RESUMO
The B cell antigen receptor (BCR) serves both to initiate signal transduction cascades and to target antigen for processing and presentation by MHC class II molecules. How these two BCR functions are coordinated is not known. Recently, sphingolipid- and cholesterol-rich plasma membrane lipid microdomains, termed lipid rafts, have been identified and proposed to function as platforms for both receptor signaling and membrane trafficking. Here we show that upon cross-linking, the BCR rapidly translocates into ganglioside G(M1)-enriched lipid rafts that contain the Src family kinase Lyn and exclude the phosphatase CD45R. Both Igalpha and Lyn in the lipid rafts become phosphorylated, and subsequently the BCR and a portion of G(M1) are targeted to the class II peptide loading compartment. Entry into lipid rafts, however, is not sufficient for targeting to the antigen processing compartments, as a mutant surface Ig containing a deletion of the cytoplasmic domain is constitutively present in rafts but when cross-linked does not internalize to the antigen processing compartment. Taken together, these results provide evidence for a role for lipid rafts in the initial steps of BCR signaling and antigen targeting.
Assuntos
Antígenos Comuns de Leucócito/imunologia , Lipídeos de Membrana/imunologia , Receptores de Antígenos de Linfócitos B/imunologia , Receptores de Antígenos de Linfócitos T/imunologia , Transdução de Sinais/imunologia , Animais , Colesterol/imunologia , Reagentes de Ligações Cruzadas , Gangliosídeo G(M1)/metabolismo , Antígenos de Histocompatibilidade Classe II/fisiologia , Peroxidase do Rábano Silvestre/farmacocinética , Linfoma de Células B , Camundongos , Mutagênese , Receptores de Antígenos de Linfócitos B/química , Proteínas Recombinantes/imunologia , Deleção de Sequência , Esfingolipídeos/imunologia , Células Tumorais Cultivadas , Domínios de Homologia de srcRESUMO
The multichain immune recognition receptors (MIRRs), including the T cell and B cell antigen receptors and the high affinity receptor for IgE, play an important role in immune cell signaling. The MIRRs have no inherent kinase activity, but rather associate with members of the Src-family kinases to initiate signaling. Although a great deal is understood about the biochemical cascades triggered by MIRRs, the mechanism by which signaling is initiated was not known. The evidence now indicates that the Src-family kinases are concentrated in cholesterol- and sphingolipid-rich membrane microdomains, termed lipid rafts, that exclude the MIRRs. Upon ligand-induced crosslinking the MIRRs translocate into rafts where they are phosphorylated. The MIRRs subsequently form highly ordered, polarized structures termed immunological synapses that provide for prolonged signaling. An understanding of the biochemical composition of rafts and synapses and the mechanisms by which these form should lend insight into the regulation of immune cell activation.
Assuntos
Microdomínios da Membrana/fisiologia , Receptores Imunológicos/fisiologia , Transdução de Sinais/fisiologia , Motivos de Aminoácidos , Animais , Antígenos CD/imunologia , Antígenos CD19/imunologia , Linfócitos B/química , Linfócitos B/imunologia , Toxinas Bacterianas/metabolismo , Antígenos CD28/imunologia , Antígeno CD48 , Diferenciação Celular , Colesterol/análise , Glicoesfingolipídeos/análise , Herpesvirus Humano 4/fisiologia , Humanos , Capeamento Imunológico , Ligantes , Ativação Linfocitária , Substâncias Macromoleculares , Lipídeos de Membrana/análise , Microdomínios da Membrana/química , Microdomínios da Membrana/enzimologia , Fosforilação , Proteína Quinase C/fisiologia , Processamento de Proteína Pós-Traducional , Estrutura Terciária de Proteína , Transporte Proteico , Agregação de Receptores , Receptores de Complemento 3d/imunologia , Receptores Imunológicos/química , Esfingolipídeos/análise , Linfócitos T/química , Linfócitos T/imunologia , Proteínas da Matriz Viral/fisiologia , Quinases da Família src/fisiologiaRESUMO
Brain abscess caused by Pseudallescheria boydii is a highly lethal infection, usually seen in immunosuppressed patients. Five patients with P. boydii brain abscesses are described. Four of these patients acquired their infection after near-drowning; 1 patient developed an abscess after penetrating head trauma. Two patients survived their infections, which included involvement of other body sites (lung, eye, bone) as well as multiple undrained brain abscesses, after prolonged courses of high-dose parenteral miconazole (80-90 mg/kg/d). Progressive increases in miconazole dosage during the treatment periods were required to produce serum levels above the minimum inhibitory concentrations of the fungal isolates.
Assuntos
Abscesso Encefálico/etiologia , Afogamento , Miconazol/uso terapêutico , Micetoma/etiologia , Ressuscitação , Adulto , Abscesso Encefálico/diagnóstico por imagem , Abscesso Encefálico/tratamento farmacológico , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Micetoma/diagnóstico por imagem , Micetoma/tratamento farmacológico , Órbita/lesões , Tomografia Computadorizada por Raios X , Ferimentos Penetrantes/complicaçõesRESUMO
For mature B cells, the encounter with foreign antigen results in the selective expansion of the cells and their differentiation into antibody secreting cells or memory B cells. The response of mature B cells to antigen requires not only antigen binding to and signaling through the B cell antigen receptor (BCR) but also the processing and presentation of the BCR bound antigen to helper T cells. Thus, in mature B cells, the ability to process and present antigen to helper T cells plays a critical role in determining the outcome of antigen encounter. In immature B cells, the binding of antigen results in negative selection of the B cell, inducing apoptosis, anergy or receptor editing. Negative selection of immature B cells requires antigen induced signaling through the BCR, analogous to the signaling function of the BCR in mature B cells. However, the role of class II antigen processing and presentation in immature B cells is less well understood. Current evidence indicates that the ability to process and present antigen bound to the BCR is a late acquisition of developing B cells, suggesting that during negative selection B cells may not present BCR bound antigen and interact with helper T cells. However, the expression of class II molecules is an early acquisition of B cells and recent evidence indicates that the expression of class II molecules early in development is required for the generation of long lived mature B cells. Here we review our current understanding of the processing and presentation of antigen by mature B cells and the role for antigen processing and class II expression during B cell development.
Assuntos
Apresentação de Antígeno/imunologia , Linfócitos B/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Animais , Linfócitos B/citologia , Diferenciação Celular , HumanosRESUMO
Epstein-Barr Virus (EBV) infects B-lymphocytes circulating through the oral epithelium and establishes a lifelong latent infection in a subset of mature-memory B cells. In these latently infected B cells, EBV exhibits limited gene expression with the latent membrane protein 2A (LMP2A) being the most consistently detected transcript. This persistent expression, coupled with many studies ofthe function of LMP2A in vitro and invivo, indicates that LMP2A is functioning to control some aspect of viral latency. Establishment and maintenance of viral latency requires exquisite manipulation of normal B cell signaling and function. LMP2A is capable of blocking normal B cell signal transduction in vitro, suggesting that LMP2A may act to regulate lytic activation from latency in vivo. Furthermore, LMP2A is capable of providing B cells with survival signals in the absence of normal BCR signaling. These data show that LMP2A may help EBV-infected cells to persist in vivo. This review discusses the advances that have been made in our understanding of LMP2A and the effects it has on B cell development, activation, and viral latency.
Assuntos
Linfócitos B/imunologia , Proteínas da Matriz Viral/imunologia , Animais , Transformação Celular Viral , Infecções por Vírus Epstein-Barr/imunologia , Infecções por Vírus Epstein-Barr/virologia , Expressão Gênica , Genes Virais , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/imunologia , Herpesvirus Humano 4/patogenicidade , Humanos , Camundongos , Camundongos Transgênicos , Modelos Imunológicos , Mutação , Transdução de Sinais , Proteínas da Matriz Viral/química , Proteínas da Matriz Viral/genéticaRESUMO
Clinical responses for anticancer agents are based upon tumor regression. We have investigated the potential of glycineamide ribonucleotide transformylase (GAR TFase) inhibitors to produce regressions in multiple preclinical models of colon carcinoma. The growth of multicellular tumor spheroids of WiDr human colon carcinoma was inhibited by the GAR TFase inhibitors 5-deazaacyclotetrahydrofolate (5-DACTHF), its 2'-fluoro, 3'-fluoro, 10-deaza, and 10-thia analogs as well as 5,10-dideazatetrahydrofolate, but none of the compounds caused spheroid regressions. By contrast, complete spheroid disruption was observed with exposure to etoposide, m-AMSA (amsacrine), piritrexim, or 2-desamino-2-methyl-10-propargyl-5,8-dideazafolate (DMPDDF). Light microscopy of the spheroids treated with either 5-DACTHF or DMPDDF suggested that the reason for the difference is extensive cell kill throughout the spheroid in the presence of DMPDDF compared with little or no kill, over that found in controls, with 5-DACTHF. Treatment of spheroids with 5-DACTHF in the presence of 1 microM hypoxanthine resulted in no significant reversal of growth inhibition; 50% reversal required 10 microM hypoxanthine. The spheroid studies were extended to in vivo studies examining the effects of 5-DACTHF on established WiDr and colon 38 tumors. The results showed that, in contrast to melphalan, which produced cures and tumor regressions, 5-DACTHF produced reversible growth inhibition with no significant regression of tumors. The results predict that clinical response, typically measured by tumor regression, may be rare following single agent therapy with inhibitors of de novo purine biosynthesis.
Assuntos
Neoplasias do Colo/patologia , Hidroximetil e Formil Transferases , Purinas/metabolismo , Aciltransferases/antagonistas & inibidores , Adenocarcinoma/patologia , Animais , Divisão Celular/efeitos dos fármacos , Ácido Fólico/análogos & derivados , Ácido Fólico/farmacologia , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Fosforribosilglicinamido Formiltransferase , Tetra-Hidrofolatos/farmacologia , Células Tumorais CultivadasRESUMO
OBJECTIVE: To study the effect of a portable HEPA-filtered air exhaust system (Stackhouse Freedom Surgical Helmet System) on airborne microbial contamination in a modern conventional operating room. DESIGN AND SETTING: Microbial air sampling was done with a two-stage Anderson sampler at the wound site during 46 total joint replacements. All operations were performed by the same surgeon in the same operating room at a large community hospital. RESULTS: In 18 cases done without air exhaust hoods, the number of bacterial and fungal colony-forming units (CFU) ranged from 0.6 to 11.7 (mean, 3.6). Air sampling during 28 operations with the operating team in air exhaust hoods revealed a mean of 3.6 CFU (range, 0 to 11.4). Bacterial CFU averaged 3.4 without hoods and 3.2 with exhaust hoods. Coagulase-negative staphylococci were the most common isolates (48% of isolates with hood, 55% without hood). No infections occurred. CONCLUSION: We concluded that these air exhaust hoods did not lower airborne microbial contamination detectable with this air sampling method, as compared to standard head cover and mask, in a modern conventional operating room.
Assuntos
Microbiologia do Ar , Salas Cirúrgicas , Ventilação/instrumentação , Contagem de Colônia Microbiana , Monitoramento Ambiental , Estudos de Avaliação como Assunto , Humanos , Prótese Articular , Ventilação/normasRESUMO
A patient was referred to our hospital with a tentative diagnosis of fungal endocarditis based upon clinical symptoms, suggestive travel history, and microscopic visualization in blood cultures of gram-negative bulbous filaments that appeared to be fungal elements. Subcultures of the blood culture bottles were unsuccessful on all media with the exception of blood agar plates, which had been cross-streaked with Staphylococcus aureus. These plates grew vitamin B6-dependent streptococci. This nutritionally variant organism was determined by biochemical tests to be Streptococcus mitis (mitior). It had a penicillin MIC and MBC of 0.015 micrograms/mL and 0.03 micrograms/mL, respectively and streptomycin MIC and MBC of 0.78 micrograms/mL and 1.56 micrograms/mL, respectively. The patient was treated with these two agents and recovered. We stress the importance of suspecting vitamin B6-dependent streptococci, even when gram stains may suggest presence of other microorganisms.
Assuntos
Endocardite Bacteriana/microbiologia , Micoses/diagnóstico , Infecções Estreptocócicas/diagnóstico , Diagnóstico Diferencial , Humanos , Masculino , Pessoa de Meia-Idade , Piridoxina/metabolismo , Streptococcus/metabolismoRESUMO
Notexin, a myotoxic phospholipase, was used to induce focal necrosis in the sartorius muscles of normal mixed-breed adult dogs and in 12-week-old beagles. Notexin injury caused pathologic changes similar to those of Duchenne muscular dystrophy (DMD) and its canine homologue, golden retriever muscular dystrophy (GRMD). All three conditions are characterized by increased serum creatine kinase (CK) levels, sarcolemmal defects, delta lesions, hyaline degeneration of myofibers, calcium-positive myofibers, and minimal effects on neurovascular structures. Four and 24 h after exposure to notexin, serum CK levels were elevated, and many myofibers were necrotic. In addition, by 24 h the necrotic areas were heavily invaded by mononuclear cells, and calcium-positive myofibers were prominent. Capillaries appeared intact even in areas of marked myonecrosis. Massive cellular infiltrate and myotube formation was evident at 3 days post injury. By 7 days, most affected fascicles were occupied by small immature myofibers. Regeneration was largely complete at 21 days. Our results suggest that notexin-induced muscle injury in dogs will be useful in the evaluation of potential therapies for DMD such as myoblast transplantation.
Assuntos
Venenos Elapídicos/toxicidade , Músculos/patologia , Neurotoxinas/toxicidade , Animais , Capilares/efeitos dos fármacos , Cães , Relação Dose-Resposta a Droga , Microscopia Eletrônica , Músculos/efeitos dos fármacos , Músculos/ultraestrutura , NecroseRESUMO
The interaction between Mycoplasma gallisepticum (MG) and the tracheal mucosa of the young chicken was studied. The use of a selective plating method permitted differentiation between a pathogenic tylosin-resistant strain (227) and a less pathogenic tylosin-sensitive vaccine strain (F). Both MG strains adhered to the tracheal mucosa and colonized equally well. In mixed infection, the presence or absence of the second strain did not change the efficiency of colonization by either strain. When chickens were exposed to the vaccine strain 24 hr or 2 weeks before superinfection by the pathogen, there was no significant reduction in the efficiency of superinfection, despite the presence of 10(6) colony-forming units of MG strain F in the trachea. However, chickens had an increased ability to resist superinfection 5 weeks after exposure via the air sac. These results suggest that the biological mechanism underlying protection of F-strain-vaccinated chickens against adventitious infection by the homologous species does not involve competition for adherence sites or blockage by prior colonization.
Assuntos
Galinhas/microbiologia , Infecções por Mycoplasma/veterinária , Doenças das Aves Domésticas/microbiologia , Doenças da Traqueia/veterinária , Animais , Resistência Microbiana a Medicamentos , Leucomicinas/farmacologia , Mycoplasma/efeitos dos fármacos , Mycoplasma/patogenicidade , Infecções por Mycoplasma/patologia , Doenças das Aves Domésticas/patologia , Doenças da Traqueia/microbiologia , Doenças da Traqueia/patologiaRESUMO
Fledgling budgerigars from aviaries in Georgia and Texas were reported to have high rates of mortality. Affected birds died acutely and exhibited abdominal distention and reddening of the skin. Postmortem lesions were hydropericardium, enlarged heart and liver with areas of necrosis, and swollen, congested kidneys. Histologic examination of a variety of tissues revealed cells with enlarged nuclei containing inclusions. Electron micrographs revealed the presence of viral particles 42 to 49 nm in diameter in the nuclei of epithelial cells of the renal tubule.
Assuntos
Doenças das Aves/patologia , Periquitos , Psittaciformes , Viroses/veterinária , Animais , Rim/patologia , Fígado/patologia , Viroses/patologiaRESUMO
A virus suspected of causing high death rates in fledgling budgerigars in Georgia and Texas aviaries was isolated in budgerigar embryo fibroblasts inoculated with tissue homogenates from affected birds. Virus was most easily recovered from tissues containing many intranuclear inclusion bodies. Cytopathic effect on fibroblasts of all four isolates was characterized by a swollen nucleus followed by rounding and detachment of the affected cell from the monolayer. Properties suggesting the B-931 isolate belongs to the papovaviridae family are (1) presence of DNA; (2) insensitivity to treatment with CHCl3; and (3) presence of cubic viral particles 42 to 49 nm in diameter in the nucleus of infected chicken embryo fibroblasts. The isolate did not hemagglutinate erythrocytes of chickens, turkeys, budgerigars, guinea pigs, or type O humans and was basically stable against heating and freeze-thawing. An examination of fledgling budgerigars from infected aviaries demonstrated that sick birds carried more virus than healthy birds.
Assuntos
Papillomaviridae/isolamento & purificação , Periquitos/microbiologia , Polyomaviridae , Psittaciformes/microbiologia , Animais , Linhagem Celular , Embrião de Galinha , Clorofórmio/farmacologia , Efeito Citopatogênico Viral , Congelamento , Temperatura Alta , Papillomaviridae/crescimento & desenvolvimento , Papillomaviridae/patogenicidade , Virulência , Replicação ViralRESUMO
Zebrafish (Brachydanio rerio) have become an important model system for studying vertebrate embryonic development and gene function through manipulation of genotype and characterization of resultant phenotypes. An established research zebrafish colony without substantial disease problems for more than 7 years of operation began experiencing appreciable mortalities in November of 1997. Young fish (fry), from five to 24 days after hatching, spontaneously developed elongate strands of organic material protruding from the mouth, operculum, and anal pore, leading workers in the laboratory to describe the infected fish as "bearded." Unlike typical freshwater fish fungal infections, the skin surface did not have evidence of fungal colonization. The disease was associated with progressive lethargy, reduced feeding, and subsequent mortality. From 10 to 100% of the fry in a given tank were affected. Initial examination indicated that the biofilm around the head of affected fry consisted of bundles of septate fungal hyphae, large numbers of mixed bacterial populations, and protozoans. Environmental samples of air and water in the laboratory were obtained to ascertain the source of the infective agent and to isolate and identify the fungus. A fungus identified as Lecythophora mutabilis was isolated repeatedly from infected fish and water samples from infected fish tanks, and from the main laboratory water supply tanks, but not from laboratory air. Some biofilm beards on fish were found to consist of relatively pure bacterial populations, and beards on occasional fish examined in the later part of the study consisted of hyphae and spores of the oomycete genus Aphanomyces. Lecythophora mutabilis did not invade tissues; however, elimination of the epizootic correlated with reduction in the number of L. mutabilis conidia in the water following modification of the laboratory water system by use of new filtration and sterilization systems. We conclude that the dense hyphal strands of L. mutabilis composing the predominant biofilm type, along with mixed bacteria and protozoa, contributed to the die-off in young fry by occluding the oral cavity and/or gills, leading to starvation and/or asphyxiation.
Assuntos
Surtos de Doenças/veterinária , Doenças dos Peixes/mortalidade , Pesqueiros , Micoses/veterinária , Infecções Oportunistas/veterinária , Sordariales/isolamento & purificação , Peixe-Zebra/microbiologia , Microbiologia do Ar , Animais , Antifúngicos/farmacologia , Bactérias/isolamento & purificação , Biofilmes , Filtração , Doenças dos Peixes/microbiologia , Pesqueiros/instrumentação , Brânquias/microbiologia , Massachusetts/epidemiologia , Micoses/microbiologia , Micoses/mortalidade , Infecções Oportunistas/microbiologia , Infecções Oportunistas/mortalidade , Sordariales/efeitos dos fármacos , Esterilização , Microbiologia da Água , Abastecimento de ÁguaRESUMO
Despite improved results following carotid endarterectomy, a small but significant incidence of neurological deficit may follow the operation. Emphasis has deviated from the importance of the technical details of the operative procedure to consideration of the role of physiological factors (e.g., shunt, stump pressure, EEG vigilance, type of anesthesia). The surgical principles outlined in this report were used in a consecutive series of 214 patients undergoing carotid endarterectomy. A neurological deficit was noted in two patients (0.9%) in whom thrombosis of the repair site was proven.
Assuntos
Doenças das Artérias Carótidas/cirurgia , Endarterectomia/métodos , Arteriosclerose Intracraniana/cirurgia , Doenças do Sistema Nervoso/prevenção & controle , Complicações Pós-Operatórias/prevenção & controle , Artérias Carótidas/cirurgia , HumanosRESUMO
Twenty-eight weanling pigs inoculated with sporocysts from an isolate of Sarcocystis suicanis from Georgia were examined at intervals ranging from 2 to 90 days postinoculation (DPI). Merogony was first observed histologically within the heart muscle 12 DPI and within 23 of 35 tissues examined 13 DPI. Most infected cells were "floating" in extravascular spaces and were near intact endothelial cells. In some cases, the infected cell clearly was an endothelial cell comprising a portion of the capillary wall. Immature sarcocysts containing metrocytes were observed in striated muscle 27 DPI, and bradyzoites were detected by digestion techniques 52 DPI. Sarcocysts matured between 27 and 80 DPI, after which thickness of the cyst wall and morphology of bradyzoites changed little. Dissolution of sarcocysts was detected as early as 38 DPI and was accompanied by ingress of plasma cells, lymphocytes, and occasionally, eosinophils. Based on information presented herein, feeder pigs reared on pasture may become infected, and infections mature well within the 100-day period usually considered necessary for production of marketable swine.
Assuntos
Sarcocystis/crescimento & desenvolvimento , Sarcocistose/veterinária , Doenças dos Suínos/parasitologia , Animais , Sarcocystis/patogenicidade , Sarcocystis/ultraestrutura , Sarcocistose/parasitologia , Suínos/parasitologiaRESUMO
Lesions in estuarine finfish are associated with a variety of organisms including parasites and bacterial, viral, and fungal infectious agents. In addition, trauma, suboptimal water quality, and other abiotic stress factors may result in the loss of homeostasis. We have observed solitary ulcerative lesions on menhaden sampled from the Chesapeake Bay, Maryland, the Pimlico River, North Carolina, and the St. Johns River, Florida. Histologically, the lesions demonstrated a marked chronic inflammatory infiltrate and granulomas in response to fungal hyphae throughout large areas of exposed necrotic muscle. Gram-negative rod-shaped bacteria were also observed in the lesions, a common finding in ulcers of aquatic organisms. Similar observations in menhaden and other species have been described previously in the literature as ulcerative mycosis, mycotic granulomatosis, red spot disease, and epizootic ulcerative syndrome. Despite the many different known causes of fish lesions, the popular press and the scientific literature have recently emphasized Pfiesteria piscicida and other Pfiesteria-like dinoflagellates (and their bioactive compounds) as the primary causative agent for finfish lesions, particularly mycotic granulomatous ulcers in Atlantic menhaden. While some laboratory data suggest that Pfiesteria may play a role in field-observed lesions, much more cause-and-effect evidence is needed to determine the importance of other risk factors, both alone or and in combination with Pfiesteria. In order to better understand the etiology of lesion initiation and progression in estuarine finfish, accurate assessments of environmental conditions collected on appropriate temporal and spatial scales, and fish morphological indicators consistent with gross and histological pathologic terminology, should be used for reporting fish lesion observations and kills. Further, this outlook will help to avoid bias and may foster a broader perspective for examining the health of estuarine systems in general.
Assuntos
Doenças dos Peixes/etiologia , Animais , Dinoflagellida , Doenças dos Peixes/parasitologia , Doenças dos Peixes/patologia , Peixes , Florida , Granuloma/etiologia , Granuloma/patologia , Granuloma/veterinária , Maryland , North Carolina , Fatores de RiscoRESUMO
Borrelia anserina (Sakharoff) was successfully grown in a liquid medium (Barbour-Stoenner-Kelly) for 39 passages. By the 12th serial passage in medium, infectivity of B anserina for chicks was lost. Electron microscopy did not reveal structural differences between non-infective and infective cultured organisms. Changes in the protein profiles were found by electrophoresis as the organisms were passed in culture.
Assuntos
Infecções por Borrelia/veterinária , Borrelia/patogenicidade , Galinhas , Doenças das Aves Domésticas/microbiologia , Animais , Proteínas de Bactérias/análise , Borrelia/análise , Borrelia/crescimento & desenvolvimento , Borrelia/ultraestrutura , Infecções por Borrelia/microbiologia , Meios de Cultura , Eletroforese em Gel de Poliacrilamida , Microscopia Eletrônica , Inoculações SeriadasRESUMO
Changes in tracheal epithelial surfaces induced by Mycoplasma infection in vivo and in vitro included release of mucous granules followed by exfoliation of ciliated and nonciliated epithelial cells. Light microscopy, scanning electron microscopy, and transmission electron microscopy confirmed that the loss of cilia from individual cells was infrequent. Epithelial cells typically lost their intercellular connections, rounded up, exfoliated, and then lysed--giving rise to a population of cellular organelles, such as mitochondria and cilia intermixed with mucus to form the exudate found within the tracheal lumen. Repair of the epithelial surface was effected by basilar epithelial cells differentiating and filling in the spaces formed by exfoliated cells. These cells were hypertrophied, nonciliated at 14 days after infection in vivo, and covered with microvilli. In sectioned material obtained during the infection, there was increasing epithelial thickness due to cellular infiltration and edema. Tracheal rings in vitro showed similar changes to those seen in vivo, except that exfoliation was more severe and occurred earlier. In addition, there were no cellular infiltration due to the lack of a vascular supply and only a small amount of mucus due to the smaller number of mucous cells available to release into the tracheal lumen.
Assuntos
Galinhas/microbiologia , Mycoplasma/patogenicidade , Traqueia/ultraestrutura , Animais , Cílios/ultraestrutura , Epitélio/microbiologia , Epitélio/ultraestrutura , Microscopia Eletrônica , Microscopia Eletrônica de Varredura , Mycoplasma/crescimento & desenvolvimento , Técnicas de Cultura de Órgãos , Especificidade da Espécie , Traqueia/microbiologia , VirulênciaRESUMO
OBJECTIVE: To determine the effect of alpha-chymotrypsin treatment on breaking strength and ultrastructural morphology of canine ciliary zonules. SAMPLE POPULATION: Eyes from young random-source dogs from an animal shelter. PROCEDURE: Eyes were obtained immediately after euthanasia of dogs. The enzyme alpha-chymotrypsin was applied to the ciliary zonules of 1 eye of each dog; the other eye was treated with saline solution as a control. The breaking strength of ciliary zonules was measured, using a linear actuator and force transducer. The lenses and ciliary bodies were then analyzed by scanning and transmission electron microscopy. RESULTS: alpha-Chymotrypsin reduced the breaking strength of ciliary zonules by a mean +/- SD 44 (+/- 20)%, compared with that for saline-treated control eyes. Increasing the volume of enzyme further decreased the breaking strength of the zonules. Differences in the appearance of the ciliary body by electron microscopy were not apparent between enzyme- and saline-treated specimens. CONCLUSIONS AND CLINICAL RELEVANCE: Application of alpha-chymotrypsin to enucleated canine eyes at a concentration used in people significantly reduces the breaking strength of canine ciliary zonules without any apparent damage to the ciliary body. alpha-Chymotrypsin may be useful in the removal of subluxated canine lenses and in removal of cataractous lenses in young dogs, in which phacoemulsification often results in appreciable post operative capsular opacification.
Assuntos
Quimotripsina/farmacologia , Corpo Ciliar/fisiologia , Corpo Ciliar/ultraestrutura , Animais , Corpo Ciliar/efeitos dos fármacos , Cães , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/fisiologia , Células Epiteliais/ultraestrutura , Microscopia Eletrônica , Microscopia Eletrônica de Varredura , Resistência à TraçãoRESUMO
OBJECTIVE: To determine histologic and immunohistochemical characteristics of the multifocal adherent plaques that commonly develop on the internal surfaces of the anterior and posterior lens capsules in dogs with cataracts. SAMPLE POPULATION: 31 anterior and 4 posterior capsular specimens collected during lens extraction surgery in dogs with cataracts. PROCEDURE: Specimens were evaluated, using light and transmission electron microscopy. Immunohistochemical techniques were used to localize cytokeratin, vimentin, alpha-smooth muscle-specific actin, fibronectin, tenascin, and transforming growth factor-beta (TGF-beta) within plaques. RESULTS: Histologically, plaques comprised elongated spindle-shaped cells that formed a placoid mass. Cells were embedded in an extracellular matrix containing collagen fibrils, often with duplicated or split basement membranes. Immunohistochemically, normal lens epithelial cells and cells within plaques stained for vimentin. Most cells and some areas of the extracellular matrix within plaques stained for TGF-beta and alpha-smooth muscle-specific actin. Fibronectin and tenascin were also detected in the extracellular matrix. CONCLUSIONS AND CLINICAL RELEVANCE: Canine lens capsular plaques are histologically and immunohistochemically similar to posterior capsule opacification and subcapsular cataracts in humans, which suggests that the canine condition, like the human conditions, is associated with fibrous metaplasia of lens epithelial cells. Transforming growth factor-beta may play a role in the genesis of capsular plaques. Because severity of plaques was correlated with stage of cataract development, earlier surgical removal of cataracts may be useful to avoid complications associated with plaque formation.