bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of Clinical Stage T2 Prostate Cancer: Do Current Subclassifications Represent an Improvement?
Authors: Cagiannos Ilias , Graefen Markus , Karakiewicz Pierre I. , Ohori Makato , Eastham James A. , Rabbani Farhang , Fair William , Wheeler Thomas M. , Hammerer Peter G. , Haese Alexander , Erbersdobler Andreas , Huland Hartwig , Scardino Peter T. , Kattan Michael W.
Published: July 2002

PURPOSE: The purpose of this study was to determine whether the extent of palpable cancer within the prostate predicts outcome after radical prostatectomy.

PATIENTS AND METHODS: We combined prospectively collected data on 1,755 consecutive clinical stage T2 patients treated with radical prostatectomy alone at four institutions. According to the 1992 American Joint Committee on Cancer tumor-node-metastasis system, 645 (37%) were T2a, 758 (43%) were T2b, and 352 (20%) were T2c. Kaplan-Meier and proportional hazards regression analyses were performed on the 1992 and 1997 T2 subclassifications. After controlling for the effects of prostate-specific antigen (PSA) and biopsy Gleason sum, the two staging systems were compared for their ability to predict recurrence-free survival (RFS). Adjusted RFS curves were constructed using the corrected group prognosis method.

RESULTS: Follow-up ranged from 1 to 166 months (median, 26 months). Cancer recurred in 417 (24%) of the T2 patients. The 1992 (P = .005) but not the 1997 (P = .100) T2 subclassification predicted outcome after controlling for PSA and Gleason sum. After covariate adjustment, RFS was 7% higher at 5 years in the 1992 T2a subcategory relative to the T2b subcategory.

CONCLUSION: The 1992 American Joint Committee on Cancer system is superior to the 1997 system, and the former adds prognostic information to a model containing pretreatment PSA and Gleason sum. These results suggest that 1992 T2 subclassification derived from palpable findings improves prognostication over the 1997 subclassification.
Studies on the half life time of rat liver transfer RNA species
Authors: Hanoune Jacques , Agarwal M.K.
Published: July 2002

The tRNA fraction, extracted from very high speed supernatant fluid, from livers of rats injected with 3H‐orotic acid, attained maximum specific activity after a little over 24 hr and, thereafter, decayed with an apparent half life of 5 days. This behaviour of tRNA was indistinguishable from that of liver rRNA and the acid soluble pool. Chromatography of tRNA, doubly labelled during a period of short synthesis and of prolonged decay, on a BD cellulose column, indicated that individual tRNA species turn over at a constant rate with respect to one another.
Cooperative non‐specific DNA binding of the N‐terminal core of the cyclic AMP receptor protein of Escherichia coli and its modulation by cyclic AMP
Authors: Clore G.Marius , Gronenborn Angela M. , Davies R.Wayne
Published: July 2002

The non‐specific DNA binding of CRP and its N‐terminal core, αCRP, to a 298 base pair DNA fragment, in the presence and absence of cAMP, has been studied using the nitrocellulose filter binding technique and analysed quantitatively using the theory of Clore et al. [J. Mol. Biol. (1982) 155, 447–466]. It is shown that both CRP and αCRP bind cooperatively to DNA. At an ionic strength of 100 mM and pH 7.5, the intrinsic equilibrium association constant for the binding of αCRP to DNA is ∼ 10‐times smaller than that for CRP, but the cooperativity parameter is ∼ 17‐times larger for αCRP than CRP. cAMP exerts its effect soley on the intrinsic equilibrium constant and does not alter the cooperativity. In the case of αCRP, cAMP reduces the intrinsic equilibrium association constant by a factor of 3, in contrast to the case of CRP where cAMP increases it by a factor of 3. The possible location of the DNA binding site present in the N‐terminal core of CRP is discussed in the light of crystallographic data on the cAMP·CRP complex [McKay (1982) J. Biol. Chem. 257, 9518–9524].
Killer toxin from Hansenula mrakii selectively inhibits cell wall synthesis in a sensitive yeast
Authors: Yamamoto Teturo , Hiratani Tamio , Hirata Hajime , Imai Mitsunobu , Yamaguchi Hideyo
Published: July 2002

Hansenula mrakii secretes extracellularly a killer toxin which kills sensitive Saccharomyces cerevisiae. In protoplasts of this yeast, the killer toxin selectively inhibited the synthesis of alkali‐insoluble acid‐insoluble polysaccharides consisting mainly of β‐glucan, but did not inhibit either the synthesis of other cell wall polysaccharides, such as mannan, chitin and alkali‐insoluble acid‐soluble polysaccharides, or the synthesis of protein. Consistent with these results, the toxin was inhibitory to the β‐(1,3)‐glucan synthetase activity of a cell‐free extract from sensitive S. cerevisiae.
Nerve growth factor induces mast cell degranulation without changing intracellular calcium levels
Authors: Mazurek N. , Weskamp G. , Erne P. , Otten U.
Published: July 2002

Nerve growth factor (NGF) induces degranulation of rat peritoneal mast cells (RPMC) in a dose‐dependent manner, providing direct evidence for its action on non‐neuronal tissues. Activation of RPMC by NGF depends on lysophosphatidylserine and extracellular calcium. NGF‐mediated RPMC degranulation is not coupled to a transient increase in intracellular free calcium ([Ca2+]). It is suggested that NGF has a unique mode of action independent of ([Ca2+])i and presumably also without involving protein kinase C activation as indicated by the effects of phorbol esters and NGF on antigen‐evoked [Ca2+]i rise.
Pulsed magnetic fields alter the cell surface
Authors: Goodman E.M. , Sharpe Paul T. , Greenebaum B. , Marron Michael T.
Published: July 2002

Pulsed magnetic fields (PMFS) are routinely used in the medical community to facilitate bone repair in clinical cases of non‐union or pseudarthoses [(1984) Orth. Clin. No. Am. 15, 61‐87]. Although this therapeutic regimen appears to be reasonably effective, the mechanism of action between specific PMFs and the target tissue remains unknown. Adding urgency to the need to understand the mechanism are a wide number of reports that have appeared which demonstrate that PMFs similar to those in clinical use can alter many basic physiological functions. We report that a 24 h exposure to PMFs alters the cell surface of Physarum polycephalum amoebae. Further, using the technique of aqueous two‐phase partitioning, we present evidence for individual magnetic and electric field, cell surface effects.
A new type of functional VIP receptor has an affinity for helodermin in human SUP‐T1 lymphoblasts
Authors: Robberecht Patrick , Waelbroeck Magali , De Neef Philippe , Tastenoy Michèle , Gourlet Philippe , Cogniaux Jacqueline , Christophe Jean
Published: July 2002

A new type of VIP receptor was characterized in human SUP‐T1 lymphoblasts. The order of potency of unlabeled peptides in the presence of [125I]helodermin, was: helodermin(1–35)‐NH2 = helodermin(1–27)‐NH2 > helospectin > VIP = PHI > [D‐Ser2]VIP > [D‐Asp3]VIP > [D‐His1]VIP ⩾ [D‐Ala4]VIP 2̆ secretin = GRF. This specificity was distinct from that of all VIP receptors described so far in that: (i) the affinity for helodermin (Kd = 3 nM) was higher than that of VIP (Kd = 15 nM) and PHI (Kd = 20 nM); and (ii) position 4 played an important role in ligand binding. The labeled sites were likely to be functional receptors as adenylate cyclase in crude lymphoblastic membranes (200‐10 000 × g pellets) was stimulated by peptides, in the presence of GTP, with the following order of potency: helodermin(1–35)‐NH2 > helo‐dermin(1–27)‐NH2 > helospectin = VIP = PHI.
Analysis of the stop codon context in plant nuclear genes
Authors: Angenon Geert , Van Montagu Marc , Depicker Ann
Published: July 2002

A region of 18 nucleotides surrounding the stop codon (the stop codon context) in 748 plant nuclear genes was analyzed. Non‐randomness was found both upstream and downstream from the stop codon, suggesting that these sequences may help in ensuring efficient termination of translation. The UAG amber codon is the least‐used stop codon and the bias in the nucleotide distribution 5' and 3' to the stop codon was more pronounced for the amber codon than for the other stop codons. This might indicate that the codon context affects termination more at UAG than at UGA or UAA stop codons.
Ribosomal localization of the mRNA in the 30S initiation complex as revealed by UV crosslinking
Authors: Brandt Roland , Gualerzi Claudio O.
Published: July 2002

Translation initiation complexes consisting of 30S ribosomal subunits, 32P‐labelled mRNA (002 mRNA), fMet‐tRNA and the three initiation factors were subjected to UV‐crosslinking to determine the protein and rRNA neighbors of the bound mRNA by immunochemical methods and by nucleic acid hybridization techniques. The mRNA was found to be crosslinked to a specific region of the 16S rRNA spanning from nucleotide 418 to 615 and to ribosomal proteins S1 and S21 (the main targets), S3, S10, S12 and S14; a low level of crosslinking was also detected with S2, S7, S13, S18 and S19.
Differential inhibition of fungal oxidosqualene cyclase by 6E and 6Z isomers of 2,3‐epoxy‐10‐aza‐10,11‐dihydrosqualene
Authors: Balliano Gianni , Milla Paola , Ceruti Maurizio , Viola Franca , Carrano Lùcia , Cattel Luigi
Published: July 2002

Inhibitory properties of 6E (compound 1) and 6Z (compound 2) isomers of 2,3‐epoxy‐10‐aza‐10,11‐dihydrosqualene against oxidosqualene‐lanosterol cyclase were assayed on microsomes and whole cells of Saccharomyces cerevisiae and Candida albicans. Only the 6E isomer (compound 1), bearing a correct substrate‐like configuration, strongly inhibited the enzyme both in microsomes and cell cultures. The difference between compounds 1 and 2 (which had an unfavorable geometry) was especially evident when measuring [14C]acetate incorporation into non‐saponifiable lipids extracted from treated cells. While isomer Z was totally ineffective at up to 30 μM, in cells treated with 5 μM isomer E, labelled oxidosqualene, the level of which was negligible in the control, rose to over 60% of the non‐saponifiable lipids.
Plasminogen‐binding protein associated with the plasma membrane of cultured embryonic rat neocortical neurons
Authors: Nakajima Kazuyuki , Nagata Koichi , Hamanoue Makoto , Takemoto Nagisa , Shimada Akinori , Kohsaka Shinichi
Published: July 2002

To investigate the receptor‐like molecule(s) for plasminogen (PGn) on the neuronal surface, the properties of binding of PGn to the plasma membrane of cultured embryonic rat neocortical neurons were investigated. [125I]PGn was found to specifically bind to the plasma membrane depending on the incubation temperature and time. The binding was also affected strongly by ionic strength and slightly by Ca2+. Furthermore, ligand blotting analysis revealed that [125I]PGn binds to a major protein with an apparent molecular weight of 45 kDa among plasma membrane proteins. These results suggest that the 45‐kDa protein is a PGn receptor‐like molecule on the neuronal surface.
Cephalopod alcohol dehydrogenase: purification and enzymatic characterization
Authors: Fernández M.Rosario , Jörnvall Hans , Moreno Alberto , Kaiser Rudolf , Parés Xavier
Published: July 2002

Octopus, squid and cuttle‐fish organs were examined for alcohol dehydrogenase activity. Only one form was detectable, with properties typical of mammalian class III alcohol dehydrogenase. The corresponding protein was purified from octopus and enzymatically characterized. Ionexchange and affinity chromatography produced a pure protein in excellent yield (73%) after 1600‐fold purification. Enzymatic parameters with several substrates were similar to those for the human class III alcohol dehydrogenase, demonstrating a largely conserved function of the enzyme through wide lines of divergence covering vertebrates, cephalopods and bacteria. The results establish the universal occurrence of class III alcohol dehydrogenase and its strictly conserved functional properties in separate living forms. The absence of other alcohol dehydrogenases in cephalopods is compatible with the emergence of the ethanol‐active class I type at a later stage, in lineages leading to vertebrates.
Alzheimer paired helical filaments Restoration of the biological activity by dephosphorylation
Authors: Iqbal Khalid , Zaidi Tanweer , Bancher Christian , Grundke-Iqbal Inge
Published: July 2002

In a normal mature neuron, microtubule associated protein tau promotes the assembly of tubulin into microtubules and maintains the structure of microtubules. In Alzheimer disease brain, tau is abnormally hyperphosphorylated and is the major protein subunit of paired helical filaments PHF). In the present study, the biological activity of tau in PHF and the effect of dephosphorylation on this activity were examined. PHF were isolated from Alzheimer disease brains and tau from the untreated or alkaline phosphatase‐treated PHF was extracted by ultrasonication in microtubule assembly buffer. Tubulin was isolated by phosphocellulose chromatography of three cycled microtubules from bovine brain. PHF‐tau did not promote assembly of bovine tubulin into microtubules whereas tau from the dephosphorylated PHF produced a robust microtubule assembly. These studies suggest (i) that in Alzheimer disease tau in PHF is functionally inactive because of abnormal phosphorylation and (ii) that the abnormally phosphorylated site(s) in PHF that inactivates PHF‐tau is accessible to enzymatic dephosphorylation in vitro.
Basepairing with 18S ribosomal RNA in internal initiation of translation
Authors: Scheper Gert C. , Voorma Harry O. , Thomas Adri A.M.
Published: July 2002

In concert with the translation initiation factors ‘trans‐acting’ factors function specifically during internal initiation on picornaviral mRNAs. Of these trans‐acting factors, two have been identified as the La‐protein and the polypyrimidine tract binding protein. Within the internal ribosomal entry site on the viral RNA, sequences are present that direct the ribosome to the initiation codon. We suggest that selection of the correct AUG initiation codon occurs through basepairing with a part of 18S ribosomal RNA.
Specific receptors for adrenomedullin in cultured rat vascular smooth muscle cells
Authors: Eguchi Satoru , Hirata Yukio , Kano Hiroaki , Sato Kyoko , Watanabe Yukihiko , Watanabe Takushi X. , Nakajima Kiichiro , Sakakibara Shumpei , Marumo Fumiaki
Published: July 2002

The effects of synthetic rat adrenomedullin (rAM), a novel vasorelaxant peptide originally isolated from human pheochromocytoma, on receptor binding and cAMP generation were studied in cultured rat vascular smooth muscle cells (VSMC). A binding study using [125I]rAM revealed the presence of a single class of high‐affinity (K
d1.3 × 10−8 M) binding sites for rAM in VSMC. The apparent K
i of rat calcitonin gene‐related peptide (rCGRP) was 3 × 10−7 M. Affinity labeling of VSMC membranes with [125I]rAM revealed two distinct labeled bands with apparent molecular weights of 120 and 70 kDa, both of which were abolished by excess unlabeled rAM or rCGRP. rAM stimulated cAMP formation with an approximate EC50 of 10−8 M, the effect of which was additive with isoproterenol, but not with rCGRP. The rAM‐induced cAMP response was unaffected by propranalol, indomethacin, or quinaerine, but inhibited by a CGRP receptor antagonist, human CGRP[8–37]. These data suggest that VSMC possesses specific AM receptors functionally coupled to adenylate cyclase with which CGRP interacts.
Wortmannin, a specific inhibitor of phosphatidylinositol‐3 kinase, blocks osteoclastic bone resorption
Authors: Nakamura Ichiro , Takahashi Naoyuki , Sasaki Takahisa , Tanaka Sakae , Udagawa Nobuyuki , Murakami Hiroshi , Kimura Koutaro , Kabuyama Yukihito , Kurokawa Takahide , Suda Tatsuo , Fukui Yasuhisa
Published: July 2002

The biological role of phosphatidylinositol (PI)‐3 kinase was examined in osteoclast‐like multinucleated cells (OCLs) formed in co‐cultures of mouse osteoblastic cells and bone marrow cells. The expression of PI‐3 kinase in OCLs was confirmed by Western blot analysis. Wortmannin (WT), a specific inhibitor of PI‐3 kinase, inhibited PI‐3 kinase activity in OCLs both in vitro and in vivo. WT also inhibited pit‐forming activity on dentine slices and disrupted a ringed structure of F‐actin‐containing dots (an actin ring) in OCLs in a dose‐dependent manner. The inhibitory profiles of WT for pit and actin ring formation were similar to that for PI‐3 kinase activity in OCLs. Electron microscopic analysis revealed that OCLs treated with WT did not form ruffled borders. Instead, numerous electron lucent vacuoles of differing sizes were found throughout the cytoplasm. These results suggest that PI‐3 kinase is important in osteoclastic bone resorption.
In vivo treatment of Heymann's Nephritis using a cytotoxic protein‐toxin conjugate
Authors: Misquith Sandra , Surolia Avadhesha
Published: July 2002

In this study we investigated the possibility of treating Heymann's Nephritis (HN) by destroying antibody producing cells by targetting a toxin, gelonin ‐ conjugated to gp330, the renal brush border antigen. HN was induced in rats by immunizing them with purified gp330. The gelonin‐gp330 conjugate was administered 12 days after the antigenic challenge. Serum was screened for circulating antibodies. Proteinurea was estimated. The gp330‐gelonin conjugate‐treated animals had a circulating antibody titre in the serum much lower than that of diseased (untreated) animals. Proteinurea seen in diseased animals was not observed in treated animals. This work suggests the possibility of using a toxin‐antigen conjugate for immunomodulating antibody mediated autoimmune renal disease.
Topological organization of subunits VII and VIII in the ubiquinolcytochrome c oxidoreductase of Saccharomyces cerevisiae
Authors: Boumans Hans , Berden Jan A. , Grivell Leslie A.
Published: July 2002

To determine the topology of subunit VIII of the yeast ubiquinol‐cytochrome c oxidoreductase in the mitochondrial inner membrane, an epitope has been introduced in the N‐terminal half of this protein. Previous topology studies had shown that at least the C‐terminus faces the intermembrane space [Hemrika and Berden (1990) Eur. J. Biochem. 192, 761–765]. Based on sensitivity of the protein to proteinase K digestion we now suggest that the N‐terminus of subunit VIII is similarly oriented, implying that this subunit does not span the membrane. Despite this, however, subunit VIII cannot be extracted from the membrane even after treatment with 0.1 M Na2CO3 at pH 11.5, showing that the protein is integrally embedded in the membrane. A similar behaviour was displayed by another low molecular weight protein of the complex, subunit VII, which faces the matrix side. A model for the topology of these subunits in the membrane is discussed with respect to the structure of the complex and their involvement in quinone binding.
Development of vestibular and auditory function: Effects of hypothyroidism and thyroxine replacement therapy on nystagmus and auditory evoked potentials in the pigmented rat
Authors: Meza Graciela , Acuña Dora , Escobar Carolina
Published: July 2002

The functional development of semicircular canals and some brainstem structures of the auditory system was followed in parallel with time in control and propylthiouracyl‐induced hypothyroid pigmented rats by respective recording of postrotatory nystagmus response and auditory evoked brainstem potentials, with the aim of discovering the timing of permanent alterations of these responses in congenital hypothyroidism. A group of hypothyroid rats which underwent thyroxine‐replacement therapy from postnatal day 12 onward was also included in our studies to corroborate the involvement of thyroid hormones in these effects. Postrotatory nystagmus and auditory evoked responses were absent in congenital hypothyroid rats. In the thyroxine‐replaced group postrotatory nystagmus values showed no differences from the control group from postnatal day 28 onward. Auditory evoked potentials in thyroxine‐replaced animals could not be elicited at 30 dB, but by increasing the intensity of stimulus to 70 dB, values of latencies of the four waves composing the response were indistinguishable from controls from postnatal day 39 and thereafter. These results show that hypothyroidism affects both semicircular canal and auditory function, the latter more severely than the former, but that these effects can be prevented when thyroxine replacement treatment is started in early stages of postnatal development.
Outpatient treatment of acute psychotic episode with neuroleptic infusions: a retrospective study
Authors: Bonsack CL , Philippoz R
Published: July 2002

In this retrospective pragmatic study, we define the necessary conditions that allow outpatient low dose intravenous neuroleptization, when hospitalization should otherwise be required. Intravenous neuroleptization is infrequently used in the outpatient treatment of acute psychotic decompensation. Rapid tranquilization with high dosage neuroleptics is controversial, and has a high risk of side effects. The indications for and potential advantages of this method in the perspective of a long-term ambulatory treatment are discussed by comparing a group of outpatients treated with infusions to a group of hospitalized patients. The method offers a satisfactory alternative to hospitalization for subjects who are not in imminent danger (current GAF rating between 20 and 40) and whose normal functioning is good (past year GAF rating = 70). Previous repeated hospitalizations favor the choice of hospitalization over infusion. Its potential advantages are the rapid evolution of the condition, with controlled regression but without psychosocial withdrawal, and an improvement in the patient's attitude towards treatment.
Rapid internal dynamics of BPTI is insensitive to pressure
Authors: Sareth Sina , Li Hua , Yamada Hiroaki , Woodward Clare K. , Akasaka Kazuyuki
Published: July 2002

Pressure effects on the backbone dynamics of a native basic pancreatic trypsin inhibitor (BPTI) have been measured by 15N spin relaxation and chemical shifts at 30 and 2000 bar. The experiments utilized the on‐line variable pressure cell nuclear magnetic resonance system on 15N‐uniformly labeled BPTI at a proton frequency of 750.13 MHz at 36°C. Longitudinal (R
1) and transverse (R
2) 15N relaxation times and (1H)–15N nuclear Overhauser effects were measured for 41 protonated backbone nitrogens at both pressures. The model free analysis of the internal dynamics gave order parameters for individual H–N vectors at both pressures. The results indicate that rapid internal dynamics in the ps–ns range for the polypeptide backbone is not significantly affected by pressure in the range between 30 bar and 2 kbar. The result is consistent with the linear pressure dependence of 1H and 15N chemical shifts of BPTI, which suggests that local compressibilities and amplitudes of associated conformational fluctuation are nearly invariant in the same pressure range. Overall, we conclude that at 2 kbar BPTI remains within the same native ensemble as at 1 bar, with a small shift of population from that at 1 bar.
Membrane anchoring and surface distribution of glycohydrolases of human erythrocyte membranes
Authors: Goi Giancarlo , Bairati Chiara , Massaccesi Luca , Lovagnini Augusto , Lombardo Adriana , Tettamanti Guido
Published: July 2002

The membrane anchoring of the following glycohydrolases of human erythrocyte plasma membranes was investigated: α‐ and β‐D‐glucosidase, α‐ and β‐D‐galactosidase, β‐D‐glucuronidase, N‐acetyl‐β‐D‐glucosaminidase, α‐D‐mannosidase, and α‐L‐fucosidase. Optimized fluorimetric methods for the assay of these enzymes were set up. Treatment of the ghost preparation with 1.0 mol/l (optimal concentration) NaCl caused release ranging from 4.2% of α‐D‐glucosidase to 70% of β‐D‐galactosidase; treatment with 0.4% (optimal concentration) Triton X‐100 liberated 5.1% of β‐D‐galactosidase to 89% of α‐D‐glucosidase; treatment with 1.75% (optimal concentration) octylglucoside yielded solubilization from 6.3% of β‐D‐galactosidase to 85% of α‐D‐glucosidase. Treatment with phosphoinositide‐specific phospholipase C caused no liberation of any of the studied glycohydrolases. These results are consistent with the notion that the above glycohydrolases are differently anchored or associated with the erythrocyte plasma membrane, and provide the methodological basis for inspecting the occurrence of these enzymes in different membrane microdomains.
Density‐dependent induction of TNF‐α release from human monocytes by immobilized P‐selectin
Authors: Koike Junzo , Nagata Kisaburo , Kudo Sanae , Tsuji Tsutomu , Irimura Tatsuro
Published: July 2002

P‐selectin purified from human platelets, when immobilized on a solid surface, induced monocytes to release tumor necrosis factor‐α (TNF‐α). The induction of TNF‐α release was dependent on the concentration of P‐selectin used for the immobilization, and the maximal stimulation was observed when the plate was coated with 0.3 μg/ml of P‐selectin. Use of either a higher or a lower concentration of P‐selectin for the plate‐coating was found to elicit less TNF‐α release, although the higher concentration of P‐selectin caused a stronger adhesion of HL‐60 leukemic cells. The expression of mRNA for TNF‐α roughly paralleled the TNF‐α secretion, as assessed by RT‐PCR. These results indicate that monocytes are activated by immobilized P‐selectin in a density‐dependent manner.
Efficient GFP mutations profoundly affect mRNA transcription and translation rates
Authors: Sacchetti Andrea , El Sewedy Tarek , Nasr Ashraf F , Alberti Saverio
Published: July 2002

Green fluorescent protein (GFP) variants with higher expression efficiencies have been generated by mutagenesis. Favorable mutations often improve the folding of GFP. However, an effect on protein folding fails to explain the efficiency of several other GFP mutations. In this work, we demonstrate that mutations of the GFP open reading frame and untranslated regions profoundly affect mRNA transcription and translation efficiencies. The removal of the GFP 5′ untranslated region halves the transcription rate of the GFP gene, but hugely improves its translation rate. Mutations of the GFP open reading frame or the addition of peptide sequences differentially reduce the GFP mRNA transcription rate, translation efficiency and protein stability. These previously unrecognized effects are demonstrated to be critical to the efficiency of GFP mutants. These findings indicate the feasibility of generating more efficient GFP variants, with optimized mRNA transcription and translation in eukaryotic cells.
Is leptin an insulin counter‐regulatory hormone?
Authors: Remesar X , Rafecas I , Fernández-López J.A , Alemany M
Published: July 2002

Leptin, the product of the ob gene, controls appetite through the hypothalamus and may affect many other tissues because of the widespread distribution of its receptors. Leptin is synthesized by white adipose tissue (WAT) under conditions of high energy availability and insulin stimulus. Glucocorticoids enhance this synthesis and catecholamines hamper leptin production. Leptin diminishes insulin secretion by the pancreatic beta cells and induces insulin resistance. In fact leptin hampers insulin action on WAT itself in a negative feedback loop. The evidence acquired in studies on diabetics, starvation, refeeding and insulin and glucose clamps supports this interpretation, which may also explain part of the difficulties encountered by the current postulate that links leptin to WAT mass size signalling to the brain. Leptin may be, essentially, a counter‐regulatory hormone limiting the insulin drive to store energy in the form of fat, its effects reaching from a decrease in food intake to lower insulin secretion and increased resistance to insulin and lower glucose uptake and fat synthesis by WAT.
Inhibitors of phosphoprotein phosphatases 1 and 2A cause activation of a 53 kDa protein kinase accompanying the apoptotic response of breast cancer cells
Authors: Rossini Gian Paolo , Pinna Cristina , Viviani Romano
Published: July 2002

Treatment of MCF‐7 breast cancer cells with 50 nM okadaic acid triggers an apoptotic response which is accompanied by a 7‐fold increase in the activity of a protein kinase with a relative molecular mass of 53 kDa. The activity of the kinase was stimulated by cell treatment with inhibitors of phosphoprotein phosphatase 1 and 2A, but not by stressing conditions. Okadaic acid‐induced stimulation of the 53 kDa protein kinase was not abolished by coincubation of cells with cycloheximide. We conclude that stimulation of the 53 kDa protein kinase by inhibitors of phosphoprotein phosphatases involves pre‐existing molecular components whose activity depends on the phosphorylation state of serine/threonine residues.
Phosphocholine and sphingosine‐1‐phosphate synergistically stimulate DNA synthesis by a MAP kinase‐dependent mechanism
Authors: Kiss Zoltan , Mukherjee Jagat J
Published: July 2002

We have previously shown that in NIH 3T3 fibroblasts phosphocholine (PCho) potentiates sphingosine‐1‐phosphate (S1P)‐induced mitogenesis. Here we report that PCho and S1P also synergistically stimulate DNA synthesis in mouse Swiss 3T3 fibroblasts and in mouse JB6 epidermal cells. The combined actions of PCho and S1P on DNA synthesis were associated with synergistic activation of the p42/p44 mitogen‐activated protein (MAP) kinases. Ethanolamine (50–100 μM) further enhanced the synergistic effects of PCho and S1P on DNA synthesis but not on MAP kinase activity. The results indicate that the synergistic mitogenic effects of PCho and S1P (i) are not restricted to NIH 3T3 fibroblasts, (ii) are predominantly mediated by the MAP kinase‐dependent signal transduction pathway, and (iii) are enhanced by ethanolamine via a MAP kinase‐independent mechanism.
Chinese hamster protein homologous to human putative protein kinase KIAA0204 is associated with nuclei, microtubules and centrosomes in CHO‐K1 cells
Authors: Zinovkina Lyudmila A. , Poltaraus Andrey B. , Solovyanova Olga B. , Nadezhdina Elena S.
Published: July 2002

Monoclonal antibody raised against a preparation of loach fish sperm centrosomes was used for screening of cDNA expressing library of Chinese hamster CHO‐K1 cells. Two positive clones appeared to encode 628 amino acid protein fragment that was 72% identical to human KIAA0204 protein, i.e. putative protein kinase. Polyclonal antibodies raised against products of cDNA expression in E. coli recognized 210‐kDa polypeptide in CHO‐K1 cells and immunostained nuclear speckles, centrosomes and microtubules in these cells. The 210‐kDa polypeptide (named MAK‐L) co‐sedimented with exogenous microtubules. Thus, one more protein kinase seems to be associated with the microtubule network in vertebrate cells.
Carbohydrate induced modulation of cell membrane VII. Binding of exogenous lectin increases osmofragility of erythrocytes
Authors: Pande Abhay H. , Sumati  , Hajela Namita , Hajela Krishnan
Published: July 2002

Due to their multivalent binding character, lectins when added exogenously will cross‐link membrane surface receptors leading to lateral molecular reorganizations in the plane of the bilayer. This study reports for the first time that agglutination of rabbit erythrocytes by lentil lectin and concanavalin A increases their osmofragility. Increase in osmofragility was detected by measuring the hemolysis of erythrocytes in hypotonic as well as in isotonic solutions. It was also found that agglutination per se does not increase osmofragility but the binding of legume lectin is essential since human Rh+ cells agglutinated by a monoclonal antibody do not exhibit hemolysis.
RNA aptamers that specifically bind to the Ras‐binding domain of Raf‐1
Authors: Kimoto Michiko , Sakamoto Kensaku , Shirouzu Mikako , Hirao Ichiro , Yokoyama Shigeyuki
Published: July 2002

RNA aptamers that bind to the Ras‐binding domain (RBD) of a proto‐oncogene product, Raf‐1, were isolated from a pool of random sequences using a glutathione S‐transferase‐fused RBD (GST‐RBD). The RNA molecules bind to the GST‐RBD, but not to GST, with dissociation constants of about 300 nM. In contrast, these RNA aptamers do not bind to the Ras‐binding domain of the RGL protein, which is also known to be activated by Ras. The aptamers actually compete with Ras for binding to the Raf‐1 RBD. The anti‐Raf‐1 aptamers may be used to specifically inhibit the Ras‐Raf interaction in the complicated signaling network in mammalian cells.
Early release and subsequent caspase‐mediated degradation of cytochrome c in apoptotic cerebellar granule cells
Authors: Bobba A. , Atlante A. , Giannattasio S. , Sgaramella G. , Calissano P. , Marra E.
Published: July 2002

Cytochrome c (cyt c) release was investigated in cerebellar granule cells used as an in vitro neuronal model of apoptosis. We have found that cyt c is released into the cytoplasm as an intact, functionally active protein, that this event occurs early, in the commitment phase of the apoptotic process, and that after accumulation, this protein is progressively degraded. Degradation, but not release, is fully blocked by benzyloxycarbonyl‐Val‐Ala‐Asp‐fluoromethylchetone (z‐VAD‐fmk). On the basis of previous findings obtained in the same neuronal population undergoing excitotoxic death, it is hypothesized that release of cyt c may be part of a cellular attempt to maintain production of ATP via cytochrome oxidase, which is reduced by cytosolic NADH in a cytochrome b
5‐soluble cyt c‐mediated fashion.
Sonographic features of trisomy 13 at midpregnancy
Authors: Tongsong T. , Sirichotiyakul S. , Wanapirak C. , Chanprapaph P.
Published: July 2002

Objectives: To evaluate the sonographic characteristics, at 16–22 weeks of gestation, of fetuses later diagnosed with trisomy 13. Methods: This descriptive analysis of a case series was conducted from June 1989 to May 2001 at the Department of Obstetrics and Gynecology, Faculty of Medicine, Chiang Mai University, Thailand. Women with abnormal sonographic findings at midpregnancy (16–22 weeks of gestation) underwent amniocentesis or cordocentesis for karyotyping, and the inclusion criterion was proven trisomy 13. Results: Indications for sonographic examination at midpregnancy were a genetic risk, large‐ or small‐for‐date fetus, and other suspected anomalies. Fifteen fetuses were later diagnosed with trisomy 13. In all of these cases there was at least one abnormal sonographic finding. In only one case did the fetus show no structural abnormality (at 17 weeks), but polyhydramnios and fetal growth restriction were observed. Common sonographic findings included holoprosencephaly with associated facial anomalies, and abnormal feet and/or hands, especially polydactyly. Non‐structural abnormal findings such as polyhydramnios or fetal growth restriction were seen in less than one third of the fetuses. Conclusions: Nearly all the fetuses with trisomy 13 had sonographic characteristics of abnormalities at midpregnancy although common findings had often not yet appeared or the findings were low‐sensitive. Detailed ultrasound at midpregnancy could effectively screen for further genetic testing pregnancies at risk for trisomy 13.
Improving recordkeeping for maternal mortality programs, Kumasi, Ghana
Authors: Danquah J.B , Appah E.K , Djan J.O , Ofori M , Essegbey I.T , Opoku S ,   (The Kumasi PMM Team)
Published: July 2002

Preliminary studies: Data on obstetric complications are the basis of monitoring maternal mortality interventions in the PMM Network. A review of recordkeeping procedures at 10 facilities in the study area revealed that information on obstetric complications was often inconsistent or missing. Some hospital records were not designed to collect such information at all. Interventions: In 1992, registers at facilities were revised to collect information on complications and time of treatment. Doctors, nurses, midwives and clerks were trained to record, compile and analyze data. Monitoring and supervisory mechanisms were also set up. Results: Recordkeeping has improved. Data collection and analysis have been regular and timely. Doctors have begun using the data for morning meetings. Nurses and midwives compile monthly summaries of data showing complications by type. Two other districts outside the research area have adopted the reporting system and it is possible that facilities in the whole region will follow suit. Costs: The cost of improving recordkeeping at the 10 healthcare facilities was approximately US $2543, with 85% coming from project funds. Conclusions: Existing recordkeeping systems can be modified to collect data necessary to monitor maternal mortality interventions. Staff training and monitoring visits are important to success. © 1997 International Federation of Gynecology and Obstetrics
Pharmacotherapy of dysthymia: review and new findings
Authors: Versiani M , Nardi AE , Figueira I
Published: July 2002

Open trials with tricyclics, classical monoamine oxidase inhibitors (MAOIs) or lithium in dysthymia yielded a response rate in 45% of subjects. A long-term treatment of dysthymia with 276 patients treated during 4 years with either moclobemide, tranylcypromine or a combination of amitryptiline plus chlordiazepoxide is described. After discontinuation there was a relapse rate of 89.1%. The controlled studies with tricyclics, MAOIs, reversible inhibitors of monoamine oxidase (RIMAs), specific serotonin reuptake inhibitor (SSRIs) or benzamides showed that drugs well-tolerated work better in dysthymia, due to the fact that the treatment must be long-term. Sertraline was studied vs placebo or imipramine in primary dysthymia. Moclobemide, imipramine and placebo were also studied in 315 patients. Mean doses were 650 mg/d of moclobemide and 203.2 mg/d of imipramine. Moclobemide and sertraline were both efficacious and well tolerated. In a long term treatment the clinician should assess the risk-benefit ratio. Dysthymic patients are very sensitive to unwanted effects and compliance is a serious issue.
Dividend initiations in reverse‐LBO firms
Authors: Kosedag Arman , Michayluk David
Published: July 2002

This article documents a lack of share price response to dividend initiation announcements by firms that recently completed a reverse‐LBO. We identify a group of firms that had recently completed an IPO and report that they experience a positive reaction to dividend initiation announcements as expected based on the prior dividend initiation literature. We rule out a size‐based explanation for the abnormal response in reverse‐LBOs and offer some potential explanations for our empirical findings.
DOCA‐salts induce heart failure in the guinea pig
Authors: Tiritilli Alberto
Published: July 2002

Heart failure (HF) is a common clinical problem confronting physicians and is often the final manifestation of many cardiovascular disorders. Despite recent advances in the pharmacological management of HF, it remains a highly lethal and disabling disorder. A number of animal models have been developed to study both the pathophysiology of HF and new therapeutic approaches to this complex syndrome. Only through an improved understanding of the basic biology of the early stages of the syndrome can HF be prevented or at least anticipated. With this in view, we have developed an easily realisable and inexpensive model in the guinea pig, which presents numerous structural, metabolic and biochemical similarities compared with the human heart. Thirty guinea pigs, aged 5 weeks and weighing 300 g were used. After anaesthesia, left nephrectomy was performed. After 1 week the guinea pigs were divided into: (a) control group (n ‐ 15), which received an injection of vehicle as well as tap water for 10 weeks; (b) DOCA‐salts group (n ‐ 15), where the animals were treated with an IM injection of 10 mg DOCA 5 days a week for 10 weeks and with drinking water containing 9 g/l−1 NaCl and 2 g/l−1 KCl. Our results demonstrate that the administration of DOCA‐salts to guinea pigs for 10 weeks caused a significant increase in blood pressure (BP + 30%) associated with left ventricular hypertrophy (LVH), evaluated by LV weight (+37%), LV wall (+36%), by the ratio LV weight/Body weight (+23%) and by an increase in LV volume (+51%). Concerning HF, the latter was clinically evident through an increase in body weight, heart rate and dyspnoea. Indeed, guinea pigs presented pleural and/or pericardial effusion often associated with ascite. This model, which combines pressure and volume overload, results in a slow evolution towards HF. This allows a better understanding of the mechanisms in early LV remodelling which has the potential to develop into HF. Some recent studies have emphasised the value of using guinea pigs. The guinea pig heart muscle presents two major regulatory mechanisms of contractility that are closer to those found in humans, the isomyosin pattern which is predominantly V3 and the phenomenon of Ca2+‐induced Ca2+‐release from the sarcoplasmic reticulum. The DOCA‐salts model in the guinea pig is an easy surgical procedure with high post‐operative survival, which causes an increase in arterial BP, LVH associated with HF. This model is a useful tool for studying some of the basic mechanisms of cardiovascular diseases.
Altering element interactivity and variability in example‐practice sequences to enhance learning to write Chinese characters
Authors: Lu Jia , Kalyuga Slava , Sweller John
Published: April 2020

The isolated elements and variability effects of cognitive load theory were used to alter the element interactivity of Chinese characters when instructing novice learners (72 overseas students at a Chinese university) in writing characters using worked examples‐practice procedures. A group of characters with more than eight strokes was disassembled into three individual components. Students were required to trace examples and practice writing of either similar sequences of isolated components followed by integrated components of full characters (Isolated–Integrated sequences); variable sequences of isolated followed by integrated components (Variability–Integrated sequences); or conventional Integrated–Integrated sequences of full characters. It was hypothesized that the fully integrated stroke‐movements form of example‐practice sequences would result in less learning due to a greater cognitive load. The results demonstrated that the participants in both the Isolated–Integrated and Variable–Integrated groups performed significantly better than the Integrated–Integrated group with the Variable–Integrated group outperforming the Isolated–Integrated group.
Variability in Interpretation of Magnetic Resonance Imaging of the Pediatric Sacroiliac Joint
Authors: Weiss Pamela F. , Brandon Timothy G. , Bohnsack John , Heshin‐Bekenstein Merav , Francavilla Michael L. , Jaremko Jacob L. , Liao Lester , McHugh Anne , Oberle Edward J. , Rumsey Dax , Srinivasalu Hemalatha , Stoll Matthew L. , Chauvin Nancy A.
Published: April 2020

Objective

Magnetic resonance imaging (MRI) is pivotal in the assessment of early sacroiliitis in children. We aimed to evaluate the agreement between local radiology reports and central imaging reviewers for active inflammation and structural damage at the sacroiliac (SI) joints.

Methods

Eight hospitals each contributed up to 20 cases of consecutively imaged children and adolescents with juvenile idiopathic arthritis and suspected sacroiliitis. Studies were independently reviewed by 3 experienced musculoskeletal pediatric radiologists. Local assessments of global impression and lesions were coded from the local radiology reports by 2 study team members. Test properties of local reports were calculated using the central imaging team’s majority as the reference standard.

Results

For 120 evaluable subjects, the median age was 14 years, half of the cases were male, and median disease duration at the time of imaging was 0.8 years (interquartile range 0–2). Sensitivity of local reports for inflammation was high, 93.5% (95% confidence interval [95% CI] 78.6–99.2), and specificity was moderate, 69.7% (95% CI 59.0–79.0), but positive predictive value (PPV) was low, 51.8% (95% CI 38.0–65.3). Twenty‐seven cases (23%) had active inflammation reported locally but rated normal at the central reading, 19 (70%) with subsequent medication changes. The sensitivity of local reports detecting structural damage was low, 45.7% (95% CI 28.8–63.4), and specificity was high, 88.2% (95% CI 79.4–94.2); PPV was low, 61.5% (95% CI 40.6–79.8).

Conclusion

Substantial variation exists in the interpretation of inflammatory and structural lesions at the SI joints in children. To reliably identify pathology, additional training in the MRI appearance of the maturing SI joint is greatly needed.
Cover cropping increased phosphorus stocks in surface sandy Ultisols under long‐term conservation and conventional tillage
Authors: Ye Rongzhong , Parajuli Binaya , Ducey Thomas F. , Novak Jeffrey M. , Bauer Phil J. , Szogi Ariel A.
Published: April 2020

Low use‐efficiency and high environmental significance of phosphorus (P) requires a better understanding of its stocks and behavior in soils. We investigated P fractions in sandy Coastal Plain Ultisols, where long‐term conservation agriculture driven by conservation tillage and residue return for ∼40 yr and the integration of cover cropping for 4 yr has been demonstrated to improve soil organic matter. Soils were collected from fields at 0‐ to 5‐ and 5‐ to 15‐cm depths to study the effects of tillage (conservation vs. conventional) and cover crop (with vs. without) on soil stocks of various chemically defined P pools and the phosphatase potential activities. Conservation tillage increased KCl‐extractable inorganic P (KCl‐Pi) stocks in top soils (0–5 cm) when compared to conventional tillage, but had no effects on other pools at both soil depths. Cover cropping caused significant accumulations of NaOH‐extractable organic P (NaOH‐Po) in top soils (0–5 cm). Nonetheless, neither conservation tillage nor cover crop changed the contributions of the chemically defined pools to soil total P with NaOH‐Po dominating at both soil depths followed by NaOH‐extractable inorganic P (NaOH‐Pi) and HCl‐extractable Pi (HCl‐Pi). Conservation tillage increased phosphatase potential activities by 128% in the 0‐ to 5‐cm soils, whereas no cover crop effects were observed. Conservation tillage improved P availability potentially through its effects on microbial activities, whereas cover cropping increased P stocks and availability by promoting Po accumulations.
A composite measure of healthy lifestyle: A study from 38 countries and regions from Europe and North America, from the Health Behavior in School‐Aged Children survey
Authors: Marques Adilson , Bordado Joana , Tesler Riki , Demetriou Yolanda , Sturm David J. , de Matos Margarida Gaspar
Published: April 2020

Introduction

This study aimed to create a composite measure of a healthy lifestyle for adolescents, and analyze its relationship to sociodemographic factors.

Methods

Data were from the Health Behavior in School‐Aged Children 2014 International survey. Participants were 167 021 adolescents (48.2% boys and 51.8 girls), aged 10‐16 years, from 38 countries.

Results

Five healthy behaviors used in this study included engaging in ≥60 minutes of physical activity every day, daily consumption of fruit and vegetables, spending <2 hours daily immersed in screen‐based behaviors, and abstinence from alcohol as well as from tobacco products. Only 1.9% (95% CI: 1.4%, 2.3%) of adolescents had a healthy lifestyle, achieving all five healthy behaviors. In contrast, 4.2% (95% CI: 3.7%, 4.6%) reported none of the healthy behaviors.

Conclusions

Despite the benefits of engaging in physical activity, engaging in low levels of screen‐based activity, regular consumption of fruits and vegetables, and abstaining from alcohol and cigarettes, only 2% of adolescents could be classified as having a healthy lifestyle.