homogenous,mahoganybrown with a central stellate scar.Sometimes,they can have cystic degenerationand rarely present as a multilocular cyst which can be mistaken for other cystic renal carcinomas. Methods: We describe a case of incidentally detected multilocular cystic renal oncocytoma having an unusual gross appearance of multiloculation with perinephric fat invasion. The tumor exhibited tubulocystic architecture posed a diagnostic dilemma. Detailed study of multiple sections coupled with immunohisto chemistry helped elucidate the diagnosis. Till date, only eight cases of multicystic renal oncocytoma have been reported in the English literature. Conclusions: We emphasize the importance of awareness of this unusual morphologic variation to ensure correct diagnosis.


Assuntos
Adenoma Oxífilo/diagnóstico , Adenoma Oxífilo/patologia , Cistos/patologia , Neoplasias Renais/diagnóstico , Neoplasias Renais/patologia , Rim/patologia , Idoso , Carcinoma de Células Renais/diagnóstico , Diagnóstico Diferencial , Humanos , Imuno-Histoquímica , Masculino
11.
World Neurosurg ; 150: e491-e499, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33744422

RESUMO

OBJECTIVE: Although the spontaneous regression of pseudotumors after posterior fusion has been reported, the predictive factors remain unclear. We examined the radiological features that predict for the regression of retro-odontoid pseudotumors after posterior fusion, with a specific focus on cyst formation. METHODS: We included 28 patients with a diagnosis of retro-odontoid pseudotumor using preoperative magnetic resonance imaging. The radiographic parameters and pseudotumor thickness were measured pre- and postoperatively. The regression rate for each pseudotumor was calculated. The presence of a cyst around the retro-odontoid pseudotumor was investigated. If present, the cyst thickness was measured. To elucidate the predictors for the postoperative regression of pseudotumors, the patients were divided into 2 cohorts: the regression group with a regression rate >40% and the no-regression group with a regression rate of <40%. Multivariate logistic regression analysis, including the demographic data and preoperative radiographic parameters as independent variables, was performed. RESULTS: The mean pseudotumor size had decreased significantly from 8.8 ± 3.6 mm preoperatively to 5.3 ± 2.0 mm postoperatively (P < 0.0001). The mean regression rate was 35.9% during a magnetic resonance imaging follow-up period of 8.6 months (range, 6-12 months). Cystic lesions were noted in 10 patients (35.7%) preoperatively. The mean cyst size was 4.7 ± 1.9 mm. All cysts were located dorsal to the pseudotumors and were involved at the maximum spinal compression levels. Nevertheless, all the cysts had disappeared postoperatively. Multivariate logistic regression analysis revealed that the pseudotumor regression group had had a significantly greater proportion of cysts (57.1% vs. 14.3%; odds ratio, 11.7; P = 0.013). CONCLUSIONS: The presence of cystic lesions protruding from retro-odontoid pseudotumors might serve as a predictive factor for the spontaneous regression of pseudotumors after posterior fusion.


Assuntos
Cistos/patologia , Doenças da Coluna Vertebral/cirurgia , Fusão Vertebral , Idoso , Articulação Atlantoaxial/patologia , Articulação Atlantoaxial/cirurgia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Processo Odontoide/patologia , Remissão Espontânea , Estudos Retrospectivos
12.
Afr J Paediatr Surg ; 18(2): 109-110, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33642411

RESUMO

Duplication cysts of gastrointestinal tract are rare congenital abnormalities found in 0.2% of children. We report a rare case of a ruptured duplication cyst of transverse colon in a 7-year-old female child who presented with abdominal pain and mass in the right iliac fossa. We assumed it as an appendicular mass; however, it turned out to be a ruptured duplication cyst of transverse colon. Only two cases of duplication cyst of transverse colon have been reported yet in the literature.


Assuntos
Colo Transverso/anormalidades , Cistos/congênito , Cistos/patologia , Apêndice , Criança , Cistos/cirurgia , Feminino , Humanos , Ruptura Espontânea
13.
J Vet Diagn Invest ; 33(3): 479-497, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33666111

RESUMO

Hair follicle neoplasms occur in many different species, including humans. In domestic animals, they are most common in dogs. Most hair follicle tumors are benign, but malignant neoplasms can also occur. To diagnose hair follicle neoplasms, a thorough knowledge of follicular anatomy is important, given that follicular tumors are classified according to the differentiation pattern seen in the corresponding part of the normal hair follicle. This review focuses on the key diagnostic features of hair follicle tumors and follicular cysts in dogs and cats.


Assuntos
Doenças do Gato/diagnóstico , Cistos/veterinária , Doenças do Cão/diagnóstico , Doenças do Cabelo/veterinária , Folículo Piloso/patologia , Neoplasias Cutâneas/veterinária , Animais , Doenças do Gato/patologia , Gatos , Cistos/diagnóstico , Cistos/patologia , Doenças do Cão/patologia , Cães , Doenças do Cabelo/diagnóstico , Doenças do Cabelo/patologia , Neoplasias Cutâneas/diagnóstico , Neoplasias Cutâneas/patologia
14.
Biochim Biophys Acta Gen Subj ; 1865(6): 129859, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33581251

RESUMO

BACKGROUND: Giardia lamblia differentiates into resistant cysts as an established model for dormancy. Myeloid leukemia factor (MLF) proteins are important regulators of cell differentiation. Giardia possesses a MLF homolog which was up-regulated during encystation and localized to unknown cytosolic vesicles named MLF vesicles (MLFVs). METHODS: We used double staining for visualization of potential factors with role in protein metabolism pathway and a strategy that employed a deletion mutant, CDK2m3, to test the protein degradation pathway. We also explored whether autophagy or proteasomal degradation are regulators of Giardia encystation by treatment with MG132, rapamycin, or chloroquine. RESULTS: Double staining of MLF and ISCU or CWP1 revealed no overlap between their vesicles. The aberrant CDK2m3 colocalized with MLFVs and formed complexes with MLF. MG132 increased the number of CDK2m3-localized vesicles and its protein level. We further found that MLF colocalized and interacted with a FYVE protein and an ATG8-like (ATG8L) protein, which were up-regulated during encystation and their expression induced Giardia encystation. The addition of MG132, rapamycin, or chloroquine, increased their levels and the number of their vesicles, and inhibited the cyst formation. MLF and FYVE were detected in exosomes released from culture. CONCLUSIONS: The MLFVs are not mitosomes or encystation-specific vesicles, but are related with degradative pathway for CDK2m3. MLF, FYVE, and ATG8L play a positive role in encystation and function in protein clearance pathway, which is important for encystation and coordinated with Exosomes. GENERAL SIGNIFICANCE: MLF, FYVE, and ATG8L may be involved an encystation-induced protein metabolism during Giardia differentiation.


Assuntos
Proteínas de Ciclo Celular/metabolismo , Quinase 2 Dependente de Ciclina/metabolismo , Cistos/patologia , Giardia lamblia/metabolismo , Encistamento de Parasitas , Proteínas de Protozoários/metabolismo , Proteínas de Ciclo Celular/genética , Quinase 2 Dependente de Ciclina/genética , Cistos/metabolismo , Giardia lamblia/genética , Giardia lamblia/crescimento & desenvolvimento , Proteínas de Protozoários/genética
15.
Proc Natl Acad Sci U S A ; 118(6)2021 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-33536341

RESUMO

Tuberous sclerosis complex (TSC) is caused by mutations in either TSC1 or TSC2 genes and affects multiple organs, including kidney, lung, and brain. In the kidney, TSC presents with the enlargement of benign tumors (angiomyolipomata) and cysts, which eventually leads to kidney failure. The factors promoting cyst formation and tumor growth in TSC are incompletely understood. Here, we report that mice with principal cell-specific inactivation of Tsc1 develop numerous cortical cysts, which are overwhelmingly composed of hyperproliferating A-intercalated (A-IC) cells. RNA sequencing and confirmatory expression studies demonstrated robust expression of Forkhead Transcription Factor 1 (Foxi1) and its downstream targets, apical H+-ATPase and cytoplasmic carbonic anhydrase 2 (CAII), in cyst epithelia in Tsc1 knockout (KO) mice but not in Pkd1 mutant mice. In addition, the electrogenic 2Cl-/H+ exchanger (CLC-5) is significantly up-regulated and shows remarkable colocalization with H+-ATPase on the apical membrane of cyst epithelia in Tsc1 KO mice. Deletion of Foxi1, which is vital to intercalated cells viability and H+-ATPase expression, completely abrogated the cyst burden in Tsc1 KO mice, as indicated by MRI images and histological analysis in kidneys of Foxi1/Tsc1 double-knockout (dKO) mice. Deletion of CAII, which is critical to H+-ATPase activation, caused significant reduction in cyst burden and increased life expectancy in CAII/Tsc1 dKO mice vs. Tsc1 KO mice. We propose that intercalated cells and their acid/base/electrolyte transport machinery (H+-ATPase/CAII/CLC-5) are critical to cystogenesis, and their inhibition or inactivation is associated with significant protection against cyst generation and/or enlargement in TSC.


Assuntos
Anidrase Carbônica II/genética , Fatores de Transcrição Forkhead/genética , Insuficiência Renal/genética , Proteína 1 do Complexo Esclerose Tuberosa/genética , Animais , Cistos/genética , Cistos/patologia , Modelos Animais de Doenças , Humanos , Rim/metabolismo , Rim/patologia , Camundongos , Mutação/genética , ATPases Translocadoras de Prótons/genética , Insuficiência Renal/patologia , Canais de Cátion TRPP/genética , Esclerose Tuberosa
16.
BMC Biol ; 19(1): 25, 2021 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-33557824

RESUMO

BACKGROUND: Biomarker discovery remains a major challenge for predictive medicine, in particular, in the context of chronic diseases. This is true for the widespread protozoan Toxoplasma gondii which establishes long-lasting parasitism in metazoans, humans included. This microbe successively unfolds distinct genetic programs that direct the transition from high to low replicative potential inside host cells. As a slow-replicating cell, the T. gondii bradyzoite developmental stage persists enclosed in a cyst compartment within tissues including the nervous system, being held by a sustained immune equilibrium which accounts for the prolonged clinically silent phase of parasitism. Serological surveys indicate that nearly one third of the human population has been exposed to T. gondii and possibly host bradyzoites. Because any disruption of the immune balance drives the reverse transition from bradyzoite to fast replicating tachyzoite and uncontrolled growth of the latter, these people are at risk for life-threatening disease. While serological tests for discriminating recent from past infection are available, there is yet no immunogenic biomarker used in the serological test to allow ascertaining the presence of persistent bradyzoites. RESULTS: Capitalizing on genetically engineered parasites induced to produce mature bradyzoites in vitro, we have identified the BCLA/MAG2 protein being restricted to the bradyzoite and the cyst envelope. Using laboratory mice as relevant T. gondii host models, we demonstrated that BCLA/MAG2 drives the generation of antibodies that recognize bradyzoite and the enveloping cyst structure. We have designed an ELISA assay based on a bacterially produced BCLA recombinant polypeptide, which was validated using a large collection of sera from mice of different genetic backgrounds and infected with bcla+ or bcla-null cystogenic and non-cystogenic T. gondii strains. To refine the design of the ELISA assay, we applied high-resolution BCLA epitope mapping and identified a specific combination of peptides and accordingly set up a selective and sensitive ELISA assay which allowed the detection of anti-BCLA/MAG2 antibodies in the sera of human patients with various forms of toxoplasmosis. CONCLUSIONS: We brought proof of principle that anti-BCLA/MAG2 antibodies serve as specific and sensitive serological markers in the perspective of a combinatorial strategy for detection of persistent T. gondii parasitism.


Assuntos
Encéfalo/parasitologia , Cistos/patologia , Toxoplasma/fisiologia , Toxoplasmose/diagnóstico , Biomarcadores/metabolismo , Doença Crônica , Cistos/diagnóstico , Cistos/parasitologia , Testes Sorológicos , Toxoplasmose/parasitologia , Toxoplasmose/patologia
17.
Medicine (Baltimore) ; 100(7): e24362, 2021 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-33607770

RESUMO

ABSTRACT: This study investigates the effect of 2 laparoscopic methods on ovarian reserve in patients of reproductive age with endometriomas.This was a retrospective study performed at a tertiary medical center from Jan 1st to Dec 31st, 2016. Laparoscopic cystectomy (group 1, 46 patients) and laparoscopic ovarian drainage and ablation with bipolar coagulation at low power (group 2, 30 patients) were performed to treat endometriomas larger than 3 cm. Anti-Müllerian hormone was used to assess ovarian reserve before and after surgery.There were no statistically significant differences in patients' baseline clinical characteristics, endometriotic stage, operative time, and follow-up time between the groups. The mean serum anti-Müllerian hormone concentration decreased significantly from 4.25 ng/ml to 3.40 ng/ml in group 1 compared with 4.47 ng/ml to 3.95 ng/ml in group 2 (P  = .04). Pregnancy rates were 71.05% in group 1 and 73.08% in group 2, with a mean follow-up of 30.40 months and 32.35 months (P  > .99), respectively. Although there was no statistical significance, the recurrence rate in group 1 was lower than that in group 2 (4.35% vs 16.67%, respectively; P = .11). The mean diameter of recurrent cysts was 1.75 cm in group 1 and 1.54 cm in group 2 (P = .13).Appropriate laparoscopic electrocautery of the endometrioma wall with a bipolar instrument may be a valid alternative to traditional laparoscopic cystectomy, with less effects on ovarian reserve.


Assuntos
Cistos/cirurgia , Técnicas de Ablação Endometrial/métodos , Endometriose/cirurgia , Laparoscopia/métodos , Reserva Ovariana , Adulto , Hormônio Antimülleriano/sangue , Cistos/patologia , Endometriose/patologia , Feminino , Humanos , Gravidez , Estudos Retrospectivos
18.
Am J Pathol ; 191(5): 902-920, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33549515

RESUMO

DNA damage and alterations in DNA damage response (DDR) signaling could be one of the molecular mechanisms mediating focal kidney cyst formation in autosomal dominant polycystic kidney disease (ADPKD). The aim of this study was to test the hypothesis that markers of DNA damage and DDR signaling are increased in human and experimental ADPKD. In the human ADPKD transcriptome, the number of up-regulated DDR-related genes was increased by 16.6-fold compared with that in normal kidney, and by 2.5-fold in cystic compared with that in minimally cystic tissue (P < 0.0001). In end-stage human ADPKD tissue, γ-H2A histone family member X (H2AX), phosphorylated ataxia telangiectasia and radiation-sensitive mutant 3 (Rad3)-related (pATR), and phosphorylated ataxia telangiectasia mutated (pATM) localized to cystic kidney epithelial cells. In vitro, pATR and pATM were also constitutively increased in human ADPKD tubular cells (WT 9-7 and 9-12) compared with control (HK-2). In addition, extrinsic oxidative DNA damage by hydrogen peroxide augmented γ-H2AX and cell survival in human ADPKD cells, and exacerbated cyst growth in the three-dimensional Madin-Darby canine kidney cyst model. In contrast, DDR-related gene expression was only transiently increased on postnatal day 0 in Pkd1RC/RC mice, and not altered at later time points up to 12 months of age. In conclusion, DDR signaling is dysregulated in human ADPKD and during the early phases of murine ADPKD. The constitutive expression of the DDR pathway in ADPKD may promote survival of PKD1-mutated cells and contribute to kidney cyst growth.


Assuntos
Dano ao DNA , Rim Policístico Autossômico Dominante/genética , Transdução de Sinais , Animais , Linhagem Celular , Cistos/patologia , Cães , Células Epiteliais/patologia , Feminino , Humanos , Peróxido de Hidrogênio/metabolismo , Rim/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mutação , Fosforilação , Rim Policístico Autossômico Dominante/patologia , Regulação para Cima
19.
Sci Rep ; 11(1): 1190, 2021 01 13.
Artigo em Inglês | MEDLINE | ID: mdl-33441940

RESUMO

To assess extraspinal findings (ESFs) prevalence in lumbar spine MRI, including clinically significant findings using a systematic approach, and to determine their reporting rate. Lumbar spine MRI scans were retrospectively reviewed over 18 months by two radiologists. Reading discrepancies were resolved by consensus. ESFs were classified according to the involved system, clinical diagnosis, and clinical significance. The reporting rate was estimated by referring to the original report. There were 1509 ESFs in 1322/4250 patients with a substantial agreement between the two radiologists (kappa = 0.8). Almost half (621/1322) were in the 45-60 age group. Females represented 56.6% (748/1322). 74.2% (1120/1509) of the ESFs involved the urinary system among which 79.6% (892/1120) were renal cysts. Clinically significant findings represented 8.7% (131/1509) among which hydronephrosis represented 23% (30/131). First time detected malignant lesions represented 4.6% (6/131). ESFs reporting rate was 47.3%. 58.8% of the clinically significant ESFs were not reported. ESFs prevalence was 31.1%. The Urinary system was the most commonly involved. Most ESFs were benign warranting no further workup. However, clinically significant ESF were not infrequently detected. More than half of the clinically significant findings were not reported. A systematic review of MRI images is highly recommended to improve patient's outcome.


Assuntos
Vértebras Lombares/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Cistos/patologia , Feminino , Humanos , Achados Incidentais , Lactente , Imageamento por Ressonância Magnética/métodos , Masculino , Pessoa de Meia-Idade , Prevalência , Estudos Retrospectivos , Adulto Jovem
20.
Vet Ophthalmol ; 24(2): 195-202, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33444470

RESUMO

OBJECTIVE: To describe and discuss ventromedial orbital lacrimal gland or duct cysts (dacryops) in dogs with extensive bone defects based on their symptoms, results of diagnostic imaging and histopathological examination, and therapy and discuss their potential origin based on the morphology. ANIMALS STUDIED: Four dogs of different breeds, age, and sex were presented with a unilateral round, slow growing, indolent, and non-tender process ventromedial to the nasal canthus of the eye. PROCEDURES: Transverse computed tomography showed a low-density, non-contrast-enhancing cystic process ventromedial to the globe with extensive defects in the lacrimal and maxillary bones in all cases. The cystic character of the structure was confirmed by the aspiration of the brownish fluid without cellular and microbiological contents. For treatment, the cystic fluid was aspirated, and the sclerosing agent polidocanol was injected in three cases. Cystorhinostomy (nasal marsupialisation) was performed in one case as the first choice and in another case following failure of sclerotherapy. Histopathological examination of the cyst walls was performed in two cases and confirmed the diagnosis of dacryops. RESULTS: Follow-up between 2 and 18 months showed no recurrence and very good to excellent cosmetic results. CONCLUSIONS: Aberrant lacrimal gland or duct tissue with secondary development of dacryops should be included in the differential diagnoses of ventromedial orbital cysts. Large cysts near the lacrimal drainage system with extensive bone defects in dogs should be treated by nasal marsupialization. Treatments such as evacuation of the cyst and inducing sclerosis (sclerotherapy) should be reserved for exceptional cases.


Assuntos
Cistos/veterinária , Doenças do Cão/patologia , Doenças do Aparelho Lacrimal/veterinária , Aparelho Lacrimal/patologia , Maxila/patologia , Animais , Cistos/patologia , Doenças do Cão/diagnóstico por imagem , Doenças do Cão/terapia , Cães , Feminino , Seguimentos , Aparelho Lacrimal/diagnóstico por imagem , Doenças do Aparelho Lacrimal/diagnóstico por imagem , Doenças do Aparelho Lacrimal/patologia , Doenças do Aparelho Lacrimal/terapia , Masculino , Maxila/diagnóstico por imagem , Tomografia Computadorizada por Raios X/veterinária
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