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There are multiple factors associated with increased morbidity and mortality in COVID-19 patients, and advanced age is one such independent prognostic factor. It is well established that the multiorgan failure and death in COVID-19 patients are due to the hyperactivation of the NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome and the ensuing cytokine storm. Colchicine, a well-known anti-inflammatory drug, has been shown to inhibit the NLRP3 inflammasome in micromolar concentrations potently. It has the unique property of accumulating in leukocytes, which is the primary cause of the abnormal activation of the NLRP3 inflammasome in COVID-19. It has been shown that achieving inhibitory concentrations of colchicine in leucocytes requires treatment with higher doses. Our recent studies showed that treatment with higher doses of colchicine in both outpatient and inpatient settings is safe and results in remarkable cure rates and significantly decreased mortality rates, even in the most severely affected patients with multiple comorbidities and risk factors. The main risk factor for severe COVID-19 is age, especially over 85 years. Here, we present a unique case of a 101-year-old male who underwent two major emergency abdominal surgeries and contracted COVID-19 while in the hospital. Laboratory tests showed increased values of markers for severe COVID-19, including CRP, D-dimer, and ferritin. Increased opacities bilaterally paracardially and moderate right-side pleural effusions were detected on the chest X-ray. We initiated our high-dose colchicine treatment regimen, resulting in the patient's complete recovery and discharge. We are convinced that the administration of high-dose colchicine to high-risk COVID-19 patients should be mandatory.
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PURPOSE: This preliminary study aimed to analyze and identify differentially expressed miRNAs in Bulgarian patients with non-functioning pituitary neuroendocrine tumors (NFPitNET). The relationship between deregulated miRNAs and tumor invasiveness, recurrence, and size was determined. METHODS: Twenty patients with NFPitNET were selected and fresh pituitary tumor tissues were collected. RNA containing miRNAs were isolated using miRNAeasy mini kit and analyzed by quantitative real-time polymerase chain reaction (PCR) using LNA miRNA Cancer-Focus PCR Panel (Qiagen). RESULTS: Three miRNAs (miR-210-3p, miR-149-3p, and miR-29b-3p) were deregulated in invasive compared to non-invasive NFPitNETs. Differential expression of four-miRNA signatures - miRNA-17, miR-19, miR-106a, and miR-20, correlated with patient recurrence. CONCLUSION: This prospective pilot study selected a unique miRNA expression profile, that correlates with invasiveness and recurrence in non-functioning pituitary neuroendocrine tumors. Moreover, some of the selected miRNAs are reported for the first time in patients with this disease, shedding light on the molecular mechanisms involved in pituitary pathogenesis. The identified miRNAs demonstrate potential as biomarkers, deserving further investigation in a larger cohort to validate their clinical applicability.
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MicroRNAs , Invasividade Neoplásica , Recidiva Local de Neoplasia , Tumores Neuroendócrinos , Neoplasias Hipofisárias , Humanos , Neoplasias Hipofisárias/genética , Neoplasias Hipofisárias/patologia , Neoplasias Hipofisárias/metabolismo , Masculino , Feminino , MicroRNAs/genética , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Tumores Neuroendócrinos/genética , Tumores Neuroendócrinos/patologia , Tumores Neuroendócrinos/metabolismo , Recidiva Local de Neoplasia/genética , Adulto , Idoso , Projetos Piloto , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Regulação Neoplásica da Expressão GênicaRESUMO
COVID19 patients with severe infection have been observed to have elevated autoantibodies (AAs) against angiotensin II receptor type 1 (AT1R) and endothelin (ET) 1 receptor type A (ETAR), compared with healthy controls and patients with favorable (mild) infection. AT1R and ETAR are G proteincoupled receptors, located on vascular smooth muscle cells, fibroblasts, immune and endothelial cells, and are activated by angiotensin II (Ang II) and ET1 respectively. AAs that are specific for these receptors have a functional role similar to the natural ligands, but with a more prolonged vasoconstrictive effect. They also induce the production of fibroblast collagen, the release of reactive oxygen species and the secretion of proinflammatory cytokines (including IL6, IL8 and TNFα) by immune cells. Despite the presence of AAs in severe COVID19 infected patients, their contribution and implication in the severity of the disease is still not well understood and further studies are warranted. The present review described the major vascular homeostasis systems [ET and reninangiotensinaldosterone system (RAAS)], the vital regulative role of nitric oxide, the AAs, and finally the administration of angiotensin II receptor blockers (ARBs), so as to provide more insight into the interplay that exists among these components and their contribution to the severity, prognosis and possible treatment of COVID19.
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COVID-19 , Doenças Vasculares , Angiotensina II , Antagonistas de Receptores de Angiotensina , Inibidores da Enzima Conversora de Angiotensina , Colágeno , Células Endoteliais , Endotelinas , Humanos , Interleucina-6 , Interleucina-8 , Óxido Nítrico , Espécies Reativas de Oxigênio , Receptor Tipo 1 de Angiotensina , Receptor de Endotelina A , Receptores de Angiotensina , Fator de Necrose Tumoral alfaRESUMO
The current study investigated the expression signatures of miRNAs in lung adenocarcinoma (LUAD) and squamous cell lung carcinoma (LUSC). miRNA profiling was performed using microarray in 12 LUAD and 12 LUSC samples and adjacent normal tissues. In LUAD, 107 miRNAs were significantly deregulated, whereas 235 miRNAs were deregulated in LUSC. Twenty-six miRNAs were common between the 2 cancer subtypes and 8 were prioritized for validation, in addition to 6 subtype-specific miRNAs. The RT-qPCR validation samples included 50 LUAD, 50 LUSC, and adjacent normal tissues. Eight miRNAs were validated in LUAD: 3 upregulated - miR-7-5p, miR-375-5p, miR-6785-3p, and 5 downregulated - miR-101-3p, miR-139-5p, miR-140-3p, miR-144-3p, miR-195-5p. Ten miRNAs were validated in the LUSC group: 3 upregulated - miR-7-5p, miR-21-3p, miR-650, and 7 downregulated - miR-95-5p, miR-140-3p, miR-144-3p, miR-195-5p, miR-375, miR-744-3p, and miR-4689-3p. Reactome pathway analysis revealed that the target genes of the deregulated miRNAs in LUAD were significantly enriched in cell cycle, membrane trafficking, gene expression processes, and EGFR signaling, while in LUSC, they were enriched in the immune system, transcriptional regulation by TP53, and FGFR signaling. This study identified distinct miRNA profiles in LUSC and LUAD, which are common and specific miRNAs that could be further investigated as biomarkers for diagnosis and prognosis.
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Adenocarcinoma de Pulmão , Adenocarcinoma , Carcinoma Pulmonar de Células não Pequenas , Carcinoma de Células Escamosas , Neoplasias Pulmonares , MicroRNAs , Adenocarcinoma/genética , Adenocarcinoma de Pulmão/genética , Adenocarcinoma de Pulmão/patologia , Carcinoma Pulmonar de Células não Pequenas/patologia , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Células Epiteliais/patologia , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Pulmão/patologia , Neoplasias Pulmonares/patologia , MicroRNAs/genética , MicroRNAs/metabolismoRESUMO
At present, brain tumours remain one of the "hard-to-treat" malignancies with minimal improvement in patients' survival. Recently, miRNAs have been shown to correlate with oncogenesis and metastasis and have been investigated as potential biomarkers for diagnosis, prognosis and therapy prediction in different brain malignancies. The aim of the current study was to select an accurate and affordable brain tumour detection and grading approach. In the present study, we analysed the applicability of a restricted miRNA signature that could differentiate among patients with primary as well as metastatic brain tumours. Fresh tumour tissues were collected from Bulgarian patients (n = 38), including high-grade gliomas (n = 23), low-grade gliomas (n = 10) and brain metastases (n = 5) from lung cancer. Total RNAs enriched with microRNAs were isolated and differentially expressed miRNAs were analyzed by RT-qPCR using TaqMan Advanced miRNA assay. We selected a signature of miR-21, miR-10b, miR-7, miR-491 that showed good diagnostic potential in high-grade gliomas, low-grade gliomas and brain metastases compared with normal brain tissues. Our results showed that miR-10b could reliably differentiate brain metastases from high-grade gliomas, while miR-491 could distinguish low-grade from high-grade gliomas and brain metastases from low-grade gliomas. We observed that miR-21 and miR-7 correlated with disease recurrence, survival status and the Karnofsky Performance Status. The selected signature of miR-7, miR-21, miR-10b and miR-491 could be used as a highly accurate diagnostic, grading and prognostic biomarker in differentiating various types of brain tumours. Our data suggest that the 4-miRNAs signature could be further analysed for predicting treatment response and for future miRs-based targeted therapy. The ongoing studies on miRs-based targeted therapy related to our selected miRNA signature are also reviewed.
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Neoplasias Encefálicas , MicroRNAs , Biomarcadores Tumorais/genética , Encéfalo , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/terapia , Regulação Neoplásica da Expressão Gênica , Humanos , MicroRNAs/genética , Gradação de Tumores , PrognósticoRESUMO
INTRODUCTION: Recently, miRNAs have become popular molecules used as non-invasive biomarkers in cancer diseases. AIM: The aim of the study was to explore the expression of four miRNAs isoforms: miR-31-3p, miR-196a-5p, miR-210-3p and miR-424-5p in plasma and tissue samples from patients with advanced laryngeal squamous cell carcinoma (LSCC) and healthy controls. MATERIALS AND METHODS: Fresh-frozen tumour and normal laryngeal tissue as well as plasma samples were obtained from 22 patients diagnosed with advanced LSCC. The control group included plasma samples from 21 cancer-free volunteers. Total RNA (including miRNAs) extraction, reverse transcription and real time qPCR were the laboratory techniques used in the study. The obtained results were analyzed using SPSS software v. 23. RESULTS: We found that miR-31-3p, miR-196a-5p, and miR-210-3p levels were significantly elevated in laryngeal tumour tissue, but only the levels of miR-31-3p and miR-196a-5p were significantly upregulated in the plasma LSCC target group. Positive correlation was obtained for miR-31-3p (rs=0.443, p=0.039) and miR-196a-5p (rs=0.548; p=0.008) between plasma and adjacent tumour tissue LSCC samples. ROC analyses were used to evaluate the discriminative power of both miRNAs alone and in combination. The combination of miR-31-3p and miR-196a-5p showed best results with AUC=0.978 (95% CI: 0.945-1.000, p.
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Neoplasias Laríngeas , MicroRNAs/provisão & distribuição , Biomarcadores Tumorais/genética , Humanos , Neoplasias Laríngeas/genética , MicroRNAs/genética , Projetos Piloto , Carcinoma de Células Escamosas de Cabeça e PescoçoRESUMO
The hereditary transthyretin amyloidosis (ATTRv amyloidosis) is an autosomal dominant genetic disease characterized by amyloid formation in different tissues due to pathogenic variants in the TTR gene. Great heterogeneity in the penetrance and manifestation of ATTRv amyloidosis is observed. In Bulgaria, the most common TTR pathogenic variant is Glu89Gln. Other TTR pathogenic variants are also found - Val30Met, Ser77Phe, Gly47Glu and Ser52Pro. There is a proven founder effect for the Glu89Gln variant, thus the aim of the present study is to investigate the founder effect for the other TTR pathogenic variants in Bulgaria. Haplotype analysis was performed by using microsatellite markers close to the TTR gene. DNA samples from ATTRv amyloidosis patients and their healthy relatives were analyzed. Theoretical haplotype reconstruction was done with Arlequin v.3.01 software. The age of the most recent common ancestor (hypothetical founder) for the studied variants was calculated with the DMLE 2.2 software. In addition, DBS screening among 100 Roma newborns was done for the Gly47Glu TTR variant via direct Sanger sequencing. The reconstructed haplotypes of the patients were compared to their healthy relatives and to a control group of 40 healthy individuals. The results showed a possible founder effect for each of the studied variants. The Val30Met haplotype was compared to published haplotype data for this variant and no similarity was found. The result from the DBS screening showed no pathogenic TTR variants in exon 2 of the gene, so we considered the presence of the Gly47Glu variant in our population a sporadic event. With this study, we succeeded to gain a more complete picture of the population genetics of ATTRv amyloidosis in Bulgaria and made another step towards a more detailed understanding of the disease epidemiology.
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Neuropatias Amiloides Familiares , Pré-Albumina , Neuropatias Amiloides Familiares/genética , Bulgária , Efeito Fundador , Humanos , Recém-Nascido , Pré-Albumina/genéticaRESUMO
Laryngeal squamous cell carcinoma is a common malignancy in men. Bulgaria is one of the countries in Europe with the highest incidence and mortality rates of the aggressive, severe disease of laryngeal cancer. Proven etiological factors are the abuse of tobacco and alcohol beverages. Despite the progress of technologies of multimodal medical treatment, survival rates have not reached satisfactory levels. Over the last few decades, scientific and clinical research data have led to a growing interest in exploring potential biomarkers. In the last years, non-coding RNAs have become promising biomarkers. They are important key regulators in both normal and tumour specific biological processes as well as in the response to environmental factors and treatment, including chemo- and radiotherapy. Studies have shown ectopic expression of a number of ncRNAs in laryngeal cancer. Published data provide evidence of the lncRNAs and miRNAs that could help us better understand complex carcinogenesis in laryngeal cancer and would provide reliable diagnostic, prognostic and predictive biomarkers.
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Regulação Neoplásica da Expressão Gênica , Neoplasias Laríngeas/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Bulgária/epidemiologia , Humanos , Incidência , Neoplasias Laríngeas/epidemiologia , Neoplasias Laríngeas/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/epidemiologia , Carcinoma de Células Escamosas de Cabeça e Pescoço/metabolismoRESUMO
PURPOSE: Laryngeal cancer is one of most common and aggressive head and neck cancers with poor prognosis and great necessity for improvement of treatment modalities. MicroRNAs (miRs) are among the most investigated molecules recently due to their potential as diagnostic and prognostic biomarkers in cancer. The purpose of our study was to explore the association of certain clinicopathological features with the expression levels of some known cancer associated non-coding (nc) RNAs: miR-21 and miR-31 in both of their isoforms, miR-145-5p, miR-55-5p, miR-196a-5p, miR-210-3p, miR-221-3p, miR-222-3p, miR-424-5p, lncRNA MALAT1 and lncRNA HOTAIR. METHODS: Expression levels of the chosen markers were investigated in laryngeal squamous cell carcinoma (LSCC) and normal samples in 82 Bulgarian patients via RT-qPCR, and the results were analyzed with SPSS v23.0 statistical software. RESULTS: All of the explored ncRNAs were significantly deregulated in LSCC samples, suggesting their involvement in laryngeal carcinogenesis. New significant association were found between the expression levels of miR-21-5p, miR-222-3p, HOTAIR and family history. Moreover, miR-424-5p showed potential as marker for subglottic LSCC location, and "passenger" miR-31-3p was significantly upregulated in well and moderately differentiated LSCC. CONCLUSION: Our results enrich the knowledge about ncRNA involvement in LSCC tumorigenesis. Further studies are needed to evaluate the clinical utility of the differently expressed ncRNAs as potential diagnostic and prognostic biomarkers in LSCC.
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Carcinoma de Células Escamosas/genética , Neoplasias Laríngeas/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Bulgária , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Feminino , Humanos , Neoplasias Laríngeas/patologia , Masculino , Pessoa de Meia-IdadeRESUMO
Hereditary transthyretin amyloidosis is an autosomal dominant genetic disorder caused by missense mutations in the TTR gene resulting in amyloid formation of the transthyretin protein. Depending on the system affection, the manifestations may be different and high heterogeneity in the penetrance is observed. An endemic region in Bulgaria exists where the TTR mutation Glu89Gln is found with high frequency. This is a rare mutation and was probably introduced in the population by a common ancestor. This phenomenon, called "founder effect" was proved in carrier families by haplotype analysis of microsatellite markers showing linkage disequilibrium. Allele frequencies were analyzed and haplotype reconstruction was done with Arlequin v.3.01 software. The common ancestry of the carriers was demonstrated using additional data for their genealogies and microsatellite data from a control group of non-affected individuals. The results show that the mutation Glu89Gln is linked to one haplotype, called "hypothetical founder haplotype" which was compared to published haplotype data from other European patients and no similarity was found. Further population genetics studies of carriers of the Glu89Gln mutation from other endemic regions are required in order to clarify the geographical distribution of the mutation.
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Neuropatias Amiloides Familiares/genética , Efeito Fundador , Ácido Glutâmico/genética , Glutamina/genética , Pré-Albumina/genética , Adulto , Idoso , Neuropatias Amiloides Familiares/epidemiologia , Bulgária/epidemiologia , Feminino , Haplótipos , Humanos , Masculino , Repetições de Microssatélites/genética , Pessoa de Meia-Idade , MutaçãoRESUMO
TTR-related amyloidosis (ATTR) is manifested in two allelic forms: familial amyloid polyneuropathy (TTR-FAP) and cardiomyopathy (TTR-FAC), both caused by mutations in the TTR gene. The most prevalent mutation in Bulgaria is p.Glu89Gln. Markedly different age at onset and disease penetrance is noticed in Bulgarian p.Glu89Gln cases even in a single family or between genetically identical twins. The present study aimed to evaluate the transcription profile of the TTR gene in order to better understand the difference in disease onset and penetrance. Six p.Glu89Gln positive families were selected from our registry, based on intrafamilial differences in disease onset and penetrance. Plasma and urine specimens were collected from 13 patients and subjected to transcription analysis. Both mutant and wild type transcripts were visualized in a mixed transcription profile, which is the traditional model of autosomal gene expression. The results from a relative quantification of the mutant versus wild type transcript showed presence of the mutant transcript between 0.14 and 1.14 times against the wild type. In addition, monoallelic expression signature was also detected. Based on our results we propose a model of natural selection, which includes age-related allele exclusion or suppression: predominant expression of a wild type (at an early age) and mutant (at the process of ageing) alleles. The intrafamilial differences in disease onset and penetrance need to be considered in genetic counselling and in follow-up of mutation carriers.
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Substituição de Aminoácidos , Neuropatias Amiloides Familiares/genética , Cardiomiopatias/genética , Penetrância , Pré-Albumina/genética , Adulto , Idade de Início , Alelos , Bulgária , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Linhagem , Seleção GenéticaRESUMO
FGF8 specifies early tooth development by directing the migration of the early tooth founder cells to the site of tooth emergence. To date the effect of the FGF8 in adult dental pulp has not been studied. We have assessed the regenerative potential of FGF8 by evaluating changes in the proteome landscape of dental pulp following short- and long-term exposure to recombinant FGF8 protein. In addition, we carried out qRT PCR analysis to determine extracellular/adhesion gene marker expression and assessed cell proliferation and mineralization in response to FGF8 treatment. 2D and mass spectrometry data showed differential expression of proteins implicated in cytoskeleton/ECM remodeling and migration, cell proliferation and odontogenic differentiation as evidenced by the upregulation of gelsolin, moesin, LMNA, WDR1, PLOD2, COPS5 and downregulation of P4HB. qRT PCR showed downregulation of proteins involved in cell-matrix adhesion such as ADAMTS8, LAMB3 and ANOS1 and increased expression of the angiogenesis marker PECAM1. We have observed that, FGF8 treatment was able to boost dental pulp cell proliferation and to enhance dental pulp mineralization. Collectively, our data suggest that, FGF8 treatment could promote endogenous healing of the dental pulp via recruitment of dental pulp progenitors as well as by promoting their angiogenic and odontogenic differentiation. SIGNIFICANCE: Dental pulp cells (DP) have been studied extensively for the purposes of mineralized tissue repair, particularly for the reconstruction of hard and soft tissue maxillofacial defects. Canonical FGF signaling has been implicated throughout multiple stages of tooth development by regulating cell proliferation, differentiation, survival as well as cellular migration. FGF8 expression is indispensible for normal tooth development and particularly for the migration of early tooth progenitors to the sites of tooth emergence. The present study provides proteome and qRT PCR data with regard to the future application and biological relevance of FGF8 in dental regenerative medicine. AUTHORS WITH ORCID: Rozaliya Tsikandelova - 0000-0003-0178-3767 Zornitsa Mihaylova - 0000-0003-1748-4489 Sébastien Planchon - 0000-0002-0455-0574 Nikolay Ishkitiev - 0000-0002-4351-5579.
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Polpa Dentária/citologia , Fator 8 de Crescimento de Fibroblasto/farmacologia , Proteoma/metabolismo , Regeneração/efeitos dos fármacos , Adulto , Diferenciação Celular , Movimento Celular , Proliferação de Células/efeitos dos fármacos , Polpa Dentária/efeitos dos fármacos , Polpa Dentária/fisiologia , Regulação da Expressão Gênica , Humanos , Minerais/metabolismo , Reação em Cadeia da Polimerase , Proteoma/efeitos dos fármacos , Proteoma/fisiologiaRESUMO
Von Hippel-Lindau syndrome is an autosomal-dominant disease characterized by the formation of various tumours and cysts in many different parts of the body. Von Hippel-Lindau syndrome is caused by VHL gene mutations leading to production of impaired tumor suppressor Von Hippel-Lindau syndrome protein or its complete absence. PURPOSE: To study five patients with clinically suspected Von Hippel-Lindau syndrome, who were referred for molecular genetic testing. METHODS: Sanger sequencing of the coding regions of the VHL gene. RESULTS: Five clinically relevant germline mutations were detected. One of the pathogenic variants has not been previously reported. This novel mutation is a complex mutation event combining a duplication and an indel, rearranging exon 3 of the VHL gene - c. [516_517dupGTCAAGCCT; 532_542delCTGGACATCGTinsATTA], p. (Glu173Serfs*4). CONCLUSION: Overall, our results showed that the diagnosis of Von Hippel-Lindau syndrome in our country is difficult most probably because of its heterogeneous clinical manifestation and insufficient knowledge on the diagnostic criteria for the disease. From genetic point of view our results add some novel data on the mutation profile of the VHL gene. In order to prove or revise the diagnosis, early genetic testing is strongly recommended in affected patients and their family members to ensure appropriate follow-up and treatment of the malignancies.
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Proteína Supressora de Tumor Von Hippel-Lindau/genética , Doença de von Hippel-Lindau/diagnóstico , Adolescente , Adulto , Bulgária , Análise Mutacional de DNA , Feminino , Testes Genéticos , Humanos , Pessoa de Meia-Idade , Biologia Molecular , Mutação , Doença de von Hippel-Lindau/genéticaRESUMO
OBJECTIVE: Platelet-derived growth factor-BB (PDGF-BB) is one of the most abundant growth factors in platelet derived products and has been shown to stimulate regeneration after tissue injury. There is a population of mesenchymal stem cells (MSC) in human periodontal ligament (PDL) which can contribute to tissue regeneration under appropriate conditions. DESIGN: PDL cells were isolated and characterized using stem cell and differentiation markers via immunofluorescence and flow cytometry and then cultured in vitro and treated with different concentrations of PDGF-BB. The effect of PDGF-BB on cell proliferation, stem cell and differentiation markers expression, soluble collagen production, lysyl oxidase (LOX) activity, alkaline phosphatase (ALP) activity and calcium nodules formation was assessed. RESULTS: PDGF-BB stimulated the proliferation of cells with the maximum effect at 50ng/mL. The growth factor increased the expression of stem cell markers and SPARC; Col1a2 expression was decreased, whereas the expression of Col3a1 remain unchanged. Soluble collagen production, ALP activity and calcium nodules formation were also significantly decreased by PDGF-BB; LOX activity was significantly increased. CONCLUSIONS: PDGF-BB is a powerful promoter of cell proliferation and increases the expression of stem cell markers; inhibites collagen production and mineraliration but accelerates the maturation of collagen chains through increased LOX activity and SPARC expression.
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Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Ligamento Periodontal/citologia , Fator de Crescimento Derivado de Plaquetas/farmacologia , Adolescente , Adulto , Biomarcadores/análise , Células Cultivadas , Feminino , Citometria de Fluxo , Imunofluorescência , Humanos , Técnicas In Vitro , Masculino , Dente Serotino/cirurgiaRESUMO
OBJECTIVE: To describe and provide a comprehensive overview on the development, use and efficacy of autologous platelet concentrates in different in vitro and in vivo studies focusing on oral and maxillofacial pathologies. MATERIALS AND METHODS: Present work employs an extensive critical overview of the literature on the development and application of platelet concentrates. RESULTS: Platelet concentrates are innovative endogenous therapeutic agents which gained a lot of interest in different medical and dental disciplines due to their potential ability to stimulate and increase regeneration of soft and hard tissues. The effect of platelet-derived products is considered to be a result of the high number of platelets which contain a wide range of growth factors. They are not just therapeutic products but autologous blood concentrates containing active molecules. The quality of platelet concentrates may vary according to the individual physical state of donors making it difficult to to compare the outcomes of their application. Although, there are many studies analyzing the properties of these biomaterials both in vivo and in vitro, a consensus regarding their efficacy still has to be reached. CONCLUSION: Evidences described in the literature on the efficacy of platelet concentrates in procedures in oral and maxillofacial region are controversial and limited. In order to clarify the real advantages and priorities for the patients, when the blood-derived products are applied, further in vitro and in vivo research about the activity of PRP and PRF on the dental cells biology should be conducted.
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Plaquetas/fisiologia , Procedimentos Cirúrgicos Bucais/métodos , Fibrina/uso terapêutico , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/uso terapêutico , Plasma Rico em Plaquetas/fisiologia , Regeneração/fisiologia , Células-Tronco/fisiologia , Engenharia Tecidual/métodosRESUMO
Sézary syndrome is a primary cutaneous T-cell lymphoma characterized by pruritic erythroderma, peripheral lymphadenopathy and the presence of malignant T cells in the blood. Unequivocal detection of malignant cells in patients with Sézary syndrome is of important diagnostic, prognostic and therapeutic value. However, no single Sézary syndrome specific cell surface marker has been identified. In a cohort of patients with Sézary syndrome, CD164 expression on total CD4+ lymphocytes was significantly upregulated compared with healthy controls. CD164 expression was in most cases limited to CD4+CD26- malignant T lymphocytes, unequivocally identified using flow-cytometry by the expression of a specific Vß clone for each patient. Increased expression of CD164 may be a promising diagnostic parameter and a potential target for a CD164-linked therapeutic approach in Sézary syndrome.
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Biomarcadores Tumorais/sangue , Linfócitos T CD4-Positivos/imunologia , Síndrome de Sézary/sangue , Neoplasias Cutâneas/sangue , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Células Cultivadas , Dipeptidil Peptidase 4/sangue , Endolina/sangue , Feminino , Citometria de Fluxo , Humanos , Imunofenotipagem/métodos , Masculino , Pessoa de Meia-Idade , Fenótipo , Síndrome de Sézary/diagnóstico , Síndrome de Sézary/imunologia , Síndrome de Sézary/terapia , Neoplasias Cutâneas/diagnóstico , Neoplasias Cutâneas/imunologia , Neoplasias Cutâneas/terapia , Resultado do TratamentoRESUMO
PURPOSE: Metastatic prostate cancer (PCa) is one of the leading causes of death in men worldwide. We report Bulgarian patients with strongly aggressive, castration-resistant PCa. METHODS: PCA3 overexpression, GSTP1 promoter hyper-methylation, TMPRSS2-ERG gene fusions, IVS1-27G>A in the KLF6 gene and mutations in androgen receptor (AR) gene, for diagnostic purposes were assessed. PCR, real-time PCR (RT-PCR), sequencing, and bisulfite conversion of DNA were applied. We correlated the molecular data to the histological and clinical findings. RESULTS: The obtained molecular profile in 11 PCa Bulgarian patients coincided with the clinico-histological data of strongly aggressive PCa. Association was detected between the tumor stage (assessed by TNM as T3 and T4) and the detected molecular profile of aggressive cancer behavior with one exception, assessed as T2. None of our patients had positive family history of prostate cancer and no somatic mutations were detected in the AR gene. All patients showed normal genotype with respect to the KLF6 IVS1- 27G>A polymorphism. The rest of the markers were positive in fresh prostatic tissues and biopsies from all patients, whereas only one blood sample showed triple positive result. CONCLUSIONS: The appearance of PCa-specific markers in blood was considered as a predictor for a PCa (micro) dissemination into the circulation. The GSTP1 promoter hypermethylation is the earliest epigenetic alteration, which indicates cancerous changes and the first and long-lasting marker that is detectable in blood circulation. The molecular profile needs to be strictly monitored during treatment, which is of great help in determining the patient's individual response to therapy.
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Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Idoso , Bulgária , Metilação de DNA , Glutationa S-Transferase pi/genética , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Proteínas de Fusão Oncogênica/genética , Regiões Promotoras GenéticasRESUMO
To correlate the molecular data to the clinicopathological parameters in Bulgarian prostate cancer patients. PCA3 overexpression, TMPRSS2-ERG gene fusion, GSTP1 promoter hypermethylation, somatic mutations in the AR gene and the IVS1-27G > A polymorphism in the KLF6 gene were studied. A total of 148 patients were analyzed: 16 aggressive PCa, 83 non-aggressive PCa, 25 BPH and 24 chronic inflammatory diseases. Real-time RT-PCR, DNA sequencing, and bisulfite conversion of DNA, were applied. All cases with aggressive PCa before treatment were tested positive for PCA3 overexpression, expression of a T2-ERG gene fusion product and GSTP1 promoter hypermethylation. No somatic mutations were detected in the AR gene and all patients showed normal KLF6-IVS1-27G > A genotype. The TMPRSS2-ERG positive status correlates with moderate to poorly differentiated prostate tumors and it is considered as unfavorable disease predictor. Positive GSTP1 promoter hypermethylation seems to be highly specific and the earliest epigenetic change in the prostate gland, which indicates the beginning of the pathological process. The appearance of positive molecular markers in blood was considered as a predictor of PCa dissemination. GSTP1 promoter hypermethylation was found as the earliest and a long-lasting epigenetic marker in blood samples of PCa patients, which makes it suitable as a marker for treatment follow-up. The molecular profile of prostate cancer needs to be strictly monitored during the course of disease treatment, which is of a great help in determining the patient's individual therapy response.
Assuntos
Biomarcadores Tumorais/genética , Glutationa S-Transferase pi/genética , Fatores de Transcrição Kruppel-Like/genética , Hiperplasia Prostática/patologia , Neoplasias da Próstata/patologia , Proteínas Proto-Oncogênicas/genética , Receptores Androgênicos/genética , Serina Endopeptidases/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Bulgária/epidemiologia , Metilação de DNA , Seguimentos , Regulação Neoplásica da Expressão Gênica , Humanos , Fator 6 Semelhante a Kruppel , Masculino , Pessoa de Meia-Idade , Mutação/genética , Gradação de Tumores , Estadiamento de Neoplasias , Polimorfismo Genético/genética , Prognóstico , Regiões Promotoras Genéticas/genética , Hiperplasia Prostática/epidemiologia , Hiperplasia Prostática/genética , Neoplasias da Próstata/epidemiologia , Neoplasias da Próstata/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The heavy chain 1 of cytoplasmic dynein (DYNC1H1) is responsible for movement of the motor complex along microtubules and recruitment of dynein components. Mutations in DYNC1H1 are associated with spinal muscular atrophy (SMA), hereditary motor and sensory neuropathy (HMSN), cortical malformations, or a combination of these. Combining linkage analysis and whole-exome sequencing, we identified a novel dominant defect in the DYNC1H1 tail domain (c.1792C>T, p.Arg598Cys) causing axonal HMSN. Mutation analysis of the tail region in 355 patients identified a de novo mutation (c.791G>T, p.Arg264Leu) in an isolated SMA patient. Her phenotype was more severe than previously described, characterized by multiple congenital contractures and delayed motor milestones, without brain malformations. The mutations in DYNC1H1 increase the interaction with its adaptor BICD2. This relates to previous studies on BICD2 mutations causing a highly similar phenotype. Our findings broaden the genetic heterogeneity and refine the clinical spectrum of DYNC1H1, and have implications for molecular diagnostics of motor neuron diseases.
Assuntos
Doença de Charcot-Marie-Tooth/genética , Dineínas do Citoplasma/genética , Dineínas/metabolismo , Atrofia Muscular Espinal/genética , Mutação , Doença de Charcot-Marie-Tooth/fisiopatologia , Dineínas do Citoplasma/metabolismo , Humanos , Proteínas Associadas aos Microtúbulos/metabolismo , Atrofia Muscular Espinal/fisiopatologia , Estrutura Terciária de Proteína/genéticaRESUMO
The current definitive treatment for acute or chronic liver condition, that is, cirrhosis, is liver transplantation from a limited number of donors, which might cause complications after donation. Hence, bone marrow stem cell transplantation has been developed, but the risk of carcinogenesis remains. We have recently developed a protocol for hepatic differentiation of CD117(+) stem cells from human exfoliated deciduous teeth (SHED). In the present study, we examine whether SHED hepatically differentiated (hd) in vitro could be used to treat acute liver injury (ALI) and secondary biliary cirrhosis. The CD117(+) cell fraction was magnetically separated from SHED and then differentiated into hepatocyte-like cells in vitro. The cells were transplanted into rats with either ALI or induced secondary biliary cirrhosis. Engraftment of human liver cells was determined immunohistochemically and by in situ hybridization. Recovery of liver function was examined by means of histochemical and serological tests. Livers of transplanted animals were strongly positive for human immunohistochemical factors, and in situ hybridization confirmed engraftment of human hepatocytes. The tests for recovery of liver function confirmed the presence of human hepatic markers in the animals' blood serum and lack of fibrosis and functional integration of transplanted human cells into livers. No evidence of malignancy was found. We show that in vitro hdSHED engraft morphologically and functionally into the livers of rats having acute injury or secondary biliary cirrhosis. SHED are readily accessible adult stem cells, capable of proliferating in large numbers before differentiating in vitro. This makes SHED an appropriate and safe stem cell source for regenerative medicine.