Biomedical subjects
Publications and source records attributed to S Roberts.
The complete sequence of the HindIII-BamHI/BglII fragment of plasmid pSV2 gpt, which contains the E. coli gpt gene, has been determined and used to predict the amino acid sequence of the protein. The nucleotide sequence was used to create a map of eight gpt gene deletion mutants which were generated in mammalian cells.
Scripts of a short answer Final MB examination paper were re-marked by the original examiner and three colleagues. While individual question marks varied considerably, most examiners agreed quite well as to the overall rank order of candidates--although not about absolute total marks--but a few disagreed considerably. Examiners exhibited remarkably consistent idiosyncratic behaviour in terms of the distribution of marks awarded. These factors led to haphazard pass/fail and pass/distinction decisions on this particular component of the examination. Recommendations are put forward to improve the marking consistency and use of short answer question papers.
This paper makes three main points: There are many similarities between man-made rules for detecting causality and "laws" of classical conditioning, presumably generated by nature. They suggest that the function of classical conditioning resembles the detection of causality. There are twelve similarities between the clock used by rats to discriminate time and a stopwatch. They suggest that the rat's clock, like a stopwatch, was designed for measuring unknown durations. There are eight similarities between time discrimination and classical conditions. In particular, whether or not a stimulus is timed depends on the signal value of the stimulus. The similarities suggest that time discrimination and classical conditioning - their underlying mechanisms - have similar functions.
Uptake of D-[14C]glucose and D-[14C]xylose by Spirochaeta aurantia was demonstrated to be osmotic shock sensitive and to require a high-energy phosphorylated compound rather than a proton motive force. These features are similar to those of binding protein-mediated transport systems in other gram-negative bacteria.
Forty-two patients with frozen shoulder were followed up closely for eight months. They were all taught pendular exercises and randomly allocated to one of four treatment groups: (a) intraarticular steroids, (b) mobilisations , (c) ice therapy, (d) no treatment. This study has shown that there is little long-term advantage in any of the treatment regimens but that steroid injections may benefit pain and range of movement in the early stages of the condition.
As the natural history of frozen shoulder is poorly documented, a prospective study of 40 patients followed up for 40-48 months (mean 44 months) is described. The range of movement was significantly less than age- and sex-matched controls. Objective restriction was severe in five patients and mild in a further 11. Patients were often unaware that shoulder range was impaired. Dominant arm involvement, manual labour, and mobilisation physiotherapy were associated with a less satisfactory outcome. We conclude that, while objective restriction persists, there is little functional impairment in the late stage of frozen shoulder.
Two distantly related, two- to three-year-old male Akita dogs developed partial or complete night blindness which progressed to day blindness. Clinical features were attenuated retinal blood vessels, diffuse exaggerated tapetal reflex, and hyperreflective horizontal lines in the tapetal fundus. The principal histologic change was regional photoreceptor cell degeneration. Areas of moderately atrophic sensory retina alternated, concentrically to the papilla, with areas of severely atrophic retina. Ultrastructurally, there was disorganization of photoreceptor outer segments, retinal gliosis, focal loss with reorganization of the outer limiting membrane, and retinal invasion by pigment epithelium-derived macrophages. Clinical observations on five additional Akitas with generalized retinal atrophy suggest that the condition is heritable.
Five experiments with rats investigated under what conditions a stimulus is timed by the internal clock used in time-discrimination procedures. In Experiments 1-4, we trained rats to time one stimulus (e.g., light) and then asked whether they timed a stimulus from another modality (e.g., sound). The second stimulus was treated in three ways: exposed (presented alone), paired with food, and extinguished. Experiments 1 and 2 used the peak procedure, similar to a discrete-trial fixed-interval schedule, and paired the treated stimulus with food using instrumental training; Experiments 3 and 4 used a psychophysical choice procedure and paired the treated stimulus with food using classical conditioning. All four experiments found that there was cross-modal transfer of the time discrimination after pairing, but not after exposure or extinction. This suggests that the rat's internal clock timed the treated stimulus after pairing, but not after exposure or extinction. Experiment 5 tested a theory of extinction based on the results of Experiments 1-4; the results suggested that the decline of responding observed in extinction is not due to changes in timing. The main conclusion is that the internal clock apparently times stimuli with signal value (associative strength) and does not time stimuli without signal value.
A porcine prorelaxin gene has been constructed partly by synthetic means and partly from its natural messenger RNA. A gene coding for the 32 N-terminal amino acids including a chain initiator methionine codon (B gene) was synthesised and inserted in a plasmid at a site downstream from a tryptophan promoter in such a way that its expression is under the control of the trp promoter. DNA corresponding to the rest of the prorelaxin was prepared using reverse transcriptase extension of a primer complementary to relaxin mRNA and joined at a suitable restriction site to the B gene. Transformation of E. coli with this plasmid followed by suitable induction resulted in the synthesis of a new protein identified as prorelaxin by its size and its antigenic similarity to relaxin.
The relation between preclinical tripos and clinical examination results and subsequent career success of 188 medical graduates of Cambridge University was measured using five indicators of success. A generally positive relation was found, but this was not specific enough to make accurate individual predictions. Present levels of appointment were more closely related to clinical than preclinical results. No support was found for the local assertion that "2.1s" do better than "firsts" in clinical medicine. Since undergraduate examination results seem to be inaccurate predictors of later performance they should not be used as the principal evidence in making selection decisions.
We have investigated the chromosomal location, inheritance, and expression of a cloned rabbit beta-globin gene introduced into the mouse germ line by microinjection into mouse eggs. Experiments utilizing in situ hybridization to metaphase chromosomes show that the gene has integrated into one or two different chromosomal loci in each of five mouse lines analyzed. Each locus contains between three and forty copies of the foreign DNA sequence arranged in a tandem array, and the sequences at each locus are stably inherited as a single Mendelian marker. Neither globin mRNA nor polypeptides encoded by the rabbit beta-globin gene are detected in erythroid cells in the seven transgenic lines examined, indicating that the expression of the foreign gene is not correctly regulated. However, in two of the mouse lines, rabbit beta-globin transcripts are found at a low level in specific, although inappropriate, tissues: skeletal muscle in one line and testis in another line. These unusual patterns of beta-globin gene transcription are heritable traits in the two mouse lines and may result from the beta-globin gene's integration at abnormal chromosomal positions.
Ribosomal protein phosphorylation was investigated in isolated ribosomal subunits and polyribosomes from rat cerebral cortex in the presence of [gamma-32P]ATP and purified catalytic subunit of cyclic AMP-dependent protein kinase from the same tissue. Ribosomal proteins that were most readily phosphorylated in isolated cerebral ribosomal subunits included proteins S2, S3a, S6 and S10 of the 40 S subunit and proteins L6, L13, L14, L19 and L29 of the 60 S subunit. These proteins were also phosphorylated in cellular preparations of rat cerebral cortex in situ or in vitro [Roberts & Ashby (1978) J. Biol. Chem. 253, 288-296; Roberts & Morelos (1979) Biochem. J. 184, 233-244]. However, several additional ribosomal proteins were phosphorylated when isolated 40 S or 60 S subunits were separately incubated in the reconstituted system. Analogous results were obtained with an equimolar mixture of cerebral 40 S and 60 S subunits under comparable conditions. In contrast, extensive exposure of purified cerebral polyribosomes to the catalytic subunit resulted in phosphorylation of only those ribosomal proteins of the 40 S subunit that were most highly labelled after the administration of [32P]Pi in vivo: proteins S2, S6 and S10. Ribosomal proteins of 60 S subunits that were readily phosphorylated in isolated cerebral polyribosomes included proteins L6, L13 and L29. These results indicate that polyribosome formation markedly decreases the number of ribosomal protein sites available for phosphorylation by the catalytic subunit of cyclic AMP-dependent protein kinase. Moreover, the findings suggest that, of the ribosomal protein phosphorylations observed in rat cerebral cortex in vivo, proteins S2, S6, S10, L6, L13 and L29 can be phosphorylated in polyribosomes, whereas proteins S3a, S5, L14 and L19 may become phosphorylated only in free ribosomal subunits.
Studies were carried out to elucidate the mechanisms underlying the diminished phosphorylation of cerebral ribosomal protein in experimental hyperphenylalaninaemia [Roberts & Morelos (1980) Biochem. J.190, 405-419]. Administration of N(6),O(2)'-dibutyryl cyclic AMP or 3-isobutyl-1-methylxanthine, which increased phosphorylation of the S6 protein of cerebral 40S ribosomal subunits in control infant rats, did not counteract the decreased phosphorylation of this ribosomal protein resulting from intraperitoneal administration of a loading dose of l-phenylalanine. N(2),O(2)'-Dibutyryl cyclic GMP had no effect on cerebral ribosomal-protein phosphorylation in either control or hyperphenylalaninaemic animals. The phenylalanine-induced decrease in ribosomal-protein phosphorylation was associated with decreased protein kinase activity in cerebral cytosolic and microsomal preparations. However, the maximal protein kinase response to cyclic AMP added in vitro was unaltered by prior administration of phenylalanine in vivo. The heat-stable protein inhibitor of cyclic AMP-dependent protein kinases decreased the activity of these enzymes by about 90% and eliminated the phenylalanine-induced difference in protein kinase activity in the absence of added cyclic AMP. Intracisternal administration of doses of dibutyryl cyclic AMP or 3-isobutyl-1-methylxanthine which increased the cyclic AMP-dependent protein kinase activity ratio in control infant rats was without effect on this index in phenylalanine-treated animals. Dibutyryl cyclic GMP had no effect on the protein kinase activity ratio in either group of animals. These results suggest that inhibition of cerebral cyclic AMP-dependent protein kinases by abnormally high concentrations of phenylalanine may contribute to the decrease in cerebral ribosomal-protein phosphorylation in experimental hyperphenylalaninaemia.
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The usefulness of small bowel enema X-ray examination was evaluated in 11 patients in whom a provisional diagnosis of incomplete small bowel obstruction was made. Dilute barium was introduced through a nasojejunal tube. In one patient intubation of the jejunum was unsuccessful. In six patients the suspected obstruction was confirmed and information obtained about the site of obstruction. The small bowel enema X-ray examination was normal in three patients and subsequent clinical outcome was uneventful. In one patient, with normal findings on small bowel enema X-ray examination, acute obstruction developed after two days, necessitating operation. Small bowel enema X-ray examination appears a worthwhile adjuvant to clinical assessment but decisions for operative intervention still need to be based on clinical evaluation of patients.
This study on the phosphorylation in vivo of membrane proteins in cerebral cortices of infant rats reports the identification of the adrenocorticotropin (ACTH)-sensitive phosphoprotein B-50 as one of the substrate proteins that are rapidly phosphorylated in vivo following intracisternal administration of 2 mCi [32P]orthophosphate. Rats were sacrificed 30 min after isotope injection. A fraction enriched in membranes, designated neural membranes (NM), was isolated from the cerebral cortices according to the procedure used for preparation of synaptic plasma membranes (SPM) from adult brain. This NM fraction was characterized by electron microscopy. The proteins of NM were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Numerous protein bands of NM in infant rat brain were phosphorylated in vivo. Attention was focussed on the 32P-labeled protein bands in the molecular weight range of 47K-67K. In this region one phosphoprotein band (MW 48K) was more highly labeled than the other bands. The electrophoretic behavior of three of these labeled bands, designated a, c, and e (MW 48K, 55K, and 62K, respectively) was compared with that of protein bands that were phosphorylated in vitro in cerebral membranes isolated from noninjected infant rats. The effects of ACTH1-24 and cyclic AMP in the in vitro system were also studied to probe for the presence of specific membrane proteins known to be sensitive to these modulators. On incubation of NM with [gamma-32P)ATP in the presence and absence of ACTH1-24 in vitro, phosphorylation of a 48K protein band was inhibited in a dose-dependent fashion by the neuropeptide. Two-dimensional electrophoretic separation of NM proteins labeled in vivo indicated that the 48K band had an isoelectric point of 4.5, identical to that of the ACTH-sensitive B-50 protein previously identified. Cyclic AMP stimulated phosphorylation in vitro of two protein bands (MW 55K and 59K) in NM preparations. This result indicates that the in vivo labeled band c may correspond to the cyclic AMP-sensitive 55K protein, whereas phosphoprotein band e, labeled in vivo, appears to be different from the cyclic AMP-sensitive 59K protein band. These observations indicate that neural membranes isolated from infant rat cerebral cortices contain a variety of proteins that can be phosphorylated in vivo. Several of these, for example, the 48K protein band, have the properties of synaptic plasma membrane proteins of adult rat brain that have been characterized by their sensitivity to neuromodulators in endogenous phosphorylating systems in vitro.
Surgically removed discs from patients with either spondylolisthesis or tears of the posterior annulus fibrosus were analysed for water, proteoglycan, and collagen content and compared with post-mortem control material. Discs from patients with spondylolisthesis had a reduced proteoglycan content in all section sampled and less collagen in the outer annular layers. In contrast discs containing tears in the posterior annulus were unaltered biochemically, although extended studies on 2 patients indicated that there may be localised biochemical changes in the region of the tear itself. Collagen types I, II, and III and proteoglycan distributions were studies qualitatively by immunofluorescence. Collagen types I and III appeared to be reduced in discs from patients with spondylolisthesis, but again little change was found in patients with tears in the posterior annulus fibrosus.
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