RESUMO
Oligodendrocyte gene expression is downregulated in stress-related neuropsychiatric disorders, including depression. In mice, chronic social stress (CSS) leads to depression-relevant changes in brain and emotional behavior, and the present study shows the involvement of oligodendrocytes in this model. In C57BL/6 (BL/6) mice, RNA-sequencing (RNA-Seq) was conducted with prefrontal cortex, amygdala and hippocampus from CSS and controls; a gene enrichment database for neurons, astrocytes and oligodendrocytes was used to identify cell origin of deregulated genes, and cell deconvolution was applied. To assess the potential causal contribution of reduced oligodendrocyte gene expression to CSS effects, mice heterozygous for the oligodendrocyte gene cyclic nucleotide phosphodiesterase (Cnp1) on a BL/6 background were studied; a 2 genotype (wildtype, Cnp1+/- ) × 2 environment (control, CSS) design was used to investigate effects on emotional behavior and amygdala microglia. In BL/6 mice, in prefrontal cortex and amygdala tissue comprising gray and white matter, CSS downregulated expression of multiple oligodendroycte genes encoding myelin and myelin-axon-integrity proteins, and cell deconvolution identified a lower proportion of oligodendrocytes in amygdala. Quantification of oligodendrocyte proteins in amygdala gray matter did not yield evidence for reduced translation, suggesting that CSS impacts primarily on white matter oligodendrocytes or the myelin transcriptome. In Cnp1 mice, social interaction was reduced by CSS in Cnp1+/- mice specifically; using ionized calcium-binding adaptor molecule 1 (IBA1) expression, microglia activity was increased additively by Cnp1+/- and CSS in amygdala gray and white matter. This study provides back-translational evidence that oligodendrocyte changes are relevant to the pathophysiology and potentially the treatment of stress-related neuropsychiatric disorders.
Assuntos
Oligodendroglia/metabolismo , Comportamento Social , Estresse Psicológico/genética , Transcriptoma , Tonsila do Cerebelo/metabolismo , Animais , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 1/genética , Nucleotídeo Cíclico Fosfodiesterase do Tipo 1/metabolismo , Hipocampo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Córtex Pré-Frontal/metabolismo , Estresse Psicológico/metabolismoRESUMO
OBJECTIVES: Vocal fold paralysis can have an important impact on a patient's quality of life. The goal of this study was to compare, in terms of vocal improvement and motility recovery, the post-vocal treatment results of our patients with unilateral vocal fold paralysis (UVFP) when treatment was started early (within 4 weeks from injury) versus intermediate (from 4 to 8 weeks) or delayed (at least 8 weeks after injury) treatment. STUDY DESIGN: An 11-year retrospective study of patients with UVFP who underwent multidimensional diagnostic-therapeutic assessment. METHODS: In total, 171 patients with UVFP were included in our study, divided into three groups who underwent early (first group), intermediate (second group), or delayed (third group) voice treatment. All patients underwent voice therapy based on forcible exercises supplemented by manipulations and maneuvers. RESULTS: Of the 171 patients with UVFP, 106 (62%) recovered vocal fold motility. Of these 106 patients, 51/78 (65%) were in the first group, 30/49 (61%) in the second group, and 25/44 (56%) in the third group. A significant (P < 0.0001) reduction in fundamental frequency (Fo) was present in the first group with a manifest improvement in the mean values of Jitter (Jitt%; P = 0.001), Shimmer (Shim%; P < 0.0001), and noise-to-harmonic ratio (NHR; P < 0.0001). A significant (P < 0.0001) reduction in Fo was found in the second group with a manifest improvement in Jitt% (P < 0.001), Shim% (P < 0.0001), and NHR (P < 0.0001). For the third group, no values were statistically significant apart from the improvement in NHR (P < 0.001). CONCLUSIONS: This study confirms the importance of early rehabilitation underlining the non-functional vocal recovery in patients who started treatment later than 8 weeks after injury.
Assuntos
Paralisia das Pregas Vocais/terapia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
To analyse the complications related to pexy, the main clinical manifestations that may raise suspicions of a pexy line rupture/detachment, the most suitable diagnostic technique and the optimum treatment to resolve this complication. This is a retrospective chart review in tertiary university referral centre. Medical charts of patients with oncological laryngeal pathologies admitted to the Otolaryngology Department of the University Hospital of Modena between May 2003 and March 2012 were analysed. Ten patients with rupture of the pexy were identified and included in the present study. The clinical manifestations were dysphagia, alteration of sensitivity of hypopharyngeallaryngeal structures, fever, infection and diastasis of surgical wounds, bleeding, dysphonia and aspiration pneumonia. Rupture of the pexy was diagnosed through endoscopic evaluations, radiological techniques or directly in the operating room during revision surgery of the earlier operation. Surgical treatments, coupled with effective swallowing rehabilitation, allowed progressive functional recovery. Patients were hospitalised until recovery of laryngeal functions was complete. In conclusion, pexy line rupture is one of the complications in the post-operative period of partial laryngectomies. Certain clinical manifestations may indicate this complication, helping the surgeon to establish an early diagnosis and administer prompt treatment.
Assuntos
Neoplasias Laríngeas/cirurgia , Laringectomia/efeitos adversos , Laringectomia/métodos , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
The objective of this study was to document functional results and to compare objective and subjective voice measures after endoscopic laryngoplasty with injection of polydimethylsiloxane (PDMS) for the treatment of unilateral vocal fold paralysis, and to verify PDMS biocompatibility in vocal fold. The design used was a longitudinal prospective study. Fifteen patients with unilateral vocal fold paralysis underwent endoscopic injection of PDMS in general anesthesia. Accurate voice evaluation protocol (acoustic and aerodynamics analyses, GIRBAS [Grade, Instability, Roughness, Breathiness, Asthenia, and Strain] scale, videostrobolaryngoscopy, and Voice Handicap Index test) before, after surgery, and at follow-up time was performed. The median follow-up was 21.7 months (range, 6-35). Data obtained were statistically significant. All acoustic, aerodynamics, perceptive, and subjective evaluations showed a significant improvement. No complications due to PDMS were reported. Functional results were found comparable to framework surgery. Endoscopic injection laryngoplasty with PDMS is a safe and long-term option for treatment of unilateral vocal fold paralysis.
Assuntos
Materiais Biocompatíveis/uso terapêutico , Dimetilpolisiloxanos/uso terapêutico , Procedimentos Cirúrgicos Otorrinolaringológicos/métodos , Paralisia das Pregas Vocais/tratamento farmacológico , Paralisia das Pregas Vocais/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Materiais Biocompatíveis/administração & dosagem , Dimetilpolisiloxanos/administração & dosagem , Feminino , Seguimentos , Lateralidade Funcional , Humanos , Masculino , Pessoa de Meia-Idade , Fonação/efeitos dos fármacos , Fonação/fisiologia , Estudos Prospectivos , Fatores de Tempo , Resultado do Tratamento , Paralisia das Pregas Vocais/fisiopatologia , Adulto JovemRESUMO
AIM: Descending necrotizing mediastinitis (DNM) is an unusual and severe disease with a high mortality rate. Surgical management remains controversial. Our investigations reviews the most effective surgical treatment in the management of this rare pathology. METHODS: Seven patients with DNM and treated over a 20-year period are reported. All patients were evaluated according to the classification suggested by Endo et al. of the degree of mediastinal diffusion, based on CT scan findings. Five patients underwent combined cervical drainage and thoracotomy, 2 patients were treated with cervical drainage alone. RESULTS: The outcome was favorable in 5 patients, 4 treated with a combined cervical and thoracic approach and 1 with a cervical approach alone. Two patients that underwent a combinated cervical and thoracic approach alone, died of septic shock. Overall mortality rate was 28.5%. CONCLUSION: Early diagnosis and early, aggressive surgical treatment are required to improve the poor prognosis of DNM. Although a unique surgical management is still not completely accepted, we state, in agreement with other authors, a wide approach consisting of a cervical drainage and mediastinotomy in case of upper mediastinitis and a combined cervical and thoracic approach in case of lower mediastinitis. In the course of thoracotomy a wide excision of necrotic and particularly fat mediastinal tissue is needed, to avoid a recurrent infection. A continuous cervico-mediastinal irrigation system is suggested during the postoperative period.
Assuntos
Mediastinite/diagnóstico por imagem , Mediastinite/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Criança , Drenagem/métodos , Feminino , Humanos , Masculino , Mediastinite/patologia , Pessoa de Meia-Idade , Necrose , Prognóstico , Doenças Estomatognáticas/complicações , Doenças Estomatognáticas/diagnóstico , Doenças Estomatognáticas/terapia , Toracotomia/métodos , Tomografia Computadorizada por Raios XRESUMO
OBJECTIVE: The study was part of a randomized open-label clinical trial designed to evaluate the effects of intra-articular injections of hyaluronan (Hyalgan) (HY) in osteoarthritis (OA) of the human knee. Data were compared with those obtained after treatment with methylprednisolone acetate (Depomedrol) (MP). METHODS: Synovial membranes from patients with OA of the knee, primary or secondary to a traumatic event and classified according to the American College of Rheumatology criteria, were examined by arthroscopy and by light and electron microscopy before and 6 months after local injection of HY (2 ml of 500-730 000 MW hyaluronan, 10 mg/ml in saline, one injection per week for 5 weeks) or MP (1 ml of methylprednisolone acetate, 40 mg/ml, one injection per week for 3 weeks). RESULTS: Arthroscopy revealed a significant decrease in inflammatory score after both treatments. Histology showed that HY treatment was effective (P< or =0.05) in reducing the number and aggregation of lining synoviocytes, as well as the number and calibre of the vessels. MP treatment significantly reduced the number of mast cells in primary OA. Both treatments tended to decrease the number of hypertrophic and to increase the number of fibroblast-like lining cells, to decrease the numbers of macrophages, lymphocytes, mast cells and adipocytes, and to decrease oedema, especially in primary OA, and to increase the number of fibroblasts and the amount of collagen. These phenomena were evident throughout the thickness of the synovial tissue. CONCLUSION: At least in the medium term, both HY and MP modified a number of structural variables of the synovial membrane of the osteoarthritic human knee towards the appearance of that of normal synovium. The effect was more evident in primary OA than in OA secondary to a traumatic event. This is the first evidence that local hyaluronan injections modify the structural organization of the human knee synovium in OA.
Assuntos
Adjuvantes Imunológicos/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Ácido Hialurônico/uso terapêutico , Articulação do Joelho/patologia , Metilprednisolona/análogos & derivados , Metilprednisolona/uso terapêutico , Osteoartrite/patologia , Membrana Sinovial/patologia , Adulto , Biópsia , Feminino , Humanos , Injeções Intra-Articulares , Masculino , Acetato de Metilprednisolona , Pessoa de Meia-Idade , Osteoartrite/tratamento farmacológicoRESUMO
In eukaryotic cells, efficient translation of most cellular mRNAs requires the synergistic interplay between the m7GpppN cap structure and the poly(A) tail during initiation. We have developed and characterized a cell-free system from human HeLa cells that recapitulates this important feature, displaying more than one order of magnitude of translational synergism between the cap structure and the poly(A) tail. The stimulation of cap-dependent translation by the poly(A) tail is length-dependent, but not mediated by changes in mRNA stability. Using this system, we investigated the effect of the poly(A) tail on the translation of picornaviral RNAs, which are naturally polyadenylated but initiate translation via internal ribosome entry sites (IRESs). We show that translation driven by the IRESs of poliovirus (PV), encephalomyocarditis virus (EMCV), and hepatitis A virus is also significantly augmented by a poly(A) tail, ranging from an approximately 3-fold stimulation for the EMCV-IRES to a more than 10-fold effect for the PV IRES. These results raise interesting questions concerning the underlying molecular mechanism(s). The cell-free system described here should prove useful in studying these questions as well as providing a general biochemical tool to examine the translation initiation pathway in a more physiological setting.
Assuntos
Regiões 5' não Traduzidas/fisiologia , Sistema Livre de Células , Regulação Viral da Expressão Gênica , Iniciação Traducional da Cadeia Peptídica , Picornaviridae/genética , Poli A/metabolismo , RNA Mensageiro/genética , RNA Viral/genética , Vírus da Encefalomiocardite/genética , Células HeLa , Hepatovirus/genética , Humanos , Poliovirus/genética , Poli A/química , Capuzes de RNA , RNA Mensageiro/química , RNA Viral/química , Ribossomos/fisiologiaRESUMO
Human cytomegalovirus UL25 codes for a structural phosphoprotein of 85 kDa (C. J. Baldick and T. Shenk, J. Virol. 70:6097-6105, 1996; M. C. Battista et al., J. Virol. 73:3800-3809, 1999). In this study we analyzed the intracellular and intraviral localization of pUL25 by confocal and immunoelectron microscopy and found that pUL25 is a component of the viral tegument and the dense body matrix, acquired during the late cytoplasmic phase of virus maturation.
Assuntos
Citomegalovirus/química , Proteínas Virais/análise , Proteínas Estruturais Virais/análise , Linhagem Celular , Citomegalovirus/ultraestrutura , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Fosfoproteínas/análise , Fosfoproteínas/biossíntese , Proteínas Virais/biossíntese , Proteínas Estruturais Virais/biossínteseRESUMO
Human cytomegalovirus (HCMV) UL25 has recently been found to encode a new structural protein that is present in both virion and defective viral particles (C. J. Baldick and T. Shenk, J. Virol. 70:6097-6105, 1996). In the present work a polyclonal antibody was raised against a prokaryotic pUL25 fusion protein in order to investigate the biosynthesis and localization of the UL25 product (pUL25) during HCMV replication in human fibroblasts. Furthermore, pUL25 was transiently expressed in its native form and fused to the FLAG epitope, in COS7 and U373MG cells, in order to compare the properties of the isolated protein and that produced during infection. Immunoblotting analysis revealed a group of polypeptides, ranging from 80 to 100 kDa, in both transfected and infected cells; in vivo labeling experiments with infected cells demonstrated they are posttranslationally modified by phosphorylation. The transcriptional analysis of the UL25 open reading frame combined with the study of pUL25 biosynthesis showed true late kinetics for this protein in infected human fibroblasts. By indirect immunofluorescence both recombinant and viral pUL25 were detected exclusively in the cytoplasm of transfected or infected cells. Interestingly, pUL25 was shown to localize in typical condensed structures in the perinuclear region as already observed for other HCMV tegument proteins. Colocalization of ppUL99 in the same vacuoles suggests that these structure are endosomal cisternae, which are proposed to be a preferential site of viral particle envelopment. Our data suggest that pUL25 is most likely a novel tegument protein and possibly plays a key role in the process of envelopment.
Assuntos
Citomegalovirus/metabolismo , Proteínas Virais/metabolismo , Animais , Células COS , Citomegalovirus/genética , Epitopos , Expressão Gênica , Humanos , Camundongos , Oligopeptídeos , Peptídeos , Células Procarióticas , Processamento de Proteína Pós-Traducional , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Frações Subcelulares , Fatores de Tempo , Transcrição Gênica , Células Tumorais Cultivadas , Proteínas Virais/genéticaRESUMO
We developed a new cytomegalovirus (CMV) immunoglobulin M (IgM) immunoblot to detect CMV-specific IgM in human sera. The new test contains four viral proteins (vp150, vp82, vp65, and vp28) purified from viral particles and four recombinant proteins (rp150, rp130, rp52, and rp38) purified from Escherichia coli. These antigens were individually loaded onto nitrocellulose strips, and the strips were then used to detect CMV-specific IgM by using a mu-specific conjugate. The new assay was evaluated in parallel with one or two IgM enzyme-linked immunosorbent assays (ELISAs) to test 592 serum samples from different groups of latently or acutely infected individuals. The sensitivity of the new assay with respect to the consensus of two ELISAs was 100%, the specificity was 98.6%, the positive predictive value was 96.9%, and the negative predictive value was 100%. We also evaluated the new test by testing sera from pregnant women and transplant recipients with a known clinical history. Our results suggest that the new test combines high sensitivity with high specificity, characteristics that are mutually exclusive with the other commercially available tests. Furthermore, a statistically significant correlation was observed between the number of IgM-reactive bands and the elevated risk of transmission from CMV-infected pregnant women to their offspring.
Assuntos
Anticorpos Antivirais/sangue , Western Blotting/métodos , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/imunologia , Citomegalovirus/imunologia , Imunoglobulina M/sangue , Adulto , Algoritmos , Especificidade de Anticorpos , Antígenos Virais , Infecções por Citomegalovirus/complicações , Estudos de Avaliação como Assunto , Feminino , Humanos , Troca Materno-Fetal , Transplante de Órgãos , Gravidez , Complicações Infecciosas na Gravidez/diagnóstico , Complicações Infecciosas na Gravidez/imunologia , Proteínas Recombinantes/imunologia , Proteínas Virais/imunologiaRESUMO
beta2.7 is the major early transcript produced during human cytomegalovirus infection. This abundantly expressed RNA is polysome associated, but no protein product has ever been detected. In this study, a stable peptide of 24 kDa was produced in vitro from the major open reading frame (ORF), TRL4. Following transient transfection, the intracellular localization was nucleolar and the expression was posttranscriptionally inhibited by the 5' sequence of the transcript, which harbors two short upstream ORFs.
Assuntos
Citomegalovirus/genética , Processamento Pós-Transcricional do RNA , RNA Mensageiro/genética , Proteínas Virais/genética , Animais , Northern Blotting , Células COS , Linhagem Celular , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Fases de Leitura Aberta , Biossíntese de Proteínas/genética , RNA Viral/genéticaRESUMO
Structural and phenotypic modifications of rat thymocytes from birth up to one year of age, i.e. during maturation and at the beginning of the involutive process of the thymus are described. Since the biological significance and the mechanisms of thymic involution are still a matter of debate, this study aims at clarifying the complexity of the compensatory events occurring during this relatively neglected period of time. Thymuses from Sprague-Dawley rats were analyzed morphologically and morphometrically by light and electron microscopy. At the same time, thymocyte subsets, isolated from the same animals, were characterized by flow cytometry according to physical parameters and phenotypic markers. Results indicate that major changes occur during the first month from birth and from six months onward. In particular, already during the first weeks after birth, thymocytes undergo a slight reduction of mitoses associated with an increased number of apoptoses. Moreover, during the same period of time, flow cytometry revealed an expansion of small thymocytes and changes in thymocyte subsets such as increase of CD4+CD8+ and CD5+alpha(beta)TCR- and a decrease of CD4-CD8-, CD4-CD8+ cells. The thymus of adult rats was characterized by time-dependent decrease of both mitoses and apoptoses, progressive physical disconnection among cells, increase of necrotic areas and fibrosis. Around one year of age tissue changes were associated with a dramatic reduction of the population of large thymocytes and the rise of numerous small thymocytes that were unexpectedly negative for all tested markers. By contrast, medium-size thymocytes exhibited a marked decrease of CD4+CD8+ and CD5+alpha(beta)TCR- subsets. In conclusion, our data indicate that thymus undergoes, with time, a complex remodeling and suggest that thymic involution is not only a simple shrinkage of the organ but rather the result of a series of compensatory mechanisms among different cell populations in a setting of progressive involution.
Assuntos
Subpopulações de Linfócitos T/citologia , Timo/citologia , Timo/crescimento & desenvolvimento , Envelhecimento , Animais , Antígenos de Diferenciação de Linfócitos T/análise , Apoptose , Adesão Celular , Diferenciação Celular , Tamanho Celular , Fibroblastos , Citometria de Fluxo , Macrófagos , Masculino , Microscopia Eletrônica , Mitose , Ratos , Ratos Sprague-Dawley , Subpopulações de Linfócitos T/metabolismo , Timo/ultraestruturaRESUMO
OBJECTIVE: Human cytomegalovirus (HCMV) is often isolated from HIV-1-infected patients and the two viruses can infect the same cell type giving rise to direct bidirectional interactions. Whereas the long terminal repeat (LTR) transactivation ability of HCMV immediate early gene (IE1/IE2) is well documented, no information is available on the possible role of other HCMV proteins. In this study, the activity of ppUL44, an early DNA-binding protein, on HIV LTR transactivation was investigated. METHODS: HIV LTR transactivation by ppUL44 in presence or absence of HIV-1 Tat and HCMV IE1/IE2 was determined in J-Jhan and U973 cells through transient transfection experiments with a series of different expression vectors. Some experiments were also performed on U373-MG astrocytoma cells permanently transfected with UL44 or with another HCMV gene used as a control (UL55). RESULTS: The basal transactivation activity of the HIV LTR was not influenced by the presence of ppUL44. On the contrary, the transactivation observed in the presence of Tat, IE1/IE2 or both factors in synergy was strongly downregulated by ppUL44 in a dose-dependent manner. Deletion constructs of ppUL44 demonstrated that the region of the molecule responsible for the inhibition of the LTR is located within the last 114 amino acids at the carboxyl-terminal region. CONCLUSIONS: The results obtained indicate that within the last 114 amino acids of ppUL44 there is a domain that has a negative effect on the ability of HIV-1 LTR to be activated by both its autologous transactivator Tat and the heterologous transactivator HCMV IE1/IE2 functioning individually or synergistically.
Assuntos
Citomegalovirus/genética , Proteínas de Ligação a DNA/genética , Repetição Terminal Longa de HIV/genética , HIV-1/genética , Glicoproteínas de Membrana , Ativação Transcricional , Proteínas do Envelope Viral , Proteínas Virais/genética , Astrocitoma , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/fisiologia , Regulação para Baixo , Imunofluorescência , Genes tat , Vetores Genéticos , Humanos , Proteínas Imediatamente Precoces/genética , Luciferases/análise , Fases de Leitura Aberta , Plasmídeos , Deleção de Sequência , Transativadores/genética , Transfecção , Células Tumorais Cultivadas , Proteínas Virais/química , Proteínas Virais/fisiologiaRESUMO
The present investigation has been performed to better characterize, in vitro, normal aponeurotic cells in comparison with dermal fibroblasts and with cells derived from Dupuytren's affected aponeuroses. Cells were cultured in monolayer and/or into three-dimensional collagen gels. Cell structure, adhesion, and spreading capability on different substrates, as well as integrin expression were investigated by light and electron microscopy and by flow cytometry. Cell-matrix interactions were also analyzed by gel retraction experiments in the presence, or absence, of RGD peptides and anti-integrin antibodies. Normal aponeurotic cells, compared with dermal fibroblasts, exhibited in vitro peculiar structural features, which were substantially maintained in Dupuytren's aponeurotic cells, irrespective of the substrate they were grown on. By contrast, the aponeurotic cell behavior was different in normal and diseased cells, these latter approaching that of dermal fibroblasts. Normal aponeurotic cells, in fact, were characterized by low efficiency in retracting the collagen gel, low alpha 2, alpha 1, and alpha 5 integrin subunit expression and low adhesion properties onto collagen and fibronectin, whereas cells isolated from the aponeuroses of Dupuytren's patients exhibited higher capability of retracting the collagen gel, increased adhesion properties toward collagen and fibronectin, and higher levels of integrin expression. No differences were observed between dermal fibroblasts from Dupuytren's patients or from normal subjects. These in vitro results are consistent with those previously obtained in situ, suggesting that palmar aponeurotic cells have a peculiar phenotype and that changes in cell-matrix interactions occur in Dupuytren's contracture. Moreover, by comparing data obtained from the retracted fibrotic cords and the still clinically unaffected aponeuroses of the same patients, it may be noted that Dupuytren's disease is not only confined to the clinically involved branches, but includes the whole aponeurosis of the affected hand.
Assuntos
Contratura de Dupuytren/fisiopatologia , Integrinas/fisiologia , Pele/citologia , Tendões/citologia , Tendões/fisiologia , Ferimentos e Lesões/fisiopatologia , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Colágeno , Contratura de Dupuytren/patologia , Matriz Extracelular/patologia , Matriz Extracelular/fisiologia , Matriz Extracelular/ultraestrutura , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/fisiologia , Citometria de Fluxo , Mãos , Humanos , Integrinas/análise , Cinética , Oligopeptídeos/farmacologia , Pele/efeitos dos fármacos , Tendões/efeitos dos fármacos , Ferimentos e Lesões/patologiaRESUMO
Surgical techniques of sub-total reconstructive laryngectomies can often prevent the serious impairment of total laryngectomy without having to relinquish oncological radicality. The aim of the present work has been to report on the experience in this field accrued in the ENT Department of the University of Modena from 1987 to 1992. During this period 54 subtotal laryngectomies were performed. Of these, 13 were crico-hyoido-epiglotto-pexies (C.H.E.P.) and the remaining 41 were crico-hyoido-pexies (C.H.P.). The criteria suggested in the literature was adopted for tumor evaluation, surgical indications and contraindications. All the patients had a follow-up of at least 2 years and 31 of them have had at least 5 years of follow-up. There were 9 deaths: 3 due to intervening illnesses, 2 from second primary tumors and 4 from tumor and/or node recurrences. The overall survival was 83.3% at 2 years and 77.6% at 5 years. Determinate survival (ruling out those who had died because of intervening illnesses) were 88.2% and 80%, respectively. There were 11 neoplastic repetitions of which 2 were of the primary tumor, 2 of the primary tumor plus cervical metastases, and 7 of cervical metastases alone. Recovery surgery was performed in 9 patients, 5 of whom are still alive and disease free. Functional recovery (respiration, deglutition) took place slightly earlier in C.H.E.P. than in C.H.P. but in both cases this could be shortened, particularly by introducing a rehabilitative protocol during the immediate post-operative period. In no case did it prove necessary to perform a total laryngectomy to avoid "ab ingestis" problems and only one patient has a permanent tracheostomy.
Assuntos
Neoplasias Laríngeas/cirurgia , Laringectomia , Seguimentos , Humanos , Neoplasias Laríngeas/mortalidade , Neoplasias Laríngeas/patologia , Estudos Retrospectivos , Taxa de SobrevidaRESUMO
We have previously shown that single or multiple epitopes of the major human cytomegalovirus (HCMV) antigens, produced as fusion proteins in prokaryotes can be valuable diagnostic material in the serology of HCMV infection. In this work we moved to a eukaryotic system, to produce one of the most immunogenic HCMV antigens, ppUL44 (also called pp52 due to its apparent molecular size on acrylamide gels), as a non-fusion protein, in an attempt to eliminate some non-specific reactivity of human sera with bacterial carrier proteins. We expressed the DNA encoding ppUL44 in a highly efficient expression system based on the methylotrophic yeast, Pichia pastoris. Good levels of intracellular, soluble pp52 were produced. We observed an indistinguishable pattern of the yeast pp52 from the viral native protein in immunoblotting and a good reactivity with human sera.
Assuntos
Antígenos Virais/genética , Citomegalovirus/genética , Proteínas de Ligação a DNA/genética , Expressão Gênica , Vetores Genéticos , Pichia/genética , Proteínas Virais/genética , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Antígenos Virais/imunologia , Clonagem Molecular , Citomegalovirus/imunologia , Proteínas de Ligação a DNA/imunologia , Humanos , Proteínas Mitocondriais , Oxirredutases/genética , Proteínas de Plantas , Plasmídeos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Transformação Genética , Proteínas Virais/imunologiaRESUMO
The avascular paraganglioma described in this article appears to be the second such tumor reported in the international literature and the first to be reported in the tympanojugular region. Despite a highly suggestive history and clinical appearance, the tumor showed no signs of vascularization on radiologic studies. The pathologic postoperative study confirmed the diagnosis of paraganglioma with extensive stromal fibrosclerosis and without the typical well-vascularized thin fibrous septa. In the authors' opinion, this observation is notable because of the difficulties encountered in the correct diagnostic interpretation of an avascular mass in the tympanojugular region. In such cases, the possibility of a paraganglioma should always be considered.
Assuntos
Neoplasias da Orelha/diagnóstico , Orelha Média , Tumor do Glomo Jugular/diagnóstico , Angiografia , Capilares/patologia , Diagnóstico Diferencial , Neoplasias da Orelha/irrigação sanguínea , Neoplasias da Orelha/patologia , Neoplasias da Orelha/cirurgia , Orelha Média/patologia , Orelha Média/cirurgia , Feminino , Tumor do Glomo Jugular/irrigação sanguínea , Tumor do Glomo Jugular/patologia , Tumor do Glomo Jugular/cirurgia , Perda Auditiva Condutiva/diagnóstico , Perda Auditiva Condutiva/patologia , Perda Auditiva Condutiva/cirurgia , Humanos , Imageamento por Ressonância Magnética , Pessoa de Meia-IdadeRESUMO
In order to give detailed structural and quantitative evaluations for some of the most important dermal constituents such as collagen, elastic fibres and mesenchymal cells, and for the non-structured extracellular matrix, we performed ultrastructural investigations on dermal biopsies from 50 healthy caucasian subjects aged from 6 fetal months to 83 years. Striking changes were observed, mainly in the perinatal period, for collagen, elastin and mesenchymal cells and, after 50 years of age, for collagen and elastin. Only slight or negligible differences were noted between males and females and in skin specimens taken from different parts of the body but similarly exposed to environmental factors (i.e. UV radiation). Modifications of the non-structured extracellular matrix appeared to be the consequence of changes affecting the other components. The results, therefore, emphasize the importance of the ageing factor in connective tissue metabolism and give further information on both qualitative and quantitative characteristics of normal human dermis.