Biomedical subjects
Publications and source records attributed to C J Howe.
1. Procedures for multiple alignment of sequence data, subsequent phylogenetic inference, and testing of the trees derived are presented. 2. The assumptions underlying different approaches and the extent to which they are valid are discussed.
Cytoplasmic and thylakoid membranes have been purified from the cyanobacteria Anacystis nidulans R2 and Phormidium laminosum by sucrose density gradient centrifugation. Probing of Western blots of proteins from these purified membrane fractions with antibodies directed against the 33 kDa polypeptide of Photosystem II from pea indicates that this protein is present in both the thylakoid and cytoplasmic membranes, rather than just the thylakoid membranes. This has been confirmed by immunogold labelling of cells. Oxygen evolution assays have been used to show that the 33 kDa polypeptide is not assembled into a functional Photosystem II complex in the cytoplasmic membranes. This may be due to the absence of other Photosystem II components.
This study was undertaken to compare various extraction methods for insulin-like growth factor-binding proteins (IGFBPs) from insulin-like growth factor-I (IGF-I) in rat serum systematically, before measurement of IGF-I by radioimmunoassay (RIA). The values obtained in the IGF-I RIA following acid-ethanol (AE), acid-ethanol cryoprecipitation (AEC) and formic acid-acetone (FA) extraction methods were compared with the IGF-I values obtained following high-performance liquid chromatography (HPLC), which was the reference method. Radioligand blots were used to determine the pattern and degree of IGFBP removal by these methods. Over a wide range of circulating IGF-I levels, AE and AEC extraction gave IGF-I levels comparable with those obtained following HPLC. FA extraction resulted in IGF-I levels that were consistently higher (P < 0.01) than those obtained following HPLC and gave non-parallel displacement curves in comparison with recombinant IGF-I standards (P <0.01). Ligand blots demonstrated a similar pattern of IGFBP removal among the three methods with almost complete removal of IGFBP-3 but only 30-40% removal of the lower molecular weight IGFBPs. These lower molecular weight IGFBPs did not interfere with the RIA measurements of IGF-I from AE and AEC extracts. Therefore the AE extraction method of Daughaday, originally validated for use in human serum, is also satisfactory for use in rat serum. The complete removal of IGF-binding activity does not appear essential for accurate measurement of IGF-I by RIA, although this may depend on the specific binding characteristics of the IGF-I antiserum.
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The assessment and management of a child's pain in the postoperative period remain a challenge to the PACU nurse. Assessment of pain in children is often difficult; ongoing clinical research in this area continues to expand knowledge of how to improve children's communication of their pain to caregivers. Many pain management strategies are available, both pharmacologic and nonpharmacologic, to ease pain and distress postoperatively. Through understanding and knowledge of pain, its assessment and its management, the PACU nurse contributes positively to the child's surgical experience.
This article is a report of our experience with an interdisciplinary pain service in a large tertiary care pediatric hospital. During the first 2 years of operation, we received 869 consultations and referrals from more than 19 hospital divisions. Postoperative pain was the most frequent reason for consultation (56% of patients). Patients with pain related to cancer and sickle cell disease comprised 25% of the consultations. The remaining patients had a wide variety of primary diagnoses and causes of pain. We calculated the time spent by pain service physicians in direct patient care. The majority (63%) of physician time was spent with a small number of patients (17%). Most of these patients had pain that was unrelated to surgery, cancer, or sickle cell disease, and many posed dilemmas in diagnosis and treatment. Physician time was correlated directly to the use of psychologic and physical therapies for the pain, involving multiple team members. This experience supports the demand for an interdisciplinary pain service in a tertiary care children's hospital. A significant amount of physician time is necessary to provide patient care and to maintain a team approach, however, and pediatricians and other health care professionals who aim to implement such services should be cognizant of the time required.
The thylakoid processing peptidase from the cyanobacterium Phormidium laminosum has been extracted from thylakoid membranes by solubilization with Triton X-100. Its reaction specificity has been compared with the analogous pea peptidase by processing in vitro of radiolabelled wheat and P. laminosum thylakoid lumenal precursor polypeptides. The cyanobacterial polypeptide is processed to the mature size through an intermediate by the P. laminosum peptidase, but to a polypeptide that has a slightly greater apparent molecular weight than the intermediate by the pea peptidase. Both peptidases correctly process the wheat polypeptide. This suggests that the reaction specificities of the two peptidases are similar, but not identical.
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A 9 kDa polypeptide which is loosely attached to the inner surface of the thylakoid membrane and is important for the oxygen-evolving activity of Photosystem II in the thermophilic cyanobacterium Phormidium laminosum has been purified, a partial amino acid sequence obtained and its gene cloned and sequenced. The derived amino acid sequence indicates that the 9 kDa polypeptide is initially synthesised with an N-terminal leader sequence of 44 amino acids to direct it across the thylakoid membrane. The leader sequence consists of a positively charged N-terminal region, a long hydrophobic region and a typical cleavage site. These features have analogous counterparts in the "thylakoid-transfer domain" of lumenal polypeptides from chloroplasts of higher plants. These findings support the view of the proposed function of this domain in the two-stage processing model for import of lumenal, nuclear-encoded polypeptides. In addition, there is striking primary sequence homology between the leader sequences of the 9 kDa polypeptide and those of alkaline phosphatase (from the periplasmic space of Escherichia coli) and, particularly in the region of the cleavage site, the 16 kDa polypeptide of the oxygen-evolving apparatus in the thylakoid lumen of spinach chloroplasts.
Following exposure to UV light the synthesis of six polypeptides (112, 100, 89, 76, 71 and 65 kDa) was found to be enhanced or induced in the alga Chlamydomonas reinhardtii. Treatment with 4-nitroquinoline-N-oxide (NQO) resulted in the enhanced/induced synthesis of six polypeptides with molecular masses similar to those enhanced following exposure to UV light. Heat shock resulted in the enhanced synthesis of five polypeptides (89, 76, 71, 60 and 22 kDa), three of which (89, 76 and 71 kDa) had apparently identical mobilities to polypeptides whose synthesis was enhanced following UV treatment.
Previous research has suggested that visual information has primacy over verbal and vocal information in the judgement of social attitude. However, this could be because the contextual inappropriateness of the verbal information switched attention away from the auditory channel and on to the visual. The experiment reported in this paper assesses this possibility by considering the contribution made by visual information when the verbal is appropriate. Focusing on the expression of superior and inferior attitude, it finds that visual information does not simply fail to achieve primacy. It is markedly less important than either vocal or verbal information, which both play more or less equal roles.
We have determined the DNA sequences of regions involved in two of the three inversions known to have occurred during the evolution of wheat chloroplast DNA. This establishes the extent of the second largest of the three inversions. Examination of these sequences suggests that although short repeated sequences are present, the endpoints of the second and third inversions are not associated with repeated sequences as long as those associated with the first inversion. However the endpoints of all three inversions are all adjacent to at least one tRNA gene, and there is evidence that three of the tRNA genes have been subjected to partial duplication, possibly at the time of inversion. This suggests that tRNA genes might be involved with rearrangements of chloroplast DNA, as has also been postulated for mitochondrial DNA.
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The major purpose of this investigation was to determine the effect of prednisolone and megestrol acetate in cats on the adrenal cortisol response to exogenous adrenocorticotrophic hormone during drug administration at dose rates employed for management of some inflammatory feline dermatoses. Prednisolone (at least 2 mg/kg/day) and megestrol acetate (5 mg/cat/day) were each administered orally to seven cats from days 1 to 16. Three additional cats received no therapy. Basal and stimulated cortisol concentrations, food and water intake, hematology, blood biochemistry, urinalyses, and hepatic and cutaneous histology were studied in all cats before, during, and two weeks following the end of treatment. Cats given prednisolone or megestrol acetate had significant suppression of stimulated cortisol levels on day 8. This change was more marked on day 15, when the suppression in cats given megestrol acetate was also significantly more severe than in those receiving prednisolone. Recovery of adrenal reserve was considered present on day 30 in six of seven cats given prednisolone, but in only three of seven receiving megestrol acetate. Eosinopenia, glycosuria and hepatocyte swelling from glycogen deposition were occasionally recorded in treated cats of both groups, providing additional circumstantial evidence for glucocorticoid activity of megestrol acetate in cats. It is advised that abrupt withdrawal of prednisolone or megestrol acetate therapy be avoided in this species to reduce the chance of precipitating clinical signs of hypoadrenocorticism, even after treatment for as little as one week.
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The endpoints of an inversion in wheat chloroplast DNA are shown to be associated with copies of a short repeated sequence. Recombination across the repeats in an inverted configuration may have been responsible for the inversion, although they are currently in a direct orientation owing to a second inversion. The repeated sequence contains an element homologous to the core of the bacteriophage lambda att-site, which can function as such in vivo.
The mitochondrial genome of maize contains a DNA sequence homologous to the chloroplast gene coding for the large subunit of ribulose-1,5-bisphosphate carboxylase (LS gene). The presence in mitochondrial DNA of both coding and flanking sequences of this gene has been demonstrated first, by cross hybridization between the purified organelle DNAs and between cloned mitochondrial and chloroplast DNA sequences and second, by in vitro transcription-translation of cloned mitochondrial DNA in an E. coli cell free system where a 21,000 dalton polypeptide is synthesized that can be precipitated with antibodies to wheat ribulose-1,5-bisphosphate carboxylase. In contrast to the 12 kb chloroplast homologous sequence found in the mitochondrial genome (Stern and Lonsdale, 1982), the sequence homologous to the LS gene is unaltered in mitochondrial DNA isolated from the male sterile cytoplasms of maize. The LS gene homologous sequence in the mitochondrial genome is located some 65 kb from the 18S mitochondrial rRNA gene and approximately 20 kb from the mitochondrial DNA sequence having homology to the chloroplast 16S rRNA gene.
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