julilili42/papers huggingface dataset:
α1D (Cav1.3) Subunits Can Form L-type Ca2+ Channels Activating at Negative Voltages
2001-06-01

In cochlea inner hair cells (IHCs), L-type Ca(2+) channels (LTCCs) formed by alpha1D subunits (D-LTCCs) possess biophysical and pharmacological properties distinct from those of alpha1C containing C-LTCCs. We investigated to which extent these differences are determined by alpha1D itself by analyzing the biophysical and pharmacological properties of cloned human alpha1D splice variants in tsA-201 cells. Variant alpha1D(8A,) containing exon 8A sequence in repeat I, yielded alpha1D protein and L-type currents, whereas no intact protein and currents were observed after expression with exon 8B. In whole cell patch-clamp recordings (charge carrier 15-20 mm Ba(2+)), alpha1D(8A) - mediated currents activated at more negative voltages (activation threshold, -45.7 versus -31.5 mV, p < 0.05) and more rapidly (tau(act) for maximal inward currents 0.8 versus 2.3 ms; p < 0.05) than currents mediated by rabbit alpha1C. Inactivation during depolarizing pulses was slower than for alpha1C (current inactivation after 5-s depolarizations by 90 versus 99%, p < 0.05) but faster than for LTCCs in IHCs. The sensitivity for the dihydropyridine (DHP) L-type channel blocker isradipine was 8.5-fold lower than for alpha1C. Radioligand binding experiments revealed that this was not due to a lower affinity for the DHP binding pocket, suggesting that differences in the voltage-dependence of DHP block account for decreased sensitivity of D-LTCCs. Our experiments show that alpha1D(8A) subunits can form slowly inactivating LTCCs activating at more negative voltages than alpha1C. These properties should allow D-LTCCs to control physiological processes, such as diastolic depolarization in sinoatrial node cells, neurotransmitter release in IHCs and neuronal excitability.
The C3435T mutation in the human MDR1 gene is associated with altered efflux of the P-glycoprotein substrate rhodamine 123 from CD56+ natural killer cells
2001-06-01

P-glycoprotein (PGP) is a membrane protein which determines drug disposition in humans (e.g. digoxin). It is also expressed in various leukocyte lineages with highest expression in CD56+ natural killer cells. Recently, a polymorphism in exon 26 (C3435T) of this gene was shown to correlate with intestinal PGP expression and function in humans. Carriers homozygous for this polymorphism (TT) showed more than two-fold lower PGP expression and higher digoxin plasma concentrations compared to the CC group. However, it is not known whether this mutation in the MDR1 gene is also associated with altered PGP function in peripheral blood cells. We therefore assessed efflux of the PGP-substrate rhodamine 123 from CD56+ natural killer cells. Leukocytes were isolated from whole blood of 10 CC, 10 CT and 11 TT healthy Caucasian individuals. Using flow cytometry, rhodamine fluorescence was determined in CD56+ cells. Moreover, MDRI mRNA was quantified in leukocytes by real-time polymerase chain reaction. Subjects with CC genotype revealed a significantly lower rhodamine fluorescence (i.e. higher PGP function) compared to individuals with TT genotype (51.1 +/- 11.4% versus 67.5 +/- 9.5%, p < 0.01). Heterozygous individuals had an intermediate rhodamine fluorescence (61.4 +/- 6.3%). MDR1 mRNA normalized for cyclophilin was lowest in the TT population (1.29 +/- 1.01), intermediate in heterozygous subjects (1.60 +/- 0.76) and highest in the CC group (1.91 +/- 0.94; not significant). In summary, subjects being homozygous for C in position 3435 of the MDR1 gene have a more pronounced efflux of rhodamine from CD56+ natural killer cells and a higher MDR1 mRNA expression in leukocytes than subjects with the TT genotype. Measurement of rhodamine efflux using flow-cytometry from peripheral blood cells allows assessment of genetically determined differences in P-glycoprotein function.
Anaerobic Conditions Induce Expression of Polysaccharide Intercellular Adhesin in Staphylococcus aureus and Staphylococcus epidermidis
2001-06-01

ABSTRACT Products of the intercellular adhesion ( ica ) operon in Staphylococcus aureus and Staphylococcus epidermidis synthesize a linear β-1,6-linked glucosaminylglycan. This extracellular polysaccharide mediates bacterial cell-cell adhesion and is required for biofilm formation, which is thought to increase the virulence of both pathogens in association with prosthetic biomedical implants. The environmental signal(s) that triggers ica gene product and polysaccharide expression is unknown. Here we demonstrate that anaerobic in vitro growth conditions lead to increased polysaccharide expression in both S. aureus and S. epidermidis , although the regulation is less stringent in S. epidermidis . Anaerobiosis also dramatically stimulates ica -specific mRNA expression in ica - and polysaccharide - positive strains of both S. aureus and S. epidermidis. These data suggest a mechanism whereby ica gene expression and polysaccharide production may act as a virulence factor in an anaerobic environment in vivo.
Impact of the regulatory loci agr, sarA and sae of Staphylococcus aureus on the induction of α‐toxin during device‐related infection resolved by direct quantitative transcript analysis
2001-06-01

The cytotoxic α‐toxin (encoded by hla ) of Staphylococcus aureus is regulated by three loci, agr , sar A and sae , in vitro . Here, we assess the regulation of hla in a guinea pig model of device‐related infection by quantifying RNAIII (the effector molecule of agr ) and hla directly in exudates accumulating in infected devices without subculturing of the bacteria. LightCycler reverse transcription–polymerase chain reaction (RT–PCR) was used to quantify the transcripts. Strains RN6390 and Newman expressed considerably smaller amounts of RNAIII in the guinea pig than during in vitro growth. The residual RNAIII expression decreased during the course of infection and was negatively correlated with bacterial densities. As with RNAIII, the highest hla expression was detected in both strains early in infection. Even in strain Newman, a weak hla producer in vitro , a pronounced expression of hla was observed during infection. Likewise, four S. aureus isolates from cystic fibrosis (CF) patients expressed hla despite an inactive agr during device‐related infection as in the CF lung. Mutation of agr and sar A in strain Newman and RN6390 had no consequence for hla expression in vivo . In contrast, the mutation in sae resulted in severe downregulation of hla in vitro as well as in vivo . In conclusion, S. aureus seems to be provided with regulatory circuits different from those characterized in vitro to ensure α‐toxin synthesis during infections.
Allogeneic Cancellous Bone Graft and a Burch-Schneider Ring for Acetabular Reconstruction in Revision Hip Arthroplasty
2001-06-01

Acetabular reconstruction with use of morselized cryopreserved allogeneic cancellous bone graft and the Burch-Schneider ring can be highly successful in managing massive acetabular deficiencies in revision hip arthroplasty.
Efficacy and tolerability of a standardized willow bark extract in patients with osteoarthritis: randomized placebo‐controlled, double blind clinical trial
2001-06-01

Abstract This study assessed the clinical efficacy of a chemically standardized willow bark extract in the treatment of osteoarthritis. Willow bark extract, in a dose corresponding to 240 mg salicin/day, was compared with placebo in a 2‐week, double‐blind, randomized controlled trial. The primary outcome measure was the pain dimension of the WOMAC Osteoarthritis Index. Secondary outcome measures included the stiffness and physical function dimensions of the WOMAC, daily visual analogue scales (VAS) on pain and physical function, and final overall assessments by both patients and investigators. A total of 78 patients (39 willow bark extract, 39 placebo) participated in the trial. A statistically significant difference between the active treatment and the placebo group was observed in the WOMAC pain dimension ( d = 6.5 mm, 95% C.I. = 0.2–12.7 mm, p = 0.047); the WOMAC pain score was reduced by 14% from the baseline level after 2 weeks of active treatment, compared with an increase of 2% in the placebo group. The patient diary VAS confirmed this result, and likewise the final overall assessments showed superiority of the willow bark extract over the placebo (patients' assessment, p = 0.0002; investigators' assessment, p = 0.0073). It is concluded that the willow bark extract showed a moderate analgesic effect in osteoarthritis and appeared to be well tolerated. Copyright © 2001 John Wiley &amp; Sons, Ltd.
Molecular determinants of glioma cell migration and invasion
2001-06-01

Object. Migration and invasion are important prerequisites for the infiltrative and destructive growth patterns of malignant gliomas. Infiltrative growth prevents complete tumor resection and causes significant neurological morbidity and mortality. Methods. The authors assessed the expression of matrix metalloproteinases (MMPs) at messenger RNA and protein levels, MMP-2 and MMP-9 activities, and expression levels of a panel of anti- and proapoptotic proteins of the BCL-2 family. They then correlated their findings with α V β 3 integrin expression and the migratory and invasive potentials in 12 human malignant glioma cell lines. Multiple MMPs were expressed by most cell lines. The levels of MMP-2 and MMP-3 and the activities of MMP-2 and MMP-9 correlated with tumor cell invasion. Migration and invasion were also correlated. Although the expression levels of α V β 3 integrin did not predict migration or invasion, a neutralizing α V β 3 integrin antibody inhibited migration and invasion selectively in cell lines that contained a high level of α V β 3 integrin expression, thus indicating the important role of α V β 3 integrin for migration and invasion in this subset of cell lines. An expression pattern of BCL-2 family proteins that favor resistance to apoptosis was associated with enhanced migration, invasion, and MMP activity. Wild-type p53 cell lines migrated farther than mutant p53 cell lines. Conclusions. Activities of MMP-2 and MMP-9 are the best predictors of glioma cell invasion. The α V β 3 integrin mediates migration and invasion in a subset of glioma cell lines, but these processes do not depend on α V β 3 integrin expression. Antiapoptotic BCL-2 family protein expression is a predictor of efficient migration and invasion.
Injections of the NMDA Receptor Antagonist Aminophosphonopentanoic Acid into the Lateral Nucleus of the Amygdala Block the Expression of Fear-Potentiated Startle and Freezing
2001-06-01

NMDA receptors within the amygdala play an important role in the acquisition and expression of conditioned fear. Because amygdaloid injections of NMDA receptor antagonists did not block the expression of every behavioral sign of fear, a discussion arose as to whether amygdaloid NMDA receptors play different roles in different kinds of fear-conditioning tasks. To clarify the exact role of amygdaloid NMDA receptors, the present study measured the effects of amygdaloid NMDA receptor blockade on the two major animal models of conditioned fear. An experimental design was used that allowed simultaneous measurement of fear-potentiated startle and freezing during the same test session after animals had undergone identical training procedures. The present study clearly demonstrates that injections of the NMDA receptor antagonist AP-5 into the lateral nucleus of the amygdala significantly attenuated both behavioral fear responses (i.e., the amygdaloid NMDA receptors are necessary for the expression of fear-potentiated startle and freezing). The present results together with others from the literature indicate that NMDA receptors within the lateral amygdala are critically involved in normal synaptic transmission. It appears then that NMDA receptor antagonists may block the acquisition of fear conditioning by directly interfering with normal synaptic transmissions in the amygdala. Possible reasons for some discrepant results in earlier studies are also discussed.
Chemotaxis and transendothelial migration of CD34+hematopoietic progenitor cells induced by the inflammatory mediator leukotriene D4 are mediated by the 7-transmembrane receptor CysLT1
2001-06-01

Recent studies suggest that bone marrow (BM)–derived chemotactic mediators such as chemokines play key roles in hematopoietic stem cell trafficking. Lipid mediators, particularly leukotrienes, are involved in leukocyte chemotaxis during inflammation but have also been detected in the normal BM. Therefore, the effects of leukotrienes on hematopoietic progenitor cells were analyzed. Cysteinyl leukotrienes, particularly leukotriene D4 (LTD4), induced strong intracellular calcium fluxes and actin polymerization in mobilized and BM CD34+ progenitors. Chemotaxis and in vitro transendothelial migration of CD34+ and more primitive CD34+/CD38− cells were 2-fold increased by LTD4 at an optimum concentration of 25 to 50 nM. Accordingly, CD34+ cells expressed the 7-transmembrane LTD4 receptor CysLT1 by reverse transcriptase–polymerase chain reaction and Western blot. Effects of LTD4 were suppressed by the CysLT1 receptor antagonist MK-571 and reduced by pertussis toxin. In contrast, LTB4 induced strong responses only in mature granulocytes. LTD4-induced calcium fluxes were also observed in granulocytes but were not reduced by MK-571, suggesting that these effects were mediated by other receptors (eg, CysLT2) rather than by CysLT1. In addition, expression of 5-lipoxygenase, the key enzyme of leukotriene biosynthesis, was detected in both hematopoietic progenitor cells and mature leukocytes. The study concludes that the functionally active LTD4 receptor CysLT1 is preferentially expressed in immature hematopoietic progenitor cells. LTD4 released in the BM might regulate progenitor cell trafficking and could also act as an autocrine mediator of hematopoiesis. This would be a first physiologic effect of cysteinyl leukotrienes apart from the many known pathophysiologic actions related to allergy and inflammation.
Counting models of temporal discrimination
2001-06-01

A three-category task was employed to test counting models for temporal discrimination. Unlike former approaches, the present one is not based on Weber functions. Specifically, the proposed test does not require the implicit but, nevertheless, debatable assumption that the pulse rate of the internal clock is constant for different durations of the standard interval. Furthermore, the present approach does not necessitate specific distributional assumptions about the interpulse times. An experiment was conducted to evaluate the predictions of this generalized counting model. The results are consistent with predictions of the generalized counting model. A further analysis suggests that the pulse rate decreases as the duration of the standard interval increases.
Variation of heads of lateral pterygoid muscle and morphology of articular disc of human temporomandibular joint – anatomical and histological analysis
2001-06-01

To grasp the documented evidences available regarding the cause of the anteriorly displaced human temporomandibular joint (TMJ) disc. With 20 specimens of human TMJ from cadavers, at first the number of heads of lateral pterygoid muscle (LPM) bundles were analysed with macroanatomical method. Secondary histological procedure with haematoxyline and eosine staining and orceine staining were carried out for the same specimens. Macroscopically, in 13 specimens an upper and a lower head of the LPM were present. In the other seven specimens, not only an upper and a lower head, but also a third inner head of the LPM covered with a separate fascia was observed. Histochemically, the specific localization of certain bundles of thick elastic fibres in the antero‐medial parts of the anteriorly displaced deformed TMJ disc were observed.
Distribution of mossy fibre rosettes in the cerebellum of cat and mice: evidence for a parasagittal organization at the single fibre level
2001-06-01

Abstract Mossy fibres are the main afferent input to the granular layer of the cerebellar cortex. In this study, the spatial distribution of the mossy fibres' presynaptic enlargements – the so‐called rosettes – were analysed on the single fibre level. Data obtained from the cerebella of cat and mice were compared to look for species differences, and the cerebella of the adult and young mice were also compared to look for developmental changes. The results show that there is a spatial anisotropy in all mossy fibres studied, with neighbouring rosettes being about three times further away from each other along the parasagittal axis and closer to each other in the mediolateral direction. Furthermore, these results suggest that this anisotropy is established at an early developmental stage. The anisotropic orientation of mossy fibres at the single fibre level supports the hypothesis of a timing mechanism in cerebellar function.
Inflammatory Mediators Potentiate ATP-gated Channels through the P2X3 Subunit
2001-06-01

The P2X(3) receptor is an ATP-gated ion channel predominantly expressed in nociceptive neurons from the dorsal root ganglion. P2X(3) receptor channels are highly expressed in sensory neurons and probably contribute to the sensation of pain. Kinetics of P2X(3) currents are characterized by rapid desensitization (<100 ms) and slow recovery (>20 s). Thus, any mechanism modulating rate of desensitization and/or recovery may have profound effect on susceptibility of nociceptive neurons expressing P2X(3) to ATP. Here we show that currents mediated by P2X(3) receptor channels and the heteromeric channel P2X(2/3) composed of P2X(2) and P2X(3) subunits are potentiated by the neuropeptides substance P and bradykinin, which are known to modulate pain perception. The effect is mediated by the respective neuropeptide receptors, can be mimicked by phorbol ester and blocked by inhibitors of protein kinases. Together with data from site-directed mutagenesis our results suggest that inflammatory mediators sensitize nociceptors through phosphorylation of P2X(3) and P2X(2/3) ion channels or associated proteins.
Transgenic Cystic Fibrosis Mice Exhibit Reduced Early Clearance of Pseudomonas aeruginosa from the Respiratory Tract
2001-06-01

Abstract The cystic fibrosis (CF) transmembrane conductance regulator (CFTR) has been proposed to be an epithelial cell receptor for Pseudomonas aeruginosa involved in bacterial internalization and clearance from the lung. We evaluated the role of CFTR in clearing P. aeruginosa from the respiratory tract using transgenic CF mice that carried either the ΔF508 Cftr allele or an allele with a Cftr stop codon (S489X). Intranasal application achieved P. aeruginosa lung infection in inbred C57BL/6 ΔF508 Cftr mice, whereas ΔF508 Cftr and S489X Cftr outbred mice required tracheal application of the inoculum to establish lung infection. CF mice showed significantly less ingestion of LPS-smooth P. aeruginosa by lung cells and significantly greater bacterial lung burdens 4.5 h postinfection than C57BL/6 wild-type mice. Microscopy of infected mouse and rhesus monkey tracheas clearly demonstrated ingestion of P. aeruginosa by epithelial cells in wild-type animals, mostly around injured areas of the epithelium. Desquamating cells loaded with P. aeruginosa could also be seen in these tissues. No difference was found between CF and wild-type mice challenged with an LPS-rough mucoid isolate of P. aeruginosa lacking the CFTR ligand. Thus, transgenic CF mice exhibit decreased clearance of P. aeruginosa and increased bacterial burdens in the lung, substantiating a key role for CFTR-mediated bacterial ingestion in lung clearance of P. aeruginosa.
Processing of Immunosuppressive Pro-TGF-β1,2 by Human Glioblastoma Cells Involves Cytoplasmic and Secreted Furin-Like Proteases
2001-06-01

Abstract TGF-β is a putative mediator of immunosuppression associated with malignant glioma and other types of cancer. Subtilisin-like proprotein convertases such as furin are thought to mediate TGF-β processing. Here we report that human malignant glioma cell lines express furin mRNA and protein, exhibit furin-like protease (FLP) activity, and release active furin into the cell culture supernatant. FLP activity is not modulated by exogenous TGF-β or neutralizing TGF-β Abs. Exposure of LN-18 and T98G glioma cell lines to the furin inhibitor, decanoyl-Arg-Val-Lys-Arg-chloromethylketone, inhibits processing of the TGF-β1 and TGF-β2 precursor molecules and, consequently, the release of mature bioactive TGF-β molecules. Ectopic expression of PDX, a synthetic antitrypsin analog with antifurin activity, in the glioma cells inhibits FLP activity, TGF-β processing, and TGF-β release. Thus, subtilisin-like proprotein convertases may represent a novel target for the immunotherapy of malignant glioma and other cancers or pathological conditions characterized by enhanced TGF-β bioactivity.
Carbohydrate metabolism in ectomycorrhizas: gene expression, monosaccharide transport and metabolic control
2001-06-01

Summary Ectomycorrhizas are mutalistic symbiotic associations formed between fine roots of higher plants, mostly trees, and a wide range of soil ascomycetes and basidiomycetes. It is commonly accepted that there is mutual benefit to the partners, due to the exchange of plant‐derived carbohydrates for amino acids and nutrients supplied by the fungus. While the major concepts of mycorrhizal functioning (exchange of nutrients and metabolites) were proposed in the 1960s, their verification at the molecular level started approximately 10 years ago. This review covers concepts at the molecular level concerned with the fungal carbohydrate supply in symbiosis. We discuss: strategies used by host plants to compensate (and perhaps restrict) carbohydrate drain to the fungal partner; fungal mechanisms that generate strong monosaccharide sinks in colonized plant roots (the formation of a strong carbohydrate sink is a prerequisite for efficient fungal carbohydrate support by the plant partner); and the impact of apoplastic hexose concentration on the regulation of fungal metabolism in symbiosis, since monosaccharides not only serve as nutrients but also as a signal that regulates gene expression.