bluuebunny/crossref_metadata_2025 huggingface dataset:
Brain‐specific expression of transthyretin mRNA as revealed by cDNA cloning from brain
Authors: Motojima Kiyoto , Goto Sataro
Published: July 2002

cDNAs for rat transthyretin mRNA were cloned from a brain cDNA library. Sequencing analyses showed the presence of an additional 5' sequence that had not been reported for the liver mRNA corresponding to the flanking promoter region of the gene. This additional sequence was expressed only in the brain, suggesting the presence of a brain‐specific promoter.
λplac10 Transducing bacteriophage: DNA primary structure of the region of the abnormal excision
Authors: Shpakovski G.V. , Karakashly M.P. , Berlin Yu.A.
Published: July 2002

In studying molecular mechanisms of the formation of transducing bacteriophages, we have elucidated the primary structure of the phage‐bacterial DNA junction which resulted from the abnormal excision of the λplac10 phage. The process is structurally similar to the excision of the λplac5 phage and involves, in both cases, highly homological DNA stretches approximately 20 bp long, one of them being a part of the Z‐Y spacer of the lac operon and possessing a developed secondary structure. The conception of regioselective recombination as a type of illegitimate recombinational process with a certain degree of site‐specificity is suggested.
An early immunoreactive folding intermediate of the tryptophan synthase β2 subunit is a ‘molten globule’
Authors: Goldberg Michel E. , Semisotnov Gennady V. , Friguet Bertrand , Kuwajima Kunihiro , Ptitsyn Oleg B. , Sugai Shintaro
Published: July 2002

The refolding kinetics of the tryptophan synthase β2 subunit have been investigated by circular dichroism (CD) and binding of a fluorescent hydrophobic probe (ANS), using the stopped‐flow technique. The kinetics of regain of the native far UV CD signal show that, upon refolding of urea denatured β2, more than half of the protein secondary structure is formed within the dead time of the CD stopped‐flow apparatus (0.013 s). On the other hand, upon refolding of guanidine unfolded β2 the fluorescence of ANS passes through a maximum after about 1 s and then ‘slowly’ decreases. These results show the accumulation, in the 1–10 s time range, of an early transient folding intermediate which has a pronounced secondary structure and a high affinity for ANS. In this time range, the near UV CD remains very low. This transient intermediate thus appears to have all the characteristics of the ‘molten globule’ state [(1987) FEBS Lett. 224, 9‐13]. Moreover, by comparing the intrinsic time of the disappearance of this transient intermediate (t
35 s) with the time of formation of the previously characterized [(1988) Biochemistry 27, 7633‐7640] early imuno‐reactive intermediate recognized by a monoclonal antibody (t
12 s), it is shown that this native‐like epitope forms within the ‘molten globule’, before the tight packing of the protein side chains.
Proliferation of peroxisomes without simultaneous induction of the peroxisomal fatty acid βoxidation
Authors: Baumgart E. , Völkl A. , Pill J. , Fahimi H.D.
Published: July 2002

Marked proliferation of rat hepatic peroxisomes is observed after treatment with a new potent hypolipidemic drug BM 15766, as well as after bezafibrate. Whereas the relative specific activity of the peroxisomal fatty acid βoxidation system is not affected by BM 15766 it is significantly increased after bezafibrate. This is also confirmed by immunoblot analysis of individual β‐oxidation enzymes in highly purified peroxisome fractions. These observations suggest that peroxisome proliferation and the induction of the fatty acid β‐oxidation are regulated separately.
Allosteric mechanism of enhancer action?
Authors: Studitsky Vasily M.
Published: July 2002

A recurrent theme in molecular biology is ‘action at a distance’ along DNA. Why can some regulatory DNA sequences (enhancers) work at a great distance from the target of regulation? As one possible solution to this question we shall consider an allosteric mechanism of enhancer action focused on eukaryotic transcriptional enhancers.
The expression of interleukin‐6 by a rat macrophage‐derived cell line
Authors: Braciak Todd A. , Gauldie Jack , Fey George H. , Northemann Wolfgang
Published: July 2002

A stable rat macrophage‐derived cell line (RMSV1) was established by transformation of primary peritoneal exudate cells with the SV40 virus. The RMSV1 cell line was used as a model to study the regulation of the interleukin‐6 (IL6) gene expression in rate macrophages with respect to lipopolysaccharides (LPS), interleukin‐1 (IL1) and glucocorticoids. The IL6 mRNA level in RMSV1 cell lines was induced 20‐fold within 4 h by LPS, whereas IL1 had no effect. The glucocorticoids were able to inhibit completely the induction of the IL6 mRNA synthesis by LPS, indicating the negative regulation of the IL6 gene expression by glucocorticoids.
Expression of dihydropyridine‐sensitive brain calcium channels in the rat central nervous system
Authors: Chin Hemin , Smith Mark A. , Kim Hyung-Lae , Kim Hyun
Published: July 2002

We have localized dihydropyridine (DHP)‐sensitive calcium channels in rat brain by in situ hybridization and immunohistochemistry. The mRNA for the dihydropyridine‐sensitive calcium channel α1 subunit (DHPR‐B) is prominently localized in neuronal cells in the olfactory bulb, dentate gyrus, hippocampus, arcuate nucleus, paraventricular nucleus, ventromedial nucleus, cerebral cortex, superior colliculus and the cerebellar Purkinje cell layer. Strong expression of DHPR‐B mRNA was also found in the pituitary and pineal glands. DHP‐sensitive calcium channel α1 subunit distribution has also been examined immunohistochemically with polyclonal antibodies raised against synthetic peptides specific for the DHPR‐B α1 subunit protein. The results from immunohistochemistry were in good agreement with those from in situ hybridization. Thus, regional distribution and localization of DHPR‐B mRNA and α1 subunit protein in rat brain suggest that this type of DHP‐sensitive brain calcium channel may play an important role in excitation‐secretion coupling functions in the neuroendocrine system.
The solution structure of a cyclic endothelin antagonist, BQ‐123, based on1H–1H and13H–1H three bond oupling constants
Authors: Reily Michael D. , Thanabal Venkataraman , Omecinsky Diana O. , Dunbar James B. , Doherty Annette M. , DePue Patricia L.
Published: July 2002

A cyclic pentapeptide endothelin antagonist, cyclo(dTrp‐dAsp‐Pro‐dVal‐Leu), recently reported (K. Ishikawa et al., 13th Am. Pept. Symp., Cambridge MA, 1991) has been studied by NMR spectroscopy and molecular modeling. A stable structure has been determined without the use of nuclear Overhauser effects and is based primarily on homonuclear and heteronuclear three bond coupling constants. The13C‐edited TOCSY experiment is demonstrated at natural abundance and ∼30 mM peptide concentrations. Three bond13C–1H coupling constants obtained by this method are shown to reduce the ambiguity in φ angle determination which exists when only interproton coupling constants are used. Three out of four φ angles were determined uniquely by this method and the fourth was reduced to two possible values. The proline φ angle was determined to be −78° based on the3JHzHαand3JHzHβcoupling constants. Comparison of amide proton temperature dependence, chemical shifts and vicinal proton coupling constants in a 20% acetonitrile/80% water solvent mixture and in (CD3)2SO indicates that the structure is similar in both solvents.
Amino acid sequencing of a trypsin inhibitor by refined 1.6 Å X‐ray crystal structure of its complex with porcine β‐trypsin
Authors: Huang Qichen , Liu Shengping , Tang Youqi , Zeng Fuyue , Qian Ruiqing
Published: July 2002

The stoichiometric complex formed between porcine β‐trypsin and theMomordica charantia, Linn.Cucurbitaceaetrypsin inhibitor‐A (MCTI‐A) was crystallized and its X‐ray crystal structure determined using molecular replacement method. The primary sequence and topology of the inhibitor was determined by recognizing the electron density and refined to a finalRvalue of 0.167 (7.0—1.6 Å) with RMS deviation of bond lengths from standard values 0.012 Å. The sequence was compared with those obtained by other groups and was found to be similar to the squash proteinase inhibitor. Its spatial structure and the conformation of its primary binding segment from Cys‐31 (P3) to Glu‐71 (P3′) which contains the reactive scissile bond Arg‐51 C‐Ile‐61 N were also very similar with other squash family proteinase inhibitors.
Escherichia coli leucine‐responsive regulatory protein (Lrp) controls lysyl‐tRNA synthetase expression
Authors: Gazeau Myriam , Delort Florence , Dessen Philippe , Blanquet Sylvain , Plateau Pierre
Published: July 2002

Using random Tn10 insertion mutagenesis, we isolated an Escherichia coli mutant strain affected in the regulation of lysU, the gene encoding the inducible form of lysyl‐tRNA synthetase. The transposon giving rise to the altered expression of lysU was found inserted within lrp. The latter gene codes for the leucine‐responsive regulatory protein (Lrp) which mediates a global response of the bacterium to leucine. An involvement of Lrp in the regulation of lysU was searched for by using a lysU‐lacZ operon fusion. The following conclusions were reached: (i) inactivation of lrp causes an increased activity of the lysU promoter, whatever the growth conditions assayed, (ii) insertion of a wild‐type lrp gene into a multi‐copy plasmid significantly reduces lysU expression, and (iii) sensitivity of the lysU promoter to the presence of leucine in the growth medium is abolished in the lrp context.
Molecular cloning reveals the existence of a fourth γ subunit of the vertebrate brain GABAAreceptor
Authors: Harvey Robert J. , Kim Hye-Chin , Darlison Mark G.
Published: July 2002

We have isolated a cDNA, from the chicken, that encodes a fourth type of γ subunit of the vertebrate brain GABAAreceptor. The mature polypeptide (which we name γ4) displays 67%, 69% and 70% identity, respectively, to the rat γ1, γ2 and γ3 subunits. In the developing chicken brain, the γ4‐subunit mRNA is first detected at embryonic day 13; the transcript level then increases progressively during embryogenesis. In situ hybridization reveals that the γ4‐subunit mRNA is abundant in several brain regions, including the ectostriatum, nucleus rotundus and hyperstriatum ventrale, which are involved in visual processing and learning.
Endothelin‐1 increases the levels of mRNA and protein of muscarinic acetylcholine receptors and c‐fos mRNA in cerebellar granule cells
Authors: Fukamauchi Fumihiko , Chuang De-Maw
Published: July 2002

Endothelin‐1 (ET‐1) induced a time‐ and dose‐dependent increase in the levels of mRNA of m2‐ and m3‐muscarinic acetylcholine receptors (mAChRs) in cultured cerebellar granule cells. The levels of immunoprecipitable m3‐mAChR protein and total mAChR binding sites were also increased by ET‐1 treatment. The up‐regulation of m2‐ and m3‐mAChR mRNA was blocked by phorbol ester pretreatment to inhibit ET‐1‐stimulated phosphoinositide hydrolysis and was preceded by an increase in c‐fos mRNA levels. Treatments that prevented ET‐1‐induced c‐fos mRNA increase also abolished the subsequent m2‐and m3‐mAChR mRNA up‐regulation, suggesting that c‐Fos protein is involved in the ET‐1‐induced mAChR expression.
Protein kinases and protein synthesis are involved in desensitization of the plasminogen activator response of rat Sertoli cells by follicle‐stimulating hormone
Authors: Laurent-Cadoret V. , Guillou F. , Combarnous Y.
Published: July 2002

Tissue‐type plasminogen activator (tPA) secretion is a specific response of Sertoli cells to follicle‐stimulating hormone (FSH), which is lower after preincubation of the cells with low FSH concentrations because of FSH receptor/Gsprotein uncoupling. In this report, we present evidence that this desensitization induced by the lowest FSH concentrations is suppressed by specific peptidic inhibitors of endogenous PKA and PKC in permeabilized Sertoli cells. In contrast, desensitization promoted by slightly higher FSH concentrations is not mediated through PKA or PKC activation but is dependent on protein neosynthesis.
Glycotripeptides are released by yeast but not by mammalian microsomes
Authors: van Leyen Klaus , Wiedmann Martin , Wieland Felix
Published: July 2002

Glycotripeptides generated in vivo in the endoplasmic reticulum (ER) have been used as markers to assess the rate of vesicular bulk flow from the ER via the Golgi apparatus to the plasma membrane in mammalian cells. The applicability of such glycotripeptides as markers for bulk flow along this pathway has been questioned by a report on non‐vesicular release of glycotripeptides from yeast semi‐intact spheroplasts. We have therefore investigated direct release of glycotripeptides from yeast and from mammalian microsomes and report here that such release is specific to the yeast system and cannot be detected in mammalian microsomes.
Effects of transient expression of spermidine/spermine N1‐acetyltransferase in COS cells
Authors: Vargiu Cristina , Persson Lo
Published: July 2002

Mammalian spermidine/spermine N
1‐acetyltransferase (SSAT) was transiently expressed in COS cells. As compared to COS cells transfected with control vector alone, cells transfected with the expression vector containing SSAT cDNA contained lower concentrations of spermidine and spermine. The putrescine content, on the other hand, was markedly increased in the COS cells expressing large amounts of SSAT. These changes in polyamine content were most likely caused by an interconversion of spermine and spermidine into putrescine. The SSAT‐induced changes in cellular polyamine content resulted in a compensatory increase in the activities of ornithine decarboxylase and S‐adenosylmethionine decarboxylase, i.e. the enzymes catalyzing the rate‐limiting steps in polyamine biosynthesis. This is the first demonstration that a primary increase in SSAT activity will induce an interconversion‐like change in the polyamine levels and the physiological role of SSAT is most likely to protect cells against too high concentrations of spermidine and spermine.
Triton‐soluble phosphovariants of the high molecular weight neurofilament subunit from NB2a/dl cells are assembly‐competent
Authors: Shea Thomas B.
Published: July 2002

NB2a/dl cells incorporate neurofilaments (NFs) containing extensively phosphorylated high (NF‐H) molecular weight subunits into the Tritoninsoluble cytoskeleton of axonal neurites elaborated during differentiation with dibutyryl cAMP. However, immunocytochemical and biochemical analyses demonstrate the constitutive expression and extensive phosphorylation of a sizeable pool of (200 kDa) NF‐H. We examined by cell‐free analyses whether or not this Triton‐soluble NF‐H pool was assembly‐competent in cell‐free analyses. Triton‐soluble fractions from 35S‐radiolabeled NB2a/dl cells were incubated with dissociated mouse CNS Triton‐insoluble cytoskeletons that had been dissociated by treatment with 6 M urea. Following overnight dialysis to remove urea, low‐speed centrifugation to sediment Triton‐insoluble cytoskeletons resulted in the co‐sedimentation of radiolabeled NF‐H, indicating that Triton‐soluble NF‐H was capable of association with Triton‐insoluble structures. Triton‐soluble, extensively phosphorylated NF‐H from NB2a/dl cells was also capable of co‐assembling with purified NF‐L. Following high‐speed centrifugation (100,000 × g for 1 h) to sediment any oligomeric assemblies, the Triton‐soluble fraction from NB2a/dl cells was mixed with purified NF‐L that had been solubilized by 6 M urea. Following overnight dialysis to remove urea, high‐speed centrifugation sedimented both NF‐L and Triton‐soluble NF‐H from NB2a/dl cells, demonstrating that Triton‐soluble NF‐H variants are assembly‐competent. These data suggest that NF‐H variants represent precursors for NF assembly, and indicate that their assembly within NB2a/dl cells must be under temporal and spatial regulation.
HCE, a constituent of the hatching enzymes of Oryzias latipes embryos, releases unique proline‐rich polypeptides from its natural substrate, the hardened chorion
Authors: Lee Kyong-Su , Yasumasu Shigeki , Nomura Kohji , Iuchi Ichiro
Published: July 2002

HCE, a constituent protease of the hatching enzymes of Oryzias latipes embryos [1,2], releases unique proline‐rich polypeptides from its natural substrate, the hardened chorion. The polypeptides consist of repeats of Pro‐X‐Y, mainly Pro‐Glx‐X. In addition, the polypeptides contain abundant γ‐glutamyi ϵ‐lysine isopeptides which are regarded to be responsible for chorion hardening. These findings suggest that HCE recognizes specific site(s) of the chorion, releases the proline‐rich polypeptides from it, and makes the substrate accessible to LCE, another protease of the hatching enzymes.
Plasticity in the myenteric plexus of the rat ileum after long‐term sympathectomy
Authors: Milner P. , Lincoln J. , Belai A. , Burnstock G.
Published: July 2002

To investigate the effect of chronic sympathectomy on the innervation of a tissue with an extensive intrinsic component, 1‐week‐old rat pups were treated with 50 mg/kg guanethidine for 3 weeks, a treatment shown to produce complete and long‐lasting sympathectomy, and the ileum examined. Changes in the levels of noradrenaline, neuropeptide Y, calcitonin gene‐related peptide, substance P and vasoactive intestinal polypeptide in the external muscle layers containing the myenteric plexus of the ileum were determined between 6 and 20 weeks of age. After sympathectomy, noradrenaline levels were initially depleted (3% of age‐matched controls at 6 weeks,P<0.001, and 18% of age‐matched controls at 12 weeks,P<0.001), but were not significantly reduced at 20 weeks (67% of age‐matched controls). Such increases in noradrenaline content with time after sympathectomy did not occur in the mesenteric vein (levels in 20‐week‐old sympathectomized rats were 2% of the control values (P<0.001). In the myenteric plexus, catecholamine fluorescent nerve fibres were seen in the 12‐week‐old sympathectomized rats, although tyrosine hydroxylase‐immunoreactivity was absent. Guanethidine sympathectomy had no effect on the neuropeptide levels in 6‐week‐old rat ileum but there was a selective increase at 20 weeks; the levels of calcitonin gene‐related peptide and substance P were increased (×3,P<0.001 and ×1.6,P<0.05, respectively) while vasoactive intestinal polypeptide and neuropeptide Y levels were unchanged. Short‐term sympathectomy (destruction of sympathetic nerve terminals by acute 6‐hydroxydopamine treatment) had no effect on noradrenaline or peptide levels in this tissue. The changes in innervation noted after long‐term sympathectomy may represent a part of the adaptive response of the enteric nervous system which permits normal intestinal function in the absence of extrinsic neuronal inputs.
Electroconvulsive therapy: a review on indications, methods, risks and medication
Authors: Stevens A , Fischer A , Bartels M , Buchkremer G
Published: July 2002

This paper reviews and presents data of practical impact for those administering electroconvulsive therapy (ECT). In the first section, physical and physiological aspects of the stimulus as well as methods of stimulation are discussed. The second section deals with indications for ECT, efficacy and treatment modalities such as seizure duration, treatment frequency and total number of ECT applications. The last section is devoted to side effects, risks, comedication and comorbidity.
Heterologous expression of a mammalian epithelial sodium channel in yeast
Authors: Gupta Soma Sen , Canessa Cecilia M
Published: July 2002

The α and β subunits of the amiloride‐sensitive rat epithelial sodium channel (αβENaC) were expressed in the yeast Saccharomyces cerevisiae. We used a combination of yeast strains, including a mutant in the secretory pathway (sec6), and Western blotting techniques, to show that αβENaC was synthesized and targeted through the secretory system to the plasma membrane. Yeasts expressing αβENaC were more sensitive to salt than the parent strain. In addition, amiloride, a specific blocker of ENaC, was found to suppress salt sensitivity in the yeast strain expressing αβENaC.
Syk protein‐tyrosine kinase is involved in neuron‐like differentiation of embryonal carcinoma P19 cells
Authors: Tsujimura Toshiaki , Yanagi Shigeru , Inatome Ryoko , Takano Tomoko , Ishihara Itsuko , Mitsui Norihiro , Takahashi Shusuke , Yamamura Hirohei
Published: July 2002

Syk has been implicated in activated immunoreceptors to downstream signaling events in hematopoietic cells. Here we report that Syk is expressed in neuron‐like cells and involved in neuron‐like differentiation of embryonal carcinoma P19 cells. Immunoblot, RT‐PCR, and Northern analysis indicated that Syk is expressed in mouse brain, PC12 and P19 cells. In addition, Syk was found to be tyrosine phosphorylated during neuron‐like differentiation of P19 cells. Furthermore, adenovirus‐mediated overexpression of Syk induced supernumerary neurite formation and extracellular signal‐regulated kinase (ERK) activation in P19 cells. These results suggest that Syk plays an important role in signaling steps leading to ERK activation in P19 cells.
Glu‐256 is a main structural determinant for oligomerisation of human arginase I
Authors: Sabio Guadalupe , Mora Alfonso , Rangel Marı́adel Ara , Quesada Alberto , Marcos Carlos F. , Alonso Juan Carlos , Soler Germán , Centeno Francisco
Published: July 2002

One determinant that could play a role in the quaternary structure of human arginase is the pair of salt links between the strictly conserved residues R255 from one monomer and E256 from every adjacent subunit. In this work, the ionic interaction between monomers was disrupted by expressing a human arginase where Glu‐256 had been substituted by Gln. Biochemical analyses of the mutant protein showed that: (i) it shares the wild‐type kinetic parameters of the arginine substrate; (ii) E256Q arginase behaves as a monomer by gel filtration; (iii) it is drastically inactivated by dialysis in the presence of EDTA, an inhibitory effect which is reversed by addition of Mn2+; and (iv) the mutant enzyme loses thermal stability. The lack of oligomerisation for E256Q arginase and the conservation of E256 throughout evolution of the protein family suggest that this residue is involved in the quaternary structure of arginases.
Tertiary structural models for human interleukin‐6 and evaluation by a sequence‐structure compatibility method and NMR experimental information
Authors: Sumikawa Hiromi , Fukuhara Ken-ichi , Suzuki Ei-ichiro , Matsuo Yo , Nishikawa Ken
Published: July 2002

© 1997 Federation of European Biochemical Societies.
A molecular model for interfacial activation in phospholipase A2
Authors: Warwicker J.
Published: July 2002

© 1997 Federation of European Biochemical Societies.
Binding of histamine H3‐receptor antagonists to hematopoietic progenitor cells
Authors: Corbel Stéphane , Traiffort Elisabeth , Stark Holger , Schunack Walter , Dy Michel
Published: July 2002

© 1997 Federation of European Biochemical Societies.
Partial purification and characterization of a jasmonic acid conjugate cleaving amidohydrolase from the fungus Botryodiplodia theobromae
Authors: Hertel Silvia C , Knöfel Hans-Dieter , Kramell Robert , Miersch Otto
Published: July 2002

A protein preparation from the mycelium of the tropical pathogenic fungus Botryodiplodia theobromae revealed a novel peptidase activity. This enzyme was capable of cleaving conjugates of jasmonic acid with α‐amino acids. The protein was enriched 108‐fold by gel filtration, ion exchange and hydrophobic interaction chromatography. The enzyme was found to be a glycoprotein with a molecular mass of about 107 kDa. The amidohydrolase seems to be very specific with regard to (−)‐jasmonic acid and α‐amino acids with (S)‐configuration.
Improving scFv antibody expression levels in the plant cytosol1
Authors: Schouten Alexander , Roosien Jan , de Boer Jan M , Wilmink Annemiek , Rosso Marie-Noëlle , Bosch Dirk , Stiekema Willem J , Gommers Fred J , Bakker Jaap , Schots Arjen
Published: July 2002

Expression of single‐chain antibody fragments (scFvs) in the plant cytosol is often cumbersome. It was unexpectedly shown that addition at the C‐terminus of the ER retention signal KDEL resulted in significantly improved expression levels. In this report the cytosolic location of the scFv‐CK was confirmed, excluding possible mistranslocation to other subcellular compartments. It was shown that expression of several other scFvs was also improved in tobacco protoplasts. In addition expression was improved in transgenic potato. Changing from KDEL to KDEI did not affect the enhanced protein expression level. Addition of the KDEL motif is a simple and straightforward tool to stabilize in planta cytosolic expression of many scFvs.
C‐terminally deleted fragments of 40‐kDa earthworm actin modulator still show gelsolin activities
Authors: Giebing Thomas , Obermann Wolfgang M.J , Fürst Dieter , D'Haese Jochen
Published: July 2002

C‐ and N‐terminally truncated fragments of earthworm gelsolin were constructed, cloned and expressed in Escherichia coli. G‐actin‐binding properties of these fragments and their influences on the polymeric state of actin were investigated. A construct lacking a large part of the third segment [E(1–295)] supports actin nucleation similar to the complete protein and shows reduced actin fragmentation property, but is no longer Ca2+‐sensitive in its activity. The first and the second segments (E1 and E2) each contain one actin‐binding site. In contrast to human gelsolin, E1 in combination with a short N‐terminal region of E2 is not sufficient for the F‐actin‐severing activity of the protein.
Effect of mutations of murine lens αB crystallin on transfected neural cell viability and cellular translocation in response to stress
Authors: Wiesmann Kirsten E.H , Coop Audrey , Goode Derek , Hepburne-Scott Henry W , Crabbe M.James C
Published: July 2002

We examined the influence of over‐expressed native and mutant murine lens αB crystallin on the response of a murine neural cell line to heat and ionic strength shock. Native and mutant (F27R and KK174/175LL) murine αB crystallin amplicons were subcloned into a Lac‐Switch IPTG‐inducible RSV promoter eukaryotic vector, and transfected into N1E‐115 cells using lipofectin. Expression was induced maximally 8 h after addition of IPTG (optimal final concentration 1 mM) to the medium. Cells grew normally after transfection with native and mutant murine αB crystallin. We demonstrated expression of the protein using specific anti‐α crystallin antibodies. Viability under severe heat and ionic strength stress increased when cells had been transfected with native αB crystallin, but not with mutants F27R or KK174/175LL. Heat shock caused translocation of both native and mutant αB crystallins into the central region of the cells. These results show that mutations in αB crystallin that effect its chaperone‐like activity may also influence viability of N1E‐115 neural cells under stress, while not influencing the distribution of the protein within the cell.
Localization of the site for the nucleotide effectors ofEscherichia colicarbamoyl phosphate synthetase using site‐directed mutagenesis
Authors: Mora Paz , Rubio Vicente , Fresquet Vicente , Cervera Javier
Published: July 2002

Replacement by alanine of Ser‐948, Thr‐974 and Lys‐954 ofEscherichia colicarbamoyl phosphate synthetase (CPS) shows that these residues are involved in binding the allosteric inhibitor UMP and the activator IMP. The mutant CPSs are active in vivo and in vitro and exhibit normal activation by ornithine, but the modulation by both UMP and IMP is either lost or diminished. The results demonstrate that the sites for UMP and IMP overlap and that the activator ornithine binds elsewhere. Since the mutated residues were found in the crystal structure of CPS near a bound phosphate, Ser‐948, Thr‐974 and Lys‐954 bind the phosphate moiety of UMP and IMP.
Tetraspanins expressed in the embryonic chick nervous system
Authors: Perron Jeanette C. , Bixby John L.
Published: July 2002

Proteins of the tetraspanin superfamily participate in the formation of plasma membrane signaling complexes; recent evidence implicates neuronal tetraspanins in axon growth and target recognition. We used a degenerate PCR screen to identify cDNAs encoding tetraspanins expressed in the embryonic spinal cord. Two cDNAs identified apparently represent chick homologues of NAG‐2 (cnag) and CD9 (chCD9). A third clone encodes a novel tetraspanin (neurospanin). All three mRNAs are widely expressed but exhibit developmentally distinct patterns of expression in the nervous system. Both neurospanin and cnag exhibit high relative expression in nervous tissue, including brain, spinal cord and dorsal root ganglia (DRG).
Synthetic sling for genital prolapse in young women
Authors: Banu L.F.
Published: July 2002

Objective: The aim is to repair uterine prolapse in young women, utilizing synthetic tape (Merselene). It is a prospective study demonstrating an operative procedure for correction of uterine prolapse in young women. This procedure was performed in three post‐graduate teaching hospitals: Institute of Obstetrics and Gynaecology, Gandhi Hospital and Niloufer Hospital, Hyderabad, India. Method: Through an abdominal approach, the Merselene tape was fixed on the posterior surface of the uterus and anchored to the anterior longitudinal ligament, over the sacral promontory. This procedure was performed on 19 women in the age group 17–27 years, two of whom were unmarried virgins. Results: The procedure rectified the descent of the uterus and kept it anteverted. In one of the cases, exploration at cesarean section showed that the sling was intact. There were no significant intraoperative or postoperative complications. Recurrence of prolapse was not reported in any of these cases. Conclusion: This technique is a relatively simple method for correction of uterine prolapse in young women. The Merselene tape is inert, strong, flexible, effectively supports the uterus, and remains retroperitoneal.
Factors modulating supernumerary hair cell production in the postnatal rat cochleain vitro
Authors: Chardin S. , Romand R.
Published: July 2002

It has been shown in the past that extra hair cells or supernumerary cells can be produced when neonatal cochleae are maintainedin vitro. In this report, we investigated the effects of the culture methods, molecules and growth factors that are thought to be involved in cell proliferation. Quantitative studies of supernumerary hair cells were made by measuring the cell density over the entire spiral lamina at two postnatal stages: birth and 3 days after birth. With a standard feeding solution without serum, a difference in cell density was observed between the two methods of culture. Cochlear explants in a standard feeding solution supplemented with serum showed an increase of cell density only when the explantation is made at birth. Retinoic acid added to the standard feeding solution did not increase the hair cell density, while insulin induced an increase, especially at 5 μg/ml. Several growth factors were tested. Epidermal growth factor (EGF) presented a dose dependent effect with an increase of up to 30% of hair cell density that was observed in the basal region when the explantation was made at birth. Transforming growth factor‐a did not induce an increase of cell density, whereas transforming growth factor‐β presented an effect on hair cell density, with a dose dependent effect reaching 37.4% for the basal inner hair cells. Interpretation of these results is limited because of the lack of data concerning the presence of specific membrane receptors. One possibility is that insulin stimulates hair cell differentiation from existing undifferentiated cells. Another hypothesis may be related to the EGF and transforming growth factor‐β, where these molecules might induce transdifferentiation of cells by acting on the transmembrane molecules and the extracellular matrix.
Psychiatric comorbidity in alcoholics treated at an institution with both coerced and voluntary admission
Authors: Sallmén B , Nilsson LH , Berglund M
Published: July 2002

Psychiatric comorbidity in alcoholics admitted to a rehabilitation center on either a voluntary or a coerced basis were studied. A group of 104 alcoholics (37 coerced and 35 voluntarily admitted men; and 21 coerced and ten voluntarily admitted women) with a mean age (SD) of 43 ± 8 years were assessed by means of a Structural Clinical Interview in accordance with the Diagnostic and Statistical Manual (DSM)-III-R (SCID). The interview took place a mean of 7 days after admission. The frequencies of lifetime/current axis I psychiatric comorbidity (substance use disorders excluded) were 66 and 61%, respectively. Drug dependence was present in 41 and 39%, respectively, of the cases. Thirty-seven percent had a lifetime diagnosis, and 33% a current diagnosis of affective disorders, 27 and 23%, respectively, of anxiety disorders and 20 and 13%, respectively, of non-organic psychotic disorders. In a subsample of 20 subjects, depressive symptoms were found to be stable during the course of treatment. No differences in frequency of psychiatric comorbidity were found between coerced and voluntarily admitted patients (67 and 56%, respectively) or between men and women (65 and 52%, respectively). The combination of psychiatric comorbidity and drug dependence was overrepresented among the coerced patients (50 vs 16%). It was concluded that the frequencies of psychiatric comorbidity were high in the present group. The co-occurrence of alcohol dependence, drug dependence and psychiatric comorbidity was more frequent among subjects who were coercively treated.
Searching for a predictive peripherical biological model in parasuicidal behaviour
Authors: Sáiz PA , Bobes J , González MP , Cocaña I , González-Quiros P , Bousoño M
Published: July 2002

The aim of this study was to identify possible peripheral biological markers (both lipidic and hormonal) which can be easily used for the early detection of parasuicidal behaviour and to propose a predictive biological model of such behaviour. A case-control analytical study was undertaken at least 3 months after attempted suicide. Study was made of 128 patients who presented at the University General Hospital of Oviedo (Spain) with signs of self-intoxication. Lipidic and hormonal profiles were measured under basal conditions and comparison was made with a control group of healthy volunteer donors obtained from the Oviedo General Hospital blood bank. A discriminant analysis was later made with the aim of establishing a predictive biological model. This included the following variables: cholesterol, HDL-C, LDL-C and cortisol. Sensitivity and specificity were 62.5% and 65.6%, respectively. Replication and improvement of this model, through other prospective studies, could lead to the use of serum cholesterol and cortisol levels as inexpensive and readily available markers of suicide risk.
Legal protection in psychiatry: balancing the rights and needs of patients and society
Authors: Legemaate J
Published: July 2002

Attention to the legal position of psychiatric patients has greatly increased during the last 25 years. The criteria for a civil commitment have been tightened and procedural guarantees have increased. In many countries major changes and improvements in the area of mental health law took place. In this paper the main developments are discussed. Attention is given to the legal protection in case of civil commitment, to the legal position of the involuntarily admitted patient during his stay in hospital and to the relationship between patients' rights and the availability of mental health care. Furthermore the impact of legal intentions to the field of psychiatry is discussed. It is argued that legal interventions should be closely monitored, in order to prevent negative consequences. Legal interventions may be necessary, but this instrument should be used with great caution.
Ten years of transition: a review article
Authors: Lavigne Marie
Published: July 2002

Ten years after the launching of the transition process, many books and reports are offering a balance sheet of the transformation that occurred in Central and Eastern Europe and in the former Soviet Union area. The review article looks at some of them, particularly at Grzegorz W. Kolodko’s book published in 2000, “From Shock to Therapy. The Political Economy of Postsocialist Transformation”. On the basis of this book and of some recent contributions, three major issues are discussed. First, why has there been such a recession in the beginning of the transition process in all countries, and was the recession inevitable? Second, due to the dire criticisms of the standard policy applied in these countries, dubbed the “Washington consensus”, has a “post-Washington consensus” emerged? Third, as we are already engaged in the second decade of the transition process, can we state when it is bound to be over, and what role is played by the European Union enlargement in accelerating the end of transition?
EVOLUTION OF THE EVIDENCE‐BASED PAPANICOLAOU SMEAR
Authors: Mashburn Jane
Published: July 2002

Cervical cancer rates have decreased over the past century, but rates have remained relatively constant during the last decade. Evidence‐based management of the abnormal Papanicolaou (Pap) smear and the use of additional testing at the time of the Pap smear are discussed in light of the current evidence. Also explored in‐depth is the evidence base for Papanicolaou smear collection and retrieval methods, especially as it relates to their influence on the validity of the smear results. In addition, issues that need to be considered by the clinician, including the use of colposcopy, especially in light of health service implications such as the need for adequately trained personnel, increased clinical visit time, and increased cost are presented.
Generation of simulated earthquake motions considering actual earthquake phase and multiple target response spectra
Authors: Suzuki Takuya
Published: August 2020

Simulated earthquake motions whose response spectrums meet the multi‐target response spectrums are often used while designing earthquake resistant nuclear power plant facilities. Therefore, it is advantageous to utilize simulated ground motions with phase characteristics of actual earthquake ground motions in the design of nuclear power plant facilities. Several methods exist for the generation of these motions. However, these methods cannot strictly maintain phase properties of the simulated earthquake motions. A method to generate simulated earthquake motions that meets both the multi‐target response spectrum and phase property requirements does not exist. This study proposes a novel method for the generation of simulated earthquake motions considering actual earthquake phase properties and multiple target response spectra. This method is equivalent to the modal iterative error correction method, which is effective for solving inverse problems with strong discontinuities. Therefore, this study proposes an improvement of this method for spectral fitting. The outline and details of the method are described and the results of experiments performed to validate the accuracy of the method are presented.
Impact of Limited Health Literacy on Patient‐Reported Outcomes in Systemic Lupus Erythematosus
Authors: Katz Patricia , Dall’Era Maria , Trupin Laura , Rush Stephanie , Murphy Louise B. , Lanata Cristina , Criswell Lindsey A. , Yazdany Jinoos
Published: August 2020

Objective

Health disparities in patient‐reported outcomes by income and education are well documented; however, the impact of health literacy on patient‐reported outcomes has received less attention. We examined independent effects of income, education, and health literacy on patient‐reported outcomes in systemic lupus erythematosus (SLE).

Methods

Data from the California Lupus Epidemiology Study (n = 323 participants) were used. Health literacy was assessed with a validated 3‐item measure (ability to understand written information, reliance on others to understand written information, confidence in completing written forms). Patient‐reported outcomes were administered by interview in English, Spanish, Cantonese, or Mandarin. Generic and disease‐specific patient‐reported outcomes were examined using the following: 10 Patient‐Reported Outcomes Measurement Information System (PROMIS) short forms; the 8 Short Form 36 (SF‐36) health survey subscales; and 3 patient‐reported SLE disease activity and damage measures. We conducted 2 sets of multivariable analyses: the first examined education, income, or health literacy individually; the second included all 3 simultaneously. All multivariable models included age, sex, race/ethnicity, language, disease duration, and physician‐assessed disease activity and damage.

Results

More than one‐third of participants (38%) had limited health literacy (LHL), including >25% with greater than high school education. In multivariable analyses simultaneously considering education, income, and health literacy, LHL was associated with significantly worse scores on all patient‐reported outcomes except disease damage. In contrast, disparities by income were seen in only 3 PROMIS scales, 3 SF‐36 subscales, and 1 disease activity measure. No disparities by education level were noted.

Conclusion

We found significantly worse patient‐reported outcome scores among individuals with LHL, even after controlling for disease activity and damage. Whether disparities are due to actual differences in health or measurement issues requires further study.
Relationship Between Magnetic Resonance Imaging Features and Knee Pain Over Six Years in Knees Without Radiographic Osteoarthritis at Baseline
Authors: Magnusson Karin , Turkiewicz Aleksandra , Kumm Jaanika , Zhang Fan , Englund Martin
Published: August 2020

Objective

To explore whether magnetic resonance imaging (MRI) features suggestive of knee osteoarthritis (OA) are associated with presence of knee pain in possible early‐stage OA development.

Methods

We included 294 participants from the Osteoarthritis Initiative (mean ± SD age 50 ± 3 years; 50% women) with baseline Kellgren/Lawrence grade of 0 in both knees, all of whom had received knee MRIs at 4 different time points over 6 years (baseline, 24, 48, and 72 months). Using a linear mixed model (knees matched within individuals), we studied whether MRI features (meniscal body extrusion [in mm], cartilage area loss [score 0–39], cartilage full thickness loss [range 0–16], osteophytes [range 0–29], meniscal integrity [range 0–10], bone marrow lesions [BMLs] including bone marrow cysts [range 0–20], Hoffa‐ or effusion‐synovitis [absent/present], and popliteal cysts [absent/present]) were associated with knee‐specific pain as reported on the Knee Injury and Osteoarthritis Outcome Score (KOOS) questionnaire using a 0–100 scale (worst to best).

Results

The differences in KOOS knee pain score for a knee with a 1 unit higher score on MRI were the following: meniscal extrusion –1.52 (95% confidence interval [95% CI] –2.35, –0.69); cartilage area loss –0.23 (95% CI –0.48, 0.02); cartilage full thickness loss –1.04 (95% CI –1.58, –0.50); osteophytes –0.32 (95% CI –0.61, –0.03); meniscal integrity –0.28 (95% CI –0.58, 0.02); BMLs including potential cysts –0.19 (95% CI –0.55, 0.16); synovitis 0.23 (95% CI –1.14, 1.60); and popliteal cysts 0.86 (95% CI –0.56, 2.29).

Conclusion

Meniscal extrusion, full thickness cartilage loss, and osteophytes are associated with having more knee pain. Although these features may be relevant targets for future trials, the clinical relevance of our findings is unclear because no feature was associated with a clinically important difference in knee pain.
Access and Cost‐Related Nonadherence to Prescription Medications Among Lupus Patients and Controls: The Michigan Lupus Epidemiology and Surveillance Program
Authors: Minhas Deeba , Marder Wendy , Harlow Sioban , Hassett Afton L. , Zick Suzanna M. , Gordon Caroline , Barbour Kamil E. , Helmick Charles G. , Wang Lu , Lee Jiha , Padda Amrita , McCune W. Joseph , Somers Emily C.
Published: August 2020

Objective

Medication access and adherence are important determinants of health outcomes. We investigated factors associated with access and cost‐related nonadherence to prescriptions in a population‐based cohort of systemic lupus erythematosus (SLE) patients and controls.

Methods

Detailed sociodemographic and prescription data were collected by structured interview in 2014–2015 from participants in the Michigan Lupus Epidemiology and Surveillance (MILES) cohort. We compared access between cases and frequency‐matched controls and examined associated factors in separate multivariable logistic regression models.

Results

A total of 654 participants (462 SLE patients, 192 controls) completed the baseline visit; 584 (89%) were female, 285 (44%) were Black, and the mean age was 53 years. SLE patients and controls reported similar frequencies of being unable to access prescribed medications (12.1% versus 9.4%, respectively; P was not significant). SLE patients were twice as likely as controls to report cost‐related prescription nonadherence in the preceding 12 months to save money (21.7% versus 10.4%; P = 0.001) but were also more likely to ask their doctor for lower cost alternatives (23.8% versus 15.6%; P = 0.02). Disparities were found in association with income, race, and health insurance status, but the main findings persisted after adjusting for these and other variables in multivariable models.

Conclusion

SLE patients were more likely than controls from the general population to report cost‐related prescription nonadherence, including skipping doses, taking less medicine, and delaying filling prescriptions; yet, <1 in 4 patients asked providers for lower cost medications. Consideration of medication costs in patient decision‐making could provide a meaningful avenue for improving access and adherence to medications.
Patient Perspective on Using Digital Resources to Address Unmet Needs in Systemic Lupus Erythematosus
Authors: Ra Jennifer H. , Leung Jerik , Baker Elizabeth A. , Kim Alfred H. J.
Published: August 2020

Objective

The clinical variability of systemic lupus erythematosus (SLE) caused by the unpredictability of flares contributes to patients experiencing a diminished sense of social support. Digital health interventions (DHIs) have the potential to improve patients’ social support but have yet to be studied extensively in SLE. Our objective was to assess general and SLE‐specific internet usage as well as specific suggestions for SLE‐related digital resources and tools among patients with SLE at the Washington University Lupus Clinic.

Methods

Fifty‐six participants were recruited from the Washington University Lupus Clinic. Ten‐minute structured interviews consisting of multiple choice and open‐ended questions were conducted. A descriptive statistical analysis was conducted with the quantitative data, while the qualitative data were analyzed using an open coding approach.

Results

Nearly all respondents indicated having access to the internet (98.2%). Eighty percent currently use the internet for their SLE. The qualitative data indicate that patients with SLE use the internet for understanding flares and changes in their symptoms and/or laboratory results, want an increased variety of SLE information online, have a desire to exchange personal experiences and knowledge of SLE with others, and desire increased diversity in the methods of delivering digital SLE information.

Conclusion

Our findings support the continued use of DHIs for patients with SLE. We believe that these findings will aid the future development of DHIs tailored to patients with SLE.