bluuebunny/crossref_metadata_2025 huggingface dataset:
Sequences isolated from aDrosophilaearly-ecdysone puff are expressed in rat liver
Authors: Arribas Carmen , Izquierdo Marta
Published: February 2005

We have studied the presence of a cloned fragment of DNA from Drosophila melanogaster in other organisms by means of nucleic acid hybridization analysis. The isolated region is localized in polytene chromosomes at the 63F subdivision. This region includes a puff that responds within minutes to ecdysone stimulation. We have found that 63F DNA from D. melanogaster hybridizes ‘in situ’ to both DNA and RNA from D. simulans, D. teissieri, and D. hydei. In all these species the isolated DNA remains associated with one early-ecdysone stimulated puff. The isolated Drosophila recombinant DNA is also complementary to polyadenylated RNA from foetal and adult rat liver but fails to hybridize to the nonpolyadenylated RNA classes from both sources and to polyadenylated RNA from rat mammary glands.
Preparation of hormone-sensitive adipocytes from foetal rat brown adipose tissue
Authors: Elliott Keith R. F. , Roncero Cesar , Fernandez Margarita , Benito Manuel
Published: February 2005

A collagenase-dispersion method involving Percoll gradient separation is described for the production of adipocytes from foetal rat brown adipose tissue. The cells produced are hormone-sensitive as judged by cAMP accumulation and lipolysis.
Uncoupling and isotope effects in γ-butyrobetaine hydroxylation
Authors: Holme Elisabeth , Lindstedt Sven , Nordin Ingalill
Published: February 2005

Replacement of unlabeled γ-butyrobetaine with γ-[2,3,4-2H6]butyrobetaine has a profound effect on the stoichiometry between decarboxylation of 2-oxoglutarate and hydroxylation in the reaction catalyzed by human γ -butyrobetaine hydroxylase. The ratios between decarboxylation and hydroxylation are 1.16 with Unlabeled and 7.48 with deuterated γ-butyrobetaine as substrate. From these ratios an internal isotope effect of 41 has been calculated. DV in the overall reaction measured as 2- oxoglutarate decarboxylation is 2.5 and DV/K is 1.0. For γ-butyrobetaine hydroxylase from Pseudomonas sp. AK 1, 2-oxoglutarate decarboxylation exceeds hydroxylation with 10% when deuterated γ-butyrobetaine is used. No excess was found with unlabeled substrate and no internal isotope effect could be calculated. DV for the bacterial enzyme is 6.
The matrix water space of mitochondriain situin isolated hepatocytes
Authors: Lund Patricia , Wiggins David
Published: February 2005

The intracellular and mitochondrial matrix water-permeable spaces were measured in intact, respiring hepatocytes from normal fed, 48 h starved, and protein fed rats. The mitochondrial matrix constitutes 15–20 % of the total intracellular water, a proportion greater than has generally been assumed.
Elementary steps in synaptic transmission revealed by currents through single ion channels
Authors: Sakmann Bert
Published: February 2005

An account is presented of how the molecular basis of synaptic transmission at peripheral and central synapses is elucidated by combining patch clamp and recombinant DNA techniques.
On the signal transducing mechanisms involved in the synergistic interaction between interleukin-1 and bradykinin on prostaglandin biosynthesis in human gingival fibroblasts
Authors: Lerner Ulf H. , Brunius Gustaf , Modeer Thomas
Published: February 2005

Recombinant human interleukin-1β (IL-1β) and bradykinin (BK) synergistically stimulate prostaglandin E2 (PGE2) formation in human gingival fibroblasts cultured for 24 h. Neither BK or IL-1β per se, nor their combinations, caused any acute stimulation of cellular cyclic AMP accumulation. BK, but not IL-1β, caused a rapid, transient rise of intracellular Ca2+ concentration ([Ca2+]i), as assessed by recordings of fura-2 fluorescence in monolayers of prelabelled gingival fibroblasts. IL-1β did not change the effect of BK on [Ca2+]i. Ionomycin and A 23187, two calcium ionophores, synergistically potentiated the stimulatory effect of IL-1β on PGE2 formation. Three different phorbol esters known to activate protein kinase C also synergistically potentiated the action of IL-1β on PGE2 formation. Exogenously added arachidonic acid significantly enhanced the basal formation of PGE2. In IL-1β treated cells, the enhancement of PGE2 formation seen after addition of arachidonic acid, was synergistically upregulated by IL-1β. These data show that i) the synergistic interaction between IL-1β and BK on PGE2 formation is not due to an effect linked to an upregulation of cyclic AMP or [Ca2+]i; ii) the signal transducing mechanism by which BK interacts with IL-1β, however, may be linked to a BK induced stimulation of [Ca2+]i and/or protein kinase C; iii) the mechanism involved in the action of IL-1β may, at least partly, be due to enhancement of the biosynthesis of prostanoids mediated by an upregulation of cyclooxygenase activity.
In vitro secretion of transforming growth factor alpha (TGF-α): A comparison of the A431 cell line with three human oesophageal squamous cell carcinoma lines
Authors: Thornley A. L. , Jones G. J.
Published: February 2005

Transforming growth factor alpha (TGF-α) is a single chain polypeptide which exists in a variety of forms differing in molecular weight. These forms are variously present in normal and neoplastic cells. Of particular interest are TGF-α's well-known mitogenic properties. The transition from a normal to a neoplastic cellular state results from signalling defects that may depend upon, iter alia, abonormal levels of expression and secretion of TGF-α. It is known that the secretion of TGF-α may be enhanced appreciably by agents such as phorbol 12-myristate 13-acetate (PMA), serum factors and epidermal growth factor (EGF). Here, we compare the efficacy of these three agents in the elevation of TGF-α secretion in the well studied A431 cell line with their previously undocumented efficacy in certain interesting, but little known, human oesophageal squamous cell carcinoma (SCC) lines.
Lindane-induced modifications to membrane lipid structure: Effect on membrane fluidity after subchronic treament
Authors: Gutierrez-Ocaña M. T. , Senar S. , Perez-Albarsanz M. A. , Recio M. N.
Published: February 2005

Chronic lindane intoxication by injecting subcutaneously the toxicant, resulted in an altered lipid pattern in rat ventral prostate membranes. An increase of membrane fluidity was also observed using a fluorescence polarization technique. When in vitro experiments were carried out with both treated and untreated rats, an interesting lack of parallelism was found, which could indicate the development of a resistance to membrane disordering by lindane. The observed changes in cholesterol and phospholipid composition are also consistent with the hypothesis that lindane perturbs the lipid matrix of membranes, possibly inducing complex compensatory changes in the membrane lipid composition.
Co-operative nature of N-glycosylation of proteins at multiple sites: evidence from studies with tunicamycin
Authors: Cushley W. , Singer H. H. , Williamson A. R.
Published: February 2005

The pattern of glycosylation of newly synthesized human and routine immunoglobulin μ-chains varies according to the degree of inhibition of N-glycosylation effected by using the antibiotic tunicamycin. Tunicamycin, at high concentrations, can apparently block N-glycosylation of μ-chains completely as judged by SDS-PAGE analysis of the biosynthetically labelled products. At lower concentrations of tunicamycin, fully giycosy]ated and totally non-glycosylated chains were the predominant molecular species. The paucity of the predicted partially glycosylated μ-chains leads us to suggest that the addition of oligosaccharides to the appropriate acceptor sites is a cooperative process.

Long exposure of fluorographs reveals each of the predicted intermediately glycosylated forms of the μ-chain, and counting the number of bands on such fluorographs may prove useful in the preliminary determination of the number of N-linked oligosaccharides in a given glycoprotein.
A staining method for the detection of impaired photosynthetic electron transport in green algae strains
Authors: Dauta Jacques
Published: February 2005

The utilization of strain CW 15 of Chlamydomonas reinhardtii enables the biochemical staining test for characterizing ad efective photosynthetic electron-transport chain on colonies grown on agar medium. It may be used in order to screen for mutants specifically blocked in different parts of the transport chain.
Solid-phase enzyme immunoassay of urokinase using monoclonal antibodies
Authors: Hérion P. , Portetelle D. , Franssen J. -D. , Urbain J. , Bollen A.
Published: February 2005

Using two monoclonal antibodies directed against urokinase, we have developed a micro enzyme-linked immunosorbant assay (ELISA) to detect and measure urokinase in biological fluids. The system presents the following characteristics: simple and rapid procedure, reproducibility, sensitivity (urokinase levels down to 1 ng/ml) and evaluation of the enzyme in biological fluids such as urine, pleural elfusions, and ascitic fluids without preliminary purification.
Characterization of specific calcitonin gene related peptide receptors present in hamster pancreatic β cells
Authors: Barakat A. , Rosselin G. , Marie J.-C.
Published: February 2005

Calcitonin gene-related peptide (CGRP) shares about 46% and 20% amino acid sequence homology with islet amyloid polypeptide (IAPP) and salmon calcitonin (sCT). We investigated whether these related peptides could cross-react with the specific binding of 125I-[His]hCGRP I to the CGRP receptor in hamster insulinoma cell membranes. A rapid dissociation of membrane bound 125I-[His]hCGRP I could be induced in the presence of 1 μM chicken CGRP (cCGRP). The specific 125I-[His]hCGRP I binding was inhibited by the related peptides and their half-maximal inhibitory concentrations (IC50) were: cCGRP (0.1 nM), rat CGRP I and human CGRP I and II (1.0–2.0 nM), fragment of hCGRP I (8–37) (150 nM), human IAPP (440 nM). The non-amidated form of hIAPP; human diabetes-associated peptide (hDAP) did not inhibit the binding of 125I-[His]hCGRP I and sCT was only effective at a high concentration (1 μM). Binding of 125I-[His]hCGRP I was dose dependently inhibited by guanosine-5′-O-(3-thiotriphosphate) or (GTPγS) and a 70% reduction of binding was obtained with 0.1 mM GTPγS. The IC50 value of cCGRP (0.1 nM) was increased 100-fold in the presence of 0.1 mM GTPγS. Human CGRP I and cCGRP at 2.5 μM did not stimulate the activity of hamster insulinoma cell membranes adenylate cyclase, while glucagon (1 μM) induced a 2-fold increase. Thus, specific CGRP receptors present in hamster β cells are associated with G protein (s) and IAPP can interact with these receptors. These results and the observation that cCGRP and hCGRP I did not influence adenylate cyclase activity provide further evidence for CGRP receptor subtypes.
CO-reacting haemoproteins of neutrophils: Evidence for cytochrome b-245 and myeloperoxidase as potential oxidases during the respiratory burst
Authors: Edwards Steven W. , Lloyd David
Published: February 2005

Room temperature, CO-difference spectra of intact rat polymorphonuclear leucocytes (neutrophils) revealed the presence of a number of CO-binding haemoproteins. Absorption maxima at 413, 540 and 570 nm were attributed to the CO-complex of cytochrome b-245 whereas an absorption maximum at 595 nm was assigned to the contribution from a myeloperoxidase complex, since an identical absorption maximum was observed in CO-difference spectra of purified myeloperoxidase in the presence of H2O2. Photochemical action spectra for the relief of CO-inhibited O2 uptake revealed contributions from both cytochrome b-245 and myeloperoxidase. The potential of these two O2- and CO-binding haemoproteins to function as oxidases during the respiratory burst is discussed.
Degradation artefacts during sample preparation for sodium dodecyl sulphate polyacrylamide gel electrophoresis
Authors: Cookson E. Jane , Beynon Robert J.
Published: February 2005

Preparation of samples for sodium dodecyl sulphate polyacrylamide gel electrophoresis routinely involves heating the protein in solution containing detergent and reducing agent for at least two minutes. Here we show that this treatment causes fragmentation of the protein glycogen phosphorylase, whether purified or as a component of a skeletal muscle preparation. The fragments are detected as minor bands on western blots and represent the products of discrete breakage point in the peptide sequence. Protease inhibitors cannot suppress the fragmentation.

Such small amounts of immunoreactive fragments may be incorrectly identified on western blots as contaminants that were originally present in the antigen preparation. They may also be a source of ambiguity in studies that search for degradation intermediates during proteolysis.
Acetoacetyl-CoA ligase activity in the isolated rat hepatocyte: Effects of 25-hydroxycholesterol and high density lipoprotein
Authors: Wong Gail A. , Bergstrom James D. , Edmond John
Published: February 2005

The activity of acetoacetyl-CoA (AcAc-CoA) ligase (E.C.6.2.1.16) in hepatocytes from rats was shown to be the same as the activity in homogenates of their livers. In hepatocytes treated with 25-hydroxycholesterol, AcAc-CoA ligase, 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) reductase and rates of sterol synthesis were substantially decreased. Hepatocytes treated with high density lipoprotein (HDL) exhibited a 2 to 4 fold induction of HMG-CoA reductase activity; however an accompanying increase in AcAc-CoA ligase activity and the rate of cholesterol synthesis was not observed. We conclude (a) that increases in the activity of HMG-CoA reductase when mediated by HDL in hepatocytes do not result in a corresponding change in the capacity for sterol synthesis and (b) that changes in the activity state of HMG-CoA reductase can be dissociated from that of AcAc-CoA ligase.
Purification of four gelatin-binding proteins from bovine seminal plasma by affinity chromatography
Authors: Manjunath P. , Sairam M. R. , Uma J.
Published: February 2005

Bovine seminal plasma contains three similar acidic proteins, which we have previously designated as BSP-A1, BSP-A2, and BSP-A3. These proteins contain two homologous domains that are similar to type II structures present in the gelatin-binding domain of fibronectin. The present data have revealed that these proteins, like fibronectin, also form complexes with gelatin, a denatured collagen. Based on this property, a single step affinity purification method has been developed. In addition to these three proteins BSP-A1, −A2 and −A3, another protein with an apparent molecular weight of 30,000 dalton (named BSP-30-kDa) also bound to the gelatin-agarose column. Elution of these proteins from affinity columns using a linear gradient of either urea or arginine gave essentially the same pattern with a high yield of 90–95%. The purified proteins were homogeneous by SDS-polyacrylamide gel electrophoresis, amino acid composition and HPLC. Chromatography of bull seminal vesicular fluid also exhibited an elution pattern similar to that obtained for bull seminal plasma. The availability of these purified proteins should aid in understanding the physiology of these gelatin-binding proteins.
On the specificity of antisense RNA to arrest in vitro translation of mRNA coding for Drosophila hsp 23
Authors: Nicole Louis M. , Tanguay Robert M.
Published: February 2005

The specificity of action of antisense RNA for one of Drosophila low molecular weight heat shock proteins (hsp 23) was tested at the translational level using the rabbit reticulocyte lysate cell-free system. T7 polymerase-driven transcripts of hsp 23 in the antisense orientation were mixed with mRNA from heat-shocked cells under various stringency conditions prior to translation in vitro. Although the four small hsps show considerable sequence homology in their coding sequences, antisense hsp 23 RNA was shown to specifically inhibit hsp 23 mRNA translation under both high (formamide, 45°C and low stringency (37°C conditions. This suggests that the 5′ leader and the ribosome binding region of mRNA are of prime importance in the specificity of action of antisense RNA at the translational level.
Regulation of liver base-exchange activity by acidic phospholipids
Authors: Zborowski Józef , Corazzi Lanfranco , Arienti Giuseppe
Published: February 2005

Liver microsomes were enriched in liposomal acidic lipids by Ca2+-dependent fusion of liposomes at pH 7.0. The extent of fusion was monitored by the transfer of radioactive cholesteryl oleate. The enrichment of membranes in phosphatidylserine inhibited ethanolamine base-exchange, whereas the fusion with phosphatidylinositol inhibited both ethanolamine and serine base-exchange reactions. In contrast, these two phospholipids had scarce effects on choline base-exchange. Phosphatidic acid did not suppress any of the three base-exchange activities. Possible functional implications are discussed.
Chick hepatic lectins: An electron microscopic study on isolated hepatocytes during development
Authors: Falasca L. , Mattioli P. , Dini L.
Published: February 2005

We studied the carbohydrate recognition systems of hepatocytes isolated from 16-day-old embryos, 19-day-old embryos and chicks within 24 h of hatching. We localized and quantified at the ultrastructural level the binding sites for glycoproteins exposing terminal N-acetylglucosamine (GlcNAc), mannose and N-acetylgalactosamine (GalNAc) residues by means of protein-gold complexes. Binding sites specific for GlcNAc and mannose residues are present on hepatocytes from embryos and chicks. On the contrary GalNAc specific binding sites are exclusively observed on cells from 16-day-old embryos. The number and distribution of gold particles on hepatocyte cell surfaces depend on the binding sites and the age considered. We describe a modulation in the number of GlcNAc, and mannose specific receptors present on the cell surface between the embryonal stage and neonatal life.
Binding of doxorubicin to cardiolipin as compared to other anionic phospholipids—An evaluation of electrostatic effects
Authors: de Wolf Frits A.
Published: February 2005

The binding of doxorubicin to large unilamellar vesicles consisting of cardiolipin or other anionic phospholipids was analyzed in terms of the local drug concentration at the membrane surface, according to the Gouy-Chapman theory. The analysis suggests strong positive binding cooperativity. Part of the drug binds in the uncharged form. The affinity for cardiolipin and other anionic phospholipids is comparable. A binding level of 0.5 doxorubicin per lipid-phosphorus is reached when the local concentration of free doxorubicin monomer-equivalents at the membrane surface is about 0.2–0.7 mM. This contrasts with earlier findings indicating a 300–1000 fold higher affinity for cardiolipin. The present analysis provides an explanation for this apparent discrepancy.
Glucose handling by hepatocytes from obese Zucker rats
Authors: Carbó Neus , López-Soriano Francisco J. , Argilés Josep M.
Published: February 2005

Hepatocytes isolated from obese Zucker rats showed a significantly higher rate of both [U-14C]glucose and [U-14C]lactate incorporation into [14C]lipid than those from their lean counterparts. This was associated with a marked increase in the lipogenic rate measured by the incorporation of3H2O into the cell esterified fatty acids. Although there were no changes in the incorporation of the tracer into either [14C]glycogen or14CO2, the [14C] total uptake was significantly higher in the obese animals. The high rate of [14C]lipid synthesis from glucose was observed both at 15 and 30 mM substrate concentrations and was linked to an enhanced uptake of the tracer into the cell as measured using the decarboxilation of [1-14C]glucose in the presence of phenazine methosulphate. The presence of insulin in the incubation medium had no effect on the uptake of glucose by the liver cells. However, the large uptake of glucose by the hepatocytes from the obese animals was not related to an enhanced rate of transport as measured using 3-O-methyl[U-14C]glucose. The activity of glucose-6-phosphate dehydrogenase together with a higher [1-14C]glucose/[U-14C]glucose descarboxylation ratio indicate a predominant very active pentose phosphate pathway which may be responsible for the enhanced glucose uptake observed in the hepatocytes from the obese animals.
Effect of alkylation with N-methyl-N-nitrosourea and N-ethyl-N-nitrosourea on the secondary structure of DNA
Authors: Rizvi R. Y. , Hadi S. M.
Published: February 2005

We have earlier reported that alkylation of DNA by the chemical carcinogen dimethyl sulphate, which mainly alkylates N-7 of guanine and N-3 of adenine, causes the formation of partially denatured regions in double-stranded DNA (Rizvi RY, Alvi NK & Hadi SM, Biosci. Rep.2, 315–322, 1982). It is known that the major site of alkylation in DNA by N-ethyl-N-nitrosourea (EtNu) are the phosphate groups. N-methyl-N-nitrosourea (MeNu), on the other hand, causes the alkylation of mainly guanine residues. We have therefore studied the effect of these two alkylating carcinogens on the secondary structure of DNA. DNA aikylated with increasing concentrations of EtNu and MeNu was subjected to alkaline and S1 nuclease hydrolysis. Thermal melting profiles of alkylated DNA were also determined using S1 nuclease. The results indicated that alkylation by the two alkylating agents had a differential effect on the secondary structure of DNA. EtNu-alkylated DNA was found to be more thermostable than native DNA at neutral pH. It was however more alkali-labile than MeNu-alkylated DNA. The greater stability of EtNu-alkylated DNA was considered to be due to abolition of negative charges on phosphate alkylation.
Effects of monensin on biosynthesis and intracellular processing of proinsulin
Authors: Smith J. E. , Tyhurst M. , Howell S. L.
Published: February 2005

Monensin, a specific sodium ionophore, has been shown to reduce glucose-induced proinsulin biosynthesis by 30% and to completely inhibit the intracellular conversion of proinsulin to insulin. Autoradiography of monensin-treated cells demonstrated the presence of large quantities of newly synthesized proteins in amorphous vesicles close to the Golgi complex of B ceils. The results suggest profound effects of monensin on biosynthesis and intracellular processing of proinsulin.
Methylglyoxal inhibits the translation of natural and chemically decapped mRNAs
Authors: Lozano Andres M. , Mezl Vasek A.
Published: February 2005

Methylglyoxal was a weak inhibitor of translation in the reticulocyte-lysate cell-free system and it did not display cap-dependent inhibition. A similar inhibition was obtained in a wheat-germ cell-free system that displayed extensive cap-dependent inhibition with the cap analogue 7-methylguanosine phosphate. These results show that the chemical reaction of methyl-glyoxat with 7-methylguanosine is not the mechanism for the inhibition of protein synthesis by methylglyoxal and that methylglyoxat is a weak general inhibitor of translation.
A monoclonal antibody (HNo-G7) with specificity for a human nucleolar protein
Authors: Shallal Asaad , Campbell Ailsa M.
Published: February 2005

A murine monoclonal antibody which reacts strongly with the nucieoli of human epithelial cells has been isolated. The antibody is of the IgM class and the antigen has a molecular weight of 45 000. The antibody appears to react with interphase chromatin only and to have specificity for epithelial cells.
Defective regulation of apical membrane chloride transport and exocytosis in cystic fibrosis
Authors: McPherson M. A. , Shori D. K. , Dormer R. L.
Published: February 2005

A biochemical link is proposed between recent observations on defective regulation of Cl− transport in CF respiratory epithelial cells and studies showing altered biological activity of calmodulin in exocrine glands from CF patients. A consensus is emerging that defective β-adrenergic secretory responsiveness in CF cells is caused by a defect in a regulator protein at a site distal to cyclic AMP formation. Our results indicate that this protein might be a specific calmodulin acceptor protein which modifies the activity of calmodulin in epithelial cells. Alteration in Ca2+/calmodulin dependent regulation of Cl− transport and protein secretion could explain (i) alterations in Ca2+ homeostasis seen in CF, (ii) defective β-adrenergic responses of CF cells, and (iii) the observed inability of cyclic AMP (acting via its specific protein kinase, A-kinase) to open apical membrane Cl− channels in CF epithelial cells. Most of the physiological abnormalities of CF including elevated sweat electrolytes and hyperviscous mucus can be explained on this basis.
The relationship between the bilayer to hexagonal phase transition temperature in membranes and protein kinase C activity
Authors: Epand Richard M. , Stafford Alan R. , Cheetham James J. , Bottega Remo , Ball Eric H.
Published: February 2005

A number of substances affect the activity of protein kinase C. Among uncharged and zwitterionic compounds, those which activate protein kinase C also lower the bilayer to hexagonal phase transition temperature of dielaidoylphosphatidylethanolamine while substances which inhibit protein kinase C raise this transition temperature. Using this criteria, we have identified 3β-chloro-5-cholestene, 5β-cholan-24-ol and eicosane as new protein kinase C activators and have shown that Z-Ser-Leu-NH2, Z-Gly-Leu-NH2, Z-Tyr-Leu-NH2, cyclosporin A and cholestan-3β, 5α, 6β-triol are protein kinase C inhibitors.
Cytosolic calcium, oxygen consumption and the intracellular pH of stimulated neutrophils
Authors: Nasmith Patricia E. , Grinstein Sergio
Published: February 2005

The cytoplasmic pH undergoes a biphasic change when neutrophils are activated. The role of Ca2+ in initiating these changes was investigated. No correlation was found between the increased cytosolic [Ca2+] and the stimulation of the Na+/H+ antiport. Similarly, the cytoplasmic acidification elicited by activation in Na+-free media was found to be unrelated to [Ca2+]. Reversal of Na+/H+ exchange was also ruled out as the source of the acidification. Data using a variety of soluble activators indicate that metabolic acid generation is largely responsible for the observed drop in cytoplasmic pH.
Bovine corneal stroma contains a structural glycoprotein located in the gap region of the collagen fibrils
Authors: Wall Rita S. , Elliott Gerald F. , Gyi Tracy J. , Meek Keith M. , Branford-White Christopher J.
Published: February 2005

Treatment of bovine corneal stroma using SDS-containing extracting solutions removes a 135,000 MW glycoprotein from the main collagen framework of the tissue. Low-angle synchrotron X-ray diffraction patterns obtained from corneas extracted in this way indicate that the glycoprotein has been removed from the gap regions of the collagen fibrils and is thus an important structural component of the corneal stroma. The glycoprotein (GP 135) shares a number of properties with one of the subunits of type VI collagen, but tests have so far failed to establish their identity.
Methyl gallate, methyl-3,4,5-trihydroxy-benzoate, is a potent and highly specific inhibitor of herpes simplex virus in vitro. I. Purification and characterization of methyl gallate from Sapium sebiferum
Authors: Kane Cynthia J. M. , Menna Jay H. , Yeh Yun-Chi
Published: February 2005

A potent anti-herpetic compound was identified and purified to homogeneity from the leaves of Sapium sebiferum by plaque reduction assay using herpes simplex virus type 2. The chemical structure of the purified compound was determined by mass spectroscopy and proton and carbon-13 nuclear magnetic resonance as methyl gallate, methyl-3,4,5-trihydroxybenzoate. This is the first demonstration that methyl gallate is a potent anti-herpetic compound in vitro, present in high concentration in the leaves of S. sebiferum, a Chinese folk medicine for shingles.
British division of the international academy of pathology
Authors: Van Den Tweel J , Howat A J , Shepherd N A , Warren B F , Harris C
Published: February 2005

Forthcoming meetings

5–8 July 2005 Joint Meeting with the Pathological Society of Great Britain and Ireland. Newcastle‐Upon‐Tyne, UK

25–26 November 2005 Symposium on Intestinal Pathology. London, UK
Immunophenotypic alteration of the stromal component in minimal deviation adenocarcinoma (‘adenoma malignum’) and endocervical glandular hyperplasia: a study using oestrogen receptor and α‐smooth muscle actin double immunostaining
Authors: Mikami Y , Kiyokawa T , Moriya T , Sasano H
Published: February 2005

Aims : To define the phenotypic alteration of the stromal component in association with destructive invasion which is a crucial feature in distinguishing minimal deviation adenocarcinoma (MDA) from benign endocervical glandular lesions.

Methods and results : We studied endocervical glandular hyperplasias including non‐specific‐type (NEGH) (n = 3) and lobular‐type (LEGH) (n = 8), and minimal deviation adenocarcinoma (MDA) (n = 11), well‐differentiated endocervical adenocarcinoma of usual‐type (WDA) (n = 11), and adenocarcinoma in situ (AIS) (n = 6) of the cervix, by double immunostaining for oestrogen receptor (ER) and α‐smooth muscle actin (α‐SMA) using peroxidase‐ and alkaline phosphatase‐polymer methods, respectively. Glands in NEGH invariably showed nuclear staining for ER, with surrounding ER+/α‐SMA– stromal cells, whereas LEGH also harboured ER+/α‐SMA– spindle cells, but lacked nuclear staining for ER in constituent glands. In contrast, both WDA and MDA displayed accompanying stroma rich in α‐SMA+ spindle cells in close vicinity to the infiltrating neoplastic glands, with only occasional weakly ER+ stromal cells. WDA tended to contain more α‐SMA+ cells. The distribution of α‐SMA+ cells was periglandular (6/11), patchy (6/11), and/or diffuse (4/11) in WDA, whereas in MDA it was periglandular (11/11) and/or patchy (8/11). AIS was surrounded by ER+/α‐SMA− stromal cells. All cases of WDA, MDA, and AIS lacked nuclear staining for ER.

Conclusions : Both MDA and WDA can be distinguished from LEGH and NEGH by identifying surrounding α‐SMA+ stromal cells and the absence or decreased number of ER+ cells, possibly as a result of the desmoplastic reaction with myofibroblasts replacing pre‐existing ER+ stromal cells. In particular, the periglandular distribution of these α‐SMA+ stromal cells can be a clue suggesting destructive stromal invasion in cases of MDA, although occasional glands may lack these cells.
Non‐invasive (intracapsular) carcinoma ex pleomorphic adenoma: recognition of focal carcinoma by HER‐2/neu and MIB1 immunohistochemistry
Authors: Di Palma S , Skálová A , Vanìèek T , Simpson R H W , Stárek I , Leivo I
Published: February 2005

Aims : Non‐invasive carcinoma ex pleomorphic adenoma is defined as a carcinoma arising within the boundaries of a pleomorphic adenoma (PA), but which fails to display invasion beyond the capsule of host PA. Alternative names are intracapsular, in situ, or focal carcinoma. The true nature of non‐invasive carcinoma ex‐PA is still controversial; for example, it is not clear whether it represents early but genuine carcinomatous changes with the genetic make‐up of malignant cells, or simply cytological, possibly metaplastic or ‘bizarre’ changes in PA. Strong overexpression and amplification of HER‐2/neu protein has recently been demonstrated in invasive carcinoma ex‐PA. In addition, data from breast cancer studies suggest that amplification of HER‐2/neu and overexpression of its gene product is mainly involved in the initiation of breast oncogenesis. We sought to establish whether this method could help to demonstrate that what is described as non‐invasive carcinoma ex‐PA is really a genuine malignancy, albeit in an early phase.

Methods and results : Eleven cases of non‐invasive carcinoma (in situ) ex‐PA were studied for HER‐2/neu status using immunohistochemistry and fluorescent in‐situ hybridization (FISH). Cells of focal non‐invasive carcinoma ex‐PA were strongly positive for HER‐2/neu protein, while the cells of the maternal PA were always negative. Two cases of low‐grade non‐invasive myoepithelial carcinoma ex‐PA were negative. In four cases out of a total of six tumours studied by FISH, we detected amplification of HER‐2/neu gene signals in tumour cells of focal, non‐invasive, carcinoma.

Conclusions : The current data suggest that non‐invasive carcinoma ex PA is a genuine carcinoma within a PA. However, the presence of cyto‐nuclear atypia is not sufficient to make a definite diagnosis of malignant change, which requires a combination of morphology and immunohistochemistry.
Overexpression of Ets‐1 proto‐oncogene in latent and clinical prostatic carcinomas
Authors: Alipov G , Nakayama T , Ito M , Kawai K , Naito S , Nakashima M , Niino D , Sekine I
Published: February 2005

Aims : The high incidence of clinically diagnosed prostatic cancer is exceeded by the frequency of tumours detected at autopsy. The Ets‐1 proto‐oncogene is expressed by a variety of malignant and normal tissues. Therefore, in this study, expression of Ets‐1 protein was investigated in ‘latent’ prostatic cancer detected at autopsy, compared with benign prostatic hyperplasia, normal prostatic tissues and clinical prostatic cancer.

Methods and results : Using immunohistochemistry, we analysed Ets‐1 expression in 95 prostatic specimens including 19 cases of latent prostatic carcinoma (LPC) and 55 cases of clinical prostatic carcinoma (CPC), 11 cases of benign prostatic hyperplasia (BPH) and 10 cases of normal prostate (NP). Differences in the incidence of LPC and CPC suggest different courses for the biological progression of prostatic cancer. There was a significant difference in the degree of Ets‐1 expression in CPC and LPC (P < 0.05). Ets‐1 was not expressed in BPH and NP, but in malignant cases (57 of 74; 77.0%) commonly demonstrated immunoreactivity in the tumour cells. In our study the expression of Ets‐1 between benign and malignant, and well, moderately and poorly differentiated adenocarcinomas of prostatic cancer showed significant differences. The presence of Ets‐1 mRNA was confirmed by in‐situ hybridization in human prostatic tissues.

Conclusion : Our results suggest that Ets‐1 might play an important role in carcinogenesis and/or the progression of human prostatic carcinomas.
PDGFRα‐ and c‐kit‐mutated gastrointestinal stromal tumours (GISTs) are characterized by distinctive histological and immunohistochemical features
Authors: Pauls K , Merkelbach‐Bruse S , Thal D , Büttner R , Wardelmann E
Published: February 2005

Aims : To study the immunohistochemical and histological features of 158 gastrointestinal stromal tumours (GISTs), consisting of 137 tumours with mutations in c‐kit and platelet‐derived growth factor receptor‐alpha (PDGFRα) genes and 21 wt‐GISTs. Additionally, we evaluated the localization of PDGFRα in the normal intestine. PDGFRα gene mutations were recently described in a subset of GISTs and it has been hypothesized that PDGFRα‐mutated tumours represent a distinctive entity among GISTs.

Results : PDGFRα was expressed in ganglion bodies of the myenteric plexus and in Schwann cells but not in interstitial cells of Cajal. In contrast to other GISTs, tumours with PDGFRα mutations had an epithelioid phenotype and multinuclear giant cells. Kit was down‐regulated in PDGFRα‐mutated GISTs and PDGFRα expression was decreased in c‐kit mutated tumours. Dot‐like staining of Kit and PDGFRα was associated very frequently with mutation within the respective gene.

Conclusions : Features of PDGFRα‐mutated GISTs are multinuclear giant cells and dot‐like staining for PDGFRα. In contrast, c‐kit‐mutated GISTs display a spindle cell phenotype and Kit‐dots on immunohistochemistry. Our findings not only help to distinguish distinctive entities of GISTs using histological and immunhistochemical features, but also indicate that Kit and PDGFRα are differentially regulated in a subset of GISTs.
Expression of oestrogen receptor and loss of E‐cadherin are diagnostic for gastric metastasis of breast carcinoma
Authors: Van Velthuysen M‐L F , Taal B G , Van Der Hoeven J J M , Peterse J L
Published: February 2005

Aims : To investigate whether immunohistochemical staining for oestrogen receptor (ER)α, progesterone receptor (PgR) and E‐cadherin might be useful to differentiate between metastatic breast carcinoma and primary gastric carcinoma.

Methods : Gastric biopsies of 75 patients containing adenocarcinoma were stained for ERα, PgR and E‐cadherin. Included were: Group A, 28 patients with primary gastric cancer; Group B, 28 patients with an adenocarcinoma containing gastric biopsy and a clinical diagnosis of metastatic breast carcinoma; Group C, all consecutive patients with a positive gastric biopsy in 2001 (n = 19) without clinical history of breast carcinoma and not followed by gastric resection (control group).

Results : All ERα+ or PgR+ carcinomas (n = 20) were of patients with a previous or concurrent history of breast carcinoma: 19 in group B, one in group C. In addition, absence of E‐cadherin staining was seen significantly more often in patients with metastatic breast carcinoma than in patients with primary gastric cancer (P < 0.001).

Conclusion : Positive immunohistochemical staining for ERα or PgR of an adenocarcinoma in a gastric biopsy is diagnostic for metastatic breast carcinoma. Moreover, when carcinoma in a gastric biopsy is negative for E‐cadherin staining, metastatic breast carcinoma should be considered.
Superficial myofibroblastoma of the lower female genital tract: report of a series including tumours with a vulval location
Authors: Ganesan R , McCluggage W G , Hirschowitz L , Rollason T P
Published: February 2005

Aims : To describe the clinical and pathological features of 12 further cases (in 11 patients) of superficial cervicovaginal myofibroblastomas (SCVM), rare tumours that hitherto have been described only in a single series of cases involving the cervix and vagina.

Methods and results : The patients' ages ranged from 23 to 80 years. Ten tumours were located in the vagina and two in the vulva. Three patients had been taking tamoxifen. The tumours ranged in size from 2 to 45 mm and morphologically were well‐circumscribed lesions composed of bland ovoid to spindle‐shaped cells, often with wavy nuclei. These cells were arranged in a variety of architectural patterns and were set in a finely collagenous stroma. Five cases exhibited stromal oedema or myxoid change and in eight cases hyalinized areas with thick, dense collagen bundles were present. Immunohistochemically, there was positivity with vimentin (11 of 11 cases tested), CD34 (six of 12 cases), desmin (nine of 12 cases) and oestrogen receptor (nine of 11 cases). All cases tested were negative for smooth muscle actin, S100, h‐caldesmon, HMB45, and CD31. In this study we expand on the morphological spectrum of these rare lesions and reiterate their association with tamoxifen.

Conculsions : Since these lesions may occur on the vulva, as well as the cervix and vagina, we propose the term ‘superficial myofibroblastoma of the lower female genital tract’.
Vascular endothelial growth factor (VEGF‐C1)‐dependent inflammatory response of podocytes in nephrotic syndrome glomerulopathies in children: an immunohistochemical approach
Authors: Ostalska‐Nowicka D , Zachwieja J , Nowicki M , Kaczmarek E , Siwinska A , Witt M
Published: February 2005

Aims : To analyse expression and distribution of vascular endothelial growth factor (VEGF‐C1), podocalyxin and synaptopodin within renal tissue in nephrotic syndrome glomerulopathies in children.

Methods and results : Renal biopsies performed at the time and in the manner recommended by the World Health Organization. The study group consisted of submicroscopic glomerulonephritis (n = 10), diffuse mesangial proliferation (n = 14) and focal segmental glomerulosclerosis (n = 5). The control tissue consisted of macroscopically normal appearing cortex taken from kidneys resected for localized neoplasms (n = 3). Material for immunohistochemistry was fixed in Bouin's solution and embedded in paraffin. Indirect immunohistochemistry using monoclonal anti‐human antibodies directed against VEGF‐C1, podocalyxin and synaptopodin was employed. The distribution of markers was quantified by computerized image analysis. In non‐sclerosed glomeruli (within podocyte cytoplasm), VEGF‐C1 was more expressed in podocytes of all groups (P < 0.0002), while the distribution of synaptopodin was less expressed in all groups (P < 0.0002). There was no statistical difference between all groups in the expression of podocalyxin.

Conclusions : The increased permeability of the filtration barrier in steroid‐resistant glomerulopathies may be a consequence of subcellular changes in podocytes resulting from decreased expression of synaptopodin. Moreover, impaired permeability of endothelium could be secondary to increased expression of podocyte‐derived VEGF‐C1.
The expression and cellular localization of phosphorylated VEGFR2 in lymphoma and non‐neoplastic lymphadenopathy: an immunohistochemical study
Authors: Pillai G , Cook N , Turley H , Leek R D , Blasquez C , Pezzella F , Harris A L , Gatter K C
Published: February 2005

Aims : To study the expression of phosphorylated vascular endothelial growth factor receptor 2 (VEGFR2), a membrane‐bound tyrosine kinase receptor to vascular endothelial growth factor, in lymphoma and non‐neoplastic lymphadenopathy.

Methods and results : Archival cases (89 cases of lymphoma and 17 cases of non‐neoplastic lymphadenopathy) were studied immunohistochemically with three monoclonal antibodies to the different autophosphorylation sites in the cytoplasmic tail of the receptor. There was increased expression of this receptor in lymphoma and particularly in all cases of peripheral T‐cell lymphoma. In this category, there was nuclear re‐location of this receptor.

Conclusions : This very interesting finding raises the possibility that VEGFR2 may be involved in the transcriptional regulation of this disease. Small molecule inhibitors to this receptor may therefore be a useful adjunct in the therapy of this disease.
A three‐point boundary value problem with an integral condition for a third‐orderpartial differential equation
Authors: Latrous C. , Memou A.
Published: February 2005

We prove the existence and uniqueness of a strong solution for a
linear third‐order equation with integral boundary conditions. The proof uses energy inequalities and the density of the range of the operator generated.
Two Spectrophotometric Assays for Dopamine Derivatives in Pharmaceutical Products and in Biological Samples of Schizophrenic Patients Using Copper Tetramine Complex and Tri‐iodide Reagent
Authors: Nour El-Dien F. A. , Zayed M. A. , Mohamed Gehad. G. , El-Nahas Reham G.
Published: February 2005

Two simple, rapid, and sensitive spectrophotometric methods are proposed for the determination of levodopa (LD). The first method is based on coupling of 4‐aminoantipyrine (4‐AAP) with one of the dopamine derivatives (LD, CD) to give a new ligand that reacts with copper tetramine complex to give intensely colored chelates. The colored products are quantified spectrophotometrically at 525 and 520 nm for LD and CD, respectively. The optimization of the experimental conditions is described. The method has been used for the determination of 19.7–69.0 and 18.1–54.3μg mL−1 of LD and CD, respectively. The accuracy of the method is achieved by the values of recovery (100 ± 0.2%) and the precision is supported by the low standard deviation (SD = 0.17–0.59) and relative standard deviation (CV = 0.4%–1.54%) values. The second method is based on the formation of ion‐pair iodinated inner sphere or outer sphere colored complexes between the LD and triiodide ions at pH 5 and room temperature (23 ± 3°C). This method has been used for the determination of LD within the concentration range 39.44–78.88μg mL−1 with SD = 0.22–0.24 and recovery percent = 100 ± 0.3%. The sensitivity of the two methods is indicated by Sandell′s sensitivity of 0.014–0.019 g cm−2. The results of the two methods are compared with those of the official method. The interference of common drug additives, degradation products, and excipients was also studied. The proposed methods were applied successfully to the determination of the LD‐CD synthetic mixture and Levocare drug. The determination of LD in urine of some schizophrenic patients was applied with good precision and accuracy. The reliability of the methods was established by parallel determinations against the official British pharmacopoeia method.
Deficiency in Cardiac Dystrophin Affects the Abundance of the α‐/β‐Dystroglycan Complex
Authors: Lohan James , Culligan Kevin , Ohlendieck Kay
Published: February 2005

Although Duchenne muscular dystrophy is primarily categorised as a skeletal muscle disease, deficiency in the membrane cytoskeletal protein dystrophin also affects the heart. The central transsarcolemmal linker between the actin membrane cytoskeleton and the extracellular matrix is represented by the dystrophin‐associated dystroglycans. Chemical cross‐linking analysis revealed no significant differences in the dimeric status of the α‐/β‐dystroglycan subcomplex in the dystrophic mdx heart as compared to normal cardiac tissue. In analogy to skeletal muscle fibres, heart muscle also exhibited a greatly reduced abundance of both dystroglycans in dystrophin‐deficient cells. Immunoblotting demonstrated that the degree of reduction in α‐dystroglycan is more pronounced in matured mdx skeletal muscle as contrasted to the mdx heart. The fact that the deficiency in dystrophin triggers a similar pathobiochemical response in both types of muscle suggests that the cardiomyopathic complications observed in x‐linked muscular dystrophy might be initiated by the loss of the dystrophin‐associated surface glycoprotein complex.
Artificial Cell Therapy: New Strategies for the Therapeutic Delivery of Live Bacteria
Authors: Prakash Satya , Jones Mitchell Lawrence
Published: February 2005

There has been rapid growth in research regarding the use of live bacterial cells for therapeutic purposes. The recognition that these cells can be genetically engineered to synthesize products that have therapeutic potential has generated considerable interest and excitement among clinicians and health professionals. It is expected that a wide range of disease modifying substrates such as enzymes, hormones, antibodies, vaccines, and other genetic products will be used successfully and will impact upon health care substantially. However, a major limitation in the use of these bacterial cells is the complexity of delivering them to the correct target tissues. Oral delivery of live cells, lyophilized cells, and immobilized cells has been attempted but with limited success. Primarily, this is because bacterial cells are incapable of surviving passage through the gastrointestinal tract. In many occasions, when given orally, these cells have been found to provoke immunogenic responses that are undesirable. Recent studies show that these problems can be overcome by delivering live bacterial cells, such as genetically engineered cells, using artificial cell microcapsules. This review summarizes recent advances in the therapeutic use of live bacterial cells for therapy, discusses the principles of using artificial cells for the oral delivery of bacterial cells, outlines methods for preparing suitable artificial cells for this purpose, addresses potentials and limitations for their application in therapy, and provides insight for the future direction of this emergent and highly prospective technology.