RESUMEN
Oral cancer is a leading cause of cancerrelated death worldwide. Current treatment for oral cancer includes surgery, radiotherapy, and chemotherapy; however, their effectiveness is still limited. To identify a new prognostic biomarker and therapeutic target for oral cancer, the Opa interacting protein 5 (OIP5), which plays an essential role in the proper segregation of chromosomes, was examined. Immunohistochemical staining using tissue microarrays indicated that OIP5 was expressed in 120 of 164 (73.2%) oral cancers but was minimally expressed in normal oral tissues. OIP5 expression was significantly associated with poor prognosis in patients with oral cancer. Overexpression of OIP5 enhanced the growth of oral cancer cells, whereas OIP5 knockdown using small interfering RNAs (siRNAs) significantly inhibited cell growth through cell cycle arrest at the G2/M phase. Suppression of OIP5 expression also induced senescence of oral cancer cells. Overall, the findings of the present study suggest that OIP5 may be a candidate prognostic biomarker and therapeutic target in oral cancer.
Asunto(s)
Proteínas de Ciclo Celular/análisis , Proteínas Cromosómicas no Histona/análisis , Neoplasias de la Boca/tratamiento farmacológico , Análisis de Varianza , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/sangre , Proteínas de Ciclo Celular/sangre , Proteínas de Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral/efectos de los fármacos , Proliferación Celular/genética , Proteínas Cromosómicas no Histona/sangre , Proteínas Cromosómicas no Histona/efectos de los fármacos , Humanos , Neoplasias de la Boca/fisiopatologíaRESUMEN
An initiating DNA double strand break (DSB) event precedes the formation of cancer-driven chromosomal abnormalities, such as gene rearrangements. Therefore, measuring DNA breaks at rearrangement-participating regions can provide a unique tool to identify and characterize susceptible individuals. Here, we developed a highly sensitive and low-input DNA break mapping method, the first of its kind for patient samples. We then measured genome-wide DNA breakage in normal cells of acute myeloid leukemia (AML) patients with KMT2A (previously MLL) rearrangements, compared to that of nonfusion AML individuals, as a means to evaluate individual susceptibility to gene rearrangements. DNA breakage at the KMT2A gene region was significantly greater in fusion-driven remission individuals, as compared to nonfusion individuals. Moreover, we identified select topoisomerase II (TOP2)-sensitive and CCCTC-binding factor (CTCF)/cohesin-binding sites with preferential DNA breakage in fusion-driven patients. Importantly, measuring DSBs at these sites, in addition to the KMT2A gene region, provided greater predictive power when assessing individual break susceptibility. We also demonstrated that low-dose etoposide exposure further elevated DNA breakage at these regions in fusion-driven AML patients, but not in nonfusion patients, indicating that these sites are preferentially sensitive to TOP2 activity in fusion-driven AML patients. These results support that mapping of DSBs in patients enables discovery of novel break-prone regions and monitoring of individuals susceptible to chromosomal abnormalities, and thus cancer. This will build the foundation for early detection of cancer-susceptible individuals, as well as those preferentially susceptible to therapy-related malignancies caused by treatment with TOP2 poisons.
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Factor de Unión a CCCTC/genética , ADN-Topoisomerasas de Tipo II/genética , N-Metiltransferasa de Histona-Lisina/genética , Leucemia Mieloide Aguda/genética , Proteína de la Leucemia Mieloide-Linfoide/genética , Proteínas de Unión a Poli-ADP-Ribosa/genética , Sitios de Unión/genética , Factor de Unión a CCCTC/sangre , Proteínas de Ciclo Celular/sangre , Proteínas de Ciclo Celular/genética , Proteoglicanos Tipo Condroitín Sulfato/sangre , Proteoglicanos Tipo Condroitín Sulfato/genética , Proteínas Cromosómicas no Histona/sangre , Proteínas Cromosómicas no Histona/genética , Aberraciones Cromosómicas , Roturas del ADN de Doble Cadena/efectos de los fármacos , Reparación del ADN/genética , ADN-Topoisomerasas de Tipo II/sangre , Proteínas de Unión al ADN/sangre , Proteínas de Unión al ADN/genética , Etopósido/farmacología , Femenino , Reordenamiento Génico/genética , Genoma Humano/genética , Células HeLa , N-Metiltransferasa de Histona-Lisina/sangre , Humanos , Leucemia Mieloide Aguda/sangre , Leucemia Mieloide Aguda/patología , Masculino , Proteína de la Leucemia Mieloide-Linfoide/sangre , Proteínas de Fusión Oncogénica/genética , Proteínas de Unión a Poli-ADP-Ribosa/sangre , CohesinasRESUMEN
The nuclear envelope component proline-rich protein 14 (PRR14) is involved in the nuclear morphological alteration and activation of the mTOR (mammalian target of rapamycin) signaling pathway, and has been repeatedly shown to be upregulated in patients with Parkinson's disease (PD). The aim of this study was to explore whether PRR14 can be used as a potential biomarker for the diagnosis of PD. We compared PRR14 expression in PD patients and normal controls in gene expression omnibus (GEO) data. Quantitative enzyme-linked immunosorbent assay (ELISA) was used to detect PRR14 expression in PD patients and age- and sex-matched controls. The relationship between serum PRR14 and clinical phenotype was evaluated using correlation analysis and logistic regression. The expression of PRR14 in whole blood, substantia nigra, and medial substantia nigra was significantly higher in PD patients than in the healthy control group. Compared to plasma, serum was more suitable for the detection of PRR14. Furthermore, serum PRR14 level in PD patients was significantly higher than that in age- and sex-matched controls. The area under the curve for serum PRR14 level in the ability to identify PD versus age- and sex-matched controls was 0.786. In addition, serum PRR14 level was found to correlate with constipation in PD patients. Our findings demonstrate for the first time that serum PRR14 is a potential biomarker for PD.
Asunto(s)
Proteínas Cromosómicas no Histona/sangre , Proteínas del Tejido Nervioso/sangre , Enfermedad de Parkinson/diagnóstico , Anciano , Biomarcadores , Estudios de Casos y Controles , Proteínas Cromosómicas no Histona/biosíntesis , Proteínas Cromosómicas no Histona/genética , Estreñimiento/sangre , Estreñimiento/etiología , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proteínas del Tejido Nervioso/biosíntesis , Proteínas del Tejido Nervioso/genética , Enfermedad de Parkinson/complicaciones , Enfermedad de Parkinson/metabolismo , Fenotipo , Plasma , Curva ROC , Sensibilidad y Especificidad , Suero , Transducción de Señal , Sustancia Negra/metabolismo , Evaluación de Síntomas , Serina-Treonina Quinasas TOR/fisiología , Regulación hacia ArribaRESUMEN
BACKGROUND: We hypothesized that the SKA2 gene can convert hemoglobin F to A leading to the maturity of the hematopoietic system by glucocorticoid hormone; so, the present study aimed to investigate the health outcome of newborns by using the effect of SKA2 gene on hematopoietic maturation. METHODS: At first, 142 samples were divided into term and preterm. After sampling from the umbilical cord blood, the expression of SKA2 genes and HbA and F were evaluated by quantitative RT-PCR. The blood gases were measured by Campact 3 device. Finally, the cortisol level was measured by ELISA method and HbA and F levels were investigated by capillary electrophoresis. RESULTS: The blood gases and Apgar scores were more favorable in term newborns (P <0.001). Levels of protein/expression of HbF in newborns with Apgar score greater than 7 was lower than that of the newborns with Apgar score below 7 (P <0.001). Cortisol and HbA levels were considerably higher in term newborns compared to the preterm ones (P <0.001). In the preterm and term groups, SKA2 gene expression had a positive and significant relationship with cortisol and HbA levels as well as a negative relationship with the HbF level. In the preterm group, a positive and significant relationship was observed between the expression of SKA2 and HbF genes. CONCLUSION: The results revealed that the SKA2 gene affected hematopoietic maturation in preterm and term newborns and the health outcome of newborns improved by increasing HbA level.
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Proteínas Cromosómicas no Histona/sangre , Hemoglobina Fetal/metabolismo , Hematopoyesis , Células Madre Hematopoyéticas/metabolismo , Hemoglobina A/metabolismo , Hidrocortisona/sangre , Recien Nacido Prematuro , Proteínas Cromosómicas no Histona/genética , Sangre Fetal/metabolismo , Hemoglobina Fetal/genética , Edad Gestacional , Hemoglobina A/genética , Humanos , Recién Nacido , Nacimiento a TérminoRESUMEN
Cornelia de Lange syndrome Spectrum (CdLSp) is characterized by intellectual disability, facial dysmorphisms, and growth impairment. Although eating difficulties are a well-known feature of the disease, there is no data regarding the nutritional deficiencies of these patients. The food intake was tracked using a dietary transcription provided by the family/caregivers, biochemical nutritional parameters were measured with laboratory tests and through an accurate clinical evaluation of the incidence of qualitative and quantitative imbalances in a cohort of 73 patients with CdLSp ware determined. Of these 73, 62 (85%) subjects provided a complete and detailed dietary transcription. In the studied population, a quantitative caloric imbalance in 47/62 (76%) subjects was observed. The caloric intake was low in 27/62 (43%) subjects whereas excessive in 20/62 (33%). Only 15/62 (24%) had an optimum caloric intake. Regarding micronutrients, a calcium intake deficiency in 32% of the patients (20/62) was observed. Blood tests revealed a low iron level in 22/73 (30%) of the patients and 25(OH)D deficiency in 49/73 (67%). Serum hypocalcemia was not evidenced. Qualitative and quantitative imbalances resulted in more frequent than expected in CdLSp patients. A qualitative imbalance was more prevalent in younger patients while in older patients prevailed mainly a quantitative disproportion. We found no statistically meaningful correlation between dietary imbalances, genetic, or clinical parameters. Our findings highlight the need for further studies to evaluate the basal metabolic rate of CdLSp patients and find a correlation with their growth impairment.
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Síndrome de Cornelia de Lange/genética , Ingestión de Alimentos/genética , Discapacidad Intelectual/genética , Desnutrición/genética , Adolescente , Proteínas de Ciclo Celular/sangre , Niño , Preescolar , Proteínas Cromosómicas no Histona/sangre , Estudios de Cohortes , Síndrome de Cornelia de Lange/sangre , Síndrome de Cornelia de Lange/metabolismo , Síndrome de Cornelia de Lange/patología , Femenino , Humanos , Discapacidad Intelectual/sangre , Discapacidad Intelectual/metabolismo , Discapacidad Intelectual/patología , Hierro/sangre , Italia , Masculino , Desnutrición/sangre , Desnutrición/metabolismo , Desnutrición/patología , FenotipoRESUMEN
Gastric cancer (GC) is the second most widespread cause of cancer-related mortality worldwide. The discovery of novel biomarkers of oncoproteins can facilitate the development of therapeutic strategies for GC treatment. In this study, we identified novel biomarkers by integrating isobaric tags for relative and absolute quantitation (iTRAQ), a human plasma proteome database, and public Oncomine datasets to search for aberrantly expressed oncogene-associated proteins in GC tissues and plasma. One of the most significantly upregulated biomarkers, DEK, was selected and its expression validated. Our immunohistochemistry (IHC) (n = 92) and quantitative real-time polymerase chain reaction (qRT-PCR) (n = 72) analyses disclosed a marked increase in DEK expression in tumor tissue, compared with paired nontumor mucosa. Importantly, significantly higher preoperative plasma DEK levels were detected in GC patients than in healthy controls via enzyme-linked immunosorbent assay (ELISA). In clinicopathological analysis, higher expression of DEK in both tissue and plasma was significantly associated with advanced stage and poorer survival outcomes of GC patients. Data from receiver operating characteristic (ROC) curve analysis disclosed a better diagnostic accuracy of plasma DEK than carcinoembryonic antigen (CEA), carbohydrate antigen 19.9 (CA 19.9), and C-reactive protein (CRP), highlighting its potential as an effective plasma biomarker for GC. Plasma DEK is also more sensitive in tumor detection than the other three biomarkers. Knockdown of DEK resulted in inhibition of GC cell migration via a mechanism involving modulation of matrix metalloproteinase MMP-2/MMP-9 level and vice versa. Our results collectively support plasma DEK as a useful biomarker for making diagnosis and prognosis of GC patients.
Asunto(s)
Proteínas Cromosómicas no Histona/análisis , Proteínas Oncogénicas/análisis , Proteínas de Unión a Poli-ADP-Ribosa/análisis , Neoplasias Gástricas/patología , Anciano , Biomarcadores de Tumor/análisis , Línea Celular Tumoral , Movimiento Celular , Proteínas Cromosómicas no Histona/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Metástasis de la Neoplasia/diagnóstico , Metástasis de la Neoplasia/patología , Proteínas Oncogénicas/sangre , Proteínas de Unión a Poli-ADP-Ribosa/sangre , Pronóstico , Neoplasias Gástricas/sangre , Neoplasias Gástricas/diagnóstico , Análisis de SupervivenciaRESUMEN
OBJECTIVE: Circular RNAs (circRNAs) have emerged as crucial regulators for tumor progression. However, the effects of circRNAs are not entirely clear in cervical cancer (CC). The objective of this study was to investigate the function and regulation mechanism of circular RNA SMARCA5 (circSMARCA5, also named hsa_circ_0001445) in cervical cancer. PATIENTS AND METHODS: circSMARCA5 and miR-620 expressions were analyzed by qRT-PCR assay. The cell proliferation, cell cycle, migration, invasion abilities were measured by CCK-8 and colony formation, Flow cytometry, Wound-healing and transwell assays. In addition, the interaction between circSMARCA5 and miR-620 was accessed by luciferase reporter assay. RESULTS: The results indicated that circSMARCA5 was downregulated in CC; overexpression of circSMARCA5 inhibited CC cell proliferation, migration and invasion, and induced cell cycle arrest. In addition, we found that circSMARCA5 could bind to miR-620 and significantly downregulated its expression. Furthermore, the results revealed that circSMARCA5 suppressed proliferation and invasion by miR-620 in CC. CONCLUSIONS: We suggested that circSMARCA5/miR-620 regulatory axis is involved in the development of CC and may serve as a potential therapy target.
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Adenosina Trifosfatasas/fisiología , Proteínas Cromosómicas no Histona/fisiología , MicroARNs/fisiología , ARN Circular , Neoplasias del Cuello Uterino/patología , Adenosina Trifosfatasas/sangre , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Proteínas Cromosómicas no Histona/sangre , Femenino , Humanos , Invasividad NeoplásicaRESUMEN
OBJECTIVE: To investigate the dynamic variation in H3K4me3 and HP1 with employment length in nickel smelting workers. METHODS: Blood samples were collected from 140 nickel smelting workers and 140 age-matched office workers to test for H3K4me3, and HP1 levels. RESULTS: H3K4me3 was statistically significantly different (p < 0.05) between the two groups and positively correlated with employment length (rs = 0.267). HP1 was not correlated with employment length (p = 0.066) but was significantly different between the two groups. CONCLUSIONS: Chronic exposure to nickel can induce oxidative damage, and increase H3K4me3 expression and inhibit HP1 expression.
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Metalurgia , Níquel/farmacología , Adulto , Estudios de Casos y Controles , Homólogo de la Proteína Chromobox 5 , Proteínas Cromosómicas no Histona/sangre , Histonas/sangre , Humanos , Níquel/orina , Exposición Profesional/efectos adversos , Exposición Profesional/análisis , Estrés Oxidativo/efectos de los fármacos , Factores de TiempoRESUMEN
Patients with systemic sclerosis (SSc) who express autoantibodies to centromeric proteins (CENPs) are at risk of developing pulmonary vascular disease and pulmonary arterial hypertension without fibrosis. Currently no biomarkers are available to predict these complications. We previously characterized the fine specificity of anti-CENP-A antibodies in SSc by screening a phage display library (expressing random 12-mer peptides), and identified phage clones whose peptides were differentially recognized by patients' autoantibodies. Here, we examined if subgroups of SSc patients with different anti-CENP-A antibody subspecificities also differ clinically, and if serum reactivity to phage-displayed peptides can predict pulmonary vascular disease.Clinical data and serum samples were collected from 84 anti-CENP-A-positive SSc patients. Indirect ELISAs were used to test serum reactivity. Pulmonary vascular disease was defined as high systolic pulmonary arterial pressure (sPAP) and low diffusing lung capacity for carbon monoxide (DLCO; percent of predicted values).Sera were screened for reactivity to peptides expressed by phage clones pc4.2 and pc14.1, confirming our earlier observation of differential specificities. Linear regression showed that the levels of antibodies specific for the 2 phage clones were associated with clinical features of pulmonary vascular disease, but in opposite ways: anti-pc4.2 antibodies were positively associated with sPAP and inversely associated with DLCO, whereas anti-pc14.1 antibodies were inversely associated with sPAP and positively associated with DLCO. Anti-pc4.2 and anti-pc14.1 antibody levels predicted sPAP independently of DLCO. These associations were confirmed by logistic regression using antibodies as predictors and dichotomized sPAP (cutoff, 45âmmâHg) as outcome. The ratio of the 2 antibody levels was a useful marker in predicting high sPAP.This study demonstrates that some SSc clinical features associate with subspecificities of anti-CENP-A antibodies. Moreover, it shows that a simple, inexpensive phage-based assay can predict which SSc patients have high sPAP and low DLCO, hence who are at greater risk of developing pulmonary arterial hypertension. The ability to identify these at-risk patients can contribute to clinical efficiency and effectiveness. Further research into the peptides expressed by the phage clones may reveal the molecular mechanisms that put some anti-CENP-A-positive patients at greater risk than others for pulmonary vascular disease.
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Autoanticuerpos/inmunología , Autoantígenos/inmunología , Autoinmunidad , Proteínas Cromosómicas no Histona/inmunología , Hipertensión Pulmonar/etiología , Esclerodermia Sistémica/complicaciones , Autoanticuerpos/sangre , Autoantígenos/sangre , Biomarcadores/sangre , Proteína A Centromérica , Proteínas Cromosómicas no Histona/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Estudios de Seguimiento , Humanos , Hipertensión Pulmonar/diagnóstico , Hipertensión Pulmonar/inmunología , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Esclerodermia Sistémica/sangre , Esclerodermia Sistémica/inmunologíaRESUMEN
The autoantibody profiles in New Zealand systemic sclerosis patients have not previously been reported. The aim of this study was to evaluate the autoantibody profiles of patients in the Waikato Hospital Systemic Sclerosis Clinic cohort. The EUROLINE (IgG) Systemic Sclerosis panel test kit (which tests for Scl-70, CENP-A, CENP-B, RP11, RP155, Fib, NOR90, Th/To, PM100, PM75, Ku, PDGFR and Ro-52) was selected for the purpose of this study. All patients attending the Waikato Hospital Systemic Sclerosis clinic were invited to participate. These patients were categorised by systemic sclerosis subtypes [1]. Results were compared with previously published data, including the EUSTAR database. Sixty patients (56 female) were recruited, with a median age of 61 years (range 29-81 years). Forty-one had limited cutaneous systemic sclerosis (lcSSc). Of these lcSSc patients, 31 (75.6%) were positive for CENP-A and CENP-B (anti-centromere) antibodies, 12 (29.3%) for Ro-52 antibodies, 5 (12.2%) for RP11 and RP155, 4 (9.8%) for Scl-70 and 1 (2.4%) each for anti-Fib and Th/To antibodies. Fifteen patients had diffuse cutaneous systemic sclerosis (dcSSc), of which 7 patients (47.6%) were positive for RP11 and RP155, 4 (26.7%) for Scl-70. Three dcSSc patients did not have either of these two major antibodies, but of these 15 dcSSc patients, 4 patients (26.7%) were positive also for Ro-52, 2 (13.3%) for anti-Ku, and 1 (6.7%) each for anti-Fib and NOR90. Four patients had overlap syndrome (OLS), 1 had CENP-A and CENP-B antibodies, 1 had Ro-52 autoantibodies 1 had anti-Ku antibodies. Three patients had no autoantibodies. This is the first study to look at the autoantibody profile of SSc patients in New Zealand. A higher prevalence of antibodies against centromere and RNA polymerase III was demonstrated in our group compared with the EUSTAR database suggesting that antibody prevalence may vary geographically.
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Anticuerpos Antinucleares/sangre , Autoanticuerpos/sangre , Autoantígenos/sangre , Proteínas Cromosómicas no Histona/sangre , Esclerodermia Sistémica/clasificación , Esclerodermia Sistémica/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Proteína A Centromérica , Estudios de Cohortes , Estudios Transversales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Nueva Zelanda , ARN Polimerasa III/sangreRESUMEN
Blood biomarkers may provide a scientifically useful and clinically usable peripheral signal in psychiatry, as they have been doing for other fields of medicine. Jumping to premature conclusions, negative or positive, can create confusion in this field. Reproducibility is a hallmark of good science. We discuss some recent examples from this dynamic field, and show some new data in support of previously published biomarkers for suicidality (SAT1, MARCKS and SKA2). Methodological clarity and rigor in terms of biomarker discovery, validation and testing is needed. We propose a set of principles for what constitutes a good biomarker, similar in spirit to the Koch postulates used at the birth of the field of infectious diseases.
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Biomarcadores/sangre , Trastorno Bipolar/sangre , Acetiltransferasas/sangre , Trastorno Bipolar/diagnóstico , Proteínas Cromosómicas no Histona/sangre , Femenino , Humanos , Péptidos y Proteínas de Señalización Intracelular/sangre , Masculino , Proteínas de la Membrana/sangre , Persona de Mediana Edad , Sustrato de la Proteína Quinasa C Rico en Alanina Miristoilada , SuicidioRESUMEN
INTRODUCTION: The systemic sclerosis-associated autoantibodies include anti-centromere, anti-topoisomerase I (anti-topo I), anti-RNA polymerase III, anti-fibrillarin, anti-Th/To, and anti-PDGFR. A specific serological profile is connected with clinical manifestations and prognosis in systemic sclerosis (SSc). OBJECTIVES: The aim of the study was to assess the serological profile in limited cutaneous and diffuse cutaneous SSc (lcSSc and dcSSc). PATIENTS AND METHODS: 87 (68 female and 19 male) consecutive SSc patients treated between 2006 and 2011 were assessed. Patients fulfilled the American College of Rheumatology classification criteria of SSc: 35 - dcSSc and 52 - lcSSc. The following marker antibodies were determined: anti-topo I, anti-centromere A and B (CENP A, CENP B), anti-RNA polymerase III (RP11, RP 155), anti-fibrillarin (U3RNP), anti- -NOR90, anti-Th/To, anti-PM-Scl-100, anti-PM-Scl-75, anti-Ku, anti-Ro-52, anti-PDGFR. The presence of antibodies was assessed using a test - EUROLINE Systemic Sclerosis Profile. RESULTS: 82 patients (94%) had positive antinuclear antibodies; anti-topo I - 29 patients; anti-CENP-A - 20 and anti-CENP-B - 20; anti-RP11 - 9 and anti-RP155 - 7; anti-U3RNP - 1; anti-NOR90 - 6; anti-Th/ To - 3; anti-PM-Scl-100 - 7; anti-PM-Scl-75 - 11; anti-Ku - 5; anti-Ro-52 - 23 patients. We found significant differences in prevalence of anti-topo I: 25/35 vs. 4/52 p=0.0000; anti-CENP A: 0/35 vs. 20/52 p=0.0001; anti-CENP B: 0/35 vs. 20/52 p=0.0001 between dcSSc and lcSSc. CONCLUSIONS: Some antibodies in SSc, e.g. anti-topo I and anti-centromere, are useful in defining the clinical subset of disease and provide prognostic information. There are no significant differences in the prevalence of other autoantibodies associated with SSc between dcSSc and lcSSc patients.
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Anticuerpos Antinucleares/sangre , Autoanticuerpos/sangre , Autoantígenos/sangre , Proteínas Cromosómicas no Histona/sangre , Esclerodermia Sistémica/sangre , Esclerodermia Sistémica/diagnóstico , Pruebas Serológicas/métodos , Anciano , Biomarcadores/sangre , Proteína A Centromérica , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , ARN Polimerasa III/sangre , Esclerodermia Sistémica/inmunologíaAsunto(s)
Proteínas Cromosómicas no Histona/genética , Depresión Posparto/genética , Epigénesis Genética , Predisposición Genética a la Enfermedad/genética , Proteínas del Tejido Nervioso/genética , Biomarcadores/sangre , Homólogo de la Proteína Chromobox 5 , Proteínas Cromosómicas no Histona/sangre , Metilación de ADN/efectos de los fármacos , Depresión Posparto/sangre , Estradiol/farmacología , Femenino , Humanos , Proteínas del Tejido Nervioso/sangreRESUMEN
The structural maintenance of chromosome 1 (Smc1) protein is a member of the highly conserved cohesin complex and is involved in sister chromatid cohesion. In response to ionizing radiation, Smc1 is phosphorylated at two sites, Ser-957 and Ser-966, and these phosphorylation events are dependent on the ATM protein kinase. In this study, we describe the generation of two novel ELISAs for quantifying phospho-Smc1(Ser-957) and phospho-Smc1(Ser-966). Using these novel assays, we quantify the kinetic and biodosimetric responses of human cells of hematological origin, including immortalized cells, as well as both quiescent and cycling primary human PBMC. Additionally, we demonstrate a robust in vivo response for phospho-Smc1(Ser-957) and phospho-Smc1(Ser-966) in lymphocytes of human patients after therapeutic exposure to ionizing radiation, including total-body irradiation, partial-body irradiation, and internal exposure to (131)I. These assays are useful for quantifying the DNA damage response in experimental systems and potentially for the identification of individuals exposed to radiation after a radiological incident.
Asunto(s)
Análisis Químico de la Sangre/métodos , Proteínas de Ciclo Celular/sangre , Proteínas Cromosómicas no Histona/sangre , Exposición a Riesgos Ambientales/análisis , Ensayo de Inmunoadsorción Enzimática/métodos , Fosfoproteínas/sangre , Radiometría/métodos , Adulto , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/inmunología , Proteínas de la Ataxia Telangiectasia Mutada , Proteínas de Ciclo Celular/química , Proteínas de Ciclo Celular/inmunología , Proteínas Cromosómicas no Histona/química , Proteínas Cromosómicas no Histona/inmunología , Daño del ADN , Proteínas de Unión al ADN/deficiencia , Relación Dosis-Respuesta en la Radiación , Femenino , Humanos , Radioisótopos de Yodo/efectos adversos , Linfocitos/metabolismo , Linfocitos/efectos de la radiación , Masculino , Datos de Secuencia Molecular , Fragmentos de Péptidos/química , Fragmentos de Péptidos/inmunología , Fosfoproteínas/química , Fosfoproteínas/inmunología , Proteínas Serina-Treonina Quinasas/deficiencia , Conejos , Factores de Tiempo , Proteínas Supresoras de Tumor/deficiencia , Irradiación Corporal Total/efectos adversos , Adulto JovenAsunto(s)
Anticuerpos Antiidiotipos/sangre , Autoantígenos/sangre , Enfermedades Autoinmunes/sangre , Proteína B del Centrómero/sangre , Centrómero/inmunología , Proteínas Cromosómicas no Histona/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Proteína A Centromérica , Humanos , Sensibilidad y EspecificidadRESUMEN
Systemic sclerosis, scleroderma, is a disease characterized by widespread vascular injury and fibrosis of the skin and visceral organs. Circulating autoantibodies against several intracellular antigens are common in scleroderma patients. The specificities of such autoantibodies correlate with distinct clinical manifestations. However, till date there is no evidence that these autoantibodies, though helpful in diagnosis and prognosis, are linked to the pathogenesis of scleroderma nor that they may cause any feature of the disease. Recently, the discovery of novel agonistic autoantibodies targeting the PDGF receptor has provided important insight into the molecular pathogenesis of scleroderma and the intracellular mechanisms leading to fibrosis. Although their pathogenic role awaits validation in in vivo models, these antibodies represent the molecular link between the immune system and fibrosis.
Asunto(s)
Autoanticuerpos/sangre , Autoinmunidad , Proteínas Cromosómicas no Histona/inmunología , ADN-Topoisomerasas de Tipo I/inmunología , Receptores del Factor de Crecimiento Derivado de Plaquetas/inmunología , Esclerodermia Sistémica/inmunología , Animales , Autoanticuerpos/inmunología , Proteínas Cromosómicas no Histona/sangre , ADN-Topoisomerasas de Tipo I/sangre , Células Endoteliales/inmunología , Fibroblastos/inmunología , Humanos , Receptores del Factor de Crecimiento Derivado de Plaquetas/sangre , Esclerodermia Sistémica/sangre , Esclerodermia Sistémica/terapiaRESUMEN
Inorganic mercury (mercuric chloride--HgCl(2)) induces in mice an autoimmune syndrome (HgIA) with T cell-dependent polyclonal B cell activation and hypergammaglobulinemia, dose- and H-2-dependent production of autoantibodies targeting the 34 kDa nucleolar protein fibrillarin (AFA), and systemic immune-complex deposits. The organic mercury species methylmercury (MeHg) and ethylmercury (EtHg--in the form of thimerosal) induce AFA, while the other manifestations of HgIA seen after treatment with HgCl(2) are present to varying extent. Since these organic Hg species are converted to the autoimmunogen Hg(2+) in the body, their primary autoimmunogen potential is uncertain and the subject of this study. A moderate dose of HgCl(2) (8 mg/L drinking water--internal dose 148 micro gHg/kg body weight [bw]/day) caused the fastest AFA response, while the induction was delayed after higher (25 mg/L) and lower (1.5 and 3 mg/L) doses. The lowest dose of HgCl(2) inducing AFA was 1.5 mg/L drinking water which corresponded to a renal Hg(2+) concentration of 0.53 micro g/g. Using a dose of 8 mg HgCl(2)/L this threshold concentration was reached within 24 h, and a consistent AFA response developed after 8-10 days. The time lag for the immunological part of the reaction leading to a consistent AFA response was therefore 7-9 days. A dose of thimerosal close to the threshold dose for induction of AFA (2 mg/L drinking water--internal dose 118 micro gHg/kg bw per day), caused a renal Hg(2+) concentration of 1.8 micro g/g. The autoimmunogen effect of EtHg might therefore be entirely due to Hg(2+) formed from EtHg in the body. The effect of organic and inorganic Hg species on T-helper type 1 and type 2 cells during induction of AFA was assessed as the presence and titre of AFA of the IgG1 and IgG2a isotype, respectively. EtHg induced a persistent Th1-skewed response irrespectively of the dose and time used. A low daily dose of HgCl(2) (1.5-3 mg/L) caused a Th1-skewed AFA response, while a moderate dose (8 mg/L) after 2 weeks resulted in a balanced or even Th2-skewed response. Higher daily doses of HgCl(2) (25 mg/L) caused a balanced Th2-Th1 response already from onset. In conclusion, while metabolically formed Hg(2+) might be the main AFA-inducing factor also after treatment with EtHg, the quality of the Hg-induced AFA response is modified by the species of Hg as well as the dose.
Asunto(s)
Autoinmunidad/inmunología , Activación de Linfocitos/efectos de los fármacos , Cloruro de Mercurio/toxicidad , Linfocitos T Colaboradores-Inductores/efectos de los fármacos , Animales , Anticuerpos Antinucleares/inmunología , Proteínas Cromosómicas no Histona/sangre , Relación Dosis-Respuesta a Droga , Femenino , Técnica del Anticuerpo Fluorescente , Inmunoglobulina G/clasificación , Inmunoglobulina G/inmunología , Inmunoglobulina G/metabolismo , Riñón/química , Riñón/efectos de los fármacos , Activación de Linfocitos/inmunología , Cloruro de Mercurio/química , Cloruro de Mercurio/inmunología , Ratones , Proteínas Nucleares/sangre , Órbita/irrigación sanguínea , Linfocitos T Colaboradores-Inductores/inmunología , Células TH1/efectos de los fármacos , Células TH1/inmunología , Células Th2/efectos de los fármacos , Células Th2/inmunología , Timerosal/química , Timerosal/inmunología , Timerosal/toxicidad , Factores de TiempoRESUMEN
AIMS AND BACKGROUND: The aims of this study were to assess the clinical utility of circulating preoperative HER-2 extracellular domain p105 detected by enzyme immunoassay (ELISA), to compare the tissue expression of HER-2/neu determined by immunohistochemistry (IHC), to correlate prognostic factors including tumor size, nodal involvement, and hormone receptor status, and to analyze the prognostic significance of the marker in relation to clinical outcome as measured by disease-free and overall survival. METHODS: In this study, we enrolled 108 consecutive patients with breast carcinoma, and obtained serum samples and frozen tumor tissues. We compared them with 57 women with fibroadenoma and 63 healthy women as controls. RESULTS: Univariate ANOVA analysis showed no relationship between HER-2/neu in tissue and serum. Preoperative serum levels of p105 were significantly higher in breast cancer patients than in women with benign disease or healthy women. Concerning the correlation between p105, HER-2/neu tissue expression, and the other prognostic factors, a statistically significant correlation between high serum p105 levels and ER-negative status in breast cancer patients was found. Kaplan-Meier analysis confirmed that patients with positive HER-2/neu tissue expression had a significantly shorter survival than those with negative expression. Analysis with the Cox model demonstrated that tumor size was the only significant independent prognostic factor. CONCLUSIONS: This research failed to demonstrate a relationship between preoperative tissue overexpression and circulating HER-2/neu, suggesting that p105 does not represent a valid alternative to predict a worsened prognosis in breast cancer, but it could be a diagnostic marker to discriminate healthy subjects from breast cancer patients.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/metabolismo , Receptor ErbB-2/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Análisis de Varianza , Antígenos Nucleares/sangre , Biomarcadores de Tumor/sangre , Neoplasias de la Mama/sangre , Neoplasias de la Mama/patología , Neoplasias de la Mama/cirugía , Proteínas Cromosómicas no Histona/sangre , Supervivencia sin Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Fibroadenoma/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , Inmunohistoquímica , Metástasis Linfática , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Pronóstico , Modelos de Riesgos Proporcionales , Receptor ErbB-2/sangre , Receptores de Estrógenos/metabolismo , Receptores de Progesterona/metabolismo , Estudios Retrospectivos , Factores de Riesgo , Análisis de Supervivencia , Regulación hacia ArribaRESUMEN
The mechanisms underlying the antifertility effects of hyperprolactinemia have yet to be established in an appropriate experimental model. Hyperprolactinemia is a known side effect of fluphenazine, a broad spectrum, long-acting phenothiazine known to be dopamine type-D2 receptor antagonist. In our earlier study in adult male rats, we reported that fluphenazine at a dose of 3 mg/kg/day suppressed serum FSH but not testosterone (T) through increasing dopamine (DA) metabolism in the pituitary gland, within 60 days. Fluphenazine treatment affected sperm quality and male rats treated with fluphenazine sired fewer litters. The effects of fluphenazine-induced hyperprolactinemia on sperm quality appeared to be related to reduced FSH. We now report that FSH suppression enhanced the uptake of acridine orange (AO), a DNA intercalating, fluorescent dye by the fluphenazine-treated caput epididymal sperms with concomitant reduction in the uptake of thiol-specific monobromobimane (mBBr) fluorescent dye in vitro, suggesting greater accessibility of DNA intercalating dye to sperm chromatin and reduction in free sperm protein thiols. The concomitant increase in AO and decrease in mBBr fluorescence was suggestive of loose chromatin packaging in caput epididymal sperms after treatment with fluphenazine at 3 mg/kg/day for 60 days. The suppression in levels of protamine (P1) in caput epididymal sperms suggested that chromatin hypocompaction was due to reduced deposition of protamines in sperm chromatin. Reduction in testicular levels of cyclic adenosyl 3', 5' monophosphate response element modulator (CREMtau) and P1 further suggested that reduced deposition was indeed due to reduced synthesis. The concomitant reduction in testicular levels of transition protein 1 (TP1) and transition protein 2 (TP2) also suggested that hypoprotamination was due to reduced synthesis of these proteins crucial for facilitating P1 deposition. The effect appeared to have occurred at the level of translation of CREMtau, since its transcript levels were unaffected whereas those of TP1, TP2 and P1 and protamine were upregulated. The study led to the view that the effects of FSH suppression were manifest on the posttranscriptional modifications of CREMtau, as also on transcript repression of TP1, TP2, P1, which do the RNA- binding proteins bring about. Reduction in FSH did not decrease ABP expression in the testis, which has recently been implicated in the expression of transition protein 1 in vitro. However, a significant reduction was evident after fluphenazine treatment, in the immunoexpression of testicular cAMP, the mediator of FSH effects in the Sertoli cells and putative mediator of ABP effects in the spermatids. The study suggests that fluphenazine-induced hyperprolactinemia suppressed FSH and affected a putative cAMP-dependent mechanism underlying posttranscriptional modification of spermatidal genes involved in chromatin condensation, presumably by reducing the availability/secretion of ABP, a paracrine regulator of spermiogenesis in vitro.
Asunto(s)
Hiperprolactinemia/fisiopatología , Espermatogénesis/fisiología , Animales , Antipsicóticos , Western Blotting , Proteínas Cromosómicas no Histona/sangre , AMP Cíclico/metabolismo , Modulador del Elemento de Respuesta al AMP Cíclico , Proteínas de Unión al ADN/sangre , Flufenazina , Expresión Génica/efectos de los fármacos , Hiperprolactinemia/inducido químicamente , Masculino , Protaminas/sangre , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Cromatina Sexual/metabolismo , Espermatozoides/efectos de los fármacos , Espermatozoides/metabolismo , Compuestos de Sulfhidrilo/metabolismo , Testículo/efectos de los fármacos , Testículo/metabolismo , Factores de Transcripción/sangreRESUMEN
BACKGROUND: Anticentromere antibodies (ACA) are frequently observed in patients with Raynaud's phenomenon and in the CREST syndrome, a subclass of systemic sclerosis. Likewise, ACA are also found in other autoimmune and non-autoimmune diseases. The objective of the present study was to evaluate the clinical utility of the measurement of antibodies to the best characterized centromere antigen (CENP-B) protein by an enzyme-linked immunosorbent assay (ELISA) that uses human recombinant CENP-B antigen and compare it with indirect immunofluorescence assay (IFA) on HEp-2 cells. METHODS: We have analyzed 128 sera samples from patients with the following diseases: systemic lupus erythematosus (SLE, n = 53), mixed connective tissue disease (n = 1), primary Sjögren syndrome (n = 10), primary Raynaud's phenomenon (n = 10), primary systemic sclerosis (n = 7), polymyositis/dermatomyositis (n = 3), rheumatoid arthritis (n = 9), cutaneous lupus (n = 5), primary biliary cirrhosis (n = 9), chronic autoimmune hepatitis (n = 5) and ANA-positive non-autoimmune diseases (n = 16). RESULTS: The ELISA evaluated shows a good concordance with IFA, with the advantage of being an automatable quantitative technique. CONCLUSIONS: Measurement of anticentromere antibodies by this ELISA using human recombinant antigen is a useful alternative for the autoimmune laboratory checking for diseases associated with anticentromere antibodies.