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Biomedical subjects

M Haas

Publications and source records attributed to M Haas.

At least 91 records · Page 5Linked to original sources
PubMed1998

Restoration of growth arrest by p16INK4, p21WAF1, pRB, and p53 is dependent on the integrity of the endogenous cell-cycle control pathways in human glioblastoma cell lines.

The aim of this study was to demonstrate that the induction of growth arrest in human glioblastoma multiforme (GBM) cell lines by retrovirus-mediated transduction of growth control genes was dependent upon the integrity of specific endogenous control pathways. We assessed the status of the endogenous p16INK4A, p21CIP1, pRb, or p53 genes in eight GBM lines. As expected, we found varied combinations of gene defects. The outcome of transducing five of these cell lines with p16INK4A, p21CIP1, pRb, or p53 genes was not entirely predictable. The growth-inhibitory effects mediated by the transfer of the gene encoding p16 was dependent on the presence of the pRb protein, but was independent of p53 status. p21, a broadly active CDK inhibitor and a strong inducer of growth arrest, was not a universal growth suppressor in the group of glioblastoma cell lines analyzed. The suppression of GBM cell proliferation by viruses encoding pRb or p53 was generally predictable and appeared to be independent of the status of either p16 or p21. Suppression of cell growth was assessed by a colony formation assay, by observance of alterations in morphology, and by cell viability staining for trypan blue exclusion. Our findings suggest that to accomplish the suppression of GBM cell proliferation by the transduction of these cell-cycle control genes, the status of endogenous cell-cycle control genes must be taken into account.

Carrier Proteins↗
PubMed1998

Effect of proteasome inhibitors on monocytic IkappaB-alpha and -beta depletion, NF-kappaB activation, and cytokine production.

We investigated the effect of proteasome inhibitors on the lipopolysaccharide (LPS)-induced expression of several monocytic cytokines, which may be dependent on the transcription factor, nuclear factor-kappaB (NF-kappaB). Exposure of human monocytic THP-1 cells to ALLN and Mu873 prevented the LPS-induced degradation of IkappaB-alpha and -beta, as did the more potent proteasome inhibitor, PSI, whereas several calpain inhibitors were ineffective. This was accompanied by the inhibition of nuclear NF-kappaB binding activity and NF-kappaB transcriptional activation. At the mRNA level, the inhibitors blocked the expression of tumor necrosis factor (TNF) and interleukin-1beta (IL-1beta), whereas IL-8 remained unaffected by ALLN and was only partially reduced by the highest dose of PSI. The latter effect appears to be due to an increase in IL-8 mRNA stability in the presence of proteasome inhibitors. Furthermore, the production of TNF was efficiently suppressed by ALLN and PSI, less by Mu873, and not at all by calpain inhibitors. In primary human blood monocytes ALLN also prevented the LPS-induced degradation of IkappaB-alpha and -beta, efficiently blocked the production of TNF and, to a lesser extent, IL-1beta, whereas that of IL-8 was not inhibited. The expression of NF-kappaB-dependent monocytic cytokines may be selectively controlled by the proteasome, offering a potential therapeutic target in inflammatory disease.

Cell Line↗
PubMed1998

Scavestrogens protect IMR 32 cells from oxidative stress-induced cell death.

Oxidative stress is considered an important pathophysiological mechanism contributing to promote cell death in a broad variety of diseases including cardiovascular and neurodegenerative disorders. The so-called scavestrogens J811 and J861, structurally derived from 17alpha-estradiol, are potent radical scavengers and inhibitors of iron-induced cell damage in vitro. In this study the potential cytoprotective effects of the scavestrogens J811 and J861 against Fenton reagent-induced cell damage (50 microM FeSO4 plus 200 microM H2O2) were compared with those of 17alpha- and 17beta-estradiol. Cell viability studies using Trypan blue staining showed that estradiols and scavestrogens at concentrations ranging from 0.1 to 10 microM are able to protect IMR 32 neuroblastoma cells from Fenton-mediated death. In addition, these compounds decreased lipid peroxidation measured as thiobarbituric acid reactive substances and renormalize oxidative stress-increased intracellular glutathione levels. When given 6 h after the toxic stimulus, J811 and J861 rescued 60% of cells, whereas 17alpha- and 17beta-estradiol were ineffective. These results suggest that the scavestrogens J811 and J861 are powerful antioxidants capable of interfering with radical-mediated cell death in diseases known to be aggravated by reactive oxygen species. Such compounds may be useful in the development of novel treatments for stroke or neurodegenerative disorders.

Antioxidants↗
PubMed1998

Colour vision abnormalities do not correlate with dopaminergic nigrostriatal degeneration in Parkinson's disease.

Sensory disturbances such as olfactory or visual dysfunctions are common in Parkinson's disease (PD). A possible relationship between distorted colour discrimination and the nigrostriatal dopamine deficit is still a matter of debate. We examined 31 de novo Parkinsonian patients with [123I]beta-CIT single photon emission tomography (SPECT). We used a single-head gamma-camera and calculated the binding ratio striatum/cerebellum (specific/nonspecific binding) of [123I]beta-CIT uptake. On the same day, we performed the Farnsworth-Munsell 100 Hue Test (FMT) in these patients and estimated the total error score, in order to investigate abnormalities of colour vision. Parkinsonian patients' total error score was higher compared with an age- and sex-matched control group (P = < 0.0001), whereas disability scores of the Hoehn and Yahr scale (P = 0.019, Spearman r = 0.419) and the Unified Parkinson's Disease Rating Scale (P = 0.039, Spearman r = 0.373) correlated with total error score. No significant association appeared between total error score (Spearman r = -0.119, P = 0.525) and [123I]P-CIT-SPECT ratio. Thus both total error scores of the FMT and [1231]beta-CIT-SPECT binding ratios have been found to reflect the severity of PD. However, only [123I]beta-CIT SPECT reflects degeneration of dopaminergic neurons of the basal ganglia, but does not reflect alterations of the visual system and/or extranigral lesions in PD. From our results, we speculate that FMT may be a valuable clinical method to measure extranigral lesions of the visual system in PD.

Adult↗
PubMed1998

High-titer MSCV-based retrovirus generated in the pCL acute virus packaging system confers sustained gene expression in vivo.

Retroviral gene transfer using vectors encoding tumor suppressor genes has been tested repeatedly as a potential anti-tumor therapy. However, most attempts have been hindered by the inability to deliver genes efficiently and to obtain sustained expression in cells growing in vivo. In this paper we describe a method for producing high-titer MSCV virus using the pCL acute retroviral packaging system. This method facilitates the generation of MSCV virus encoding genes that convey the cytostatic or cytocidal phenotypes of benefit in the treatment of cancer. Amphotropic MSCV virus with an average titer of 6 x 10(6) CFU/ml has been routinely produced in this system. We demonstrate that, unlike the pCL retroviral vectors, the MSCV vector is capable of directing sustained in vivo expression of the green fluorescent protein in infected glioma cells following implantation and tumor growth in nude mice.

Animals↗
PubMed1998

The Na-K-Cl cotransporters.

The Na-K-Cl cotransporters are a class of membrane proteins that transport Na, K, and Cl ions into and out of a wide variety of epithelial and nonepithelial cells. The transport process mediated by Na-K-Cl cotransporters is characterized by electroneutrality (almost always with stoichiometry of 1Na:1K:2Cl) and inhibition by the "loop" diuretics bumetanide, benzmetanide, and furosemide. Presently, two distinct Na-K-Cl cotransporter isoforms have been identified by cDNA cloning and expression; genes encoding these two isoforms are located on different chromosomes and their gene products share approximately 60% amino acid sequence identity. The NKCC1 (CCC1, BSC2) isoform is present in a wide variety of tissues; most epithelia containing NKCC1 are secretory epithelia with the Na-K-Cl cotransporter localized to the basolateral membrane. By contrast, NKCC2 (CCC2, BSC1) is found only in the kidney, localized to the apical membrane of the epithelial cells of the thick ascending limb of Henle's loop and of the macula densa. Mutations in the NKCC2 gene result in Bartter's syndrome, an inherited disease characterized by hypokalemic metabolic alkalosis, hypercalciuria, salt wasting, and volume depletion. The two Na-K-Cl cotransporter isoforms are also part of a superfamily of cation-chloride cotransporters, which includes electroneutral K-Cl and Na-Cl cotransporters. Na-K-Cl cotransporter activity is affected by a large variety of hormonal stimuli as well as by changes in cell volume; in many tissues this regulation (particularly of the NKCCI isoform) occurs through direct phosphorylation/dephosphorylation of the cotransport protein itself though the specific protein kinases involved remain unknown. An important regulator of cotransporter activity in secretory epithelia and other cells as well is intracellular [Cl] ([Cl]i), with a reduction in [Cl]i being the apparent means by which basolateral Na-K-Cl cotransport activity is increased and thus coordinated with that of stimulated apical Cl channels in actively secreting epithelia.

Animals↗
PubMed1998

The effect of different chemotherapeutic agents on the enrichment of DNA mismatch repair-deficient tumour cells.

Loss of DNA mismatch repair is a common finding in hereditary non-polyposis colon cancer as well as in many types of sporadic human tumours. We compared the effect of loss of DNA mismatch repair on drug sensitivity as measured by a clonogenic assay with its effect on the ability of the same drug to enrich for mismatch repair-deficient cells in a proliferating tumour cell population. Mixed populations containing 50% DNA mismatch repair-deficient cells constitutively expressing green fluorescent protein and 50% mismatch repair-proficient cells were exposed to different chemotherapeutic agents. 6-Thioguanine, to which DNA mismatch repair-deficient cells are known to be resistant, was included as a control. The results in the cytotoxicity assays and in the enrichment experiments were concordant. Treatment with either carboplatin, cisplatin, doxorubicin, etoposide or 6-thioguanine resulted in enrichment for mismatch repair-deficient cells, and clonogenic assays demonstrated resistance to these agents, which varied from 1.3- to 4.8-fold. Treatment with melphalan, paclitaxel, perfosfamide or tamoxifen failed to enrich for mismatch repair-deficient cells, and no change in sensitivity to these agents was detected in the clonogenic assays. These results identify the topoisomerase II inhibitors etoposide and doxorubicin as additional agents for which loss of DNA mismatch repair causes drug resistance. The concordance of the results from the two assay systems validates the enrichment assay as a rapid and reliable method for screening for the effect of loss of DNA mismatch repair on sensitivity to additional drugs.

Adenocarcinoma↗
PubMed1998

Renal targeting of a non-steroidal anti-inflammatory drug: effects on renal prostaglandin synthesis in the rat.

1. Renal specific targeting of the non-steroidal anti-inflammatory drug naproxen was obtained by coupling to the low-molecular-mass protein lysozyme. A previous study showed that conjugation to lysozyme resulted in a 70-fold increase of naproxen accumulation in the kidney with a subsequent renal release of the active metabolite naproxen-lysine.2. In the present study we questioned whether naproxen-lysozyme is active in the rat kidney, inhibiting the urinary excretion of prostaglandin E2 and renal sodium and water excretion in salt-restricted baseline conditions as well as during frusemide treatment.3.A high dose of free naproxen (10 mg.day-1. kg-1) did not affect prostaglandin E2 excretion in baseline conditions (naproxen, 11+/-1 ng/8 h; vehicle, 13+/-4 ng/8 h), whereas sodium and water excretion were, respectively, 3.0 and 1.6 times lower in the naproxen group (P<0.05). Naproxen completely prevented the frusemide-induced increase (3-fold) in prostaglandin E2 excretion (naproxen 6.6+/-1.1 ng/8 h, vehicle 40+/-12 ng/8 h, P<0. 005). Frusemide-stimulated natriuresis and diuresis were, respectively, 1.6 (P<0.05) and 1.8 times (P<0.005) lower in the naproxen group.4.A dose of 2 mg.day-1.kg-1 lysozyme-conjugated naproxen did not affect prostaglandin E2 excretion in baseline conditions (conjugate, 18+/-2 ng/8 h; vehicle, 24+/-5 ng/8 h). The conjugate also had no effect on sodium and water excretion. However, the naproxen conjugate completely prevented the frusemide-induced increase (2-fold) in prostaglandin E2 excretion (conjugate, 16+/-3 ng/8 h; vehicle, 48+/-13 ng/8 h, P<0.05). Surprisingly, frusemide-induced natriuresis and diuresis were not affected by the conjugate.5. In conclusion, a renal specific delivery of the non-steroidal anti-inflammatory drug naproxen using lysozyme results in an inhibitory effect on renal prostaglandin E2 synthesis but does not affect the excretion of sodium and water, in contrast to free naproxen.

Animals↗
PubMed1998

Immune complex glomerulonephritis in C4- and C3-deficient mice.

In this study, we examined the roles of C4 and C3 in immune complex glomerulonephritis by actively immunizing C4-deficient (C4 -/-), C3 deficient (C3 -/-) and wild-type mice with apoferritin. Wild-type animals with an intact complement system produced anti-apoferritin IgG and IgM antibodies, and developed mesangial proliferative glomerulonephritis characterized by hypercellularity, matrix expansion, deposition of IgG, IgM, IgA and C3, and the presence of electron dense deposits. In the majority of animals, the peripheral capillaries also contained IgG, C3 and subendothelial and subepithelial electron dense deposits. In contrast to wild-type animals, all apoferritin-immunized C4 -/- and C3 -/- mice had serum cryoprecipitates containing polyclonal IgM and the variable presence of polyclonal IgG. These animals also developed immune complex glomerulonephritis, but their disease manifestations were distinctly different from that of their wild-type littermates. In apoferritin-immunized C4 -/- and C3 -/- mice, IgG was either absent or present in reduced quantities in glomeruli, yet IgM and IgA were present in greater intensity in glomeruli. Capillary wall IgG deposits were absent in all C4 -/- and C3 -/- animals. C4 -/- animals also had significant glomerular C3 deposition, hypercellularity and neutrophil infiltration, which were not present in C3 -/- animals. These results illustrate the complex interplay between the effects of complement to process immune complexes and to lead to inflammation and tissue injury.

Animals↗
PubMed1998

A locus for inherited focal segmental glomerulosclerosis maps to chromosome 19q13.

We performed a genome-wide linkage analysis search for a genetic locus responsible for kidney dysfunction in a large family. This inherited condition, characterized by proteinuria, progressive renal insufficiency, and focal segmental glomerulosclerosis, follows autosomal dominant inheritance. We show with a high degree of certainty (maximum 2-point lod score 12.28) that the gene responsible for this condition is located on chromosome 19q13.

Chromosome Mapping↗
PubMed1998

Funding arrangements for telehealth: encouraging efficiency rather than proliferation.

The use of telehealth as a basis for delivering health services is growing across Australia, and there is clear potential for these technologies to address some of the enduring issues of access and costs of service delivery. However, appropriate incentives must be created to encourage clinicians and managers to evaluate the true opportunity costs and benefits of delivering services in this way against the relevant alternative. This paper examines how different funding arrangements might encourage or discourage efficient use of telehealth.

Australia↗
PubMed1998

Plasma immunadsorption treatment in patients with primary focal and segmental glomerulosclerosis.

BACKGROUND: In primary focal and segmental glomerulosclerosis (FSGS) renal prognosis is poor if no remission of proteinuria can be obtained by treatment. In some patients a permeability factor, responsible for damaging the glomerular epithelial cell and detectable by an in vitro test (GVV-test), seems to be present in the serum. METHOD: We determined the effects of an immunadsorption treatment (IAT) on proteinuria and glomerular permselectivity (using a neutral dextran and dextransulfate-sieving technique to assess glomerular size and charge selectivity) in five patients with FSGS in the native kidneys and three patients with recurrence of FSGS after kidney transplantation. Furthermore, we performed the GVV-test using sera obtained from the patients before and after therapy. RESULTS: IAT reduced proteinuria by more than 50% in four patients, all of whom had an improvement in glomerular-size selectivity. Charge selectivity was better preserved after therapy in three out of these four subjects. The GVV-test prior to IAT was positive in two patients who also responded clinically to therapy. After IAT the GVV-test was negative in all patients, indicating an elimination of the proteinuric factor in the two previously positive patients. CONCLUSION: We conclude that a positive GVV-test before treatment makes a favourable response of IAT on proteinuria likely in patients with FSGS. If a reduction of proteinuria can be obtained by IAT this is due to an improvement in glomerular size and/or charge selectivity.

Adolescent↗
PubMed1998

Taking a count: the evaluation of genetic testing.

While some forms of genetic testing have been available for decades, the progress of the Human Genome Project will expand the possibilities for testing. Evaluation of genetic testing is warranted because health care services have an opportunity cost and thus the benefits of testing must be assessed against the costs. However, genetic testing raises new methodological difficulties in taking into account the full range of costs, benefits and risks. The conventional approach to evaluating new technologies is to assess their benefits in terms of health outcomes only, and to consider the effects on the individuals being tested. Like any test, the product of genetic testing is information. Any subsequent health outcome gain depends on the effectiveness of any intervention which results from the information. Assessing the benefits in terms of health outcomes only excludes consideration of any value, both positive and negative, attached to information. The special feature of genetic testing is that the information obtained has implications for family members. This information may have value to relatives individually and may affect family interactions. Information also has value at a social level; it may affect social relationships and interactions. As the possibilities for genetic testing expand, it is likely that testing programs will be subject to economic evaluation. Until the methods and measures used can validly take this range of effects into account (and into a count of benefits), then the results of evaluation studies will be, at best, incomplete and, at worst, misleading.

Cost-Benefit Analysis↗
PubMed1998

A model of Na-K-2Cl cotransport based on ordered ion binding and glide symmetry.

In the duck red blood cell, Na-K-2Cl cotransport exhibits two modes of ion movement: net cotransport and obligate cation exchange. In high-K cells, the predominant exchange is K/K (or K/Rb). In high-Na cells, it becomes Na/Na (or Na/Li). Both represent partial reactions in which a fully loaded carrier releases part of its cargo, rebinds fresh ions, and returns back across the membrane fully loaded. Net cotransport occurs when the carrier unloads completely and returns empty. This mode has a fixed stoichiometry of 1Na:1K:2Cl under all conditions tested. The ion requirements of the two exchanges differ: K/K exchange requires only K and Cl outside but all three ions inside. Na/Na exchange requires all three ions outside but only Na inside. We propose a simple model in which the carrier can only move when either fully loaded or completely empty and in which the ions bind in a strictly ordered sequence. For example, externally, a Na binds first and then a Cl, followed by a K and a second Cl. Internally, the first on is the first off (glide symmetry), so the Na is released first and then the first Cl, followed by the K and finally by the second Cl. Only then can the empty form return to the outside to start a new cycle.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1998

Angiotensin-converting enzyme gene polymorphism determines the antiproteinuric and systemic hemodynamic effect of enalapril in patients with proteinuric renal disease. Austrian Study Group of the Effects of Enalapril Treatment in Proteinuric Renal Disease.

Angiotensin-converting enzyme (ACE) inhibitors are known to reduce blood pressure and proteinuria in a variety of different glomerular diseases. Nonetheless, a marked interindividual difference in the efficacy of these agents exists. The activity of the ACE and therefore of the renin-angiotensin-aldosterone system (RAAS) has been shown to be under genetic influence. Patients with a deletion genotype at the intron 16 of the ACE gene have been shown to exhibit higher activity of plasmatic ACE when compared to patients with the insertion genotype. We therefore studied prospectively the hemodynamic and antiproteinuric effect of a 6-month therapy with enalapril in patients with biopsy-proven proteinuric glomerular diseases and the DD (n = 10) and ID/II (n = 26) genotype. Although patients with the DD genotype received a slightly higher dose of enalapril, blood pressure and proteinuria did not change significantly. However, both were significantly reduced in the II/ID group after 10 weeks and 6 months of therapy. Creatinine clearance decreased steadily in DD patients. In II/ID patients, creatinine clearance was reduced significantly after 10 weeks of therapy but increased again thereafter and the value at 6 months was again comparable to the one obtained in the DD patients. We conclude from our study that the ACE genotype influences the blood pressure-lowering and antiproteinuric effect of enalapril in patients with proteinuric glomerular disease.

Adult↗
PubMed1998

Drug delivery to the kidneys and the bladder with the low molecular weight protein lysozyme.

The low molecular weight protein (LMWP) lysozyme is a suitable drug carrier for renal drug targeting. When the tubular reabsorption of a LMWP can be prevented, the protein will be excreted in the urine. In this way, lysozyme (LZM) conjugates might also be used as carriers for targeting to the urinary tract. Since positive domains on the protein surface are important for the interaction with the tubular uptake-receptor, we studied the urinary excretion of a drug-LZM conjugate with and without positive charge on the LMWP. We synthesized two conjugates with the fluorescent compound fluorescein. A positively charged conjugate was obtained by reacting fluorescein isothiocyanate (FITC) with LZM at a 1:1 molar to molar ratio; this conjugate contained six free primary aminogroups. The conjugate without positively charged groups was obtained by reacting the remaining free primary aminogroups of the FITC-LZM with succinic anhydride (Suc). The Suc-FITC-LZM contained only 0.2 free primary aminogroups per molecule. We studied the pharmacokinetics of the conjugates in freely moving Wistar rats. The FITC-LZM conjugate was excreted intactly into the urine for 29 +/- 4% of the injected dose. The Suc-FITC-LZM was excreted into the urine intactly for 45 +/- 4%. These data indicate that the excretion of a drug-LMWP conjugate into the urine can be increased by decreasing the positive charge on the carrier surface. Such a carrier may be an attractive candidate for drug targeting to the bladder.

Animals↗