bluuebunny/crossref_metadata_2025 huggingface dataset:
Somatostatin binding sites in cytosolic fractions of parietal and non-parietal cells from rabbit fundic mucosa
Authors: Arilla Eduardo , Pilar Löpez-Ruiz M. , Gonzalez-Guijarro Luis , Prieto Juan C.
Published: February 2005

Specific binding sites for somatostatin have been found in the cytosolic fractions of both parietal and non-parietal cells from rabbit gastric fundic mucosa. The stoichiometric data suggested the presence of two classes of binding sites in both types of ceils. The number of low-affinity binding sites was significantly higher in parietal cells than in non-parietal cells. The reverse was true for the high-affinity binding sites. However, the affinity of each class of binding sites was similar in the cytosolic fractions of both parietal and non-parietal ceils. It thus appears that low-affinity somatostatin binding sites are mainly located in the parietal ceils whereas the high-affinity sites occur principally in the non-parietal cells.
Functional prokaryotic gene control signals within a eukaryotic rainbow trout protamine promoter
Authors: Jankowski Jacek M. , Walczyk Eva , Dixon Gordon H.
Published: February 2005

Following the construction of a series of pSV2-cat derived plasmids containing the chloramphenicol acetyltransferase (CAT) gene under the control of a eukaryotic trout protamine promoter, it was noted thatEscherichia coli, transformed with these plasmids, developed resistance to chloramphenicol (CM). This result suggested that the eukaryotic trout protamine promoter possessed significant prokaryotic promoter activity. Modification of the trout protamine promoter region by removing the region containing the eukaryotic Goldberg-Hogness box in the plasmid p525-cat increased the expression of the CAT gene almost to the wild-type level and conferred strong CM resistance. Sequence comparisons of the plasmid series indicate that prokaryotic promoter elements are present in the trout protamine promoter and that their similarity to the prokaryotic promoter consensus sequences and the distance between the two elements is more favourable in p525-cat, the plasmid which conlers the greatest CM resistance.
Bovine elastin cDNA clones: Evidence for the occurrence of a new elastin-related protein in fetal calf ligamentum nuchae
Authors: Raju K. , Rampersad V. , Pulleyblank D. E. , Krawetz S. A. , Anwar R. A.
Published: February 2005

Poly (A+) mRNA was isolated from fetal calf ligamenturn nuchae and used for the construction of cDNA libraries. A fraction highly enriched in elastin mRNA was used to prepare the cDNA probes for screening the libraries. A 2 kb clone, pREl, gave the most positive signal in colony hybridization. It hybridized to a mRNA of the same size as reported for elastin mRNAs from chick and sheep. Hybrid-arrested translation showed that translation of mRNAs for proteins other than elastin doublet was not inhibited by pREI. Southern blot analysis showed that pREl has sequence homology with pVE6 and pVE10, which were tentatively identified as elastin-related cDNA clones representing two distinct mRNAs. DNA sequence data from the 5′ end of pREl show that the translated amino acid sequence is not typical of known elastin sequences but contains some elastin-like sequences. All of this evidence strongly suggests the occurrence in fetal calf nuchal ligament of a mRNA which codes for a previously unknown elastin-related protein.
Regulation of tubulin by triiodothyronine in hypothyroid rat brain
Authors: Mazumder Anjana , Das Kamal , Sarkar Pranab K.
Published: February 2005

The effect of T3 (triiodothyronine) on the induction of tubulin in hypothyroid developing rat brain has been examined using organ cultures of brains from late fetal, neonatal and postnatalrats. The neonatal brain displayed maximum sensitivity to T3. Hypothyroidism resulted in a 26% decline in the level of tubulin in the neonatal brain as opposed to a 5–15% decline in the fetal or postnatal brain. Exposure of the hypothyroi d neonatal brain to T3 for 2 h in culture led to a 61% rise in the level of tubulin in contrast to a 41% increase seen in the case of normal brain. Total protein synthesis was not significantly affected. The preferential decline of tubulin in the neonatal hypothyroid brain, its enhanced sensitivity to T3 compared to normal brain, and the coincidence of the period of sensitivity to that of brain maturation indicate that the regulation of the level of tubulin by T3 in the developing brain is a natural ontogenic phenomenon.
The effect of osmotic pressure of aqueous PEG solutions on red blood cells
Authors: Herrmann Andreas , Arnold Klaus , Pratsch Lothar
Published: February 2005

A drastic increase of the intracellular microviscosity of red blood cells in the presence of polyethylene glycol (PEG) was established by electron spin resonance using the small spin label molecule 2,2,6,6-tetramethylpiperidine-N-oxyl-4-one (TEMPONE). The effective osmotic pressure of PEG solutions stressing the cells was estimated by comparison with those cytoplasmic rotational correlation times of TEMPONE measured in NaCl or sucrose containing media of known osmotic pressure.
Amino acid sequence homologies between the high-mobilitygroup proteins, HMG-T from trout testis and HMG-1 and-2 from calf thymus: is the poly-aspartic-glutamic acid polypeptide within the main chain?
Authors: Watson D. C. , Dixon G. H.
Published: February 2005

The amino acid sequence of the N-terminal two-thirds of a trout high-mobility-group protein, HMG-T, has been determined as a continuous sequence of 174 residues out of a total for the whole molecule of 260 residues. When this sequence was compared with published sequences of long cyanogen bromide-derived peptides from the analogous calf-thymus proteins, HMG-1 and −2 (Walker et al., 1979), there was strong homology, with 60–70% identity of corresponding amino acid residues in the three proteins, the majority in lengthy identical runs. However, a discrepancy in the position of a highly acidic run of aspartic and glutamic residues suggests this region may not lie within the main polypeptide chain but may represent a separate chain or possibly a branched structure,
Alkalinization of phosphorylase kinase-deficient muscle during tetanic contraction
Authors: Lutaya G. , Rahim Z. H. A. , Shuttlewood R. J. , Bashford C. L. , Griffiths J. R.
Published: February 2005

The intracellular pH of resting and stimulated muscle was monitored by two independent methods: measurement of pH iniodacetate-treated homogenates of freezeclamped tissue and the absorbance at 550–443 nm of intracellular neutral red dye in vivo. During tetanic stimulation, muscle of phosphorylase kinase-deficient mice shows a transient alkalinization whereas muscle in normal mice becomes more acid under similar conditions. The alkalinization appears to be caused by abnormally rapid AMP deamination associated with adaptation to phosphorylase kinase deficiency.
Voltage-dependent gating properties of the channel formed by E. coli hemolysin in planar lipid membranes
Authors: Menestrina Gianfranco , Ropele Monica
Published: February 2005

Escherichia coli hemolysin forms cation selective, ion-permeable channels of large conductance in planar phospholipid bilayer membranes. The pore formation mechanism is voltage dependent resembling that of some colicins and of diphtheria toxin: pores open when negative voltages are applied and close with positive potentials. The pH dependence of this gating process suggests that it is mediated by a negative fixed charge present in the lumen of the pore. A simple physical model of how the channel opens and closes in response to the applied voltage is given.
Mechanisms of attack and defence at the cell surface: The use of phospholipid bilayers as models for cell membrane
Authors: Pasternak C. A. , Bashford C. L. , Menestrina G.
Published: February 2005

Electrical conductivity across phospholipid bilayers induced by various cytotoxic proteins has been used to analyse the damaging action of such proteins on cells; the protective effect of divalent cations and protons against such attack has also been investigated. The predominant effect of divalent cations and protons is to promote the closed state of membrane pores, i.e. to “gate” protein-induced lesions.
The in vitro transcription of a rainbow trout (Salmo gairdnerii) protamine gene. II. Controlled mutation of the cap site region
Authors: Jankowski Jacek M. , Dixon Gordon H.
Published: February 2005

A series of pJasmids containing new fusion genes in which the trout protamine gene is placed under the control of the complete herpes virus (HSV-I) tk promoter Pvu II-Bgl II fragment (pMg), or a shortened thymidine kinase (tk) promoter in which the region between the TATA box and the cap site is altered by using the Pvu II-Mlu I fragment (pM7), have been constructed. An additional recombinant plasmid was constructed in which the Bgl II-Ava II fragment of the protamine gene containing the entire protamine promoter but missing the protamine coding region was cloned into pBR322 between the Xho II 1666 and Hind III sites (pP5). For in vitro transcription, a HeLa cell lysate system was prepared and the RNA transcription products, after glyoxalation, were electrophoretically analyzed on 5% polyacrylamide gels. In constructing pM8 the DNA sequence between the tk promoter and the cap site was present while in pM7 it was deleted. Similar multiple transcripts were seen in both cases, indicating that the region between the promoter and the cap site has no effect upon transcription in vitro. The multiple transcripts appear to be due to the presence of a cryptic promoter in the complementary strand of the protamine gene. The activity of this cryptic promoter has been confirmed by comparison of the transcription of plasmid pPS, in which the protamine mRNA coding region has been deleted, with a previously described plasmid, p3BRP (Jankowski 3M and Dixon GH (1984) Can. J. Biochem. Cell. Biol. 62, 291–300), containing the intact protamine gene.
Partial sequence homologies between cytoskeletal proteins, c-myc, Rous sarcoma virus and adenovirus proteins, transducin, and β- and γ-crystallins
Authors: James M. , Crabbe C.
Published: February 2005

Computer based sequence comparisons indicate partial sequence homology between human c-myc, Rous sarcoma virus, adenovirus 7, and simian sarcoma virus proteins and the cytoskeletal proteins desmin, keratin and vimentin. In addition, sections of the oncogene proteins showed partial but significant homology to α and β subunits of transducin, γ-II and β-BP crystallins showed partial but significant homology to the cytoskeletal proteins keratin, vimentin, desmin, α and β-tubulin, and to adenovirus 7 and simian sarcoma virus transforming gene proteins. β-BP crystallin showed partial but significant homology to Rous sarcoma virus protein, and to α and y subunits of transducin. Both crystallins showed partial sequence homology to the GTP-binding protein elongation factor TU from Escherichia coli. These sequence homologies suggest a link between the mechanisms of normal lens cell differentiation, involving modifications to the cytoskeleton and subsequent changes to the pattern of protein synthesis, and mechanisms of neoplastic transformation. Furthermore the transducin-like region on β-crystallin may be important for its interaction with lens membranes and the maintenance of short-range order for lens transparency.
Crystallization of Ca2+ ATPase in sarcoplasmic reticulum vesicles by phospholipase treatment
Authors: Misra Manoj , Malhotra S. K.
Published: February 2005

Ca2+ ATPase molecules in sarcoplasmic reticulum, isolated from rabbit skeletal muscl% have been induced to crystallize into two-dimensional arrays by incubating the vesicles with phospholipase A2 and dialysing against dilute Tris/HCl buffer. These crystals differ in shape and size from those produced by treatment of the sarcoplasmic reticulum vesicles with Na3VO4. However, the unit-cell dimensions of both types of crystals are similar. The differences in shape and size are presumably due to differences in the mechanisms of crystal formation induced by treatment with phospholipase and Na3VO4.
Intermediate filament structure
Authors: Fraser R. D. B. , MacRae T. P.
Published: February 2005

In a previous communication (Biosci. Rep. 3, 517–525, 1993) we described quantitative X-ray diffraction studies of α-keratin which were shown to be consistent with the presence of finite arrays of repeating units, successive arrays being set down at axial intervals of 470 Å. In addition the axial interval between repeating units in an array was shown to be 197.9 Å. It was suggested that this could most readily be explained by supposing that a surfacelattice was present which contained a dislocation along a helical path with unit height h = 470 Å and unit twist |t| = 49.1°. The number of repeating units was shown to be in the range 7–9. With 7 repeats the mismatch of the lattice along the dislocation is small and this choice was used to develop a detailed model for the filament. Subsequent studies of molecular interactions have shown however that the coiled-coil rope segments in the rod domain of the molecule are most probably oriented parallel to the dislocation, and so minimization of lattice mismatch may be less important than originally supposed. In the present communication it is shown that the choice of 8, rather than 7, for the number of repeating units yields a model which is more compatible with estimates of the linear density and also provides the basis for a general model for polymorphism in intermediate filament lattices.
Sodium butyrate selectively inhibits host cell glycoprotein synthesis in human fibroblasts infected with cytomegalovirus
Authors: Radsak K. , Wiegandt H. , Unterdörfer G. , Wagner C. , Kaiser C. J.
Published: February 2005

Host cell as well as viral DNA synthesis in human fibroblasts infected with human cytomegalovirus was found to be largely resistant even to high concentrations sodium butyrate. Likewise, production of viral progeny was reduced by 1–2 orders of magnitude but not abolished. On the other hand, the drug allowed (modified) glycosylation only of viral polypeptides whereas that of host proteins was suppressed. Immunofluorescence studies on living cells suggested that butyrate may interfere with processing and intracellular transport of virus-specific surface membrane antigens.
Developmental study on the regulation of neurotransmitter-sensitive adenylate cyclase systems in primary cerebral cell cultures from embryonic mice
Authors: Shanker Gouri , Pieringer Ronald A.
Published: February 2005

An ontogenetic study of the effect of various neuhormones and other activators on adenylate cyclase systems was carried out using cultures of ceils from 15-d-old embryonic mouse brain. Dopamine stimulated the enzyme activity at earlier culture ages (i.e. 4 and 10 d) but had little stimulatory effect at later ages (i.e. 20 and 33 d). Further, this stimulation at the earlier ages was blocked by the dopaminergic blocker, fluphenazine, but not by α and β-adrenergic antagonists. In contrast to dopamine, isoproterenol (a β-adrenergic agonist) had little stimulatory effect at earlier ages, but its ability to stimulate cyclase activity increased with age. This increase in all age groups was blocked by propranolol (afg – adrenergic antagonists). Epinephrine-sensitive enzyme activity showed a steady increase with age, which could be blocked with propranolol except in β-d-old cultures, where it was blocked instead by fluphenazine. Because the cultures are relatively enriched in neurons at earlier ages and in glia in later ages, the results suggest a predominantly neuronal localization for the dopamine sensitive adenylate cyclases and a glial localization of the isoproterenol and epinephrine sensitive adenylate cyclases. Histamine, serotonin, calcium/calmodulin and chloroadenosine were either only slightly or not at all stimulatory.
Phospholipids as a possible instrument for translocation of nascent proteins across biological membranes
Authors: Rietveld Annie , de Kruijff Ben
Published: February 2005

The interaction of phospholipids with precursor proteins, particularly with the mitochondrial precursor protein apocytochrome c is reviewed and integrated with other aspects of protein insertion and translocation, leading to a model for (apo)cytochrome c import into mitochondria, in which phospholipids play a dominant role.
Solvation properties of ubiquinone-10 in solvents of different polarity
Authors: Ondarroa Monica , Sharma Santosh K. , Quinn Peter J.
Published: February 2005

The solvation properties of ubiquinone-10 and ubiquinol-10 in a wide variety of solvents of polarity varying from alkanes to water are reported. Greatest solubility is observed in solvents of intermediate polarity and particularly where low polarity is combined with a pronounced tendency to interact with the benzoquinone substituent of the ubiquinone molecule. This includes solvents like chloroform and benzene. Ubiquinone-10 is somewhat less polar than ubiquinol-10 as judged by comparative solubilities of the two molecules. Proton-NMR chemical shift measurements and aggregation studies in selected solvents indicate that in ubiquinone-10 in the liquid phase and in solution in hydrocarbons like dodecane the molecules have a preferred association possibly involving stacking of the benzoquinone rings. Surface balance studies indicated that the surface-active character of ubiquinone-10 is relatively weak and only in a comparatively polar and highly structured solvent, formamide, was there evidence of an effect on surface tension of the solvent. The critical micelle concentratiom in this solvent was estimated to be about 5 μM on the basis of surface tension measurements. Ubiquinone-10 is well known to form virtually insoluble monolayers at the air/water interface. Studies of the partition of ubiquinone-10 in binary mixtures of solvents suggest that the interaction of the benzoquinone ring substituent with structured polar solvents is considerably weaker than the internal cohesion between molecules of the solvent. No evidence on the basis of wide-angle X-ray diffraction measurements was obtained to indicate that solvent molecules were a component of the crystal lattice of ubiquinone-10 that had precipitated from solvent mixtures.
Rapid response of protein synthesis to insulin in 3T3 cells: Effects of protein kinase C depletion and differences from the response to serum repletion
Authors: Hesketh John E. , Campbell Gillian P. , Reeds Peter J.
Published: February 2005

Quiescent 3T3 cells grown in media containing 4% foetal calf serum showed different responses to insulin and to serum repletion (to 12%). Insulin stimulated protein synthesis within 1 h and this early response was insensitive to actinomycin D. The later insulin response showed progressive sensitivity to actinomycin D. The serum response was slower, not occurring until 1 h, and was inhibited by actinomycin D. Depletion of cell protein kinase C by pre-treatment with phorbol ester caused a total block of the immediate response to insulin but had little effect on the response to serum or the later response to insulin. Acute phorbol ester treatment stimulated protein synthesis.
Homologous IRCM-Serine Protease 1 from pituitary, heart atrium and ventricle: A common pro-hormone maturation enzyme?
Authors: Seidah Nabil G. , Cromlish James A. , Hamelin Josée , Thibault Gaétan , Chrétien Michel
Published: February 2005

IRCM-Serine Protease 1 (IRCM-SP1) has recently been isolated and characterized from porcine pituitary anterior and neurointermediate lobes (Cromlish et al., 1986a, J. Biol. Chem.261:10850–10858; Cromlish et al., 1986b, J. Biol. Chem.261:10859–10870). This pituitary serine protease was shown to selectively cleave human proopiomelanocortin (POMC)-derived peptides at both pairs of basic residues and C-terminal to specific Arg residues, all known to be cleaved in vivo. Here, a similar enzyme was isolated from rat heart atria and ventricles. Rat IRCM-SP1 was shown to be highly specific for the same cleavage sites in POMC, as the porcine pituitary homologue. Furthermore, the rat and the porcine enzymes cleave rat pro-Atrial Natriuretic Factor (pro-ANF 1–126) to yield ANF 103–126, 102–126 and 99–126 in that order of preference. This suggests that in vitro the cleavage sites preferred in pro-ANF resemble those found in brain and hypothalamus. The enzyme is nine times more abundant in atria versus ventricles/mg protein. It is concluded that IRCM-SP1, could well represent a common pro-hormone maturation enzyme for POMC and Pro-ANF and possibly many other pro-hormones.
The effect of islet amyloid polypeptide (amylin) and calcitonin gene-related peptide on glucose removal in the anaesthetized rat and on insulin secretion from rat pancreatic isletsin vitro
Authors: Tedstone Alison E. , Nezzer Tania , Hughes Stephen J. , Clark Anne , Matthews David R.
Published: February 2005

The effect of intravenous infusion of islet amyloid polypeptide (IAPP/amylin) and calcitonin gene-related peptide (CGRP) on blood glucose and plasma insulin in the basal and glucose-stimulated state was investigated in the anaesthetized rat. Both peptides had no effect on basal blood glucose or plasma insulin but following an intravenous bolus of glucose, CGRP-treated rats were hyperglycaemic and hyperinsulinaemic compared with control animals which were similar to IAPP-treated rats. IAPP had no effect on glucose-stimulated islet insulin secretion. These results suggest that CGRP, but not IAPP, alters glucose removal in vivo.
A role for G-proteins in the epidermal growth factor stimulation of phospholipase A2 in rat kidney mesangial cells
Authors: Hack Nashrudeen , Clayman Paula , Skorecki Karl
Published: February 2005

We have previously demonstrated phospholipase C (PLC) independent activation of phospholipase A2(PLA2) by epidermal growth factor (EGF) in glomerular mesangial cells in culture. In the current study using glass beads to permeabilize [3H]- or [14C]-arachidonate labelled mesangial cells we demonstrate that guanine nucleotides modulate the EGF-mediated stimulation of arachidonic acid release (75% inhibition with 100 μM GDPβS and 108% augmentation with 100 μM GTPγS). GTPγS alone stimulated both the release of free arachidonic acid and production of diacylglycerol (DAG), while EGF itself neither stimulated DAG nor augmented the DAG response to GTPγS. These findings suggest the intermediacy of a G-protein in PLC-independent stimulation of PLA2 by a growth factor, and provide a model system for determining the relationship between G-protein intermediacy and the intrinsic tyrosine kinase activity of the growth factor receptor.
Protein kinase C and casein kinase II activities in two human colon carcinoma cell lines, HT-29 and CaCo-2: Possible correlation with differentiation
Authors: Rydell Ewa , Magnusson Karl-Eric , Sjö Anita , Axelsson Krister
Published: February 2005

Protein kinase C (PK-C) and casein kinase II (CK-II) activities were studied in two human colon carcinoma cell lines (HT-29 and CaCO-2) undergoing differentiation in vitro resulting, in small-intestine-like cells. CaCo-2 cells, when grown under standard conditions, appear to undergo spontaneous differentiation. In these cells PK-C and CK-II activities were determined on day 5, 10 and 15. No significant differences in activities were seen either in PK-C or CK-II activity. HT-29 cells, when grown in glucose-free medium can be stimulated to undergo differentiation which is completed within 20 days. PK-C and CK-II activities were determined after 5, 10, 15, 20 and 25 days, respectively. PK-C activity rose from 7.9±3.5 pmole32P/mg protein/min at day 5 to 37.5±14.8 pmole32P/mg protein/min at day 20. After 25 days the activity was reduced to 20.0±7.8 pmole32P/mg protein/min. CK-II activity did not change significantly during day 5 to 20, but on day 25 there was a significant decrease in CK-II activity from 94.9±6.4 pmole32P/mg protein/min (day 20) to 62.6±3.9 pmole32P/mg protein/min (day 25) p=0.003. The results in this study indicate a role for PK-C and CK-II in cell growth and differentiation.
Periodic stimuli are more successful than randomly spaced ones for inducing development in Dictyostelium discoideum
Authors: Nanjundiah Vidyanand
Published: February 2005

Aggregation in the cellular slime mold Dictyostelium discoideum is due to chemotaxis. The chemoattractant, cyclic AMP, is synthesised and released periodically by the cells. Externally applied periodic pulses of cyclic AMP can also induce differentiation in this organism. The present work examines the role of periodicity per se in cyclic AMP-mediated stimulation of cell differentiation. For this purpose we use Agip53, a Dictyostelium mutant which does not develop beyond the vegetative state but can be made to aggregate and differentiate by reiterated applications of cyclic AMP. Importantly, Agip53 cells do not make or release any cyclic AMP themselves even in response to an increase in extracellular cyclic AMP. A comparison of the relative efficiencies of periodic and aperiodic stimulation shows that whereas the two patterns of stimulation are equally effective in inducing the formation of EDTA-stable cell contacts, periodic stimuli are significantly superior for inducing terminal differentiation. This suggests that there must be molecular pathways which can only function when stimulation occurs at regular intervals.
Structure and carcinogenicity of Dibenzo(c,g)carbazole derivatives
Authors: Szafarz David , Périn François , Valéro Danièle , Zajdela François
Published: February 2005

The carcinogenicity of several groups of carcinogens is evoked with particular reference to Dibenzo(c,g)carbazole derivatives. The activity of these derivatives is discussed with respect to their species and organ specificity. The enzymatic equipment is decisive as to whether the compounds formed can react with DNA or are simply detoxified and eliminated. All these carcinogens are complete carcinogens, i. e. they have the property of both initiation and promotion.
Protein folding
Authors: Szulmajster Jekisiel
Published: February 2005

The importance of protein folding in the biosynthesis of proteins is reviewed.
Some examples of the functions of metal ions in biology
Authors: Williams R. J. P.
Published: February 2005

The ways in which certain inorganic elements are used in essential roles within biological systems generally are described. The two roles stressed are (1) electrolytic charge movements and (2) electronic charge movement. The value of helical proteins inside and outside membranes as mechanical coupling devices is analysed.
A Unified Matrix Hypothesis of DNA-directed morphogenesis, protodynamism and growth control
Authors: Scherrer Klaus
Published: February 2005

A theoretical concept is proposed, in order to explain some enigmatic aspects of cellular and molecular biology of eukaryotic organisms. Among these are the C-value paradox of DNA redundancy, the correlation of DNA content and cell size, the disruption of genes at DNA level, the “Chromosome field” data of Lima de Faria (Hereditas93:1, 1980), the “quantal mitosis proposition” of Holtzer et al. (Curr. Top. Dev. Biol.7:229, 1972), the inheritance of morphological patterns, the relations of DNA and chromosome organisation to cellular structure and function, the molecular basis of speciation, etc. The basic proposition of the “Unified Matrix Hypothesis” is that the nuclear DNA has a direct morphogenic function, in addition to its coding function in protein synthesis. This additional genetic information is thought to be largely contained in the non-protein coding transcribed DNA, and in the untranscribed part of the genome.
Comparison of liposome entrapment parameters by optical and atomic absorption spectrophotometry
Authors: Yoss Norma L. , Popescu Octavian , Pop Victor I. , Porutiu Dorin , Kummerow Fred A. , Benga Gheorghe
Published: February 2005

Methods for the complete characterization of liposomes prepared by ether-injection are described in detail. The validity of atomic absorption spectrophotometry Ior measuring markers of trapped volume was checked by comparative determinations of markers with established optical spectrophotometrical methods. The favorable results usingl atomic absorption spectrophotometry to quantitate the marker Mn2+ are of particular relevance as manganese ion is also the paramagnetic probe in n.m.r, measurements of water permeability of lipo-somes; our results indicate that in such measurements no other marker need be incorporated.
Biochemical changes induced by Coxsackie B4 virus in short-term culture of mouse pancreatic islets
Authors: Szopa T. M. , Gamble D. R. , Taylor K. W.
Published: February 2005

Isolated mouse pancreatic islets were infected in vitro with two strains of Coxsackie B4 virus – a tissue culture – adapted strain and a mouse pancreas-adapted strain. Within 48 h of infection changes had occurred in the biochemical activities of islets infected with the mouse pancreas-adapted strain of virus. Basal insulin release was increased two-fold in these islets, while glucose-induced insulin secretion remained unchanged. Insulin biosynthesis was greatly reduced at a sti, mulatory concentration of glucose (20 raM), thus leading to a reduced insulin content in these islets. These effects are of importance because they demonstrate that certain strains of Coxsackie B4 virus, like encephalo-myocarditis virus, may selectively alter B-cell function in vitro.
Proteoglycan-type I collagen fibril interactions in bone and non-calcifying connective tissues
Authors: Scott J. E. , Haigh M.
Published: February 2005

The association of proteogtycans with type I collagen fibrils in skin, tendon, cornea and bone has been determined by electron microscopy using an electrondense dye, Cupromeronic blue, in the critical electrolyte concentration mode, backed up by biochemical analysis and digestion by hyaluronidase or keratanase. A major proteoglycan of the soft tissues, containing dermatan sulphat, is shown to be regularly and orthogonally arranged at the surface of the fibrils. Uranyl acetate counterstaining revealed that the main specific binding site is the ‘d’ band, which previous work indicated is very close to the initial site of calcification of type I collagen fibrils. Bone, deminer-alized by a ‘non-aqueous’ technique which preserves the proteoglycan in the tissue does not contain orthogonal arrays; the interfibrillar proteoglycan filaments are oriented parallel to the fibril axis. The main proteoglycan in bone is chondroitin sulphate-rich. It is suggested that dermatan sulphate proteoglycan plays a role in preventing soft connective tissues from calcifying.
Extracellular adhesive molecules in neurite growth
Authors: Rauvala Heikki , Pihlaskari Riitta , Laitinen Jari , Merenmies Jussi
Published: February 2005

This review deals with two topics: (1) the effects of fibronectin and laminin on neurite growth and the molecular mechanisms of these effects, and (2) isolation and properties of the adhesive molecule p30. This novel molecule is an abundant heparin-binding protein in perinatal rat brain, and is suggested to have a role in neuronal growth.
The interaction of prostaglandins with human serum lipoproteins
Authors: Bergelson L. D.
Published: February 2005

Using high density and low density lipoproteins (HDL and LDL) labeled with fluorescent analogues of phosphatidylcholine or sphingomyelin it was found that low amounts (10−12 M) of prostaglandins E1 and F2α induced different structural rearrangements of the lipoprotein surface, whereas prostaglandins E2 and F1α had no effect. The effects of prostaglandin E1 on HDL were largely paralled by those of this prostaglandin on synthetic recombinants prepared from pure apolipoprotein A1, phospholipids and cholesterol and were demonstrated to be caused by prostaglandin-apolipoprotein interaction. The interaction resembled that of a ligand with a specific receptor protein because it was specific, reversible, concentration and temperature dependent and saturable. However the retaining capacity of HDL or LDL for prostaglandin E1 as determined by equilibrium dialysis was very low and a single prostaglandin E1 molecule was able to induce structural changes in large numbers of discrete lipoprotein particles. To explain this remarkable fact a non-equilibrium model of ligand-receptor interaction is proposed. According to that model in open systems characterized by weak ligand-receptor binding, high diffusion rate of the ligand and long relaxation times which exceed the interval between two successive receptor occupations, the ligand-induced changes will accumulate, resulting in transformation of the system into a new state which may be far away from equilibrium. It is emphasized that the low mobility of lipids constituting the environment of the receptor protein plays a critcal role in this type of signal amplification.

It was further demonstrated that the PGE1-induced changes of the lipoprotein surface resulted in an enhancement of LDL-to-HDL transfer of cholesterol esters and phosphatidylcholine especially in the presence of serum lipid transfer proteins. The acceleration of the interlipoprotein transfer caused by prostaglandin E1 in turn increases the rate of cholesterol esterification in serum. It is suggested that in such a way prostaglandin E1 may influence the homeostasis of cholesterol.
Seasonal pattern of glycosylation in frog liver
Authors: Leoni S. , D'Alessandro A. , Conti R. , Marino M. , Spagnuolo S. , Mangiantini M. T.
Published: February 2005

The circannual behaviour of glycosylation and protein synthesis in frog liver slices was studied following the incorporation of3H-galactose and14C-glucosamine into glycolipids and glycoproteins and3H-leucine into proteins. The activity of two enzymes the galactosyl-transferase and the N-acetyl-glucosaminyl-1-P-transferase was determined. The incorporations of both sugars into the soluble fraction and into the lipid extract present a maximum during the spring-summer period. The incorporation into the protein fraction displays a different pattern:14C-Glucosamine and3H-leucine incorporation increases from winter to a maximum in autumn; the incorporation of3H-Galactose has a sharp peak during spring. The pattern of glycosyltransferase activities is similar to the pattern of incorporation of the two saccharides into proteins, indicating these enzymes as important control points for glycosylation in Anurae.
Differences in L-alanine uptake by livers of Wistar and lean Zucker rats
Authors: Ruiz B. , Casado J. , Pastor-Anglada M. , Felipe A.
Published: February 2005

The L-alanine uptake by livers of Wistar and lean Zucker rats has been studied. The hepatic uptake and fractional extraction rates of alanine were estimated in 50–55 day old rats. No significant differences in amino acid concentrations and blood flows in afferent and efferent liver vessels were seen in lean Zucker rats when compared with Wistar rats. However, the hepatic uptake (1.6±0.1 and 0.7±0.1 μmol/min/100 g bw, p<0.01) and the fractional extraction (26.8±2.1 and 15.2±3.1%, p<0.05) were much lower in Zucker than in Wistar rats. The hepatic active transport of L-alanine was determined in vitro using isolated plasma membrane vesicles. Vesicles isolated from livers of lean Zucker rats showed similar values of Km (2.5±0.7 vs 2.0±0.5 mM for Wistar and Zucker respectively, N.S.), but lower values of Vmax when compared with Wistar rats (1.1±0.1 vs 0.6±0.005 nmol/mg prot 5 s, p<0.01, for Wistar and lean Zucker rats respectively). These results indicate that, the liver of lean Zucker rats concentrates alanine less efficiently than the liver of Wistar rats. This fact correlates well with a lower capacity of the Na+-dependent L-alanine trasport in liver plasma membrane vesicles from lean Zucker rats.
Nyctohemeral rhythmicity of type II thyroxine 5′-deiodinase activity in the pineal gland but not in the Harderian gland of the Swiss mouse
Authors: Rubio A. , Osuna C. , Lopez-Gonzalez M. A. , Reiter R. J. , Guerrero J. M.
Published: February 2005

Type II thyroxine 5′-deiodinase (5′-D) activity in both pineal and Harderian glands of the Swiss mouse was studied. Pineal 5′-D activity exhibited a nyctohemeral profile with a maximal peak value at 05.00 h, which coincides with that for pineal melatonin production. However, no rhythm of 5′-D activity in the Harderian gland could be found. In pineal gland, light at night inhibited the nocturnal increase in 5′-D activity, while isoproterenol, a β-adrenergic agonist, could not stimulate the enzyme. In the Harderian gland, neither darkness, nor light at night, or isoproterenol were capable of modifying basal values of 5′-D activity.
Protein synthesis is activated in primed neutrophils: a possible role in inflammation
Authors: Hughes Valerie , Humphreys John M. , Edwards Steven W.
Published: February 2005

Circulating human neutrophils exhibited low rates of protein biosynthesis, as determined by their ability to incorporate [35S]methionine into TCA-precipitable material. Exposure of cells to the chemotactic peptide (N-formyl-L-methionyl-L-leucyl-L-phenylalanine) increased their rate of protein synthesis, and the maximal stimulation of biosynthesis by this inflammatory agent was observed at 0.1 μM: this concentration of chemotactic peptide “primed” neutrophil activity and only activated the oxidase of these cells by 8% of maximum. The newly-synthesized proteins were analyzed by two-dimensional polyacrylamide gel electrophoresis and compared with those synthesized in control cells. Two classes of proteins were observed in “primed” cells. The first of these comprised proteins whose rate of biosynthesis changed very little upon “priming” whereas the second class comprised proteins whose rate of synthesis increased greatly after exposure to chemotactic peptide. The fMet-Leu-Phe stimulated protein synthesis was inhibited by actinomycin D and cycloheximide showing that this phenomenon required both transcription and translation. We propose that these fMet-Leu-Phe regulated proteins play an important role in the function of neutrophils during an inflammatory response.
Hormone-sensitive lipase in brown adipose tissue: Identification and effect of cold exposure
Authors: Holm Cecilia , Fredrikson Gudrun , Cannon Barbara , Belfrage Per
Published: February 2005

Hormone-sensitive lipase (HSL) in brown adipose tissue from mice was identified through immunoprecipitation with a polyclonal antibody (anti-HSL) towards rat white fat HSL and Western blotting. An 82 kDa polypeptide, slightly smaller than the rat white fat HSL 84 kDa subunit, was detected and its identity as HSL verified by inhibition properties. The HSL concentration per g tissue was several-fold higher in the mouse brown adipose tissue than in the rat white adipose tissue, but the specific activities per mg protein were similar. Cold-exposure (4°C of the mice for 24 h approximately doubled the HSL concentration but this increase parallelled the overall protein increase and did not reflect a specific effect on the HSL.
The effect of tunicamycin on transferrin synthesis and secretion in hormonally stimulated explants of rabbit mammary gland
Authors: Bradshaw Judith P. , White David A.
Published: February 2005

Pregnant rabbit mammary gland explants cultured with insulin, prolactin and cortisol synthesize and secrete transferrin radiolabelled with [3H]leucine or [ 3H]mannose. Addition of tunicamycin inhibits the glycosylation but not synthesis of the [3H]leucine-labelled poly-peptide. Secretion of the non-glycosylated protein is only marginally inhibited.
Further studies on the presumed adenine nucleotide storage compound of rat heart
Authors: Fitt Peter S. , Korecky Borivoj , Sharma Nishi
Published: February 2005

Further studies on the acid-precipitable radioactive substance formed during perfusion of Langendorff rat hearts with [14C]adenosine have shown that very brief (30 s) ischaemia causes a sudden rise (20–35%) in its level in the tissue which is followed by the steady fall we have previously described. Analysis of the products of alkaline hydrolysis of this compound shows that at least 96% of the radioactivity appears in the form of a mixture of 2′- and 3′-AMP as would be expected for RNA while its relatively high resistance to dilute alkali suggests that it is poly A. Subcellular localization studies indicate that radioactivity enters all compartments of the cell, with maximum label in the nucleus. However, a significant proportion is present in the mitochondria and may be poly A acting as the mitochondrial storage form of adenine nucleotides whose existence we have proposed.
Activation of transglutaminase at calcium levels consistent with a role for this enzyme as a calcium receptor protein
Authors: Hand D. , Bungay P. J. , Elliott B. M. , Griffin M.
Published: February 2005

The sensitivity of tissue transglutaminase to activation by Ca2+ and other cellular factors was investigated using the enzyme purified from rat liver. The inclusion of Mg2+ in the assay system appeared to reduce the Ca2+-requirement of the enzyme when native N,N′-dimethylcasein was used as the protein acceptor substrate. However, when this protein was dephosphorylated, the Ca2+-requirement was unaffected by Mg2+. In addition, using this modified assay, a Km for Ca2+ was calculated to be in the range of 3–4 μM, at least an order of magnitude lower than that obtained with native acceptor substrate. Membrane phospholipids, 1,2-diolein and calmodufin were found not to affect the activation oftransglutaminase by Ca2+. The sensitivity of transglutaminase to Ca2+ which we have now demonstrated suggests that this enzyme may directly act as a receptor protein for Ca2+ during stimulusresponse coupling mediated by this cation.
Immobilization of enzymes masks their active site
Authors: Kumakura Minoru , Kaetsu Isao
Published: February 2005

We studied the effect of immobilizing cellulase to carboxycellulose sodium by radiation polymerization on the masking of the active site of the enzyme. Masking of the enzyme during the preparation of immobilized enzyme was assayed at tow temperature. The activity of immobilized enzyme was retained during repeated batch reactions, indicating that the enzyme was firmly trapped in the polymer matrix. Various compounds (designated monomers) were used to dissolve the carboxymethylcellulose; enzyme activity was affected by the nature of the monomer, by the monomer concentration, and by the solubility of the substrate in monomer.
Measurement of anti-glomerular-basement-membrane antibodies by micro ELISA using insoluble antigens
Authors: Barnard Kay , James M. P.
Published: February 2005

An enzyme immunoassay, using finely ground rabbit glomerular basement membrane (GBM) as an antigen, was able to detect sheep anti-rabbit GBM antibodies at serum dilutions of 1:32 000. The particulate GBM bound firmly to plastic micro ELISA plates without the aid of a linking agent, and the antigen-coated plates remained stable for several months when stored at −70°C. There were no appreciable differences between amino acid compositions of ground and whole GBM, and no detectable loss of antigens occurred during the grinding procedure. Competitive inhibition assays, with collagenase and pepsin digests of rabbit GBM demonstrated preservation of collagenous and non-collagenous antigens in the ground GBM. The assay should prove to be a relatively simple and highly sensitive technique for detecting antibodies to a wide spectrum of GBM antigens.
An inhibitor of mannosylation of retinyl-phosphate
Authors: Datema Roelf , Schwarz Ralph T.
Published: February 2005

The guanosine disphospate and uridine diphosphate esters of the antiviral sugar analog 2-deoxy-2-fluoro-D-glucose (GDP-FGlc and UDP-FGlc) were synthesized and tested as inhibitors of formation of lipid-linked sugars in cell-free extracts. Formation of dolichol-phosphate mannose and of dolichol-diphosphate di-N-acetylchitobiose were not inhibited by either sugar nucleotide. Formation of dolichol-phosphate glucose was inhibited by UDP-FGlc, not by GDP-FGlc. Although GDP-FGlc did not inhibit formation of dollchol-phosphate mannose, it did inhibit formation of retinol-phosphate mannose from retinol-phosphate and GDP-Man. This inhibition was not reversed by exogenous retinol-phosphate, nor was FGIc from GDP-FGlc incorporated into retinolphosphate-linked derivatives. As FGLc inhibits formation of dolichol-phosphate mannose in intact cells, but does not decrease pool sizes of GDP-Man and dolichol-phosphate (Datema et al., 1980, Eur. J. Biochem. 109, 331–341), we discuss that inhibition of formation of retinol-phosphate mannose by one of the metabolites of FGlc, namely GDP-FGlc, may lead to decreased synthesis of dolichol-phosphate mannose in FGlc-treated cells. This implies a role for vitamin A in the dolichol cycle of protein glycosylation.
Divergent neuropeptide evolutionary drifts between American and Australian marsupials
Authors: Chauvet J. , Hurpet D. , Chauvet M. T. , Acher R.
Published: February 2005

Present-day marsupials, which are supposed to have arisen from a single stem diverging from the placental stem some 130 million years ago, exist only in the American and Australian continents. Comparison of the homologous genes and their protein products, which evolved under different environmental conditions, may provide arguments for either selective or neutral evolution. In contrast to Australian Macropodidae, which have peculiar neurohypophysial peptides, namely mesotocin and two pressor peptides, lysine vasopressin and phenypressin, the South American oppossum, Didelpbis marsupialis, has oxytocin, lysine vasopressin, and arginine vasopressin. Because placental mammals have oxytocin and usually arginine vasopressin, and nonmammalian tetrapods have mesotocin and arginine vasotocin, it is assumed that (1) selective change of arginine vasotocin into arginine vasopressin occurred in mammalian ancestors and a subsequent gene duplication in the marsupial line gave rise to two pressor peptides with divergent neutral drifts in American and Australian groups, and (2) mesotocin of nonmammalian tetrapods has been preserved in Australian marsupials and reclaimed for milk-ejecting function whereas it has been converted into oxytocin in South American oppossums. The change of mesotocin into oxytocin seems neutral rather than selective.
Effect of calcium and F-actin on the rate of SH2 modification in skeletal myosin
Authors: Kameyama Tsuneo , Ishii Mitsuko K. , Yoshiike Takashi
Published: February 2005

The rate constant of modification of a specific thiol group, SH2, with N-ethylmaleimide (NEM) has been used to estimate the conformational change in the local area containing SH2 (SH2 region) of skeletal myosin as a structural probe. The rate of Mg2+-ATP-induced SH2 modification of subfragment-1 (S-l) isozymes was regulated by Ca2+ in the pCa range below 6.4 and was not regulated in the pCa range above 6.4. No substantial difference between S-1 containing alkali light chain, A1, (S-1(A1)) and S-1 containing alkali light chain, A2, (S-1(A2)) was observed in the Ca2+-dependent rate of SH2 modification. Due to the presence of this Ca2+ regulation in myosin (absence in S-1 isozymes) in the pCa range above 6.4, absence of 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) light chain in S-1 isozymes, and high affinity of Ca2+ for DTNB light chain, this Ca2+ regulation in the pCa range above 6.4 is possibly related to the Ca2+ binding to DTNB light chain. F-Actin, which is entirely free from tropomyosin and troponin, enhanced the rate of Mg2+-ATP-induced SH2 modification of S-1 isozymes equally and of myosin, and reduced the Ca2+ sensitivity with an increase in F-actin concentration.
Mitochondrial pyruvate carrier—A possible link between gluconeogenesis and ketogenesis in the liver
Authors: Kümmel Ladislav
Published: February 2005

Effects of various ketogenic substrates on gluconeogenesis from lactate or alanine were compared. The results suggest that, in intact liver cells, cytoplasmic pyruvate is transported into mitochondria in exchange for intramitochondrially generated acetoacetate. An interrelationship between gluconeogenesis and ketogenesis may thus exist in the liver at the level of mitochondrial pyruvate carrier.