RESUMO
The lymphopenia exhibited in patients with COVID-19 has been associated with a worse prognosis in the development of the disease. To understand the factors associated with a worse evolution of COVID-19, we analyzed comorbidities, indicators of inflammation such as CRP and the ratio of neutrophils/lymphocytes, as well as the count of blood cells with T-lymphocyte subtypes in 172 hospitalized patients with COVID-19 pneumonia. Patients were grouped according to their needs for mechanical ventilation (ICU care) or not. Within the comorbidities studied, obesity was the only associated with greater severity and ICU admission. Both the percentage and the absolute number of neutrophils were higher in patients needing ICU care than non-ICU patients, whereas absolute lymphocyte count, and especially the percentage of lymphocytes, presented a deep decline in critical patients. There was no difference between the two groups of patients for CD4 T-lymphocytes, neither in percentage of lymphocyte nor in absolute number, however for CD8 T-cells the differences were significant for both parameters which were in decline in ICU patients. There was a firm correlation between the highest values of inflammation indicators with the decrease in percentage of CD8 T-lymphocytes. This effect was not seen with CD4 cells. Obesity together with lymphopenia, especially whether preferentially affects to CD8 T- lymphocytes, are factors that can predict a poor prognosis in patients with COVID-19.
Assuntos
Betacoronavirus/patogenicidade , Linfócitos T CD8-Positivos/patologia , Infecções por Coronavirus/imunologia , Linfopenia/imunologia , Neutrófilos/patologia , Obesidade/imunologia , Pneumonia Viral/imunologia , Idoso , Idoso de 80 Anos ou mais , Betacoronavirus/imunologia , Biomarcadores/sangue , Proteína C-Reativa/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/patologia , Linfócitos T CD4-Positivos/virologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/virologia , COVID-19 , Estudos de Casos e Controles , Infecções por Coronavirus/complicações , Infecções por Coronavirus/mortalidade , Infecções por Coronavirus/terapia , Feminino , Humanos , Unidades de Terapia Intensiva , Contagem de Linfócitos , Linfopenia/complicações , Linfopenia/mortalidade , Linfopenia/terapia , Masculino , Pessoa de Meia-Idade , Neutrófilos/imunologia , Neutrófilos/virologia , Obesidade/complicações , Obesidade/mortalidade , Obesidade/terapia , Pandemias , Pneumonia Viral/complicações , Pneumonia Viral/mortalidade , Pneumonia Viral/terapia , Prognóstico , Respiração Artificial , Estudos Retrospectivos , SARS-CoV-2 , Índice de Gravidade de Doença , Análise de SobrevidaRESUMO
Background Breast cancer is the most common malignant tumor in women in the world. In 2005, it was incorporated to the Explicit Guaranties Health System (GES) in Chile. Aim To describe the demographic and clinical characteristics of breast cancer patients and to determine the effect of incorporating these women to GES. Material and Methods Medical records of 5,119 women with breast cancer aged 59 ± 14 years, attended at six public hospitals between 2000 and 2010 were reviewed. Median follow up was 87 months (range 1-182). Mortality was assessed using death certificates obtained at the National Identification Registry. Results Sixty six percent of women were in stage I-II, 29% in stage III and 5% in stage IV. Surgery was performed in 4023/5119 cases (79%), adyuvant radiotherapy in 3627/4517 cases (80%), chemotherapy in 3,204/3,424 cases (94%) and hormone therapy in 1,695/2,375 cases (71%). Between 2000 and 2010, there was a significant increase in the proportion of cases in stage I, from 8% to 25%, (p < 0.01). Overall survival (OS) increased 1% per year, since the beginning of GES system (p = 0.024). Five year OS was 75.1%. The figures for Stage I, II, III and IV were 93, 84, 62 and 27% respectively (p < 0.01). Patients without lymph node involvement and who were not triple negative, had a significantly better OS. Conclusions There was a significant increase in stage I cases, and a 1% per year OS improvement after GES system started, compared with the previous period.
Assuntos
Neoplasias da Mama/mortalidade , Neoplasias da Mama/terapia , Carcinoma/mortalidade , Carcinoma/terapia , Programas Nacionais de Saúde/estatística & dados numéricos , Garantia da Qualidade dos Cuidados de Saúde/estatística & dados numéricos , Adolescente , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/diagnóstico , Carcinoma/diagnóstico , Chile/epidemiologia , Feminino , Humanos , Estimativa de Kaplan-Meier , Masculino , Prontuários Médicos , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
Nucleolar staining of antinuclear antibodies (ANAs) is not exclusive to patients suffering systemic sclerosis (SSc) since it can occur in other autoimmune diseases, such as systemic lupus erythematosus (SLE). The nucleolar ANA pattern presents a low incidence in patients with SLE, with less than 9% reported in some studies. The significance of nucleolar staining and antinucleolar antibodies (ANoA) in SLE is still unknown, as is its association with clinical manifestations. To address these issues, a case-control study was carried out. Twenty-eight cases of SLE with nucleolar staining were enrolled, as well as 73 controls with no nucleolar staining and different ANA patterns (homogeneous, speckled, and combined homogeneous and speckled). The homogeneous nucleolar pattern was the most frequent (27 out of 28), and in 75% was combined with other ANA patterns. The anti-double stranded DNA antibodies showed no differences between the two groups of patients, nor the auto-antibodies detected by line immunoassay (LIA). However, we have found an increased frequency of anti-PM-Scl antibodies with respect to the controls (p = 0.02), in addition to the association between Raynaud's phenomenon (RP) and anti-PM-Scl antibodies (OR = 20.72, 95% CI 1.33-323.19, p = 0.03). Moreover, the cases of SLE showed a 7.78-fold increase in the risk of developing cancer (95%, CI 1.85-32.75, p = 0.005) with respect to the control group. Taken together these findings suggest that nucleolar staining represents a comorbidity factor in patients with SLE, although its significance must still be determined.
Assuntos
Anticorpos Antinucleares/metabolismo , Autoanticorpos/metabolismo , Nucléolo Celular/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Neoplasias/diagnóstico , Adulto , Anticorpos Antinucleares/sangue , Autoanticorpos/sangue , Estudos de Casos e Controles , Comorbidade , Feminino , Células Hep G2 , Humanos , Lúpus Eritematoso Sistêmico/sangue , Masculino , Pessoa de Meia-Idade , Neoplasias/sangue , Neoplasias/imunologia , Fatores de Risco , EspanhaRESUMO
BACKGROUND: Allergen-specific immunotherapy (SIT) is the only intervention for IgE-mediated respiratory disorders. OBJECTIVE: The aim of the study was to investigate the immunological modifications induced by SIT in patients allergic to olive and/or grass pollen by attempting to establish an association between these modifications and clinical improvements. METHODS: We studied 29 patients who were allergic to olive and/or grass pollen. Patients were randomized to 2 groups: an active treatment group, comprising 19 allergic patients who received SIT, and a control group, formed by 10 allergic patients who received pharmacological treatment for their allergic symptoms but not immunotherapy. We used flow cytometry to analyze intracellular expression of the cytokines IL-4, IFN-gamma, IL-10, and TGF-beta1 in CD4+ T cells, as well as expression of Foxp3, the costimulatory CTLA-4 molecule, and the non-costimulatory CD40L molecule. To assess clinical changes, patients recorded their medication consumption, symptoms, and the limitation of daily activities using diary cards and quality of life questionnaires. RESULTS: Six months after initiation of SIT, we recorded a reduction in cell surface CD40L expression in the CD4+ T-cell population and a shift in the cytokine production profile (decrease in IL-4-producing CD4+ T cells and increase in IFN-gamma, IL-10, and TGF-beta1). These changes persisted after 12 months. Simultaneously, a clinical improvement was observed. CONCLUSIONS: SIT-induced clinical improvement is the result of immunological modifications such as a reduction in CD40L expression on CD4 cells and alteration in the cytokine production profile.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Ligante de CD40/análise , Citocinas/biossíntese , Dessensibilização Imunológica , Rinite Alérgica Sazonal/terapia , Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Rinite Alérgica Sazonal/imunologia , Inquéritos e QuestionáriosRESUMO
Systemic lupus erythematosus (SLE) is an autoimmune systemic disease caused as a result of an imbalance of Th1-/Th2-type cytokines. The soluble form of CD30 (CD30s) released from peripheral blood cells has been described as a marker of active disease in Th2-type immune response as in SLE. However, the expression of CD30 on CD3 T lymphocytes from patients with SLE has not been studied yet. Therefore, we have addressed our study to attempt this issue, studying CD30 expression by flow cytometry on CD3 T lymphocytes and CD4/CD8 subsets in samples from SLE patients mainly with lupus nephritis. In parallel, we have determined the production of the cytokines IL-4 (Th2), IFNγ (Th1), IL-10 and TGFß by intracellular staining. Differences between positive CD30 T cells in healthy controls and patients with SLE were found, with a higher percentage of CD30-expressing T cells in patients with SLE (P = 0.001). In contrast to healthy controls, CD30 was mainly expressed on CD8 T cells from patients with SLE. The intracellular cytokine staining showed that TGFß is the main cytokine expressed in CD3 T cells from patients with SLE. In addition to this, we have found a positive correlation between CD30-expressing T cells and IL-4, IFNγ, and immunosuppressive cytokines (IL-10 and TGFß) (P < 0.05). These results suggest that CD30 could play a role in the pathogenesis of SLE and its expression on CD3 T lymphocytes is not restricted only to Th2-type response.
Assuntos
Complexo CD3/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Antígeno Ki-1/biossíntese , Lúpus Eritematoso Sistêmico/imunologia , Subpopulações de Linfócitos T/imunologia , Adulto , Biomarcadores/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Feminino , Humanos , Interferon gama/metabolismo , Interleucina-10/metabolismo , Interleucina-4/metabolismo , Interleucina-8/metabolismo , Lúpus Eritematoso Sistêmico/metabolismo , Nefrite Lúpica/imunologia , Nefrite Lúpica/metabolismo , Masculino , Subpopulações de Linfócitos T/metabolismo , Fator de Crescimento Transformador beta/metabolismoRESUMO
Most patients with Hymenoptera venom allergy (HVA) to vespid venoms present double sensitization by specific IgE (sIgE)-mediated cross-reactivity. Thus, it is mandatory could discriminate between a true double and primary sensitization to implement an accurate venom-specific immunotherapy (VIT). To date, CAP-inhibition is the reference method in the diagnosis of cross-reactivity in double sensitized patients to vespid venoms, being the results obtained with the component resolved diagnostics (CRD) conflicting. For this, we have studied in a cohort of double sensitized patients to Vespula vulgaris (VV) and Polistes dominulus (PD) venoms (n = 40) the diagnostic accuracy of CRD using the CAP-inhibition as reference method, as well as to investigate whether basophil activation test (BAT) is an alternative method for inconclusive results obtained by CAP-inhibition. CAP-inhibition showed a sensitivity of 59.46 % in view of the indeterminate results; most patients had true double sensitization (54.5 %), followed by single sensitization to PD (27.27 %) and VV (18.18 %) venoms. CRD based on rVes v 5/rPol d 5 (or vice versa) ratio as well as whole extracts I3/I77 (or vice versa) ratio (specific IgE-I3 to VV/specific IgE-I77 to PD) showed a low diagnostic accuracy (AUC = 0.504, p = 0.974; AUC = 0.35, p = 0.235; respectively). BAT was determined in parallel with CAP-inhibition in 12 patients, presented higher sensitivity than CAP-inhibition (p = 0.021) and a positive agreement of 71.43 %. Likewise it was able to identify 100% of inconclusive results, showing a specificity of 83.3 %. Therefore, CRD is not a suitable method to distinguish monosensitization and BAT appears to be an appropriate method resolving indeterminate results from the gold standard method.
Assuntos
Venenos de Abelha , Hipersensibilidade , Mordeduras e Picadas de Insetos , Alérgenos , Teste de Degranulação de Basófilos , Dessensibilização Imunológica , Humanos , Hipersensibilidade/diagnóstico , Imunoglobulina E , Venenos de VespasRESUMO
Human embryonic stem cells (hESCs) have an unlimited capacity to proliferate by a self-renewal process and can be differentiated in the three germ layers, opening doors to new clinical therapies to replace missing or damaged cells. The number of research groups and projects using human stem cells has increased largely in the last 5 yr. The creation of stem cell banks is another important step to support the advance of research in this field. Banks must be operated within the strict regulatory famework of good manufacturing practices and good laboratory practices that assure the highest quality standards and must implement a quality system that complies with international quality systems standards. It may also be appropriate to aim at an accreditation in order to assure correct laboratory practices at all times. Stem cell banks should receive the lines previously derived by other groups and hESCs should be provided for groups that justify their use in a research project previously approved by an ethical committee. The assays generally accepted as typical of hESCs together with the microbiological analysis should be performed in order to assure a consistent, reliable, and safe line for the researchers. In this article, the Andalusian Stem Cell Bank proposes a model of a stem cell banking process in order to create a flow diagram of hESC lines and, following the international initiatives in stem cells research, to achieve the full characterization of cells and a standardization of protocols that would simplify the hESCs culture.
Assuntos
Células-Tronco Embrionárias/citologia , Bancos de Tecidos , Linhagem Celular , Bases de Dados de Ácidos Nucleicos , Perfilação da Expressão Gênica , HumanosRESUMO
Colon cancer is a common disease that can be sporadic or familial. An inactivated adenomatous polyposis coli (APC) suppressor gene is found in over 80% of colorectal tumors, this being an early alteration in the development of adenomatous polyps. APC function is not only critical for tumor initiation and progression, and chromosome instability (CIN) is another characteristic dependent at least partly on APC mutations.
Assuntos
Instabilidade Cromossômica , Neoplasias Colorretais/genética , Genes APC , HumanosRESUMO
During the assembly of the HLA class I molecules with peptides in the peptide-loading complex, a series of transient interactions are made with ER-resident chaperones. These interactions culminate in the trafficking of the HLA class I molecules to the cell surface and presentation of peptides to CD8(+) T lymphocytes. Within the peptide-loading complex, the glycoprotein tapasin exhibits a relevant function. This immunoglobulin (Ig) superfamily member in the endoplasmic reticulum membrane tethers empty HLA class I molecules to the transporter associated with antigen-processing (TAP) proteins. This review will address the current concepts regarding the double role that tapasin plays in the peptide optimization and surface expression of the HLA class I molecules.
Assuntos
Retículo Endoplasmático/metabolismo , Antígenos de Histocompatibilidade Classe I/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Chaperonas Moleculares/metabolismo , Peptídeos/metabolismo , Animais , Apresentação de Antígeno , Linfócitos T CD8-Positivos/metabolismo , Complexo de Golgi/metabolismo , Antígenos de Histocompatibilidade Classe I/genética , Humanos , Proteínas de Membrana Transportadoras/genética , Camundongos , Chaperonas Moleculares/genética , Ligação Proteica , Isomerases de Dissulfetos de Proteínas/metabolismo , Transporte ProteicoRESUMO
Human embryonic stem cells (hESCs) represent a promise for future strategies of tissue replacement. However, there are different issues that should be resolved before these cells can be used in cellular therapies; among others, the rejection of transplantable hESCs as a result of HLA incompatibility between donor cells and recipients. The hESCs exhibit a weak HLA class I expression on the cell surface, but today the responsible mechanisms are unknown. We have analyzed the level expression of HLA class I heavy chain, beta2-microglobulin (beta2-m), and antigen-processing machinery (APM) components (TAP1, TAP2, LMP2, LMP7, and Tapasin) using the HS293 hESC line by real-time quantitative RT-PCR. This analysis has revealed a low expression of beta2-m, HLA-B, and Tapasin, and an absence of expression of: TAP1, TAP2, LMP2, and LMP7 genes in the HS293 hESC line respect to the embryoid bodies (EBs) and the induced stem cells with IFNgamma (with significant differences, p<0.05). The lack or loss of HLA class I molecules due to the down-regulation of the APM components has been frequently found in tumors of different histology as specific mechanisms of immune-evasion. We described for the first time in this report that the hESCs shared similar mechanisms with respect to tumor cells responsible for the weak HLA class I expression on the cell surface.
Assuntos
Células-Tronco Embrionárias/metabolismo , Regulação da Expressão Gênica , Antígenos de Histocompatibilidade Classe I/metabolismo , Biomarcadores/metabolismo , Células-Tronco Embrionárias/citologia , Imunofluorescência , Humanos , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
Mitomycin C (MMC) treatment has been used to arrest cell proliferation but not much is known about the effect of MMC on human foreskin fibroblasts (HFF) used as feeders for human embryonic stem cells (hESC). We tested the ability of MMC to stop the proliferation of HFF and to induce apoptosis. MMC inhibited the proliferation of HFF at 10 microg/ml over 2.5h of MMC treatment showing a decrease in the proliferation index measured by Ki-67 and S and G2/M phases related to active HFF. A low percentage of cells showed necrotic or apoptotic features using different lengths of incubation. Over time, the majority of cells remained in a mitotically inactive state. The percentage of apoptotic cells increased from day 2 to day 10, at the same time as the necrotic ones increased. The HS181 hESC line grew in an undifferentiated state on inactive HFF throughout the study.
Assuntos
Apoptose/efeitos dos fármacos , Técnicas de Cocultura/métodos , Células-Tronco Embrionárias/fisiologia , Antígeno Ki-67/efeitos dos fármacos , Mitomicina/farmacologia , Ploidias , Animais , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Estudos de Viabilidade , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Citometria de Fluxo , Corantes Fluorescentes , Prepúcio do Pênis/citologia , Prepúcio do Pênis/efeitos dos fármacos , Prepúcio do Pênis/metabolismo , Humanos , Imuno-Histoquímica , Masculino , CamundongosRESUMO
La cistinuria es una aminoaciduria debida a un transporte defectuoso de cistina y de aminoácidos dibásicos (arginina, ornitina, y lisina) en la membrana apical del epitelio intestinal y túbulo proximal renal. El resultado es una ausencia de reabsorción de cistina en el túbulo proximal renal produciendo un exceso de cistina en orina y con la consiguiente formación de cálculos renales. Los cálculos de cistina son muy difíciles de eliminar por litotricia a diferencia del resto de cálculos. Por lo tanto, debería de llevarse a cabo una terapia no invasiva para prevenir la recidiva en la formación de cálculos. Esta terapia estaría basada en una alta ingesta de líquidos, alcalinización de la orina, y empleo de agentes quelantes. A la hora de preservar la función renal es necesaria la combinación de estas tres medidas terapéuticas tanto para disminuir la recurrencia como la morbilidad de la enfermedad (AU)
Cystinuria is an aminoaciduria due to the impairment of transport of cystine and dibasic amino acids (arginine, ornithine, and lysine) in the apical membrane of the intestinal epithelium and proximal renal tubule. The result is an absence of cystine reabsorption in the renal tubule producing an excess of cystine in urine and stone formation. Unlike the other stones, cystinestones are very difficult to eliminate with lithotripsy. Non invasive therapy should therefore be used to prevent relapse in stone formation. This therapy is based on the use of high fluid in take, urine alkalinization, and chelating agents. In order to preserve renal function, a combination of these three therapeutic measures is necessary to produce a low recurrence and morbidity of the disease (AU)
Assuntos
Humanos , Cistinúria/diagnóstico , Cistinúria/terapia , Cálculos Urinários/fisiopatologia , Quelantes/uso terapêutico , Ingestão de Líquidos , Citrato de Potássio/uso terapêutico , Acetazolamida/uso terapêuticoRESUMO
Human embryonic stem alls (hESC) have an unlimited capacity of proliferation and self-renewal resulting in a promise for future applications in regenerative medicine. One major problem derived from their use in cellular therapy protocols is the immunological rejection due to HLA incompatibility. Currently, there are four strategies to prevent allograft rejection of hESC; the development of a ''universal hESC line'' with lack of HLA class 1 expression; the creation of nuclear transfer hESC line; the development of hESC line banks; and the generation of haemopoietic chimerism.
Assuntos
Células-Tronco Embrionárias , Rejeição de Enxerto/prevenção & controle , Técnicas de Transferência Nuclear , Transplante de Células-Tronco , Células-Tronco Embrionárias/metabolismo , Expressão Gênica , Antígenos HLA/biossíntese , Humanos , Medicina Regenerativa/métodos , Quimeras de Transplante/metabolismo , Transplante HomólogoRESUMO
Cell line cross-contamination is a phenomenon that arises as a result of the continuous cell line culture. It has been estimated that around 20% of the cell lines are misidentified, therefore it is necessary to carry out quality control tests for the detection of this issue. Since cell line cross-contamination discovery, different methods have been applied, such as isoenzyme analysis for inter-species cross-contamination; HLA typing, and DNA fingerprinting using short tandem repeat and a variable number of tandem repeat for intra-species cross-contamination. The cell banks in this sense represent the organizations responsible for guaranteeing the authenticity of cell lines for future research and clinical uses.
RESUMO
Three analogs of the COOH-terminal fragments of human somatotropin (HGH), namely [Nle170,Ala165,182]-HGH-(150-187), [Nle170,Ala165,182]-HGH-(152-187), and [Nle170,Ala165,182]-HGH-(154-187), have been synthesized by the solid-phase method. The synthetic analogs were complemented with the natural NH2-terminal fragment [Cys(Cam)53]-HGH-(1-134) to form recombinants with HGH activities, as revealed by the rabbit liver membrane receptor binding and the Nb2 lymphoma cell assays.
Assuntos
Hormônio do Crescimento/análise , Fragmentos de Peptídeos/análise , Sequência de Aminoácidos , Fenômenos Químicos , Química , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Hormônio do Crescimento/análogos & derivados , Humanos , Focalização Isoelétrica , Prolactina/análise , Ensaio RadioliganteRESUMO
Antisera raised in rabbits against synthetic insulin-like growth factor-II (IGF-II) were used to develop a specific radioimmunoassay (RIA) for IGF-II. Affinity purified antibodies showed 6% cross-reactivity with IGF-I but failed to recognize insulin even at 10 micrograms/tube. Utilizing this RIA system, immunoreactive IGF-II was identified in the pooled samples of human follicular fluid and seminal plasma. The acid-ethanol precipitates of human seminal and follicular fluids were chromatographed on Sephadex G-50 column and the IGF-II immunoreactive fractions were subjected to reversed-phase high performance liquid chromatography. It was found that immunoactive IGF-II was eluted in the same location as that of synthetic IGF-II. The data indicate for the first time that human seminal plasma and follicular fluid contain significant amounts of IGF-II.
Assuntos
Fator de Crescimento Insulin-Like II/análise , Folículo Ovariano/metabolismo , Sêmen/análise , Somatomedinas/análise , Líquidos Corporais/análise , Cromatografia Líquida de Alta Pressão , Reações Cruzadas , Feminino , Humanos , Fator de Crescimento Insulin-Like I/análise , Masculino , RadioimunoensaioRESUMO
We describe for the first time the purification and some properties of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) isolated from anterior pituitary tissue of the African elephant (Loxodonta africana). Methodology previously applied to equine and donkey pituitaries was used to obtain purified preparations of elephant LH and FSH in yields of 8.8 and 0.48 mg, respectively, per 10 g pituitary powder. The preparations were characterized by HPLC gel filtration and amino acid analysis, both of which showed the elephant LH and FSH to be very similar to ovine LH and FSH. The preparations were also characterized by radioimmunoassays and bioassays for LH and FSH and a radioreceptor assay for FSH. Results showed virtually no cross-contamination of hormonal activities in the elephant LH and FSH preparations. Elephant LH potencies ranged from 50 to 66% of highly purified ovine LH and elephant FSH potencies ranged from 21 to 52% of highly purified ovine FSH in the various assays employed. No evidence was found for any demonstrable intrinsic FSH activity in elephant LH. The assays employed suggest possible usage for making physiological measurements of gonadotropins in the elephant.
Assuntos
Elefantes/metabolismo , Hormônio Foliculoestimulante/isolamento & purificação , Hormônio Luteinizante/isolamento & purificação , Adeno-Hipófise/análise , Aminoácidos/análise , Animais , Bioensaio , Cromatografia Líquida de Alta Pressão , Estrogênios/biossíntese , Feminino , Hormônio Foliculoestimulante/farmacologia , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/metabolismo , Células Intersticiais do Testículo/efeitos dos fármacos , Células Intersticiais do Testículo/metabolismo , Hormônio Luteinizante/farmacologia , Masculino , Radioimunoensaio , Ensaio Radioligante , Ratos , Ovinos , Testosterona/biossínteseRESUMO
Antisera raised in rabbits to a synthetic peptide consisting of 31 amino acids with a sequence identical to inhibin-like peptide (ILP) isolated from human seminal plasma afford a highly specific and sensitive radioimmunoassay. Synthetic ILP completely displaces antiserum binding of radioiodinated [Tyr4]-ILP, with half maximal displacement at 36 fmoles ILP/tube. ILP, [Tyr4]-ILP and ILP-(9-31) had essentially equal potency, while ILP-(1-25), ILP-(1-23) and ILP-(1-16) had reduced potency. No cross reactivity was found among a variety of peptide hormones and proteins. Human seminal plasma displaces 50% of [125I-Tyr4]-ILP at dilutions equivalent to 50-250 pl/tube, corresponding to immunoreactive ILP concentration of 0.5-2.5 mg/ml.
Assuntos
Inibinas , Peptídeos/análise , Sêmen/análise , Sequência de Aminoácidos , Animais , Especificidade de Anticorpos , Reações Cruzadas , Humanos , Soros Imunes , Radioisótopos do Iodo , Masculino , Coelhos/imunologia , Radioimunoensaio/métodosRESUMO
The promyelocytic leukemia (PML) protein is the product of the PML gene that fuses with the retinoic acid receptor-alpha (RARalpha) gene in acute promyelocytic leukemia (APL) and produces disruption of PML bodies. Wild-type PML localizes in the nucleus with a typical speckled pattern. PML bodies accumulate several proteins involved in multiple cellular pathways such as apoptosis, transcriptional regulation, and proteasomal degradation of ubiquitinated proteins. The ubiquitin-proteasome pathway at PML bodies is dependent on proteasome component recruitment. Proteasome components such as low-molecular weight proteins (LMPs) are frequently downregulated in different tumor tissues that present impaired major histocompatibility complex (MHC) class I expression. We have recently documented LMP7 downregulation in colorectal tumors with total loss of MHC class I antigen. An immunohistochemical study of PML protein in these tumors revealed a disrupted pattern of PML bodies in a nuclear diffuse form, as observed in APL cells. Therefore, the disruption of the PML bodies was clearly associated with LMP7 downregulation.
Assuntos
Neoplasias Colorretais/metabolismo , Antígenos de Histocompatibilidade Classe I/biossíntese , Complexos Multienzimáticos/biossíntese , Proteínas de Neoplasias/metabolismo , Proteínas Nucleares/metabolismo , Fatores de Transcrição/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Neoplasias Colorretais/patologia , Regulação para Baixo , Antígenos de Histocompatibilidade Classe I/genética , Humanos , Imuno-Histoquímica , Complexos Multienzimáticos/genética , Proteína da Leucemia Promielocítica , Complexo de Endopeptidases do Proteassoma , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Prolactin was isolated from anterior lobes of elephant pituitary glands. It consisted of 199 amino acids with three disulfide bridges and two tryptophan residues as found in prolactin from other species. The sequence of the NH2-terminal 28 amino acids was determined and shown homologous with the ovine hormone. In comparison with ovine prolactin, a marked difference was seen in the methionine content; the elephant hormone possessed only 18-34% lactogenic potency. The conformation of elephant prolactin was examined by zero order, second order and circular dichroism spectroscopy. The alpha helical content was estimated to be about 60%. In comparison with prolactins from other species, the second order spectra of elephant prolactin suggest that the local microenvironment for one or both tryptophan residues is somewhat different.