RESUMEN
Leucine-rich glioma-inactivated (LGI) protein was first thought to have a suppressor effect in the formation of some cancers. Developments in physiology and medicine made it possible to characterize the function of the LGI protein family and its crucial role in different conditions more precisely. These proteins play an important role in synaptic transmission, and dysfunction may cause hyperexcitability. Genetic mutation of LGI1 was confirmed to be the cause of autosomal dominant lateral temporal lobe epilepsy in humans. The LGI2 mutation was identified in benign familial juvenile epilepsy in Lagotto Romagnolo (LR) dogs. Cats with familial spontaneous temporal lobe epilepsy have been reported, and the etiology might be associated with LGI protein family dysfunction. In addition, an autoimmune reaction against LGI1 was detected in humans and cats with limbic encephalitis. These advances prompted a review of LGI protein function and its role in different seizure disorders.
Asunto(s)
Epilepsia/etiología , Proteínas del Tejido Nervioso/fisiología , Animales , Enfermedades de los Gatos/etiología , Enfermedades de los Gatos/genética , Gatos , Enfermedades de los Perros/etiología , Enfermedades de los Perros/genética , Perros , Epilepsia/genética , Epilepsia/veterinaria , Humanos , Péptidos y Proteínas de Señalización Intracelular , Encefalitis Límbica/inmunología , Encefalitis Límbica/veterinaria , Mutación , Proteínas del Tejido Nervioso/genética , Proteínas/genética , Proteínas/inmunología , Proteínas/fisiología , Transmisión Sináptica/fisiologíaRESUMEN
An understanding of the conditions influencing protein binding of catecholamines (CAs) is important in studying their metabolic effects. Unfortunately, reports on plasma protein binding of CAs are scarce, conflicting and mainly performed in vitro. The aim of our in vivo and in vitro studies was to investigate binding and clearance of radioactive adrenaline (epinephrine) ((3)H-A), noradrenaline (norepinephrine) ((3)H-NA) and their metabolites in sheep blood. The time course of the radioactivity in the blood after intravenous injection of (3)H-A and (3)H-NA (3.7 MBq each) in 4 sheep (2 of each sex; total of 8 administrations) was determined. Blood samples were taken from the jugular vein. The highest radioactivity was observed in the first sample (5 min) following injection. Radioactivity showed a biphasic disappearance. An initial stage, in which radioactivity decreased rapidly (within 1 h) after the injection, was followed by a slow stage, lasting for up to 1 month, until background levels were reached. In vitro results indicated that NA and A were present not only in plasma (70%) but also in the erythrocytes (30%; mainly bound to haemoglobin). Sephadex G-25 gel filtration revealed that from the plasma fraction about 15% was strongly bound to proteins (mainly albumin). These results demonstrate that previous experiments in this field have overestimated the percentage of CAs bound to plasma proteins, because binding to haemoglobin was previously not known. In the future, efforts should be made to characterize the adduct products of CAs and establish an assay to determine them in vivo. If this could be achieved, it would yield a valuable tool for measuring the stress experienced for a longer period.
Asunto(s)
Epinefrina/sangre , Epinefrina/metabolismo , Norepinefrina/sangre , Norepinefrina/metabolismo , Ovinos/sangre , Albúminas , Animales , Epinefrina/química , Femenino , Masculino , Norepinefrina/química , TritioRESUMEN
Sera of healthy pregnant (group I, n = 11) and non-pregnant (group II, n = 11) bitches were screened for autoantibodies (AAb). In both groups, blood samples were drawn every fifth day between days 5 and 55 after mating. Serum was analysed via indirect immunofluorescence (IIF) with the Canine ANA HEp-2 Screening Kit. In all animals, anticytoplasmic AAb were detected. Utilizing primate-heart substrate slides AAb against contractile proteins of the cytoplasm could be observed. The predominating fluorescence pattern in pregnant animals resembled above all desmin, which was proven via Western blot. The sera were then pre-incubated with tropomyosin, actin, vimentin, vinculin and keratin solutions, and assessed on HEp-2 slides and on human and canine fibroblasts as well. The latter substrate was used to verify whether the detected Ab were in fact AAb. Utilizing tropomyosin, revealed elimination of the cytoplasmic fluorescences on all three substrates. It is therefore assumed, that in sera of healthy dogs, AAb against contractile structure proteins of the cytoplasm are present regularly. The majority of pregnant bitches presented with higher end-point titres (EPT), than to be found in non-pregnant dogs. AAb against desmin played the key role in those patterns. In addition, sera were screened for thyroid specific AAb, namely thyroglobulin, thyroid peroxidase (TPO), T3 and T4, and for AAb against insulin by ELISA or Western blot (TPO). Only in two of the pregnant bitches a weak positive reaction (1:100) for T3-AAb was detected.
Asunto(s)
Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Perros/sangre , Factores Inmunológicos/sangre , Preñez/sangre , Animales , Perros/inmunología , Electroforesis en Gel de Poliacrilamida/métodos , Electroforesis en Gel de Poliacrilamida/veterinaria , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Factores Inmunológicos/inmunología , Peso Molecular , Embarazo , Preñez/inmunología , Estudios SeroepidemiológicosRESUMEN
The aim of this study was to investigate, whether the activity of matrix metalloproteinase (MMP)-2 and -9 in the serum of pregnant and non-pregnant bitches differs significantly. For this purpose, 81 blood samples were taken from pregnant bitches at days 5-13, 15-21, 24-31, 34-40 and 41-50 after mating, and 51 samples from non-pregnant animals at corresponding times. Relative enzyme activity, calculated as the percentage of serum enzyme activity on enzyme activity in a control sample, was determined with a commercially available assay after activation of serum MMPs with p-aminophenylmercuric acetate (APMA). In addition, serum oestradiol (E(2)) and progesterone (P(4)) concentrations were measured with an enzymeimmunoassay (EIA). In the pregnant bitches, at days 5-13 and 15-21 after mating, the mean activity of both MMPs was significantly higher than in non-pregnant animals (28.5% vs 24.5% and 27.7% vs 22.6%; p < 0.01). Moreover, in the pregnant bitches, significant correlations were detected between the serum enzyme activity and the serum concentrations of E(2) (-0.900; p < 0.05) and P(4) (+0.667; p < 0.05).
Asunto(s)
Estro/sangre , Metaloproteinasa 2 de la Matriz/sangre , Metaloproteinasa 9 de la Matriz/sangre , Preñez/sangre , Animales , Estudios de Casos y Controles , Perros , Femenino , EmbarazoRESUMEN
The thyroglobulin of a ram of the East Friesian milk sheep breed suffering from goitre was investigated by physico- and immunochemical methods. The respective ram was the only animal amongst the other sheep of the flock, that exhibited severe goitre, additionally showing depressed behaviour. Results of the thyroid-stimulating hormone response test were indicative of hypothyroidism. The dysfunction of the thyroid gland could be treated by additional iodine supplementation quite successfully, although all sheep had been given iodinated cattle salt throughout the course of the history. Without reducing conditions sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of isolated thyroglobulin molecules of the ram and control sheep did not reveal different band patterns, but under reducing conditions different band patterns were evident for the respective animals: the ram's thyroglobulin displayed two main bands, those of healthy reference sheep only one. Both bands reacted equally with anti-thyroglobulin antibodies, even with those produced by immunizing rabbits with single bands. The reduced single thyroglobulin band of healthy sheep corresponded to a truncated form of that molecule, whereas the additional main band of the ram was a more resistant, intact thyroglobulin subunit, as was shown by mass spectrometry. In conclusion, results of physico- and immunochemical investigations gave evidence of a modification of thyroglobulin with suspected different iodine binding properties in the ram. The latter finding may have clinical relevance in similar cases in other species, as it is an example of the impact that a minor change in a protein molecule may have on a complete metabolic pathway. Additionally, it could be shown, that in the ovine species the generally found single main band of thyroglobulin after reduction is a truncated form and not an intact subunit. This truncation seems to be induced in vitro by the reductive sample pretreatment prior to SDS-PAGE.
Asunto(s)
Bocio/veterinaria , Hipotiroidismo/veterinaria , Enfermedades de las Ovejas/diagnóstico , Tiroglobulina/metabolismo , Animales , Diagnóstico Diferencial , Electroforesis en Gel de Poliacrilamida/veterinaria , Bocio/complicaciones , Bocio/diagnóstico , Hipotiroidismo/complicaciones , Hipotiroidismo/diagnóstico , Masculino , Ovinos , Enfermedades de las Ovejas/patología , Pruebas de Función de la Tiroides/veterinariaRESUMEN
Enzyme immunoassays (EIAs) for the determination of autoanti-bodies (AA) to thyroid antigens in canine serum were developed. Streptavidin (SA) was immobilized as capture molecule on microtitreplates (MTP). Thyroglobulin (Tg) purified from canine thyroids and the thyroid hormones thyroxine and triiodothyronine (T3 and T4) were conjugated to biotin labelling reagents and attached to the MTP over the SA-biotin bridge. Bound AA were detected with anti-dog-immunoglobulin G (IgG) labelled with horseradish peroxidase. Serum samples from dogs which were allotted to four groups were analysed: A (n = 31), biochemical evidence of hypothyroidism; B (n = 76), clinical signs of hypothyroidism; C (n = 47), euthyroid with non-thyroidal disease; D (n = 186), clinically healthy. The validity of the assays was tested with two different methods. After thiophilic absorption chromatography of positive sera, a positive reaction in the EIA was only detected in those fractions which coeluted with the canine IgG standard. Furthermore, the positive reaction was blocked by the addition of the corresponding antigen. In 55% of the hypothyroid dogs AA to Tg and/or T3 and T4, respectively, were found (up to a titre of 1 : 1600). In group B 34% of the dogs were diagnosed positive, but the titre was lower (up to 1 : 400). In the groups C and D the number of dogs with AA and their titre was significantly lower. Two different methods for distinguishing positive and negative test results were compared in order to increase the specificity of the tests without decreasing the sensitivity. The EIAs are precise and based on high agreement with previous reported assays able to discriminate dogs with thyroiditis from healthy ones. These assays represent a good alternative to the isotope assays generally used for the analysis of AA to T4 and T3.
Asunto(s)
Autoanticuerpos/sangre , Enfermedades de los Perros/diagnóstico , Perros/inmunología , Hipotiroidismo/veterinaria , Técnicas para Inmunoenzimas/veterinaria , Animales , Estudios de Casos y Controles , Enfermedades de los Perros/sangre , Enfermedades de los Perros/inmunología , Perros/sangre , Femenino , Hipotiroidismo/diagnóstico , Técnicas para Inmunoenzimas/normas , Masculino , Valor Predictivo de las Pruebas , Reproducibilidad de los Resultados , Tiroglobulina/sangre , Tiroglobulina/inmunología , Tiroxina/sangre , Tiroxina/inmunología , Triyodotironina/sangre , Triyodotironina/inmunologíaRESUMEN
Oestrogen and progestagen metabolites were measured in the faeces of five female giant anteaters (Myrmecophaga tridactyla), to characterise the oestrous cycle and pregnancy. Faecal samples were collected twice weekly for a minimum of 6 months, and immunoreactive progestagens and oestrogens were analysed using enzyme immunoassays (EIA). For progestagens, two antibodies that cross-reacted with 20alpha-hydroxy- or 20-oxo-progestagens were used. Both assays effectively monitored ovarian cyclicity; however, the concentrations obtained using the antibody for 20alpha-hydroxy-progestagens were higher, and the hormonal changes were more pronounced. Regular ovarian cycles were identified in three of the five females. Average (+/-SEM) length of the oestrous cycle (n=10) was 51.4+/-5.6 days. Peak concentrations of 20alpha-hydroxy-progestagens ranged from 80-660 ng/g of faeces and those of oestrogens from 20-100 ng/g. Hormone concentrations were measured during parts of two pregnancies and during four post-partum periods. The length of one gestation (from oestrous oestrogen peak until parturition) was 184 days. In the second half of gestation, progestagen concentration started to increase above luteal phase values; in the week before parturition it was approximately 20 times higher than those during the luteal phase. Concentrations of excreted oestrogens began to increase after two thirds of gestation and exceeded that of the follicular phase by approximately 2.5-fold in the week before parturition. Onset of ovarian cyclicity after parturition varied from 4-11 weeks. In conclusion, the measurement of faecal immunoreactive progestagens and oestrogens in the giant anteater indicated an ovarian cycle of approximately 7 weeks in length and provided potentially useful data for successful breeding management.