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Biomedical subjects

M Kraemer

Publications and source records attributed to M Kraemer.

At least 91 records · Page 5Linked to original sources

The effect of dexamethasone on transferrin secretion by cultured fetal rat hepatocytes.

Cultured fetal rat hepatocytes derived from 12, 15 and 19-day gestation rats are capable of secreting transferrin. When dexamethasone is added to the medium an increased secretion rate is observed. The changes in secretion rates in control as well as dexamethasone-treated cells during culture have been shown to correlate with the level of mRNA coding for transferrin. Immunocytochemical experiments show that initially all hepatocytes contain transferrin which is localized in the lumina of the perinuclear space, rough endoplasmic reticulum and in the saccules and vesicles of the Golgi apparatus. During culture, particularly in control cells, the intensity of labelling varies from cell to cell. In addition, adjacent cells are observed to label more intensely in different intracellular organelles.

Animals↗

The effect of dexamethasone on albumin production by fetal rat hepatocytes in culture.

Hepatocytes derived from 15 and 19-day gestation rats synthesize and secrete albumin during culture. Albumin secretion is maintained when the culture medium is supplemented with dexamethasone but declines in its absence. The fall in secretion rate correlates with the level of albumin messenger RNA in the respective cultures. Even when dexamethasone is present, the level of albumin production in 19-day gestation hepatocytes is 6 to 7 times greater than that observed in hepatocytes derived from 15-day gestation rats. Immunocytochemical studies were undertaken to establish whether the difference in secretion rate was due to a difference in the amount of albumin produced by all the hepatocytes of the respective cultures or whether there were fewer hepatocytes which were capable of synthesizing albumin in the less mature liver. The results indicate that albumin production is reduced in all hepatocytes when cultured in the absence of dexamethasone.

Albumins↗

Ultrastructural indirect immunolocalization of transferrin in cultured rat hepatocytes permeabilized with saponin.

Transferrin was localized in 48-hr cultured adult rat hepatocytes by indirect immunoperoxidase following paraformaldehyde--glutaraldehyde fixation and the use of saponin as a membrane permeabilizing agent. The protein, present in all the parenchymal cells in variable amounts, was found to be specifically located in the endoplasmic reticulum and Golgi apparatus. These results are consistent with recent reports claiming that all adult hepatocytes may synthesize a given liver plasma protein at a given time. The procedure used in this study should be particularly useful for the detection of intracellular antigens in various intact cell types.

Animals↗

The influence of mode of fixation, type of fixative and vehicles on the same rat liver: a morphometric/stereologic study by light and electron microscopy.

The influence on cellular and subcellular structure was studied by comparing perfusion versus immersion fixation with cacodylate buffered glutaraldehyde, osmium or glutaraldehyde immersion fixation with two vehicles, phosphate and cacodylate buffer. The study was performed in the same animal using the left liver lobe for the four immersion fixation experiments and the remaining lobes for perfusion. The preparative steps were done under identical conditions in one operation. The mode of fixation was the most influential factor with clear (15%) increases for the volume of hepatocytes (mainly their cytoplasm), and the mitochondria (30%), when comparing immersion to perfusion fixation. This mitochondrial enlargement was particularly displayed in the organelles' average profile area, being more than doubled (211%) in immersion fixation. Similarly high profile enlargements (150%) were also found after osmium immersion fixation. Changes in nuclei were minor compared to the cytoplasm. There was a good correlation between the volume changes of the hepatocyte cytoplasm and the mitochondria (r = 0.85, p less than 0.01). Qualitative results revealed that there exists a zone of good electron microscopic preservation 2-3 cell layers beneath the block edge in immersion fixed material, whereas perfusion fixed tissue displays a homogeneous fine structure.

Animals↗

[Effect of fixation on immunolocalization of transferrin and albumin in the liver of the adult rat].

The influence of the fixation procedure on the localization of albumin and transferrin in adult rat liver has been carried out using an indirect immunoperoxidase technique at the light and electron microscopic levels. Perfusion and immersion fixations with different concentrations of paraformaldehyde (with or without addition of glutaraldehyde) have been investigated. According to the mode of fixation (perfusion versus immersion) and the concentration of the fixative, the number of albumin and transferrin containing hepatocytes could vary from 10% to 100%, and different labeling patterns could be observed at the electron microscopic level. For the same concentration of fixative, a perfusion fixation induces a less intense labeling than an immersion fixation. Thus similar results are obtained after immersion fixation in 6% paraformaldehyde + 0.25% glutaraldehyde or after perfusion fixation in 4% paraformaldehyde + 0.025% glutaraldehyde. Similar data are noticed after immersion fixation in 4% paraformaldehyde or after perfusion fixation in 1% paraformaldehyde + 0.025% glutaraldehyde. Moreover, perfusion fixation induced a more fine cell structure preservation than immersion fixations and avoided the appearance of zones of fixation.

Albumins↗

Amino acid sequence of crayfish (Astacus fluviatilis) trypsin If.

The complete amino acid sequence of trypsin from the crayfish Astacus fluviatilis has been determined. The protein was fragmented with cyanogen bromide after S-carboxymethylation of the reduced disulfide bonds and by trypsin after S-carboxymethylation as well as after succinylation of lysine residues and aminoethylation of the reduced disulfide bonds. Peptides were purified by gel filtration and by reversed-phase high-performance liquid chromatography. Stepwise degradation was performed in a spinning cup sequencer. The enzyme contains 237 amino acid residues and has a molecular weight of 25 030. In contrast to bovine trypsin, it contains three rather than six disulfide bonds which are paired in the same fashion as those in trypsin from Streptomyces griseus. The constituents of the active site of bovine trypsin are present in corresponding positions in the crayfish enzyme. Crayfish trypsin shows 43.6% sequence identity with the bovine enzyme as compared to 40.0% identity with the S. griseus enzyme. The present analysis affords the first detailed view into the evolution of trypsins at the invertebrate level.

Amino Acid Sequence↗

[Effect of paraformaldehyde fixation on the qualitative preservation and stereological parameters of the adult rat liver].

Studies were made to compare the impact of immersion fixations with different concentrations of paraformaldehyde (10%, 6% + 0,25% glutaraldehyde, 4% and 1%) on the cellular and subcellular structure preservation. The study was performed on 3 animals. Similar conditions existed for all the preparative steps, since they were done in one operation. Cellular parameters as the volume densities of hepatocytes (VVH), nuclei (VVNH), cytoplasm (VVCYT) and extra-hepatocytic space (VVEX) were assessed by light microscopy on toluidine blue stained semi-thin sections at X 1 000. The volume (VVM), the surface density of mitochondria (SVMO) and their mean profile size (am) were measured at the electron microscopic level (X 15 000). The most striking differences were observed in the volume density (VVM), surface to volume ratio (SV) and mean profile area of the mitochondria (am). Qualitative results revealed that the zone of good electron microscopic observation varied according to the concentrations of the fixative. In conclusion, the best qualitative and quantitative preservation is assumed with 4% paraformaldehyde and at a less degree with 6% paraformaldehyde + 0,25% glutaraldehyde.

Animals↗

Ultrastructural localization of transferrin synthesis in rat hepatocytes during prenatal and postnatal development.

The synthesis of transferrin has been investigated in rat hepatocytes during pre- and postnatal development of the liver by ultrastructural immunoperoxidase cytochemistry. Transferrin was detected in fetal and postnatal hepatocytes on the rough endoplasmic reticulum and in the Golgi apparatus, but the amount and distribution of the protein varied according to the stage of the hepatocyte maturation. Between days 13 and 19 of gestation, nearly all hepatocytes exhibited transferrin. After day 20 of pregnancy more and more unstained hepatocytes were detected. Two periods (19th day of gestation and one day after birth) were characterized by an intense labeling of the Golgi apparatus; in contrast, only a small amount of transferrin was located in this organelle at other stages of the rat liver development. The accumulation of transferrin in the Golgi apparatus is interpreted as a transitory storage of the protein. Transferrin could be produced by immature as well as mature hepatocytes.

Animals↗

[Ultrastructural demonstration of transferrin synthesis in the adult rat liver].

An ultrastructural immunoenzymatic study allowed us to demonstrate in Rat adult hepatocytes the presence of transferrin on the rough endoplasmic reticulum and in the Golgi apparatus. Such localizations suggest that the different steps of transferrin synthesis could possibly be related topographically to them. These results further an approach to the problems of maturation and secretion of transferrin.

Animals↗

Demonstration of the simultaneous presence of transferrin, hemopexin and albumin in the same adult rat hepatocyte.

The location of three plasma proteins (transferrin, hemopexin, and albumin) in hepatocytes was investigated in adult rats. The synthesis of transferrin anf hemopexin has been established by ultrastructural studies showing a labeling of the rough endoplasmic reticulum (RER). By using an indirect immunoenzymatic method with monospecific antibody solutions, the three proteins were detected in the same hepatocyte. The simultaneous presence of different plasma proteins in hepatocytes seems to point to the fact that the synthesis in these cells could be a non-specialized type.

Animals↗

Lack of short-latency-potentials in the VEP reflects immature extra geniculate visual function in delayed visual maturation (DVM).

PURPOSE: To investigate children with delayed visual maturation (DVM) and correlate the electrophysiological findings to visual development. METHODS: Three children, one from each of the DVM-classification groups, were subjected to routine ophthalmological examinations and electrophysiological examinations: flash visual evoked potential (VEP) and skin electrode electroretinography (ERG). RESULTS: All three children had normal ERGs but initially abnormal VEP-recordings with marked delay of latency or grossly altered VEPs. When visual interest developed with responsive smiling at 4, 4.5 and approximately 12 months of age, a maturation in the VEPs also appeared, with development of a short-latency complex (approximately 70 ms). In the normal neonatal development of the VEP, a negativity at approximately 60-70 ms (N1) emerged at four to six weeks of postnatal life when the child started responsive smiling and showing raised visual interest. According to animal experimental research and human studies, the development of the specific response (the short-latency complex) represents the gradual onset of cortical activity mediated via the specific retino-geniculo-striatal pathway. Thus, when the short-latency complex of the VEP cannot be identified, the visual function is mainly of subcortical origin. Since the VEP developed in the same way in the children with DVM as in normal subjects, the pathophysiological dysfunction and origins of DVM can partly be understood. CONCLUSIONS: The results show that i) children with DVM has a period of visual inattentiveness at a time when normal children show visual interest, ii) the VEP is abnormal in children with DVM at the time of visual inattentiveness, iii) the improvement of vision in DVM can be measured with VEP and iiii) the extra-geniculate system(s) provides for the visual function early neonatally in the normal child and in a prolonged period in the DVM-child as long as the VEPs are abnormal.

Electroretinography↗

The dual nature of lateral eye movements.

This study investigated the validity of lateral eye movements (LEM) as a measure of the individual differences and task demands aspects of LEM by comparing eye movements to information questions that had no clear hemispheric locus with eye movements to rhyme questions that demanded specific left hemisphere involvement. Fifty subjects were asked to respond to the two types of questions and the initial direction of their eye movement was recorded. The results indicated that LEM are a reliable measure of individual differences because both test-retest and split half reliabilities were high (.78 to .85). LEM were also shown to be responsive to task demands. The number of leftward LEM did not differ from rhyme and information questions, but there were more rightward LEM and fewer stares to the rhyme question. These results suggest that LEM are a reliable individual difference measure and are sensitive to task differences. There is clear evidence that LEM have neuropsychological implications, but every task demands the integrated functioning of the whole brain and these implications should not be overgeneralized.

Adolescent↗