RESUMO
B cells constitute an essential line of defense from pathogenic infections through the generation of class-switched antibody-secreting cells (ASCs) in germinal centers. Although this process is known to be regulated by follicular helper T (TfH) cells, the mechanism by which B cells initially seed germinal center reactions remains elusive. We found that NKT cells, a population of innate-like T lymphocytes, are critical for the induction of B cell immunity upon viral infection. The positioning of NKT cells at the interfollicular areas of lymph nodes facilitates both their direct priming by resident macrophages and the localized delivery of innate signals to antigen-experienced B cells. Indeed, NKT cells secrete an early wave of IL-4 and constitute up to 70% of the total IL-4-producing cells during the initial stages of infection. Importantly, the requirement of this innate immunity arm appears to be evolutionarily conserved because early NKT and IL-4 gene signatures also positively correlate with the levels of neutralizing antibodies in Zika-virus-infected macaques. In conclusion, our data support a model wherein a pre-TfH wave of IL-4 secreted by interfollicular NKT cells triggers the seeding of germinal center cells and serves as an innate link between viral infection and B cell immunity.
Assuntos
Linfócitos B/imunologia , Centro Germinativo/imunologia , Imunidade Inata , Influenza Humana/imunologia , Interleucina-4/genética , Células Matadoras Naturais/imunologia , Infecção por Zika virus/imunologia , Animais , Galinhas , Cães , Centro Germinativo/citologia , Humanos , Interleucina-4/metabolismo , Macaca , Macrófagos/imunologia , Células Madin Darby de Rim Canino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
PKCß-null (Prkcb-/-) mice are severely immunodeficient. Here we show that mice whose B cells lack PKCß failed to form germinal centers and plasma cells, which undermined affinity maturation and antibody production in response to immunization. Moreover, these mice failed to develop plasma cells in response to viral infection. At the cellular level, we have shown that Prkcb-/- B cells exhibited defective antigen polarization and mTORC1 signaling. While altered antigen polarization impaired antigen presentation and likely restricted the potential of GC development, defective mTORC1 signaling impaired metabolic reprogramming, mitochondrial remodeling, and heme biosynthesis in these cells, which altogether overwhelmingly opposed plasma cell differentiation. Taken together, our study reveals mechanistic insights into the function of PKCß as a key regulator of B cell polarity and metabolic reprogramming that instructs B cell fate.
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Linfócitos B/imunologia , Diferenciação Celular/imunologia , Ativação Linfocitária/imunologia , Plasmócitos/imunologia , Proteína Quinase C beta/imunologia , Animais , Heme/biossíntese , Camundongos , Camundongos Knockout , Mitocôndrias/imunologia , Mitocôndrias/metabolismo , Plasmócitos/citologiaRESUMO
CD4(+) CD25(+) Foxp3(+) regulatory T (Treg) cells mediate immunological self-tolerance and suppress immune responses. Retinoic acid (RA), a natural metabolite of vitamin A, has been reported to enhance the differentiation of Treg cells in the presence of TGF-ß. In this study, we show that the co-culture of naive T cells from C57BL/6 mice with allogeneic antigen-presenting cells (APCs) from BALB/c mice in the presence of TGF-ß, RA, and IL-2 resulted in a striking enrichment of Foxp3(+) T cells. These RA in vitro-induced regulatory T (RA-iTreg) cells did not secrete Th1-, Th2-, or Th17-related cytokines, showed a nonbiased homing potential, and expressed several cell surface molecules related to Treg-cell suppressive potential. Accordingly, these RA-iTreg cells suppressed T-cell proliferation and inhibited cytokine production by T cells in in vitro assays. Moreover, following adoptive transfer, RA-iTreg cells maintained Foxp3 expression and their suppressive capacity. Finally, RA-iTreg cells showed alloantigen-specific immunosuppressive capacity in a skin allograft model in immunodeficient mice. Altogether, these data indicate that functional and stable allogeneic-specific Treg cells may be generated using TGF-ß, RA, and IL-2. Thus, RA-iTreg cells may have a potential use in the development of more effective cellular therapies in clinical transplantation.
Assuntos
Rejeição de Enxerto/prevenção & controle , Transplante de Pele , Pele/imunologia , Linfócitos T Reguladores/imunologia , Tretinoína/farmacologia , Transferência Adotiva , Aloenxertos , Animais , Técnicas de Cocultura , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/imunologia , Expressão Gênica , Sobrevivência de Enxerto , Interleucina-2/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ovalbumina/administração & dosagem , Pele/citologia , Baço/citologia , Baço/imunologia , Linfócitos T Reguladores/citologia , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/transplante , Fator de Crescimento Transformador beta/farmacologiaRESUMO
BACKGROUND: DNA methylation is the most important epigenetic change involved in the control of gene expression in human cells. Methylation of the p16(INK4a) gene occurs early in the development of cervical cancer. Low-grade squamous intraepithelial lesions (LSILs) are prevalent, and their behavior is variable. OBJECTIVE: To identify the HPV DNA type, detect the methylation status of the p16(INK4A) gene, and analyze their association with the cytological evolution of LSIL over a period of two years. METHODS: We conducted a cohort study with 40 participants. Cervical scrapings were collected for cytological and molecular analysis. HPV DNA detection and typing were performed by means of polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). Methylation-specific PCR was performed to detect methylation. RESULTS: HPV DNA was detected in 87% of the cases, and type 16 was the most frequent type. Methylation was detected in 11% of the cases and did not exhibit a significant correlation with the HPV type. Unfavorable cytological evolution exhibited a significant association with the presence of methylation. CONCLUSION: HPV 16 was the most frequently detected type of HPV in LSIL. Methylation of the p16(INK4A) gene was infrequent and occurred independent of the presence of HPV DNA. Methylation of the p16(INK4a) gene exhibited a significant correlation with persistence/progression of LSIL.
Assuntos
Inibidor p16 de Quinase Dependente de Ciclina/genética , Metilação de DNA/genética , Papillomavirus Humano 16/genética , Lesões Intraepiteliais Escamosas Cervicais/genética , Displasia do Colo do Útero/genética , Adulto , Biomarcadores Tumorais , Estudos de Coortes , DNA Viral/genética , Feminino , Humanos , Pessoa de Meia-Idade , Infecções por Papillomavirus/diagnóstico , Lesões Intraepiteliais Escamosas Cervicais/virologia , Adulto Jovem , Displasia do Colo do Útero/virologiaRESUMO
OBJECTIVE: To analyze the presence of HPV-DNA and TIMP-2 gene methylation in cervical precursor and invasive lesions, as well as to study the associations among the latter, the presence of HPV-DNA, and the clinical evolution of such lesions. METHODS: Cross-sectional study that includes 49 biopsy or brush smear samples from women with a normal cervix, LSIL, HSIL, microinvasive carcinoma and invasive carcinoma. The presence of HPV-DNA and specific methylation was analyzed using PCR. Thirty-eight biopsy samples for HSIL, microinvasive carcinoma and frank invasive carcinoma as well as 11 brush smear samples for LSIL and normal cervices were analyzed. RESULTS: TIMP-2 gene methylation was detected in 86.8% (33/38) of the samples from the group with lesions and 50% (4/8) of the normal samples (p=0.03). HPV-DNA was detected in 81.6% (31/38) of the samples from the group with lesions and 25% (2/8) of the normal samples (p=0.003). HPV-DNA was more frequent in the methylated samples (50%), and the group with methylation had a higher risk of unfavorable evolution than the group without methylation; however, such observations were not statistically significant (p=0.19). CONCLUSION: TIMP-2 gene methylation and the presence of HPV-DNA were characteristic of the group with cervical lesions. Methylation was not associated with the presence of HPV-DNA or an unfavorable clinical evolution.
Assuntos
Biomarcadores Tumorais/genética , Carcinoma de Células Escamosas/genética , Metilação de DNA , Infecções por Papillomavirus/genética , Inibidor Tecidual de Metaloproteinase-2/genética , Displasia do Colo do Útero/genética , Neoplasias do Colo do Útero/genética , Adulto , Idoso , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/virologia , Estudos Transversais , DNA Viral/genética , Feminino , Seguimentos , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica , Papillomaviridae/genética , Infecções por Papillomavirus/patologia , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase , Prognóstico , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia , Displasia do Colo do Útero/patologia , Displasia do Colo do Útero/virologiaRESUMO
T helper type 17 (Th17) lymphocytes are found in high frequency in tumour-burdened animals and cancer patients. These lymphocytes, characterized by the production of interleukin-17 and other pro-inflammatory cytokines, have a well-defined role in the development of inflammatory and autoimmune pathologies; however, their function in tumour immunity is less clear. We explored possible opposing anti-tumour and tumour-promoting functions of Th17 cells by evaluating tumour growth and the ability to promote tumour infiltration of myeloid-derived suppressor cells (MDSC), regulatory T cells and CD4(+) interferon-γ(+) cells in a retinoic acid-like orphan receptor γt (RORγt) -deficient mouse model. A reduced percentage of Th17 cells in the tumour microenvironment in RORγt-deficient mice led to enhanced tumour growth, that could be reverted by adoptive transfer of Th17 cells. Differences in tumour growth were not associated with changes in the accumulation or suppressive function of MDSC and regulatory T cells but were related to a decrease in the proportion of CD4(+) T cells in the tumour. Our results suggest that Th17 cells do not affect the recruitment of immunosuppressive populations but favour the recruitment of effector Th1 cells to the tumour, thereby promoting anti-tumour responses.
Assuntos
Tolerância Imunológica , Neoplasias/imunologia , Células Th1/imunologia , Células Th17/imunologia , Animais , Linhagem Celular Tumoral , Camundongos , Camundongos Mutantes , Neoplasias/genética , Neoplasias/patologia , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/genética , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/imunologia , Células Th1/patologia , Células Th17/patologiaRESUMO
One of the greatest advances in medicine during the past century is the introduction of organ transplantation. This therapeutic strategy designed to treat organ failure and organ dysfunction allows to prolong the survival of many patients that are faced with no other treatment option. Today, organ transplantation between genetically dissimilar individuals (allogeneic grafting) is a procedure widely used as a therapeutic alternative in cases of organ failure, hematological disease treatment, and some malignancies. Despite the potential of organ transplantation, the administration of immunosuppressive drugs required for allograft acceptance induces severe immunosuppression in transplanted patients, which leads to serious side effects such as infection with opportunistic pathogens and the occurrence of neoplasias, in addition to the known intrinsic toxicity of these drugs. To solve this setback in allotransplantation, researchers have focused on manipulating the immune response in order to create a state of tolerance rather than unspecific immunosuppression. Here, we describe the different treatments and some of the novel immunotherapeutic strategies undertaken to induce transplantation tolerance.
Assuntos
Rejeição de Enxerto/prevenção & controle , Fatores Imunológicos/uso terapêutico , Transplante de Órgãos , Tolerância ao Transplante/efeitos dos fármacos , Citocinas/biossíntese , Citocinas/imunologia , Células Dendríticas/imunologia , Células Dendríticas/patologia , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/patologia , Sobrevivência de Enxerto/efeitos dos fármacos , Sobrevivência de Enxerto/imunologia , Humanos , Terapia de Imunossupressão , Imunossupressores/efeitos adversos , Macrófagos/imunologia , Macrófagos/patologia , Linfócitos T/imunologia , Linfócitos T/patologia , Transplante HomólogoRESUMO
OBJECTIVE: The purpose of this study was to evaluate colposcopic sensitivity in the diagnosis of microinvasive squamous carcinoma of the cervix. STUDY DESIGN: We conducted a cross-sectional study in 151 patients from 1991-2008. The colposcopic findings of microinvasion suspicion were described by the International Federation for Cervical Pathology and Colposcopy in 2003. RESULTS: There has been colposcopic suspicion of invasion in 35 patients, which represents a sensitivity of 23%. The major colposcopic findings that were observed in the transformation zone included acetowhite epithelium in 21% (32/151 patients), coarse punctuation in 19% (29/151 patients), coarse mosaic in 17% (26/151 patients), and atypical vessels in 3.9% (6/151 patients). Suspicion of microinvasion was found in 14.5% of unsatisfactory colposcopy and in 8.6% of satisfactory colposcopy. CONCLUSION: The sensitivity of colposcopy in the diagnosis of microinvasive carcinoma of the cervix was low. Colposcopy plays an important role in directing the biopsy to the most suspicious area. The definitive diagnosis of microinvasive squamous carcinoma is established only by histologic study.
Assuntos
Carcinoma de Células Escamosas/patologia , Colposcopia , Neoplasias do Colo do Útero/patologia , Adulto , Idoso , Estudos Transversais , Feminino , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Sensibilidade e Especificidade , Adulto JovemRESUMO
Genetic transformation is crucial for both investigating gene functions and for engineering of crops to introduce new traits. Rice (Oryza sativa L.) is an important model in plant research, since it is the staple food for more than half of the world's population. As a result, numerous transformation methods have been developed for both indica and japonica rice. Since breeders continuously develop new rice varieties, transformation protocols have to be adapted for each new variety. Here we provide an optimized transformation protocol with detailed tips and tricks for a new African variety Komboka using immature embryos. In Komboka, we obtained an apparent transformation rate of up to 48% for GUS/GFP reporter gene constructs using this optimized protocol. This protocol is also applicable for use with other elite indica rice varieties.
RESUMO
OBJECTIVE: Ectonucleotide pyrophosphatase/phosphodiesterase (E-NPP) and adenosine deaminase (ADA) activities, in the platelets and serum, were examined in patients with uterine cervix neoplasia without treatment as well as in patients treated by conization or radiotherapy (RTX). DESIGN AND METHODS: The patients were divided based on the amount of time from the end of the treatments until the day of the blood sampling. Groups I (n=19) (conization) and III (n=11) (radiotherapy) (treated from one to five years earlier), groups II (n=19) (conization) and IV (n=16) (radiotherapy) (treated recently; up to three months earlier) and the non-treated group (cancer) (n=7). RESULTS: E-NPP and ADA in the platelets and E-NPP in the serum were decreased in all the treated groups in relation to the control and non-treated groups, while ADA in the serum was decreased only in the conization groups in relation to them. In group II, E-NPP and ADA, in the platelets, were increased in relation to group IV. CONCLUSION: The tendency of reduction for E-NPP and ADA indicates that they may act together to control nucleotide levels and it may also be speculated that surgery causes greater platelet activation contributing to the changes seen in the conization groups. In this sense, platelets seem to be more sensitive than serum.
Assuntos
Adenosina Desaminase/sangue , Diester Fosfórico Hidrolases/sangue , Pirofosfatases/sangue , Neoplasias do Colo do Útero/sangue , Neoplasias do Colo do Útero/enzimologia , Adulto , Animais , Plaquetas/enzimologia , Feminino , Humanos , Pessoa de Meia-Idade , Estatística como Assunto , Neoplasias do Colo do Útero/patologiaRESUMO
Ischemic stroke is a leading cause of mortality and disability particularly in the elderly. Hypertension is the most important risk factor in strokes, representing roughly 70% of all cases. Oxidative stress is believed to be one of the mechanisms taking part in neuronal damage in stroke. It is well documented that cholinergic system plays a key role in normal brain functions and in memory disturbances of several pathological processes, such as in cerebral blood flow regulation. This study investigated the oxidative status and acetylcholinesterase (AChE) activity in whole blood in patients diagnosed with acute and chronic stages of ischemia, as well as with hypertension. Malondialdehyde (MDA) levels and protein carbonylation content showed increased levels both in the acute ischemic groups and in the hypertensive group, when compared to the control. Catalase activity and reduced glutathione (GSH) levels in the acute group were also higher than in the hypertensive, chronic ischemic and control groups (p<0.05). The activity of AChE in acute ischemic patients was significantly higher than that presented by the control, hypertensive and chronic ischemic patients (p<0.05). The hypertensive group presented AChE activity significantly lower than control and chronic groups. In spite of having a defined location the ischemic event results in a systemic disorder that induces changes, which can be detected by measuring the peripheral markers of oxidative stress and AChE activity in erythrocytes.
Assuntos
Acetilcolinesterase/sangue , Isquemia Encefálica/complicações , Eritrócitos/enzimologia , Hipertensão/metabolismo , Estresse Oxidativo , Acidente Vascular Cerebral/metabolismo , Doença Aguda , Idoso , Antioxidantes/metabolismo , Catalase/sangue , Doença Crônica , Glutationa/sangue , Humanos , Peroxidação de Lipídeos , Pessoa de Meia-Idade , Carbonilação Proteica , Acidente Vascular Cerebral/etiologia , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
During B-cell activation, the dynamic reorganisation of the cytoskeleton is crucial for multiple cellular responses, such as receptor signalling, cell spreading, antigen internalisation, intracellular trafficking, and antigen presentation. However, the role of intermediate filaments (IFs), which represent a major component of the mammalian cytoskeleton, is not well defined. Here, by using multiple super-resolution microscopy techniques, including direct stochastic optical reconstruction microscopy, we show that IFs in B cells undergo drastic reorganisation immediately upon antigen stimulation and that this reorganisation requires actin and microtubules. Although the loss of vimentin in B cells did not impair B-cell development, receptor signalling, and differentiation, vimentin-deficient B cells exhibit altered positioning of antigen-containing and lysosomal associated membrane protein 1 (LAMP1+) compartments, implying that vimentin may play a role in the fine-tuning of intracellular trafficking. Indeed, vimentin-deficient B cells exhibit impaired antigen presentation and delayed antibody responses in vivo. Thus, our study presents a new perspective on the role of IFs in B-cell activation.
RESUMO
Aluminium (Al), a neurotoxic compound, has been investigated in a large number of studies both in vivo and in vitro. In this study, we investigated the effect in vivo of long-term exposure to Al on NTPDase (nucleoside triphosphate diphosphohydrolase) and 5'-nucleotidase activities in the synaptosomes (obtained from the cerebral cortex and hippocampus) and platelets of rats. Here, we investigated a possible role of platelets as peripheral markers in rats. Rats were loaded by gavage with AlCl(3) 50 mg/(kg day), 5 days per week, totalizing 60 administrations. The animals were divided into four groups: (1) control (C), (2) 50 mg/kg of citrate solution (Ci), (3) 50 mg/kg of Al plus citrate (Al+Ci) solution and (4) 50 mg/kg of Al (Al). ATP hydrolysis was increased in the synaptosomes from the cerebral cortex by 42.9% for Al+Ci and 39.39% for Al, when compared to their respective control (p<0.05). ADP hydrolysis was increased by 13.15% for both Al and Al+Ci, and AMP hydrolysis increased by 32.7% for Al and 27.25% for Al+Ci (p<0.05). In hippocampal synaptosomes, the hydrolysis of ATP, ADP and AMP, was increased by 58.5%, 28.5% and 25.92%, respectively, for Al (p<0.05) and 36.7%, 22.5% and 37.64% for Al+Ci, both when compared to their respective controls. ATP, ADP and AMP hydrolysis, in platelets, was increased by 172.3%, 188.52% and 92.1%, respectively in Al+Ci, and 317.9%, 342.8% and 177.9%, respectively, for Al, when compared to their respective controls (p<0.05). Together, these results indicate that Al increases NTPDase and 5'-nucleotidase activities, in synaptosomal fractions and platelets. Thus, we suggest that platelets could be sensitive peripheral markers of Al toxicity of the central nervous system.
Assuntos
5'-Nucleotidase/efeitos dos fármacos , Alumínio/toxicidade , Antígenos CD/efeitos dos fármacos , Apirase/efeitos dos fármacos , Plaquetas/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Sinaptossomos/efeitos dos fármacos , 5'-Nucleotidase/metabolismo , Trifosfato de Adenosina/metabolismo , Cloreto de Alumínio , Compostos de Alumínio/toxicidade , Animais , Antígenos CD/metabolismo , Apirase/metabolismo , Biomarcadores/análise , Biomarcadores/metabolismo , Plaquetas/enzimologia , Encéfalo/enzimologia , Encéfalo/fisiopatologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/enzimologia , Córtex Cerebral/fisiopatologia , Cloretos/toxicidade , Citratos/farmacologia , Hidrólise/efeitos dos fármacos , Masculino , Terminações Pré-Sinápticas/efeitos dos fármacos , Terminações Pré-Sinápticas/enzimologia , Ratos , Ratos Wistar , Citrato de Sódio , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologia , Sinaptossomos/enzimologiaRESUMO
The activities of the enzymes NTPDase (EC 3.6.1.5, apyrase, CD39) and 5'-nucleotidase (EC 3.1.3.5, CD73) were analyzed in platelets from rats submitted to demyelination by ethidium bromide (EB) and treated with interferon beta (IFN-beta). The following groups were studied: I - control (saline), II - (saline and IFN-beta), III - (EB) and IV - (EB and IFN-beta). After 7, 15 and 30 days, the animals (n=7) were sacrificed and the platelets were separated by the method of Lunkes et al. [Lunkes, G., Lunkes D., Morsch, V., Mazzanti, C., Morsch, A., Miron, V., Schetinger, M.R.C., 2004. NTPDase and 5'-nucleotidase in rats alloxan- induced diabetes. Diabetes Research and Clinical Practice 65, 1-6]. NTPDase activity for ATP and ADP substrates was significantly lower in groups II and III after seven days, when compared to control (p<0.001). At fifteen days, ATP hydrolysis was significantly lower in group III and IV and higher in group II (p<0.001), while there was an activation of ADP hydrolysis in group II (p<0.001), when compared with the control. 5'-nucleotidase activity was significantly higher in group IV (p<0.001) after seven days, and lower in the groups III and IV (p<0.001) after fifteen days in relation to the control. No significant differences were observed in NTPDase and 5'-nucleotidase activities after thirty days. In conclusion, our study demonstrated that the hydrolysis of adenine nucleotides is modified in platelets of rats demyelinated and treated with IFN-beta.
Assuntos
5'-Nucleotidase/metabolismo , Nucleotídeos de Adenina/metabolismo , Antígenos CD/metabolismo , Apirase/metabolismo , Plaquetas , Doenças Desmielinizantes , Interferon beta/uso terapêutico , Animais , Plaquetas/efeitos dos fármacos , Plaquetas/enzimologia , Plaquetas/metabolismo , Doenças Desmielinizantes/sangue , Doenças Desmielinizantes/tratamento farmacológico , Doenças Desmielinizantes/enzimologia , Modelos Animais de Doenças , Etídio , Hidrólise , Masculino , Ratos , Ratos WistarRESUMO
In this paper, we studied the influence of uremia and hemodialysis on oxidative parameters and delta-aminolevulinic acid dehydratase (delta-ALA-D) activity in control subjects, patients with chronic renal failure (CRF) on hemodialysis treatment (HD) and in patients not undergoing hemodialysis (ND). An increased lipid peroxidation was observed in the serum of HD and ND patients, as measured by the MDA serum levels. However, the level of MDA from erythrocytes was only elevated in HD patients. Blood catalase activity was increased in HD and ND groups. This study also showed a decreased activity of blood delta-aminolevulinic acid dehydratase (delta-ALA-D) in both groups of patients. This study demonstrated a positive correlation between ALA-D activity and hemoglobin, suggesting that inhibition of this enzyme might enhance anemia in CRF. A negative correlation was found between the alteration in delta-ALA-D activity and oxidative stress, which may indicate that the inhibition of ALA-D can be used as an index of oxidative stress.
Assuntos
Falência Renal Crônica/fisiopatologia , Estresse Oxidativo , Sintase do Porfobilinogênio/metabolismo , Diálise Renal , Uremia/fisiopatologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Catalase/sangue , Catalase/metabolismo , Feminino , Hemoglobinas , Humanos , Peroxidação de Lipídeos , Masculino , Pessoa de Meia-Idade , Sintase do Porfobilinogênio/sangueRESUMO
Cytokinesis is the last stage in the cell cycle. In prokaryotes, the protein FtsZ guides cell constriction by assembling into a contractile ring-shaped structure termed the Z-ring. Constriction of the Z-ring is driven by the GTPase activity of FtsZ that overcomes the energetic barrier between two protein conformations having different propensities to assemble into polymers. FtsZ is found in psychrophilic, mesophilic and thermophilic organisms thereby functioning at temperatures ranging from subzero to >100°C. To gain insight into the functional adaptations enabling assembly of FtsZ in distinct environmental conditions, we analyzed the energetics of FtsZ function from mesophilic Escherichia coli in comparison with FtsZ from thermophilic Methanocaldococcus jannaschii. Presumably, the assembly may be similarly modulated by temperature for both FtsZ orthologs. The temperature dependence of the first-order rates of nucleotide hydrolysis and of polymer disassembly, indicated an entropy-driven destabilization of the FtsZ-GTP intermediate. This destabilization was true for both mesophilic and thermophilic FtsZ, reflecting a conserved mechanism of disassembly. From the temperature dependence of the critical concentrations for polymerization, we detected a change of opposite sign in the heat capacity, that was partially explained by the specific changes in the solvent-accessible surface area between the free and polymerized states of FtsZ. At the physiological temperature, the assembly of both FtsZ orthologs was found to be driven by a small positive entropy. In contrast, the assembly occurred with a negative enthalpy for mesophilic FtsZ and with a positive enthalpy for thermophilic FtsZ. Notably, the assembly of both FtsZ orthologs is characterized by a critical concentration of similar value (1-2 µM) at the environmental temperatures of their host organisms. These findings suggest a simple but robust mechanism of adaptation of FtsZ, previously shown for eukaryotic tubulin, by adjustment of the critical concentration for polymerization.
Assuntos
Proteínas Arqueais/metabolismo , Methanocaldococcus/metabolismo , Biopolímeros/metabolismo , Escherichia coli/genética , Guanosina Trifosfato/metabolismo , Hidrólise , Cinética , Methanocaldococcus/genética , Polimerização , Temperatura , TermodinâmicaRESUMO
Vaccines remain the most effective tool to prevent infectious diseases. Here, we introduce an in vitro booster vaccination approach that relies on antigen-dependent activation of human memory B cells in culture. This stimulation induces antigen-specific B cell proliferation, differentiation of B cells into plasma cells, and robust antibody secretion after a few days of culture. We validated this strategy using cells from healthy donors to retrieve human antibodies against tetanus toxoid and influenza hemagglutinin (HA) from H1N1 and newly emergent subtypes such as H5N1 and H7N9. Anti-HA antibodies were cross-reactive against multiple subtypes, and some showed neutralizing activity. Although these antibodies may have arisen as a result of previous influenza infection, we also obtained gp120-reactive antibodies from non-HIV-infected donors, indicating that we can generate antibodies without prior antigenic exposure. Overall, our novel approach can be used to rapidly produce therapeutic antibodies and has the potential to assess the immunogenicity of candidate antigens, which could be exploited in future vaccine development.
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Autophagy is important in a variety of cellular and pathophysiological situations; however, its role in immune responses remains elusive. Here, we show that among B cells, germinal center (GC) cells exhibited the highest rate of autophagy during viral infection. In contrast to mechanistic target of rapamycin complex 1-dependent canonical autophagy, GC B cell autophagy occurred predominantly through a noncanonical pathway. B cell stimulation was sufficient to down-regulate canonical autophagy transiently while triggering noncanonical autophagy. Genetic ablation of WD repeat domain, phosphoinositide-interacting protein 2 in B cells alone enhanced this noncanonical autophagy, resulting in changes of mitochondrial homeostasis and alterations in GC and antibody-secreting cells. Thus, B cell activation prompts a temporal switch from canonical to noncanonical autophagy that is important in controlling B cell differentiation and fate.
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Autofagia/imunologia , Linfócitos B/imunologia , Linfócitos B/virologia , Viroses/imunologia , Animais , Regulação para Baixo , Centro Germinativo/imunologia , Centro Germinativo/virologia , Ativação Linfocitária , Alvo Mecanístico do Complexo 1 de Rapamicina , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Mitocôndrias/metabolismo , Complexos Multiproteicos/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Repetições WD40/genéticaRESUMO
BACKGROUND: Cervical cancer is a major cause of morbidity among women. We investigated the treatment effect on oxidative status from patients submitted to radiotherapy or conization surgery to high-grade SIL (squamous intraepithelial lesion) treatment, and oxidative profile from patients newly diagnosed for uterine cervix cancer, without treatment. METHODS: We determined the catalase activity in blood, reduced glutathione (GSH) in plasma, TBARS and protein carbonyl content from serum samples of the patients. RESULTS: The catalase activity, GSH levels, TBARS and protein carbonyl content had no statistical differences related to the controls, neither when the 2 treatments were compared, possibly because the antioxidant defense may be acting in the first period of the neoplasic transformation, and maybe indicating a possible arrest of the tumor cells caused by the efficiency of the treatments. In the non-treated patients, TBARS and protein carbonyl contents, GSH levels and catalase activity were shown to be increased comparing with the treated patients and compared with the controls indicating an tumor effect on oxidative profile, and the antioxidant activity been increased in the beginning of the tumor development. CONCLUSIONS: We suggest that the treatments were efficient in arrest of the tumor.
Assuntos
Biomarcadores/sangue , Carcinoma de Células Escamosas/sangue , Displasia do Colo do Útero/sangue , Neoplasias do Colo do Útero/sangue , Adulto , Idoso , Proteínas Sanguíneas/metabolismo , Carcinoma de Células Escamosas/radioterapia , Carcinoma de Células Escamosas/cirurgia , Catalase/sangue , Conização , Feminino , Glutationa/sangue , Humanos , Peroxidação de Lipídeos/efeitos da radiação , Pessoa de Meia-Idade , Oxirredução/efeitos da radiação , Carbonilação Proteica/efeitos da radiação , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Neoplasias do Colo do Útero/radioterapia , Neoplasias do Colo do Útero/cirurgia , Displasia do Colo do Útero/radioterapia , Displasia do Colo do Útero/cirurgiaRESUMO
O estudo teve por objetivo conhecer a visão e a prática de professores de ciências e alunos de curso de licenciatura em Ciências Biológicas em relação aos distúrbios de aprendizagem e ao fracasso escolar, de modo a compreender os sentidos e significados construídos por eles em relação à crescente atribuição de responsabilidade biológica ao suposto fracasso no ensino. Trata-se de pesquisa qualitativa, que se utilizou de aplicação de questionário e entrevista semiestruturada com professores de escolas pública e particular e com alunos de um curso de graduação/licenciatura de uma universidade pública. Como resultados, é possível identificar a sobreposição em relação ao entendimento e uso de terminologias como "distúrbios", "problemas" e "dificuldades" de aprendizagem, sendo utilizadas pelos professores e estudantes participantes da pesquisa, como sinônimos para designar um processo análogo. Verificou-se a atribuição de causa biológica a qualquer dificuldade ou problema de aprendizado do aluno, ainda que a causa seja, de fato, devido a fatores sociais ou psicológicos. Evidenciou-se o despreparo para com a realização do diagnóstico, bem como desconhecimento em relação às formas de se fazer, atribuindo esse papel a outros profissionais que acreditam estar mais preparados para lidar com esses casos, considerando-se que o tratamento medicamentoso possa ser o mais efetivo. Desse modo, os resultados obtidos demonstram a importância de investigações e elucidações mais profundas a respeito do cotidiano escolar no que se refere às questões atreladas ao processo ensino-aprendizagem e à crescente medicalização de crianças e adolescentes.
The purpose of this study was to understand the vision and practice of science teachers and students of a licentiate degree course in Biological Sciences in relation to learning disorders and school failure, in order to understand the senses and meanings they constructed in relation to the increasing attribution of biological responsibility to the supposed school failure. It is a qualitative research, in which was used a questionnaire application and a semi-structured interview with public and private school teachers and with students of a undergraduate/licentiate course from a public university. As results it is possible to identify the overlap in terms of understanding and use of terminologies such as "disturbs", "problems" and "difficulties" of learning, by teachers and students participating in the research, as synonyms to designate the same process. The attribution of biological cause to any difficulty or learning problem of the student has been verified, even if the cause is, in fact, due to social or psychological factors. The lack of preparation for diagnosis was evidenciated, as well as lack of knowledge about the ways of doing it, attributing this role to other professionals who believe that they are better prepared to assist these cases, considering that drug treatment may be the most effective. Thus, the results obtained demonstrate the importance of investigations and more profound elucidations about the school daily life in relation to the issues linked with the teaching-learning process and the increasing medicalization of children and adolescents.
El estudio tuvo como objetivo conocer la visión y la práctica de los profesores de ciencias y los estudiantes de la licenciatura en Ciencias Biológicas en relación a los trastornos de aprendizaje y fracaso escolar, con el fin de comprender los significados construidos por ellos en relación al crecente aumento de la asignación de la responsabilidad biológica al supuesto fracaso en la enseñanza. Se trata de una investigación cualitativa, en la cual se utilizo la aplicación de cuestionarios y entrevistas semiestructuradas a maestros de escuelas públicas y privadas y a estudiantes en un curso de grado / licenciatura de una universidad pública. Como resultado, es posible identificar la superposición en relación con la comprensión y el uso de terminología como "trastornos", "problemas" y "dificultades" de aprendizaje, siendo utilizados por los profesores y estudiantes que participaron de la encuesta, indistintamente para describir un mismo proceso. Se encontró la asignación de causa biológica a cualquier dificultad o problema de aprendizaje de los estudiantes, aun que la causa sea de hecho, debido a factores sociales o psicológicos. La falta de preparación para realización de diagnóstico se hizo evidente, así como el desconocimiento de formas de hacer, asignando ese papel a otros profesionales que creen estar mejor preparados para hacer frente a estos casos, teniendo en cuenta que el tratamiento farmacológico puede ser más eficaz. Por lo tanto, los resultados demuestran la importancia investigaciones y mejores esclarecimiento sobre el cotidiano de la escuela cuando se trata de cuestiones relacionadas al proceso de enseñanza-aprendizaje y la creciente medicalización de niños y adolescentes.