bluuebunny/crossref_metadata_2025 huggingface dataset:
Avidin attachment to biotinylated amino groups of the erythrocyte membrane eliminates homologous restriction of both classical and alternative pathways of the complement
Authors: Muzykantov Vladimir R. , Smirnov Mikhail D. , Klibanov Alexander L.
Published: July 2002

Lysis of avidin‐coated biotinylated sheep red blood cells (RBC) via the classical pathway of homologous (sheep) and heterologous (guinea pig) complement has been studied. The minimal surface density of avidin inducing antibody‐dependent lysis via the classical pathway is smaller than that inducing antibody‐independent lysis via the alternative pathway. Heterologous lysis via the classical pathway does not depend on the mode of avidin attachment: both biotinylation of membrane amino groups and insertion of biotinyl‐lipid into the membrane provide the same lysis of avidin‐coated RBCs by guinea pig serum in the presence of anti‐avidin antibody. Avidin‐free sheep RBC sensitized with hemolytic anti‐RBC antibody were lysed by guinea pig, but not by sheep serum, confirming high efficiency of homologous restriction of the complement. However, avidin‐coated RBCs were lysed by homologous serum in the presence of anti‐avidin antibody at low surface density of avidin attached. The elimination of the homologous restriction depends on the mode of avidin attachment: biotinylation of membrane amino groups provides antibodymediated lysis via the classical pathway of homologous complement, while insertion of biotinyl‐lipid does not provide lysis.
Production and characterization of monoclonal antibodies specific for different epitopes of human tenascin
Authors: Balza Enrica , Siri Annalisa , Ponassi Marco , Caocci Fabio , Linnala Auli , Virtanen Ismo , Zardi Luciano
Published: July 2002

We have obtained and characterized 11 monoclonal antibodies (mAbs) specific for different domains of human tenascin (TN). Five of these mAbs reacted with epitopes contained in the TN area that undergoes alternative splicing and are thus able to recognize specific TN isoforms. These mAbs are a useful tool to study the expression and distribution of TN and its different isoforms in normal and pathological tissues.
Impaired gene transcription and nuclear protein kinase C activation in the brain and liver of aged rats
Authors: Rogue Patrick J. , Ritz Marie-Françoise , Malviya Anant N.
Published: July 2002

The expression of the hsp70 and c‐fos genes and the activation of nuclear protein kinase C (PKC) were studied in young and aged whole rats under heat‐shock conditions. The induction of hsp70 and c‐fos genes by heat shock were decreased several fold in the brain as well as in the liver of senescent animals. Nuclear run‐off transcription assay indicated that this age‐related impairment could be attributed to a block at the level of transcription. Nuclear PKC activation by heat shock was not apparent in old animals. Nuclear PKC involvement in the repression of transcription during senescence is postulated.
Possible regulation of CFTR‐chloride channels by membrane‐bound phosphatases in pancreatic duct cells
Authors: Becq F. , Fanjul M. , Merten M. , Figarella C. , Hollande E. , Gola M.
Published: July 2002

We have studied CFTR‐Cl− channels in non‐CF CAPAN‐1 and in CFTR‐transfected CFPAC‐PLJ‐CFTR‐6 epithelial cells from human pancreas. Theophylline and IBMX induced the opening of cell‐attached CFTR‐Cl− channels. Theophylline, IBMX and the alkaline phosphatase (AP) inhibitor levamisole enhanced the activity of excised channels and reduced by 70–75% the apical membrane‐associated APs activity. Okadaic acid had no effect on APs and channel activities. A polyclonal anti‐alkaline phosphatase antibody (which detected apical APs) reduced APs activity and activated quiescent excised chloride channels. These results suggest that CFTR channels may be regulated by membrane‐bound phosphatases.
The tyrosine kinase inhibitors methyl 2,5‐dihydroxycinnamate and genistein reduce thrombin‐evoked tyrosine phosphorylation and Ca2+ entry in human platelets
Authors: Sargeant Paul , Farndale Richard W. , Sage Stewart O.
Published: July 2002

Platelet activation is associated with the phosphorylation of a number of platelet proteins at tyrosine residues. The significance of this is unknown. Here we have investigated the effects of two tyrosine kinase inhibitors, methyl 2,5‐dihydroxycinnamate and genistein, on thrombin‐evoked protein tyrosine phosphorylation and Ca2+ signal generation in fura‐2‐loaded human platelets. Both compounds inhibited thrombin‐evoked tyrosine phosphorylation and reduced the elevation of [Ca2+], in the presence, but not the absence, of external Ca2+. This suggested a selective inhibition of thrombin‐evoked Ca2+ entry but not release from internal stores. Both compounds also reduced thrombin‐evoked Mn2+ entry. In contrast, selective blockade of protein kinase C with Ro 31/8220‐002 potentiated the thrombin‐evoked Ca2+ signal. These data are compatible with a role for protein tyrosine phosphorylation contributing to thrombin‐evoked Ca2+ entry in human platelets.
d‐α‐Tocopherol inhibits low density lipoprotein induced proliferation and protein kinase C activity in vascular smooth muscle cells
Authors: Özer N.K. , Palozza P. , Boscoboinik D. , Azzi A.
Published: July 2002

Native and malondialdehyde modified low density lipoproteins have been shown to stimulate smooth muscle cell proliferation (A7r5) in vitro. The stimulation is associated with an increase of protein kinase C activity. d‐α‐Tocopherol, at physiological concentrations, has been found to inhibit both protein kinase C activity and cell proliferation.
Nonendothelial‐derived nitric oxide activates the ATP‐sensitive K+ channel of vascular smooth muscle cells
Authors: Miyoshi Hirokazu , Nakaya Yutaka , Moritoki Hideki
Published: July 2002

To determine whether endogenous nitric oxide (NO) opens the ATP‐sensitive K+ channel (KATP channel), we investigated the effect of nonendothelial‐derived NO on this channel in cultured smooth muscle cells of the porcine coronary artery by the patch‐clamp technique. In the cells pretreated with endotoxin, the addition of 10−4 M l‐arginine generated NO and activated the KATP channel. Activation of this channel was suppressed by pretreatment with 10−3 M N
G‐methyl‐l‐arginine or 10−3 M N
x‐nitro‐l‐arginine methyl ester, each of which is a specific antagonist of the l‐arginine‐NO pathway, and by 10−6 M Methylene blue, which blocks guanylate cyclase. The activation of the KATP channel by l‐arginine‐NO pathway is expected to produce hyperpolarization of the cell membrane and relaxation of vascular smooth muscle cells.
Molecular cloning of a novel protein‐tyrosine phosphatase containing a membrane‐binding domain and GLGF repeats
Authors: Maekawa Kazuhiko , Imagawa Noriko , Nagamatsu Masaaki , Harada Shigenori
Published: July 2002

A full‐length cDNA encoding a novel cytosolic protein‐tyrosine phosphatase (PTP), PTP‐BAS, was cloned from human basophils. Due to in‐frame deletions in the coding region, PTP‐BAS exists in three isoforms: 7,455 bp (2,485 aa) for type 1, 7,398 bp (2,466 aa) for type 2 and 6,882 bp (2,294 aa) for type 3. All three isoforms contain a single PTP catalytic domain at the carboxyl termini as well as two distinct structural sequences. Amino terminal sequences of 300 amino acids are homologous to membrane‐binding domains of eytoskeleton‐associated proteins. Three 90 amino acid internal repetitive sequences are homologous to the GLGF repeats found in guanylate kinase proteins. PTP‐BAS was expressed in various human tissues, especially highly in the kidney and lung. Interestingly, the BAS mRNA level in the fetal brain was remarkably high.
cis‐Acting signals and trans‐acting proteins are involved in tau mRNA targeting into neurites of differentiating neuronal cells
Authors: Behar L. , Marx R. , Sadot E. , Barg J. , Ginzburg I.
Published: July 2002

Tau microtubule‐associated protein is a neuron specific protein found primarily in axons and is developmentally regulated. The function of tau is in stabilization of microtubules, which is important in establishing and maintaining neuronal morphology. Axonal localization of tau involves a multistep process which is studied in differentiating primary neuronal culture. The initial step involves sorting and subcellular localization of its encoding mRNA into the proximal portion of the axon. Using the transfection assay into neuronal cells, we have demonstrated that sequences located in the 3′‐untranslated region include a cis‐acting signal which is involved in tau mRNA targeting. In addition, using ultraviolet cross‐linking assay, two RNA‐binding proteins of 43 and 38 kDa were identified, that exhibit specific binding to a minimal sequence of 91 nucleotides located within the same functional region, which is involved in targeting. The 43 and 38‐kDa RNA‐binding proteins are present in cytoplasmic extracts, prepared from neuronal cells, and in isolated microtubule preparations. Our results support a novel model in which cis‐acting signals, together with RNA‐binding proteins, are involved in the targeting of tau mRNA, that may ultimately lead to its axonal localization.
Cholinergic properties of neurons differentiated from an embryonal carcinoma cell‐line (P19)
Authors: Parnas D. , Linial M.
Published: July 2002

P19 is a mouse‐derived embryonal carcinoma cell‐line capable of differentiation toward ectodermal, mesodermal and endodermal lineages. Following treatment with retinoic acid these cells differentiated into neurons, astrocytes and fibroblast‐like cells.

We induced P19 differentiation under conditions which lead to a homogeneous neuronal culture (> 95% neurons). Under these conditions, most cells (∼85%) express high levels of the cholinergic markers acetyl cholinesterase and choline acetyltransferase while ∼10% of cells express the GABAergic marker glutamic acid decarboxylase. While the proportion of the GABAergic neurons is constant at different culture conditions, the cholinergic phenotype is suppressed at high cell densities. The cholinergic nature of P19 neurons is also evident in their ability to form contacts with a muscle cell‐line — C2. At day 10 of differentiation cells are capable of depolarization‐dependent acetylcholine release. The release is Ca2+ dependent, and drops to baseline levels at 0.5 mM Ca2+. The cells also respond to sub‐nM levels of α‐latrotoxin by acetylcholine release. All major proteins implicated in synapse functionality are expressed prior to day 10 at both at RNA and protein levels. However, the expression pattern of each gene is unique. The genes include cytoskeletal proteins, synaptic vesicle proteins and terminal specific proteins.

We suggest that this cell‐line can serve as an in‐vitro model system for the study of neuronal phenotype acquisition. Under our conditions, the P19 cells can also provide a system in which to study the differentiation of functional cholinergic neurons.
Human microglia cultures: A powerful model to study their origin and immunoreactive capacity
Authors: Lauro G.M. , Babiloni D. , Buttarelli F.R. , Starace G. , Cocchia D. , Ennas M.G. , Sogos V. , Gremo F.
Published: July 2002

In this paper, we report that pure cultures of human microglia were obtained from long‐term astrocytic cultures of human fetal brain. After five to six months and repeated cell passages, macrophage‐like cells started to spontaneously form in vitro, so that in two to three weeks the whole culture was populated by them. These cells were grown up to over 50 passages in culture and analyzed for morphology, specific marker positivity, growth rate and major histocompatibility complex (MHC) antigen expression with or without gamma‐interferon (IFN) stimulation. We found that, regardless of embryonic age of original cultures (10–15 weeks of gestation), cultures showed a remarkable homogeneity and purity and over 90 stained for typical microglial markers. Under basal conditions, two cell subpopulations similar to those described in vivo, we observed: the reactive ‘ameboid’ type and the resting ‘ramified’ one, the latter increasing with time in vitro and cell passages. Both cell subpopulations were capable of active phagocytosis and of high‐rate proliferation. They spontaneously expressed low levels of MHC class II antigens, but were negative for MHC class I. Stimulation with gamma‐interferon lymphokine upregulated the MHC class II expression as well as the MHC class I heavy chain form in ameboid, ‘reactive’ cells but not in the ramified ones. We also found that β2 microglobulin, already expressed in basal conditions, was dissociated from HLA A‐B‐C molecules in lymphokine‐stimulated cells at early passages. The physiological significance of these data, as well as the possible correlation with in vivo ontogenetic modifications, are also discussed.
Recognition and treatment of patients with sleep problems in general health care
Authors: Lecrubier Y , Weiller E , Privett M , Boyer P , Maier W , Üstün TB , Sartorius N ,   Principal investigators of the WHO International Study on Psychological Problems in General Health Care
Published: July 2002

The form, frequency and burden of sleep problems in general health care in the WHO Collaborative Study on Psychological Problems are presented in this same issue. This study was conducted in 5,438 primary care patients belonging to 15 centres in 14 countries after 25,916 patients were screened with the GHQ-12. Diagnoses according to the International Classification of Diseases (ICD-10) were obtained from the Composite International Diagnostic Interview (CIDI-PHC). The presence of difficulties falling asleep, staying asleep, waking too early or sleeping too much were assessed by the CIDI. General practitioners (GPs) gave their opinion on the existence of a psychological problem and/or a physical disease and indicated what therapeutic intervention was proposed to patients they recognized as psychological cases. The existence of sleep problems increased the recognition of patients with psychiatric disorders by their GP. No specific subtype of sleep problems influences recognition. The existence of a somatic disease had little influence on the identification of sleep problems as psychological cases. Sleep problems were not frequently expressed as a main presenting complaint. In such a case the recognition rate of patients with ICD-10 diagnosis was unchanged but a sedative treatment was offered more frequently. Overall, 85.9% of patients with sleep problems and recognized as cases were offered treatment, 53.9% drug treatment. The most frequent treatment consumed was stimulants, tonics or vitamins while the most frequently prescribed were antidepressants, hypnotic and anxiolytics with rather similar proportions. Drug consumption was substantially lower than drug prescription.
A psychometric evaluation of dementia rating scales
Authors: Kørner A , Lauritzen L , Bech P
Published: July 2002

The paper discusses the relevance of sufficient psychometric standards for dementia rating scales. The concurrent, convergent and construct validity of the Mini Mental State Examination (MMSE), the Alzheimer's Disease Assessment Scale (ADAS) and the CAMCOG are assessed. The Clinical Global Impressions and the Global Deterioration Scale are used as global scales. The concurrent and convergent validity are satisfactory. The construct validity expressed by the Cronbach and Loevinger coefficient are very good for all scales and subscales. The Mokken's single item coefficients show that the MMSE has the best individual hierarchical fit, the item reading can be left out. The ADAS is less uni-dimensional, eight items can be left out. The CAMCOG consists of too many items to apply the Mokken's single item coefficients or the Loevinger coefficient. Instead, the CAMCOG subscales are analyzed. This results in a possible reduction of the CAMCOG by 30 items to a total of 35 items. The factor analysis reveals two factors in both the MMSE and the ADAS while the number of observations does not allow a factor analysis of the CAMCOG to be performed.
Triple helix assembly and processing of human collagen produced in transgenic tobacco plants
Authors: Ruggiero F , Exposito J.-Y , Bournat P , Gruber V , Perret S , Comte J , Olagnier B , Garrone R , Theisen M
Published: July 2002

The use of tobacco plants as a novel expression system for the production of human homotrimeric collagen I is presented in this report. Constructs were engineered from cDNA encoding the human proα1(I) chain to generate transgenic tobacco plants expressing collagen I. The recombinant proα1(I) chains were expressed as disulfide‐bonded trimers and were shown to fold into a stable homotrimeric triple helix. Moreover, the recombinant procollagen was subsequently processed to collagen as it occurs in animals. Large amounts of recombinant collagen were purified from field grown plant material. The data suggest that plants are a valuable alternative for the recombinant production of collagen for various medical and scientific purposes.
Mitochondria: execution central
Authors: Gottlieb Roberta A.
Published: July 2002

Mitochondria play an essential function in eukaryotic life and death. They also play a central role in apoptosis regulation, reflected by the convergence of Bcl‐2 family members on the mitochondrial outer membrane, and the presence of ‘death factors’ in the intermembrane space. Mitochondrial structure and function must be taken into consideration when evaluating mechanisms for cytochrome c release. The core machinery for caspase activation is conserved from Caenorhabditis elegans to man, and we consider parallels in the role of mitochondria in this process.
P2 purinergic receptors of human eosinophils: characterization and coupling to oxygen radical production
Authors: Ferrari Davide , Idzko Marco , Dichmann Stefan , Purlis Daniela , Virchow Christian , Norgauer Johannes , Chiozzi Paola , Di Virgilio Francesco , Luttmann Werner
Published: July 2002

Extracellular nucleotides elicit multiple responses in eosinophils but no information on expression of purinergic receptors in these cells is available so far. In the present study we show that human eosinophils express the following P2Y and P2X subtypes: P2Y1, P2Y2, P2Y4, P2Y6, P2Y11, and P2X1, P2X4, P2X7, whose stimulation results in intracellular Ca2+ increase and production of large amounts of reactive oxygen intermediates. These events are stimulated or inhibited, respectively, by P2 receptor agonists or antagonists.
Functional distinction of human EAG1 and EAG2 potassium channels
Authors: Schönherr Roland , Gessner Guido , Löber Karsten , Heinemann Stefan H.
Published: July 2002

Human ether à go‐go potassium channel 2 (hEAG2) was cloned and its properties were compared with the previously characterized isoform hEAG1. In the Xenopus oocyte expression system the time course of activation was about four times slower and the voltage required for half‐maximal subunit activation was about 10 mV greater for hEAG2 channels. However, its voltage dependence was smaller and, therefore, hEAG2 channels start to open at more negative voltages than hEAG1. Coexpression of both isoforms and kinetic analysis of the resulting currents indicated that they can form heteromeric channel complexes in which the slow activation phenotype of hEAG2 is dominant. Upon expression in mammalian cells, quinidine blocked hEAG1 channels (IC50 1.4 μM) more potently than hEAG2 channels (IC50 152 μM), thus providing a useful tool for the functional distinction between hEAG1 and hEAG2 potassium channels.
Construction and characterization of the soybean leaf metalloproteinase cDNA1
Authors: Pak Jhang Ho , Liu Chuan Y , Huangpu Jun , Graham John S
Published: July 2002

© 1997 Federation of European Biochemical Societies.
Evidence for light‐dependent and light‐independent protein dephosphorylation in chloroplasts
Authors: Elich Tedd D , Edelman Marvin , Mattoo Autar K
Published: July 2002

A number of photosystem II (PSII) associated proteins, including core proteins D1, D2 and CP43, and several proteins of the LHCII complex, are phosphorylated by a thylakoid‐bound, redox‐regulated kinase(s). We demonstrate here that the compound propyl gallate is an effective inhibitor of LHCII phosphorylation in vivo while having little effect on PSII core protein phosphorylation. Using this inhibitor, we demonstrate that LHCII dephosphorylation is insensitive to light in vivo. Taken together with our previous conclusion (Elich et al., EMBO J. 12 (1993) 4857–4862) that PSII core protein dephosphorylation is light‐stimulated, our data suggest the presence of multiple phosphatases responsible for thylakoid protein dephosphorylation in vivo.
Nitric oxide synthase isoform III gene expression in rat liver is up‐regulated by lipopolysaccharide and lipoteichoic acid
Authors: Bucher M , Ittner K.P , Zimmermann M , Wolf K , Hobbhahn J , Kurtz A
Published: July 2002

This study was done to investigate the influence of Gram‐negative and Gram‐positive sepsis on the expression of the three isoforms of nitric oxide synthase (NOS) gene in rat liver and kidney. Male Sprague‐Dawley rats were treated with lipopolysaccharide (LPS, 10 mg/kg i.v.) as an in vivo model for Gram‐negative sepsis or lipoteichoic acid (LTA, 10 mg/kg i.v.) as an in vivo model for Gram‐positive sepsis. Animals were killed 12 h and 24 h after i.v. treatment. NOS mRNA of the three isoforms was determined by RNase protection assay. NOS II gene expression was strongly induced after LPS or LTA treatment in rat liver and kidney, indicating the efficacy of this treatment to induce sepsis. We found no change of NOS I gene expression after LPS or LTA injection in rat liver and kidney. NOS III gene expression was increased about 8‐fold 12 h and about 5‐fold 24 h after induction of sepsis in the rat liver whereas in the kidney there was no significant increase in NOS III gene expression. After correction for length NOS III mRNA was about 4‐ and 40‐fold more abundant 12 h and 24 h after LPS treatment than NOS II mRNA in the liver, respectively. Twelve and 24 h after LTA treatment NOS III mRNA was about 18‐ and 140‐fold more abundant than NOS II in the liver. These findings suggest that NOS III is an even more potent source of NO than NOS II in the liver after stimulation with LPS or LTA.
Cloning and characterization of subtracted cDNAs from a human ciliary body library encoding TIGR, a protein involved in juvenile open angle glaucoma with homology to myosin and olfactomedin
Authors: Ortego Javier , Escribano Julio , Coca-Prados Miguel
Published: July 2002

A group of cDNAs isolated from a subtractive ciliary body library of a normal human eye donor revealed 100% identity with TIGR a candidate gene responsible for juvenile open angle glaucoma [Science 275 (1997) 668–670]. Several structural features of the deduced human protein have been noted: a cleavable N‐terminal signal peptide, a periodic repetition at the N‐terminus of leucine and arginine residues at every seventh and eleven position respectively in helix conformation (leucine zipper‐like motif) exhibiting homology with myosin, and with olfactomedin in the C‐terminus. The mRNA for TIGR is abundantly expressed in the ciliary body, iris, heart and skeletal muscle.
Specific reg II gene overexpression in the non‐obese diabetic mouse pancreas during active diabetogenesis
Authors: Baeza N , Sanchez D , Vialettes B , Figarella C
Published: July 2002

The reg gene, previously described in islets of 90% pancreatectomized and nicotinamide‐treated rats, has been shown to be expressed in many pharmacological or surgical animal models of beta cell regeneration. We have studied the non‐obese diabetic (NOD) mouse, which represents a good model of spontaneous autoimmune diabetes in which regenerative processes have recently been demonstrated. Two reg genes have been described in the mouse genome, both recognized by the human reg cDNA. In a previous work, using the human probe, we have demonstrated a strong correlation between pancreatic reg gene expression and the likelihood of developing diabetes. In the present study, we have examined the respective levels of both mouse reg I and reg II mRNA in the NOD mouse pancreas using their specific cDNA probes. We found that reg II expression was specifically prevalent compared to reg I, irrespective of sex or state of the disease. Reg II mRNA was particularly increased in overtly diabetic female mice and in cyclophosphamide‐treated male mice. These data underline the need to study separately the reg genes using specific probes and show that both reg genes are subjected to various regulations, strongly suggesting that their physiological functions may be different.
A novel lipopolysaccharide inducible C‐C chemokine receptor related gene in murine macrophages
Authors: Shimada Takahiro , Matsumoto Makoto , Tatsumi Yoichi , Kanamaru Akihisa , Akira Shizuo
Published: July 2002

To identify genes induced in activated macrophages, we screened a cDNA library prepared from the lipopolysaccharide (LPS)‐treated cell line, RAW264, using the suppression subtractive hybridization technique. One of the clones isolated was dramatically induced by LPS in macrophages. The predicted protein sequence of this gene contains the domain unique to seven transmembrane receptors, and shows similarity with mouse C‐C chemokine receptor 5 (CCR5). Therefore, we designated it LPS inducible C‐C chemokine receptor related gene (L‐CCR). Northern blot analysis revealed that L‐CCR was specifically expressed in differentiated macrophages after LPS stimulation. These results show that L‐CCR is a novel C‐C chemokine receptor related gene induced by LPS in macrophages and may play an important role in inflammatory responses.
Bcl‐2 and Bax are involved in the sympathetic protection of brown adipocytes from obesity‐linked apoptosis
Authors: Briscini Luca , Tonello Cristina , Dioni Laura , Carruba Michele O. , Nisoli Enzo
Published: July 2002

Obesity is linked to functional brown adipose tissue (BAT) atrophy, partially due to adipocyte apoptosis. The brown adipocytes of obese rats have lower Bcl‐2/Bax mRNA and protein ratios than those of their lean littermates. Exposure to a low temperature for three days markedly increased the Bcl‐2/Bax ratio, by increasing the noradrenergic output to BAT, which has previously been shown to reduce apoptotic cell death. This effect could be mimicked in vitro by the addition of noradrenaline (NA) to brown adipocytes differentiated in culture. Micromolar NA concentrations increased the Bcl‐2/Bax mRNA and protein ratios, and protected against serum deprivation‐induced apoptosis. We conclude that NA acts by modulating bcl‐2 and bax gene expression.
CD95‐mediated apoptosis: no variation in cellular sensitivity during cell cycle progression
Authors: Hueber Arno , Durka Silke , Weller Michael
Published: July 2002

Sensitivity of CD95‐mediated apoptosis has been reported to vary during cell cycle progression (FEBS Lett. (1997) 412, 91–93). Here, we report that three human glioma cell lines with different p53 status (i) undergo growth arrest and synchronous cell cycle re‐entry after prolonged serum deprivation, (ii) do not exhibit cell cycle‐related changes in CD95 expression at the cell surface, and (iii) do not exhibit cell cycle‐related changes in susceptibility to DC95 ligand‐induced apoptosis. In contrast, cell cycle‐specific activity was demonstrated for various cancer chemotherapy drugs. Further, CD95 expression and susceptibility to CD95 ligand‐induced apoptosis does not vary during cell cycle progression of Jurkat T cells, HeLa cervical carcinoma and HepG2 hepatocellular carcinoma cells. These results do not support a role for the cell cycle phase as an important predictor of vulnerability to CD95‐mediated apoptosis.
Identification and characterization of a 14 kDa human protein as a novel parvulin‐like peptidyl prolyl cis/trans isomerase
Authors: Uchida Takafumi , Fujimori Fumihiro , Tradler Thomas , Fischer Gunter , Rahfeld Jens-U
Published: July 2002

A second member of the parvulin family of peptidyl‐prolyl cis/trans isomerases was identified in a human lung cDNA library. The gene encoded a protein named hPar14 that has 131 amino acid residues and a molecular mass of 13 676 Da. Sequence comparison showed 34.5% identity to E. coli Par10 and 34% identity to human Pin1 (hPar18) within a C‐terminal region of 87 or 120 amino acid residues, respectively. In comparison to the E. coli Par10, hPar14 possesses a N‐terminal extension of 41 amino acid residues. This extension does not contain a polyproline II helix‐binding motif typical of the known eukaryotic parvulins. The hPar14 does not accelerate the cis to trans interconversion of oligopeptides with side chain‐phosphorylated Ser(Thr)‐Pro moieties as hPin1 did. In contrast, it showed preference of an arginine residue adjacent N‐terminal to proline. Northern blot analysis revealed expression of the gene within various human tissues like heart, placenta, liver, kidney and pancreas.
Genes induced by growth arrest in a pancreatic β cell line: identification by analysis of cDNA arrays
Authors: Zimmer Yitzhak , Milo-Landesman Dalit , Svetlanov Anton , Efrat Shimon
Published: July 2002

Pancreatic β cell lines are a potentially attractive source of material for cell therapy of insulin‐dependent diabetes mellitus. However, induction of proliferation in post‐mitotic, differentiated β cells is likely to affect the expression of multiple genes associated with cell function, resulting in dedifferentiation. We have developed a murine β cell line by conditional transformation with the SV40 T antigen oncoprotein. These cells can undergo reversible induction of proliferation and growth arrest. Here we utilized this model to identify differences in gene expression between proliferating and quiescent β cells, by analyzing known β cell genes and differentially secreted proteins, as well as by a systematic survey of a mouse cDNA array. Our findings demonstrate that growth arrest stimulates expression of the insulin gene and genes encoding components of the insulin secretory vesicles. Screening of the cDNA array revealed the activation of multiple genes following growth arrest, many of them novel genes which may be related to β cell function. Characterization of these genes is likely to contribute to our understanding of β cell function and the ability to employ β cell lines in cell therapy of diabetes.
Detergent‐solubilized Escherichia coli cytochrome bo3 ubiquinol oxidase: a monomeric, not a dimeric complex
Authors: Musatov Andrej , Ortega-Lopez Jaime , Demeler Borries , Osborne Jeffrey P , Gennis Robert B , Robinson Neal C
Published: July 2002

The protein molecular weight, M
r, and hydrodynamic radius, R
s, of Triton X‐100‐solubilized Escherichia coli cytochrome bo
3 were evaluated by computer fitting of sedimentation velocity data with finite element solutions to the Lamm equation. Detergent‐solubilized cytochrome bo
3 sediments as a homogeneous species with an s
20,w of 6.75 s and a D
20,w of 3.71×10−7 cm2/s, corresponding to a R
s of 5.8 nm and a M
r of 144 000±3500. The protein molecular weight agrees very well with the value of 143 929 calculated from the four known subunit sequences and the value of 143 025 measured by MALDI mass spectrometry for the histidine‐tagged enzyme. We conclude that detergent‐solubilized E. coli ubiquinol oxidase is a monomeric complex of the four known subunits.
Distribution and quantification of immunoreactive orexin A in rat tissues
Authors: Taheri Shahrad , Mahmoodi Mehdi , Opacka-Juffry Jolanta , Ghatei Mohammad A. , Bloom Stephen R.
Published: July 2002

A sensitive and specific radioimmunoassay for orexin A was developed. Orexin A immunoreactivity was found to be confined to the central nervous system (CNS) with the highest concentrations in the hypothalamus, inferior and superior colliculi and brainstem. Within the hypothalamus, the highest levels were found in the lateral and posterior hypothalamus. These regions had a greater orexin A content in females compared to males. The orexin A content of hypothalamic regions did not change with fasting and no difference was noted in hypothalami of rats fed a high fat diet. The hypothalamic orexin A content was not different in obese Zucker rats compared to lean controls. Thus, orexin A has a wide distribution in the CNS, but appetite regulation may not be its main function.
Function of the second nucleotide‐binding fold in the CFTR chloride channel
Authors: Zerhusen Bryan , Ma Jianjie
Published: July 2002

To test the role of nucleotide‐binding fold (NBF) 2 and its interaction with the regulatory (R) domain in the function of the cystic fibrosis transmembrane conductance regulator (CFTR) channel, we used three deletion mutants of CFTR: ΔR(708‐835), ΔNBF2(1185‐1349) and ΔR‐ΔNBF2. In lipid bilayers, ΔNBF2 channel activity is ATP‐ and cAMP‐dependent protein kinase (PKA)‐dependent, but unlike wild‐type (wt) CFTR, it displays a reduced activity and insensitivity to 5′‐adenylylimidodiphosphate (AMP‐PNP). Both ΔR and ΔR‐ΔNBF2 channels are PKA‐independent, but ΔR activity is reduced whereas ΔR‐ΔNBF2 activity is increased. Deletion of NBF2 from CFTR affects protein trafficking and channel gating kinetics. The data suggest that NBF2 could have inhibitory and stimulatory roles in CFTR activity by interaction with NBF1 directly or indirectly via the R domain.
Determination of cDNA, gene structure and chromosomal localization of the novel human 17β‐hydroxysteroid dehydrogenase type 7
Authors: Krazeisen Antje , Breitling Rainer , Imai Kenji , Fritz Stephanie , Möller Gabriele , Adamski Jerzy
Published: July 2002

We have identified human 17β‐hydroxysteroid dehydrogenase type 7 (17β‐HSD 7). The novel human cDNA encodes a 37 kDa protein that shows 78 and 74% amino acid identity with rat and mouse 17β‐HSD 7, respectively. These enzymes are responsible for estradiol production in the corpus luteum during pregnancy, but are also present in placenta and several steroid target tissues (breast, testis and prostate) as revealed by RT‐PCR. The human 17β‐HSD 7 gene (HSD17B7) consists of nine exons and eight introns, spanning 21.8 kb and maps to chromosome 10p11.2 close to susceptibility loci for tumor progression, obesity and diabetes. The HSD17B7 promoter (1.2 kb) reveals binding sites for brain‐specific and lymphoid transcription factors corresponding to additional expression domains in hematopoietic tissues and the developing brain as identified by in silico Northern blot.
The risk of urinary incontinence of parous women who delivered only by cesarean section
Authors: Faúndes A , Guarisi T , Pinto‐Neto A.M
Published: July 2002

Objectives: To evaluate the association of route of delivery with prevalence of urinary incontinence. Methods: Women with menopause who consulted at the Department of Gynecology and Obstetrics, Universidade Estadual de Campinas, were interviewed and examined to detect urinary incontinence. Obstetrics history and other possible factors that could influence incontinence were also recorded. Results: Urinary incontinence was diagnosed in 98 women (cases) while 91 were free of this problem. The risk of urinary incontinence was approximately five times higher among women with one or more pregnancies than among nulligravida, and 3.5 times higher among women who had had only cesarean sections than among nullipara. Conclusions: Women cannot prevent urinary incontinence by delivering exclusively by cesarean section.
Influence of compression of the inferior vena cava in the late second trimester on uterine and umbilical artery blood flow
Authors: Ryo E. , Okai T. , Kozuma S. , Kobayashi K. , Kikuchi A. , Taketani Y.
Published: July 2002

Objective: To evaluate the inferior vena cava compression and its influence on the uterine and umbilical artery blood flow in the late second trimester when the mother is supine. Methods: The inferior vena cava diameter was measured by ultrasound B mode scan, and Doppler flow velocimetry of the uterine and umbilical artery was performed in 90 women at 24–27 weeks in the supine and complete left lateral position. Results: The inferior vena cava diameter in the supine position was significantly smaller than that in the lateral position. The degree of the vena cava compression was associated with an elevation in the uterine artery RI in the supine position. The umbilical artery RI did not couple with either the degree of the compression or the changes in the uterine artery RI. Conclusion: The inferior vena cava is compressed in the majority of pregnant women in the second trimester, and the compression may affect the uterine artery blood flow but not the fetal circulation.
Radiation dose to patients from radiopharmaceuticals: (Addendum 2 to ICRP Publication 53) ICRP Publication 80 Approved by the Commission in September 1997
Authors: Valentin J.
Published: July 2002

A joint Task Group of ICRP Committees 2 (Doses from Radiation Exposures) and 3 (Protection in Medicine; lead Committee for this report) has prepared a compilation of data on radiation dose to patients from radiopharmaceuticals.

The report provides biokinetic models, absorbed doses, and effective doses, using ICRP Publication 60 dosimetry, for 10 new radiopharmaceuticals: [Methyl-11C]thymidine; [2-11C]thymidine; 14C urea (incl. carbon dioxide and bicarbonate); 15O water; 99mTc HIG, Pertechnegas, Technegas, and tetrofosmin; and 111In HIG and octreotide.

It also provides recalculated dose data for the 19 most frequently used radiopharmaceuticals from ICRP Publication 53, using ICRP Publication 60 dosimetry, viz. 18F FDG; 51Cr EDTA; 67Ga citrate; 75Se SeHCAT; 99mTc DMSA, DTPA, RBC, IDA, large colloids, WBC, MAA, non-absorbable markers, pertechnetate, and phosphates and phosphonates; 123I Hippuran and MIBG; 131I Hippuran and NP59; and 201Tl thallous ion. Printing errors detected in ICRP Publication 53 are also listed.

Furthermore, the report reproduces with minor corrections and updates, and therefore supersedes, the information on 6 radiopharmaceuticals given in Addendum 1 to ICRP Publication 53: 3H neutral fat and fatty acids; 14C neutral fat and fatty acids; 68Ga EDTA; and 99mTc HM-PAO, MAG3, and MIBI.

There is an integrated index to all radiopharmaceuticals treated in ICRP publications so far, including a listing of effective doses per unit activity administered to adults.

This issue of the Annals of the ICRP also includes an Addendum to ICRP Publication 72 concerning age-dependent doses to members of the public from intakes of radionuclides.
ERM proteins: From cellular architecture to cell signaling
Authors: Louvet‐Vallée Sophie
Published: July 2002

ERM (ezrin/radixin/moesin) proteins, concentrated in actin rich cell‐surface structures, cross‐link actin filaments with the plasma membrane. They are involved in the formation of microvilli, cell‐cell adhesion, maintenance of cell shape, cell motility and membrane trafficking. Recent analyses reveal that they are not only involved in cytoskeleton organization but also in signaling pathway. They play an important role in the activation of members of the Rho family by recruiting their regulators. The functions of ERM proteins are regulated by their conformational charges: the intramolecular interaction between the N‐ and C‐terminal domains of ERM proteins charges masks several binding sites, leading to a dormant protein. Different activation signals regulate ERM proteins functions by modulating these intramolecular interactions. The involvement of ERM proteins in many signaling pathways has led to study their role during development of different species.
Development of long‐lasting potentiation effects in the dentate gyrus during pentylenetetrazol kindling
Authors: Rüthrich Heinz , Grecksch Gisela , Krug Manfred
Published: July 2002

In the present study kindling was induced in rats by repeated intraperitoneal injection of pentylenetetrazol (PTZ) once every 48 h. The resulting seizure stages were registered after each PTZ application. The development of PTZ‐induced kindling and the time course of possible potentiation effects in the dentate gyrus were examined. The efficacy of perforant pathway transmission to the granule cells was tested in every second kindling session by measuring the monosynaptic evoked field potentials recorded in the dentate gyrus following single test stimuli of the perforant pathway at different times after PTZ injection in freely moving animals. The data suggest that establishment of a PTZ kindling is associated with the development of long‐lasting potentiation of the field potentials. After completion of kindling it was demonstrated that kindled rats also show a diminished learning performance. The relationship between the development of potentiation phenomena in hippocampal substructures and learning impairment is discussed.
Cathepsin B and glioma invasion
Authors: Demchik Lisa L. , Sameni Mansoureh , Nelson Kevin , Mikkelsen Tom , Sloane Bonnie F.
Published: July 2002

Increased expression of cathepsin B has been reported in a number of human and animal tumors. This has also been observed in human gliomas where increases in cathepsin B mRNA, protein, activity and secretion parallel malignant progression. In the present study, we showed that cathepsin B was directly involved in glioma cell invasion. Activity of cathepsin B was an order of magnitude higher in glioma tissue than in matched normal brain. Inhibitors of cysteine proteases reduced invasion of glioma cells in two in vitro models: invasion through Matrigel and infiltration of a glioma spheroid into a normal brain aggregate. Glioma spheroids expressed higher levels of cathepsin B than did monolayers and the ability of subclones differing in cathepsin B activity to infiltrate normal brain aggregates paralleled their cathepsin B activity. We confirmed that intracellular staining for cathepsin B occurs at the cell periphery and in cell processes and observed extracellular staining on the cell surface. In addition, we demonstrated that intracellular cathepsin B located at the cell periphery and in processes was active. The cell surface cathepsin B colocalized with areas of degradation of an extracellular matrix component. We hypothesize that the increased expression of active cathepsin B in gliomas leads to increases in invasion in vitro and in vivo and have developed a xenotransplant model in which this hypothesis can be tested.
Psychiatric Care under the Present Economic Era. An International Perspective
Authors: López-Ibor JJ
Published: July 2002

There are extrinsic as well as intrinsic limits to the resources available for health care. Therefore, economic factors have become an essential part of medical activity, leading to a disruption of the traditional doctor-patient relationships. A new economic approach of health care (ie, through managed care) and ethical attitude (to overcome the limits of managed care) are essential, especially in psychiatric practice.
GAO: Role of pharmacists in access to bupe shots and implants
Authors: Knopf Alison
Published: August 2020

Pharmacists could help with the coordination of getting buprenorphine implants or injections, which is less divertible than the oral medication, according to a report out this month from the Government Accountability Office (GAO). Providers are responsible for prescribing, storing and administering these medications, while pharmacies are responsible for dispensing them. Dispensing means giving them to the patient, who then takes them to the provider for placing the implant (a surgical procedure) or injection.
An Intrinsically Nontrimerizing Peptide Probe for Specifically Targeting Pathological Collagen in Connective Tissues
Authors: Cai Xiangdong , Wei Wenyu , Bi Yihui , Liu Zhao , Bai Zhongtian , Lei Junqiang , Xiao Jianxi
Published: August 2020

The efficient detection of pathological collagen plays an essential role in the diagnosis, prognosis, and treatment of various critical diseases such as arthritis and tumors. Peptide probes with the (Gly‐Pro‐Hyp)n sequences have been recently discovered to recognize pathological collagen; however, their staining efficacy severely suffers from their strong trimerizing tendency. An intrinsically nontrimerizing peptide probe F‐GOP‐10 has been constructed for the first time to specifically target pathological collagen. F‐GOP‐10 consists of the repetitive (Gly‐Hyp‐Pro)n sequences, and displays the single stranded conformation at 0 °C. F‐GOP‐10 has been demonstrated to selectively recognize pathological, but not intact collagen in various mouse connective tissues, and it has been successfully applied to detect pathological collagen in osteoarthritis, liver fibrosis, and various cancer tissues. Compared with the common Masson's trichrome staining method, F‐GOP‐10 displays unique selectivity to only bind to pathological collagen without the interference of intact collagen. Notably, peptide probes with different lengths of the Gly‐Hyp‐Pro triplets all maintain the single stranded conformation, demonstrating their intrinsically non‐trimerizing nature. This pure‐peptide, pure monomer probe allows the accurate concentration determination and the elimination of harmful pretreatment, which has great potential in histopathology staining and other clinical applications.
Polymer‐Based Batteries—Flexible and Thin Energy Storage Systems
Authors: Hager Martin D. , Esser Birgit , Feng Xinliang , Schuhmann Wolfgang , Theato Patrick , Schubert Ulrich S.
Published: August 2020

Batteries have become an integral part of everyday life—from small coin cells to batteries for mobile phones, as well as batteries for electric vehicles and an increasing number of stationary energy storage applications. There is a large variety of standardized battery sizes (e.g., the familiar AA‐battery or AAA‐battery). Interestingly, all these battery systems are based on a huge number of different cell chemistries depending on the application and the corresponding requirements. There is not one single battery type fulfilling all demands for all imaginable applications. One battery class that has been gaining significant interest in recent years is polymer‐based batteries. These batteries utilize organic materials as the active parts within the electrodes without utilizing metals (and their compounds) as the redox‐active materials. Such polymer‐based batteries feature a number of interesting properties, like high power densities and flexible batteries fabrication, among many more.
Exploiting a Single‐Crystal Environment to Minimize the Charge Noise on Qubits in Silicon
Authors: Kranz Ludwik , Gorman Samuel Keith , Thorgrimsson Brandur , He Yu , Keith Daniel , Keizer Joris Gerhard , Simmons Michelle Yvonne
Published: August 2020

Electron spins in silicon offer a competitive, scalable quantum‐computing platform with excellent single‐qubit properties. However, the two‐qubit gate fidelities achieved so far have fallen short of the 99% threshold required for large‐scale error‐corrected quantum computing architectures. In the past few years, there has been a growing realization that the critical obstacle in meeting this threshold in semiconductor qubits is charge noise arising from the qubit environment. In this work, a notably low level of charge noise of S0 = 0.0088 ± 0.0004 μeV2 Hz−1 is demonstrated using atom qubits in crystalline silicon, achieved by separating the qubits from surfaces and interface states. The charge noise is measured using both a single electron transistor and an exchange‐coupled qubit pair that collectively provide a consistent charge noise spectrum over four frequency decades, with the noise level S0 being an order of magnitude lower than previously reported. Low‐frequency detuning noise, set by the total measurement time, is shown to be the dominant dephasing source of two‐qubit exchange oscillations. With recent advances in fast (≈μs) single‐shot readout, it is shown that by reducing the total measurement time to ≈1 s, 99.99% two‐qubit gate fidelities can be achieved in single‐crystal atom qubits in silicon.