Showing papers by "Uppsala University published in 1985"

Molecular cloning of Ancient Egyptian mummy DNA

Abstract: Artificial mummification was practised in Egypt from ∼ 2600 BC until the fourth century AD. Because of the dry Egyptian climate, however, there are also many natural mummies preserved from earlier as well as later times. To elucidate whether this unique source of ancient human remains can be used for molecular genetic analyses, 23 mummies were investigated for DNA content. One 2,400-yr-old mummy of a child was found to contain DNA that could be molecularly cloned in a plasmid vector. I report here that one such clone contains two members of the Alu family of human repetitive DNA sequences, as detected by DNA hybridizations and nucleotide sequencing. These analyses show that substantial pieces of mummy DNA (3.4 kilobases) can be cloned and that the DNA fragments seem to contain little or no modifications introduced postmortem.

Preparation of Pure Hepatocytes and Reticuloendothelial Cells in High Yield From a Single Rat Liver by Means of Percoll Centrifugation and Selective Adherence

Abstract: A rapid method for mass isolation of functionally intact hepatocytes and reticuloendothelial cells from a single rat liver is described. The technique is based on collagenase perfusion of the liver, isopycnic sedimentation in Percoll, and selective adherence of the cells. The Kupffer cells (KC) attach and spread on glass or plastic in serum-free medium 15 min following seeding. Cultures of KC are 90%-95% pure with about 5% liver endothelial cells (LEC), less than 1% parenchymal cells (PC) and a maximum of 5% stellate cells (SC). The LEC adhere and spread on fibronectin 60-120 min following seeding, forming cultures that are contaminated with 5-10% SC and less than 1% KC and PC. The yield of plated LEC is 50-60 X 10(6) per 200-g rat. Ultrastructural analysis shows that Percoll does not associate with the cells during the separation procedure.

The temporal evolution of hypoglycemic brain damage. I. Light- and electron-microscopic findings in the rat cerebral cortex.

Abstract: In the course of a study on the pathogenesis of neuronal necrosis in severe hypoglycemia, the morphological characteristics reflecting reversible and irreversible neuronal lesions were examined as a function of time following normalization of blood glucose. To that end, closely spaced time intervals were studied in the rat cerebral cortex before, during, and up to 1 year after standardized pure hypoglycemic insults of 30 and 60 min of cerebral isoelectricity.

T lymphocytes in collagen II-induced arthritis in mice. Characterization of arthritogenic collagen II-specific T-cell lines and clones.

Abstract: Collagen type II-spedfic long-term cultured T helper cells, derived from the DBA/1 mouse, have been established and characterized. Clones from these T-cell lines could he shown to recognize either species-specific or species-nonspecific determinants on the collagen type II molecule, including determinants on autologous mouse collagen. Induction of arthritis via transfer to both irradiated and normal syngeneic recipient mice was obtained with both collagen type II-specific T-cell lines and an autoreactive and collagen type II-specific T-cell clone. Fewer cells were needed to evoke arthritis in normal than in irradiated recipients. Cells from lines and the clone used for transfer were by immunocytochemistry shown to have T helper phenotype.

Concentration of sodium hyaluronate in serum.

Abstract: A radioassay for sodium hyaluronate using high-affinity binding protein from bovine cartilage has been modified for serum analysis. The accuracy of the method was checked by isotope dilution experiments and by recovery studies with exogenous hyaluronate. The between-assay standard deviation in the determination is 15-20%. The concentration of sodium hyaluronate in healthy adults (blood donors) is in the range of 10 to 100 micrograms/l with a mean value in the order of 30 to 40 micrograms/l. This is a lower concentration than previously reported. The same level was found in young people. Higher hyaluronate concentrations were noted in persons above 50 years of age. Analysis of plasma showed a slightly higher average hyaluronate level (5%) than in serum from the same persons. There were no notable sex differences. Analysis of serum and plasma from adult animals (rat, rabbit, dog, pig, goat, sheep, cow and horse) gave hyaluronate concentrations of the same order or higher than in human serum.

Antibodies against platelet-derived growth factor inhibit acute transformation by simian sarcoma virus

Abstract: A clue to the molecular mechanism of neoplastic transformation was provided by the finding of a near identity in amino-acid sequence between the platelet-derived growth factor (PDGF) B-chain and a region in the transforming protein, p28sis, of simian sarcoma virus (SSV), an agent that causes sarcomas and gliomas in experimental animals. This finding infers a direct link between the molecular biology of normal mitogenesis and oncogenesis since it suggests that the transforming activity of SSV is caused by a growth factor. Although PDGF agonist activity has been isolated from conditioned medium of SSV-transformed cells, it is not clear whether infection of responsive cells by SSV leads solely to autocrine stimulation of growth by a secreted PDGF-like factor or whether other, possibly intracellular, activities of p28sis or its processed products contribute to the transformation. To distinguish between these possibilities, we have studied the effect of anti-PDGF antibodies on acute SSV-transformation, and report here that these antibodies inhibit both proliferation and SSV-induced morphological changes in human diploid fibroblasts.

The chemistry and biochemistry of C-nucleosides and C-arylglycosides.

Abstract: Publisher Summary This chapter reviews recent advances in the chemistry and biochemistry of C-nucleosides, the literature concerning C-arylglycoside (i.e., nonnitrogen heterocyclic C-nucleoside) antibiotics, and recent significant advances in the synthesis of C-nucleosides and C-glycosides. It also discusses biological test data and data that are relevant to structure–activity relationships. Modification of readily available natural C-nucleosides is an attractive route to new C-nucleoside analogs and derivatives, because one starting material often possesses much of the desired functionality and chiral properties. The chapter illustrates this approach with examples. The most frequently used strategy for C-nucleoside synthesis involves the construction of a heterocyclic aglycone from the C-1 substituent of a functionalized sugar intermediate.

Platelet fatty acid composition in relation to fatty acid composition in plasma and to serum lipoprotein lipids in healthy subjects with special reference to the linoleic acid pathway.

Abstract: 1. The fatty acid composition in platelet phospholipids and in the plasma lipid esters as well as the serum lipoprotein lipid concentrations were determined in 67 healthy male subjects in order to establish the relationships between blood lipids and platelets. 2. A positive correlation was found between the concentrations of the triglyceride rich serum lipoprotein lipids and the relative percentage of saturated and monounsaturated fatty acids in plasma. The correlations were also positive between the serum high density lipoprotein-cholesterol concentration and the relative content of linoleic acid in the plasma cholesterol esters and phospholipids. 3. Negative correlations were found between the relative percentage of saturated and monounsaturated fatty acids in the plasma lipid esters versus linoleic acid in plasma and in the platelets. On the other hand there were positive correlations between linoleic acid in the plasma lipid esters and in the platelet phospholipids. These results indicate a direct dietary influence on the platelet phospholipid fatty acid composition. 4. The correlations between the fatty acids of the n −6 series within plasma and platelets as well as between plasma and platelets indicate that a high linoleic acid content is not associated with an increased arachidonic acid concentration. The results also indicate that the limiting metabolic step in the conversion of linoleic acid into arachidonic acid may be located at different levels in plasma and in the platelets.

DERMAL EFFECTS OF COMPOSITIONS BASED ON THE EUTECTIC MIXTURE OF LIGNOCAINE AND PRILOCAINE (EMLA) Studies in Volunteers

Abstract: The effects of the cutaneous application of EMLA cream (a eutectic mixture of lignocaine and prilocaine in their base form) were studied in volunteers. When tested by pin-prick, EMLA cream 2.5% and 5% produced analgesia of the area tested, the cream being most effective if left in contact with the skin for 60 min. The pain produced by the insertion of an i. v. cannula was successfully blocked by the application of this formulation, especially if applied to the antecubital area. Temporary blanching of the skin areas was frequently observed on removal of the occlusive tape bandages, but prolonged, or repeated, application of 5% EMLA cream did not produce local skin reactions. Tests for delayed hypersensitivity reactions were negative. Plasma concentrations of lignocaine and prilocaine were low after a standard application.

Absolute cross sections for multi-electron processes in low energy Arq+−Ar collisions: Comparison with theory

Abstract: Absolute cross sections have been measured for a variety of multi-electron processes in low-energy collisions of multiply charged argon recoil ions with neutral argon. The cross sections are compared with theoretical estimates based on an extension of the classical barrier model. Comparison is also made with the statistical theory of Muller et al.

Uptake of circulating hyaluronic acid by the rat liver. Cellular localization in situ.

Abstract: The uptake of [3H]acetyl-labelled hyaluronic acid (HA) intravenous was examined in the liver, spleen and kidney of the rat after i.v. injection. 3H-activity was located by light- and electron-microscopic autoradiography after measurement by scintillation counting of tissue digests. In the liver, approximately 90% of the radioactivity was located in the sinusoidal endothelial cells, with autoradiographic grains distributed throughout the cytoplasm; 50% of the grains overlay vacuoles 0.3 to 1.2 μm in diameter. A few grains (4%) were located in Disse's space or nearby in the cytoplasm of hepatocytes. No grains were found in Kupffer cells. The remainder were randomly scattered across the sections in a pattern indicating nonspecific background activity. These observations are in accordance with the selective uptake of HA exhibited by dissociated liver cells in vitro. HA concentrations in the spleen and kidney were too low for detection by autoradiography. Splenic concentrations were much lower than in rabbits or mice; in this respect the uptake of circulating HA in the rat resembles that reported for chondroitin 4-sulphate.

Corticotropin-Releasing Factor-like Immunoreactivity in Senile Dementia of the Alzheimer Type: Reduced Cortical and Striatal Concentrations

Abstract: The concentration of corticotropin-releasing factor-like immunoreactivity (CRF-LI) in the human central nervous system was measured by radioimmunoassay in postmortem tissue of control patients and in those with histologically confirmed senile dementia of the Alzheimer type (SDAT) In the controls, CRF-LI was found in high concentrations in the hypothalamus and frontal cortex (Brodmann's area 10), in moderate concentrations in amygdala, substantia innominata, temporal and parietal cortex (Brodmann's areas 38 and 7), and the caudate nucleus, and in low concentrations in posterior hippocampus and nucleus accumbens A marked reduction in the concentration of CRF-LI was observed in the frontal and temporal cortex (approximately 50%) as well as in the caudate nucleus (approximately 70%) in the SDAT group The present findings suggest that neurons containing corticotropinreleasing factor are pathologically altered in SDAT, in addition to the previously described cholinergic and somatostatinergic neuronal degeneration ( JAMA 1985;254:3067-3069)

Exocytosis of the prophenoloxidase activating system from crayfish haemocytes

Abstract: Lipopolysaccharides (LPS) and the β-1,3-glucan laminarin G, both of which specifically activate the prophenoloxidase (proPO) activating system of crayfish haemocyte lysate, were found to induce degranulation (exocytosis) and subsequent lysis in vitro of monolayers of semigranular haemocytes from the crayfish,Pacifastacus leniusculus, (Table 1, Fig. 1 b), whereas the granular cells were unaffected (Fig. 1 c). Exocytosis of isolated semigranular or granular cells in vitro could also be evoked by the Ca2α ionophore A23187 (Table 2, Fig. 1 d). In this case, the whole proPO system was released from the cellular vesicles in its inactive form, since the secreted material contained protease and prophenoloxidase as inactive proenzymes, which could be activated if LPS or β-1,3-glucans were added (Table 3). The anion channel blocker SITS, which inhibits exocytosis in several systems, prevented degranulation triggered by β-1,3-glucan, LPS, or ionophore. It is concluded that, in arthropods, LPS serve as an indicator of Gram negative bacteria and β-1,3-glucan as an indicator of fungi. These non-self molecules elicit both the exocytotic release of the proPO system from the semigranular cells and the subsequent biochemical activation of this system.

Functional and morphological characterization of cultures of Kupffer cells and liver endothelial cells prepared by means of density separation in Percoll, and selective substrate adherence.

Abstract: This paper presents a study on the structure and function of Kupffer cells (KC) and liver endothelial cells (LEC) isolated by a simple and rapid technique involving 1) perfusion of the liver with collagenase; 2) cell separation by means of density centrifugation in Percoll; and 3) cell culture, taking advantage of the fact that KC and LEC differ in their preferences for growth substrate. The KC, which attach and spread under serum-free conditions on surfaces of glass or plastic during the first 15 min in culture exhibit a typical macrophage-like morphology including membrane ruffling and a heterogenous content of vacuoles. Moreover, these cells express (a) Fc receptors (FcR) for binding and phagocytosis of erythrocytes covered with immune globulin G (E-IgG), and (b) complement receptors (CR) for binding and serum dependent phagocytosis of erythrocytes covered with either human C3b or mouse inactivated C3b (iC3b). The cells also bind fluid phase fluoresceinated C3b. Approximately 30% of the KC express immune response-associated (Ia)-antigens.

Synchronous short-term population fluctuations of some birds and mammals in Fennoscandia - occurrence and distribution

Abstract: We examined population fluctuations of the voles Microtus agrestis L. and Clethrionomys glareolus Schreb. by biannual trapping; black grouse Tetrao tetrix L., mountain hare Lepus timidus L., and red fox Vulpes vulpes L. by using bag records from all over Sweden, questionnaires from south-central Sweden, and population indices from Grims6 Wildlife Research Area in south-central Sweden. Synchronous population fluctuations between voles as a group and the other species conformed to a 3-4 year periodic pattern in both autumn and spring populations only in northern and central Sweden (the boreal forest region). Spring populations of boreal forest grouse, hare, and probably also fox, lagged one year behind the voles. The northerly areas also formed geographical units of co-fluctuation within each game species. Using our own data as well as reviewing previous studies in Fennoscandia, we conclude that synchronized 3-4 year population fluctuations of voles and medium-sized herbivores are confined to the central and northern part of Fennoscandia, although voles may exhibit short-term population fluctuations further south. The synchronizing link between the herbivores could be (1) food plant quality and/or quantity and/or (2) predation. We could not reject either of these two plausible mechanisms as a cause of interspecifically synchronous fluctuations.

The development of antioxidant enzymatic defense in the perinatal rat lung: activities of superoxide dismutase, glutathione peroxidase, and catalase.

Abstract: This report describes the development of superoxide dismutase, glutathione peroxidase, and catalase activities in fetal (days 16–22) and neonatal (day 2 postpartum) lungs of normal rats. Each of the enzymes showed an individual pattern of development in the perinatal period. Glutathione peroxidase activity increased by 135% (p < 0.001) during the last 3 days before birth, catalase activity by 105% (p < 0.01) during the first 2 postnatal days, and the activity of superoxide dismutase by 52% (p < 0.05) from gestational day 19 to 2 days after birth. Contamination by superoxide dismutase from blood in the lung samples accounted for less than 2% of the activity. In contrast, not less than 10–30% of glutathione peroxidase and catalase originated from the blood; and corrections were made for this source of error in each sample.

Differential effects of reserpine and 6-hydroxydopamine on neuropeptide Y (NPY) and noradrenaline in peripheral neurons.

Abstract: 1. The effects of 6-hydroxydopamine (6-OHDA) and reserpine pretreatment on peripheral neuropeptide Y (NPY)-and noradrenaline (NA)-containing neurons were studied in guinea-pigs. 2. Ten days after 6-OHDA pretreatment, a 60–80% reduction of the NA content was observed in the right atrium of the heart, stellate ganglion and spleen. The content of NPY-like immunoreactivity (LI) was reduced by about 50% in the heart, not changed in the spleen while it increased to 200% of control in the stellate ganglion. Immunohistochemistry showed a pronounced loss of NPY-and tyrosinehydroxylase (TH)-immunoreactive (IR) nerves in the heart but not in the spleen. Increased NPY-IR was seen in axons and cell bodies of the stellate ganglion. 3. Reserpine pretreatment (thereshold dose 0.5 mg x kg−1) caused a dose-and time-dependent reduction of the content of NPY-LI in the heart. A maximal depletion of NPY-LI (about 80%) was observed 5 days after reserpine. Reserpine pretreatment also reduced the content of NPY-LI in the spleen, while no significant change was observed in the adrenal gland or vas deferens. The levels of NPY-LI increased in the stellate ganglion to about 180% of control 5 days after reserpine. Immunohistochemical analysis revealed an almost total loss of NPY-IR nerve fibres in the heart as well as around blood vessels in the lung and skeletal muscle. No detectable changes were observed in perivascular NPY-IR nerves in the spleen, vas deferens or kidney. TH-IR nerves remained unchanged after reserpine, thus indicating that the observed loss of NPY-IR nerves was due to a depletion of NPY and not a degeneration. 4. No change in the levels of substance P-LI was observed in the right atrium 5 days after reserpine. 5. NA was, in contrast to NPY, markedly depleted in all tissues investigated after reserpine treatment. The depletion of NA was more extensive, and occurred more rapidly and at much lower doses as compared to the effects on NPY-LI. 6. Ligations of the sciatic nerve revealed that NPY-LI 4 was transported axonally with a rapid rate (3 mm/h). Reserpine pretreatment significantly increased the amount of accumulated NPY-IR above the ligation, suggesting an increase in axonal transport. 7. High performance liquid chromatography revealed that the NPY-LI consisted of two major peaks in the stellate ganglia, while only one peak closely corresponding to porcine NPY was seen in the right atrium. 8. In conclusion, 6-OHDA pretreatment depletes NPY-LI in certain terminal regions and increases NPY-LI in ganglia. Reserpine induces a tissue-and dose-dependent depletion of NPY-LI in certain terminal areas, while corresponding cell body content and axonal transport of the peptide seem to increase.