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Biomedical subjects

A Werner

Publications and source records attributed to A Werner.

At least 109 records · Page 6Linked to original sources
PubMed1998

Intercellular adhesion molecule-1 (ICAM-1) in the mouse facial motor nucleus after axonal injury and during regeneration.

Intercellular adhesion molecule 1 (ICAM-1, CD54) is a widely expressed glycoprotein, which plays an important role in leukocyte extravasation and in the interaction of lymphocytes with antigen-presenting cells. In the current study we examined the regulation of ICAM-1 in the mouse facial motor nucleus after facial nerve transection, using immunohistochemistry, confocal laser microscopy and electron microscopy. In the normal facial nucleus ICAM-1 immunoreactivity was restricted to vascular endothelium. Transection of the facial nerve led to a strong and selective upregulation of ICAM-1 on activated microglia. Quantitation of microglial ICAM-1 immunoreactivity revealed a biphasic increase. The first peak 1-2 days post operation paralleling the early stage of microglial activation was followed by a decline at 4-7 days. The second induction of ICAM-1 occurred at day 14 accompanying the period of neuronal cell death and microglial phagocytosis of neuronal debris. Immunoelectron microscopy showed strong ICAM-1 reactivity on the cell membrane of activated microglia at day 2. During the second peak (day 14), ICAM-1 was also observed on lymphocytes adhering to phagocytotic microglia forming aggregates around neuronal debris. No immunolabelling was observed on neurons, astrocytes or oligodendroglia. These data suggest the involvement of ICAM-1 in the adhesion of activated microglia, in their phagocytosis of neuronal debris, and also in the interaction with infiltrating lymphocytes following this injury.

Animals↗
PubMed1998

Improved nuclear magnetic resonance microscopic visualization of joint cartilage using liposome entrapped contrast agents.

RATIONALE AND OBJECTIVES: After intraarticular application of gadolinium (Gd)-DTPA the visualization cartilage surface roughness is limited because of diffusion into the cartilage. To improve the sensitivity of magnetic resonance (MR) arthrography to diagnose cartilage surface abnormalities, the authors have tested liposome-entrapped contrast agents. METHODS: Using paramagnetic contrast agents (Gd-DTPA and manganese chloride) free and entrapped in liposomes, respectively, high resolution MR imaging investigations were performed at 7.1 tesla on intact pig temporomandibular and rabbit knee joints. RESULTS: After intraarticular injection of the liposome-entrapped contrast agents an excellent contrast between cartilage surface and joint space was achieved. Diffusion of the contrast agent into the cartilage layer was prevented and the visualization of the cartilage surface was improved markedly. Small mechanically and enzymatically induced cartilage lesions could be assessed reliably. CONCLUSIONS: Intraarticular injection of liposome-entrapped contrast agents can improve the potential of MR arthrography concerning the detection of early osteoarthritic cartilage changes.

Animals↗
PubMed1998

Na-P(i) cotransport sites in proximal tubule and collecting tubule of winter flounder (Pleuronectes americanus).

Localization of a recently described and cloned Na-Pi cotransport system from flounder was investigated by reverse transcription-polymerase chain reaction (RT-PCR) of microdissected tubules and by immunocytochemistry of kidney of winter flounder. Histological examination showed a small glomerulus, an extremely short proximal tubule PI with a selective affinity to Lens culinaris agglutinin from lentils, and an extensive second proximal tubule segment PII (> 90% of proximal tubules), consisting of cells with numerous apical clear vesicles and extensive amplification of basolateral cell membranes. PII merged with the collecting tubule/ collecting duct (CT/CD) system without a distal segment. By RT-PCR, PII cells revealed high levels of NaPi-II related RNA; low levels were also observed in CTs. Previously characterized antisera against different epitopes of flounder NaPi-II specifically labeled the basolateral regions of PII and the apical cell portion of CT/CD cells and of some PII cells. These results suggest that tubular secretion of P(i) occurs in PII of teleost fish with modulation of urinary P(i) content in the subsequent CT/CD system.

Animals↗
PubMed1998

Increased cytosolic Ca2+ amplifies oxygen radical-induced alterations of the ultrastructure and the energy metabolism of isolated rat pancreatic acinar cells.

BACKGROUND: Oxygen radicals have been implicated as important mediators in the early pathogenesis of acute pancreatitis, but the mechanism by which they produce pancreatic tissue injury remains unclear. We have, therefore, investigated the effects of oxygen radicals on isolated rat pancreatic acinar cells as to the ultrastructure, cytosolic Ca2+ concentration and energy metabolism. METHODS: Acinar cells were exposed to an oxygen radical-generating system consisting of xanthine oxidase, hypoxanthine and chelated iron ions. Cell injury was assessed by LDH release and electron microscopy. Cytosolic Ca2+ levels and mitochondrial membrane potential were determined by flow cytometry; adenine nucleotide concentrations by HPLC. Mitochondrial dehydrogenase activity was measured by spectrophotometric assay. RESULTS: Oxygen radicals damaged the plasma membrane as shown by a 6-fold LDH increase in the incubation medium within 180 min. At the ultrastructural level, mitochondria were the most susceptible to oxidative stress. In correlation to the pronounced mitochondrial damage, the mitochondrial dehydrogenase activity declined by 70%, whereas the mitochondrial membrane potential was enhanced by 27% after 120 min. Together this may cause the 85% decrease in the ATP concentration and the corresponding increase in ADP/AMP observed in parallel. In addition, an immediate 26% increase in cytosolic Ca2+ was found, a change which could be inhibited by BAPTA, reducing cellular damage. CONCLUSION: Cytosolic Ca2+ synergizes with oxygen radicals causing alterations of the ultrastructure and energy metabolism of acinar cells which might contribute to the cellular changes found in early stages of acute pancreatitis.

Acute Disease↗
PubMed1998

Chemoattractant factors and the control of human immunodeficiency virus replication.

Factors secreted by CD8(+) T cells have been described to suppress immunodeficiency virus replication. The research efforts to identify these factors led to the proposal of some candidate proteins as being responsible for the antiviral effects. Chemokines and IL-16 are secreted by CD8(+) T cells and inhibit HIV replication through different mechanisms. However, their antiviral properties cannot fully explain the inhibitory activities found in cell culture supernatants from CD8(+) T cells.

Antiviral Agents↗
PubMed1998

Structural integrity of resin-modified glass ionomers as affected by the delay or omission of light activation.

Since light activation of resin-modified glass ionomers as a means of polymerizing the HEMA is usually done shortly after mixing occurs, the acid-base reaction will proceed mainly within a formed HEMA-polymer matrix. Delaying or omitting light activation may alter the structure and consequently its integrity. The aim of this study was to investigate the effect on the structural integrity of Fuji II LC, Photac-Fil, and Vitremer by delaying or omitting light initiation as compared with the integrity when light activation is performed 2 min after mixing occurs. We evaluated integrity by three-body wear experiments, conducted 8 hrs after sample preparation, to establish the integrity in the early phase of hardening, as well as after 1 wk and after 4 mos, to follow the materials throughout the process of maturation. When light activation was delayed for 1 hr, the structural integrity of Fuji II LC and Photac-Fil improved significantly in the early stages of hardening. In the case of Vitremer, an hour's delay of light activation significantly decreased integrity, which declined further when light activation was omitted. Fuji II LC was not affected by the omission of light activation, while Photac-Fil was markedly weakened. After 4 mos of aging, most of the samples of each product which had been cured by the different methods attained equal integrity, with the exception of the non-light-activated Vitremer samples, which remained weaker. We concluded that the structural integrity of resin-modified glass-ionomer cements benefits from a chemical integration of the polyalkenoate and poly-HEMA networks, as in Vitremer. Improvement in the structural integrity in the early phase for cements with a mechanical entanglement of the matrices, as in Fuji II LC and Photac-Fil, requires an acid-base reaction, a considerable portion of which may take place before activation of the HEMA polymerization.

Composite Resins↗
PubMed1998

Na+-dependent phosphate cotransporters: the NaPi protein families.

In vertebrates, the level of inorganic phosphate (Pi) is tightly balanced both inside the cell and in the whole organism. A number of different Na+-dependent Pi cotransport systems involved in Pi homeostasis have been identified and characterized at the molecular level in the past 7 years. The transporters constitute three different protein families denoted NaPi-I, NaPi-II and NaPi-III. NaPi-I from the rabbit was the first member of this family to be cloned. However, it still resists efforts to unravel its physiological role and a clear-cut functional identity: is it a Cl- channel, a Na+/Pi cotransporter, a regulator, or does it perform a combination of these functions? These questions provide a slight taste of the problems associated with orphan genes derived from sequencing projects. The members of the NaPi-II protein family are crucially involved in tightly controlled renal Pi excretion and, as recently discovered, intestinal Pi absorption. The expression and the cellular distribution of NaPi-II in the proximal tubular epithelium are affected by hormonal and metabolic factors known to influence extracellular fluid Pi homeostasis. Recently, the expression of NaPi-II has been demonstrated in osteoclasts and brain; however, the physiological roles of NaPi-II in these tissues remain to be established. The members of the third protein family, NaPi-III, have been identified on the basis of their function as viral receptors. The widespread expression of this family suggests that NaPi-III is involved in supplying the basic cellular metabolic needs for Pi.

Animals↗
PubMed1998

[B-technique after augmentation mammaplasty to correct dislocation of nipple and areola].

"B-technique" was first developed for breast reduction. In the past there were some new information characterizing the "B-technique" as being a good method for some breast cancer patients undergoing conversation surgery as well. In this paper we report on a patient suffering from anisomastia; after augmentation mammaplasty the "B-technique" was used to put the nipple and areola in the right position. If the dislocation of the nipple is not extreme we considered the "B-technique" a favourable method to obtain a good aesthetic result.

Adolescent↗
PubMed1997

The biochip. A new membrane bioreactor system for the cultivation of animal cells in defined tissue-like cell densities.

Based on the laminar structure of the human liver tissue, a high cell density membrane bioreactor was developed that emulates a cell layer thickness of 40 microns. The "biochip" consists of a platinum-coated metal cell grid covered with two microfiltration membranes to form separate cell chambers of defined volume. Starting with a continuous chemostat process, the viability of a model suspension cell culture could be stabilized at 98%. In a second step these cells were transferred into the biochip system and were cultivated successfully for several days under tissue-like cell densities in a modified membrane holder under cross-flow conditions.

Animals↗
PubMed1997

Expression cloning and characterization of a renal organic anion transporter from winter flounder.

The cDNA coding for a renal p-aminohippurate (PAH) transporter from winter flounder (Pseudopleuronectes americanus), designated fROAT, was cloned by functional expression in Xenopus laevis oocytes. fROAT is approximately 2.8 kbp in length and encodes a protein of 562 amino acids, related to the rat renal organic anion transporter ROAT1/OAT1 and the organic cation transporters OCT1 and OCT2. In oocytes, fROAT mediated probenecid-sensitive PAH uptake, with a Km for PAH of about 20 microM, and inhibited by external glutarate (GA) (1 mM). The functional characteristics suggest that fROAT is the basolateral PAH/dicarboxylate exchanger of the flounder kidney.

Amino Acid Sequence↗
PubMed1997

Electrophysiological characterization of the flounder type II Na+/Pi cotransporter (NaPi-5) expressed in Xenopus laevis oocytes.

The two electrode voltage clamp technique was used to investigate the steady-state and presteady-state kinetic properties of the type II Na+/Pi cotransporter NaPi-5, cloned from the kidney of winter flounder (Pseudopleuronectes americanus) and expressed in Xenopus laevis oocytes. Steady-state Pi-induced currents had a voltage-independent apparent K(m) for Pi of 0.03 mM and a Hill coefficient of 1.0 at neutral pH, when superfusing with 96 mM Na+. The apparent K(m) for Na+ at 1 mM Pi was strongly voltage dependent (increasing from 32 mM at -70 mV to 77 mM at -30 mV) and the Hill coefficient was between 1 and 2, indicating cooperative binding of more than one Na+ ion. The maximum steady-state current was pH dependent, diminishing by 50% or more for a change from pH 7.8 to pH 6.3. Voltage jumps elicited presteady-state relaxations in the presence of 96 mM Na+ which were suppressed at saturating Pi (1 mM). Relaxations were absent in non-injected oocytes. Charge was balanced for equal positive and negative steps, saturated at extremes of potential and reversed at the holding potential. Fitting the charge transfer to a Boltzmann relationship typically gave a midpoint voltage (V0.5) close to zero and an apparent valency of approximately 0.6. The maximum steady-state transport rate correlated linearly with the maximum Pi-suppressed charge movement, indicating that the relaxations were NaPi-5-specific. The apparent transporter turnover was estimated as 35 sec-1. The voltage dependence of the relaxations was Pi-independent, whereas changes in Na+ shifted V0.5 to -60 mV at 25 mM Na+. Protons suppressed relaxations but contributed to no detectable charge movement in zero external Na+. The voltage dependent presteady-state behavior of NaPi-5 could be described by a 3 state model in which the partial reactions involving reorientation of the unloaded carrier and binding of Na+ contribute to transmembrane charge movement.

Animals↗
PubMed1997

Molecular cloning, sequence, expression, and processing of the interleukin 16 precursor.

Interleukin 16 (IL-16) has been shown to function as chemoattractant factor, as a modulator of T-cell activation, and as an inhibitor of immunodeficiency virus replication. The recent identification of inconsistencies in published IL-16 cDNA nucleotide sequences led to the proposal that IL-16 is synthesized in the form of a large precursor protein (pro-IL-16). To identify the true transcriptional start of the IL-16 mRNA rapid amplification of cDNA ends methods were applied. The complete pro-IL-16 cDNA was subsequently molecularly cloned, sequenced, and expressed in COS-7 cells. We report here that pro-IL-16 is most likely synthesized as a 67-kDa protein and is encoded from a major 2.6-kb transcript. Recombinant pro-IL-16 polypeptides are specifically cleaved in lysates of CD8(+) cells, suggesting that the naturally secreted bioactive form of IL-16 is smaller than the originally published 130 amino acids fragment. Moreover, in contrast to other interleukins such as IL-15, IL-16 mRNA expression is almost exclusively limited to lymphatic tissues underlining the potential of IL-16 as an immune regulatory molecule.

Amino Acid Sequence↗
PubMed1997

Human connective tissue growth factor is expressed in advanced atherosclerotic lesions.

BACKGROUND: Atherosclerosis affects certain but not all vascular beds of the human circulation. Its molecular mechanisms are only partially understood. Human connective tissue growth factor (hCTGF) is a novel cysteine-rich, secreted polypeptide. hCTGF is implicated in connective tissue formation, which may play an important role in atherosclerosis. METHODS AND RESULTS: By using a differential cloning technique, we isolated a cDNA clone from a human aorta cDNA library, which is identical to hCTGF. Northern analysis shows that hCTGF mRNA was expressed at 50- to 100-fold higher levels in atherosclerotic blood vessels compared with normal arteries. In vascular smooth muscle cells, high-level expression of hCTGF mRNA was induced by transforming growth factor-beta 1. Using in situ hybridization and immunohistochemistry, we found that all advanced atherosclerotic lesions of human carotid arteries (eight patients; mean age, 69; age range, 57 to 85 years) and femoral arteries (two patients; mean age, 71.5 years) that we tested expressed high levels of both hCTGF mRNA and protein. hCTGF expression was localized mainly to smooth muscle cells in the plaque lesions that are negative for proliferating cell nuclear antigen staining. In addition, some CD-31-positive endothelial cells of plaque vessels expressed high levels of hCTGF mRNA and protein. hCTGF-positive cells were found predominantly in areas with extracellular matrix accumulation and fibrosis. In contrast, in normal arteries, we were unable to detect either hCTGF mRNA or immunoreactive hCTGF protein. CONCLUSIONS: In the present study, we have shown for the first time that both hCTGF mRNA and protein are expressed in human arteries in vivo and that hCTGF may represent a novel factor expressed at high levels specifically in advanced lesions and may play a role in the development and progression of atherosclerosis.

Adult↗
PubMed1997

T cell repertoire in patients with multiple myeloma and monoclonal gammopathy of undetermined significance: clonal CD8+ T cell expansions are found preferentially in patients with a low tumor burden.

The T cell receptor (TCR) variable (V) gene repertoire was analyzed in patients with monoclonal gammopathy of undetermined significance (MGUS) (n = 17), multiple myeloma (MM) stage I (n = 16), MM stages II/III (n = 31) and age-matched controls (n = 27) by immunofluorescence and flow cytometry using a panel of mouse monoclonal antibodies (mAb) (n = 10) against TCR V alpha and V beta gene products. T cell expansion was defined as a value > or = thrice the normal median value for each respective TCR V mAb. Fifty-three percent of all patients displayed CD8+ expansion(s) as compared to 30% of age-matched controls (p < 0.001). Within the CD4 subset, 18% of the patients displayed T cell expansion(s) in comparison to 11% of the controls (not significant). Interestingly, the CD8+ expansion(s) were more frequently noted in patients with a low tumor burden (MGUS/MMI) (73%) as compared to those with advanced disease (MM II/III) (32% and control donors (30%) (p < 0.01). Likewise, multiple CD8+ expansions (two or more) were more common in MGUS/MM I patients than in MM II/III and controls (p < 0.01). The T cell expansions were stable over time in patients with a stable disease. A high degree of clonality of the expansions was detected by TCR CDR3 fragment length analysis, determination of J beta gene usage and nucleotide sequencing. The frequent finding of oligoclonal CD8+ T cell expansions in patients with a low tumor mass, but not in patients with advanced disease justifies further work in order to identify the relevance of expanded CD8+ T cells. In one patient with T cell reactivity against the autologous myeloma idiotype, two expansions within the CD8 population (V beta 3 and V beta 5.2 respectively) displayed no reactivity against the idiotype. Instead, idiotype recognition was confined to a CD8 non-expanded V beta 22+ T cell population, with a highly restricted TCR usage (CDR3 fragment length analysis).

Adult↗
PubMed1997

Distribution of heat-shock protein 60 immunoreactivity in testes of infertile men.

The immunohistochemical localization of heat-shock protein 60 (hsp60) was investigated in testicular biopsies obtained from 121 adult men with disturbed fertility. In normal unaffected tubules, hsp60 immunoreactivity was localized to spermatogonia, primary spermatocytes and Sertoli cells. In spermatogonia, cytosolic and mitochondrial labelling could be differentiated. In general, the number of stained spermatogonia decreased with the loss of spermatogenic function. A significant (P<0.01) reduction of stained spermatogonia was observed in testes with maturation arrest of spermatogenesis at the level of primary spermatocytes (30.2+/-21.6%) compared with testes exhibiting normal spermatogenesis. In addition, the decrease in the score correlated significantly with the diminution of cytosolic hsp60 immmunolabelling (coefficient r=0.25, P=0.03). There was a significant difference (P<0.01) in the percentage of cytosolic-stained spermatogonia in testes with a score equal to or greater than 5 (14.7+/-9.8%) and a score less than 5 (8. 9+/-6.9%). These observations suggest that a low level of hsp60 expression in spermatogonia may lead to a different pattern of protection, which in turn could be involved in low spermatogenic efficiency.

Adult↗
PubMed1997

Clonal CD8+ and CD52- T cells are induced in responding B cell lymphoma patients treated with Campath-1H (anti-CD52).

Five patients with non-Hodgkin's lymphoma (NHL) and 4 patients with chronic lymphocytic leukaemia (CLL) were treated with the CDR-grafted (rat x human) monoclonal antibody (mAb) Campath-1H (anti-CD52). Tumour regression was noted preferentially in peripheral blood and in the bone marrow but lymph nodes were less affected. Normal blood B and T cells were profoundly reduced in all patients whereas CD16+ NK cells and CD14+ monocytes decreased marginally. In all responding CLL patients CD52-negative T but not B cells appeared during treatment and persisted for several months (4-19+) during unmaintained follow-up. Clonal T cells defined as a predominance of a single T cell receptor (TCR) V gene usage, in one case verified by TCR CDR3 fragment analysis and nucleotide sequencing, emerged within the CD52-/CD8+ cell population during Campath-1H therapy in 2 CLL patients, both achieving a long-lasting remission. The increase in CD8+ T cell expansions (up to 23-fold) during unmaintained remission and follow-up suggest that the clonal CD8+ cells may represent regulatory T cells controlling the growth of the tumour B cell clone. Clonal T cells might thus be a target for an immune therapeutic intervention in B cell tumours.

Adult↗
PubMed1997

Increased serum and mRNA levels of RANTES associated with elevated levels of activated CD8+CD38+ T cells in HIV-1 infected individuals.

The serum levels of the beta-chemokine RANTES and, albeit less, MIP-1 beta were found to be increased in 37 HIV-1 infected compared to seronegative individuals. In contrast the serum levels of IL-16 were only sporadically elevated in seropositives as well as in seronegatives. Concomitantly, the RANTES gene expression increased about tenfold in seropositives, whereas the MIP-1 beta and IL-16 mRNA levels were not elevated. No correlation between the increase of the MIP-1 beta and RANTES serum concentrations and the plasma virus load, the number of the peripheral CD4+ T cells or the therapy status of the patients was found. However, the increased proportion of activated CD8+CD38+ T cells in the peripheral blood of all seropositives paralleled the increased RANTES serum levels detected indicating that immune activation in HIV-1-infected individuals may contribute to increased RANTES serum levels.

ADP-ribosyl Cyclase↗