bluuebunny/crossref_metadata_2025 huggingface dataset:
A novel peridinin—chlorophyll a protein (PCP) from the marine dinoflagellate Alexandrium cohorticula: A high pigment content and plural spectral forms of peridinin and chlorophyll a
Authors: Ogata Takehiko , Kodama Masaaki , Nomura Setsuzo , Kobayashi Masayuki , Nozawa Tsunenori , Katoh Tetzuya , Mimuro Mamoru
Published: July 2002

A new type of peridinin—chlorophyll a protein (PCP) was isolated from the marine dinoflagellate Alexandrium cohorticula. Unlike previous studies, PCP was obtained as a single component in the presence of a protease inhibitor. The monomer had a molecular mass of 37 kDa with 12 peridinin molecules associated with 2 chl a molecules. This pigment content was much higher than that reported previously. We observed a partial amino acid sequence of the N‐terminus that is novel among photosynthetic pigment—protein complexes. Magnetic circular dichroism clearly indicated that chl a in PCP had monomeric features. Multiple spectral components were suggested for both chl a and peridinin. Based on the high pigment content, the optical properties were compared with those for a reported PCP containing 4 chl a and 1 peridinin.
Isolation of an ion channel gene from Arabidopsis thaliana using the H5 signature sequence from voltage‐dependent K+ channels
Authors: Ketchum Karen A. , Slayman Carolyn W.
Published: July 2002

A degenerate oligonucleotide corresponding to the K+ channel signature sequence (TMTTVGYGD) was used to isolate the genomic and cDNA forms of a new channel gene, AKT3, from Arabidopsis thaliana. The deduced protein sequence has a predicted membrane topography similar to Shaker‐like K+ channels. Three distinct modules comprise the carboxyl‐terminal half: a nucleotide‐binding motif, an ankyrin repeat domain, and a polyglutamate track. Xenopus oocytes injected with cRNA exhibited an inward‐rectifying K+ current, demonstrating that the AKT3 polypeptide is a functional transport protein. Two other Arabidopsis K+ transporters (AKT1 and KAT1) share 60% homology with AKT3; together these proteins constitute a family of plant inward‐rectifying K+ channels.
Transvaginal trucut biopsy in patients with abdominopelvic mass
Authors: Sheth S.S. , Angirish J.
Published: July 2002

Objective: To perform an abdominopelvic or pelvic mass biopsy as an office procedure, as an alternative to laparoscopic biopsy. Methods: A transvaginal trucut biopsy using a disposable biopsy needle was carried out in 5 cases. Results: The biopsy gave a clear diagnosis of ovarian cancer in 2 cases, non‐tubercular inflammation in 2 cases and tuberculosis in 1 case. Conclusions: In the developing world, transvaginal trucut biopsy is an important method for diagnosis.
Stimulation of glutamine synthetase activity by excitatory amino acids in astrocyte cultures derived from aged mouse cerebral hemispheres may be associated with non‐n‐methyl‐d‐aspartate receptor activation
Authors: Fleischer‐Lambropoulos Eleni , Kazazoglou Theodosia , Geladopoulos Taxiarchis , Kentroti Susan , Stefanis Costas , Vernadakis Antonia
Published: July 2002

We have been using glial cells derived from aged mouse cerebral hemispheres (MACH) at several passages to study the responsiveness of astrocytes to microenvironmental signals in culture. In the present study, we examined the effects of excitatory amino acids on the activity of glutamine synthetase, a marker for astrocytes. MACH glia cell passages 25 to 29 were used. Culture groups were Dulbecco's modified Eagle's medium +10% fetal bovine serum (control); glutamate 100 μM; γ‐amino‐3‐hydroxy‐5‐methyl isoxazole‐4‐propionic acid (AMPA) 50 μM; kainic acid 10 μM; N‐methyl‐d‐aspartate (NMDA) 10 μM. In all treated groups glutamine synthetase activity was significantly higher than in controls. We speculate that this increase represents an enhanced differentiation of immature astrocytes. In a second series, we examined the effects of glutamate receptor antagonists on glutamine synthetase activity as follows. MACH cultures were treated with glutamate 100 μM in combination with either L(+)‐2‐amino‐3‐phosphonopropionic acid (L‐AP3; 50 μM); D(−)‐2‐amino‐5‐phosphonopentanoic acid (D‐AP5; 50 μM) or 6,7‐dinitroquinoxaline‐2,3‐dione (DNQX; 50 μM). The increase in GS activity produced by glutamate was exhibited by the non‐selective NMDA receptor antagonist, DNQX, but not by the metabotropic receptor antagonist, L‐AP3 or a selective NMDA receptor antagonist, D‐AP5. We also found that in cultures treated with glutamate, a number of astrocytes resembled “reactive astrocytes” morphologically. These astrocytes were absent in cultures treated with glutamate + DNQX. The findings provide supportive evidence that astrocytes from aged mouse cerebral hemispheres respond to excitatory amino acids and that this response is mediated by non‐NMDA receptor activation.
Nuclear wavepacket motion producing a reversible charge separation in bacterial reaction centers
Authors: Yakovlev A.G. , Shkuropatov A.Y. , Shuvalov V.A.
Published: July 2002

The excitation of bacterial reaction centers (RCs) at 870 nm by 30 fs pulses induces the nuclear wavepacket motions on the potential energy surface of the primary electron donor excited state P*, which lead to the fs oscillations in stimulated emission from P* [M.H. Vos, M.R. Jones, C.N. Hunter, J. Breton, J.‐C. Lambry and J.‐L. Martin (1994) Biochemistry 33, 6750–6757] and in QY absorption band of the primary electron acceptor, bacteriochlorophyll monomer BA [A.M. Streltsov, S.I.E. Vulto, A.Y. Shkuropatov, A.J. Hoff, T.J. Aartsma and V.A. Shuvalov (1998) J. Phys. Chem. B 102, 7293–7298] with a set of fundamental frequencies in the range of 10–300 cm−1. We have found that in pheophytin‐modified RCs, the fs oscillations with frequency around 130 cm−1 observed in the P*‐stimulated emission as well as in the BA absorption band at 800 nm are accompanied by remarkable and reversible formation of the 1020 nm absorption band which is characteristic of the radical anion band of bacteriochlorophyll monomer BA
−. These results are discussed in terms of a reversible electron transfer between P* and BA induced by a motion of the wavepacket near the intersection of potential energy surfaces of P* and P+BA
−, when a maximal value of the Franck–Condon factor is created.
The proteasome inhibitor MG132 promotes accumulation of the steroidogenic acute regulatory protein (StAR) and steroidogenesis
Authors: Tajima Kimihisa , Babich Svetlana , Yoshida Yoshio , Dantes Ada , Strauss Jerome F. , Amsterdam Abraham
Published: July 2002

StAR, a protein synthesized in the cytoplasm and subsequently imported into mitochondria, regulates the rate‐determining step in steroidogenesis, the transport of cholesterol from the outer to the inner mitochondrial membrane. The active form of StAR is the 37 kDa pre‐protein, which has a short half‐life. To determine whether proteasomes participate in the turnover of StAR, we incubated primary cultures of preovulatory rat granulosa cells and immortalized human granulosa cells in the presence of MG132, a specific inhibitor to proteasome catalysis. This treatment caused accumulation of StAR in unstimulated cells. Moreover, incubation of the cells with MG132 in the presence of forskolin (FK), luteinizing hormone/chorionic gonadotropin or follicular stimulating hormone augmented the accumulation of both the 37 kDa cytoplasmic protein and the 30 kDa mature mitochondrial protein, compared to cells incubated with FK or the gonadotropic hormones alone. Concomitantly, progesterone production was enhanced. In contrast no elevation in the 37 kDa StAR intracellular levels or progesterone production was observed following incubation of the cells with the cysteine protease inhibitor E‐64. The increase of the 37 kDa StAR protein was evident after 15 min and 30 min of incubation with MG132 (143% and 187% of control values, respectively) with no significant elevation of the 30 kDa protein. Accumulation of the intermediate mitochondrial 32 kDa protein was evident after 1–2 h and the accumulation of the 30 kDa protein was evident only after 4 h of incubation with MG132. In contrast, no elevation in adrenodoxin, a component of the cytochrome P450scc enzyme system, was found. These data suggest that StAR protein is either directly or indirectly degraded by the proteasome which may explain, in part, its short half‐life. Moreover, it seems that the cytosolic 37 kDa protein, which is responsible for the steroidogenic activity of StAR, is the primary proteasomal substrate and that the inhibition of its degradation by MG132 causes the up‐regulation of progesterone production.
Testosterone inhibits osteoclast formation stimulated by parathyroid hormone through androgen receptor
Authors: Chen Qingxiang , Kaji Hiroshi , Sugimoto Toshitsugu , Chihara Kazuo
Published: July 2002

Androgens play an important role in the regulation of bone metabolism in animals and humans. The present study was performed to investigate whether androgens would affect osteoclast formation stimulated by parathyroid hormone (PTH) in mouse bone cell cultures and its mechanism. Testosterone as well as α‐dihydrotestosterone (DHT) concentration‐dependently inhibited osteoclast formation induced by PTH‐(1‐34). 10−8 M ICI 182780, an estrogen receptor inhibitor, did not affect PTH‐induced osteoclast formation antagonized by 10−8 M testosterone, although it completely antagonized the effects of 10−8 M 17β‐estradiol. Moreover, 3 μM 4‐androsten‐4‐ol‐3,17‐dione, an aromatase inhibitor, did not affect PTH‐induced osteoclast formation antagonized by testosterone. Hydroxyflutamide, an androgen receptor antagonist, concentration‐dependently antagonized the inhibitory effects of testosterone as well as DHT on PTH‐stimulated osteoclast formation. In conclusion, the present study first demonstrated that testosterone inhibited osteoclast formation stimulated by PTH through the androgen receptor, but not through the production of intrinsic estrogen in mouse bone cell cultures.
Overexpression of Bcl‐2 suppresses the calcium activation of a mitochondrial megachannel
Authors: Murphy Robert C. , Schneider Erasmus , Kinnally Kathleen W.
Published: July 2002

The molecular mechanism(s) by which Bcl‐2 regulates apoptosis is poorly understood. Bcl‐2 suppresses apoptosis by inhibiting calcium activation of the permeability transition of mitochondria. In this patch‐clamp study, overexpression of Bcl‐2 in mitochondria of cultured cells suppressed calcium activation of a high conductance channel that may underlie the permeability transition. All other single channel parameters were identical when multiple conductance channel activities of mitochondria from control and Bcl‐2 overexpressing cells were compared. Bcl‐2 forms channels in artificial membranes; however, no novel channel activities could be linked to Bcl‐2 overexpression, suggesting Bcl‐2 does not form channels in native inner membranes of mitochondria.
A new phosphoglycerolipid, ‘phosphatidylglucose’, found in human cord red cells by multi‐reactive monoclonal anti‐i cold agglutinin, mAb GL‐1/GL‐2
Authors: Nagatsuka Yasuko , Kasama Takeshi , Ohashi Yoko , Uzawa Jun , Ono Yasushi , Shimizu Kazufumi , Hirabayashi Yoshio
Published: July 2002

Cord red cell membranes express many differentiation‐related molecules. To study such molecules, we have established human cell lines, termed GL‐1 and GL‐2, by the Epstein–Barr virus transformation method, both of which produce monoclonal anti‐i cold agglutinin [Y. Nagatsuka et al., Immunol. Lett. 46 (1995) 93–100]. Thin layer chromatography immunoblotting analysis revealed that these antibodies had broad specificities reacting with a variety of glycolipid antigens. Of the immunoreactive lipid antigens, a new phosphoglycerolipid containing glucose from human cord red cells was found. The isolated lipid was unstable to alkaline hydrolysis and contained glucose as a sole sugar. Secondary ion mass spectrum–collision‐induced dissociation mass spectrometric analysis of this lipid gave the main molecular ion peak at m/z 885 corresponding to phosphatidylhexose. This antigen was susceptible to phospholipases A2, C and D but resistant to phosphatidylinositol‐specific phospholipase C. Two‐dimensional nuclear magnetic resonance spectroscopy confirmed that glucose is linked to the sn‐glycerol 3‐phosphate residue with a β‐anomeric configuration. Based upon these combined results, we identified this lipid as phosphatidyl‐β‐D‐glucose. This is the first report showing the presence of the glucosylated glycerophospholipid in mammalian sources.
Up‐regulation of a constitutively active form of the β2‐adrenoceptor by sustained treatment with inverse agonists but not antagonists
Authors: MacEwan David J. , Milligan Graeme
Published: July 2002

In neuroblastoma × glioma hybrid, NG108‐15, cells transfected to stably express a constitutively active mutant (CAM) form of the human R2‐adrenoceptor, the β‐adrenoceptor ligands sotalol and betaxolol functioned as inverse agonists as they reduced basal adenylyl cyclase activity whereas the antagonists dihydroalprenolol and propranolol did not. Maintained presence of the CAM β
2‐adrenoceptor inverse agonists but not the antagonists in the culture medium of the cells resulted in a substantial, concentration‐dependent, up‐regulation of the CAMβ
2‐adrenoceptor. Up‐regulation of the CAMβ
2‐adrenoceptor by the inverse agonists was prevented by co‐incubation of the cells with either propranolol or dihydroalprenolol. Neither maintained elevation of cAMP levels nor the inhibition of adenylyl cyclase activity altered the ability of the inverse agonist ligands to cause receptor up‐regulation.
The adeno‐associated virus Rep78 major regulatory protein forms multimeric complexes and the domain for this activity is contained within the carboxy‐half of the molecule
Authors: Hermonat Paul L , Batchu Ramesh B
Published: July 2002

The adeno‐associated virus (AAV) encoded Rep78 is a multifunctional protein which is able to regulate transcription, is required for AAV DNA replication, and appears necessary for site specific integration of AAV DNA into human chromosome 19. Being analogous to the large T antigen, the replication protein of polyomaviruses which is known to homo‐multimerize, it seemed likely that the Rep78 protein would also interact with itself to carry out at least some of its functions. Furthermore, in electrophoretic mobility shift assay studies by many laboratories on Rep78/68 protein interaction with AAV terminal repeat DNA it has been noticed that multiple high bands often result. These data suggest Rep78‐Rep78 interaction. In this study it is directly demonstrated that Rep78 is able to form multimeric complexes as measured by West‐Western and chemical cross‐linking assays. Furthermore, using an amino‐truncated Rep78 protein, it is demonstrated that the Rep78 homo‐multimerization domain is contained within the carboxy‐half of the protein.
Nuclear translocation of PKCζ during ischemia and its inhibition by wortmannin, an inhibitor of phosphatidylinositol 3‐kinase
Authors: Mizukami Yoichi , Hirata Takao , Yoshida Ken-ichi
Published: July 2002

Protein kinase Cζ (PKCζ), a member of the atypical PKC subgroup, is insensitive to Ca2+, diacylglycerol, and phorbol esters, but is activated by phospholipids such as phosphatidylinositol‐3,4,5‐triphosphate, a product of phosphatidylinositol 3‐kinase (PI3‐kinase). Here we show that PKCζ translocates from the cytosol to the 1000×g pellet (nuclear‐myofibrillar) fraction during ischemia for 40 min in Langendorff‐perfused rat hearts. In addition, immunohistochemical observation shows that ischemia induces the translocation of PKCζ to the nucleus. The nuclear translocation during ischemia is inhibited in a dose‐dependent manner by wortmannin (10−9–10−7 M), an inhibitor of PI3‐kinase.
A human leptin mutant induces weight gain in normal mice
Authors: Verploegen Sandra A.B.W. , Plaetinck Geert , Devos René , Van der Heyden José , Guisez Yves
Published: July 2002

© 1997 Federation of European Biochemical Societies.
Insulin activates protein kinase B, inhibits glycogen synthase kinase‐3 and activates glycogen synthase by rapamycin‐insensitive pathways in skeletal muscle and adipose tissue
Authors: Cross Darren A.E. , Watt Peter W. , Shaw Morag , van der Kaay Jeroen , Downes C.Peter , Holder Julie C. , Cohen Philip
Published: July 2002

Insulin stimulated protein kinase Bα (PKBα) more than 10‐fold and decreased glycogen synthase kinase‐3 (GSK3) activity by 50±10% in skeletal muscle and adipocytes. Rapamycin did not prevent the activation of PKB, inhibition of GSK3 or stimulation of glycogen synthase up to 5 min. Thus rapamycin‐insensitive pathways mediate the acute effect of insulin on glycogen synthase in the major insulin‐responsive tissues. The small and very transient effects of EGF on phosphatidylinositol (3,4,5)P3 PKBα and GSK3 in adipocytes, compared to the strong and sustained effects of insulin, explains why EGF does not stimulate glucose uptake or glycogen synthesis in adipocytes
Rho effectors and reorganization of actin cytoskeleton
Authors: Narumiya Shuh , Ishizaki Toshimasa , Watanabe Naoki
Published: July 2002

The small GTPase Rho regulates several actomyosin‐based cellular processes such as cell adhesion, cytokinesis and contraction. The biochemical mechanisms of these actions remain unknown. Recently, several GTP‐Rho binding proteins were isolated. Among them, p140mDia and p160ROCK appear to work as Rho effectors mediating its action on the cytoskeleton. p140mDia induces actin polymerization by recruiting an actin binding protein, profilin, to the site of Rho action. p160ROCK induces focal adhesions and stress fibers by activating integrin and clustering them by the use of myosin‐based contractility.
A novel activity for a group of sesquiterpene lactones: inhibition of aromatase
Authors: Blanco Javier G. , Gil Roberto R. , Alvarez Cecila I. , Patrito Luis C. , Genti-Raimondi Susana , Flury Alfredo
Published: July 2002

A group of eleven sesquiterpene lactones isolated from different Asteraceae species from north‐western Argentina were investigated for their inhibitory action on the estrogen biosynthesis. Seven of them, of different skeleton types, were found to inhibit the aromatase enzyme activity in human placental microsomes, showing IC50 values ranging from 7 to 110 μM. The most active were the guaianolides 10‐epi‐8‐deoxycumambrin B (compound 1), dehydroleucodin (compound 2) and ludartin (compound 3). These compounds were competitive inhibitors with an apparent K
i=4 μM, K
i=21 μM and K
i=23 μM, respectively. Compounds 1 and 2 acted as type II ligands to the heme iron present in the active site of aromatase cytochrome P450 (P450arom). Besides, all of them failed to affect the cholesterol side‐chain cleavage enzyme activity on human placental mitochondrias. This is the first report on the aromatase inhibitory activity of this group of natural compounds.
Unique protein kinase C profile in mouse oocytes: lack of calcium‐dependent conventional isoforms suggested by rtPCR and Western blotting
Authors: Gangeswaran Rathi , Jones Keith T
Published: July 2002

rtPCR and Western blotting were used to determine which members of the PKC family are present in both immature and mature mouse oocytes. Using isoform‐specific PCR primers and antibodies PKC‐δ and ‐λ were detected while such techniques failed to observe the conventional isoforms of PKC‐α, ‐β, ‐γ. This isoform profile was confirmed using an alternative PCR strategy, which allowed discrimination of PCR products derived from conventional and novel PKC isoforms. In addition PKC‐ϵ, ‐η, ‐θ and ‐ζ were not detected by rtPCR. These results suggest that the predominant isoforms in oocytes are PKC‐δ and ‐λ.
Glucose exerts opposite effects on mRNA versus protein and activity levels of Pde1, the low‐affinity cAMP phosphodiesterase from budding yeast, Saccharomyces cerevisiae
Authors: Wera Stefaan , Ma Pingsheng , Thevelein Johan M
Published: July 2002

In budding yeast (Saccharomyces cerevisiae), a low‐affinity phosphodiesterase, Pde1, and a high‐affinity phosphodiesterase, Pde2, are responsible for the degradation of cAMP. Addition of glucose to glycerol‐grown yeast cells is known to cause a transient increase in the cAMP level and recent work has indicated a specific involvement of Pde1 in this response. In this work we show that glucose addition induces the accumulation to high levels of mRNA encoding Pde1. This increase continues for at least 8 hours and is due to enhanced transcription of the PDE1 gene, since glucose addition does not change the stability of the Pde1 mRNA. Surprisingly, using an assay method specific for Pde1, we observed that the activity of Pde1 remains constant and finally decreases several‐fold during the same period. In addition, this activity profile closely follows the Pde1 protein level as judged from Western blotting with antibodies directed against Pde1. Experiments using cycloheximide, a general inhibitor of translation, allow to exclude the possibility of a futile cycle of Pde1 synthesis and degradation. Hence, glucose addition appears to trigger an increase in PDE1 gene transcription together with a specific inhibition of the translation of Pde1 mRNA.
The endogenous cannabinoid anandamide is a lipid messenger activating cell growth via a cannabinoid receptor‐independent pathway in hematopoietic cell lines
Authors: Derocq J.-M , Bouaboula M , Marchand J , Rinaldi-Carmona M , Ségui M , Casellas P
Published: July 2002

The effect of anandamide, an endogenous ligand for central (CB1) and peripheral (CB2) cannabinoid receptors, was investigated on the growth of the murine IL‐6‐dependent lymphoid cell line B9 and the murine IL‐3‐dependent myeloblastic cell line FDC‐P1. In conditions of low serum level, anandamide potentiated the growth of both cytokine‐dependent cell lines. Comparison with other fatty acid cannabinoid ligands such as (R)‐methanandamide, a ligand with improved selectivity for the CB1 receptor, or palmitylethanolamide, an endogenous ligand for the CB2 receptor, showed a very similar effect, suggesting that cell growth enhancement by anandamide or its analogs could be mediated through either receptor subtype. However, several lines of evidence indicated that this growth‐promoting effect was cannabinoid receptor‐independent. First, the potent synthetic cannabinoid agonist CP 55940, which displays high affinity for both receptors, was inactive in this model. Second, SR 141716A and SR 144528, which are potent and specific antagonists of CB1 and CB2 receptors respectively, were unable, alone or in combination, to block the anandamide‐induced effect. Third, inactivation of both receptors by pretreatment of cells with pertussis toxin did not affect the potentiation of cell growth by anandamide. These data demonstrated that neither CB1 nor CB2 receptors were involved in the anandamide‐induced effect. Moreover, using CB2‐transfected Chinese hamster ovary cells, we demonstrated that after complete blockade of the receptors by the specific antagonist SR 144528, anandamide was still able to strongly stimulate a mitogen‐activated protein (MAP) kinase activity, clearly indicating that the endogenous cannabinoid can transduce a mitogenic signal in the absence of available receptors. Finally, arachidonic acid, a structurally related compound and an important lipid messenger without known affinity for cannabinoid receptors, was shown to trigger MAP kinase activity and cell growth enhancement similar to those observed with anandamide. These findings provide clear evidence for a functional role of anandamide in activating a signal transduction pathway leading to cell activation and proliferation via a non‐cannabinoid receptor‐mediated process.
IncI1 plasmid R64 encodes the ArsR protein that alleviates type I restriction
Authors: Rastorguev Sergei M , Zavilgelsky Gennadii B , Tchurikov Nickolai A
Published: July 2002

The host‐controlled EcoK restriction of unmodified phage λ was five‐fold alleviated in the wild‐type Escherichia coli strain K12 carrying the R64 plasmid of the incompatibility group I1. The relevant gene was mapped between the origin of vegetative replication (rep, oriV) and the tet
r gene about 60 kbp downstream from the origin of transfer, oriT. We cloned this gene inside the 613 bp long EcoRI‐PstI fragment and sequenced it. Only one 351 bp long open reading frame (ORF) starting at 124 bp from the beginning of the insert was found in the sequence. Computer search in the current databases revealed that the putative protein is identical to the ArsR protein specified by the IncFI plasmid R773. ArsR is a repressor of the arsenical resistance (ars) operon, arsRDABC. There are no arsABC genes in the R64 plasmid since plasmid R64‐ (or pSR8)‐mediated resistance of E. coli K12 cells to the arsenicals arsenate and arsenite was not detected. The gene arsR and the antirestriction genes ard (ardA and ardB) are non‐homologous. However, comparison of the deduced amino acid sequence of ArsR with the ArdA and ArdB sequences revealed only one small region of similarity, a 9 amino acid motif found in different antirestriction proteins that is hypothesized to be an interaction site for antirestriction proteins with restriction endonucleases.
Peroxisome proliferator‐activated receptor γ1 (PPAR‐γ1) as a major PPAR in a tissue in which estrogen induces peroxisome proliferation
Authors: Ma Hongwen , Tam Quach Thanh , Kolattukudy Pappachan E.
Published: July 2002

Estradiol administration induces peroxisome proliferation and the production of 3‐hydroxy fatty acid pheromones in the uropygial glands of the duck, but not in the goose gland, which does not produce such pheromones. We isolated a peroxisome proliferator‐activated receptor (PPAR)γ1 cDNA from a duck uropygial gland cDNA library. Northern blots revealed two transcripts, PPARγ1 and γ2, and showed that PPARγ was expressed at higher levels than PPARα in the uropygial gland of the duck. Although PPARγ2 was expressed in both duck and goose uropygial gland, PPARγ1 was expressed only in the duck gland, which responds to estrogen by peroxisome proliferation. In NIH 3T3 transfected cells, PPARγ1 was activated by peroxisome proliferators such as Wy‐14 643, clofibric acid and Ly‐171 883 causing induction of the target marker gene. By cotransfection with a plasmid containing α‐cis‐retinoic acid receptor RXRα, the induction increased up to 9‐fold. These results suggest that PPARγ1 may be involved in peroxisome proliferation while PPARγ2 may be involved in lipid metabolism.
Evidence for the specific interaction of a lipid molecule with rhodopsin which is altered in the transition to the active state metarhodopsin II1
Authors: Beck Mareike , Siebert Friedrich , Sakmar Thomas P
Published: July 2002

Comparing the FTIR difference spectra of the rhodopsin→metarhodopsin II transition in membranes and in dodecylmaltoside detergent, characteristic variations are observed between 1715 and 1750 cm−1. By repeating the measurements with the rhodopsin mutant D83N/E122Q, the spectral variation between the samples in membranes versus detergent could be assigned to a difference band at 1743(+)/1724(−) cm−1, which does not exhibit a deuteration‐induced downshift. We provide evidence that this band is probably caused by the C=O stretch of only one ester group of one lipid molecule. This group interacts with the dark state of rhodopsin, whereas in metarhodopsin II, the lipid molecule behaves as if it were in the bulk lipid phase.
Replication in the phloem is not necessary for efficient vascular transport of tobacco mosaic tobamovirus
Authors: Susi Petri
Published: July 2002

Plant viruses move systemically from one leaf to another via phloem. However, the viral functions needed for systemic movement are not fully elucidated. An experimental system was designed to study the effects of low temperature on the vascular transport of the tobacco mosaic tobamovirus (TMV). Vascular transport of TMV from lower inoculated leaves to upper non‐inoculated leaves via a stem segment kept at low temperature (4°C) was not affected. On the other hand, several experiments were performed on tobacco leaves to demonstrate that virus replication did not occur at the same temperature. The data suggest that replication of TMV in the phloem of wild‐type tobacco plants is not necessary for the vascular transport of TMV, and that the virus moves with photoassimilates as suggested previously.
Efficient amplification and direct sequencing of mouse variable regions from any immunoglobulin gene family
Authors: Chardès Thierry , Villard Sylvie , Ferrières Gaelle , Piechaczyk Martine , Cerutti Martine , Devauchelle Gérard , Pau Bernard
Published: July 2002

We have designed two original sets of oligonucleotide primers hybridizing the relatively conserved motifs within the immunoglobulin signal sequences of each of the 15 heavy chain and 18 kappa light chain gene families. Comparison of these 5′ primers with the immunoglobulin signal sequences referenced in the Kabat database suggests that these oligonucleotide primers should hybridize with 89.4% of the 428 mouse heavy chain signal sequences and with 91.8% of the 320 kappa light chain signal sequences with no mismatch. Following PCR amplification using the designed primers and direct sequencing of the amplified products, we obtained full‐length variable sequences belonging to major (VH1, VH2, VH3, Vκ1 and Vκ21) but also small‐sized (VH9, VH14, Vκ2, Vκ9A/9B, Vκ12/13, Vκ23 and Vκ33/34) gene families, from nine murine monoclonal antibodies. This strategy could be a powerful tool for antibody sequence assessment whatever the V gene family before humanization of mouse monoclonal antibody or identification of paratope‐derived peptides.
Binding of calcium in the EF‐hand of Escherichia coli lytic transglycosylase Slt35 is important for stability
Authors: van Asselt Erik J , Dijkstra Bauke W
Published: July 2002

The Escherichia coli lytic transglycosylase Slt35 contains a single metal ion‐binding site that resembles EF‐hand calcium‐binding sites. The Slt35 EF‐hand is only the second observation of such a domain in a prokaryotic protein. Two crystal structures at 2.1 Å resolution show that both Ca2+ ions and Na+ ions can bind to the EF‐hand domain, but in subtly different configurations. Heat‐induced unfolding studies demonstrate that Ca2+ ions are preferentially bound, and that only Ca2+ ions significantly increase the melting temperature of Slt35. This shows that the EF‐hand calcium‐binding domain is important for the stability of Slt35.
The hydrophobic lock‐and‐key intersubunit motif of glutathione transferase A1‐1: implications for catalysis, ligandin function and stability
Authors: Sayed Yasien , Wallace Louise A. , Dirr Heini W.
Published: July 2002

A hydrophobic lock‐and‐key intersubunit motif involving a phenylalanine is a major structural feature conserved at the dimer interface of classes alpha, mu and pi glutathione transferases. In order to determine the contribution of this subunit interaction towards the function and stability of human class alpha GSTA1‐1, the interaction was truncated by replacing the phenylalanine ‘key’ Phe‐51 with serine. The F51S mutant protein is dimeric with a native‐like core structure indicating that Phe‐51 is not essential for dimerization. The mutation impacts on catalytic and ligandin function suggesting that tertiary structural changes have occurred at/near the active and non‐substrate ligand‐binding sites. The active site appears to be disrupted mainly at the glutathione‐binding region that is adjacent to the lock‐and‐key intersubunit motif. The F51S mutant displays enhanced exposure of hydrophobic surface and ligandin function. The lock‐and‐key motif stabilizes the quaternary structure of hGSTA1‐1 at the dimer interface and the protein concentration dependence of stability indicates that the dissociation and unfolding processes of the mutant protein remain closely coupled.
Infertility and pregnancy outcome in female genital tuberculosis
Authors: Tripathy Sindhu N. , Tripathy Satchida N.
Published: July 2002

Objectives: The aim of the study is to determine the prevalence rate, identify risk factors not conducive to pregnancy, and outcomes with regard to follow‐up treatment. Methods: This prospective study was carried out from 1988 to 2001, in the Department of Obstetrics and Gynecology in the S.C.B. Medical College, Cuttack, and in the private clinics of both the authors in Cuttack, India. The incidence of tuberculosis in infertility and tubal factor infertility cases were determined. The incidence of infertility in genital tuberculosis (GT) was examined and a total of 97 cases of infertility with genital tuberculosis were studied. The patients were given short course chemotherapy and prednisolone (wherever feasible). The risk factors not conducive to pregnancy were identified. Results: The incidence of GT in infertility and tubal factor infertility were 3 and 41%, respectively. The incidence of infertility in GT was 58%. All patient were symptom free and no evidence of tuberculosis was detected after the end of the chemotherapy. The risk factors not conducive to pregnancy were secondary amenorrhea, no endometrium on curettage, and negative chromopertubation. The conception rate is low, i.e. 19.2%, the live birth rate being still low, i.e. 7.2%. Conclusion: The outcome of infertility in GT is not very optimistic. IVF and ET offers some hope to these unfortunate women.
Spatial and temporal gene expression in chondrogenesis during fracture healing and the effects of basic fibroblast growth factor
Authors: Nakajima Fumitake , Ogasawara Akira , Goto Ken‐Ichiro , Moriya Hideshige , Ninomiya Yoshifumi , Einhorn Thomas A. , Yamazaki Masashi
Published: July 2002

Chondrogenesis is an essential component of endochondral fracture healing, though the molecular and cellular events by which it is regulated have not been fully elucidated. In this study, we used a rat model of closed fracture healing to determine the spatial and temporal expression of genes for cartilage‐specific collagens. Furthermore, to determine the effects of basic fibroblast growth factor (bFGF) on chondrogenesis in fracture healing, we injected 100 μg recombinant human bFGF into the fracture site immediately after fracture.

In normal calluses, pro‐α(II) collagen mRNA (COL2A1) was detected in proliferative chondrocytes beginning on day 4 after the fracture, and pro‐α(X) collagen mRNA (COL10A1) in hypertrophic chondrocytes beginning on day 7. In FGF‐injected calluses, the cartilage enlarged in size significantly. On day 14, both COL2A1‐and COL10A1‐expressing cells were more widely distributed, and the amounts of COL2A1 and COL10A1 mRNAs were both approximately 2‐fold increased when compared with uninjected fractures. Temporal patterns of expression for these genes were, however, identical to those found in normal calluses. The number of proliferating cell nuclear antigen‐positive cells was increased in the non‐cartilaginous area in the bFGF‐injected calluses by day 4.

The present molecular analyses demonstrate that a single injection of bFGF enhances the proliferation of chondroprogenitor cells in fracture callus, and thus contributes to the formation of a larger cartilage. However, maturation of chondrocytes and replacement of the cartilage by osseous tissue are not enhanced by exogenous bFGF, and this results in the prolonged cartilaginous callus phase. We conclude that, in the healing of closed fractures of long bones, exogenous bFGF has a capacity to enlarge the cartilaginous calluses, but not to induce more rapid healing. © 2001 Orthopaedic Research Society. Punlished by Elsevier Science Ltd. All rights reserved.
Brain‐derived neurotrophic factor stimulates neurite outgrowth in a calretinin‐enriched neuronal culture system
Authors: Iwasaki Katsunori , Isaacs Krystyna r , Jacobowitz David M.
Published: July 2002

A calretinin enriched cell culture system which comprised approximately 40% of the total neuronal population of the E14 rat embryo was established from the region of the thalamic eminence (TE), and the effects of several neurotrophins on the neurite growth of calretinin‐immunoreactive (CR‐IR) neurons was investigated. A 4‐day treatment of BDNF significantly increased the ratio of CR‐IR to microtubule‐associated protein 2‐immunoreactive neurons at concentrations between 50 and 250 ng\ml. IGF‐I at 100 ng\ml and TGF‐ at 250 ng\ml also increased this ratio. None of the neurotrophins examined increased the number of primary neurites. BDNF did, however, increase the number of secondary neurites. BDNF‐treated primary and secondary neurites were also significantly longer than neurites from neurons in control cultures. IGF‐I elicited an increase in primary neurite length, but did not affect either number or length of secondary neurites. TGF‐α had no effect on either number or length of the primary and secondary neurites.

These results indicate that the maturation and development of CR‐IR neurites is specifically affected by BDNF. It is suggested that BDNF increases the CR concentration above the threshold of detection by immunohistochemistry in cells and stimulates the sprouting of secondary CR‐IR neurites.
Expression of glial cell line‐derived neurotrophic factor in the brain and cerebrospinal fluid of the developing rat
Authors: Ikeda Tomoaki , Xia Xiao Y. , Xia Yi X. , Ikenoue Tsuyomu , Choi Ben H.
Published: July 2002

Expression of GDNF in developing rat brain from PND 1 to 14 and on PND 21 was examined immunocytochemically. At PND 1, intense diffuse immunoreactivity was noted within the cytoplasm of a diverse group of neuronal and nonneuronal cells, including choroid plexus epithelial cells, ependymal cells, tanycytes of the third ventricle, and cellular elements in the subarachnoid compartment. GDNF expression became more localized among these cells from PND 7–14 and was almost undetectable by PND 21. Although GDNF‐positive small glial cells were scattered within the cerebral cortical plate and the striatum already at PND 1, GDNF expression among astroglial cells within the corpus callosum and in the white matter adjacent to the lateral ventricles was more prominent between PND 5 and 8. GDNF expression among the pyramidal neurons of the cerebral cortex was evident relatively early in the postnatal period, but the neurons of the hippocampus and thalamus showed more intense immunoreactivity at later periods between PND 8–14. ELISA of the CSF revealed a rapid rise in GDNF levels from 71.4±10.9 pg/ml (mean±S.E.M.) at PND 1 to peak levels of 138.4±18.5, 135.1±5.4 and 132.9±8.0 pg/ml, at PND 5, 7 and 9, respectively. Peak CSF levels of GDNF occurred when GDNF expression was intense within astroglial cells in the corpus callosum and cerebral white matter. Thereafter, the levels gradually decreased to 76.5±9.7 pg/ml at PND 21. The widespread expression of GDNF among different cellular elements in the developing brain suggest that GDNF probably plays diverse functional roles in many different neuronal systems in addition to its known effects on the dopaminergic system. Developmental shifts in GDNF expression further suggest that GDNF may be of critical importance at different stages of brain growth and differentiation.
Long-term effect on outcome of clozapine in chronic therapy-resistant schizophrenic patients
Authors: Lindström LH , Lundberg T
Published: July 2002

Controlled clinical trials have shown that the atypical neuroleptic clozapine is highly effective in schizophrenic patients who are unresponsive to conventional neuroleptic agents. The long-term outcome of clozapine treatment was studied in 122 patients who were treated between 1974 and 1991. The mean duration of treatment was 5.2 years. At follow-up, 74 patients (61%) were still receiving clozapine. Only 11 patients discontinued treatment because of adverse events and eight because of poor compliance. Clinical improvement was seen in 87% of patients; 40% had resumed employment after 2 years' treatment. About one-third of patients who received clozapine for 5–10 years continued to improve during this time; this was probably because of a continuing process of socialisation. Thus, clozapine offers important advantages in schizophrenic patients resistant to conventional neuroleptics in terms of long-term efficacy and lack of extrapyramidal side-effects.
Soluble IL-6 receptors in the serum and cerebrospinal fluid of paranoid schizophrenic patients
Authors: Müller N , Dobmeier P , Empl M , Riedel M , Schwarz M , Ackenheil M
Published: July 2002

Soluble Interleukin-6 receptor (sIL-6R) levels are strongly related to the levels of Interleukin-6 (IL-6), and sIL-6Rs increase the immune activating properties of IL-6. We estimated sIL-6R serum levels in 25 schizophrenic patients and 25 healthy controls. In the patients, SIL-6R-CSF levels were also measured. The psychopathology was rated according to the AMDP system. We found a significant correlation between serum and cerebrospinal fluid (CSF) levels of sIL-6R, suggesting that serum levels may be a meaningful marker for the central action of sIL-6R. Moreover, significant correlations between the paranoid-hallucinatory syndrome and sIL-6R levels both in serum and CSF were observed. This finding suggests that IL-6 plays a role in the paranoid-hallucinatory symptomatology in schizophrenia. This can be understood regarding the influence of IL-6 to the catecholaminergic neurotransmission. The downregulating effects of neuroleptic treatment to sIL-6R demonstrate that the sIL-6R levels are decreased in the whole group of schizophrenic patients compared to controls.
Hidden dimensions: the stability and structure of regional political cleavages in Poland
Authors: Zarycki Tomasz , Nowak Andrzej
Published: July 2002

The main aim of this paper is to show that behind the complex and fluid surface dimension of the Polish political scene it is possible to reveal a relatively stable two-dimensional structure of deep political identities. The study is based on the analysis of ecological data — the results of elections from 1990 to 1997. The results prove the exceptional stability of the Polish electoral geography and appear to be consistent with other studies based mainly on the survey data. The use of the spatial data allowed us to reveal the historical and spatial determinants of electoral behavior in Poland.
The middle class in the Czech reforms: the interplay between policies and social stratification
Authors: Večerník Jiřı´
Published: July 2002

Following the assumption that the middle class is important in the transition from communism to a democratic market society as its leading actor and guarantor of social integration, this paper presents evidence of the benefits it has gained from economic and social reforms. As these have turned out to be rather modest, it is likely that the middle class will be more reserved in its political support for a rigorous transformation. Special attention is given to the distribution and redistribution of income in which the middle seems to have been neglected, in comparison to the lowest and upper strata which have become better off. Evidence based on official statistics is complemented by a survey of attitudes which document both the still strong endorsement of the middle class with regard to the implemented changes and a certain split between the ‘old’ and ‘new' component of this important social category.
Nitric oxide: not just a negative inotrope
Authors: Sarkar David , Vallance Patrick , Harding Sian E.
Published: July 2002

Nitric oxide (NO) appears to play a role in modulating cardiac function in both health and disease. Early studies in isolated rodent cardiac myocytes demonstrated a depressant effect of NO supplied by NO donors (exogenous) as well as NO generated within myocytes (endogenous). There is increasing evidence for a functional NO generating system within the human myocardium, which appears upregulated in certain disease states. Induction of the high output nitric oxide synthase isoform (iNOS) has been demonstrated in the failing myocardium, though its functional significance remains unproven. More recently published data have contradicted the notion that NO acts solely as a negative inotrope demonstrating positive inotropy in both isolated rodent and human ventricular myocytes in response to a range of NO donors. Different NO donors have different NO release kinetics and generate a range of NO species (NO·, NO+ and NO−) which may interact at a number of subcellular targets. The observed response of any cardiac preparation to an NO donor represents the net effect of activation of different effector targets and may explain the contradictory reported effects of NO. To realise the therapeutic potential of NO will require specific targeting at a subcellular level.
A simple, yet novel hybrid‐dynamic conformal arc therapy planning via flattening filter‐free beam for lung stereotactic body radiotherapy
Authors: Pokhrel Damodar , Halfman Matthew , Sanford Lana
Published: April 2020

Purpose

Due to multiple beamlets in the delivery of highly modulated volumetric arc therapy (VMAT) plans, dose delivery uncertainties associated with small‐field dosimetry and interplay effects can be concerns in the treatment of mobile lung lesions using a single‐dose of stereotactic body radiotherapy (SBRT). Herein, we describe and compare a simple, yet clinically useful, hybrid 3D‐dynamic conformal arc (h‐DCA) planning technique using flattening filter‐free (FFF) beams to minimize these effects.

Materials and Methods

Fifteen consecutive solitary early‐stage I‐II non‐small‐cell lung cancer (NSCLC) patients who underwent a single‐dose of 30 Gy using 3–6 non‐coplanar VMAT arcs with 6X‐FFF beams in our clinic. These patients’ plans were re‐planned using a non‐coplanar hybrid technique with 2–3 differentially‐weighted partial dynamic conformal arcs (DCA) plus 4–6 static beams. About 60–70% of the total beam weight was given to the DCA and the rest was distributed among the static beams to maximize the tumor coverage and spare the organs‐at‐risk (OAR). The clinical VMAT and h‐DCA plans were compared via RTOG‐0915 protocol for conformity and dose to OAR. Additionally, delivery efficiency, accuracy, and overall h‐DCA planning time were recorded.

Results

All plans met RTOG‐0915 requirements. Comparison with clinical VMAT plans h‐DAC gave better target coverage with a higher dose to the tumor and exhibited statistically insignificance differences in gradient index, D2cm, gradient distance and OAR doses with the exception of maximal dose to skin (P = 0.015). For h‐DCA plans, higher values of tumor heterogeneity and tumor maximum, minimum and mean doses were observed and were 10%, 2.8, 1.0, and 2.0 Gy, on average, respectively, compared to the clinical VMAT plans. Average beam on time was reduced by a factor of 1.51. Overall treatment planning time for h‐DCA was about an hour.

Conclusion

Due to no beam modulation through the target, h‐DCA plans avoid small‐field dosimetry and MLC interplay effects and resulting in enhanced target coverage by improving tumor dose (characteristic of FFF‐beam). The h‐DCA simplifies treatment planning and beam on time significantly compared to clinical VMAT plans. Additionally, h‐DCA allows for the real time target verification and eliminates patient‐specific VMAT quality assurance; potentially offering cost‐effective, same or next day SBRT treatments. Moreover, this technique can be easily adopted to other disease sites and small clinics with less extensive physics or machine support.
Idebenone increases chance of stabilization/recovery of visual acuity in OPA1‐dominant optic atrophy
Authors: Romagnoli Martina , La Morgia Chiara , Carbonelli Michele , Di Vito Lidia , Amore Giulia , Zenesini Corrado , Cascavilla Maria Lucia , Barboni Piero , Carelli Valerio
Published: April 2020

We previously documented that idebenone treatment in OPA1‐Dominant Optic Atrophy (OPA1‐DOA) led to some degrees of visual improvement in seven patients. We here present the results of a cohort study, which investigated the effect of off‐label idebenone administration in a larger OPA1‐DOA group compared with untreated patients. Inclusion criteria were: OPA1‐DOA clinical and molecular diagnosis, baseline visual acuity (VA) greater than/equal to counting fingers and treatment duration greater than 7 months. We found a significant difference between the last visit and baseline VA in favor of stabilization/recovery in idebenone‐treated as compared to untreated patients. This effect was retained after controlling for confounders.