Preservation and Immunogold Localization of Lipids by Freeze-Substitution and Low Temperature Embedding

The success of postembedding immunocytochemistry depends largely on the preparation methods. The requirements for structural preservation and immunocytochemistry are in some cases contradictory. This is especially the case in the study of lipid-rich structures and the localization of lipid components. Earlier work on freeze-substitution has shown that this method is very promising for the preservation of lipids and the immunocytochemical localization of lipids at the electron microscopical level. In this study we show that freeze-substitution in combination with low temperature embedding in Lowicryl HM20 has fulfilled this promise. Lamellar bodies in alveolar type II cells contain about 90% lipids and are very difficult to preserve in ultrathin cryosections. Lowicryl sections of freeze-substituted lung tissue shows excellent preservation of lamellar bodies in combination with immunogold localization of a hydro- phobic surfactant protein. With an antibody against the Forssman glycolipid we demonstrate a highly reproducible intracellular localization of this glycolipid with high specificity and resolution. This method results in the retention of lipids and glycolipids and allows postembedding immunogold labeling

Поняття “леґітимність” та “леґітимація” як теоретичні інновації М.Вебера: зміст і роль у сучасній соціології

Forssman antigen, a neutral glycosphingolipid carrying five monosaccharides, was localized in epithelial MDCK cells by the immunogold technique. Labeling with a well defined mAb and protein A-gold after freeze-substitution and low temperature embedding in Lowicryl HM20 of aldehyde-fixed and cryoprotected cells, resulted in high levels of specific labeling and excellent retention of cellular ultrastructure compared to ultra-thin cryosections. No Forssman glycolipid was lost from the cells during freeze-substitution as measured by radio-immunostaining of lipid extracts. Redistribution of the glycolipid between membranes did not occur. Forssman glycolipid, abundantly expressed on the surface of MDCK II cells, did not move to neighboring cell surfaces in cocultures with Forssman negative MDCK I cells, even though they were connected by tight junctions. The labeling density on the apical plasma membrane was 1.4-1.6 times higher than basolateral. Roughly two-thirds of the gold particles were found intracellularly. The Golgi complex was labeled for Forssman as were endosomes, identified by endocytosed albumin-gold, and lysosomes, defined by double labeling for cathepsin D. In most cases, the nuclear envelope was Forssman positive, but the labeling density was 10-fold less than on the plasma membrane. Mitochondria and peroxisomes, the latter identified by catalase, remained free of label, consistent with the notion that they do not receive transport vesicles carrying glycosphingolipids. The present method of lipid immunolabeling holds great potential for the localization of other antigenic lipids

Довгостроковий часовий прогноз розвитку карсту на Передкарпатті

Здійснено довгостроковий часовий прогноз активізації сульфатного карсту в Передкарпатті. В якості факторів активізації розглянуті сонячна і сейсмічна активність, метеорологічні та гідрогеологічні дані. Основою часового прогнозу, здійсненого до 2020 року, служить теорія про ритмічність природних явищ, їх взаємозв’язок і взаємообумовленість.Осуществлен временной прогноз активизации сульфатного карста в Предкарпатье. В качестве факторов активизации рассмотрены солнечная и сейсмическая активность, метеорологические и гидрогеологические данные. Основой временного прогноза, осуществленного до 2020 года является теория о ритмичности природных явлений, их взаимосвязи и взаимообусловленности.The long-term time forecast of sulfate cockpit activation throughout the Piedmont Carpathians has been accomplished. The leading factors-activators include: solar and seismic activity as well as meteorological and hydro-geological data. The backbone of this prognosis for the period up to 2020 is represented by the theory of rhythm among natural phenomena, their interconnection and stipulation

Принцип коеволюції в контексті ноосферної концепції В.Вернадського

Досліджується принцип коеволюції людини та природи в контексті ноосферного вчення В.Вернадського. Аналізуються особливості світогляду суспільства ноосферної цивілізації.Исследуется принцип коэволюции человека и природы в контексте ноосферного учения В.Вернадского. Анализуются особенности мировоззрения общества ноосферной цивилизации.The principle of coevolution of person and nature is investigated in the context of noospherian doctrine of V.Vernadsky. The particularities of world view оf society of noospherian civilization are analyzed

Radiographic and computed tomographic assessment of the development of the antebrachia and elbow joints in Labrador Retrievers with and without medial coronoid disease

Objectives: To compare the development, monitored by radiography and computed tomography, of the antebrachia and elbow joints in seven Labrador Retrievers with healthy elbow joints and in seven Labrador Retrievers that developed medial coronoid disease (MCD), in order to determine whether disturbances in the development of the antebrachia and elbow joints, between the age of six and 17 weeks may lead to medial coronoid disease. Methods: A prospective study of 14 Labrador Retrievers in their active growth stage was performed. The development of the antebrachia and elbow joints was assessed between six and 17 weeks of age using radiography and computed tomography determining the development of secondary ossification centres, radioulnar length ratio, radial angulation, and inter-relationship between the humerus, ulna and radius. Results: For the parameters of ossification of secondary ossification centres, radioulnar length ratio, radial angulation, and joint congruence evaluation, there was no significant difference in the development of the antebrachia and elbow joints of seven Labrador Retrievers positive and seven Labrador Retrievers negative for MCD at the age of six to 17 weeks. Clinical significance: These findings demonstrate that the development of MCD in the Labrador Retrievers in our study was not related to any disturbance in the development of the antebrachia and elbow joints during the rapid growth phase

Phenotypic characteristics of Hydrocephalus in stillborn Friesian foals

Hydrocephalus is uncommon in horses. However, in recent years, it has become clear that the prevalence of hydrocephalus is greater in Friesian horses than in other breeds probably due to their limited gene pool. Before identification of candidate genes that predispose to the development of hydrocephalus in Friesian horses can be pursued, an in-depth, phenotypic, pathological description of the condition in Friesians would be of great benefit. Our study aimed to characterize the morphology of hydrocephalus in Friesian horses, to support further investigation of the genetic background of this condition. Four stillborn Friesian foals with hydrocephalus were examined macroscopically and microscopically and compared with 2 normal stillborn Friesian foals without hydrocephalus. In all clinical cases, tetraventricular and venous dilatations were observed, together with malformation of the petrosal bone and, as a result, narrowing of the jugular foramen. These observations suggest a communicative hydrocephalus with a diminished absorption of cerebrospinal fluid into the systemic circulation at the venous sinuses due to a distorted, nonfunctional jugular foramen. This type of hydrocephalus is also recognized in humans and dogs and has been linked genetically to chondrodysplasia; this has already been recognized in dwarfism, which is another monogenetic defect in Friesian horses.http://vet.sagepub.com/am201

The early development of medial coronoid disease in growing Labrador retrievers: radiographic, computed tomographic, necropsy and micro-computed tomographic findings

Medial coronoid disease (MCD) encompasses lesions of the entire medial coronoid process (MCP), both of the articular cartilage and the subchondral bone. To detect the earliest signs of MCD, radiography and computed tomography were used to monitor the development of MCD in 14 Labrador retrievers, from 6 to 7 weeks of age until euthanasia. The definitive diagnosis of MCD was based on necropsy and micro-computed tomography findings. The frequency of MCD in the dogs studied was 50%. Radiographic findings did not provide evidence of MCD, ulnar subtrochlear sclerosis or blunting of the cranial edge of the MCP. Computed tomography was more sensitive (30.8%) than radiography (0%) in detecting early MCD, with the earliest signs detectable at 14 weeks of age. A combination of the necropsy and micro-computed tomography findings of the MCP showed that MCD was manifested as a lesion of only the subchondral bone in dogs <18 weeks of age. In all dogs (affected and unaffected), there was close contact between the base of the MCP and the proximal radial head in the congruent joints. Computed tomography and micro-computed tomography findings indicated that the lesions of MCD probably originated at the base of the MCP