RESUMO
BACKGROUND: Information regarding the kinetics and longevity of acquired immunity in recovered COVID-19 patients requires thorough analysis and documentation. This is an update to an ongoing monocentric pilot observational study, that longitudinally analyzed the presence of antibodies after SARS-CoV-2 infection. STUDY DESIGN: Antibody titers against nucleocapsid protein (NCP) of SARS-CoV-2 analyzed at 8 months was followed by adoption of a more specific immunoassay, anti-Spike-Receptor binding domain IgG CLIA for analysis at 12 and 13 months post infection. METHODS: MAGLUMI® SARS-CoV-2 S-RBD IgG Chemiluminescence immunoassay (CLIA) was adopted for measurement of antibody titres at 12 and 13 months after SARS-CoV-2 infection. RESULTS: 97% (34 out of 35) patients resulted positive for anti-SARS-CoV-2 RBD IgG at 12 and 13 months. DISCUSSION AND CONCLUSIONS: In areas with vaccine and resource scarcity, vaccination could be prioritized for those individuals who have never been infected or for the ones who have recovered but show the absence of protective antibodies.
Assuntos
COVID-19 , SARS-CoV-2 , Anticorpos Neutralizantes , Anticorpos Antivirais , Humanos , Imunoglobulina G , Glicoproteína da Espícula de Coronavírus/química , Glicoproteína da Espícula de Coronavírus/metabolismoRESUMO
Introduction: A large amount of recent research has focused on the nature of immunity elicited by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, particularly its robustness and the duration of protection it offers. As a vaccine's efficacy relies on its ability to induce a protective immune response, these questions remain particularly pertinent. An improved understanding of the immunity offered by the antibodies developed against SARS-CoV-2 in recovered patients is critical for the development of diagnostic tests and vaccines. Methods: Our study aimed at the longitudinal analysis of antibody presence, persistence and its trend over eight months in a group of 30 COVID-19 recovered patients who tested positive by real-time quantitative PCR for SARS-CoV-2 in the period 1-30 March 2020. The subjects were divided into two groups based on disease severity: mild (n=17 subjects) and moderately-severe (n=13 subjects). The MAGLUMI 2019-nCoV lgM/lgG chemiluminescent analytical system (CLIA) assay was used to analyze these antibody titres. Results: IgG antibody persistency was demonstrated in 76.7 % of the subjects (23 out of 30) at eight months post-infection. For the moderately-severe group, the titre trends for both IgM and IgG changed in a statistically significant way throughout the time period with IgM below and IgG above the set cut-off. Conclusions: The results of this study highlight an important point in terms of the association between humoral immune response and disease severity. Patients who have experienced a relatively severe infection might develop a stronger immune response that could persist for a longer period.
Assuntos
COVID-19 , SARS-CoV-2 , Anticorpos Antivirais , Humanos , Imunoglobulina G , Reação em Cadeia da Polimerase em Tempo RealRESUMO
The tumor suppressor p53 is mainly involved in the transcriptional regulation of a large number of growth-arrest- and apoptosis-related genes. However, a clear understanding of which factor/s influences the choice between these two opposing p53-dependent outcomes remains largely elusive. We have previously described that in response to DNA damage, the RNA polymerase II-binding protein Che-1/AATF transcriptionally activates p53. Here, we show that Che-1 binds directly to p53. This interaction essentially occurs in the first hours of DNA damage, whereas it is lost when cells undergo apoptosis in response to posttranscriptional modifications. Moreover, Che-1 sits in a ternary complex with p53 and the oncosuppressor Brca1. Accordingly, our analysis of genome-wide chromatin occupancy by p53 revealed that p53/Che1 interaction results in preferential transactivation of growth arrest p53 target genes over its pro-apoptotic target genes. Notably, exposure of Che-1(+/-) mice to ionizing radiations resulted in enhanced apoptosis of thymocytes, compared with WT mice. These results confirm Che-1 as an important regulator of p53 activity and suggest Che-1 to be a promising yet attractive drug target for cancer therapy.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Apoptose/genética , Proteína BRCA1/metabolismo , Pontos de Checagem do Ciclo Celular/genética , Proteínas Repressoras/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Animais , Proteínas Reguladoras de Apoptose/genética , Linhagem Celular Tumoral , Dano ao DNA/genética , Reparo do DNA/genética , Ativação Enzimática/genética , Regulação da Expressão Gênica , Células HCT116 , Humanos , Células MCF-7 , Camundongos , Camundongos Transgênicos , Ligação Proteica/genética , Interferência de RNA , RNA Interferente Pequeno , Proteínas Repressoras/genética , Timócitos/patologia , Timócitos/efeitos da radiação , Ativação Transcricional/genética , Proteína Supressora de Tumor p53/genéticaRESUMO
Che-1/AATF is an RNA polymerase II-binding protein that is involved in the regulation of gene transcription, which undergoes stabilization and accumulation in response to DNA damage. We have previously demonstrated that following apoptotic induction, Che-1 protein levels are downregulated through its interaction with the E3 ligase HDM2, which leads to Che-1 degradation by ubiquitylation. This interaction is mediated by Pin1, which determines a phosphorylation-dependent conformational change. Here we demonstrate that HIPK2, a proapoptotic kinase, is involved in Che-1 degradation. HIPK2 interacts with Che-1 and, upon genotoxic stress, phosphorylates it at specific residues. This event strongly increases HDM2/Che-1 interaction and degradation of Che-1 protein via ubiquitin-dependent proteasomal system. In agreement with these findings, we found that HIPK2 depletion strongly decreases Che-1 ubiquitylation and degradation. Notably, Che-1 overexpression strongly counteracts HIPK2-induced apoptosis. Our results establish Che-1 as a new HIPK2 target and confirm its important role in the cellular response to DNA damage.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Apoptose , Proteínas de Transporte/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Repressoras/metabolismo , Proteínas Reguladoras de Apoptose/genética , Proteínas de Transporte/genética , Dano ao DNA , Humanos , Fosforilação , Ligação Proteica , Proteínas Serina-Treonina Quinases/genética , Proteólise , Proteínas Repressoras/genética , Ubiquitina/metabolismo , UbiquitinaçãoRESUMO
During space missions, astronauts work in a state of separation from their daily social environment and in physical confinement. It has been shown that confinement influences mood and brain cortical activity, but no data has been obtained with regard to its effect on the thyroid gland, the structure and function of which change during spaceflights. Here, we report the results of a study on the effects of confinement on mouse thyroid, which was implemented with the Mice Drawer System Facility maintained on the ground, a system used for spaceflight experiments. The results show that confinement changes the microscopic structure of the thyroid gland and that it exhibits symptoms similar to those that result from physiological and/or pathological hyperfunction. What is left unchanged, however, is the sphingomyelinase-thyrotropin receptor relationship, which is important for thyrotropin response with a consequential production of hormones that act on the metabolism of almost all tissues and reduces the production of calcitonin, a hormone involved in bone metabolism. During space missions, the overexpression of pleiotrophin, a widespread cytokine up-regulated after tissue injury that acts on bone remodeling, attenuates changes to the thyroid that are spaceflight-dependent; therefore we studied the thyroids of pleiotrophin-transgenic mice in the Mice Drawer System Facility. In confinement, pleiotrophin overexpression does not protect from the loss of calcitonin. The contribution of confinement to thyroid damage during spaceflights is discussed.
Assuntos
Calcitonina/metabolismo , Proteínas de Transporte/genética , Espaços Confinados , Citocinas/genética , Receptores da Tireotropina/metabolismo , Voo Espacial , Esfingomielina Fosfodiesterase/metabolismo , Glândula Tireoide/metabolismo , Tireotropina/metabolismo , Animais , Remodelação Óssea , Proteínas de Transporte/metabolismo , Citocinas/metabolismo , Camundongos , Camundongos Transgênicos , Glândula Tireoide/patologiaRESUMO
X-linked inhibitor of apoptosis protein (XIAP) is a member of the inhibitor of apoptosis proteins family that selectively binds and inhibits caspase-3, -7 and -9. As such, XIAP is an extremely potent suppressor of apoptosis and an attractive target for cancer treatment. Che-1 is an antiapoptotic agent involved in the control of gene transcription and cell proliferation. Recently, we showed that the checkpoint kinases ATM/ATR and checkpoint kinase 2 physically and functionally interact with Che-1 and promote its phosphorylation and accumulation in response to DNA damage. These Che-1 modifications induce transcription of p53, and Che-1 depletion strongly sensitizes tumor cells to anticancer drugs. Here we show that Che-1 activates XIAP expression in response to DNA damage. This effect is mediated by Che-1 phosphorylation and requires NF-kappaB. Notably, we found that XIAP expression is necessary for antiapoptotic activity of Che-1 and that in vivo downregulation of Che-1 by small interference RNA strongly enhanced the cytotoxicity of anticancer drugs.
Assuntos
Proteínas Reguladoras de Apoptose/fisiologia , Apoptose , Dano ao DNA , Proteínas Repressoras/fisiologia , Fatores de Transcrição/fisiologia , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/biossíntese , Animais , Proteínas Reguladoras de Apoptose/antagonistas & inibidores , Proteínas Reguladoras de Apoptose/genética , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos , Humanos , Masculino , Camundongos , Camundongos Nus , NF-kappa B/metabolismo , Células NIH 3T3 , Interferência de RNA , Proteínas Repressoras/antagonistas & inibidores , Proteínas Repressoras/genética , Fatores de Transcrição/antagonistas & inibidores , Fatores de Transcrição/genética , Ativação Transcricional , Regulação para Cima , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/genéticaRESUMO
Sixty, relapsing remitting (RR) multiple sclerosis (MS) patients, who underwent treatment with glatiramer acetate (GA), interferon (IFN)-beta 1a, and immunoglobulins (Igs) (20 per treatment group), were assessed for levels of brain-derived neurotrophic factor (BDNF) in the supernatants of unstimulated and stimulated peripheral blood mononuclear cells (PBMCs) in the first year of treatment. Phytohemagglutinin (PHA), anti-OKT3 antibody, myelin basic protein (MPB) and GA were used as stimuli. Cytokine responses by ELISPOT and lymphoproliferative responses were also assessed. The GA-treated MS patient group showed a progressive increase in BDNF levels, from baseline to month three; thereafter, the levels remained stable and significantly greater compared with baseline and controls (ANOVA=P<0.001). IFN-beta 1a had no effect on BDNF production, whereas Igs induced a slight decrease (ANOVA=P<0.04). ELISPOT analysis revealed a significant decrease of IFN-gamma, an increase of interleukin (IL)-4 and IL-5 in GA-treated MS patients, and an increase of IL-10 in patients treated with IFN-beta 1a and GA. No significant correlation was found between BDNF secretion in the supernatants of PBMCs and cytokine response, lesional load, and measures of atrophy. Increased BDNF production related to GA treatment can have implications for understanding the mechanism of action of this immunomodulatory agent, in light of evidence suggesting its effects in promoting neuroprotective immunity in MS patients; however, a clinically measurable effect, especially in terms of an impact on actual disease progression, remains to be established.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/sangue , Imunoglobulinas/uso terapêutico , Interferon beta/uso terapêutico , Leucócitos Mononucleares/fisiologia , Esclerose Múltipla Recidivante-Remitente/sangue , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico , Peptídeos/uso terapêutico , Adjuvantes Imunológicos/uso terapêutico , Adulto , Encéfalo/anatomia & histologia , Encéfalo/patologia , Feminino , Seguimentos , Acetato de Glatiramer , Humanos , Interferon beta-1a , Leucócitos Mononucleares/efeitos dos fármacos , Ativação Linfocitária , Contagem de Linfócitos , Imageamento por Ressonância Magnética , Masculino , Fatores de TempoRESUMO
This study investigated nuclear factor-kappa B (NF-kappaB) activity by electrophoresis mobility gel shift assay and IkappaBalpha expression by Western blot analysis in monocytes obtained from serial samples of internal jugular venous blood taken from seven migraine patients without aura during attacks. Inducible nitric oxide synthase (iNOS) expression was also assessed by reverse transcription-polymerase chain reaction. An increase in NF-kappaB activity peaked 2 h after attack onset. This was accompanied by a transient reduction in IkappaBalpha expression. Up-regulation of iNOS was evident at 4 h, maintained at 6 h and reduced at the end of the attack. These findings substantiate the hypothesis of transitory delayed inflammation, as suggested by the animal model, and suggest the possibility of using therapeutic approaches to target NF-kappaB transcription in the treatment of migraine.
Assuntos
Veias Jugulares/metabolismo , Enxaqueca sem Aura/sangue , Enxaqueca sem Aura/patologia , Monócitos/metabolismo , NF-kappa B/sangue , Óxido Nítrico Sintase Tipo II/sangue , Adulto , Ativação Enzimática , Feminino , Expressão Gênica , Humanos , MasculinoRESUMO
The aim of the present study was to verify cerebrospinal fluid (CSF) levels of glial cell line-derived neurotrophic factor (GDNF) and somatostatin, both measured by sensitive immunoassay, in: 16 chronic migraine (CM) patients, 15 patients with an antecedent history of migraine without aura diagnosed as having probable chronic migraine (PCM) and probable analgesic-abuse headache (PAAH), 20 patients affected by primary fibromyalgia syndrome (PFMS), and 20 control subjects. Significantly lower levels of GDNF and somatostatin were found in the CSF of both CM and PCM + PAAH patients compared with controls (GDNF =P < 0.001, P < 0.002; somatostatin = P < 0.002, P < 0.0003), without significant difference between the two groups. PFMS patients, with and without analgesic abuse, also had significantly lower levels of both somatostatin and GDNF (P < 0.0002, P < 0.001), which did not differ from those of CM and PCM + PAAH patients. A significant positive correlation emerged between CSF values of GDNF and those of somatostatin in CM (r = 0.70, P < 0.02), PCM + PAAH (r = 0.78, P < 0.004), and PFMS patients (r = 0.68, P < 0.008). Based on experimental findings, it can be postulated that reduced CSF levels of GDNF and somatostatin in both CM and PCM + PAAH patients can contribute to sustained central sensitization underlying chronic head pain. The abuse of simple or combination analgesics does not seem to influence the biochemical changes investigated, which appear to be more strictly related to the chronic pain state, as demonstrated also for fibromyalgia.
Assuntos
Analgésicos , Fibromialgia/líquido cefalorraquidiano , Fator Neurotrófico Derivado de Linhagem de Célula Glial/líquido cefalorraquidiano , Transtornos de Enxaqueca/líquido cefalorraquidiano , Somatostatina/líquido cefalorraquidiano , Transtornos Relacionados ao Uso de Substâncias/líquido cefalorraquidiano , Adulto , Biomarcadores/líquido cefalorraquidiano , Doença Crônica , Comorbidade , Feminino , Fibromialgia/epidemiologia , Humanos , Itália/epidemiologia , Masculino , Transtornos de Enxaqueca/epidemiologia , Transtornos Relacionados ao Uso de Substâncias/epidemiologiaRESUMO
The present study was aimed at verifying the clinical characteristics of a typical attack in 20 migraine patients, 10 responders and 10 non-responders to rizatriptan, and at investigating any differences in the levels of neuropeptides of the trigeminovascular or parasympathetic systems [calcitonin gene-related peptide (CGRP), neurokinin A (NKA) and vasoactive intestinal peptide (VIP) measured by radioimmunoassay methods in external jugular blood] between responders and non-responders. In all responders to rizatriptan, pain was unilateral, severe, and pulsating, and in five of them at least one sign suggestive of parasympathetic system activation was recorded. Five patients who were non-responders to rizatriptan referred bilateral and non-pulsating pain, even though severe in most of them. CGRP and NKA levels measured before rizatriptan administration were significantly higher in responders than in non-responders (P < 0.0001 and P < 0.002, respectively). In the five patients with autonomic signs among rizatriptan responders, detectable VIP levels were found at baseline. One hour after rizatriptan administration, a decrease in CGRP and NKA levels was evident in the external jugular venous blood of rizatriptan responders, and this corresponded to a significant pain relief and alleviation of accompanying symptoms. VIP levels were also significantly reduced at the same time in the five patients with autonomic signs. After rizatriptan administration, CGRP and NKA levels in non-responder patients showed less significant variations at all time points after rizatriptan administration compared with rizatriptan responders. The present study, although carried out on a limited number of patients, supports recent clinical evidence of increased trigeminal activation associated with a better triptan response in migraine patients accompanied by parasympathetic activation in a subgroup of patients with autonomic signs. In contrast, the poor response seems to be correlated with a lesser degree of trigeminal activation, lower variations of trigeminal neuropeptides after triptan administration, and no evidence of parasympathetic activation at baseline.
Assuntos
Transtornos de Enxaqueca/tratamento farmacológico , Transtornos de Enxaqueca/fisiopatologia , Agonistas do Receptor de Serotonina/uso terapêutico , Triazóis/uso terapêutico , Triptaminas/uso terapêutico , Peptídeo Relacionado com Gene de Calcitonina/sangue , Peptídeo Relacionado com Gene de Calcitonina/efeitos dos fármacos , Resistência a Medicamentos , Humanos , Técnicas Imunoenzimáticas , Transtornos de Enxaqueca/sangue , Neurocinina A/sangue , Neurocinina A/efeitos dos fármacos , Peptídeo Intestinal Vasoativo/sangue , Peptídeo Intestinal Vasoativo/efeitos dos fármacosRESUMO
Thrombolytic therapy not always improves clinical outcome in ischemic stroke patients. This could cause lymphomonocyte accumulation in the infarcted brain area. These produce an excessive amount of proinflammatory cytokines, such as IL-1 beta, IL-6 and TNF-alfa. The aim of our study was to determine ILs levels in fibrinolytic therapy treated patients, compared with healthy controls and to evaluate if the varying levels can predictors of neurological outcome. Eighteen patients underwent thrombolytic treatment with t-PA within 3 h. Plasma levels of IL-1 beta, IL-6, TNF-alfa and IL-10 were determined by ELISA method before and within 24 h after t-PA infusion and compared with controls. Significantly higher levels of IL-1 beta and Il-6 emerged in stroke patients before treatment compared with the control group (P < 0.05 and 0.04, respectively). Slightly higher plasma levels of TNF-alfa and lower plasma levels of IL-10 were also found at base line in stroke patients. After thrombolytic treatment no significant variations were observed in the levels of TNF-alfa and IL-6, whereas a trend toward lower values for IL-1 beta and higher levels for IL-10 was observed. Positive correlations among the values of IL-6, TNF-alfa and National Institute of Health Stroke Scale (NIHSS) at discharges were observed. A similar correlation with modified Rankin scale score at 3 month was found. Pre-treatment cytokine status seems to influence pre-and long-term clinical outcome. Therefore an investigation into the possible predictor of cytokines seem worthy.
Assuntos
Citocinas/análise , Fibrinolíticos/uso terapêutico , Acidente Vascular Cerebral/tratamento farmacológico , Terapia Trombolítica , Ativador de Plasminogênio Tecidual/uso terapêutico , Idoso , Idoso de 80 Anos ou mais , Análise de Variância , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Fatores de Risco , Estatísticas não Paramétricas , Acidente Vascular Cerebral/diagnóstico , Acidente Vascular Cerebral/metabolismo , Terapia Trombolítica/métodos , Fatores de Tempo , Resultado do TratamentoRESUMO
The role of hyperhomocysteinemia as independent risk factor for stroke needs to be confirmed. The aims of our study were to assess (i) the association between risk of stroke and increasing values of plasma homocysteine and (ii) the interaction between mild hyperhomocysteinemia and conventional vascular risk factors. We studied 161 consecutive patients with first-ever ischemic stroke classified using TOAST criteria and 152 neurologically healthy controls. Homocysteine was measured using high performance liquid chromatography (HPLC). Homocysteinemia was elevated in all stroke subtypes: 13.0+/-2.5 micromol/l in patients with cardioembolic disease, 13.9+/-5.4 micromol/l in those with small vessel diseases, 15.5+/-6.8 micromol/l in cases of undetermined stroke, and 17.8+/-13.5 micromol/l in patients with large vessel disease. Mean homocysteinemia was 8.10 micromol/l (SD=2.5) in controls. The logistic regression analysis showed that important independent risk factors for ischemic stroke were hypertension (p<0.0001; OR= 3.205; 95% CI, 1.788-5.742), hyperhomocysteinemia (p<0.0001; OR=1.425; 95% CI, 1.300-1562) and hyperlipidemia (p=0.018; OR=2.243; 95% CI, 1.147-4.385). Hyperhomocyst(e)inemia is an independent risk factor for all stroke subtypes and should be routinely measured and treated in stroke patients.
Assuntos
Homocisteína/sangue , Hiper-Homocisteinemia/complicações , Acidente Vascular Cerebral/sangue , Acidente Vascular Cerebral/etiologia , Idoso , Cromatografia Líquida de Alta Pressão , Complicações do Diabetes , Feminino , Humanos , Hiperlipidemias/complicações , Hipertensão/complicações , Masculino , Fatores de Risco , Fumar/efeitos adversosRESUMO
BACKGROUND & AIMS: Carboxyethyl-hydroxychromans (CEHC) are hydrosoluble vitamin E metabolites excreted through the renal filter. In this study we investigated the effect of the kidney damage on the blood levels of CEHC. METHODS: Plasma levels of alpha-CEHC, gamma-CEHC and their precursors (namely, alpha-tocopherol and gamma-tocopherol) were measured by HPLC with electrochemical detection in chronic (CRF) and end-stage renal failure patients on regular hemodialysis (HD) before and after dialysis. CRF patients (n = 26) were divided into three subgroups with different extent of kidney damage as measured by the intervals of creatinine clearance (CrCl, in ml/min): (a) 2-10, (b) 10-20, and (c) 20-45. HD patients (n = 8) did not show residual renal function. In all the subjects the intake of vitamin E (as alpha-tocopherol) was assessed using a food frequency questionnaire. In the HD group, the plasma concentrations of ascorbic and uric acid (AA and UA, respectively), total thiols, the total antioxidant status (TAS) and reactive carbonyls were also measured. RESULTS: The progressive deterioration of the kidney function in the different groups of patients produced an exponential increase of both alpha-CEHC and gamma-CEHC in plasma. Compared with healthy controls (alpha-CEHC = 20.1+/-13.4 and gamma-CEHC = 230.6+/-83.0 nmol/l) the levels of CEHC approximately doubled in patients with CrCl < or = 20ml/min (42.4+/-20.2 and 424.5.5+/-174.4; P <0.05 or higher in both) and reached a 3-fold maximum increase in HD patients (77.3+/-45.7 and 636.6+/-219.3). The hemodialysis provided a significant, but only a transient, correction of CEHC accumulation (44.8+/-23.5, 364.2+/-189.9). The HD patients showed lower intake and levels of vitamin E (alpha-tocopherol = 5.1+/-1.0 and gamma-tocopherol =0.32+/-0.11 micromol/mmol cholesterol; P <0.05) compared to healthy controls (5.8+/-0.8 and 0.43+/-0.14), but in the CRF patients tocopherol levels were normal or only slightly decreased even though approximately half of the subject had lowered vitamin E intake. When the entire patient population was considered, the blood concentrations of parental tocopherols and CEHC did not correlate. The HD patients before dialysis showed a marked decrease of TAS/UA, AA and thiols levels, while UA and free carbonyls significantly increased. After dialysis, the depletion of AA and thiols further worsened and also UA and TAS/UA decreased, but free carbonyls slightly increased. CONCLUSIONS: The results other than to confirm the key importance of the renal route for the excretion of CEHC, demonstrate that CEHC cannot be reliably used to investigate vitamin E biokinetics and transformation without a careful examination of the renal function. CEHC accumulation does not seem to influence the antioxidant status in the plasma of HD patients. Further studies are requested to establish whether such an increase in blood CEHC concentrations might be harmful or could contribute to the biological functions of the vitamin E in uremia and dialysis patients.
Assuntos
Injúria Renal Aguda/sangue , Falência Renal Crônica/sangue , Vitamina E/sangue , Injúria Renal Aguda/fisiopatologia , Injúria Renal Aguda/terapia , Adulto , Idoso , Antioxidantes/análise , Cromanos/sangue , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Rim/fisiopatologia , Falência Renal Crônica/fisiopatologia , Falência Renal Crônica/terapia , Masculino , Pessoa de Meia-Idade , Valores de Referência , Diálise Renal , Vitamina E/administração & dosagem , alfa-Tocoferol/sangue , gama-Tocoferol/sangueRESUMO
BACKGROUND: The complication rate of central venous totally implantable access ports (TIAP), used for high-dose chemotherapy with autologous stem cell transplantation support, has not been fully investigated to date, due to the almost exclusive use of externalised, tunnelled devices in this clinical setting. PATIENTS AND METHODS: During a 66-month period (from 1 January 1997 to 30 June 2002), 376 patients suffering from breast cancer, ovarian cancer, lymphoma or multiple myeloma were treated with high-dose chemotherapy and autologous stem cell transplantation at the European Institute of Oncology (Milan, Italy). A single type of port was used, constructed from titanium and silicone rubber, connected to a 7.8 F polyurethane catheter (Port-A-Cath; SIMS Deltec, Inc., St Paul, MN, USA) inserted into the subclavian vein. They were followed prospectively for device-related complications until the device was removed, the patient died or the study was closed (30 June 2002). RESULTS: No TIAP-related deaths were observed in this series. Seven pneumothoraxes (1.8%) occurred as a complication of TIAP placement, one patient only (0.2%) requiring a tube thoracostomy. Port pocket infection occurred twice in this series (0.53%, 0.01 episodes/1000 days of use), whereas three patients suffered from port-related bacteraemia (0.8%, 0.016/1000 days of use). Infections were successfully treated with antibiotics; all three cases had the ports removed at programme completion. Four cases of deep vein thrombosis were detected (1.06%, 0.022/1000 days of use); low molecular weight heparin was given, followed by oral anticoagulants. Finally, one case of extravasation occurred (0.26%, 0.005/1000 days of use), requiring port removal and local medical therapy. CONCLUSIONS: The use of TIAPs has resulted in a safe and effective option for high-dose chemotherapy deliverance and stem cell transplantation, in spite of inducing severe neutropenia and increasing the risk of sepsis in this category of oncology patient.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Cateterismo Venoso Central/efeitos adversos , Transplante de Células-Tronco de Sangue Periférico , Pneumotórax/etiologia , Adulto , Neoplasias da Mama/tratamento farmacológico , Feminino , Humanos , Infecções/etiologia , Linfoma/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/tratamento farmacológico , Neoplasias Ovarianas/tratamento farmacológico , Estudos Prospectivos , Fatores de RiscoRESUMO
A central sensitization has been advocated to explain chronic daily headache (CDH) due to sustained peripheral sensitization of allogenic structures responsible for sustained trigeminovascular system activation. Several mechanisms have been suggested to underlie central sensitization, but have been poorly investigated in CDH. They involve N-methyl-D-aspartate (NMDA) receptor activation and nitric oxide (NO) production and supersensitivity and increased and maintained production of sensory neuropeptides. The present study supports the above pathogenic mechanisms demonstrating a significant increase in glutamate and nitrite levels in the CSF of CDH patients, without a significant difference between patients without and those with analgesic overuse headache (P < 0.0001 and P < 0.002). The increase in CSF nitrites was accompanied by a significant rise in the CSF values of cyclic guanosine monophosphate (cGMP) in patients in comparison with controls (P < 0.0001). A statistically significant correlation emerged between visual analogic scale (VAS) values and glutamate, nitrites and cGMP. Although substance P (SP) and calcitonin gene-related peptide (CGRP), and to a lesser extent neurokinin A, were significantly increased in CSF compared with control subjects, their values did not correlate with glutamate, nitrites and cGMP levels in CSF in the patient group. The present study confirms the involvement of glutamate-NO-cGMP-mediated events underlying chronic head pain that could be the target of a new therapeutic approach which should be investigated.
Assuntos
Ácido Glutâmico/líquido cefalorraquidiano , Transtornos da Cefaleia/líquido cefalorraquidiano , Óxido Nítrico/líquido cefalorraquidiano , Adulto , Análise de Variância , Intervalos de Confiança , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The aim of this paper was to evaluate the role of bcl-2 in the susceptibility of the MCF7 ADR human breast carcinoma line overexpressing the P-170 glycoprotein (P-170) to various drugs. The sensitivity to four multidrug resistance (MDR)-related drugs (doxorubicin (ADR), vincristine (VCR), vinblastine (VBL), actinomycin D (ACTD)) and three MDR-non-related drugs (cisplatin (DDP), bischloroethylnitrosourea (BCNU), 5-fluorouracil (5-FU)) was evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay in three bcl-2-overexpressing clones obtained from the MCF7 ADR line. We found that the bcl-2-overexpressing clones show increased resistance to DDP and BCNU, while no difference to 5-FU were observed between the control cells and bcl-2 transfectants. Surprisingly, bcl-2-overexpressing clones displayed an increased sensitivity compared with the control cells to the MDR-related drugs ADR, VCR, VBL and ACTD. Focusing on DDP and ADR, we found that the increased resistance of the bcl-2 transfectants to DDP was correlated to their ability to prevent apoptosis, while the enhanced sensitivity to ADR was associated with an increased ADR accumulation and a decreased ADR efflux. Moreover, while bcl-2 overexpression does not induce changes in P-170 glycoprotein expression, it did induce a reduction of the adenosine triphosphate (ATP) levels and basal protein kinase C (PKC) activity, both of which have a crucial role in the regulation of the MDR phenotype. In conclusion, the effect of bcl-2 on antineoplastic sensitivity observed in this study underscores the idea that bcl-2 may have distinct biological effects depending on the anticancer drug used.
Assuntos
Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Genes bcl-2/fisiologia , Trifosfato de Adenosina/análise , Análise de Variância , Apoptose/efeitos dos fármacos , Western Blotting , Neoplasias da Mama/genética , Resistência a Múltiplos Medicamentos/genética , Resistencia a Medicamentos Antineoplásicos/genética , Feminino , Humanos , Proteína Quinase C/análise , Células Tumorais CultivadasRESUMO
Nitric oxide (NO) can modulate red blood cell (RBC) glycolysis by translocation of the enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPD) (EC 1.2.1.12) from the cytosolic domain of the membrane protein band 3 (cdb3) in the cytosol. In this study we have investigated which NO-reactive thiols might be influencing GAPD translocation and the specific role of glutathione. Two highly reactive Cys residues were identified by transnitrosylation with nitrosoglutathione (GSNO) of cdb3 and GAPD (K(2) = 73.7 and 101.5 M(-1) s(-1), respectively). The Cys 149 located in the catalytic site of GAPD is exclusively involved in the GSNO-induced nitrosylation. Reassociation experiments carried out at equilibrium with preparations of RBC membranes and GAPD revealed that different NO donors may form -SNO on, and decrease the affinity between, GAPD and cdb3. In intact RBC, the NO donors 3-morpholinosydnonimine (SIN-1) and peroxynitrite (ONOO(-)) significantly increased GAPD activity in the cytosol, glycolysis measured as lactate production, and energy charge levels. Our data suggest that ONOO(-) is the main NO derivative able to cross the RBC membrane, leading to GAPD translocation and -SNO formation. In cell-free experiments and intact RBC, diamide (a thiol oxidant able to inhibit GAPD activity) was observed to reverse the effect of SIN-1 on GAPD translocation. The results demonstrate that cdb3 and GAPD contain reactive thiols that can be transnitrosylated mainly by means of GSNO; these can ultimately influence GAPD translocation/activity and the glycolytic flux.
Assuntos
Proteína 1 de Troca de Ânion do Eritrócito/metabolismo , Eritrócitos/metabolismo , Glutationa/metabolismo , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico/metabolismo , Compostos de Sulfidrila/metabolismo , Cisteína/metabolismo , Citosol/metabolismo , Eritrócitos/efeitos dos fármacos , Glicólise/efeitos dos fármacos , Glicólise/fisiologia , Humanos , CinéticaRESUMO
The aim of the present study was to verify the expression of tumour necrosis factor (TNF)-alpha mRNA by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in unstimulated peripheral blood mononuclear cells (MNCs) of 15 relapsing-remitting multiple sclerosis (MS) patients who underwent treatment with IFN-beta 1a (6 millions of international units (MIU) i.m. once a week) and in 15 untreated MS patients matched for age and expanded disability status score (EDSS). At the same time the expression of TNF-alpha mRNA was assessed in 10 healthy age-matched control subjects. All MS patients were assessed at the basal time and after 6 months. At the basal time, the band of TNF-alpha mRNA was detectable in 12 out of the 15 untreated patients and in 13 out of the 15 patients who underwent IFN-beta 1a treatment. The higher TNF-alpha mRNA was evident in patients with gadolinium-enhancing lesions. At the 6-month follow-up, 13 out of the 15 untreated patients still had detectable values of TNF-alpha mRNA and no significant difference emerged when compared with basal time. On the contrary, the expression of TNF-alpha mRNA was absent at the same time in nine out of the 15 patients treated with IFN-beta 1a. A longitudinal analysis carried out monthly in eight MS patients (four untreated and four treated) revealed a transient increase in TNF-alpha mRNA expression in MNCs of all four treated patients in the first 3 months, supporting previous findings of an early immunoenhancing effect of IFN-beta 1a. This early activation is followed by an inhibitory effect of IFN-beta 1a on TNF-alpha mRNA expression in about 2/3 of treated MS patients when assessed at 6 months. Further long-term studies are needed to confirm this immunomodulatory effect of IFN-beta 1a not only on TNF-alpha but also on other cytokines of Th(1)and Th(2)types.