RESUMO
AIMS: Bipolar disorder is associated with premature mortality, but evidence is mostly derived from Western countries. There has been no research evaluating shortened lifespan in bipolar disorder using life-years lost (LYLs), which is a recently developed mortality metric taking into account illness onset for life expectancy estimation. The current study aimed to examine the extent of premature mortality in bipolar disorder patients relative to the general population in Hong Kong (HK) in terms of standardised mortality ratio (SMR) and excess LYLs, and changes of mortality rate over time. METHODS: This population-based cohort study investigated excess mortality in 12 556 bipolar disorder patients between 2008 and 2018, by estimating all-cause and cause-specific SMRs, and LYLs. Trends in annual SMRs over the 11-year study period were assessed. Study data were retrieved from a territory-wide medical-record database of HK public healthcare services. RESULTS: Patients had higher all-cause [SMR: 2.60 (95% CI: 2.45-2.76)], natural-cause [SMR: 1.90 (95% CI: 1.76-2.05)] and unnatural-cause [SMR: 8.63 (95% CI: 7.34-10.03)] mortality rates than the general population. Respiratory diseases, cardiovascular diseases and cancers accounted for the majority of deaths. Men and women with bipolar disorder had 6.78 (95% CI: 6.00-7.84) years and 7.35 (95% CI: 6.75-8.06) years of excess LYLs, respectively. The overall mortality gap remained similar over time, albeit slightly improved in men with bipolar disorder. CONCLUSIONS: Bipolar disorder is associated with increased premature mortality and substantially reduced lifespan in a predominantly Chinese population, with excess deaths mainly attributed to natural causes. Persistent mortality gap underscores an urgent need for targeted interventions to improve physical health of patients with bipolar disorder.
Assuntos
Transtorno Bipolar , Transtorno Bipolar/epidemiologia , Causas de Morte , Estudos de Coortes , Feminino , Hong Kong/epidemiologia , Humanos , Expectativa de Vida , MasculinoRESUMO
The success of treatment for malignancies, especially those undergoing radiation therapy or chemotherapy, has long been recognized to depend on the degree of hypoxia in the tumor. In addition to the prognostic value of knowing the tumor's initial level of hypoxia, assessing the tumor oxygenation during standard therapy or oxygen-related treatments (such as breathing oxygen-enriched gas mixtures or taking drugs that can increase oxygen supply to tissues) can provide valuable data to improve the efficacy of treatments. A series of early clinical studies of tumors in humans are ongoing at Dartmouth and Emory using electron paramagnetic resonance (EPR) oximetry to assess tumor oxygenation, initially and over time during either natural disease progression or treatment. This approach has the potential for reaching the long-sought goal of enhancing the effectiveness of cancer therapy. In order to effectively reach this goal, we consider the validity of the practical and statistical assumptions when interpreting the measurements made in vivo for patients undergoing treatment for cancer.
Assuntos
Neoplasias , Oximetria , Oxigênio , Hipóxia Tumoral , Espectroscopia de Ressonância de Spin Eletrônica , Humanos , Neoplasias/metabolismo , Oxigênio/metabolismoRESUMO
BACKGROUND: Approximately 10% of severely obese bariatric-surgery-seeking individuals report a lifetime history of suicide attempts, a higher rate than in the general community. Being overweight is associated with weight-related stigma, making an individual more vulnerable to social isolation, a potential risk factor for suicidal ideation and/or behavior. AIMS: In this cross-sectional study of surgery-seeking adults with severe obesity, we examined whether weight-related stigma increases (1) the likelihood of suicidal ideation and/or behavior or (2) the degree of loneliness; and whether hypotheses (1) and (2) are supported (3) if loneliness mediates the effect of weight-related stigma on suicidal ideation and/or behavior. METHODS: Online questionnaires were administered to 301 women and 95 men seeking bariatric surgery. RESULTS: Approximately 30.3% reported having at least a passing thought of suicide, and 5.55% a suicide attempt during their lifetime. The suicide attempt rate appears lower than other bariatric surgery samples, but possibly higher than community and other surgery sample rates. For severely obese surgery-seeking women, weight-related stigma was associated with suicidal ideation and/or behavior, though this was not mediated by loneliness. CONCLUSIONS: Future studies are needed to model and compare suicidal ideation and/or behavior in bariatric-surgery-seeking individuals and control groups.
Assuntos
Cirurgia Bariátrica/psicologia , Solidão/psicologia , Obesidade Mórbida/psicologia , Ideação Suicida , Tentativa de Suicídio/estatística & dados numéricos , Adulto , Estudos Transversais , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Fatores Sexuais , Inquéritos e QuestionáriosRESUMO
Inflammatory processes are thought to underlie the dopamine (DA) neuron loss seen in Parkinson's disease (PD). However, it is not known if the inflammation precedes that loss, or is a consequence of it. We injected tumor necrosis factor alpha (TNFalpha) and interleukin 1 beta (IL-1beta) into the median forebrain bundle to determine if these pro-inflammatory cytokines could induce DA neuron loss in the substantia nigra (SN) by themselves. The magnitude of the DA cell loss as well as the decreases in striatal DA, were both dose and time to sacrifice dependent. Injecting both cytokines together produced greater cell losses and DA reductions than that seen when the cytokines were injected alone. The DA neuron loss seen was more pronounced in the lateral nigra and its ventral tier and similar to that seen when other toxins are injected. These data suggest that TNFalpha and IL-1beta can induce DA neuron loss by themselves and could produce DA neuron loss independent of other inflammatory events.
Assuntos
Dopamina/metabolismo , Interleucina-1/administração & dosagem , Neurônios/efeitos dos fármacos , Substância Negra/efeitos dos fármacos , Fator de Necrose Tumoral alfa/administração & dosagem , Animais , Morte Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Injeções Intraventriculares , Masculino , Neurônios/patologia , Ratos , Substância Negra/patologiaRESUMO
The present study was conducted to identify the localization and possible contribution of the two estrogen receptor (ER) subtypes in the rat brain at postnatal (P) days 3, 7 and 14. Evaluation of the distribution of ERalpha and ERbeta immunoreactive (ir) nuclei did not reveal gender differences at the developmental point times examined. With the exception of the cerebral cortex, the pattern of staining for these ERs was unchanged across the postnatal ages examined. The distribution of ERalpha-ir nuclei was wider than ERbeta-ir during brain development. From P3, ERbeta and ERalpha-ir nuclei were found in different regions of the cerebral cortex, basal forebrain, amygdala, thalamus, hypothalamus, mesencephalon, pons, cerebellum and medulla oblongata. In addition, ERalpha-ir nuclei were exclusively detected in the hippocampal subfields, epithalamus and in several circumventricular organs. ERalpha and ERbeta dual immunofluorescence revealed positive nuclei in the medial part of the bed nucleus of the stria terminalis, periventricular preoptic nucleus and in caudal aspects of the ventrolateral part of the ventromedial hypothalamic nucleus. Although the functional significance of the dual expression of both ERs within the same nuclei remains unknown, it is possible that ERs play different roles in gene regulation within the same cell. The presence of ERs in diverse brain regions through early postnatal periods supports a potential role for estrogens in neural differentiation.
Assuntos
Encéfalo/metabolismo , Neurônios/metabolismo , Receptores de Estrogênio/metabolismo , Envelhecimento , Animais , Animais Recém-Nascidos , Encéfalo/anatomia & histologia , Encéfalo/crescimento & desenvolvimento , Receptor alfa de Estrogênio , Receptor beta de Estrogênio , Feminino , Imuno-Histoquímica , Masculino , Neurônios/citologia , Ratos , Ratos Endogâmicos F344Assuntos
Bile/diagnóstico por imagem , Doenças da Vesícula Biliar/diagnóstico por imagem , Vesícula Biliar/diagnóstico por imagem , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Cólica/etiologia , Doenças da Vesícula Biliar/etiologia , Humanos , Icterícia/etiologia , UltrassonografiaRESUMO
Hypoxia initiates numerous intracellular signaling pathways important in regulating cell proliferation, differentiation, and death. In this study, we investigated the pathway that hypoxia uses to activate Akt and inactivate glycogen synthase kinase-3 (GSK-3), two proteins the functions of which are important in cell survival and energy metabolism. Severe hypoxia (0.01% oxygen) initiated a signaling cascade by inducing the tyrosine phosphorylation of the platelet-derived growth factor (PDGF) receptor within 1 h of treatment and increasing receptor association with the p85 subunit of phosphatidylinositol 3-kinase (PI 3-K). Hypoxia-induced signaling also resulted in PI 3-K-dependent phosphorylation of Akt on Ser-473, a modification of Akt that is important for its activation. This activation of Akt by hypoxia was substantially diminished in cells that possessed mutations in their PDGF receptor-PI 3-K interaction domain. In addition, Akt activation by hypoxia was resistant to treatment with the growth factor receptor poison suramin but was sensitive to treatment with the PI 3-K inhibitor wortmannin. Activation of Akt by hypoxia resulted in the phosphorylation of GSK-3alpha and GSK-3beta at Ser-9 and Ser-21, two well-documented Akt phosphorylation sites, respectively, that are inactivating modifications of each GSK-3 isoform. In support of the phosphorylation data, GSK-3 activity was significantly reduced under hypoxia. In conclusion, we propose that hypoxia activates a growth factor receptor/PI 3-K/Akt cascade that leads to GSK-3 inactivation, a pathway that can impact cell survival, proliferation, and metabolism.
Assuntos
Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Oxigênio/fisiologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Serina-Treonina Quinases , Proteínas Proto-Oncogênicas/metabolismo , Receptor beta de Fator de Crescimento Derivado de Plaquetas/metabolismo , Transdução de Sinais/fisiologia , Androstadienos/farmacologia , Animais , Proteínas Quinases Dependentes de Cálcio-Calmodulina/antagonistas & inibidores , Hipóxia Celular/fisiologia , Linhagem Celular , Cães , Ativação Enzimática , Inibidores Enzimáticos/farmacologia , Células Epiteliais/enzimologia , Fibrossarcoma/enzimologia , Quinase 3 da Glicogênio Sintase , Quinases da Glicogênio Sintase , Humanos , Rim/citologia , Rim/enzimologia , Fosforilação , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt , Transdução de Sinais/efeitos dos fármacos , Células Tumorais Cultivadas , Tirosina/metabolismo , WortmaninaRESUMO
The comparison of the genomes of two very closely related human mucosal pathogens, Mycoplasma genitalium and Mycoplasma pneumoniae, has helped define the essential functions of a self-replicating minimal cell, as well as what constitutes a mycoplasma. Here we report the complete sequence of a more distant phylogenetic relative of those bacteria, Ureaplasma urealyticum (parvum biovar), which is also a mucosal pathogen of humans. It is the third mycoplasma to be sequenced, and has the smallest sequenced prokaryotic genome except for M. genitalium. Although the U. urealyticum genome is similar to the two sequenced mycoplasma genomes, features make this organism unique among mycoplasmas and all bacteria. Almost all ATP synthesis is the result of urea hydrolysis, which generates an energy-producing electrochemical gradient. Some highly conserved eubacterial enzymes appear not to be encoded by U. urealyticum, including the cell-division protein FtsZ, chaperonins GroES and GroEL, and ribonucleoside-diphosphate reductase. U. urealyticum has six closely related iron transporters, which apparently arose through gene duplication, suggesting that it has a kind of respiration system not present in other small genome bacteria The genome is only 25.5% G+C in nucleotide content, and the G+C content of individual genes may predict how essential those genes are to ureaplasma survival.
Assuntos
Genoma Bacteriano , Ureaplasma urealyticum/genética , Trifosfato de Adenosina/metabolismo , Evolução Biológica , DNA Bacteriano , Humanos , Ferro/metabolismo , Dados de Sequência Molecular , Análise de Sequência de DNA , Ureaplasma urealyticum/classificação , Ureaplasma urealyticum/metabolismo , Ureaplasma urealyticum/patogenicidadeRESUMO
Tumor angiogenesis, the development of new blood vessels during malignant progression, is a regulated process that has both genetic and physiological controls. Physiologically, angiogenesis is stimulated by decreases in tissue oxygenation (i.e., hypoxia). We investigated the effect of hypoxia on the expression of two angiogenic factors reported to be genetically regulated by the p53 tumor suppressor gene: (a) the angiogenic inhibitor thrombospondin 1 (TSP-1); and (b) the angiogenic inducer vascular endothelial growth factor (VEGF). Analysis of rodent cells that differ in their p53 genotype (p53+/+ or p53-/-) indicated that in vitro exposure to hypoxia simultaneously suppressed TSP-1 and induced VEGF expression, regardless of the p53 genotype. On transformation of these cells with E1A and oncogenic H-ras, the basal level of TSP-1 expression was strongly diminished, whereas that of VEGF could still be induced by hypoxia. Consistent with these in vitro findings, sections of tumors derived from the transformed p53+/+ and p53-/- cells showed that VEGF protein overlapped with regions of hypoxia, whereas TSP-1 protein was below the limits of detection in tumor tissue. Using a panel of normal/immortalized and transformed human cells, it was found that the ability of hypoxia to inhibit TSP-1 expression depends on the cell type and/or the degree of transformation. In contrast, VEGF expression was induced by hypoxia in all of the human cell types examined. Together, these findings suggest that hypoxic and oncogenic signals could interact in the tumor microenvironment to inhibit TSP-1 and induce VEGF expression, promoting the switch to the angiogenic phenotype.
Assuntos
Carcinoma de Células Escamosas/genética , Hipóxia Celular , Fatores de Crescimento Endotelial/genética , Regulação da Expressão Gênica , Genes p53 , Linfocinas/genética , Trombospondina 1/genética , Neoplasias do Colo do Útero/genética , Animais , Carcinoma de Células Escamosas/patologia , Divisão Celular , Células Cultivadas , Feminino , Humanos , Camundongos , Camundongos SCID , Transfecção , Proteína Supressora de Tumor p53/genética , Neoplasias do Colo do Útero/patologia , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio Vascular , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Inefficient maturation and trafficking to the cell surface of the cystic fibrosis transmembrane conductance regulator (CFTR) is the primary cause of cystic fibrosis. CFTR protein that fails to mature accumulates as an immature core-glycosylated protein and is rapidly degraded. To determine how the structures of mature and immature CFTR are different, we compared the properties of CFTR that had been expressed in the presence or absence of the proteasome inhibitor, MG-132 (carbobenzoxy-L-leucyl-L-leucyl-L-leucinal). Transient expression of wild-type CFTR in the presence of submicromolar concentrations of MG-132 blocks maturation of the protein. We found that expression of CFTR in the presence of MG-132 trapped the protein in a trypsin-sensitive conformation. In addition, the structure of the carboxyl-terminus of immature and mature CFTR differed as histidine-tagged mature CFTR was preferentially recovered by metal-chelate chromatography. No chloride channel activity was detected when membranes containing immature CFTR were fused with planar lipid bilayers. These results show that expression of CFTR in the presence of MG-132 traps the protein in an altered conformation that may be inactive.
Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/antagonistas & inibidores , Regulador de Condutância Transmembrana em Fibrose Cística/química , Processamento de Proteína Pós-Traducional , Sequência de Aminoácidos , Animais , Células COS , Linhagem Celular , Inibidores de Cisteína Proteinase/farmacologia , Regulador de Condutância Transmembrana em Fibrose Cística/biossíntese , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Relação Dose-Resposta a Droga , Retículo Endoplasmático/efeitos dos fármacos , Retículo Endoplasmático/enzimologia , Retículo Endoplasmático/genética , Retículo Endoplasmático/metabolismo , Humanos , Leupeptinas/farmacologia , Manosil-Glicoproteína Endo-beta-N-Acetilglucosaminidase/metabolismo , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Mutação Puntual , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Processamento de Proteína Pós-Traducional/genética , Tripsina/metabolismoRESUMO
OBJECTIVE: To explore the relationship between the properties of compliance and the change of structure of components in anastomosed arteries. METHODS: The arterial pressure and diameter of femoral arteries of dogs were measured in vivo before and after arterial anastomosis in different time intervals to deduce the arterial compliance. The anastomosed arteries were removed and evaluated through light microscopic examination and various staining methods, the relative contents of elastin, collagen and smooth muscles were measured through image analysis system. RESULTS: The compliance of arteries was gradually decreased after anastomosis with peak-time on the 14th day. The content of elastin at different time had no significant difference, while the content of collagen increased gradually, the ratio of them was increased. CONCLUSION: The property of compliance of anastomosed arteries is closely related to the contents of the structural components.
Assuntos
Artéria Femoral/patologia , Artéria Femoral/cirurgia , Anastomose Cirúrgica , Animais , Colágeno/metabolismo , Complacência (Medida de Distensibilidade) , Cães , Feminino , Artéria Femoral/metabolismo , Masculino , Músculo Liso Vascular/patologiaRESUMO
Intrathymic injection of the Abelson murine leukemia virus (A-MuLV) results in transformation of immature T and B lymphoid cells. In this report we demonstrate that the concentration of A-MuLV injected into murine thymi influences the selection of the transformation target. Thus, concentrated A-MuLV gives rise to Thy-1+ B220- thymomas. In contrast, dilute virus induces B220+ thymomas that also express low levels of Thy-1 (Thy-1lo), a phenotype that is similar to marrow-derived progenitor B-lymphoid cells (pro-B cells) that are highly susceptible to A-MuLV transformation in vitro. However, rare B220+ lymphoid cells isolated from normal adult thymi were not transformed by A-MuLV in vitro, while B220+ cells isolated from bone marrow were highly susceptible to transformation by A-MuLV. The Thy-1lo B220+ population in the primary thymomas had not rearranged TCRgamma, TCRbeta, or Igkappa genes, but contained subpopulations that assembled Ig DJ(H) or VDJ(H) genes and were therefore similar to transformed pro- and pre-B cells obtained from A-MuLV infected fetal liver and adult bone marrow, respectively. However, unlike A-MuLV-transformed pro- and pre-B cells, many (40-70%) of the Thy-1lo B220+ transformed thymoma cells had not rearranged Igh genes, and therefore appear to represent undifferentiated lymphoid cells. We conclude that A-MuLV may transform an undifferentiated lymphoid target in the thymus.
Assuntos
Vírus da Leucemia Murina de Abelson , Transformação Celular Viral , Antígenos Comuns de Leucócito/isolamento & purificação , Antígenos Thy-1/isolamento & purificação , Timoma/etiologia , Timo/citologia , Animais , Feminino , Rearranjo Gênico do Linfócito T , Genes de Imunoglobulinas , Células-Tronco Hematopoéticas , Linfócitos/virologia , Camundongos , Camundongos Endogâmicos BALB C , Fenótipo , Timoma/imunologia , Timoma/patologia , Timo/virologiaRESUMO
Recent studies have suggested the importance of hypoxia-inducible transcription factors in development, yet the questions of whether hypoxia actually exists in a developing embryo in vivo and, if so, what role it plays in development remain unanswered. In this study, we directly demonstrate that regions of hypoxia, most prominently the hindbrain, otic vesicle, and first branchial arch, exist in a gestational day (GD) 11 rat embryo grown in utero. We also show that varying the oxygen environment of an embryo affects its morphological development. Rat embryos which were grown at 45% oxygen from GD 9-11 showed gross morphological abnormalities, including defective cranial neural tube closure, incomplete otic vesicle invagination, and abnormal somite formation and embryo turning. These embryos, in addition, exhibited reduced cell death. On the other hand, embryos which were grown at 5% oxygen during the same period were stunted in overall growth, yet morphologically normal, and displayed prominent areas of apoptosis. In this study, we propose that embryonic development, like tumor development, requires two different but interactive sets of signals. One set exists in the genetic program for development; the other set arises from changes in the microenvironment of the embryo. Therefore, it is the interplay between these two sets of cues that drives normal embryonic development. The requirement for hypoxia to activate apoptotic cell death is but one example of such interactions.
Assuntos
Apoptose/fisiologia , Encéfalo/embriologia , Embrião de Mamíferos/fisiologia , Desenvolvimento Embrionário e Fetal/fisiologia , Hipóxia , Animais , Apoptose/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Embrião de Mamíferos/citologia , Desenvolvimento Embrionário e Fetal/efeitos dos fármacos , Feminino , Microcefalia/etiologia , Morfogênese/efeitos dos fármacos , Defeitos do Tubo Neural/etiologia , Oxigênio/farmacologia , Gravidez , Ratos , Ratos Sprague-DawleyRESUMO
The deposition of beta-amyloid within the entorhinal cortex (EC) may play a key role in the development of mild cognitive impairment (MCI) in the elderly. To examine the relationship of beta-amyloid deposition to MCI, EC tissue immunostained for this protein was quantitated from a cohort of aged Catholic religious clergy with a clinical diagnosis of MCI and compared to those with no cognitive impairment (NCI) and Alzheimer's disease (AD). beta-amyloid staining was seen in 12 of the 20 NCI, in 10 of 12 MCI, and in all 12 AD cases within the EC. beta-amyloid immunoreactivity displayed two patterns within the EC: (1) a crescent-shaped band within layers 3-4 or (2) bilaminar staining mainly within layers 2-3 and 5-6. Ten cases failed to display any detectable beta-amyloid imunoreactivity. Despite the heterogeneity of beta-amyloid loads within the clinical groups, decomposing an analysis of variance revealed a significant difference across groups in mean beta-amyloid load within the EC based upon a linear trend analysis. Multiple comparisons testing revealed that NCI individuals had a significantly lower mean beta-amyloid load (1.32) than AD individuals (4.55). The MCI individuals had a mean intermediate (2.60) load between NCI and AD, but not statistically distinguishable from the mean for either NCI or AD. Spearman rank correlation showed a trend for decreasing MMSE with increasing amyloid load that failed to reach statistical significance. Since many NCI cases displayed beta-amyloid loads equal to or greater than that seen in some MCI and some AD cases, it is mostly likely that deposition of this protein is not the sole pathogenic event underlying cognitive impairment in the elderly.
Assuntos
Doença de Alzheimer/patologia , Peptídeos beta-Amiloides/análise , Transtornos Cognitivos/patologia , Córtex Entorrinal/patologia , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/genética , Apolipoproteínas E/genética , Autopsia , Catolicismo , Transtornos Cognitivos/genética , Córtex Entorrinal/química , Feminino , Genótipo , Humanos , Masculino , Estados UnidosRESUMO
Many forms of acute brain injury are associated with a secondary, glutamate- and calcium-dependent cascade which greatly exacerbates the final damage. The calcium-dependent protease, calpain, has been implicated as an important variable in this pathogenic process. The present studies tested (i) if similar secondary degeneration occurs following surgical ablation of a discrete area within rat visual cortex, (ii) if activation of calpain contributes to the secondary degeneration by spreading into areas adjacent to the ablation, and (iii) if blocking calpain's proteolytic effects reduces the secondary degeneration attendant to the lesion. Antibodies selective for a protein fragment specifically generated by calpain were used to map areas in which the protease had been activated. Labeling was present 5 min after surgery in a narrow zone surrounding the ablated region. The volume of the immunopositive staining increased twofold within 24 h and fivefold by 48 h, at which time it was equivalent in size to the original lesion. This staining pattern significantly decreased in size at 5 days postsurgery. Application of calpain inhibitors to the ablation site immediately after surgery reduced the spread of calpain activation by approximately 80%. Following cortical ablation, the volume of the lateral geniculate nucleus ipsilateral to the cortical ablation shrank by 46 +/- 3% in control rats but only by 31 +/- 5% in animals given the calpain inhibitors. These results establish that (i) a secondary degenerative cascade is unleashed following discrete cortical surgery which expands into brain areas clearly outside the initial perturbation site, (ii) the gradual expansion of calpain activation contributes to the underlying secondary pathology, and (iii) blocking calpain activity substantially reduces atrophy in areas anatomically connected, but physically distal to the damaged zone. The possible utility of topical applications of calpain inhibitors, or analogously acting drugs, in minimizing the secondary effects of brain surgery is discussed.
Assuntos
Cálcio/fisiologia , Calpaína/antagonistas & inibidores , Córtex Cerebral/patologia , Inibidores de Cisteína Proteinase/farmacologia , Dipeptídeos/farmacologia , Animais , Anticorpos , Calpaína/imunologia , Córtex Cerebral/lesões , Corpos Geniculados/patologia , Imuno-Histoquímica , Masculino , Degeneração Neural/patologia , Ratos , Ratos Sprague-Dawley , Espectrina/imunologia , Espectrina/metabolismo , Córtex Visual/patologiaRESUMO
We have previously demonstrated that fetal nigral grafts can survive, reinnervate the striatum, and mediate clinically relevant recovery in a patient with Parkinson's disease (PD). Most previous autopsy cases have failed to identify meaningful numbers of viable grafted cells suggesting that differences in critical transplant variables determine graft viability. The present study evaluated the structural and functional correlates of fetal nigral transplantation in a second PD patient who received fetal nigral grafts according to our previously published transplant protocol. A 61-year-old woman with severe PD received bilateral fetal nigral grafts to the postcommissural putamen from seven donor fetuses (four right side and three left side) aged 6.5-9 weeks postconception. This patient died 19 months after surgery from a cause unrelated to the transplant surgery. Her postoperative clinical course was characterized by improved motor and activities of daily living scores during "off time," reduced "off time," and increased "on" time without dyskinesia. Positron emission tomography (PET) scans revealed a bilateral and progressive increase in fluorodopa (FD) uptake within the grafted putamen. Postmortem examination of the right hemisphere revealed large oval-shaped grafts containing more than 138,000 tyrosine-hydroxylase-immunoreactive (TH-ir) neurons. Grafted cells formed a seamless border with the host and provided dense TH-ir innervation to 78% of the host postcommissural putamen. Graft-mediated sprouting of host fibers was not observed. These data provide essential confirmation that, under appropriate transplant conditions, grafted nigral neurons can survive, reinnervate the host striatum, and provide clinical benefit to PD patients. These findings also support the concept that improved motor function and striatal FD uptake on PET after nigral grafting in PD are the result of the viability of grafted neurons and graft-derived reinnervation of the host striatum.
Assuntos
Transplante de Tecido Encefálico/fisiologia , Transplante de Tecido Fetal/fisiologia , Doença de Parkinson/cirurgia , Substância Negra/transplante , Sobrevivência de Tecidos/fisiologia , Transplante de Tecido Encefálico/patologia , Corpo Estriado/patologia , Corpo Estriado/fisiopatologia , Feminino , Transplante de Tecido Fetal/patologia , Seguimentos , Idade Gestacional , Humanos , Processamento de Imagem Assistida por Computador , Técnicas Imunoenzimáticas , Pessoa de Meia-Idade , Regeneração Nervosa/fisiologia , Exame Neurológico , Doença de Parkinson/patologia , Doença de Parkinson/fisiopatologia , Putamen/patologia , Putamen/fisiopatologia , Substância Negra/embriologia , Tomografia Computadorizada de Emissão , Tirosina 3-Mono-Oxigenase/fisiologiaRESUMO
EGF-responsive neural stem cells isolated from murine striatum have the capacity to differentiate into both neurons and glia in vitro. Genetic modification of these cells is hindered by a number of problems such as gene stability and transfection efficiency. To circumvent these problems we generated transgenic mice in which the human GFAP promoter directs the expression of human NGF. Neural stem cells isolated from the forebrain of these transgenic animals proliferate and form clusters, which appear identical to stem cells generated from control animals. Upon differentiation in vitro, the transgenic stem cell-derived astrocytes express and secrete bioactive hNGF. Undifferentiated GFAP-hNGF or control stem cells were transplanted into the striatum of adult rats. One and 3 weeks after transplantation, hNGF was detected immunocytochemically in an halo around the transplant sites. In GFAP-hNGF-grafted animals, intrinsic striatal neurons proximal to the graft appear to have taken up hNGF secreted by the grafted cells. Ipsilateral to implants of GFAP-hNGF-secreting cells, choline acetyltransferase-immunoreactive neurons within the striatum were hypertrophied relative to the contralateral side or control-grafted animals. Further, GFAP-hNGF-grafted rats displayed a robust sprouting of p75 neurotrophin receptor-positive fibers emanating from the underlying basal forebrain. These studies indicate that EGF-responsive stem cells which secrete hNGF under the direction of the GFAP promoter display in vitro and in vivo properties similar to that seen following other methods of NGF delivery and this source of cells may provide an excellent avenue for delivery of neurotrophins such as NGF to the central nervous system.
Assuntos
Transplante de Tecido Encefálico , Corpo Estriado/citologia , Fator de Crescimento Epidérmico/farmacologia , Transplante de Tecido Fetal , Proteína Glial Fibrilar Ácida/genética , Fatores de Crescimento Neural/genética , Transplante de Células-Tronco , Animais , Diferenciação Celular/efeitos dos fármacos , Meios de Cultivo Condicionados , Regulação da Expressão Gênica , Proteína Glial Fibrilar Ácida/biossíntese , Humanos , Masculino , Camundongos , Camundongos Transgênicos , Fatores de Crescimento Neural/biossíntese , Células PC12 , Ratos , Ratos Sprague-Dawley , Receptor de Fator de Crescimento Neural , Receptores de Fator de Crescimento Neural/análise , Proteínas Recombinantes de Fusão/biossíntese , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismoRESUMO
Tumor angiogenesis, the development of new blood vessels, is a highly regulated process that is controlled genetically by alterations in oncogene and tumor suppressor gene expression and physiologically by the tumor microenvironment. Previous studies indicate that the angiogenic switch in Ras-transformed cells may be physiologically promoted by the tumor microenvironment through the induction of the angiogenic mitogen, vascular endothelial growth factor (VEGF). In this report, we show Ras-transformed cells do not use the downstream effectors c-Raf-1 or mitogen activated protein kinases (MAPK) in signaling VEGF induction by hypoxia as overexpression of kinase-defective alleles of these genes does not inhibit VEGF induction under low oxygen conditions. In contrast to the c-Raf-1/MAP kinase pathway, hypoxia increases phosphatidylinositol 3-kinase (PI 3-kinase) activity in a Ras-dependent manner, and inhibition of PI 3-kinase activity genetically and pharmacologically results in inhibition of VEGF induction. We propose that hypoxia modulates VEGF induction in Ras-transformed cells through the activation of a stress inducible PI 3-kinase/Akt pathway and the hypoxia inducible factor-1 (HIF-1) transcriptional response element.
Assuntos
Transformação Celular Neoplásica , Proteínas de Ligação a DNA/fisiologia , Fatores de Crescimento Endotelial/fisiologia , Genes ras , Linfocinas/fisiologia , Neovascularização Patológica , Proteínas Nucleares/fisiologia , Fosfatidilinositol 3-Quinases/fisiologia , Transdução de Sinais , Células 3T3 , Animais , Hipóxia Celular , Regulação da Expressão Gênica , Fator 1 Induzível por Hipóxia , Subunidade alfa do Fator 1 Induzível por Hipóxia , Camundongos , Fatores de Transcrição/fisiologia , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio VascularRESUMO
The present study examined whether implants of epidermal growth factor (EGF)-responsive stems cells derived from transgenic mice in which the glial fibrillary acid protein (GFAP) promoter directs the expression of human nerve growth factor (hNGF) could prevent the degeneration of striatal neurons in a rodent model of Huntington's disease (HD). Rats received intrastriatal transplants of GFAP-hNGF stem cells or control stem cells followed 9 days later by an intrastriatal injection of quinolinic acid (QA). Nissl stains revealed large striatal lesions in rats receiving control grafts, which, on average, encompassed 12.78 mm3. The size of the lesion was significantly reduced (1.92 mm3) in rats receiving lesions and GFAP-hNGF transplants. Rats receiving QA lesions and GFAP-hNGF-secreting grafts stem cell grafts displayed a sparing of striatal neurons immunoreactive (ir) for glutamic acid decarboxylase, choline acetyltransferase, and neurons histochemically positive for nicotinamide adenosine diphosphate. Intrastriatal GFAP-hNGF-secreting implants also induced a robust sprouting of cholinergic fibers from subjacent basal forebrain neurons. The lesioned striatum in control-grafted animals displayed numerous p75 neurotrophin-ir (p75NTR) astrocytes, which enveloped host vasculature. In rats receiving GFAP-hNGF-secreting stem cell grafts, the astroglial staining pattern was absent. By using a mouse-specific probe, stem cells were identified in all animals. These data indicate that cellular delivery of hNGF by genetic modification of stem cells can prevent the degeneration of vulnerable striatal neural populations, including those destined to die in a rodent model of HD, and supports the emerging concept that this technology may be a valuable therapeutic strategy for patients suffering from this disease.