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Biomedical subjects

B Blum

Publications and source records attributed to B Blum.

At least 19 recordsLinked to original sources
PubMed1992

Formation of guide RNA/messenger RNA chimeric molecules in vitro, the initial step of RNA editing, is dependent on an anchor sequence.

Synthetic pre-edited messenger RNA (mRNA) and guide RNA (gRNA) for the 5'-edited maxicircle-encoded ND7 cryptogene from Leishmania tarentolae formed chimeric molecules upon incubation in the presence of a mitochondrial extract. These chimeric molecules consisted of the gRNAs covalently linked to the mRNAs by short oligo(U) tails at normal editing sites in most cases. Unlike the previously reported chimeras present in steady-state kinetoplast RNA, the in vitro-synthesized chimeras showed no editing of downstream editing sites. The synthesis of chimeric RNAs required ATP and was dependent on the formation of a gRNA/mRNA anchor duplex 3' of the pre-edited region, as shown by in vitro mutagenesis of the mRNA and the gRNA. mRNA sequences 3' and 5' of the pre-edited region also affected the efficiency of the chimera-forming activity. This in vitro system may accurately represent the initial step in RNA editing.

Animals↗
PubMed1992

Generation of unexpected editing patterns in Leishmania tarentolae mitochondrial mRNAs: misediting produced by misguiding.

We have analyzed the generation of unexpected patterns of RNA editing, i.e., those not following a strict 3' to 5' progression, which occur in junction regions between fully edited and preedited sequences. Evidence is presented that these patterns are generated by misediting due to specific events of misguiding. Misediting can occur through the interaction of inappropriate gRNAs with mRNAs or appropriate gRNAs in an incorrect fashion. Four possible mechanisms for the generation of misedited sequences are presented. Chimeric molecules have been detected in steady-state mitochondrial RNAs that are composed of misguiding gRNAs covalently linked to mRNAs at misediting sites by the 3' oligo(U) tail. We propose that misediting within junction regions can be corrected by appropriately acting gRNAs.

Animals↗
PubMed1992

Computer methods for locating kinetoplastid cryptogenes.

RNA editing in the mitochondria of kinetoplastid protoza involves the insertion and/or deletion of precise numbers of uridine residues at precise locations in the numbers of uridine residues at precise locations in the transcribed RNA of certain genes. These genes are known as cryptogenes. In this paper we study computational algorithms to search for unknown cryptogenes and for the associated templates for insertion of uridines, gRNA sequences. The pairwise similarity search algorithm of Smith and Waterman (1) is modified to study this problem. The algorithm searches for unknown gRNAs given the cryptogene sequence. The method is tested on 4 known cryptogenes from L.tarentolae which are known to have 7 associated gRNAs. The statistical distribution of the longest gRNA when comparing random sequences is derived. Finally we develop an algorithm to search for cryptogenes using amino acid sequences from related proteins.

Algorithms↗
PubMed1992

Characterization of the [3H]-desipramine binding site of the bovine adrenomedullary plasma membrane.

The specific (i.e. nisoxetine-sensitive) binding of [3H]desipramine was studied in membranes prepared from bovine adrenal medullae. (1) [3H]desipramine bound reversibly and with high affinity (KD = 2.8 nmol/l) to a single class of non-interacting binding sites (Hill coefficient = 0.96); the maximal number of binding sites (Bmax) was 2.1 pmol/mg protein. (2) Binding of [3H]desipramine was dependent on [Na+] and [Cl-]. Increasing the concentrations of these ions increased binding. (3) Substrates and inhibitors of the neuronal noradrenaline transport system (uptake1) inhibited binding of [3H]desipramine with a rank order of potency typical for an interaction with the uptake1 carrier. The characteristics of [3H]desipramine binding remained essentially unchanged after solubilization of adrenomedullary membranes with the non-ionic detergent digitonin. The results indicate that the plasma membrane of bovine adreno-medullary cells is endowed with the neuronal uptake1 transporter.

Adrenal Medulla↗
PubMed1992

Antibody production in silkworm cells and silkworm larvae infected with a dual recombinant Bombyx mori nuclear polyhedrosis virus.

We have examined the efficiency of coexpression of two heterologous genes from a recombinant Bombyx mori nuclear polyhedrosis virus for the production of antibodies in silkworm larvae. The cDNAs encoding the light and the heavy chains of a murine immunoglobulin, directed against lipoprotein I of Pseudomonas aeruginosa, were brought under the control of two separate copies of the viral polyhedrin promotor. Infection of silkworm cells with the recombinant baculovirus yielded a maximum of 6.4 micrograms/ml IgG2A in the culture supernatant 72 hours post infection, while 800 micrograms/ml IgG2A was found in the hemolymph of infected fifth instar silkworm larvae seven days after infection with the same construct. The recombinant antibody exhibited a similar antigen specificity and avidity to that of the monoclonal antibody derived from ascites fluid.

Animals↗
PubMed1992

Facilitation of pattern recognition by cuing foveation with the luminance centroid as origin of the frame of reference.

The strategy for visual information processing must vary with the specific situation. We assume that in recognition of pre-learnt letter-like patterns under time-pressure conditions, mechanisms of selective attention are involved. We propose that, with simple stimuli, foveation is to the luminance centroids of such patterns, and if normally the latter has to be computed by the brain, cuing such information should improve performance. This assumption was tested on three subjects with five stimuli. In confirmation of the working hypothesis, we found significant improvements in performance (P = 0.01 or better) for each of the stimuli, with cuing to 'relevant' as compared with 'irrelevant' sites, and also when the point of foveation was nearer to the 'nodal' site. It is concluded that in pattern recognition processes, nodal regions are computed, the luminance centroid for example, for simple pattern stimuli.

Fovea Centralis↗
PubMed1992

Visual responses to reward-related cues in inferior parietal lobule.

A sample of 263 neurones was recorded in area 7a of the parietal lobe, in a monkey performing a reach task to visual targets displayed on a touch-sensitive videomonitor. The task had been operantly conditioned on food or juice rewards, and 78 (30%) of the units showed activity changes linked in some way to the reward. For most of these cells, the response was to the approach of the trainer's hand with the food reward. This specific visual response was similar irrespective of the direction of approach. Six cells increased discharge as soon as the task was completed in apparent anticipation of the reward. Another two neurones responded to missing a reward: they fired vigorously if the videoscreen was blanked in mid-trial because a target was not correctly touched. In many cases (40/78) the same cells responding to some aspect of the reward also responded to visual cues given during the task, especially the presentation of the target location. Reward-related activity in area 7a probably results from an integration of the visual and limbic inputs to this region, such that visual information which foretells behaviourally important events is emphasized.

Animals↗
PubMed1992

Hypothalamic etiology in sympathetically induced pathogenic cardiovascular changes in the cat.

Transient cardiovascular changes, including angina-like electrographic signs, are inducible by electrical stimulation of discrete sites in the prefornical region of the lateral hypothalamus (LH) of cats. Repeated stimulation may result in neurogenic cardiac infarction and myocardial bleeding. Thirty Swiss breed locally grown cats were studied under alpha-chloralose anesthesia in pairs, one as control and one experimental, with only the latter receiving multiple stimulations. ECG in lead II and arterial blood pressure were recorded on a Grass polygraph (USA). Electrical stimulation was delivered to a LH target at Fr 9.0, L2.5, D-1 to -2, by means of stainless steel wire electrodes. Stimulation consisted of 15-sec trains of square wave pulses at 100 Hz, 0.2-0.5 msec duration, and threshold currents of 0.05-0.1 mAmp. Stimulation at these LH sites induced small blood pressure changes, often with a small increase or no change in heart rate, presumably a manifestation of baroreceptor dysfunction. Repeating this stimulation greater than 6 times was shown to be pathogenic: on gross examination a darkened area was seen, mostly on the upper ventricular surface of the heart. Microscopic examination of such sites revealed subendocardial bleeding and sometimes also microinfarcts. Scanning electron microscopy revealed an unusually large number of contractures of the myofilaments. Biochemical analysis showed diffusion of catecholamines from nerves. Total myocardial blood flow increased following such stimulation, proportional to the stimulus intensity. It is pointed out that this is not contradictory to the sharply localized ischemic changes assumed to be responsible for the cardiopathy.

Adrenergic Fibers↗
PubMed1991

Chimeric gRNA-mRNA molecules with oligo(U) tails covalently linked at sites of RNA editing suggest that U addition occurs by transesterification.

Chimeric RNA molecules were detected by polymerase chain reaction amplification of kinetoplast RNA using a 3' primer specific to mRNA and a 5' primer specific to guide RNA (gRNA), and directly by Northern analysis. Covalent linkage of the 3' oligo(U) tail of the gRNA to the mRNA occurs at editing sites. Chimeric molecules were isolated for NADH dehydrogenase subunit 7 and cytochrome oxidase subunits II and III. We propose that these molecules are intermediates in the editing process and that successive transesterifications result in the transfer of uridine residues from the gRNA 3' oligo(U) tail to an editing site, with the number of uridine residues determined by base pairing with adenine and guanine "guide" nucleotides in the gRNA.

Animals↗
PubMed1991

The switch from larval to adult globin gene expression in Xenopus laevis is mediated by erythroid cells from distinct compartments.

The transition of hemoglobins during metamorphosis of Xenopus laevis involves replacement of the larval erythrocytes by adult ones, suggesting that the developmental control of this event depends upon the growth characteristics of the precursor cells. To identify the erythroid precursor cells and to investigate their developmental fate, we analyzed the distribution of stage-specific globin mRNAs by northern blotting in dorsal and ventral fragments of stage 32 embryos after in vitro culture as well as presumptive erythropoietic tissues of tadpoles during metamorphosis. The histological analysis shows that erythrocytes differentiate only in ventral fragments, suggesting that the ventral blood islands and most likely also the dorsolateral mesoderm are the primary sites of erythropoiesis. We also demonstrate that the first generations of erythrocytes, already express the predominating larval-specific alpha-globin mRNAs. The globin mRNA patterns obtained from presumptive erythropoietic tissues suggest an important role of circulating precursor cells in larval erythropoiesis, whereas the liver appears to be the main site of formation and maturation of the adult erythrocytes. Tentatively we propose that anuran erythropoiesis is dependent upon a self-perpetuating stem-cell line and that the larval and the adult erythrocytes are derived from successive generations of erythroid precursors, whose commitment may be imposed by the erythropoietic sites.

Animals↗
PubMed1990

Guide RNAs in kinetoplastid mitochondria have a nonencoded 3' oligo(U) tail involved in recognition of the preedited region.

Maxicircle-encoded guide RNAs (gRNAs) for cytochrome b and maxicircle unidentified reading frames 2 and 3 (MURF2 and MURF3) were isolated by hybrid selection and sequenced. All three gRNAs contained nonencoded 3' oligo(U) tails 5-24 nucleotides in length, with a mean length of approximately 15 nucleotides. Secondary structure calculations indicate a functional role of the 3' oligo(U) tail in stabilizing the initial hybrid formed between the gRNA and the preedited mRNA, and allowed the identification of potential mRNA recognition sites for an editing complex. In addition, isolated MURF2 gRNA-II could be 5' capped with [alpha-32P]-GTP and guanylyltransferase, suggesting that at least some gRNAs represent primary transcripts.

Animals↗
PubMed1990

A model for RNA editing in kinetoplastid mitochondria: "guide" RNA molecules transcribed from maxicircle DNA provide the edited information.

A class of small RNA molecules possibly involved in RNA editing is present in the mitochondrion of Leishmania tarentolae. These "guide" RNA (gRNA) molecules are encoded in intergenic regions of the mitochondrial maxicircle DNA and contain sequences that represent precise complementary versions of the mature mRNAs within the edited regions. In addition, the 5' portions of several gRNAs can form hybrids with mRNAs just 3' of the preedited region. A model is presented in which a partial hybrid formed between the gRNA and preedited mRNA is substrate for multiple cycles of cleavage, addition or deletion of uridylates, and religation, eventually resulting in a complete hybrid between the gRNA and the mature edited mRNA.

Animals↗
PubMed1990

Biochemical characterization and purification of the neuronal sodium-dependent noradrenaline transporter.

The protein properties of the neuronal sodium-dependent noradrenaline (NA) transporter of PC12 (rat pheochromocytoma) cells and of bovine adreno-medullary cells were studied by means of binding of 3H-desipramine (3H-DMI). 3H-DMI binding was decreased by proteases, phospholipase A2, by disulfide reducing agents and by the sulfhydryl-group alkylating agent N-ethylmaleimide. The NA transporter was partially purified by anion exchange and affinity chromatography. Tritiated desmethylxylamine (3H-DMX) bound irreversibly and in a DMI-sensitive manner to two PC12 membrane proteins (32kd and 53kd) which may represent components of the NA transporter.

Adrenal Gland Neoplasms↗
PubMed1990

Survival of pig embryos flushed from the reproductive tract immediately or two hours after slaughter of donors.

Four- to eight-cell embryos were collected from genital tracts of slaughtered gilts. Prepuberal donors were synchronized and superovulated by PMSG and hCG, inseminated and eventually slaughtered approximately 78 h after the expected time of ovulation. Genital tracts were flushed either immediately after slaughter (group A) or after 2 h storage at either 37 degrees C (group B) or 20 degrees C). Best results were achieved in group A, followed by group B and, by a wide margin, group C. Differences in viability of embryos became more evident with longer culture times.

Animals↗
PubMed1990

Reaching in the dark: enhanced responses in posterior parietal cortex.

In order to assess the relative importance of visual input to area 7 reach-related neuronal activity, a monkey was trained to reach to visual targets displayed on a video-monitor, both with and without visual feedback. Visual feedback was removed by having the monkey reach in darkness to a previously illuminated target. Of 19 reach-related cells recorded in area 7 both in the light and the dark, ten showed an enhancement of discharge in the dark. These included area 7b cells sensitive to screen contact and area 7a cells active during reach. Dark enhancement of active somatic responsiveness may partially compensate for the loss of visual guidance.

Animals↗
PubMed1989

Depressor effects of muscarinic and non-muscarinic mediation induced by lateral hypothalamic stimulation in the cat.

Transient sympathetically-mediated depressor effects were induced by stimulation of a small locus in the lateral hypothalamic peri-fornical region, medial to the fields of Forel. The ganglionic blocking agent, atropine methyl nitrate (ATMN), was used to show that muscarinic as well as non-muscarinic sympathetic ganglion receptor neurotransmission was involved. Evidence is presented that stimulation of this LH site co-activates a number of mechanisms and that depending on which of these are activated, the ganglionic blocking agent ATMN may either block, reverse or potentiate the depressor effect.

Animals↗