Fucosidosis and blood group substances in the urine]

Glycopeptide and oligosaccharide fractions obtained from fucosidosis urine contains more Lea activity (4-8 fold) than control urine. Both fucosidosis fractions also contained Leb and H activities, no A activity in contrast to the salivary and erythrocyte phenotypes (A, Le a+ b-). The amount of Leb activity is lower in both fractions (1) and (2) than that of Leb control children (4-16 fold decrease). Blood group A activity was not detected at any concentration used (less than or equal to 50 fold-concentrated urine) whereas A activity was found in (A, Le a- b+) control urine. On the contrary, both fucosidosis fractions contained H activity, whereas A (Le a- b+) control children fractions had none (less than or equal to 50-fold concentrated urine). H and Leb activity might originate from Lea precursor apart from the "non-secretory" type of the patient.     

Distribution of CSF (colony stimulating factor) in kidney of the mouse.

The activity of the colony stimulating factor (SCF) was measured in kidney subcellular fractions in mice. The highest activity was noted in microsomes. From other fractions, the cytosol had large amounts of CSF. On the basis of literature data, and the findings presented we suggest that the kidney is at least one of the organs of CSF biosynthesis.     

Distribution of CSF (colony stimulating factor) in kidney of the mouse

Summary The activity of the colony stimulating factor (CSF) was measured in kidney subcellular fractions in mice. The highest activity was noted in microsomes. From other fractions, the cytosol had large amounts of CSF. On the basis of literature data, and the findings presented we suggest that the kidney is at least one of the organs of CSF biosynthesis.     

Fractionnement des protéines bactériennes par électrophorèse en gel d'acrylamide-agarose

Summary Acrylamide-agarose gel electrophoresis was employed to separate proteins obtained by sonication of various gram negative bacilli. The bulk of bacterial protein migratc at pH 8.7 to the anode, but often some fractions are quickly migrating to the cathode. Some 30 fractions can be distinguished this way. This technique proves interesting for the study of microbial protein metabolism and may easily be applied to numerous bacteria.     

Effect of exercise on ribonuclease activity in rat skeletal muscle.

Distribution of ribonuclease activity (measured at pH 7.6) in subcellular fractions of homogenates of rat skeletal muscle was investigated in sedentary animals and after 8 weeks running program. Training increased ribonuclease activity (expressed as units of enzyme per g of muscle protein). There was no increase in nuclear fraction, but in both cytoplasmic and mitochondrial fractions the RNA-ase activity increased 42% and 45% respectively.     

Use of a zonal rotor for demonstrating that human myelin is formed by a continuum of particles of different densities]

Human myelin isolated from frozen brain consists of a continuum of particles as shown by sucrose continuous gradient (between 0,4 and 0,85 M) on zonal rotor. This was determined by measuring the O. D. at 260 and 280 millimicron, the protein concentration by Lowry method and sucrose concentration by polarimetry in 60 fractions. Most of the material is concentrated between 20-23% sucrose, the maximum being found at 21.5% (0.63 M). The fractions are dialyzed for 62 hrs. to remove the sucrose and the pellet obtained after centrifugation shows that the protein concentration increases from the ligher fraction to the heavier.     

In vitro protein synthesis and alpha amylase activity in F cells from hepatopancreas of Palaemon serratus (Crustacea; Decapoda).

In crustaceans, all the steps in the assimilation of food take place in the hepatopancreas. To facilitate the study of this organ, a method for the dissociation of cell types was developed. The hepatopancreas of the prawn Palaemon serratus was mechanically dissociated and the cells separated by Percoll density-gradient centrifugation. The E and R cells had similar densities of around 1.05 g/ml. The F cells were separated into two distinct fractions with densities of 1.075 and 1.082 g/ml. The B cells sedimented at a density of 1.12 g/ml. The ratio between the two populations of F cells was found to vary during the intermolt cycle while B cells disappeared after the molt. When the density gradient fractions were incubated with 3H-leucine, incorporation was highest in the F cell fractions. Measurements of alpha-amylase activity, indicated that the two populations of F cells may be derived from the same cell type.     

Adenosine 5'-triphosphate sulphurylase from Spirulina platensis.

ATP-sulphurylase from an unicellular blue-green alga, Spirulina platensis was localized in the soluble fractions of cell-free homogenate, and it was stable for over 3 weeks at -6 degrees C.     

Use of 3H-QNB in the isolation of plasma membrane from smooth muscle of the urinary bladder: effect of oxalate on calcium uptake by the membrane fractions

Specific binding of tritiated quinuclidinyl benzilate (3H-QNB) to surface membrane muscarinic receptors was utilized to identify plasma membrane (PM) fractions from smooth muscle of the rabbit urinary bladder. Accumulation of 3H-QNB in the PM fraction was 4-5-fold higher than that in fractions of endoplasmic reticulum (EM) or mitochondria (M). A similar pattern of distribution was found for 5'-nucleotidase. 3H-QNB binding therefore appears to be a suitable marker for plasma membrane of the urinary bladder. Data on ATP-dependent calcium uptake by PM and ER fractions showed that oxalate highly potentiated calcium uptake by both fractions and consequently this feature cannot be used to identify ER fractions specifically.     

Leukozytenemigrationsfördernde Wirkung von Crotonöl

Summary Various croton oil fractions were studied with regard to theirin vitro effect on leucocytes and their irritative action. It was discovered that there exist fractions exerting a stimulating effect on leucytic migration and an irritative action and other fractions having a leucocytic stimulating effect without pronounced irritative action. The co-carcinogenic fractions ofDanneel have no promotory effect on the migration of leucocytes.     

Effect of exercise on ribonuclease activity in rat skeletal muscle

Summary Distribution of ribonuclease activity (measured at pH 7.6) in subcellular fractions of homogenates of rat skeletal muscle was investigated in sedentary animals and after 8 weeks running program. Training increased ribonuclease activity (expressed as units of enzyme per g of muscle protein). There was no increase in nuclear fraction, but in both cytoplasmic and mitochondrial fractions the RNA-ase activity increased 42% and 45% respectively.Acknowledgments. I would like to thank Prof. L. elewski and Dr J. Popinigis for helpful suggestions.     

Changes in transglutaminase activity in carbon tetrachloride-damaged rat liver

Summary A significant decrease in transglutaminase (TGase) activity was observed in the cytosol and nuclear fractions of carbon tetrachloride-damaged rat liver. The degree of decrease in TGase activity in the cytosol fraction was closely related to the serum transaminase level. Gel filtration studies revealed that TGase activity in 80 kDa fractions significantly decreased, but that in 160 kDa fractions slightly increased after carbon tetrachloride treatment.     

Changes in transglutaminase activity in carbon tetrachloride-damaged rat liver.

A significant decrease in transglutaminase (TGase) activity was observed in the cytosol and nuclear fractions of carbon tetrachloride-damaged rat liver. The degree of decrease in TGase activity in the cytosol fraction was closely related to the serum transaminase level. Gel filtration studies revealed that TGase activity in 80 kDa fractions significantly decreased, but that in 160 kDa fractions slightly increased after carbon tetrachloride treatment.     

Adenosine 5′-triphosphate sulphurylase fromSpirulina platensis

Summary ATP-sulphurylase from an unicellular blue-green alga,Spirulina platensis was localized in the soluble fractions of cell-free homogenate, and it was stable for over 3 weeks at –6°C.     

Protective effect of Shigyaku-to,a traditional Chinese herbal medicine,on the infection of herpes simplex virus type 1 (HSV-1) in mice

The antiviral activity of Shigyaku-to (TJS-109), a traditional Chinese herbal medicine, was investigated in mice infected with herpes simplex virus type 1 (HSV-1). TJS-109 is a combination of the medicinal plant extracts fromZingiberis siccatum rhizoma,Aconiti tuber andGlycyrrhizae radix in a specific proportion. Mice infected with a 10 LD₅₀ dose of HSV-1 were treated with TJS-109 orally at doses of 1.25 to 20 mg/kg 2 days before, and 1 and 4 days after the infection. The treated groups had 80% (1.25 mg/kg), 40% (5 mg/kg) and 23% (20 mg/kg) mortality rates 25 days after the infection as compared with a 100% mortality rate in control mice treated with saline. When HSV-1 infected mice (recipients) received CD8⁺T cell fractions derived from spleens of mice treated with TJS-109 (donors), 70% of recipients survived, as compared with 0% survivors in the groups of mice treated with saline, B cell fractions, CD4⁺ T cell fractions or macrophage-enriched fractions prepared from the same donors. TJS-109 did not show any virucidal activities against HSV-1 or any virostatic activities on the growth of HSV-1 in Vero cells. These results suggest that TJS-109 protected mice exposed to lethal amounts of HSV-1 through the activation of CD8⁺ T cells.     

Ribonucleaseaktivität isolierter Nebennierenmarkgranula

Summary Homogenates from suprarenal medulla of cattle were fractionated by density gradient centrifugation. The catecholamine and RNA content of all fractions obtained, as well as their RN-ase activity, were determined and expressed as % per mg protein. The chromaffin granules contain 49.8% of the catecholamines, 15.7% of the RNA, and 18.7% of the RN-ase activity of all fractions. The demonstration of RN-ase activity in the chromaffin granules is in favour of the hypothesis that this enzyme could catalyse the spontaneous release of RNA, catecholamines, and ATP at 37°C.

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Ausgeführt mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Functional and biochemical characteristics of mitochondrial fractions from rat liver in cold-induced oxidative stress

We determined characteristics of rat liver mitochondrial fractions, resolved at 1000 (M₁), 3000 (M₃), and 10,000 g (M₁₀) after 2 and 10 days cold exposure. In all groups, the M₁ fraction exhibited the highest oxidative capacity, oxidative damage, H₂O₂ production rate, and susceptibility to stress conditions, and the lowest antioxidant levels. Cold exposure increased cytochrome oxidase activity in all fractions and succinate-supported O₂ consumption in the M₁ and M₁₀ fractions during state 3 and state 4 respiration, respectively. With succinate, the H₂O₂ release rate increased in all fractions during state 4 and state 3 respiration, whereas with pyruvate/malate, it increased only during state 4 respiration. Increases in tissue mitochondrial proteins caused a faster H₂O₂ flow from the mitochondrial to cytosolic compartment, which was limited by the reduction in the M₁ fraction. Despite increased liposoluble antioxidant levels, cold also caused enhanced oxidative damage and susceptibility to oxidative challenge and Ca²⁺-induced swelling in all fractions. These changes leading to elimination of H₂O₂-overproducing mitochondria avoided excessive tissue damage. We propose that triiodothyronine, whose levels increase in the cold environment, brings about the biochemical changes producing oxidative damage and those limiting its extent.Received 16 July 2004; received after revision 27 September 2004; accepted 18 October 2004     

In vitro protein synthesis and α amylase activity in F cells from hepatopancreas ofPalaemon serratus (Crustacea; Decapoda)

In crustaceans, all the steps in the assimilation of food take place in the hepatopancreas. To facilitate the study of this organ, a method for the dissociation of cell types was developed. The hepatopancreas of the prawnPalaemon serratus was mechanically dissociated and the cells separated by Percoll density-gradient centrifugation. The E and R cells had similar densities of around 1.05 g/ml. The F cells were separated into two distinct fractions with densities of 1.075 and 1.082 g/ml. The B cells sedimented at a density of 1.12 g/ml. The ratio between the two populations of F cells was found to vary during the intermolt cycle while B cells disappeared after the molt. When the density gradient fractions were incubated with³H-leucine, incorporation was highest in the F cell fractions. Measurements of -amylase activity, indicated that the two populations of F cells may be derived from the same cell type.     

Effect of the extracts from Aristolochia indica Linn. on interception in female mice.

The crude petroleum ether, chloroform and alcoholic extracts from the roots of Aristolochia indica (Linn.) showed 100% interceptive activity in mature female mice at the single dose of 100 mg/kg body wt. The follow-up studies with the chloroform extract showed the most significant effect in the basic part and two acidic fractions at the single dose levels of 50 mg/kg body wt. No toxic effect was observed at the dose levels used.     

On the solvent-extraction of acetylcholine receptor

Summary Chloroform/methanol rechromatography on Sephadex LH-20 of fractions isolated from organic solvent extracts ofE. electricus electric organs exhibited alterations in elution point compared to the original chromatography. Additionally,³H decamethonium bromide did not coelute with any of the observed fractions. It is concluded that as presently described the organic solvent isolation procedure for the cholinergic receptor exhibits undesirable characteristics.Acknowledgments. The authors thank Dan Burrough and Rick O'Brien for their competent technical assistance. Naval Medical Research Development and Command, Research Work Unit No. MRO41.06.01.0021. The opinions and assertions contained herein are the private ones of the writers and are not to be construed as official or reflecting the views of the Navy Department or the naval service at large.