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K Németh

Publications and source records attributed to K Németh.

33 records · Page 2Linked to original sources
PubMed1993

Immunocytochemical visualization of kappa-opioid receptors on chick embryonic neurons differentiating in vitro.

The present paper is the first immunocytochemical demonstration of kappa-opioid receptors in neurons isolated from seven-day-old chick embryonic forebrains and cultivated for one to seven days. The monoclonal antibody KA8 (IgG1-k) [Maderspach et al. (1991) J. Neurochem. 56, 1897-1904] was raised against the frog brain kappa-opioid receptor as an antigen and recognizes an epitope in or near the ligand binding site. The KA8 immunostaining of the neurons displayed individual variations and changed with the in vitro differentiation. Receptors often appeared at the pole of the primary outgrowing process, later on in the whole soma and finally on the branched processes. Specific radioligand binding and KA8 immunocytochemistry both presented an increase in the receptor concentration with development. The equilibrium binding values that were measured at 1 nM [3H]naloxone concentration were 2.9 and 6.1 fmol/10(6) cells on the first and sixth cultivation days, respectively. Neurons were treated with 10(-7) M bremazocine or dynorphine (agonists with relative specificity to kappa-opioid receptors) on the second and third cultivation days. The agonist promoted the morphological differentiation which was already visible within 24 h. It also promoted the expression of the 200,000 mol. wt neurofilament protein, this became pronounced after two to three days. The changes provoked by the agonist were reduced by the opioid antagonist norbinaltorphimine (10(-7) M) or naloxone (10(-5) M) indicating that the effect was receptor-mediated. The hypothesis that kappa-opioid agonists through their receptors may function as regulatory signals in the early neuronal differentiation is discussed.

Animals↗
PubMed1992

[Detection of delta F508 mutation in cystic fibrosis].

The common major mutation (delta F508) resulting in the removal of a phenylalanine residue of the cystic fibrosis gene product has been identified in patients with cystic fibrosis in several European countries. The frequency of this mutation was analyzed in 39 Hungarian patients with cystic fibrosis and in some of their relatives. In 43 out of the total 65 persons involved in the study (66.15%), and in 50% of cystic fibrosis chromosomes the delta F508 mutation could be detected. 56.4% of the patients were homozygous for the mutation. The frequency of the delta F508 mutation in these Hungarian patients resembles the values found in Middle Europe. Pulmonary and enteral symptoms were more severe in cystic fibrosis patients with delta F508 mutation. Although the number of patients is relatively low, missing or mild lung involvement characterized the cases with non-delta F508 mutation.

Adult↗
PubMed1992

Differential Expression of Histone H3 Gene Variants during Cell Cycle and Somatic Embryogenesis in Alfalfa.

Northern analysis has revealed substantial differences in mRNA accumulation of the two histone H3 gene variants represented by pH3c-1 and pH3c-11 cDNA clones. Both in partially synchronized cell suspension cultures and in protoplast-derived cells from alfalfa, Medicago varia, the maximal level of the histone H3-1 gene transcript coincided with the peak in [(3)H]thymidine incorporation. Histone H3-11 mRNA was detectable in cells throughout the period of the cell cycle studied. Various stress factors such as medium replacement, enzyme digestion of the cell wall, osmotic shock, and auxin treatment considerably increased the level of the histone H3-11 transcript. In alfalfa (Medicago sativa), the presence of H3-11 mRNA in unorganized tissues of microcallus suspension and in somatic embryos induced by auxin treatment supports the idea that this H3 variant exists in a continously active state of transcription. During embryo development, the early globular stage embryos showed increased accumulation of histone H3-11 mRNA in comparison with the later stages. The highest level of the histone H3-1 transcript was detectable 1 day after treatment of callus tissues with 2,4-dichlorophenoxyacetic acid. Somatic embryos contained appreciable levels of histone H3-1 transcripts at all stages of somatic embryo development. These observations suggest that the histone H3-1 gene belòngs to the class of replication-dependent histone genes. The histone H3-11 gene showed characteristics of a constitutively expressed replacement-type histone gene, with a specific characteristic that external factors can influence the level of gene transcription.

Journal Article↗
PubMed1991

Complementation of a yeast cell cycle mutant by an alfalfa cDNA encoding a protein kinase homologous to p34cdc2.

The cdc2 protein kinase plays a central role in control of the eukaryotic cell cycle of animals and yeasts. We have isolated a cDNA clone (cdc2Ms) from alfalfa (Medicago sativa L.) that is homologous to the yeast cdc2/CDC28 genes. The encoded protein is 64% identical to the yeast and mammalian counterparts and shows all the prominent structural features known from these organisms. Antibody raised against a 16-amino acid synthetic peptide with crossreactivity against p34 proteins recognized a 34-kilodalton protein in extracts of alfalfa cells. When transferred into a fission yeast, the plant cdc2 homolog can complement a temperature-sensitive cdc2 mutant. Northern analysis revealed higher transcript levels in shoots and suspension cultures than in roots. In addition to the dominant transcript of 1.4 kilobases detected in the poly(A)+fraction, 2.5- and 1.2-kilobase transcripts were detected in total RNA preparations from shoots or somatic embryos. Suspension cultures that were induced to form somatic embryos by an auxin (2,4-dichlorophenoxyacetic acid) showed fluctuations in transcription pattern during the induction period and embryogenesis.

Amino Acid Sequence↗
PubMed1991

Alfalfa heat shock genes are differentially expressed during somatic embryogenesis.

We have isolated two cDNA clones (Mshsp18-1; Mshsp18-2) from alfalfa (Medicago sativa L.) which encode for small heat shock proteins (HSPs) belonging to the hsp17 subfamily. The predicted amino acid sequences of the two alfalfa proteins are 92% identical and a similar degree of homology (90%) can be detected between Mshsp18-2 and the pea hsp17. In comparison to various members of small HSPs from soybean amino acid sequence similarities of 80-86% were identified. The alfalfa HSPs share a homologous stretch of amino acids in the carboxy terminal region with hsp22, 23, 26 from Drosophila. This region contains the GVLTV motif which is characteristic of several members of small HSPs. At room temperature alfalfa hsp18 mRNAs were not detectable in root and leaf tissues but northern analysis showed a low level of expression in microcallus suspension (MCS). The transcription of Mshsp18 genes is induced by elevated temperature, CdCl2 treatment and osmotic shock in cultured cells. In alfalfa somatic embryos derived from MCS a considerable amount of hsp18 mRNA can be detected during the early embryogenic stages under normal culture conditions. The differential expression of these genes during embryo development suggests a specific functional role for HSPs in plant cells at the time of the developmental switch in vitro.

Amino Acid Sequence↗
PubMed1991

A monoclonal antibody recognizing kappa- but not mu- and delta-opioid receptors.

A monoclonal antibody (mAb), KA8 that interacts with the kappa-opioid receptor binding site was generated. BALB/c female mice were immunized with a partially purified kappa-opioid receptor preparation from frog brain. Spleen cells were hybridized with SP2/0AG8 myeloma cells. The antibody-producing hybridomas were screened for competition with opioid ligands in a modified enzyme-linked immunosorbent assay. The cell line KA8 secretes an IgG1 (kappa-light chain) immunoglobulin. The mAb KA8 purified by affinity chromatography on protein A-Sepharose CL4B was able to precipitate the antigen from a solubilized and affinity-purified frog brain kappa-opioid receptor preparation. In competition studies, the mAb KA8 decreased specific [3H]ethylketocyclazocine ([3H]EKC) binding to the frog brain membrane fraction in a concentration-dependent manner to a maximum to 72%. The degree of the inhibition was increased to 86% when mu- and delta-opioid binding was suppressed by 100 nM [D-Ala2,NMe-Phe4,Gly-ol]-enkephalin (DAGO) and 100 nM [D-Ala2,L-Leu5]-enkephalin (DADLE), respectively, and to 100% when mu-, delta-, and kappa 2-sites were blocked by 5 microM DADLE. However, the mu-specific [3H]DAGO and the delta-preferring [3H]DADLE binding to frog brain membranes cannot be inhibited by mAb KA8. These data suggest that this mAb is recognizing the kappa- but not the mu- and delta-subtype of opioid receptors. The mAb KA8 also inhibits specific [3H]naloxone and [3H]EKC binding to chick brain cultured neurons and rat brain membranes, whereas it has only a slight effect on [3H]EKC binding to guinea pig cerebellar membranes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

In vitro and in vivo (LD50) effects of human lactoferrin on bacteria.

The in vitro and in vivo effects of human lactoferrin (LF), apoLF, iron saturated LF and of different iron containing compounds (ferric chloride, ferric sodium citrate) were studied on Escherichia coli, Salmonella typhi-murium and Pseudomonas aeruginosa reference and wild-type strains with well-defined virulence markers (i.e. enterochelin, aerobactin production). LF exert in vitro antibacterial effect, and iron-free Vogel-Bonner medium proved to be suitable for its determination. The effect of intraperitoneally administered LF could not be evaluated because of its aspecificity, as any treatment (e.g. saline, Ringer solution) before bacterial challenge activated macrophages. In contrast to the in vitro results, intramuscular challenge failed to inhibit bacterial growth in vivo, as siderophores produced by bacteria were able to acquire lactoferrin-bound iron. LF treatment, like iron addition, enhanced the virulence of bacteria in mice, whereas apoLF - using iron present in the body fluids - turned to LF being unable to acquire siderophore-bound iron from bacteria. These findings do not support the literary view that LF would be useful as an antimicrobial drug.

Animals↗
PubMed1985

Induction of human rheumatoid factor and other autoantibodies by bacterial lipopolysaccharide.

In vitro autoantibody production induced by lipopolysaccharide (LPS) was studied using peripheral blood mononuclear (PBM) cell suspensions from patients with rheumatoid arthritis (RA) and healthy subjects. PBMs from both groups could be induced by LPS to secrete IgM and IgA rheumatoid factors (RF), antinuclear and anti-beta-2-microglobulin autoantibodies. Spontaneous production of IgM-RF was considerably higher in RA than in controls. The rate of IgM-RF and IgA-RF secretion detected by ELISA increased with the dose of LPS in cultures of both groups. In RA, differences were found between the kinetics of IgM- and IgA-RFs secretion. LPS augmented the relative avidity of IgM-RF produced by PBMs from RA patients and this value was significantly higher than that of healthy persons. In some cases RFs cross-reacting with nuclear antigens and beta-2-microglobulin were detected.

Antibodies, Antinuclear↗
PubMed1982

[Iron-deficiency and replacement in infancy].

The frequency of iron-deficiency anaemia was investigated in 376 children with the aid of serum ferritin determinations. The children, resident in Budapest or Miskolc were 6 months to 3 years old. Marked iron-deficiency (serum ferritin less than 10 micrograms/l) could be demonstrated in 33.8% of the investigated population. The frequency of iron-deficiency was related to the socio-economic and cultural status of the mother, a factor which is thought to influence the feeding habits. In areas where iron-deficiency is frequently detected regular iron-prophylaxis is recommended.

Anemia, Hypochromic↗