RESUMO
Climate change is causing extreme heating events and intensifying infectious disease outbreaks. Animals harbour microbial communities, which are vital for their survival and fitness under stressful conditions. Understanding how microbiome structures change in response to infection and warming may be important for forecasting host performance under global change. Here, we evaluated alterations in the microbiomes of several wild Caenorhabditis elegans isolates spanning a range of latitudes, upon warming temperatures and infection by the parasite Leucobacter musarum. Using 16S rRNA sequencing, we found that microbiome diversity decreased, and dispersion increased over time, with the former being more prominent in uninfected adults and the latter aggravated by infection. Infection reduced dominance of specific microbial taxa, and increased microbiome dispersion, indicating destabilizing effects on host microbial communities. Exposing infected hosts to warming did not have an additive destabilizing effect on their microbiomes. Moreover, warming during pre-adult development alleviated the destabilizing effects of infection on host microbiomes. These results revealed an opposing interaction between biotic and abiotic factors on microbiome structure. Lastly, we showed that increased microbiome dispersion might be associated with decreased variability in microbial species interaction strength. Overall, these findings improve our understanding of animal microbiome dynamics amidst concurrent climate change and epidemics. This article is part of the theme issue 'Sculpting the microbiome: how host factors determine and respond to microbial colonization'.
Assuntos
Microbiota , Parasitos , Animais , RNA Ribossômico 16S/genética , Mudança Climática , TemperaturaRESUMO
During immune responses, antigen-presenting cells (APCs) process antigens and present peptide epitopes complexed with human leukocyte antigen (HLA) molecules. CD4 cells recognize these naturally processed and presented epitopes (NPPEs) bound to HLA class II molecules. Epitope identification is important for developing diagnostic and therapeutic tools for immune-mediated diseases and providing insight into their etiology, but current approaches overlook effects of natural processing on epitope selection. We have developed a technique to identify NPPEs using mass spectrometry (MS) after antigen is targeted onto APCs using a lectin-based antigen delivery system (ADS). We applied the technique to identify NPPEs of the intracellular domain of the type 1 diabetes mellitus-associated (type 1 DM-associated) autoantigen insulinoma-associated-2 (IA-2ic), presented by HLA-DR4 (0401). IA-2ic-derived NPPEs eluted from HLA-DR4 constitute 6 sets of peptides nested around distinct core regions. Synthetic peptides based on these regions bind HLA-DR4 and elicit primary T-cell proliferation frequently in HLA-DR4-positive type 1 DM patients, but rarely in non-HLA-DR4 patients, and in none of the HLA-DR4 nondiabetic controls we tested. This flexible, direct approach identifies an HLA allele-specific map of NPPEs for any antigen, presented by any HLA class II molecule. This method should enable a greater understanding of epitope selection and lead to the generation of sensitive and specific reagents for detecting autoreactive T cells.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Artrite Reumatoide/imunologia , Autoantígenos/imunologia , Diabetes Mellitus Tipo 1/imunologia , Epitopos/imunologia , Antígeno HLA-DR4/imunologia , Ilhotas Pancreáticas/imunologia , Proteínas de Membrana/imunologia , Proteínas Tirosina Fosfatases/imunologia , Adolescente , Adulto , Idoso , Sequência de Aminoácidos , Artrite Reumatoide/genética , Autoanticorpos/análise , Linfócitos B , Linhagem Celular , Membrana Celular/imunologia , Criança , Diabetes Mellitus Tipo 1/genética , Epitopos/química , Epitopos/isolamento & purificação , Europa (Continente) , Genótipo , Humanos , Ativação Linfocitária , Proteínas de Membrana/química , Pessoa de Meia-Idade , Dados de Sequência Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/imunologia , Proteína Tirosina Fosfatase não Receptora Tipo 1 , Proteínas Tirosina Fosfatases/química , Proteínas Tirosina Fosfatases Classe 8 Semelhantes a Receptores , Linfócitos T/imunologia , Estados Unidos , População BrancaRESUMO
Reported frequencies of peripheral blood autoantigen-specific cells in autoimmune diseases are typically low, which could be due to true scarcity or to limitations of in vitro assays. In the present study, antigens were targeted to the antigen-presenting cell (APC) to enhance T cell proliferation, using an antigen delivery system (ADS), consisting of biotinylated anti-IgG, streptavidin and biotinylated antigen. This was able to bind B cells and monocytes and was internalized within 24 hours. T cell proliferation to tetanus toxoid was at least doubled using the ADS compared to conventional assay with antigen in simple solution. To evaluate the ADS in an autoimmune disease, we determined T cell responses to the insulin-dependent diabetes mellitus (IDDM)-associated autoantigen IA-2ic in patients with recent-onset IDDM. When IA-2ic was available conventionally in solution, proliferation was poor, but significantly higher in IDDM patients than control subjects. However, the ADS significantly enhanced proliferation by a mean 3-fold for all subjects, while maintaining the significant difference between IDDM patients and healthy controls. Increases in T cell proliferation via the ADS were due to the recruitment of approximately 3 times the number of CD4 + T cells stimulated in conventional assays. B cell depletion abolished enhancement suggesting that the ADS operates through recruitment of B cells as APCs. This flexible modification of the T cell assay offers greatly enhanced sensitivity for determining the frequency of antigen and autoantigen-reactive T cells.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Antígenos/administração & dosagem , Linfócitos B/imunologia , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Adulto , Animais , Apresentação de Antígeno/efeitos dos fármacos , Apresentação de Antígeno/fisiologia , Células Apresentadoras de Antígenos/efeitos dos fármacos , Antígenos/metabolismo , Autoimunidade/efeitos dos fármacos , Autoimunidade/imunologia , Avidina , Biotina , Linfócitos T CD4-Positivos/imunologia , Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 1/metabolismo , Epitopos , Feminino , Humanos , Imunoglobulina G , Leucócitos Mononucleares/imunologia , Masculino , Ovinos , EstreptavidinaRESUMO
This study examined links between impaired nitric oxide production in the sciatic endoneurium, nerve blood flow, and polyol pathway flux, to test the hypothesis that reduced nerve blood flow might be compromised by competition for NADPH between aldose reductase and nitric oxide synthase. Sciatic nerves of streptozotocin-diabetic rats showed reduced laser Doppler flux (by 51% or 63%; both p<0.05)-indicative of reduced nerve blood flow-and reduced motor nerve conduction velocity (17% in two experiments; p<0.05). Acute interruption of nitric oxide production in the sciatic nerves of control rats, via endoneurial injection of N omega-nitro-D-arginine methyl ester (L-NAME), caused a local reduction (of 64%; p<0.001) in nerve Doppler flux. This was reversed by either L-arginine or sodium nitroprusside. The response to L-NAME was greatly reduced in diabetic rats (only 22% reduction; p<0.01), though both L-arginine and SNP caused marked increases in flux. Chronic inhibition of aldose reductase in diabetic rats (with either sorbinil or imirestat at a range of doses) had little effect on resting sciatic nerve Doppler flux, though both inhibitors normalized conduction velocity. Both aldose reductase inhibitors reduced sorbitol pathway intermediates in a dose-related manner. These findings do not support the proposition that aldose reductase inhibitors normalise conduction velocity by mechanisms dependent upon either normalization of endoneurial nitric oxide or nerve blood flow. Instead, a mechanism based upon more direct effects on axon or Schwann cell function is favoured.
Assuntos
Aldeído Redutase/antagonistas & inibidores , Diabetes Mellitus Experimental/fisiopatologia , Óxido Nítrico/biossíntese , Nervo Isquiático/irrigação sanguínea , Animais , Arginina/farmacologia , Inibidores Enzimáticos/farmacologia , Fluxometria por Laser-Doppler , Masculino , Neurônios Motores/fisiologia , NG-Nitroarginina Metil Éster/farmacologia , Condução Nervosa/efeitos dos fármacos , Óxido Nítrico Sintase/antagonistas & inibidores , Nitroprussiato/farmacologia , Ratos , Ratos WistarAssuntos
Músculos Faciais/fisiopatologia , Doenças Musculares/epidemiologia , Distribuição por Idade , California/epidemiologia , Distribuição de Qui-Quadrado , Criança , Pré-Escolar , Transtornos de Deglutição/complicações , Transtornos de Deglutição/epidemiologia , Etnicidade , Feminino , Humanos , Transtornos da Linguagem/complicações , Transtornos da Linguagem/epidemiologia , Masculino , Má Oclusão/complicações , Má Oclusão/epidemiologia , Músculos da Mastigação/fisiopatologia , Doenças Musculares/complicações , Prevalência , Distribuição por Sexo , Distúrbios da Fala/complicações , Distúrbios da Fala/epidemiologia , Estatísticas não Paramétricas , Inquéritos e QuestionáriosRESUMO
Aminoguanidine (AG) treatment can prevent the development of some functional anomalies in experimentally diabetic rats, possibly via the prevention of a diabetes-induced vascular dysfunction. The acute effects of AG on endothelium-dependent relaxation of aortae in the presence of indomethacin and on pressor responses and prostacyclin release in isolated perfused lungs, were therefore investigated using tissues from control and streptozotocin-diabetic rats. Endothelium-dependent relaxations of aortae were reduced by aminoguanidine (control 20%, and diabetic 25%). For lungs, angiotensin II-induced pressor responses were unaffected by AG, whereas the nitric oxide synthase inhibitor L-NAME caused integrated pressor responses to be increased in lungs from control and diabetic rats (2.0 and 1.8 fold respectively). Individually, AG (1 mM) and L-NAME (10 microM) did not affect total cumulative prostacyclin release by control lungs, whereas significant increases for both were observed for diabetic lungs. In summary, these studies firstly provide evidence that AG can increase prostacyclin release from tissues in vitro, with little effect upon endothelium-dependent vasodilatation, and secondly, that the regulation of vasodilator prostanoid release by the pulmonary circulation of the rat may be altered in experimental diabetes.
Assuntos
Diabetes Mellitus Experimental/fisiopatologia , Guanidinas/farmacologia , Pulmão/efeitos dos fármacos , NG-Nitroarginina Metil Éster/farmacologia , Vasodilatação/efeitos dos fármacos , Acetilcolina/farmacologia , Angiotensina II/farmacologia , Animais , Aorta/efeitos dos fármacos , Diabetes Mellitus Experimental/metabolismo , Endotélio Vascular/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Epoprostenol/metabolismo , Pulmão/metabolismo , Masculino , Fenilefrina/farmacologia , Ratos , Ratos WistarRESUMO
Decreased nerve blood flow may be a pathogenetic factor in diabetic neuropathy. Previously it was shown that insulin treatment, commenced at the onset of streptozotocin-diabetes, prevents the development of a nerve blood flow deficit in the diabetic rat. The present study sought to determine the effect of short-term (one month) and acute (one hour) insulin reversal treatment on nerve blood flow deficits in streptozotocin-diabetes. Sciatic nerve blood flow was assessed using laser Doppler flowmetry. Treatment was initiated after one month of diabetes. One month of reversal insulin treatment ameliorated nerve laser Doppler flux (NDF) deficits; in untreated diabetic rats NDF was 51% of that in control animals (P < 0.01), in insulin-treated diabetic rats NDF was 85% of control values (P < 0.01 vs. untreated diabetic, P < 0.05 vs. control). In association with blood flow increases, we found a significant amelioration of motor (P < 0.05 vs. untreated diabetic) and sensory (P < 0.01 vs. untreated diabetic) nerve conduction velocities but not of exaggerated resistance to hypoxic conduction block. Insulin partially reversed hyperglycaemia and sciatic nerve polyol and sugar levels. In a second experiment, in rats with one month of diabetes, acute infusion of insulin led to a 47% (P < 0.001 vs. pre-insulin values) reduction of plasma glucose. This fall in plasma glucose was accompanied by a 38% (P < 0.05 vs. pre-insulin values) increase in NDF. Sensory nerve conduction velocity was marginally increased (6%, P < 0.05 vs. pre-insulin values) after insulin infusion, but motor conduction velocity was not. The data indicate that insulin can partially reverse deficits in nerve blood flow and conduction in diabetic rats.
Assuntos
Diabetes Mellitus Experimental/tratamento farmacológico , Neuropatias Diabéticas/tratamento farmacológico , Insulina/uso terapêutico , Nervos Periféricos/irrigação sanguínea , Animais , Neuropatias Diabéticas/fisiopatologia , Fluxometria por Laser-Doppler , Masculino , Condução Nervosa , Ratos , Ratos WistarRESUMO
A Pseudomonas aeruginosa strain (UCBPP-PA14) is infectious both in an Arabidopsis thaliana leaf infiltration model and in a mouse full-thickness skin burn model. UCBPP-PA14 exhibits ecotype specificity for Arabidopsis, causing a range of symptoms from none to severe in four different ecotypes. In the mouse model, UCBPP-PA14 is as lethal as other well-studied P. aeruginosa strains. Mutations in the UCBPP-PA14 toxA, plcS, and gacA genes resulted in a significant reduction in pathogenicity in both hosts, indicating that these genes encode virulence factors required for the full expression of pathogenicity in both plants and animals.
Assuntos
ADP Ribose Transferases , Arabidopsis/microbiologia , Toxinas Bacterianas , Doenças das Plantas/microbiologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/patogenicidade , Fatores de Virulência , Animais , Proteínas de Bactérias/genética , Sequência de Bases , Queimaduras/complicações , Exotoxinas/genética , Masculino , Camundongos , Dados de Sequência Molecular , Mutação , Fosfolipases/genética , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/crescimento & desenvolvimento , Virulência/genética , Exotoxina A de Pseudomonas aeruginosaRESUMO
1. The effects of the non-selective endothelin (ET) receptor (ETA/ETB) antagonist, bosentan, on sciatic nerve dysfunction in experimental diabetes were investigated. 2. Rats with 5-6 weeks untreated streptozotocin-diabetes exhibited characteristic slowed motor nerve conduction velocity (mean +/- s.d., 36.6 +/- 3.4 m s-1) and nerve laser Doppler flux (197 +/- 64 arbitrary units) compared to age-matched control animals (42.7 +/- 2.4 m s-1 and 398 +/- 77 arbitrary units, respectively). Preventative treatment of diabetic rats with bosentan at 100 mg kg-1 day-1 p.o. attenuated both these deficits (39.7 +/- 3.0 m s-1 and 305 +/- 56 arbitrary units, respectively) without affecting mean arterial pressure. 3. In control and untreated diabetic rats, ET-1, 1 nmol kg-1 i.v., caused an initial hypotension (duration, 30 +/- 13 and 26 +/- 9 s, respectively; change in mean arterial pressure, -27 +/- 13 and -25 +/- 7 mmHg, respectively) followed by prolonged hypertension (change in mean arterial pressure, 52 +/- 18 and 31 +/- 5 mmHg, respectively). Effectiveness of the chronic bosentan treatment was demonstrated by inhibition of the hypotensive response to ET-1 in treated diabetic rats (duration, 5 +/- 2 s; change in mean arterial pressure, -4 +/- 2 mmHg) although the hypertension was unaltered (change in mean arterial pressure, 32 +/- 9 mmHg). 4. Acute i.v. administration of 10 mg kg-1 bosentan caused variable and transient rises in nerve laser Doppler flux in control (78 +/- 63 arbitrary units) and untreated diabetic rats (93 +/- 77 arbitrary units). Acute bosentan blocked the hypotensive response to subsequent ET-1 administration and attenuated the later hypertension (change in mean arterial pressure, 21 +/-9 mmHg in control, 29 +/- 10 mmHg in diabetic).5. Our results indicate that oral treatment of diabetic rats with an ET receptor antagonist can improves ciatic nerve perfusion and conduction, suggesting that the vasoconstrictor action of endogenous ET may contribute to peripheral nerve dysfunction in experimental diabetes.
Assuntos
Diabetes Mellitus Experimental/fisiopatologia , Antagonistas dos Receptores de Endotelina , Endotelinas/farmacologia , Nervo Isquiático/efeitos dos fármacos , Sulfonamidas/farmacologia , Animais , Glicemia/análise , Glicemia/efeitos dos fármacos , Pressão Sanguínea/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Bosentana , Modelos Animais de Doenças , Endotelinas/administração & dosagem , Frequência Cardíaca/efeitos dos fármacos , Injeções Intravenosas , Fluxometria por Laser-Doppler , Masculino , Condução Nervosa/efeitos dos fármacos , Ratos , Ratos Wistar , Nervo Isquiático/irrigação sanguínea , Nervo Isquiático/fisiologia , Estreptozocina/administração & dosagem , Estreptozocina/toxicidade , Sulfonamidas/administração & dosagemRESUMO
This report characterizes nine new cell lines derived from patients with malignant pleural mesothelioma. The lines were initiated between July 1990 and July 1992 from solid tumors (5 lines) or effusions (4 lines) and had proliferated for a period of at least 2 months without senescence. They were characterized by cell size, doubling time, immunohistochemical analyses, electron microscopy, and chromosomal karyotyping. Growth factor/cytokine elaboration was determined using enzyme-linked immunoassays. The established lines were similar in morphology to their parent tumor (ie, epithelial or sarcomatoid). Cell sizes ranged from 59 to 81 microns, and the doubling times varied from 31 to 65 hours. The lines stained with cytokeratin and showed expected negative staining for adenomarkers including B72.3 and carcinoembryonic antigen. All cell lines exhibited aneuploidy, with modal chromosome numbers between 40 and 81 and had multiple chromosomal aberrations. Significant production of granulocyte-monocyte colony-stimulating factor, leukemia inhibitory factor, platelet-derived growth factor, and interleukin-6 was seen. These new cell lines derived from human mesotheliomas can now be used to aid in the design of innovative treatment strategies.
Assuntos
Mesotelioma , Neoplasias Pleurais , Células Tumorais Cultivadas , Adulto , Idoso , Divisão Celular , Meios de Cultura , Citocinas/biossíntese , Feminino , Humanos , Imuno-Histoquímica , Cariotipagem , Masculino , Mesotelioma/química , Mesotelioma/genética , Mesotelioma/metabolismo , Mesotelioma/patologia , Mesotelioma/ultraestrutura , Pessoa de Meia-Idade , Neoplasias Pleurais/química , Neoplasias Pleurais/genética , Neoplasias Pleurais/metabolismo , Neoplasias Pleurais/patologia , Neoplasias Pleurais/ultraestrutura , Células Tumorais Cultivadas/química , Células Tumorais Cultivadas/metabolismo , Células Tumorais Cultivadas/patologia , Células Tumorais Cultivadas/ultraestruturaRESUMO
In Study 1, the effects of treatment of streptozotocin-diabetic rats with the antioxidants, probucol or vitamin E were compared. Untreated diabetic rats showed a reduction of 45% (p < 0.01) in nerve laser Doppler flux, which was used as an index of nerve blood flow. In diabetic rats treated with either probucol or vitamin E nerve Doppler flux was reduced by only 13 or 16%, respectively (p < 0.01 for either compared to untreated diabetic rats). A second study examined the effects of treatment with evening primrose oil either alone or in combination with probucol. Reduced nerve Doppler flux was reproduced in untreated diabetic rats (47%; p < 0.01). In parallel diabetic groups, nerve Doppler flux was reduced by only 14% with evening primrose oil alone and by 8% with evening primrose oil plus probucol (both p < 0.01 vs untreated diabetic rats). Both treatments were also associated with marked attenuation of motor and sensory nerve conduction velocity deficits. Measurements on plasma from rats showed normalisation of triglyceride levels by probucol treatment without an effect on those of cholesterol in Study 1. In Study 2, the converse was true for evening primrose oil treatment, whilst the combined treatment lowered both plasma triglycerides and cholesterol. This work indicates similar effects of antioxidants and evening primrose oil against reduced nerve Doppler flux and conduction velocity in diabetic rats, with dissimilar actions on plasma triglycerides and cholesterol.
Assuntos
Antioxidantes/uso terapêutico , Diabetes Mellitus Experimental/tratamento farmacológico , Ácidos Graxos Essenciais/sangue , Nervo Isquiático/efeitos dos fármacos , Animais , Colesterol/sangue , Diabetes Mellitus Experimental/fisiopatologia , Ácidos Graxos Essenciais/uso terapêutico , Ácidos Linoleicos , Masculino , Condução Nervosa/efeitos dos fármacos , Condução Nervosa/fisiologia , Oenothera biennis , Doenças do Sistema Nervoso Periférico/tratamento farmacológico , Doenças do Sistema Nervoso Periférico/fisiopatologia , Óleos de Plantas , Probucol/uso terapêutico , Ratos , Ratos Wistar , Nervo Isquiático/fisiopatologia , Triglicerídeos/sangue , Vitamina E/uso terapêutico , Ácido gama-LinolênicoRESUMO
PBLs were isolated from 13 patients with metastatic melanoma. Mixed lymphocyte tumor cell cultures (ML TCs) were established (15 times) by using irradiated HLA-matched (one class I locus) allogeneic melanoma tumor cell lines (13 times) or autologous melanoma tumor cell lines (two times) in medium containing 120 IU/ml IL-2 and 100 IU/ml IL-4. PBLs grew to levels that could be assessed for functional reactivity 9 of 15 times. In seven of nine cases, CD3+CD8+ CTLs grew from MLTCs that were tumor specific; five were restricted by HLA-A2 and two were restricted by HLA-A24. Four of the tumor-specific CTL lines lysed autologous fresh tumor cells. Tumor-specific CTLs from two of three patients had cytolytic activity identical with tumor-infiltrating lymphocytes (TIL) derived from tumor biopsies removed earlier and grown in high concentrations (6000 IU/ml) of IL-2. Three of the HLA-A2-restricted tumor-specific CTLs were shown to recognize 293 cells transfected with HLA-A2.1 cDNA and the gene encoding the melanoma Ag, MART-1. In addition, these CTLs recognized the T2 cell line pulsed exogenously with the peptide MART-1(27-35), which is the nine-amino acid immunodominant epitope of the MART-1 Ag recognized on melanoma tumor cells by nearly all HLA-A2-restricted TIL. Thus, we have demonstrated the ability to generate tumor-specific CTLs from PBLs that are similar in their reactivity to TIL. This technique obviates the need for autologous tumor tissue and suggests that PBLs contain sufficient CTL precursors for use in generating antitumor CTLs for cellular immunotherapy trials.
Assuntos
Antígenos de Neoplasias/imunologia , Antígenos HLA-A/imunologia , Melanoma/imunologia , Proteínas de Neoplasias/imunologia , Linfócitos T Citotóxicos/imunologia , Células Tumorais Cultivadas/imunologia , Sequência de Aminoácidos , Testes Imunológicos de Citotoxicidade , Humanos , Imunofenotipagem , Ativação Linfocitária , Teste de Cultura Mista de Linfócitos/métodos , Linfócitos do Interstício Tumoral/imunologia , Antígenos Específicos de Melanoma , Dados de Sequência MolecularRESUMO
Aldose reductase converts glucose to sorbitol, which is further processed to fructose. The enzyme is present in most tissues and its possible physiological role is to produce an electrically neutral, non-diffusible osmolyte in cells exposed to hypertonicity, as typified by the renal medullary cells of the loop of Henlé. The enzyme has a low affinity for glucose, and under normal conditions it processes little substrate. However, in diabetes mellitus, the marked rise in intracellular glucose that occurs in some cells causes marked production of sorbitol. The increased flux and accumulation of sorbitol is damaging to cells and may result in some of the long-term complications of diabetes. In this review, David Tomlinson, Elizabeth Stevens and Lara Diemel discuss the role of aldose reductase and the potential of its inhibitors as therapeutic agents targeted at chronic diabetic complications.
Assuntos
Aldeído Redutase/antagonistas & inibidores , Diabetes Mellitus/terapia , Neuropatias Diabéticas/tratamento farmacológico , Animais , Complicações do Diabetes , Diabetes Mellitus/enzimologia , Nefropatias Diabéticas/tratamento farmacológico , Retinopatia Diabética/tratamento farmacológico , HumanosRESUMO
To evaluate newer therapies for wound infections, it becomes necessary to quantify bacteria that invade from the infected wounds into the adjacent tissues. For example, antibody-targeted photolysis targets the invasive Pseudomonas with antibodies carrying photochemical dyes. A full-thickness burn wound was infected with Pseudomonas aeruginosa with a modification of previous methods. In mice, a skin fold was elevated, and two preheated brass blocks at 92 degrees to 95 degrees C were applied for 5 seconds, producing a 5% total body surface area injury with discrete margins. The eschars were immediately inoculated with Pseudomonas. Survival at 10 days was 100% with burn injury alone and 60% with infected burns. Pseudomonas (10(8)/gm) were recovered from the unburned muscle by 24 hours. The method produced uniform and reproducible quantitative bacteriology within the muscle immediately beneath the burn injury (SL < 0.05). Quantitative comparisons can be used to determine the effectiveness of newer modalities to control Pseudomonas burn wound infections.
Assuntos
Queimaduras/complicações , Infecções por Pseudomonas/microbiologia , Infecção dos Ferimentos/microbiologia , Animais , Queimaduras/mortalidade , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos , Músculos/microbiologia , Infecções por Pseudomonas/complicações , Reprodutibilidade dos Testes , Taxa de Sobrevida , Infecção dos Ferimentos/complicaçõesRESUMO
This study sought to determine the time-course of development of reduced nerve laser Doppler flux in experimental diabetes and the effect on this anomaly of insulin treatment. In addition, we aimed to compare nerve laser Doppler flux in streptozotocin- and genetically-diabetic BB rat models. Sciatic nerve laser Doppler flux in diabetic rats was variable during the 2 days following streptozotocin injection; from day 4, when the measurement was 80% of control, fluxes fell steadily and formed a plateau at 40% of control values after 4 weeks of diabetes. In a second study, using rats with 4-week streptozotocin-diabetes, sciatic nerve laser Doppler flux was reduced to 44% of the value measured in control rats. Treatment of a parallel group of diabetic rats with insulin, by sustained release implants, prevented this ischaemia, so that nerve laser Doppler flux was 91% of controls. Nerve Doppler flux in BB rats with 6-week genetic diabetes was 57% of a control (non-diabetic) BB group. There were no differences in mean arterial pressures between control and diabetic rats in any of the studies. Heart rates of control and insulin-treated diabetic animals were higher than those of the untreated diabetic group; in the other studies heart rates of diabetic animals were numerically lower than controls, but not significantly so. These observations suggest that sciatic nerves of rats with short-term diabetes, whether induced with streptozotocin or of genetic origin, are markedly ischaemic and that this ischaemia in streptozotocin-diabetes is evident within a week of diabetes onset, forms a plateau after 4 weeks and is maintained for at least 2 months.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Diabetes Mellitus Experimental/fisiopatologia , Diabetes Mellitus Tipo 2/fisiopatologia , Insulina/farmacologia , Isquemia/fisiopatologia , Nervo Isquiático/irrigação sanguínea , Nervo Isquiático/fisiopatologia , Animais , Glicemia/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Tipo 1/tratamento farmacológico , Diabetes Mellitus Tipo 1/fisiopatologia , Diabetes Mellitus Tipo 2/tratamento farmacológico , Hemoglobinas Glicadas/metabolismo , Frequência Cardíaca/efeitos dos fármacos , Masculino , Ratos , Ratos Endogâmicos BB , Ratos Wistar , Nervo Isquiático/diagnóstico por imagem , UltrassonografiaRESUMO
Alterations in release of endothelium-derived vasomotor agents could underlie microvascular and neuropathic complications in diabetes. This study examined release of the potent vasodilator prostacyclin, measured as immunoreactive 6-keto prostaglandin F1 alpha, from rat lung, kidney and peripheral nerve. Tissues were taken from control and streptozotocin-diabetic rats which had been treated for 8 weeks with either evening primrose oil (EPO) or, as a control for lipid intake, coconut oil (CO). Lung and kidney slices were incubated in the presence of acetylcholine (ACh), the calcium ionophore 4-Br-A23187, arachidonic acid (AA) or without agonist (basal). Segments of sciatic nerve, with their epineuria punctured, were incubated with or without 4-Br-A23187. Basal prostacyclin release from the lung was significantly higher in rats treated with EPO irrespective of diabetic state (increased by 60% in controls and by 77% in diabetics). Levels were reduced in CO-diabetics compared to EPO-controls (53% reduction) and CO-controls (30% reduction), although this did not reach statistical significance in the latter. Basal prostacyclin release was also significantly reduced in the kidney from CO-diabetics (40% reduction compared to CO-controls and 56% reduction compared to EPO-controls). In the presence of AA, lung prostacyclin release was significantly lower in CO-diabetic rats compared to all other groups (40% reduction compared to EPO-diabetics and 60% compared to both control groups) but there were no differences in renal release between any group. Prostacyclin release by nerves from CO-diabetic rats was significantly reduced (by 91-93%) compared to all other groups.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Diabetes Mellitus Experimental/metabolismo , Epoprostenol/metabolismo , Ácidos Graxos Essenciais/farmacologia , Animais , Glicemia/metabolismo , Peso Corporal , Óleo de Coco , Diabetes Mellitus Experimental/fisiopatologia , Comportamento de Ingestão de Líquido , Comportamento Alimentar , Rim/metabolismo , Ácidos Linoleicos , Pulmão/metabolismo , Masculino , Sistema Nervoso/metabolismo , Oenothera biennis , Óleos de Plantas/farmacologia , Ratos , Ratos Wistar , Ácido gama-LinolênicoRESUMO
1. Alterations in vasoreactivity and endothelial cell function could underlie some of the vascular abnormalities in diabetes. To examine aspects of these phenomena we studied the effects of 4-6 weeks streptozotocin-induced diabetes in the rat on basal and angiotensin II (AII)-stimulated prostacyclin release from isolated lung, perfused at constant flow. In addition, pressure was monitored throughout the lung perfusion as an index of vasomotor tone. 2. The experiment also included lungs from groups of diabetic rats treated with either insulin or an aldose reductase inhibitor (imirestat), to determine whether these treatments influenced the development of any defects seen in untreated diabetes. 3. Despite some indication of a trend towards reduced prostacyclin release in lungs from diabetic rats, neither the basal nor AII-stimulated release was significantly different from that seen in tissues from control animals. There were no significant differences between groups in the average basal perfusion pressure and in either the absolute pressure response to AII or the time of this peak. 4. The area under the perfusion pressure curve during AII infusion was greater in lungs from diabetic animals than in controls indicating a prolonged vasoconstrictor response. This increased pressor response may indicate increased sensitivity of diabetic tissue to AII or a reduced production of vasodilators in response to the vasoconstriction. 5. Whichever mechanism was responsible, this alteration was prevented by insulin treatment but not by aldose reductase inhibition, implicating mechanisms probably unrelated to exaggerated polyol pathway flux.
Assuntos
Aldeído Redutase/antagonistas & inibidores , Diabetes Mellitus Experimental/metabolismo , Epoprostenol/metabolismo , Insulina/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Angiotensina II/farmacologia , Animais , Glicemia/metabolismo , Peso Corporal/efeitos dos fármacos , Metabolismo dos Carboidratos , Endotélio Vascular/citologia , Endotélio Vascular/efeitos dos fármacos , Técnicas In Vitro , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Tono Muscular/efeitos dos fármacos , Perfusão , Ratos , Ratos Wistar , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/metabolismoRESUMO
In rats with 6 weeks streptozotocin-diabetes there was a 53% reduction in sciatic nerve laser Doppler flux compared to controls (p < 0.01). Treatment of a parallel group of diabetic rats with evening primrose oil, by dietary admixture throughout the protocol, prevented this ischaemia (Doppler flux was 91% of evening primrose-oil-treated controls and was not significantly different). There were no differences in systemic arterial pressure. In another experiment evening primrose oil markedly antagonised the development of exaggerated resistance to anoxic conduction failure in sciatic nerves from diabetic rats. The resistance to anoxia of nerves from non-diabetic rats was also reduced by evening primrose oil. These observations suggest that the sciatic nerves of diabetic rats with short-term streptozotocin-diabetes are markedly ischaemic and that this ischaemia is involved in the development of increased resistance to anoxic/ischaemic conduction failure in diabetic nerve. The findings also promote evening primrose oil as a potential treatment to prevent nerve ischaemia.
Assuntos
Diabetes Mellitus Experimental/fisiopatologia , Nefropatias Diabéticas/prevenção & controle , Ácidos Graxos Essenciais/farmacologia , Hipolipemiantes/farmacologia , Isquemia/prevenção & controle , Nervo Isquiático/fisiopatologia , Animais , Glicemia/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Óleo de Coco , Diabetes Mellitus Experimental/sangue , Nefropatias Diabéticas/sangue , Gorduras na Dieta/farmacologia , Ácidos Linoleicos , Masculino , Condução Nervosa/efeitos dos fármacos , Oenothera biennis , Óleos de Plantas/farmacologia , Ratos , Ratos Wistar , Nervo Isquiático/irrigação sanguínea , Nervo Isquiático/efeitos dos fármacos , Ácido gama-LinolênicoRESUMO
This study was designed to determine whether deficient substance P in the sciatic nerve of diabetic rats was associated with a similar reduction in calcitonin gene-related peptide and whether the depletion of either or both peptides could be affected by insulin treatment or by aldose reductase inhibition. Substance P and calcitonin gene-related peptide were measured as immunoreactivities in the same nerve extracts. The sciatic nerve content of substance P was significantly reduced in diabetic rats (0.063 +/- 0.011; all data are mean +/- 1 standard deviation in ng peptide/mg nerve protein; n = 9 for all groups) compared to controls (0.093 +/- 0.026). The calcitonin gene related peptide content was similarly reduced (2.14 +/- 0.49) compared to controls (3.78 +/- 1.21). Tight glycaemic control with insulin prevented completely the deficit in both peptides (substance P = 0.096 +/- 0.021, calcitonin gene-related peptide = 4.66 +/- 0.92). Treatment with the aldose reductase inhibitor, imirestat, corrected the substance P deficit (0.08 +/- 0.018) and attenuated the calcitonin gene-related peptide (3.55 +/- 1.03) depletion seen in the untreated diabetic animals. This indicates that the polyol pathway may play a role in the peptide status of the sciatic nerve. Regression analysis of all data gave r2 = 0.53, indicating a comparable effect of diabetes and the treatments on both peptides.
Assuntos
Aldeído Redutase/antagonistas & inibidores , Peptídeo Relacionado com Gene de Calcitonina/deficiência , Diabetes Mellitus Experimental/metabolismo , Insulina/farmacologia , Nervo Isquiático/metabolismo , Substância P/deficiência , Animais , Glicemia/metabolismo , Fluorenos/farmacologia , Frutose/metabolismo , Glucose/metabolismo , Hidantoínas/farmacologia , Inositol/metabolismo , Masculino , Ratos , Ratos Endogâmicos , Sorbitol/metabolismoRESUMO
Most patients with pancreatic carcinoma are not curable. Surgical palliation of obstructive jaundice and gastric outlet obstruction leaves many patients with severe pain from pancreatic carcinoma. Anesthesiologists have drawn increasing attention to the successful use of postoperative percutaneous celiac plexus block for the treatment of pancreatic pain. Ironically, little attention has been paid to celiac plexus block during laparotomy. We reviewed the cases of 12 patients with pancreatic carcinoma and severe abdominal pain who were treated surgically. All patients had operative celiac plexus block with absolute alcohol at the time of exploratory laparotomy for biliary bypass, gastroenterostomy, or tumor biopsy. Complete postoperative pain relief was obtained in 10 of the 12 patients; two had only partial relief. No operative complications were related to celiac plexus block; one patient died postoperatively of pneumonia. Average postoperative hospital stay was 13 days and average postoperative survival was 3 1/2 months. Most patients had excellent pain relief for at least 2 months or until death. Because most patients treated surgically for pancreatic carcinoma are receiving only palliation with biliary bypass or gastroenterostomy, surgeons should pay increased attention to pain relief. Operative celiac plexus block is easy, safe, and highly effective in relieving the agonizing pain of pancreatic carcinoma.