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1.
PLoS One ; 14(12): e0226823, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31877171

RESUMO

Cataracts are focal to diffuse opacities of the eye lens causing impaired vision or complete blindness. For bilateral congenital cataracts in Red Holsteins a perfectly cosegregating mutation within the CPAMD8 gene (CPAMD8:g.5995966C>T) has been reported. We genotyped the CPAMD8:g.5995966C>T variant in Holstein calves affected by congenital bilateral congenital cataracts, their unaffected relatives and randomly selected herd mates. Ophthalmological examinations were performed in all affected individuals to confirm a congenital cataract. Whole genome sequencing was employed to screen variants in candidate genes for the Morgagnian cataract phenotype. In the present study, 3/35 cases were confirmed as homozygous mutated and 6/14 obligate carriers. Further 7/46 unaffected animals related with these cases were heterozygous mutated for the CPAMD8:g.5995966C>T variant. However 32 cases with a congenital cataract showed the wild type for the CPAMD8 variant. We did not identify variants in the candidate genes CPAMD8 and NID1 or in their close neighborhood as strongly associated with the congenital cataract phenotype in Holstein calves with the CPAMD8 wild type. In conclusion, the CPAMD8:g.5995966C>T variant is insufficient to explain the majority of Morgagnian congenital cataract phenotypes in Holsteins. It is very likely that congenital bilateral cataracts may be genetically heterogeneous and not yet known variants in genes other than CPAMD8 and NID1 are involved.


Assuntos
Catarata/veterinária , Doenças dos Bovinos/congênito , Doenças dos Bovinos/patologia , Bovinos/genética , Animais , Catarata/genética , Catarata/patologia , Doenças dos Bovinos/genética , Complemento C3/genética , Feminino , Masculino , Polimorfismo de Nucleotídeo Único , Inibidor da Tripsina Pancreática de Kazal/genética , alfa-Macroglobulinas/genética
2.
Can J Vet Res ; 83(4): 267-271, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31571726

RESUMO

The objective of this study was to assess whether concentrations of epidermal growth factor (EGF), fibronectin, and alpha(α)-2-macroglobulin in canine serum remain stable under different storage conditions. Serum was obtained from 10 adult dogs and stored for 7 d at room temperature (RT) and at 4°C and for 1, 3, and 6 mo at -20°C. Bacterial cultures of serum were carried out after 7 d at 4°C and at RT. For each dog and time point, EGF, fibronectin, and α-2-macroglobulin were quantified in duplicate by enzyme-linked immunosorbent assay (ELISA). Mean concentrations of each factor at each time point were used for statistical analysis. No bacterial growth was observed in any samples. Compared to baseline (232.24 ± 49.47 pg/mL), EGF concentration was significantly lower after 1 wk of storage at 4°C (135.39 ± 27.12 pg/mL, P = 0.006), but not at RT (315.85 ± 79 pg/mL, P = 0.6) or after 1, 3, or 6 mo of storage at -20°C (220.84 ± 41.07 pg/mL, P = 0.7; 220.98 ± 78.26 pg/mL, P = 0.8; 266.06 ± 20.39 pg/mL, P = 0.4, respectively). Compared to baseline, concentrations of fibronectin after 1 wk of storage at 4°C or at RT and 1, 3, or 6 mo of storage at -20°C were not statistically different. Compared to baseline (186.67 ± 45.20 mg/dL), the concentration of α-2-macroglobulin after 1 wk of storage at 4°C was significantly increased (244.61 ± 58.27 mg/dL, P = 0.002), but not at RT (177.09 ± 26.99 mg/dL, P = 0.2). The differences in concentration after 3 and 6 mo of storage at -20°C were significant compared to baseline (243.32 ± 42.64 mg/dL, P = 0.005 and 56.39 ± 21.78 mg/dL, P < 0.0001, respectively), but not after 1 mo of storage at -20°C (136.79 ± 25.61 mg/dL, P = 0.1). One week of storage at RT has little effect on the stability of EGF, fibronectin, and α-2-macroglobulin in canine serum. Measured factors remain stable for 3 mo of storage at -20°C.


Assuntos
Cães/sangue , Fator de Crescimento Epidérmico/sangue , Fibronectinas/sangue , alfa-Macroglobulinas/metabolismo , Animais , Fator de Crescimento Epidérmico/química , Fibronectinas/química , Manejo de Espécimes , Fatores de Tempo
3.
Neurology ; 93(9): e851-e863, 2019 08 27.
Artigo em Inglês | MEDLINE | ID: mdl-31366724

RESUMO

OBJECTIVE: To validate the Genot-PA score, a clinical-genetic logistic regression score that stratifies the thrombolytic therapy safety, in a new cohort of patients with stroke. METHODS: We enrolled 1,482 recombinant tissue plasminogen activator (rtPA)-treated patients with stroke in Spain and Finland from 2003 to 2016. Cohorts were analyzed on the basis of ethnicity and therapy: Spanish patients treated with IV rtPA within 4.5 hours of onset (cohort A and B) or rtPA in combination with mechanical thrombectomy within 6 hours of onset (cohort C) and Finnish participants treated with IV rtPA within 4.5 hours of onset (cohort D). The Genot-PA score was calculated, and hemorrhagic transformation (HT) and parenchymal hematoma (PH) risks were determined for each score stratum. RESULTS: Genot-PA score was tested in 1,324 (cohort A, n = 726; B, n = 334; C, n = 54; and D, n = 210) patients who had enough information to complete the score. Of these, 213 (16.1%) participants developed HT and 85 (6.4%) developed PH. In cohorts A, B, and D, HT occurrence was predicted by the score (p = 2.02 × 10-6, p = 0.023, p = 0.033); PH prediction was associated in cohorts A through C (p = 0.012, p = 0.034, p = 5.32 × 10-4). Increased frequency of PH events from the lowest to the highest risk group was found (cohort A 4%-15.7%, cohort B 1.5%-18.2%, cohort C 0%-100%). The best odds ratio for PH prediction in the highest-risk group was obtained in cohort A (odds ratio 5.16, 95% confidence interval 1.46-18.08, p = 0.009). CONCLUSION: The Genot-PA score predicts HT in patients with stroke treated with IV rtPA. Moreover, in an exploratory study, the score was associated with PH risk in mechanical thrombectomy-treated patients.


Assuntos
Hemorragia Cerebral/epidemiologia , Valor Preditivo dos Testes , Ativador de Plasminogênio Tecidual/efeitos adversos , Idoso , Idoso de 80 Anos ou mais , Hemorragia Cerebral/induzido quimicamente , Fator XII/genética , Feminino , Finlândia/epidemiologia , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Espanha/epidemiologia , Acidente Vascular Cerebral/tratamento farmacológico , Trombectomia/efeitos adversos , Trombectomia/estatística & dados numéricos , Terapia Trombolítica/efeitos adversos , Fatores de Tempo , Ativador de Plasminogênio Tecidual/uso terapêutico , alfa-Macroglobulinas/genética
4.
Orthop Surg ; 11(3): 481-486, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31243924

RESUMO

OBJECTIVE: The present study aimed to identify the relationship of α-2-macroglobulin and microvascular vessel pathology with steroid-induced femoral head necrosis in the Southeast Chinese population. METHODS: This study enrolled 40 patients diagnosed with steroid-induced necrosis of the femoral head. Patients had various stages of femoral head necrosis. The differential expression of serum proteins and mRNA from patients with steroid-induced necrosis of the femoral head (SINFH) and healthy volunteers was analyzed by western blot and quantitative polymerase chain reaction (QT-PCR). The pathological change in osteocyte necrosis was indicated by hematoxylin and eosin stain and immunohistochemistry. RESULTS: Hematoxylin and eosin stain showed histopathology changes in the necrotic area of patients with steroid-induced INFH: bone trabeculae were fewer and thinner, became broken, fragmented and structurally disordered; intraosseous adipose cells became enlarged; the arrangement of the osteoblasts became irregular; and vacant bone lacunae increased. QT-PCR showed significantly lower levels of α-2-macroglobulin in the serum of patients with SINFH than in controls (P < 0.05). Immunohistochemical staining and western blotting demonstrated that the expression of α-2-macroglobulin was significantly decreased in the necrotic area of SINFH patients (P < 0.05). CONCLUSION: The α-2-macroglobulin may be associated with the pathology of SINFH. The multiple pathological reactions occur in SINFH and α-2-macroglobulin may serve as a potential biomarker for the diagnosis of SINFH or a promising therapeutic target.


Assuntos
Corticosteroides/efeitos adversos , Anti-Inflamatórios/efeitos adversos , Necrose da Cabeça do Fêmur/induzido quimicamente , Microvasos/patologia , alfa-Macroglobulinas/metabolismo , Adulto , Biomarcadores/sangue , Western Blotting , Estudos de Casos e Controles , China , Feminino , Necrose da Cabeça do Fêmur/sangue , Necrose da Cabeça do Fêmur/diagnóstico , Necrose da Cabeça do Fêmur/patologia , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase
5.
Hum Mutat ; 40(8): 1156-1171, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31009165

RESUMO

A genetic basis for otitis media is established, however, the role of rare variants in disease etiology is largely unknown. Previously a duplication variant within A2ML1 was identified as a significant risk factor for otitis media in an indigenous Filipino population and in US children. In this report exome and Sanger sequencing was performed using DNA samples from the indigenous Filipino population, Filipino cochlear implantees, US probands, Finnish, and Pakistani families with otitis media. Sixteen novel, damaging A2ML1 variants identified in otitis media patients were rare or low-frequency in population-matched controls. In the indigenous population, both gingivitis and A2ML1 variants including the known duplication variant and the novel splice variant c.4061 + 1 G>C were independently associated with otitis media. Sequencing of salivary RNA samples from indigenous Filipinos demonstrated lower A2ML1 expression according to the carriage of A2ML1 variants. Sequencing of additional salivary RNA samples from US patients with otitis media revealed differentially expressed genes that are highly correlated with A2ML1 expression levels. In particular, RND3 is upregulated in both A2ML1 variant carriers and high-A2ML1 expressors. These findings support a role for A2ML1 in keratinocyte differentiation within the middle ear as part of otitis media pathology and the potential application of ROCK inhibition in otitis media.


Assuntos
Regulação para Baixo , Perfilação da Expressão Gênica/métodos , Mutação , Otite Média/genética , Análise de Sequência de DNA/métodos , alfa-Macroglobulinas/genética , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Finlândia , Regulação da Expressão Gênica , Predisposição Genética para Doença , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Paquistão , Linhagem , Filipinas , Análise de Sequência de RNA , Transdução de Sinais , Estados Unidos , Adulto Jovem
6.
Biomark Med ; 13(2): 105-121, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30767554

RESUMO

AIM: Pancreatic cancer is one of the worst malignant tumors in prognosis. Therefore, to reduce the mortality rate of pancreatic cancer, early diagnosis and prompt treatment are particularly important. RESULTS: We put forward a new feature-selection method that was used to find clinical markers for pancreatic cancer by combination of Support Vector Machine Recursive Feature Elimination (SVM-RFE) and Large Margin Distribution Machine Recursive Feature Elimination (LDM-RFE) algorithms. As a result, seven differentially expressed genes were predicted as specific biomarkers for pancreatic cancer because of their highest accuracy of classification on cancer and normal samples. CONCLUSION: Three (MMP7, FOS and A2M) out of the seven predicted gene markers were found to encode proteins secreted into urine, providing potential diagnostic evidences for pancreatic cancer.


Assuntos
Algoritmos , Biomarcadores Tumorais/genética , Perfilação da Expressão Gênica , Pâncreas/patologia , Neoplasias Pancreáticas/diagnóstico , Máquina de Vetores de Suporte , Biomarcadores Tumorais/urina , Estudos de Casos e Controles , Humanos , Metaloproteinase 7 da Matriz/genética , Metaloproteinase 7 da Matriz/urina , Pâncreas/metabolismo , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/urina , Prognóstico , Proteínas Proto-Oncogênicas c-fos/genética , Proteínas Proto-Oncogênicas c-fos/urina , Taxa de Sobrevida , alfa-Macroglobulinas/genética , alfa-Macroglobulinas/urina
7.
J Proteome Res ; 18(3): 1264-1277, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30525646

RESUMO

Lupus nephritis (LN) is a severe clinical manifestation of systemic lupus erythematosus (SLE) associated with significant morbidity and mortality. Assessment of severity and activity of renal involvement in SLE requires a kidney biopsy, an invasive procedure with limited prognostic value. Noninvasive biomarkers are needed to inform treatment decisions and to monitor disease activity. Proteinuria is associated with disease progression in LN; however, the composition of the LN urinary proteome remains incompletely characterized. To address this, we profiled LN urine samples using complementary mass spectrometry-based methods:  protein gel fractionation, chemical labeling using tandem mass tags, and data-independent acquisition. Combining results from these approaches yielded quantitative information on 2573 unique proteins in urine from LN patients. A multiple-reaction monitoring (MRM) method was established to confirm eight proteins in an independent cohort of LN patients, and seven proteins (transferrin, α-2-macroglobulin, haptoglobin, afamin, α-1-antitrypsin, vimentin, and ceruloplasmin) were confirmed to be elevated in LN urine compared to healthy controls. In this study, we demonstrate that deep mass spectrometry profiling of a small number of patient samples can identify high-quality biomarkers that replicate in an independent LN disease cohort. These biomarkers are being used to inform clinical biomarker strategies to support longitudinal and interventional studies focused on evaluating disease progression and treatment efficacy of novel LN therapeutics.


Assuntos
Biomarcadores/urina , Lúpus Eritematoso Sistêmico/urina , Nefrite Lúpica/urina , Proteoma/genética , Adolescente , Adulto , Idoso , Biópsia , Proteínas de Transporte/urina , Ceruloplasmina/urina , Feminino , Glicoproteínas/urina , Haptoglobinas/urina , Humanos , Rim/metabolismo , Rim/patologia , Lúpus Eritematoso Sistêmico/genética , Lúpus Eritematoso Sistêmico/patologia , Nefrite Lúpica/genética , Nefrite Lúpica/patologia , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Prognóstico , Albumina Sérica Humana/urina , Transferrina/urina , Vimentina/urina , Adulto Jovem , alfa 1-Antitripsina/urina , alfa-Macroglobulinas/urina
8.
Hum Genet ; 138(8-9): 1043-1049, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29556725

RESUMO

Primary congenital glaucoma is a trabecular meshwork dysgenesis with resultant increased intraocular pressure and ocular damage. CYP1B1 mutations remain the most common identifiable genetic cause. However, important questions about the penetrance of CYP1B1-related congenital glaucoma remain unanswered. Furthermore, mutations in other genes have been described although their exact contribution and potential genetic interaction, if any, with CYP1B1 mutations are not fully explored. In this study, we employed modern genomic approaches to re-examine CYP1B1-related congenital glaucoma. A cohort of 193 patients (136 families) diagnosed with congenital glaucoma. We identified biallelic CYP1B1 mutations in 80.8% (87.5 and 66.1% in familial and sporadic cases, respectively, p < 0.0086). The large family size of the study population allowed us to systematically examine penetrance of all identified alleles. With the exception of c.1103G>A (p.R368H), previously reported pathogenic mutations were highly penetrant (91.2%). We conclude from the very low penetrance and genetic epidemiological analyses that c.1103G>A (p.R368H) is unlikely to be a disease-causing recessive mutation in congenital glaucoma as previously reported. All cases that lacked biallelic CYP1B1 mutations underwent whole exome sequencing. No mutations in LTBP2, MYOC or TEK were encountered. On the other hand, mutations were identified in genes linked to other ophthalmic phenotypes, some inclusive of glaucoma, highlighting conditions that might phenotypically overlap with primary congenital glaucoma (SLC4A4, SLC4A11, CPAMD8, and KERA). We also encountered candidate causal variants in genes not previously linked to human diseases: BCO2, TULP2, and DGKQ. Our results both expand and refine the genetic spectrum of congenital glaucoma with important clinical implications.


Assuntos
Citocromo P-450 CYP1B1/genética , Glaucoma/genética , Alelos , Proteínas de Transporte de Ânions/genética , Estudos de Coortes , Proteínas do Citoesqueleto/genética , Análise Mutacional de DNA/métodos , Proteínas do Olho/genética , Feminino , Testes Genéticos/métodos , Glicoproteínas/genética , Humanos , Pressão Intraocular/genética , Proteínas de Ligação a TGF-beta Latente/genética , Masculino , Mutação/genética , Linhagem , Penetrância , Fenótipo , Receptor TIE-2/genética , alfa-Macroglobulinas/genética
10.
Protein J ; 38(1): 23-29, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30564965

RESUMO

Alpha-2-macroglobulin (α2M) is a molecule generally associated with inflammation, and chronic inflammation is associated with ageing and cancer. The degree of inflammation was recently proposed to be considered as a biomarker of biological ageing. In this study, glycans attached to α2M were analysed in a human population of different ages by lectin-based protein microarray. Higher reactivity of α2M with several lectins was detected in older individuals indicating an increased content of specific monosaccharides: α2,6 sialic acid, mannose and N-acetylglucosamine, and multiantennary complex type N-glycans. The increased glycosylation of α2M was accompanied by reduced binding of Zn ions and insulin-like growth factor-binding protein 2 (IGFBP-2). Glycosylation of α2M and its reactivity with IGFBP-2 is similarly affected by ageing and incidence of colon cancer, but the reactivity of α2M with Zn ions is differently affected, as the binding of Zn ions remains unaltered in patients with colon cancer compared to healthy middle-aged individuals. Thus, the binding of IGFBP-2 to α2M seems to be related to structural changes in the glycan moieties of α2M, whereas binding of Zn ions, most likely, is not.


Assuntos
Envelhecimento/metabolismo , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Polissacarídeos/metabolismo , Zinco/metabolismo , alfa-Macroglobulinas/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Glicosilação , Humanos , Masculino , Pessoa de Meia-Idade
11.
Mol Hum Reprod ; 25(3): 137-155, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30590815

RESUMO

Glucose regulated protein 78 (GRP78) is expressed on cell surface in exceptional conditions as seen in cancer cells and macrophages. We have reported its membrane localization in sperm. The functional significance of its surface localization in sperm is an enigma. Alpha-2-macroglobulin (α2M) reportedly binds surface GRP78, regulating macrophage motility. Additionally, seminal plasma α2M levels are low in cases of asthenozoospermia. We investigated the functional relevance of sperm surface GRP78 and α2M crosstalk using testicular sperm (Tsp; immature) and caudal sperm (Cdsp; mature) from adult Holtzman rats. α2M colocalized and interacted with GRP78 and was significantly higher in Cdsp. Cofilin pathway proteins were detected in Tsp and Cdsp, however the pathway was highly active in Cdsp. Tsp surface GRP78 tyrosine phosphorylation and [Ca2+]i levels increased significantly on exposure to activated α2M (α2M*). This binding activated Rac/Cdc42, and consequently PAK, leading to LIMK and cofilin phosphorylation and thus promoting actin reorganization. Cofilin translocation from the sperm tail to the head in the presence of α2M* possibly prevented F-actin depolymerization in the tail. Thus, profiles observed with Cdsp could be re-created upon exposure of Tsp with α2M*. We conclude that α2M secreted into seminiferous tubule fluid by Sertoli cells, may be activated by proteinases in the epididymis and may bind to sperm surface GRP78 during epididymal transit, thereby facilitating sperm motility via actin reorganization. As F-actin is required for maintaining structural integrity and hyperactivated motility in sperm, our finding has significant implications in light of our previous reports of reduced GRP78 phosphorylation and the actin-based motility pathway being significantly altered in asthenozoospermia.


Assuntos
Actinas/metabolismo , Epididimo/metabolismo , Proteínas de Choque Térmico/metabolismo , alfa 2-Macroglobulinas Associadas à Gravidez/metabolismo , Espermatozoides/metabolismo , alfa-Macroglobulinas/metabolismo , Astenozoospermia/metabolismo , Cálcio/metabolismo , Humanos , Masculino , Fosforilação
12.
J Pregnancy ; 2018: 8367571, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30405914

RESUMO

There is not a single or combined screening method for preterm birth with high sensitivity which will truly identify the women at risk for preterm birth while also with high specificity to prevent unnecessary interventions and high treatment costs. Measurement of cervical length is the most cost-effective method that is used in clinical practice. Bedside tests have also been developed for detecting markers like fetal fibronectin, insulin-like growth factor binding protein-1 (IGFBP-1), interleukin-6, and placental alpha-macroglobulin-1. Taking the maternal history, health condition, and sociodemographical factors into consideration is recommended. Ultrasound markers apart from cervical length measurements as uterocervical angle and placental strain ratio are studied. Investigations on metabolomics, proteomics, and microRNA profiling have brought a new aspect on this subject. Maybe in the future, with clear identification of women at true risk for preterm birth, development of more effective preventive strategies will not be unfeasible.


Assuntos
Nascimento Prematuro/diagnóstico , Biomarcadores/sangue , Medida do Comprimento Cervical , Feminino , Fibronectinas/sangue , Humanos , Proteína 1 de Ligação a Fator de Crescimento Semelhante à Insulina/sangue , Interleucina-6/sangue , Metabolômica , MicroRNAs , Valor Preditivo dos Testes , Gravidez , Nascimento Prematuro/diagnóstico por imagem , Nascimento Prematuro/prevenção & controle , Proteômica , Ultrassonografia , alfa-Macroglobulinas
13.
Int J Biol Macromol ; 120(Pt B): 2285-2292, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30149080

RESUMO

Bicalutamide (BCT), a drug used in the treatment of prostate cancer, antagonises the actions of androgens, at the receptor level, thereby inhibiting the growth of prostate tumours. Alpha-2-macroglobulin (α2M), a pan-proteinase inhibitor, inhibits proteinase, regardless of specificity and catalytic mechanism. α2M is deficient in patients of advanced prostate cancer with bone metastases. Our studies explored the interaction of BCT with α2M and analysed the BCT induced structural alteration to the α2M. The result suggests that BCT decreases the antiproteolytic potential and causes structural and functional change in human α2M. UV-visible absorption spectroscopy confirms the formation of α2M-BCT complex. Fluorescence analysis shows significant quenching in fluorescence intensity of α2M upon binding with BCT. Synchronous fluorescence result suggests the interaction of BCT with α2M changed the microenvironment around tyrosine residues. Secondary structure of α2M also undergoes a slight change upon complexation with the drug as evident by shift in negative ellipticity in far UV CD spectroscopy. FTIR results confirm the alteration in secondary structure of α2M upon drug interaction. Molecular docking studies show that BCT bind to a monomer of α2M primarily through hydrophobic force. Thermodynamics parameters were determined by isothermal titration calorimetry found that the binding was exothermic in nature.


Assuntos
Antagonistas de Androgênios/metabolismo , Anilidas/metabolismo , Simulação de Acoplamento Molecular , Nitrilos/metabolismo , alfa 2-Macroglobulinas Associadas à Gravidez/metabolismo , Compostos de Tosil/metabolismo , alfa-Macroglobulinas/metabolismo , Humanos , Inibidores de Proteases/metabolismo , Ligação Proteica , Conformação Proteica , alfa-Macroglobulinas/química
14.
Arch Argent Pediatr ; 116(4): 275-282, 2018 Aug 01.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-30016029

RESUMO

INTRODUCTION: Obesity is a worldwide public health problem and the most common non-communicable chronic disease. It is associated with an increase in inflammatory acute phase proteins and proinflammatory cytokines. OBJECTIVE: To assess the levels of acute phase proteins in obese children and adolescents with hepatic steatosis and metabolic syndrome. Methodology. Forty-five children with a body mass index ≥ 95th percentile aged 5.0-15.5 years were included. The following acute phase reactants were determined: C-reactive protein, haptoglobin, alpha-2-macroglobulin, and apolipoprotein A-1; besides, an ultrasound was done to assess hepatic steatosis. RESULTS: C-reactive protein levels increased in all patients. Patients with metabolic syndrome also had high levels of apolipoprotein A-1 and haptoglobin. Patients with hepatic steatosis had a significant increase in alpha-2-macroglobulin in addition to high C-reactive protein.


Assuntos
Fígado Gorduroso/epidemiologia , Mediadores da Inflamação/metabolismo , Síndrome Metabólica/epidemiologia , Obesidade Pediátrica/epidemiologia , Adolescente , Apolipoproteína A-I/metabolismo , Biomarcadores/metabolismo , Índice de Massa Corporal , Proteína C-Reativa/metabolismo , Criança , Pré-Escolar , Citocinas/metabolismo , Fígado Gorduroso/fisiopatologia , Feminino , Haptoglobinas/metabolismo , Humanos , Estudos Longitudinais , Masculino , Síndrome Metabólica/fisiopatologia , Obesidade Pediátrica/fisiopatologia , Estudos Prospectivos , alfa-Macroglobulinas/metabolismo
15.
Crit Care Med ; 46(7): 1106-1113, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29912095

RESUMO

OBJECTIVES: Sepsis is associated with high early and total in-hospital mortality. Despite recent revisions in the diagnostic criteria for sepsis that sought to improve predictive validity for mortality, it remains difficult to identify patients at greatest risk of death. We compared the utility of nine biomarkers to predict mortality in subjects with clinically suspected bacterial sepsis. DESIGN: Cohort study. SETTING: The medical and surgical ICUs at an academic medical center. SUBJECTS: We enrolled 139 subjects who met two or more systemic inflammatory response syndrome (systemic inflammatory response syndrome) criteria and received new broad-spectrum antibacterial therapy. INTERVENTIONS: We assayed nine biomarkers (α-2 macroglobulin, C-reactive protein, ferritin, fibrinogen, haptoglobin, procalcitonin, serum amyloid A, serum amyloid P, and tissue plasminogen activator) at onset of suspected sepsis and 24, 48, and 72 hours thereafter. We compared biomarkers between groups based on both 14-day and total in-hospital mortality and evaluated the predictive validity of single and paired biomarkers via area under the receiver operating characteristic curve. MEASUREMENTS AND MAIN RESULTS: Fourteen-day mortality was 12.9%, and total in-hospital mortality was 29.5%. Serum amyloid P was significantly lower (4/4 timepoints) and tissue plasminogen activator significantly higher (3/4 timepoints) in the 14-day mortality group, and the same pattern held for total in-hospital mortality (Wilcoxon p ≤ 0.046 for all timepoints). Serum amyloid P and tissue plasminogen activator demonstrated the best individual predictive performance for mortality, and combinations of biomarkers including serum amyloid P and tissue plasminogen activator achieved greater predictive performance (area under the receiver operating characteristic curve > 0.76 for 14-d and 0.74 for total mortality). CONCLUSIONS: Combined biomarkers predict risk for 14-day and total mortality among subjects with suspected sepsis. Serum amyloid P and tissue plasminogen activator demonstrated the best discriminatory ability in this cohort.


Assuntos
Estado Terminal/mortalidade , Sepse/mortalidade , Idoso , Biomarcadores/sangue , Proteína C-Reativa/análise , Estudos de Coortes , Ferritinas/sangue , Fibrinogênio/análise , Haptoglobinas/análise , Mortalidade Hospitalar , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Pró-Calcitonina/sangue , Sepse/sangue , Sepse/diagnóstico , Proteína Amiloide A Sérica/análise , Componente Amiloide P Sérico/análise , Ativador de Plasminogênio Tecidual/sangue , alfa-Macroglobulinas/análise
16.
PLoS One ; 13(4): e0195610, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29684087

RESUMO

BACKGROUND: We previously identified ovostatin 2 (OVOS2) as a new candidate gene for cutaneous malignant melanoma (CMM) in a Chinese population. In this study, we aimed to investigate the exact role of OVOS2 in cell proliferation, invasion, and tumorigenesis of melanoma A375 cells. METHODS: The downregulation of OVOS2 expression was performed using lentiviral vectors with specific shRNA. The effects of OVOS2 expression on cell proliferation, cell cycle, cell migration, cell invasion, and potential of tumorigenesis were further investigated. RESULTS: The downregulation of OVOS2 significantly suppressed the proliferation of A375 cells and led to a G2/M phase block. The transwell cell migration assay showed that the reduced expression of OVOS2 also significantly inhibited the transmigration of A375 cells. The western blot results showed downregulated expression of p-FAK, p-AKT, and p-ERK. This was accompanied by the upregulated epithelial phenotypes E-cadherin and ß-catenin, and downregulated expression of mesenchymal phenotype N-cadherin after OVOS2 knockdown. The transplantation tumor experiment in BALB/C nude mouse showed that after an observation period of 32 days, the growth speed and weight of the transplanted tumors were significantly suppressed in the BALB/c nude mice subcutaneously injected with OVOS2 knocked-down A375 cells. CONCLUSION: The inhibition of OVOS2 had significant suppressive effects on the proliferation, motility, and migration capabilities of A375 cells, suggesting a crucial promotive role of OVOS2 in the pathogenesis and progression of CMM. The involved mechanisms are at least partly associated with the overactivation of FAK/MAPK/ERK and FAK/PI3K/AKT signals.


Assuntos
Carcinogênese/metabolismo , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Melanoma/metabolismo , Invasividade Neoplásica/fisiopatologia , Neoplasias Cutâneas/metabolismo , alfa-Macroglobulinas/metabolismo , Animais , Apoptose/fisiologia , Carcinogênese/patologia , Ciclo Celular/fisiologia , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Melanoma/patologia , Camundongos Endogâmicos BALB C , Camundongos Nus , Invasividade Neoplásica/patologia , Transplante de Neoplasias , RNA Mensageiro/metabolismo , Distribuição Aleatória , Neoplasias Cutâneas/patologia , alfa-Macroglobulinas/antagonistas & inibidores , alfa-Macroglobulinas/genética
17.
Prostate ; 78(11): 819-829, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29659051

RESUMO

BACKGROUND: Prostate cancer cells produce high levels of the serine protease Prostate-Specific Antigen (PSA). PSA is enzymatically active in the tumor microenvironment but is presumed to be enzymatically inactive in the blood due to complex formation with serum protease inhibitors α-1-antichymotrypsin and α-2-macroglobulin (A2M). PSA-A2M complexes cannot be measured by standard ELISA assays and are also rapidly cleared from the circulation. Thus the exact magnitude of PSA production by prostate cancer cells is not easily measured. The PSA complexed to A2M is unable to cleave proteins but maintains the ability to cleave small peptide substrates. Thus, in advanced prostate cancer, sufficient PSA-A2M may be in circulation to effect total A2M levels, levels of cytokines bound to A2M and hydrolyze small circulating peptide hormones. METHODS: Total A2M levels in men with advanced prostate cancer and PSA levels above 1000 ng/mL were measured by ELISA and compared to controls. Additional ELISA assays were used to measure levels of IL-6 and TGF-beta which can bind to A2M. The ability of PSA-A2M complexes to hydrolyze protein and peptide substrates was analyzed ± PSA inhibitor. Enzymatic activity of PSA-A2M in serum of men with high PSA levels was also assayed. RESULTS: Serum A2M levels are inversely correlated with PSA levels in men with advanced prostate cancer. Il-6 Levels are significantly elevated in men with PSA >1000 ng/mL compared to controls with PSA <0.1 ng/mL. PSA-A2M complex in serum of men with PSA levels >1000 ng/mL can hydrolyze small fluorescently labeled peptide substrates but not large proteins that are PSA substrates. PSA can hydrolyze small peptide hormones like PTHrP and osteocalcin. PSA complexed to A2M retains the ability to degrade PTHrP. CONCLUSIONS: In advanced prostate cancer with PSA levels >1000 ng/mL, sufficient PSA-A2M is present in circulation to produce enzymatic activity against circulating small peptide hormones. Sufficient PSA is produced in advanced prostate cancer to alter total A2M levels, which can potentially alter levels of a variety of growth factors such as IL-6, TGF-beta, basic FGF, and PDGF. Alterations in levels of these cytokines and proteolytic degradation of small peptide hormones may have profound effect on host-cancer interaction.


Assuntos
Calicreínas/sangue , Osteocalcina/sangue , Proteína Relacionada ao Hormônio Paratireóideo/sangue , Antígeno Prostático Específico/sangue , Neoplasias da Próstata/sangue , alfa-Macroglobulinas/metabolismo , Ácidos Borônicos/farmacologia , Estudos de Casos e Controles , Feminino , Humanos , Calicreínas/antagonistas & inibidores , Masculino , Peptidomiméticos/farmacologia , Antígeno Prostático Específico/antagonistas & inibidores , Neoplasias da Próstata/patologia , alfa-Macroglobulinas/antagonistas & inibidores
18.
Sci Rep ; 8(1): 2421, 2018 02 05.
Artigo em Inglês | MEDLINE | ID: mdl-29402968

RESUMO

RASopathies are a group of heterogeneous conditions caused by germline mutations in RAS/MAPK signalling pathway genes. With next-generation sequencing (NGS), sequencing capacity is no longer a limitation to molecular diagnosis. Instead, the rising number of variants of unknown significance (VUSs) poses challenges to clinical interpretation and genetic counselling. We investigated the potential of an integrated pipeline combining NGS and the functional assessment of variants for the diagnosis of RASopathies. We included 63 Chinese patients with RASopathies that had previously tested negative for PTPN11 and HRAS mutations. In these patients, we performed a genetic analysis of genes associated with RASopathies using a multigene NGS panel and Sanger sequencing. For the VUSs, we evaluated evidence from genetic, bioinformatic and functional data. Twenty disease-causing mutations were identified in the 63 patients, providing a primary diagnostic yield of 31.7%. Four VUSs were identified in five patients. The functional assessment supported the pathogenicity of the RAF1 and RIT1 VUSs, while the significance of two VUSs in A2ML1 remained unclear. In summary, functional analysis improved the diagnostic yield from 31.7% to 36.5%. Although technically demanding and time-consuming, a functional genetic diagnostic analysis can ease the clinical translation of these findings to aid bedside interpretation.


Assuntos
Síndrome de Costello/genética , Displasia Ectodérmica/genética , Insuficiência de Crescimento/genética , Cardiopatias Congênitas/genética , Neurofibromatose 1/genética , Síndrome de Noonan/genética , Proteínas Proto-Oncogênicas c-raf/genética , Proteínas ras/genética , Adolescente , Animais , Bioensaio , Criança , Pré-Escolar , Biologia Computacional , Síndrome de Costello/patologia , Displasia Ectodérmica/patologia , Facies , Insuficiência de Crescimento/patologia , Feminino , Expressão Gênica , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Mutação em Linhagem Germinativa , Cardiopatias Congênitas/patologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Lactente , MAP Quinase Quinase 1/genética , Masculino , Mutação de Sentido Incorreto , Neurofibromatose 1/patologia , Síndrome de Noonan/patologia , Proteínas Proto-Oncogênicas p21(ras)/genética , Proteína SOS1/genética , Peixe-Zebra , alfa-Macroglobulinas/genética
19.
Int J Mol Sci ; 19(1)2018 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-29301248

RESUMO

Thyroid hormone is a potent stimulator of metabolism, playing a critical role in regulating energy expenditure and in key physiological mechanisms, such as growth and development. Although administration of thyroid hormone in the form of levo thyroxine (l-thyroxine) has been used to treat hypothyroidism for many years, the precise molecular basis of its physiological actions remains uncertain. Our objective was to define the changes in circulating protein levels that characterize alterations in thyroid hormone status. To do this, an integrated untargeted proteomic approach with network analysis was used. This study included 10 age-matched subjects with newly diagnosed overt hypothyroidism. Blood was collected from subjects at baseline and at intervals post-treatment with l-thyroxine until they reached to euthyroid levels. Plasma protein levels were compared by two-dimensional difference in gel electrophoresis (2D-DIGE) pre- and post-treatment. Twenty differentially expressed protein spots were detected. Thirteen were identified, and were found to be unique protein sequences by MALDI-TOF mass spectrometry. Ten proteins were more abundant in the hypothyroid vs. euthyroid state: complement C2, serotransferrin, complement C3, Ig κ chain C region, α-1-antichymotrypsin, complement C4-A, haptoglobin, fibrinogen α chain, apolipoprotein A-I, and Ig α-1 chain C region. Three proteins were decreased in abundance in the hypothyroid vs. euthyroid state: complement factor H, paraneoplastic antigen-like protein 6A, and α-2-macroglobulin. The differentially abundant proteins were investigated by Ingenuity Pathway Analysis (IPA) to reveal their associations with known biological functions. Their connectivity map included interleukin-6 (IL-6) and tumour necrosis factor α (TNF-α) as central nodes and the pathway identified with the highest score was involved in neurological disease, psychological disorders, and cellular movement. The comparison of the plasma proteome between the hypothyroid vs euthyroid states revealed differences in the abundance of proteins involved in regulating the acute phase response.


Assuntos
Terapia de Reposição Hormonal/efeitos adversos , Hipotireoidismo/sangue , Proteoma/metabolismo , Adulto , Apolipoproteínas/sangue , Complemento C3/metabolismo , Fator H do Complemento/metabolismo , Feminino , Fibrinogênio/metabolismo , Redes Reguladoras de Genes , Humanos , Hipotireoidismo/terapia , Interleucina-6/sangue , Masculino , Pessoa de Meia-Idade , Mapas de Interação de Proteínas , Transferrina/metabolismo , Fator de Necrose Tumoral alfa/sangue , alfa-Macroglobulinas/metabolismo
20.
Mol Psychiatry ; 23(11): 2192-2208, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-29302075

RESUMO

To date, gene-environment (GxE) interaction studies in depression have been limited to hypothesis-based candidate genes, since genome-wide (GWAS)-based GxE interaction studies would require enormous datasets with genetics, environmental, and clinical variables. We used a novel, cross-species and cross-tissues "omics" approach to identify genes predicting depression in response to stress in GxE interactions. We integrated the transcriptome and miRNome profiles from the hippocampus of adult rats exposed to prenatal stress (PNS) with transcriptome data obtained from blood mRNA of adult humans exposed to early life trauma, using a stringent statistical analyses pathway. Network analysis of the integrated gene lists identified the Forkhead box protein O1 (FoxO1), Alpha-2-Macroglobulin (A2M), and Transforming Growth Factor Beta 1 (TGF-ß1) as candidates to be tested for GxE interactions, in two GWAS samples of adults either with a range of childhood traumatic experiences (Grady Study Project, Atlanta, USA) or with separation from parents in childhood only (Helsinki Birth Cohort Study, Finland). After correction for multiple testing, a meta-analysis across both samples confirmed six FoxO1 SNPs showing significant GxE interactions with early life emotional stress in predicting depressive symptoms. Moreover, in vitro experiments in a human hippocampal progenitor cell line confirmed a functional role of FoxO1 in stress responsivity. In secondary analyses, A2M and TGF-ß1 showed significant GxE interactions with emotional, physical, and sexual abuse in the Grady Study. We therefore provide a successful 'hypothesis-free' approach for the identification and prioritization of candidate genes for GxE interaction studies that can be investigated in GWAS datasets.


Assuntos
Depressão/genética , Transtorno Depressivo/genética , Testes Genéticos/métodos , Adulto , Animais , Estudos de Coortes , Depressão/metabolismo , Modelos Animais de Doenças , Feminino , Proteína Forkhead Box O1/genética , Proteína Forkhead Box O1/metabolismo , Interação Gene-Ambiente , Predisposição Genética para Doença/genética , Estudo de Associação Genômica Ampla/métodos , Genótipo , Humanos , Masculino , MicroRNAs/análise , MicroRNAs/genética , Pessoa de Meia-Idade , Proteínas do Tecido Nervoso/genética , Polimorfismo de Nucleotídeo Único/genética , Gravidez , Ratos , Transcriptoma/genética , Fator de Crescimento Transformador beta1/genética , alfa-Macroglobulinas/genética , alfa-Macroglobulinas/metabolismo
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