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1.
J Dairy Sci ; 88(8): 2774-82, 2005 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16027191

RESUMO

Histopathology of the dermo-epidermal junction in the lamellar region of front claws was examined in 6 dairy heifers given an alimentary oligofructose overload and compared with sections from a control group of 6 heifers. Four of the 6 heifers administered oligofructose developed clinical signs of acute laminitis before they were euthanized. Postmortem samples from front claws were processed for histology. Eleven histopathologic characteristics were selected from the existing literature and used in a blinded evaluation of sections. In total, 104 front claw samples, including 8 samples from 2 cows having spontaneously occurring acute laminitis, were evaluated histologically using hematoxylin and eosin as well as periodic acid-Schiff staining. The major morphological features associated with oligofructose-induced acute clinical laminitis were stretching of lamellae, dermal edema, hemorrhage, changes in basal cell morphology, presence of white blood cells in dermis, and signs of basement membrane detachment. Changes at the lamellar junction of claw tissue affected by oligofructose-induced clinical laminitis resembled tissue from the 2 cows suffering from spontaneous acute clinical laminitis, and generally were consistent with existing descriptions of laminitis histopathology. Important exceptions to existing descriptions in the literature were stretching of lamellae and basement membrane changes. Not previously described, we considered these early signs of acute laminitis. In conclusion, this study documents that oligofructose-induced clinical laminitis is associated with histopathological changes at the lamellar interface. A weakened dermo-epidermal junction is a possible intermediate stage in the pathophysiology of bovine sole ulceration at the typical site.


Assuntos
Doenças dos Bovinos/induzido quimicamente , Doenças dos Bovinos/patologia , Doenças do Pé/veterinária , Oligossacarídeos/administração & dosagem , Animais , Membrana Basal/patologia , Bovinos , Corantes , Amarelo de Eosina-(YS) , Epiderme/patologia , Feminino , Corantes Fluorescentes , Doenças do Pé/induzido quimicamente , Doenças do Pé/patologia , Hematoxilina , Casco e Garras/patologia , Inflamação/induzido quimicamente , Inflamação/patologia , Inflamação/veterinária , Coloração e Rotulagem
2.
J Dairy Sci ; 87(6): 1747-50, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15453488

RESUMO

The objectives of this study were to compare analytical instruments used in independent laboratories to measure milk urea nitrogen (MUN) and determine whether any components in milk affect the recovery of MUN. Milk samples were collected from 100 Holstein cows fed one ration in a commercial dairy herd with a rolling herd average of 9500 kg. Half of each sample was spiked with 4 mg/dL of urea N, while the other half was not, to determine recovery. Both milk samples (spiked and not spiked) were sent to 14 independent laboratories involved in the MUN Quality Control Program through National Dairy Herd Improvement Association and analyzed for MUN, fat, protein, lactose, somatic cell count (SCC), and total solids. The laboratories analyzed MUN using CL-10 (n = 3), Skalar (n = 2), Bentley (n = 3), Foss 4000 (n = 3) or Foss 6000 (n = 3) systems. When recovery of MUN was evaluated among the 5 analytical methods, the mean recoveries for the Bentley, Foss 6000, and Skalar systems were 92.1 (SE = 2.76%), 95.4 (SE = 10.1%), and 95.1% (SE = 7.61%), respectively, and did not differ from each other. However, MUN recovery was 85.0% (SE = 2.8%) for the CL-10 system and 47.1% (SE = 9.9%) for the Foss 4000 system, both of which differed from the other 3 systems. Recoveries from Foss 4000, Foss 6000, and Skalar varied among laboratories using the same instrument. As initial MUN concentration increased, recovery decreased using the Bentley and CL-10 systems. Increasing milk fat resulted in a decrease in recovery using the Foss 6000 system. For 4 of the 5 methods, recovery of MUN was not associated with specific milk components. Recovery of MUN was inconsistent for laboratories using the Foss 4000 and the Foss 6000 method and using these systems may result in an overestimation or underestimation of MUN.


Assuntos
Indústria de Laticínios/métodos , Leite/química , Nitrogênio/análise , Ureia/análise , Animais , Bovinos , Contagem de Células/veterinária , Feminino , Lactação , Lipídeos/análise , Proteínas do Leite/análise , Controle de Qualidade , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
3.
J Dairy Sci ; 87(6): 1848-53, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15453501

RESUMO

The objective of this study was to compare the instruments and laboratories that are currently used for analysis of milk urea nitrogen (MUN) for bulk-tank milk samples. Two replicate samples from each bulk tank on 10 different dairy farms were sent to 12 Dairy Herd Improvement Association (DHIA) laboratories throughout the US for MUN analysis. Two laboratories used 2 different methods for MUN analysis for a total of 14 analyses on 20 samples (n = 280). Values of MUN were analyzed using a random effects model with farm, laboratory, and farm x laboratory variance components. Greater than 98% of the variance in measured MUN was attributed to farm-to-farm variance for analysis of MUN by the Bentley, CL 10, Foss 6000, and Skalar instruments. However, for the laboratories using the Foss 4000 system, <60% of the variance in MUN was attributed to farm-to-farm variance. Laboratories using the Bentley, CL 10, Foss 6000, and Skalar instruments provided slightly different results for MUN analysis, but >95% of sample measurements fell within 1.75 mg/ dL of each other. The laboratories using Foss 4000 differed from each other, and 95% of samples fell within 5 mg/dL of the CL 10 measurement. Laboratories using the Foss 4000 instrument did not consistently provide measurements of MUN that were similar to each other or to the measurements of the other instruments.


Assuntos
Laboratórios/normas , Leite/química , Nitrogênio/análise , Ureia/análise , Animais , Bovinos , Indústria de Laticínios/instrumentação , Indústria de Laticínios/métodos , Feminino , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
4.
Equine Vet J ; 36(3): 230-5, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15147130

RESUMO

REASONS FOR PERFORMING STUDY: Light microscopical studies show that the key lesion of laminitis is separation at the hoof lamellar dermal-epidermal interface. More precise knowledge of the damage occurring in the lamellar basement membrane zone may result if laminitis affected tissue is examined with the transmission electron microscope. This could lead to better understanding of the pathogenesis of lesions and the means of treatment or prevention. OBJECTIVES: To investigate the ultrastructure of acute laminitis as disease of greater severity is induced by increasing oligofructose (OF) dosage. METHODS: Three pairs of normal horses, dosed with OF at 7.5, 10 and 12.5 g/kg bwt via nasogastric intubation, developed laminitis 48 h later. Following euthanasia, their forefeet were processed for transmission electron microscopy. Lamellar basal cell hemidesmosome (HD) numbers and the distance between the basal cell plasmalemma and the lamina densa of the basement membrane were estimated and compared to control tissue. RESULTS: Increasing OF dosage caused greater HD loss and more severe laminitis. The characteristic separation of the basement membrane, cytoskeleton failure and rounded basal cell nuclei results from combined HD dysassembly and anchoring filament failure. CONCLUSIONS: Without properly assembled HDs, dysadhesion between the lamina densa of the basement membrane (BM) and epidermal basal cells occurs, emphasising the fundamental importance of HDs in maintaining attachment at the lamellar interface. Medical conditions that trigger lamellar matrix metalloproteinase (MMP) activation and/or compromise entry of glucose into lamellar basal cells appear to promote loss and failure of HDs and, therefore, laminitis development. POTENTIAL RELEVANCE: A correlation between lameness severity and escalating loss of lamellar HDs now exists. Therapy aimed at protecting the lamellar environment from haematogenous delivery of MMP activators or from glucose deprivation may control laminitis development.


Assuntos
Doenças do Pé/veterinária , Hemidesmossomos/ultraestrutura , Casco e Garras/ultraestrutura , Doenças dos Cavalos/patologia , Oligossacarídeos/farmacologia , Animais , Membrana Basal/citologia , Membrana Basal/efeitos dos fármacos , Membrana Basal/ultraestrutura , Relação Dose-Resposta a Droga , Feminino , Doenças do Pé/induzido quimicamente , Doenças do Pé/patologia , Hemidesmossomos/efeitos dos fármacos , Casco e Garras/efeitos dos fármacos , Casco e Garras/patologia , Doenças dos Cavalos/induzido quimicamente , Cavalos , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Microscopia Eletrônica de Transmissão/métodos , Microscopia Eletrônica de Transmissão/veterinária , Oligossacarídeos/efeitos adversos , Distribuição Aleatória , Índice de Gravidade de Doença
5.
Equine Vet J ; 36(3): 242-7, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15147132

RESUMO

REASONS FOR PERFORMING STUDY: The key lesion of laminitis is separation at the hoof lamellar dermal-epidermal interface. For this to happen the structural and adhesion proteins of the basement membrane zone must be altered. Which proteins and how damage to them leads to the lamellar separation of laminitis is unknown. OBJECTIVES: To investigate lamellar hemidesmosome and cytoskeleton damage and basement membrane dysadhesion using light microscopy (LM) and immunofluorescence microscopy (IFM). METHODS: Cryostat sections of lamellar tissues from 2 control and 6 Standardbred horses with oligofructose induced laminitis were studied using LM and IFM. Plectin, integrin alpha6 and BP230 antibody was used to label hemidesmosome intracellular plaque proteins and anti-BP180 and anti-laminin 5 (L5) was used to label anchoring filament (AF) proteins. Cytoskeleton intermediate filaments were labelled using anti-cytokeratin 14. The primary antibodies of selected sections were double labelled to show protein co-localisation. RESULTS: Laminitis caused reduction of transmembrane integrin alpha6, the AF proteins BP180 and L5, and failure of co-localisation of BP180 and L5. Proteins of the inner hemidesmosomal plaque, plectin and BP230, were unaffected. CONCLUSIONS: Loss of co-localisation of L5 and BP180 suggests that, during the acute phase of laminitis, L5 is cleaved and therefore, the AFs connecting the epidermis to the dermis, fail. Without a full complement of AFs separation at the lamellar dermo-epidermal junction occurs. POTENTIAL RELEVANCE: Suppressing or inhibiting metalloproteinase activity may prevent L5 cleavage and therefore the lamellar dermo-epidermal separation of laminitis.


Assuntos
Doenças do Pé/veterinária , Casco e Garras/patologia , Doenças dos Cavalos/patologia , Laminina/fisiologia , Doença Aguda , Animais , Anticorpos/imunologia , Membrana Basal/citologia , Membrana Basal/patologia , Adesão Celular/fisiologia , Citoesqueleto/patologia , Doenças do Pé/patologia , Hemidesmossomos/patologia , Casco e Garras/citologia , Cavalos , Imuno-Histoquímica/veterinária , Inflamação/patologia , Inflamação/veterinária , Laminina/análise , Microscopia de Fluorescência/veterinária
6.
Equine Vet J ; 36(3): 261-6, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15147135

RESUMO

REASONS FOR PERFORMING STUDY: Acute laminitis is characterised by hoof lamellar dermal-epidermal separation at the basement membrane (BM) zone. Hoof lamellar explants cultured in vitro can also be made to separate at the basement membrane zone and investigating how this occurs may give insight into the poorly understood pathophysiology of laminitis. OBJECTIVES: To investigate why glucose deprivation and metalloproteinase (MMP) activation in cultured lamellar explants leads to dermo-epidermal separation. METHODS: Explants, cultured without glucose or with the MMP activator p-amino-phenol-mercuric acetate (APMA), were subjected to tension and processed for transmission electron microscopy (TEM). RESULTS: Without glucose, or with APMA, explants under tension separated at the dermo-epidermal junction. This in vitro separation occurred via 2 different ultrastructural processes. Lack of glucose reduced hemidesmosomes (HDs) numbers until they disappeared and the basal cell cytoskeleton collapsed. Anchoring filaments (AFs), connecting the basal cell plasmalemma to the BM, were unaffected although they failed under tension. APMA activation of constituent lamellar MMPs did not affect HDs but caused AFs to disappear, also leading to dermo-epidermal separation under tension. CONCLUSIONS: Natural laminitis may occur in situations where glucose uptake by lamellar basal cells is compromised (e.g. equine Cushing's disease, obesity, hyperlipaemia, ischaemia and septicaemia) or when lamellar MMPs are activated (alimentary carbohydrate overload). POTENTIAL RELEVANCE: Therapies designed to facilitate peripheral glucose uptake and inhibit lamellar MMP activation may prevent or ameliorate laminitis.


Assuntos
Doenças do Pé/veterinária , Glucose/administração & dosagem , Casco e Garras/metabolismo , Doenças dos Cavalos/metabolismo , Metaloproteinases da Matriz/metabolismo , Acetato de Fenilmercúrio/análogos & derivados , Doença Aguda , Animais , Membrana Basal/metabolismo , Membrana Basal/patologia , Membrana Basal/ultraestrutura , Técnicas de Cultura/veterinária , Ativação Enzimática , Epiderme/metabolismo , Epiderme/patologia , Doenças do Pé/enzimologia , Doenças do Pé/metabolismo , Doenças do Pé/patologia , Glucose/metabolismo , Casco e Garras/patologia , Casco e Garras/ultraestrutura , Doenças dos Cavalos/enzimologia , Doenças dos Cavalos/patologia , Cavalos , Inflamação/enzimologia , Inflamação/metabolismo , Inflamação/patologia , Inflamação/veterinária , Microscopia Eletrônica de Transmissão/veterinária , Acetato de Fenilmercúrio/farmacologia
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