bluuebunny/crossref_metadata_2025 huggingface dataset:
Inverse gene expression of prostacyclin and thromboxane synthases in resident and activated peritoneal macrophages1
Authors: Kuwamoto Shinsuke , Inoue Hiroyasu , Tone Yoshinori , Izumi Yoshikazu , Tanabe Tadashi
Published: July 2002

Prostacyclin and thromboxane A2 produced from prostaglandin H2 are known to be important modulators with opposite biological activities. To examine possible roles of these prostanoids in immune responses, we have studied the gene expression of prostacyclin synthase (PGIS) and thromboxane synthase (TXS) in murine resident macrophages or in macrophages elicited with casein or bacillus Calmette‐Guérin (BCG). Northern blot analyses showed that the PGIS mRNA was expressed in a decreasing order in the resident, and casein‐ and BCG‐elicited macrophages. In contrast, the TXS mRNA was expressed in an increasing order in the resident, and casein‐ and BCG‐elicited macrophages. On the other hand, the mRNA for cyclooxygenase‐2, which produces PGH2 and participates in the production of prostanoids in inflammation, was expressed in both the resident and BCG‐elicited macrophages but barely in the casein‐elicited cells. In situ hybridization analysis showed that the expression of mRNAs for PGIS and TXS was ascribable not only to the alteration of the expression levels of both mRNAs in the each macrophage but also to the changes in subpopulations of the cells expressing these mRNAs. These observations suggested that the inverse gene expression of PGIS and TXS in macrophages contributes to immune responses by modulating the relative levels of prostacyclin and thromboxane A2.
The effects of measles virus persistent infection on AP‐1 transcription factor binding in neuroblastoma cells
Authors: Fishman D , Wolfson M , Bazarski E , Segal S , Rager-Zisman B
Published: July 2002

Measles virus (MV) persistence in brain cells has broad effects on different cellular functions. We have previously shown that NS20Y clone, originally derived from C1300 neuroblastoma cells, persistently infected with MV (NS20Y/MS), displays constitutively elevated levels of c‐fos and PKC mRNAs, implying MV‐mediated effects on transcriptional regulation. Nonetheless, the mode by which virus affects the transcriptional machinery still remains obscure. In order to define this phenomenon, we studied the binding properties of major transcription factors (AP‐1 and NFκB) in NS20Y/MS cells. Using electrophoretic mobility shift approach (EMSA) with the appropriate oligonucleotide probes, we have found that the persistent MV infection does not affect NFκB binding, while the AP‐1 binding was significantly decreased. Similar inhibition was not observed in NS20Y cells acutely infected with MV. Anti‐measles antibody‐mediated restriction of viral gene expression restored AP‐1 binding, thus suggesting that measles virus proteins may affect the components of the host transcriptional machinery.
Kidney‐specific expression of a novel mouse organic cation transporter‐like protein
Authors: Mori Kiyoshi , Ogawa Yoshihiro , Ebihara Ken , Aoki Tomohiro , Tamura Naohisa , Sugawara Akira , Kuwahara Takashi , Ozaki Shoichi , Mukoyama Masashi , Tashiro Kei , Tanaka Issei , Nakao Kazuwa
Published: July 2002

Using the signal sequence trap method, we have cloned a novel 12‐membrane‐spanning transporter‐like protein, termed renal‐specific transporter (RST), from the mouse kidney. RST is a 553‐amino‐acid protein highly homologous to recently cloned organic cation transporters, e.g. it is 30% identical to rat organic cation transporter 1 at the amino acid level. Northern blot analysis has revealed that the RST gene is expressed abundantly and specifically in the kidney. In situ hybridization analysis has shown that RST gene expression is restricted to the renal proximal tubule, where various organic cations such as endogenous catecholamines and choline or clinically used cationic drugs are known to be actively excreted.
A (His)6‐tagged recombinant barley (Hordeum vulgareL.) endosperm ADP‐glucose pyrophosphorylase expressed in the baculovirus‐insect cell system is insensitive to allosteric regulation by 3‐phosphoglycerate and inorganic phosphate
Authors: Rudi Heidi , Doan Danny N.P , Olsen Odd-Arne
Published: July 2002

ADP‐glucose pyrophosphorylase from photosynthetic tissue is allosterically regulated by 3‐phosphoglycerate and inorganic phosphate. In contrast, data from our laboratory indicated that the major AGPase from barley seeds is insensitive to these regulators. Verification of this conclusion has, however, been hindered by the proteolytic degradation of the enzyme from seeds. This report characterizes the barley seed AGPase expressed in the baculovirus‐insect cell system, confirming that lack of allosteric regulation by 3‐PGA/Piis an intrinsic property of the enzyme. Purification of the enzyme was by Ni2+‐NTA agarose chromatography using a (His)6tag attached to the N‐terminus of the small AGPase subunit.
Natural ceramide is unable to escape the lysosome, in contrast to a fluorescent analogue
Authors: Chatelut Martine , Leruth Michèle , Harzer Klaus , Dagan Arie , Marchesini Sergio , Gatt Shimon , Salvayre Robert , Courtoy Pierre , Levade Thierry
Published: July 2002

Since the generation upon cell stimulation of the second messenger ceramide has been reported to occur in an endosomal/lysosomal compartment, we investigated whether ceramide formed in the lysosomes can escape this compartment. The metabolic fate of radiolabelled ceramide produced by intralysosomal hydrolysis of LDL‐associated [ceramide‐3H]sphingomyelin or [stearoyl‐1‐14C]sulfatide was examined in fibroblasts from control individuals and a patient with inborn lysosomal ceramidase deficiency (Farber disease). The behavior of this radioactive ceramide was compared to that of a fluorescent (lissamine‐rhodaminyl) ceramide analogue deriving from sulfatide degradation. While in Farber cells the natural, radiolabelled ceramide remained completely undegraded and accumulated in the lysosomes, the fluorescent derivative was rapidly converted to sphingomyelin. These findings strongly suggest that, in contrast to fluorescent derivatives, endogenous long‐chain ceramide is unable to exit from lysosomes, therefore making the lysosomal ceramide unlikely to be a biomodulatory molecule.
A novel peptide, PLAEIDGIELTY, for the targeting of α9β1‐integrins
Authors: Schneider Holm , Harbottle Richard P. , Yokosaki Yasuyuki , Kunde Jan , Sheppard Dean , Coutelle Charles
Published: July 2002

Targeting gene therapy vectors to abundant receptors on airway epithelia may allow a significant enhancement of gene delivery and thereby be of particular importance for the gene therapy of cystic fibrosis. α9β1‐Integrins are highly expressed throughout the human airway epithelia in vivo, irrespective of any particular clinical status. Aiming to improve the targeting of our non‐viral integrin‐mediated gene transfer systems to airway epithelia, we searched for a short tenascin C‐derived peptide which would bind to these integrins. By utilizing recombinant bacteriophages that display overlapping regions of the third fibronectin type III repeat of tenascin C (TNfn3), we were able to localize its α9β1‐integrin binding site to the B‐C loop of TNfn3. A synthetic Pro‐Leu‐Ala‐Glu‐Ile‐Asp‐Gly‐Ile‐Glu‐Leu‐Thr‐Tyr peptide (PLAEIDGIELTY) was shown to displace α9β1‐integrin‐expressing cells completely from binding to TNfn3. This peptide, therefore, may prove useful both for the examination of the functional importance of α9β1‐integrins in vivo and the development of gene therapy vectors or drugs targeting these integrins.
Characterization of a mammalian cDNA encoding a protein with high sequence similarity to the Drosophila regulatory protein Rhomboid
Authors: Pascall John C. , Brown Kenneth D.
Published: July 2002

The Drosophila regulatory protein Rhomboid has been demonstrated genetically to facilitate signalling within the Spitz/epidermal growth factor receptor/mitogen‐activated protein kinase pathway. Using a polymerase chain reaction (PCR)‐based strategy, we have cloned a human cDNA which encodes a protein that has high sequence similarity to Rhomboid. The encoded protein, termed rhomboid‐related protein (RRP), is predicted to contain seven transmembrane domains. Northern analysis indicates that RRP mRNA is expressed at highest levels in brain and kidney.
CH domains revisited
Authors: Stradal Theresia , Kranewitter Wolfgang , Winder Steven J , Gimona Mario
Published: July 2002

A sequence motif of about 100 amino acids, termed the `calponin homology domain' has been suggested to confer actin binding to a variety of cytoskeletal and signalling molecules. Here we analyse and compare the sequences of all calponin homology domain‐containing proteins identified to date. We propose that single calponin homology domains do not confer actin‐binding per se and that the actin‐binding motifs of cross‐linking proteins, which comprise two disparate calponin homology domains, represent a unique protein module.
Constitutive internalization and association with adaptor protein‐2 of the interleukin‐6 signal transducer gp130
Authors: Thiel Stefan , Dahmen Heike , Martens Astrid , Müller-Newen Gerhard , Schaper Fred , Heinrich Peter C. , Graeve Lutz
Published: July 2002

The transmembrane protein gp130 is the common signalling receptor subunit for the interleukin‐6 (IL‐6)‐type cytokines. It has recently been shown that the cytoplasmic domain of gp130 contains a dileucine internalization motif and that endocytosis of gp130 occurs signal‐independent. Here, we have studied whether gp130 itself undergoes constitutive internalization or whether its endocytosis is stimulated by formation of the IL‐6/IL‐6R/gp130 complex. Using two different assays, we found that gp130 is internalized independent from IL‐6/IL‐6R stimulation. In addition, we show that gp130 is constitutively associated with the cell surface adaptor complex AP‐2. Our findings strongly suggest endocytosis of gp130 to be constitutive.
Formation of titanium(IV) transferrin by reaction of human serum apotransferrin with titanium complexes
Authors: Messori Luigi , Orioli Pierluigi , Banholzer Vivien , Pais Istvan , Zatta Paolo
Published: July 2002

The reaction of human serum apotransferrin with titanium(IV) citrate under physiological conditions results in the formation of a specific bis‐titanium(IV) transferrin adduct (Ti2Tf hereafter) with two titanium(IV) ions loaded at the iron binding sites. The same specific Ti2Tf complex is formed by reacting apotransferrin with titanium(III) chloride and exposing the sample to air. The derivative thus obtained was characterized by spectroscopic techniques, including absorption, UV difference, circular dichroism and 13C NMR spectroscopies, and shown to be stable within the pH range 5.5–9.0. Surprisingly, the reaction of apoTf with titanium(IV) nitrilotriacetate (NTA) does not lead to formation of appreciable amounts of Ti2Tf, even after long incubation times, although some weak interactions of Ti(IV)NTA with apoTf are spectroscopically detected. Implications of the present results for a role of transferrin in the uptake, transport and delivery of soluble titanium(IV) compounds under physiological conditions are discussed.
Mutations of Arg198 in sarcoplasmic reticulum Ca2+‐ATPase cause inhibition of hydrolysis of the phosphoenzyme intermediate formed from inorganic phosphate
Authors: Daiho Takashi , Suzuki Hiroshi , Yamasaki Kazuo , Saino Tomoyuki , Kanazawa Tohru
Published: July 2002

Arg198 of sarcoplasmic reticulum Ca2+‐ATPase was substituted with lysine, glutamine, glutamic acid, alanine, and isoleucine by site‐directed mutagenesis. Kinetic analysis was performed with microsomal membranes isolated from COS‐1 cells which were transfected with the mutated cDNAs. The rate of dephosphorylation of the ADP‐insensitive phosphoenzyme was determined by first phosphorylating the Ca2+‐ATPase with 32Pi and then diluting the sample with non‐radioactive Pi. This rate was reduced substantially in the mutant R198Q, more strongly in the mutants R198A and R198I, and most strongly in the mutant R198E, but to a much lesser extent in R198K. The reduction in the rate of dephosphorylation was consistent with the observed decrease in the turnover rate of the Ca2+‐ATPase accompanied by the steady‐state accumulation of the ADP‐insensitive phosphoenzyme formed from ATP. These results indicate that the positive charge and high hydrophilicity of Arg198 are critical for rapid hydrolysis of the ADP‐insensitive phosphoenzyme.
Phosphorylation of the medium chain subunit of the AP‐2 adaptor complex does not influence its interaction with the tyrosine based internalisation motif of TGN38
Authors: Crump Colin M. , Banting George
Published: July 2002

Tyrosine based motifs conforming to the consensus YXXΦ (where Φ represents a bulky hydrophobic residue) have been shown to interact with the medium chain subunit of clathrin adaptor complexes. These medium chains are targets for phosphorylation by a kinase activity associated with clathrin coated vesicles. We have used the clathrin coated vesicle associated kinase activity to specifically phosphorylate a soluble recombinant fusion protein of μ2, the medium chain subunit of the plasma membrane associated adaptor protein complex AP‐2. We have tested whether this phosphorylation has any effect on the interaction of μ2 with the tyrosine based motif containing protein, TGN38, that has previously been shown to interact with μ2. Phosphorylation of μ2 was shown to have no significant effect on the in vitro interaction of μ2 with the cytosolic domain of TGN38, indicating that reversible phosphorylation of μ2 does not play a role in regulating its direct interaction with tyrosine based internalisation motifs. In addition, although a casein kinase II‐like activity has been shown to be associated with clathrin coated vesicles, we show that μ2 is not phosphorylated by casein kinase II implying that another kinase activity is present in clathrin coated vesicles. Furthermore the kinase activity associated with clathrin coated vesicles was shown to be capable of phosphorylating dynamin 1. Phosphorylation of dynamin 1 has previously been shown to regulate its interaction with other proteins involved in clathrin mediated endocytosis.
Regulation of the human apolipoprotein AIV gene expression in transgenic mice
Authors: Baralle M , Vergnes L , Muro A.F , Zakin M.M , Baralle F.E , Ochoa A
Published: July 2002

The apolipoprotein (Apo) AI‐CIII‐AIV gene cluster has a complex pattern of gene expression that is modulated by both gene‐ and cluster‐specific cis‐acting elements. In particular the regulation of Apo AIV expression has been previously studied in vivo and in vitro including several transgenic mouse lines but a complete, consistent picture of the tissue‐specific controls is still missing. We have analysed the role of the Apo AIV 3′ flanking sequences in the regulation of gene expression using both in vitro and in vivo systems including three lines of transgenic mice. The transgene consisted of a human fragment containing 7 kb of the 5′ flanking region, the Apo AIV gene itself and 6 kb of the 3′ flanking region (−7+6 Apo AIV). Accurate analysis of the Apo AIV mRNA levels using quantitative PCR and Northern blots showed that the −7+6 kb Apo AIV fragment confers liver‐specific regulation in that the human Apo AIV transgene is expressed at approximately the same level as the endogenous mouse Apo AIV gene. In contrast, the intestinal regulation of the transgene did not follow the pattern observed with the endogenous gene although it produced a much higher intestinal expression following the accepted human pattern. Therefore, this animal model provides an excellent substrate to design therapeutic protocols for those metabolic derangements that may benefit from variations in Apo AIV levels and its anti‐atherogenic effect.
Aging‐associated down‐regulation of ClC‐1 expression in skeletal muscle: phenotypic‐independent relation to the decrease of chloride conductance
Authors: Pierno Sabata , De Luca Annamaria , Beck Carol L. , George Alfred L. , Conte Camerino Diana
Published: July 2002

In order to clarify the mechanism underlying the reduction of resting membrane chloride conductance (g
Cl) during aging, the levels of mRNA encoding the principal skeletal muscle chloride channel, ClC‐1, were measured. Total RNA samples isolated from tibialis anterior muscles of aged (24–29 months old) and adult (3–4 months old) rats were examined for ClC‐1 expression using Northern blot analysis, and macroscopic g
Cl was recorded from extensor digitorum longus muscle fibers from each adult and aged rat in vitro using a two intracellular microelectrode technique. Although interindividual variability was observed, aged rats exhibited a parallel reduction of both g
Cl and ClC‐1 mRNA expression as compared to adult rats. A linear correlation exists between individual values of ClC‐1 mRNA and g
Cl. These results provide evidence that ClC‐1 is the main determinant of sarcolemmal g
Cl and demonstrate that the decrease of g
Cl observed during aging is associated with a down‐regulation of ClC‐1 expression in muscle.
Cloning of the mouse phospholipid hydroperoxide glutathione peroxidase gene1
Authors: Borchert Astrid , Schnurr Kerstin , Thiele Bernd J , Kühn Hartmut
Published: July 2002

15‐Lipoxygenases and phospholipid hydroperoxide glutathione peroxidases (PH‐GPx) are counterparts in the metabolism of hydroperoxy lipids and a balanced regulation of both enzymes appears to be important for the cellular peroxide tone regulating the expression of redox sensitive genes. In contrast to lipoxygenases the molecular biology of PH‐GPx is less well investigated. In this study we cloned the PH‐GPx cDNA from a mouse fibroblast cDNA library and the PH‐GPx gene from a mouse genomic library. The gene spans approximately 4 kb which includes 1 kb of 5′‐flanking region and consists of seven exons and six introns. The immediate promoter region does not contain a TATA box but there are binding sites for several transcription factors which also occur in the porcine gene. Our investigations provide useful tools for future targeted gene disruption studies.
Inhibition of lecithin cholesterol acyltransferase by phosphatidylcholine hydroperoxides
Authors: Davit-Spraul Anne , Thérond Patrice , Leroy Arnaud , Palmade-Rieunier Françoise , Rousset Céline , Moatti Nicole , Legrand Alain
Published: July 2002

To gain insight into the nature of the lecithin‐cholesterol acyltransferase inhibitory factor(s), we separated and collected the oxidation products from oxidized lipoproteins after lipoxygenase treatment. Isolated fractions identified by chemiluminescence, as hydroperoxides of phosphatidylcholine, were found to produce a significant reduction of lecithin‐cholesterol acyltransferase activity. The reaction kinetics of lecithin‐cholesterol acyltransferase with reconstitued high density lipoproteins were studied in the presence of 0.6 and 1.2 μM hydroperoxides of phosphatidylcholine. No significant changes in the apparent V
max were observed but a concentration‐dependent increase in slope of the reciprocal plots and in the apparent K
m values was observed with increasing hydroperoxide concentrations. These results show that the active site of lecithin‐cholesterol acyltransferase is not affected by the presence of phosphatidylcholine hydroperoxides. Nevertheless, hydroperoxides of phosphatidylcholine altered the reactivity of lecithin‐cholesterol acyltransferase for reconstitued high density lipoproteins suggesting either an alteration of the binding of lecithin‐cholesterol acyltransferase to the reconstitued high density lipoproteins or a competitive inhibition mechanism.
The hypothalamic satiety peptide CART is expressed in anorectic and non‐anorectic pancreatic islet tumors and in the normal islet of Langerhans
Authors: Jensen Per B , Kristensen Peter , Clausen Jes T , Judge Martin E , Hastrup Sven , Thim Lars , Wulff Birgitte S , Foged Christian , Jensen Jan , Holst Jens J , Madsen Ole D
Published: July 2002

The hypothalamic satiety peptide CART (cocaine and amphetamine regulated transcript) is expressed at high levels in anorectic rat glucagonomas but not in hypoglycemic insulinomas. However, a non‐anorectic metastasis derived from the glucagonoma retained high CART expression levels and produced circulating CART levels comparable to that of the anorectic tumors. Moreover, distinct glucagonoma lines derived by stable HES‐1 transfection of the insulinoma caused severe anorexia but retained low circulating levels of CART comparable to that of insulinoma bearing or control rats. Islet tumor associated anorexia and circulating CART levels are thus not correlated, and in line with this peripheral administration of CART (5–50 mg/kg) produced no effect on feeding behavior. In the rat two alternatively spliced forms of CART mRNA exist and quantitative PCR revealed expression of both forms in the hypothalamus, in the different islet tumors, and in the islets of Langerhans. Immunocytochemistry as well as in situ hybridization localized CART expression to the somatostatin producing islet D cell. A potential endocrine/paracrine role of islet CART remains to be clarified.
The interaction between the archaeal elongation factor 1α and its nucleotide exchange factor 1β
Authors: Raimo Gennaro , Masullo Mariorosario , Bocchini Vincenzo
Published: July 2002

In Sulfolobus solfataricus the binding of the exchange factor 1β (SsEF‐1β) to SsEF‐1α·GDP displaces the nucleotide and the SsEF‐1α·SsEF‐1β complex is formed. The complex itself is stable, but it dissociates upon the addition of GDP or Gpp(NH)p but not ATP. Since the rate of the formation of the SsEF‐1α·SsEF‐1β complex is significatively slower than the rate of the nucleotide exchange catalyzed by SsEF‐1β it can be inferred that in vivo the GDP/GTP exchange reaction proceeds via an SsEF‐1α–SsEF‐1β interaction without involving the formation of a stable binary complex as an intermediate.
Effect of mutagenesis at serine 653 of Arabidopsis thaliana acetohydroxyacid synthase on the sensitivity to imidazolinone and sulfonylurea herbicides
Authors: Lee Yu-Ting , Chang Alan K , Duggleby Ronald G
Published: July 2002

Resistance to sulfonylurea and imidazolinone herbicides can occur by mutations in acetohydroxyacid synthase (EC 4.1.3.18). Changing serine 653 to asparagine is known to cause insensitivity to imidazolinones but not to sulfonylureas. Here, S‐653 of the Arabidopsis thaliana enzyme was mutated to alanine, threonine and phenylalanine. The purified mutated enzymes resemble wild‐type in their enzymatic properties. The threonine and phenylalanine mutants are imidazolinone‐resistant and the latter is also slightly sulfonylurea‐resistant. The alanine mutant remains sensitive to both herbicides. The results suggest that the β‐hydroxyl group is not required for imidazolinone binding and that the size of the side‐chain determines resistance.
Identification of three novel members of the calcium‐dependent chloride channel (CaCC) family predominantly expressed in the digestive tract and trachea1
Authors: Agnel Magali , Vermat Thierry , Culouscou Jean-Michel
Published: July 2002

Three novel human sequences showing striking homology to the recently described bovine Ca2+/calmodulin kinase II‐dependent epithelial chloride channel bCaCC have been identified in an expressed sequence tags database. Full‐length clones were isolated using a 5′ RACE approach. The encoded predicted proteins display 65% overall homology to bCaCC. Tissue expression patterns of the corresponding genes, designated as hCaCC‐1, ‐2 and ‐3, appear to be highly restricted, with the first two genes primarily expressed in the digestive tract. Another original feature as compared to the CaCC family members is the fact that hCaCC‐2 also shows expression in the brain. Taken together these findings demonstrate the existence of several CaCC‐like genes in humans, some of which display distinct tissue specificity patterns within the CaCC subfamily of chloride channels.
A novel human UDP‐N‐acetyl‐D‐galactosamine:polypeptide N‐acetylgalactosaminyltransferase, GalNAc‐T7, with specificity for partial GalNAc‐glycosylated acceptor substrates
Authors: Bennett Eric Paul , Hassan Helle , Hollingsworth Michael A. , Clausen Henrik
Published: July 2002

A novel member of the human UDP‐N‐acetyl‐D‐galactosamine:polypeptide N‐acetylgalactosaminyltransferase gene family, designated GalNAc‐T7, was cloned and expressed. GalNAc‐T7 exhibited different properties compared to other characterized members of this gene family, in showing apparent exclusive specificity for partially GalNAc‐glycosylated acceptor substrates. GalNAc‐T7 showed no activity with a large panel of non‐glycosylated peptides, but was selectively activated by partial GalNAc glycosylation of peptide substrates derived from the tandem repeats of human MUC2 and rat submaxillary gland mucin. The function of GalNAc‐T7 is suggested to be as a follow‐up enzyme in the initiation step of O‐glycosylation.
S. pombe Pbh1p: an inhibitor of apoptosis domain containing protein is essential for chromosome segregation
Authors: Rajagopalan Srividya , Balasubramanian Mohan K
Published: July 2002

Proteins containing the baculovirus inhibitor of apoptosis repeats (BIR domains) have been identified in a wide range of species. BIR domain containing proteins are thought to inhibit caspases and thereby cause inhibition of apoptosis. A BIR domain containing protein has been recently identified by the Schizosaccharomyces pombe genome sequencing project. However, caspase‐like proteins have not been found in yeasts, suggesting that the BIR domain containing proteins might play a fundamental role in cell regulation, in addition to their well‐characterized role in inhibition of apoptosis. In this study, we have characterized Pbh1p, an S. pombe BIR domain containing protein. Construction and analysis of a null mutant in pbh1
+ revealed that pbh1
+ is essential for cell viability. Moreover, cells devoid of Pbh1p are defective in chromosome condensation and chromosome segregation. Thus, proper chromosome segregation requires the function of Pbh1p. Over‐production of Pbh1p led to abnormalities in mitosis and cytokinesis, suggesting that the levels of Pbh1p are important for regulation of mitosis and cytokinesis.
Second and early third trimester pregnancy termination by extra‐amniotic balloon and intracervical PGE2
Authors: Liu H.‐S. , Yu M.‐H. , Chang Y.‐K. , Chu T.‐Y.
Published: July 2002

Objective: The aim of the study was to investigate the efficacy and safety of an alternative modality using extra‐amniotic balloon and intracervical prostaglandin (PG) E2 for termination of second and early third trimester pregnancies. Method: Thirty‐three pregnant women scheduled for legal termination of pregnancy at 15–32 weeks' gestation were included in the study. Each case received extra‐amniotic balloon containing 500–800 ml normal saline and two PGE2 tablets inserted into the cervical canal. Results: All 33 patients achieved a successful termination. The mean induction‐to‐abortion interval was 12.85 h. There was no significant difference in induction‐to‐abortion interval between second trimester group and third trimester group. However, the mean duration of balloon distention was longer in the second trimester group. There were no severe complications. Conclusion: The combined use of extra‐amniotic balloon and intracervical PGE2 tablets is effective, safe and convenient for termination of second and early third trimester pregnancy.
Research opportunities in service process design
Authors: Hill Arthur V , Collier David A , Froehle Craig M , Goodale John C , Metters Richard D , Verma Rohit
Published: July 2002

This paper presents an overview of the new issues and research opportunities related to four service operations design topics—the design of retail and e‐tail service processes, design of service processes involving waiting lines and workforce staffing, service design for manufacturing, and re‐engineering service processes. All four topics are motivated by new technologies (particularly web‐based technologies) and require a multi‐disciplinary approach to research. For each topic, the paper presents an overview of the topic, the relevant frameworks, and a discussion of the research opportunities.
Expression of a newly defined tumor‐rejection antigen SART3 in musculoskeletal tumors and induction of HLA class I‐restricted cytotoxic T lymphocytes by SART3‐derived peptides
Authors: Tsuda Naotake , Murayama Kumiko , Ishida Hyota , Matsunaga Kazuko , Komiya Setsuro , Itoh Kyogo , Yamada Akira
Published: July 2002

We recently reported that a SART3 tumor‐rejection antigen possessing tumor epitopes is capable of inducing HLA class I‐restricted and tumor‐specific cytotoxic T lymphocytes in cancer patients. We studied the expression of the SART3 protein in musculoskeletal tumors to find a molecule for potential use in tumor‐specific immunotherapy. The SART3 was detected at protein levels in 100% of the osteosarcoma cell lines (n = 20), in 50% of the musculoskeletal tumor tissue specimens (n = 32), and at notable levels in 67% of osteosarcoma tissues (n = 9) and malignant fibrous histiocytosis tissues (n = 9), respectively. SART3‐derived peptides at positions 109‐118 and 315‐323 induced HLA‐A24‐restricted tumor‐specific cytoxic T lymphocytes from peripheral blood mononuclear cells of patients with osteosarcoma or malignant fibrous histiocytosis. These peptide‐induced cytotoxic T lymphocytes recognized HLA‐A24+ SART3+ osteosarcoma cells but not HLA‐A24− or SART3− cells. These results suggest that the SART3 protein and its derived peptides could be molecules appropriate for use in specific immunotherapies for approximately 60% of HLA‐A24+ patients with osteosarcoma or malignant fibrous histiocytosis. © 2001 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.
Expression of 5‐HT2A, 5‐HT2B and 5‐HT2C receptors in the mouse embryo
Authors: Lauder Jean M. , Wilkie Mary Beth , Wu Chun , Singh Sujay
Published: July 2002

Expression patterns of 5‐HT2A, 5‐HT2B and 5‐HT2C receptors during mouse embryogenesis were investigated using highly specific monoclonal antibodies. Differential and overlapping spatio‐temporal patterns of 5‐HT2A, 5‐HT2B and 5‐HT2C receptor immunoreactivity were observed during active phases of morphogenesis of a variety of embryonic tissues, including neuroepithelia of brain and spinal cord, notochord, somites, cranial neural crest, craniofacial mesenchyme and epithelia, heart myocardium and endocardial cushions, tooth germs, whisker follicles, cartilage and striated muscle. The functional significance of these receptors was tested by exposing headfold stage mouse embryos to different subtype‐selective 5‐HT2 receptor antagonists for 2 days in whole embryo culture. The most potent was the pan 5‐HT2 receptor antagonist ritanserin, which has high affinity for the 5‐HT2B receptor. Ritanserin caused 100% malformed embryos at a dose of 1 μM. The 5‐HT2A/2C receptor antagonist mianserin also caused a significant number of malformed embryos, but only when used at a 10 fold higher dose (10 μM). Ketanserin, which primarily targets 5‐HT2A receptors, did not cause a significant number of malformed embryos at any dose tested. Together with previous evidence that 5‐HT acts as an important morphoregulatory signal during mouse embryogenesis, present evidence for the early and continued expression of functional 5‐HT2 receptors throughout gestation raises the possibility that psychotropic drugs taken during pregnancy could interfere with developmental actions of 5‐HT during prenatal development of neural and non‐neural tissues.
Developmental and aging aspects of the cholinergicinnervation of the olfactory bulb
Authors: Durand Marielle , Coronas Valérie , Jourdan François , Quirion Rémi
Published: July 2002

The olfactory bulb is a limbic paleocortex which receives monosynaptic sensoryafferents from the olfactory mucosa, and a strong direct cholinergic input from the basal forebrain.This review focuses on the rat olfactory bulb as a suitable model to study cholinergicinvolvements in cortical processing, during development, adulthood and aging. Anatomical andbiochemical data show that cholinergic influences upon the bulbar neuronal network are exertedthrough several types of target cells and receptors (muscarinic and nicotinic). Functional dataindicate that cholinergic afferents to the olfactory bulb are involved in local events related toolfactory learning. Neurodegenerative disorders such as Alzheimers disease involve earlyolfactory deficits and typical histopathological lesions in the olfactory bulb. In summary, with itsexclusively extrinsic cholinergic innervation and direct sensory input, the rat olfactory bulb offersthe opportunity to study the cellular and molecular mechanisms of cholinergic influences oncortical processing, in both normal and pathological conditions.
Personality traits and smoking in patients with obsessive-compulsive disorder
Authors: Bejerot S , von Knorring L , Ekselius L
Published: July 2002

As opposed to other psychiatric populations, subjects with obsessive-compulsive disorder (OCD) smoke less than the general population. The present study aims at further investigating the relationship between smoking in OCD subjects and personality traits.

Sixty-four subjects with OCD were interviewed concerning their smoking habits. Personality traits were evaluated using the Karolinska Scales of Personality, and specific obsessive-compulsive personality traits were elicited through self-report questionnaires.

Non-smokers were more easily fatigued, more inclined to worry, more remorseful, less self-confident, less impulsive and became uneasy more frequently when urged to speed up, than smokers with OCD.

Additionally, non-smokers fulfilled significantly more obsessive-compulsive personality disorder criteria as compared to the smokers (P< 0.001).

We propose a clinical subtype of OCD related to non-smoking, psychasthenia, anxiety, and pronounced obsessive-compulsive personality disorder traits.
Associations of past conduct disorder with personality disorders in ‘non-psychotic’ psychiatric inpatients
Authors: Dowson J.H. , Sussams P. , Grounds A.T. , Taylor J.C.
Published: July 2002

The aim was to investigate associations of a history of features of DSM-III-R conduct disorder (CD) with features of DSM-III-R personality disorders (PDs) and psychopathy, in inpatient psychiatric practice. Fifty-six psychiatric inpatients, without a history of specified ‘psychoses’, were assessed by the SCID structured interview for DSM-III-R PDs and the ‘Psychopathy Checklist Revised’ (PCL-R).

In a sample in which 59% had borderline PD, significant associations between a history of CD criteria and the adult features of antisocial PD (APD) were relatively specific compared with other PDs, but were weaker in women. However, significant correlations between the number of positive CD criteria and PCL-R scores were similar in both genders. The relatively specific associations between CD and adult features of APD are likely to be relevant to psychiatric patients who show various presentations of PD, if these include some adult features of APD. The findings inform the understanding of the development and classification of PDs.
Catalyst Free Silicone Sealants That Cure Underwater
Authors: Bui Robert , Brook Michael A.
Published: April 2020

Silicone sealants and adhesives are widely used to prevent the ingress of water. However, silicones must normally be cured in air, as excess water inhibits or prevents cure from occurring. It is reported that aqueous solutions of the aliphatic aldehydes glutaraldehyde, glyoxal or, particularly, formaldehyde rapidly react without catalysts with a variety of aminopropyl‐modified silicone polymers to give silicone elastomers, even underwater. These products, whose properties are readily tailored by controlling the density of amino groups in the starting materials, may be 3D printed, or used both as adhesives and sealants in air or water.
Significance of Selective Protein Degradation in the Development of Novel Targeted Drugs and Its Implications in Cancer Therapy
Authors: Yang Jie , Wang Qiaoli , Feng Guo‐Kai , Zeng Mu‐Sheng
Published: April 2020

The evolution of traditional chemotherapeutic drugs to targeted drugs has led to major changes in cancer treatment. However, it remains difficult to develop effective targeted drugs that can act against “undruggable” proteins, including some well‐known oncoproteins such as KRas. Moreover, mutations of target genes limit the use of targeted drugs. Although some strategies such as RNA interference and DNA editing have been adopted to solve these problems, their clinical use remains challenging. Therefore, new strategies are urgently needed to meet the dilemmas encountered in the use of targeted drugs. As protein degradation is a rapid and well‐regulated biological process in cells, targeting protein degradation by inducing cellular protein degradation machinery, regardless of the status of the target, is an attractive idea for developing novel targeted drugs. This review summarizes recent advances in targeted protein degradation, with a focus on the current reagents and strategies used for inducing selective degradation of target proteins to provide clues for the development of novel anticancer drugs and cancer therapies.
The immense and continuing challenge of urban governance: Developments in Australia and across Greater China
Authors: Podger Andrew , Woods Michael , Su Tsai‐tsu
Published: April 2020

The article describes the long history of urbanisation, including developments in four jurisdictions – three in Greater China (the People's Republic of China, Hong Kong and Taiwan) and Australia. While each has its own history, all now face challenges associated with continuing urbanisation and the growth of very large cities. There are broadly similar economic forces at work, but differences in scale and differences in the “growth machines” involved and the institutional arrangements within which urban governance operates. The article explains the growing importance internationally of urban governance and the growing interest in the “performance” of cities. The authors then set out two case studies describing developments in urban governance around Shanghai and around Sydney. In both cases, the scale of big city growth is demanding both horizontal and vertical coordination, and processes that facilitate citizens and civil society participation. While the very different institutional arrangements in these jurisdictions have led to different urban governance arrangements, there are common challenges that have led to some common governance developments such as having a lead government with citywide and comprehensive public service responsibilities, and also having the national government play an increasingly active role in support.
Catalyst design using artificial intelligence: SO2 to SO3 case study
Authors: Boucheikhchoukh Ariel , Thibault Jules , Fauteux‐Lefebvre Clémence
Published: April 2020

Catalyst design is key to the improvement of chemical process efficiency. The required work for the development of new catalysts can be supported through the proper application of artificial intelligence to identify optimal compositions. A generic methodology for the application of machine learning to catalysis research is therefore outlined in this work. The catalytic oxidation of SO2 was used to exemplify the first iteration of this methodology. 1784 data points from 31 published papers were compiled into a databank. The inlet SO2 concentration ranged from 0 to 66 mol%. An artificial neural network (ANN) was trained on the databank in order to predict SO2 conversion based on the catalyst composition and the reactor operating conditions (temperature, pressure, catalyst mass: volumetric flowrate ratio (w/v), and feed composition). The model achieved a root‐mean‐square error of 6.6%. A preliminary screening step identified 3:1 V‐Mg/SiO2 catalysts as exhibiting high conversion at 648 K. A multi‐objective optimization was then performed on a single catalyst to identify solutions exhibiting high conversion and high productivity at 648 K while minimizing the catalyst cost. The optimal solution was predicted to be a 2.9 wt% V/0.2 wt% Mg/SiO2 catalyst operating at a w/v of 7.49 kg‐cat · s/m3 STP, achieving 100% SO2 conversion with a material cost among the bottom third of cost values. Artificial intelligence can then be employed to extract useful knowledge from published catalytic data and orient future search for novel catalyst development.
Electrophysiological Studies of GABAA Receptors Using QPatch II, the Next Generation of Automated Patch‐Clamp Instruments
Authors: Schupp Melanie , Park Sung Hoon , Qian Binbin , Yu Weifeng
Published: April 2020

Ligand‐gated ion channels (LGICs) are a group of diverse ion channels that are gated by ligands and play important roles in normal physiological and pathological conditions. Many of them are drug targets that have been pursued, are being pursued, and will likely be pursued in the future by pharmaceutical companies and academic groups for a variety of diseases. One of those LGICs is the GABAA receptor, a heterooligomeric chloride channel that can be blocked and modulated at various sites. In order to study the receptor's functional response to compounds, the manual patch‐clamp method provides a detailed but low‐throughput electrophysiological characterization. QPatch II, a next‐generation automated patch clamp machine that was recently developed by Sophion Bioscience, provides an automated electrophysiological study of ion channels. In this article, we use the GABAA receptor as an example for studying LGICs and describe two detailed protocols for using QPatch II to carry out pharmacological studies on the receptor. © 2020 by John Wiley & Sons, Inc.

Basic Protocol 1: Ligand concentration‐response experiment (GABAA receptor) on QPatch II

Alternate Protocol: Non‐cumulative ligand concentration‐response experiment (GABAA receptor) on QPatch II

Support Protocol 1: Cell culture of HEK293‐hGABAA (α5β3γ2)

Support Protocol 2: Data analysis for Basic Protocol 1

Support Protocol 3: Data analysis for Alternate Protocol

Basic Protocol 2: Antagonist dose‐response experiment (GABAA receptor) on QPatch II

Support Protocol 3: Data analysis for Basic Protocol 2
A case report of papillary thyroid carcinoma dedifferentiated to squamous cell carcinoma presenting as a lung metastasis: A potential diagnostic pitfall
Authors: Gadde Ramya , Tafe Laura J. , Tsapakos Michael J. , Liu Xiaoying
Published: April 2020

Papillary thyroid carcinoma (PTC) tall cell variant (TCV) with squamous dedifferentiation is a rare entity. We present a case of 90‐year‐old woman who initially had a 2.8 cm conventional PTC in right lobe of thyroid who, couple decades later, had metastatic dedifferentiated PTC to right neck lymph nodes level II and IV with tall cell features; to right level IV and V lymph nodes with tall cell and squamous components, which recently presented exclusively as squamous cell carcinoma (SCC) metastasizing to lung. The squamous component in the lymph node and SCC in the lung were both positive for squamous marker p63 and PTC markers TTF1, PAX‐8 and BRAF V600E while negative for thyroglobulin and p16. The papillary component was positive for TTF‐1, BRAF V600E and P63 (majority); negative for thyroglobulin and p16. Final diagnoses were rendered based on combination of cytological features and immunohistochemical profiles. This report highlights the utilization of current biomarkers to distinguish between metastatic dedifferentiated PTC with squamous features and primary lung SCC, as well as the importance of recognizing this rare entity.
Client Experiences of Mindfulness Meditation in Counseling: A Qualitative Study
Authors: O'Brien Veronica , Likis‐Werle Elizabeth
Published: April 2020

Using qualitative methodology, the authors explored the experiences of 8 clients who received a 5‐minute mindfulness meditation (MM) as implemented by 6 counseling students during a counseling session. Themes that emerged included (a) variations in individual experiences; (b) mental, physical, and emotional components of the experience; (c) perceptions of MM; (d) preferences for MM; and (e) implications for continued practice of MM.
The SAR Matrix Method and an Artificially Intelligent Variant for the Identification and Structural Organization of Analog Series, SAR Analysis, and Compound Design
Authors: Yoshimori Atsushi , Bajorath Jürgen
Published: April 2020

The SAR Matrix (SARM) approach was originally conceived for the systematic identification of analog series, their structural organization, and graphical structure‐activity relationship (SAR) analysis. For structurally related series, SARMs also produce virtual candidate compounds. Hence, SARM represents a unique computational approach establishing a direct link between SAR visualization and compound design. The SARM data structure is reminiscent of R‐group tables and hence easily accessible from a chemical perspective, although the underlying algorithmic basis is complex. The SARM concept has been extended in different ways to further increase its analytical and design capacity. While the efforts were largely driven from a research perspective, they have also increased the utility for practical applications. Among others, extensions include approaches for SARM‐based compound activity prediction, the generation of a large SARM database for analog searching, and the design of a deep learning architecture for advanced analog design taking chemical space information for target families into account. Herein, the SARM approach and its extensions are discussed within their scientific context.
High Prevalence of Obesity in Severe Acute Respiratory Syndrome Coronavirus‐2 (SARS‐CoV‐2) Requiring Invasive Mechanical Ventilation
Authors: Simonnet Arthur , Chetboun Mikael , Poissy Julien , Raverdy Violeta , Noulette Jerome , Duhamel Alain , Labreuche Julien , Mathieu Daniel , Pattou Francois , Jourdain Merce ,   the LICORN and the Lille COVID‐19 and Obesity study group
Published: April 2020

Objective

The COVID‐19 pandemic is rapidly spreading worldwide, notably in Europe and North America where obesity is highly prevalent. The relation between obesity and severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2) has not been fully documented.

Methods

This retrospective cohort study analyzed the relationship between clinical characteristics, including BMI, and the requirement for invasive mechanical ventilation (IMV) in 124 consecutive patients admitted in intensive care for SARS‐CoV‐2 in a single French center.

Results

Obesity (BMI > 30) and severe obesity (BMI > 35) were present in 47.6% and 28.2% of cases, respectively. Overall, 85 patients (68.6%) required IMV. The proportion of patients who required IMV increased with BMI categories (P < 0.01, χ2 test for trend), and it was greatest in patients with BMI > 35 (85.7%). In multivariate logistic regression, the need for IMV was significantly associated with male sex (P < 0.05) and BMI (P < 0.05), independent of age, diabetes, and hypertension. The odds ratio for IMV in patients with BMI > 35 versus patients with BMI < 25 was 7.36 (1.63‐33.14; P = 0.02).

Conclusions

The present study showed a high frequency of obesity among patients admitted in intensive care for SARS‐CoV‐2. Disease severity increased with BMI. Obesity is a risk factor for SARS‐CoV‐2 severity, requiring increased attention to preventive measures in susceptible individuals.