Biomedical subjects
Publications and source records attributed to J Haley.
These electrophysiological studies, using desensitization and selective antagonists, demonstrate a peripheral role of bradykinin in the generation of pain in the rat. In addition, nitric oxide is shown to play a complex role in peripheral and spinal events in nociception.
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The activity of single dorsal horn nociceptive neurones was recorded in the spinal cord of halothane anaesthetized rats. S.c. injection of a 5% formalin solution into the receptive field of these neurones resulted in two peaks of neuronal firing over a period of 60 min. Prior administration of the kappa-opioid receptor agonist U50488H directly into the site of formalin injection caused a dose-dependent decrease in the size of both the first and second peaks of the response which was naloxone reversible. Injection of U50488H into the contralateral paw had no effect on either peak of the formalin response. Injection of the top dose (100 micrograms) of U50488H had no effect on the electrically evoked A beta- or C-fibre responses of the neurone. Neither morphine nor Tyr-D-Ser(Otbu)-Gly-Phe-Leu-Thr (DSTBULET), administered into the receptive field, had any significant effect on either peak of the formalin response. Plasma extravasation in the skin, measured using Evans blue, produced by the formalin injection was not blocked by U50488H. Thus, whilst the spinal responses of this peripheral model of inflammation can be inhibited by peripheral kappa-opioid activation, but not mu- or delta-, plasma extravasation associated with this inflammation is not reduced.
The epidermal-growth-factor (EGF) receptor was expressed in the human erythroleukaemic cell line K562 by transfection of the receptor cDNA. EGF-receptor biochemistry appears altered in the K562 transfectants. Autophosphorylation of the K562 receptor is not stimulated substantially by EGF. Tyrosine kinase activity of the receptor is high in the absence of EGF, whereas receptor affinity for EGF is low. K562 cells are shown to lack mRNA for transforming growth factor alpha (TGF alpha). Therefore autocrine stimulation of the K562 receptor, at least by TGF alpha, does not explain the observed receptor biochemistry. The K562 receptor is phosphorylated at a single major site in intact cells, a threonine residue that may be Thr-669. Possible mechanisms of regulation of the EGF receptor in the K562 transfectants are discussed.
Many chloroplast genes are co-transcribed in polycistronic transcription units that give rise to numerous overlapping RNAs, but the significance of this pattern of transcript accumulation is not understood. An analysis of the transcripts of the adjacent and divergent maize psbE-psbF-psbL-ORF40 and ORF31-petE-ORF42 gene clusters indicates that transcription initiation at alternative promoters contributes to the generation of overlapping RNAs for both clusters. Furthermore, developmentally varying transcript ratios for the ORF31-petE-ORF42 gene cluster are determined at least in part by selective promoter usage. During light-induced plastid maturation, increased levels of primarily monocistronic petE transcripts accumulate from a promoter upstream of the internal petE gene. Dark-predominant and non-light-responsive bi- and tricistronic transcripts result from transcription initiation upstream of ORF31, the proximal gene of the cluster. In addition to the transcriptional overlap within gene clusters, divergent transcription units for the two gene clusters overlap and reciprocal antisense RNAs accumulate. The organization of the transcription units in this region raises the possibility of promoter interdependence or other functional interaction between transcription units.
We investigated the following three assumptions regarding anxiety in hospitalized children: (1) anxiety decreases during hospitalization, (2) anxiety correlates with symptoms, procedures, and parental anxiety, and (3) anxiety is reduced following guided play with real and simulated medical equipment. Fifty latency-age children and their parents were studied. Anxiety was measured by self-report, parental report, nurse's report, and direct observation. Potential correlates were monitored daily by review of patient care records, and interviews of primary nurses and parents. Hypothesis one was confirmed; anxiety decreased significantly (p less than 0.001) across the four assessments. Hypothesis two was confirmed; symptoms, procedures, and parental anxiety accounted for 27-30% of the variance in children's anxiety. Hypothesis three was not confirmed; anxiety decreased following guided play, but not enough to reach statistical significance.
A 66 kd protein, pl 5.4, was purified from the Triton-insoluble fraction of rat spinal cord. This protein formed 10 nm filaments in vitro. The 66 kd protein was unique, although it shared homology with the 70 kd neurofilament protein (NF-L) and vimentin. An antiserum (anti-66) specific to the 66 kd protein did not cross-react with any of the neurofilament triplet proteins. In the spinal cord, anti-66 intensely stained the axons of the anterior and lateral columns. However, afferents from dorsal root ganglia and the efferents from the motoneurons were negative. In the cerebellum, anti-66 intensely stained most axons. The 66 kd protein was readily detectable in homogenates of forebrain, cerebellum, brainstem, and spinal cord, but was found only in trace amounts in adult sciatic nerves and was not found in extraneural tissues. The 66 kd protein constituted 0.5% of total protein in the spinal cord, whereas NF-L constituted about 1.5%.
Four polypeptides, three of which are chloroplast-encoded, have been shown to be associated with the thylakoid membrane cytochrome b6-f complex. In this report, the gene for a fifth polypeptide, which copurifies with the b6-f complex, is identified through the use of an antibody generated against a synthetic decapeptide predicted from a maize chloroplast DNA sequence. The deduced 37-amino acid sequence of the immunoreactive 4-kDa polypeptide is 100% and 86% conserved in the respective similar open reading frames encoded by Nicotiana tabacum and Marchantia chloroplast DNA. The 4-kDa polypeptide is present in both etioplasts and chloroplasts of maize and is found as well in spinach, tobacco, pea, wheat, and rice thylakoids. Similar to the other subunits of the b6-f complex, it is intrinsic to the membrane, and its hydrophilic COOH terminus is located at the stromal thylakoid surface. We propose to call the 4-kDa polypeptide "subunit 5" and the chloroplast gene that encodes it the petE gene.
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A porcine genomic cosmid library was constructed to study the structure and regulation of the porcine relaxin gene. Two overlapping cosmids containing relaxin-specific sequences were isolated, and a 9-kilobase BamHI fragment containing the porcine relaxin gene was sequenced. The sequence derived from these cosmids was used to characterize the relaxin gene transcription unit utilized in the pregnant ovary. Analysis of chromosomal and cosmid DNA has shown that porcine relaxin is encoded by a single copy gene, comprising two exons separated by a 5.5-kilobase intron. The gene contains elements representing two classes of repeated sequences homologous with the human Alu and Kpn families. The transcriptional initiation site of this gene was located by primed extension and direct sequencing of relaxin-specific cDNA. Furthermore, the promoter elements were mapped and sequenced, and transcriptional activity was demonstrated. The promoter contained sequences with homology to regions conserved within retroviral long terminal repeats and within the adenovirus E4 promoter.
Epidermal growth factor (EGF) receptor expression in 31 primary human bladder tumours was quantitated using both structural and functional assays and the EGF receptor gene in the same tumours was analyzed by Southern blot analysis. Immunocytochemical studies using the EGFR1 monoclonal antibody (Mab) showed a significant correlation between EGF receptor levels and the stage and grade of the tumours. Autophosphorylation assays employed to evaluate the receptor's tyrosine kinase activity gave results which in general were consistent with the immunocytochemical data. Using internally controlled immunocytochemical studies with two Mabs and Southern blot analysis of DNA isolated from the tumours, no evidence was obtained for the production of truncated receptors similar to those encoded by the v-erb-B oncogene. Gene amplification was not found in any of the superficial tumours, but one invasive tumour with high EGF receptor expression had an 8-10 fold amplification of the EGF receptor gene. The EGF receptor isolated from this tumour showed a normal pattern of tyrosine phosphorylation at all three major autophosphorylation sites. Our detailed study is consistent with the correlation previously found between EGF receptor expression and stage and grade of bladder tumours, and suggests that at this level of analysis EGF receptors in bladder tumours are not abnormal in structure or size, autophosphorylation activity, or gene structure.
Relaxin is a peptide hormone produced by the corpora lutea of ovaries during pregnancy, softening and lengthening the ligaments of the pelvis and softening the cervix in order to make childbirth easier. In attempts to determine the nucleotide sequence coding for relaxin, recombinant DNA techniques were used to obtain a cDNA clone bank from total mRNA isolated from the ovaries of pigs in late pregnancy. Clones were screened using cDNA initiated by synthetic oligonucleotide primers coding for the Trp Val Glu Ile sequence of the porcine relaxin B chain. The synthetic undecamer [5'-ATCTCCACCCA-3'] was found to prime a specific 32P-labeled cDNA of approximately 300 nucleotides containing B chain and signal peptide coding sequences, as verified by nucleic acid sequence analysis. This cDNA was used to probe the ovarian clone bank. Several clones containing large inserts which hybridized to this probe were subjected to sequence analysis and some of these were found to contain the preprorelaxin coding region, comprising a signal peptide of 24 amino acids, a B chain of 32 amino acids, a large C peptide of 104 amino acids, and an A chain of 22 amino acids. From the amino acid sequence of prorelaxin derived in this way, it appears that the processing of prorelaxin involves two enzymes with chymotrypsin-like and trypsin-like specificity, respectively. In comparisons of porcine and rat preprorelaxins, the C region had as much amino acid sequence homology as the B and A chains. The C region is also rich in charged amino acids, suggesting a role for it beyond simply ensuring proper disulfide bond formation.
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Using a synthetic oligonucleotide primer, cloned DNA fragments, each containing the entire coding sequence of rat relaxin, have been isolated from a clone 'bank' of ovarian mRNA sequences. The nucleotide sequence of these clones demonstrates that relaxin is synthesized as a preprorelaxin molecule with an unexpectedly large connecting peptide of 105 amino acid residues.
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This article is a description of different approaches to therapy with a family orientation. There are general categories of family therpay which had their origins in individual therapy, such as the approaches based upon psychodynamic theory, those derived from experiential procedures, and the behavioral approaches. There are also family therapies which have not developed from individual therapy, such as the extended family system approach and the communication school of family therapy. The different therapy approaches are described within a set of dimensions which characterize most therapy. Such dimensions include whether the past or present is emphasized, whether the therapist uses interpretation or directives, whether the approach is in terms of growth or specific problems, whether hierarchy is a concern, and whether the unit is an individual, two people, three people, or a wider network. Illustrations of the different family therapy approaches are given in terms of the kinds of information that would interest the therapist of each school and the kinds of actions he or she would take to bring about change.
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