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Ovarian cancer (OC) is usually diagnosed late due to its nonspecific symptoms and lack of reliable tools for early diagnostics and screening. OC studies concentrate on the search for new biomarkers and therapeutic targets. This study aimed to validate the MFAP5 gene, and its encoded protein, as a potential prognostic biomarker. In our previous study, we found that patients with high-grade serous OC who had higher MFAP5 mRNA levels had shorter survival, as compared with those with lower levels. Here, we used the Kaplan-Meier Plotter and CSIOVDB online tools to analyze possible associations of MFAP5 expression with survival and other clinico-pathological features. In these analyses, higher MFAP5 mRNA expression was observed in the more advanced FIGO stages and high-grade tumors, and was significantly associated with shorter overall and progression-free survival. Next, we analyzed the expression of the MFAP5 protein by immunohistochemistry (IHC) in 108 OC samples and tissue arrays. Stronger MFAP5 expression was associated with stronger desmoplastic reaction and serous vs. non-serous histology. We found no significant correlation between IHC results and survival, although there was a trend toward shorter survival in patients with the highest IHC scores. We searched for co-expressed genes/proteins using cBioPortal and analyzed potential MFAP5 interaction networks with the STRING tool. MFAP5 was shown to interact with many extracellular matrix proteins, and was connected to the Notch signaling pathway. Therefore, although not suitable as a prognostic biomarker for evaluation with a simple diagnostic tool like IHC, MFAP5 is worth further studies as a possible therapeutic target.
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Proteínas Contráteis , Neoplasias Ovarianas , Humanos , Feminino , Proteínas Contráteis/genética , Microfibrilas/metabolismo , Prognóstico , Peptídeos e Proteínas de Sinalização Intercelular , Neoplasias Ovarianas/diagnóstico , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , RNA Mensageiro/metabolismo , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismoRESUMO
High-grade serous ovarian carcinoma (HGSOC) is the most frequent histological type of ovarian cancer and the one with worst prognosis. Unfortunately, the majority of established ovarian cancer cell lines which are used in the research have unclear histological origin and probably do not represent HGSOC. Thus, new and reliable models of HGSOC are needed. Ascitic fluid from a patient with recurrent HGSOC was used to establish a stable cancer cell line. Cells were characterized by cytogenetic karyotyping and short tandem repeat (STR) profiling. New generation sequencing was applied to test for hot-spot mutations in 50 cancer-associated genes and fluorescence in situ hybridization (FISH) analysis was used to check for TP53 status. Cells were analyzed for expression of several marker genes/proteins by reverse-transcription polymerase chain reaction (RT-PCR), fluorescence-activated cell sorting (FACS), and immunocytochemistry (ICC). Functional tests were performed to compare OVPA8 cells with five commercially available and frequently used ovarian cancer cell lines: SKOV3, A2780, OVCAR3, ES2, and OAW42. Our newly-established OVPA8 cell line shows morphologic and genetic features consistent with HGSOC, such as epithelial morphology, multiple chromosomal aberrations, TP53 mutation, BRCA1 mutation, and loss of one copy of BRCA2. The OVPA8 line has a stable STR profile. Cells are positive for EpCAM, CK19, and CD44; they have relatively low plating efficiency/ability to form spheroids, a low migration rate, and intermediate invasiveness in matrigel, as compared to other ovarian cancer lines. OVPA8 is sensitive to paclitaxel and resistant to cisplatin. We also tested two FGFR inhibitors; OVPA8 cells were resistant to AZD4547 (AstraZeneca, London, UK), but sensitive to the new inhibitor CPL304-110-01 (Celon Pharma, Lomianki/Kielpin, Poland). We have established and characterized a novel cell line, OVPA8, which can be a valuable preclinical model for studies on high-grade serous ovarian cancer.
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Cistadenocarcinoma Seroso/patologia , Neoplasias Ovarianas/patologia , Proteína BRCA1 , Proteína BRCA2 , Linhagem Celular Tumoral , Aberrações Cromossômicas , Cistadenocarcinoma Seroso/genética , Cistadenocarcinoma Seroso/metabolismo , Feminino , Predisposição Genética para Doença/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Cariotipagem , Mutação , Gradação de Tumores , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Sequências de Repetição em Tandem/genética , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismoRESUMO
Ovarian cancer is the most frequent cause of deaths from among gynecologic malignancies. Due to its asymptomatic development the disease is frequently diagnosed at an advanced, incurable stages. Although ovarian cancers usually respond well to the first line chemotherapy based on platinum compounds and taxanes, majority of patients develop recurrence and chemo-resistance. Despite many years of studies there is still lack of reliable diagnostic markers as well as other diagnostic methods enabling early detection and suitable for screening. Thus, current studies are aimed on finding new biomarkers with diagnostic, prognostic and predictive potential as well as on the search for the new therapeutic targets. Interestingly, an understanding of ovarian cancer etiology has changed fundamentally within recent years. The classical theory, claiming that ovarian cancers originate from ovarian surface epithelial cells, was undermined. Currently, there is a lot of evidence that majority of serous ovarian cancers have its origin in malignant tubal epithelium, while endometrioid and clear cell ovarian cancers develop most likely from endometriosis. These new findings will have an impact on diagnostic approaches as well as on the prevention options for women with genetic predisposition to ovarian cancer. The new knowledge about an origin of different histological types of ovarian cancer may open new pathways in basic research and clinical studies. In this paper we report current knowledge about ovarian cancer risk factors, we also present the arguments for extraovarian origin of the majority of ovarian cancers and stress the mechanisms of action of new drugs for targeted therapies that show most promising results in the current clinical trials.
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Biomarcadores Tumorais , Neoplasias Ovarianas/etiologia , Feminino , Predisposição Genética para Doença , Humanos , Neoplasias Ovarianas/diagnóstico , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/terapiaRESUMO
BACKGROUND: The objectives of this study were to investigate the utility of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA)/endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) for the diagnosis of amyloidosis coupled with the feasibility of mass spectrometry (MS) for amyloid subtyping. METHODS: All patients who had amyloid diagnosed by EBUS-TBNA/EUS-FNA at two tertiary care centers from 2011 to 2020 were retrieved along with the MS subtype, clinical findings, and outcomes. RESULTS: Eight patients were included: seven underwent EBUS-TBNA of mediastinal lymph nodes, and one underwent EUS-FNA of a periportal lymph node. Ages ranged from 37 to 79 years (median, 69 years), with equal numbers of men and women. Presenting clinical history included one case each of follicular lymphoma, lymphoplasmacytic lymphoma, rheumatoid arthritis, possible sarcoid, cirrhosis, and chronic renal insufficiency, and one case each of suspected pulmonary and cardiac amyloidosis. All cases showed waxy, amorphous material on direct smears (n = 5) or ThinPrep slides (n = 3), which were confirmed as amyloid on Congo Red staining. Immunohistochemistry showed dominant lambda staining in two of three cases. MS was performed in all cases and identified five of the light-chain (AL) type, one of the heavy-chain/AL type, and two suggestive of AL amyloidosis. Bone marrow biopsy performed in seven patients demonstrated that three had monoclonal plasma cells and one had lymphoplasmacytic lymphoma. Two of four patients with systemic amyloidosis received chemotherapy and remained alive, whereas three with localized disease remained stable under observation. CONCLUSIONS: EBUS-TBNA/EUS-FNA is effective for amyloidosis diagnosis and provides adequate material for ancillary tests, including MS, which can identify the precursor amyloidogenic protein, leading to appropriate patient management.
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Amiloidose , Neoplasias Pulmonares , Linfoma , Masculino , Humanos , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Aspiração por Agulha Fina Guiada por Ultrassom Endoscópico/métodos , Centros de Atenção Terciária , Atenção Terciária à Saúde , Broncoscopia/métodos , Mediastino/diagnóstico por imagem , Mediastino/patologia , Neoplasias Pulmonares/patologia , Linfonodos/patologia , Amiloidose/diagnóstico , Amiloidose/etiologia , Amiloidose/patologia , Linfoma/patologia , Estadiamento de NeoplasiasRESUMO
Testing cognitive function in patients after severe brain damage is a major clinical challenge. In the absence of both verbal and motor communication, tests commonly used to assess cognitive function are completely or partially undoable for disorders of consciousness patients. The study involved 12 patients with varying degrees of impaired consciousness due to brain damage, with no verbal and motor communication. Memory was assessed in study participants using oculography. Memory tasks were presented in four categories. The total percentage of correctly completed tasks obtained across the group was 39.58%. The most difficult tasks included category C.4 with tasks involving working memory. Regardless of the subjects' level of consciousness, there was no statistically significant difference in the percentage of correct responses obtained in subgroups distinguished by CRS-R score. Eye tracking technology can be successfully used in the assessment of cognitive function, particularly when eye movements are the only channel of communication in individuals after brain damage. We suggest that the cognitive functions of people after brain damage should be further analyzed using eye tracking.
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BACKGROUND: The present investigation was designed to determine cognitive performance and quality of life (QoL) in a group of elite athletes who sustained spinal cord injury (SCI). METHODS: nine participants suffering a SCI participated in the study. Different cognitive functions were evaluated through the following tests: COWAT, Digit Span, Stroop color-word and QoL through the WHOQoL-BREF scale. RESULTS: Generally, participants positively assessed their overall quality of life and health status. Although the tests conducted indicate reduced cognitive function among the athletes, it did not affect the reduction in QoL. Single correlations between the results of cognitive tests and QoL could be treated as coincidental. CONCLUSIONS: Despite the observed decline in selected cognitive functions, the participants positively assessed their quality of life and physical health.Reduced cognitive functioning could be influenced by the impact of sleep-disordered breathing, pain, depressive disorders and medication. This indicates the need for an individualized approach to define the patient's deficits, needs and best care. Further studies with a larger group of participants are needed.
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Qualidade de Vida , Traumatismos da Medula Espinal , Atletas , Cognição , Humanos , Testes Neuropsicológicos , Qualidade de Vida/psicologia , Traumatismos da Medula Espinal/psicologiaRESUMO
Spinal cord injury (SCI) affects every aspect of human life: medical, psychological, social, material. People with SCI face a variety of secondary conditions (e.g., chronic pain, urinary tract infections, cognitive impairment) that place a significant emotional burden, resulting in an increased risk of depression and reduced quality of life. The purpose of this study was to better understand the coping strategies and to identify factors that promote or hinder the successful adjustment of elite athletes after SCI. Individual semi-structured interviews were conducted with eight top athletes after spinal cord injury. The interviews were recorded, transcribed, and then thematically analyzed using MAXQDA software. Thematic analysis identified the following categories: coping, athletic identity, and adjustment. The results of the study indicate that loss of functional ability does not cause loss of athlete identity. Elite athletes live a life consistent with this identity, attempting to maintain it despite the loss of physical fitness. Involvement in sports provides meaning and is a positive factor in the process of disability acceptance, which is essential in the process of adjustment to injury and also provides group belonging.
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Traumatismos da Medula Espinal , Esportes , Adaptação Psicológica , Atletas/psicologia , Humanos , Qualidade de Vida/psicologia , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/epidemiologiaRESUMO
The eyeballs are often the only way to communicate messages as a result of brain damage. However, it is not uncommon for them to become dysfunctional, thus requiring the introduction of appropriate therapy. The trajectory of eye movements (saccadic movements and gaze fixation) during observation of a static and dynamic point presented with an eye tracker was analyzed in the present study. Twelve patients with brain injury of different etiology, with different degrees of consciousness disorders and not communicating through verbal and motor skills, qualified for the study. All participants demonstrated greater eye movement activity when presented with a dynamic task in which they observed a moving point. The findings suggest that effective eye movement therapy must incorporate dynamic stimuli.
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Lesões Encefálicas , Movimentos Oculares , Encéfalo , Lesões Encefálicas/complicações , Estado de Consciência , Fixação Ocular , HumanosRESUMO
The quality of life of patients with severe brain damage is compromised by, e.g., impaired cognitive functions and ocular dysfunction. The paper contains research findings regarding participants of an oculomotor training course aimed at the therapy of visual-spatial functions. Five male patients with brain damage who did not communicate, verbally or motorically, participated in the study. Over a six-week period, the subjects solved tasks associated with recognising objects, size perception, colour perception, perception of object structures (letters), perception of object structures (objects), detecting differences between images and assembling image components into the complete image with the use of an eye tracker. The findings present evidence of oculomotor training effectiveness based on a longer duration of the work with the eye tracker and improved visual-spatial functions.
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Estado de Consciência , Qualidade de Vida , Percepção de Cores , Movimentos Oculares , Humanos , Masculino , Visão OcularRESUMO
BACKGROUND: Testing for tumor programmed death ligand-1 (PD-L1) expression was initially developed with histology specimens in non-small cell lung cancer (NSCLC). However, cytology specimens are widely used for primary diagnosis and biomarker studies in clinical practice. Limited clinical data exist on the predictiveness of cytology-derived PD-L1 scores for response to immune checkpoint inhibitor (ICI) therapy. METHODS: We reviewed all NSCLC specimens clinically tested at the University Health Network (UHN) for PD-L1 with 22C3pharmDx, from 01/2013 to 04/2021. Treatment outcomes in patients treated with single agent ICI therapy were reviewed and compared according to cytology- and histology-derived PD-L1 scores. RESULTS: We identified 494 and 1942 unique patients with cytology- and histology-derived tumor proportion scores, respectively, during the study period. Informative testing rates were 95 % vs 98 % for cytology and histology, respectively. Clinical data were available for 152 patients treated with single agent ICI: 61 cytology and 91 histology. Overall response rates (ORR) were similar for cytology and histology (36 % vs 34 %; p = 0.23), as well as median progression free survival (PFS) (4.9 vs 4.2 months; p = 0.99) and overall survival (23.4 vs 19.7 months; p = 0.99). The results remained similar even after adjusting for PD-L1 expression levels and line of ICI treatment (PFS HR 1.15; 95 %CI 0.78-1.70; p = 0.47). CONCLUSIONS: Treatment outcomes to single agent ICI based on cytology-derived PD-L1 scores were comparable to histology controls. Our results support PD-L1 biomarker testing on both cytology and histology specimens.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Antígeno B7-H1/metabolismo , Biomarcadores Tumorais/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Humanos , Neoplasias Pulmonares/patologiaRESUMO
This paper describes the synthesis and antiproliferative activity of conjugates of genistein (1) and unsaturated pyranosides. Constructs linking genistein with a sugar moiety through an alkyl chain were obtained in a two-step synthesis: in a first step genistein was converted into an intermediate bearing an ω-hydroxyalkyl substituent, containing two, three or five carbon atoms, at position 7, while the second step involved Ferrier glycosylation reaction, employing glycals. Antiproliferative activity of several genistein derivatives was tested in cancer cell lines in vitro. The most potent derivative, Ram-3 inhibited the cell cycle, interacted with mitotic spindles and caused apoptotic cell death. Neither genistein nor the sugar alone were able to influence the mitotic spindle organization. Our results indicate, that conjugation of genistein with certain sugars may render the interaction of derivatives with new molecular targets.
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Proliferação de Células/efeitos dos fármacos , Genisteína/farmacologia , Mitose/efeitos dos fármacos , Linhagem Celular Tumoral , Eletroforese em Gel de Ágar , Genisteína/síntese química , Humanos , Marcação In Situ das Extremidades Cortadas , Espectroscopia de Ressonância Magnética , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
This study aimed to describe the percentage of tasks involving language functions that were completed by patients diagnosed with disorders of consciousness, as observed during neurorehabilitation conducted for different periods of time using an alternative communication tool. The project involved six participants, who were observed for 1 month, 6 months, and 1 year. The patients were asked to solve tasks involving language functions with the use of an eye-controlled device. The language functions were evaluated on the basis of the average number of tasks performed by the patients, which was 70.45% for the whole subject group. It is not entirely clear what determined the changes in language functions during the research. It is crucial that patients performed the presented tasks even though their state of consciousness, as confirmed through medical documentation (unresponsive wakefulness syndrome), did not suggest the possibility of establishing any contact with them.
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BACKGROUND: A spinal cord injury (SCI) is a traumatic event that affects every aspect of life: physical, mental, economic, and social. The main aim of this study was to investigate self-efficacy, quality of life, and their correlations among outstanding athletes who have suffered spinal cord injuries, and to determine whether these individuals have specific psychological characteristics that contribute to a better quality of life. METHODS: The study involved nine athletes with at least national-level achievements in sports prior to an SCI. Participation in the study consisted of an interview via an online communicator, followed by an online questionnaire consisting of a personal questionnaire and two scales: The World Health Organization Quality of Life Scale (WHOQoL-BREF), and the General Self-Efficacy Scale (GSES). RESULTS: Spearman's correlation showed a correlation between general self-efficacy, perception of quality of life, and satisfaction with own physical health, as well as psychological resources and environmental support. CONCLUSIONS: Involvement in an environment that was important to the injured person before the accident, in either a passive (in the absence of functional capacity) or active form, promotes a greater sense of self-efficacy and good QoL, regardless of the time that has passed since the accident, and despite high levels of pain or secondary health issues. To fill the gap in professional long-term healthcare services for athletes after SCIs, intervention programs should be considered that support self-efficacy, which is an important factor that can be subject to improvement.
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Traumatismos da Medula Espinal , Esportes , Atletas , Humanos , Qualidade de Vida , Autoeficácia , Inquéritos e QuestionáriosRESUMO
FGFR signalling is one of the most prominent pathways involved in cell growth and development as well as cancer progression. FGFR1 amplification occurs in approximately 20% of all squamous cell lung carcinomas (SCC), a predominant subtype of non-small cell lung carcinoma (NSCLC), indicating FGFR as a potential target for the new anti-cancer treatment. However, acquired resistance to this type of therapies remains a serious clinical challenge. Here, we investigated the NSCLC cell lines response and potential mechanism of acquired resistance to novel selective FGFR inhibitor CPL304110. We found that despite significant genomic differences between CPL304110-sensitive cell lines, their resistant variants were characterised by upregulated p38 expression/phosphorylation, as well as enhanced expression of genes involved in MAPK signalling. We revealed that p38 inhibition restored sensitivity to CPL304110 in these cells. Moreover, the overexpression of this kinase in parental cells led to impaired response to FGFR inhibition, thus confirming that p38 MAPK is a driver of resistance to a novel FGFR inhibitor. Taken together, our results provide an insight into the potential direction for NSCLC targeted therapy.
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Carcinoma Pulmonar de Células não Pequenas/enzimologia , Carcinoma Pulmonar de Células não Pequenas/patologia , Resistencia a Medicamentos Antineoplásicos , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/patologia , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Biomarcadores Tumorais/metabolismo , Pontos de Checagem do Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , Humanos , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismoRESUMO
A three-times World Champion in BMX (an acronym for Bicycle Motocross) dirt jumps, a Junior World Champion in ski jumping, and a European karate Champion sustained spinal cord injuries at the cervical and thoracic level. Such a severe trauma is tantamount to the end of a professional sporting career. In such a situation, the athlete's life significantly changes in every aspect of it: health, professional, and social. The greatest sports champions have not yet been portrayed in the context of a strategy they used to deal with an abrupt end of a professional career due to severe injury. A semi-structured interview was conducted with study participants who additionally filled out the WHO Quality of Life Scale. This multiple case series presents the quality of life in elite athletes as well as the social activities they have undertaken regardless of the tragic accident. The results of the research indicate that these people are characterized rather by a positive sense of quality in life, and the way they function in a difficult situation is an inspiration to others.
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Atletas/psicologia , Traumatismos em Atletas/etiologia , Vértebras Cervicais/lesões , Qualidade de Vida/psicologia , Traumatismos da Medula Espinal/psicologia , Esportes , Vértebras Torácicas/lesões , Adulto , Traumatismos em Atletas/diagnóstico , Humanos , Fatores de RiscoRESUMO
Previously, based on a DNA microarray experiment, we identified a 96-gene prognostic signature associated with the shorter survival of ovarian cancer patients. We hypothesized that some differentially expressed protein-coding genes from this signature could potentially serve as prognostic markers. The present study was aimed to validate two proteins, namely fibronectin (FN1) and periostin (POSTN), in the independent set of ovarian cancer samples. Both proteins are mainly known as extracellular matrix proteins with many important functions in physiology. However, there are also indications that they are implicated in cancer, including ovarian cancer. The expression of these proteins was immunohistochemically analyzed in 108 surgical samples of advanced ovarian cancer (majority: high-grade serous) and additionally on tissue arrays representing different stages of the progression of ovarian and fallopian tube epithelial tumors, from normal epithelia, through benign tumors, to adenocarcinomas of different stages. The correlation with clinical, pathological, and molecular features was evaluated. Kaplan-Meier survival analysis and Cox-proportional hazards models were used to estimate the correlation of the expression levels these proteins with survival. We observed that the higher expression of fibronectin in the tumor stroma was highly associated with shorter overall survival (OS) (Kaplan-Meier analysis, log-rank test p = 0.003). Periostin was also associated with shorter OS (p = 0.04). When we analyzed the combined score, calculated by adding together individual scores for stromal fibronectin and periostin expression, Cox regression demonstrated that this joint FN1&POSTN score was an independent prognostic factor for OS (HR = 2.16; 95% CI: 1.02-4.60; p = 0.044). The expression of fibronectin and periostin was also associated with the source of ovarian tumor sample: metastases showed higher expression of these proteins than primary tumor samples (χ2 test, p = 0.024 and p = 0.032). Elevated expression of fibronectin and periostin was also more common in fallopian cancers than in ovarian cancers. Our results support some previous observations that fibronectin and periostin have a prognostic significance in ovarian cancer. In addition, we propose the joint FN1&POSTN score as an independent prognostic factor for OS. Based on our results, it may also be speculated that these proteins are related to tumor progression and/or may indicate fallopian-epithelial origin of the tumor.
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Adenocarcinoma Mucinoso/secundário , Biomarcadores Tumorais/metabolismo , Moléculas de Adesão Celular/metabolismo , Cistadenocarcinoma Seroso/secundário , Neoplasias do Endométrio/secundário , Fibronectinas/metabolismo , Neoplasias Ovarianas/patologia , Adenocarcinoma Mucinoso/tratamento farmacológico , Adenocarcinoma Mucinoso/metabolismo , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Cistadenocarcinoma Seroso/tratamento farmacológico , Cistadenocarcinoma Seroso/metabolismo , Neoplasias do Endométrio/tratamento farmacológico , Neoplasias do Endométrio/metabolismo , Feminino , Seguimentos , Humanos , Pessoa de Meia-Idade , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/metabolismo , Prognóstico , Taxa de SobrevidaRESUMO
In our previous microarray study we identified two subgroups of high-grade serous ovarian cancers with distinct gene expression and survival. Among differentially expressed genes was an Integrin beta-like 1 (ITGBL1), coding for a poorly characterized protein comprised of ten EGF-like repeats. Here, we have analyzed the influence of ITGBL1 on the phenotype of ovarian cancer (OC) cells. We analyzed expression of four putative ITGBL1 mRNA isoforms in five OC cell lines. OAW42 and SKOV3, having the lowest level of any ITGBL1 mRNA, were chosen to produce ITGBL1-overexpressing variants. In these cells, abundant ITGBL1 mRNA expression could be detected by RT-PCR. Immunodetection was successful only in the culture media, suggesting that ITGBL1 is efficiently secreted. We found that ITGBL1 overexpression affected cellular adhesion, migration and invasiveness, while it had no effect on proliferation rate and the cell cycle. ITGBL1-overexpressing cells were significantly more resistant to cisplatin and paclitaxel, major drugs used in OC treatment. Global gene expression analysis revealed that signaling pathways affected by ITGBL1 overexpression were mostly those related to extracellular matrix organization and function, integrin signaling, focal adhesion, cellular communication and motility; these results were consistent with the findings of our functional studies. Overall, our results indicate that higher expression of ITGBL1 in OC is associated with features that may worsen clinical course of the disease.
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BACKGROUND: Programmed death ligand-1 (PD-L1) assessed by immunohistochemistry (IHC) is used as biomarker for pembrolizumab therapy in advanced stage lung cancer patients. However, data permitting direct performance comparison between cytology and surgical specimen types are limited since both specimens from a single tumor site are infrequently available. In addition, alcohol fixation used with cytology specimens requires technical validation of the PD-L1 IHC assay before clinical use. We here report our experience with implementation of the PD-L1 22C3 IHC pharmDxTM assay for cytologic samples at a large tertiary cancer center. STUDY DESIGN: Archival formalin-fixed (FF), paraffin-embedded cell blocks (CBs) and subsequent lung tumor resections (LTRs) from the same anatomical site were used for a direct comparison of PD-L1 tumor proportion scores (TPSs). TPS values were independently determined by one surgical lung pathologist and two cytopathologists blinded to the specimen pairs. An interim analysis was performed to facilitate the pooling of expertise among observers. After PD-L1 22C3 IHC pharmDxTM implementation for FF cytology specimens, dual-processed samples were used for a prospective technical validation of CytoLyt® prefixation (CF). Digital image analysis was performed for a subset of dual-processed specimens. RESULTS: Eighty-one CBs and LTRs were included for comparison of the specimen types. PD-L1 assessment in CBs had an accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of 88.9/72.8, 66.7/73.5, 95.2/72.3, 80.0/65.8, and 90.9/79.1% for the ≥50/≥1% cutoff, respectively. The intraclass correlation coefficient was 0.84 (95% confidence interval [CI]: 0.76, 0.90), and it improved after interim analysis (before: 0.79 and after: 0.92). The overall concordance between CF and FF for the categories defined by the ≥50/≥1% cutoff values was 90.4% (95% CI: 79.0, 96.8). Similar assay performance was confirmed by digital analysis. CONCLUSIONS: PD-L1 22C3 IHC pharmDxTM shows good reliability if used with CB preparations. CF does not impact assay results significantly. Clinical validation with outcome data is needed, and digital methods of assessment should be further investigated.
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Antígeno B7-H1/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Imuno-Histoquímica , Adulto , Anticorpos Monoclonais Humanizados/farmacologia , Antígeno B7-H1/análise , Biomarcadores Tumorais/análise , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Feminino , Humanos , Imuno-Histoquímica/métodos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos TestesRESUMO
An understanding of the molecular pathogenesis and heterogeneity of ovarian cancer holds promise for the development of early detection strategies and novel, efficient therapies. In this review, we discuss the advantages and limitations of animal models available for basic and preclinical studies. The fruit fly model is suitable mainly for basic research on cellular migration, invasiveness, adhesion, and the epithelial-to-mesenchymal transition. Higher-animal models allow to recapitulate the architecture and microenvironment of the tumor. We discuss a syngeneic mice model and the patient derived xenograft model (PDX), both useful for preclinical studies. Conditional knock-in and knock-out methodology allows to manipulate selected genes at a given time and in a certain tissue. Such models have built our knowledge about tumor-initiating genetic events and cell-of-origin of ovarian cancers; it has been shown that high-grade serous ovarian cancer may be initiated in both the ovarian surface and tubal epithelium. It is postulated that clawed frog models could be developed, enabling studies on tumor immunity and anticancer immune response. In laying hen, ovarian cancer develops spontaneously, which provides the opportunity to study the genetic, biochemical, and environmental risk factors, as well as tumor initiation, progression, and histological origin; this model can also be used for drug testing. The chick embryo chorioallantoic membrane is another attractive model and allows the study of drug response.
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Based on results of pilot scale research with source-separated black water (BW) and grey water (GW), a new sanitation concept is proposed. BW and GW are both treated in a UASB (-septic tank) for recovery of CH4 gas. Kitchen waste is added to the anaerobic BW treatment for doubling the biogas production. Post-treatment of the effluent is providing recovery of phosphorus and removal of remaining COD and nitrogen. The total energy saving of the new sanitation concept amounts to 200 MJ/year in comparison with conventional sanitation, moreover 0.14 kg P/p/year and 90 litres of potential reusable water are produced.