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Biomedical subjects

G M Thompson

Publications and source records attributed to G M Thompson.

At least 19 recordsLinked to original sources
PubMed1999

The beta subunit of eukaryotic translation initiation factor 2 binds mRNA through the lysine repeats and a region comprising the C2-C2 motif.

Eukaryotic translation initiation factor 2 (eIF2) has been implicated in the selection of the AUG codon as the start site for eukaryotic translation initiation, since mutations in its three subunits in yeast that allow the recognition of a UUG codon by the anticodon of the initiator Met-tRNAMet have been identified. All such mutations in the beta subunit of eIF2 (eIF2beta) mapped to a region containing a putative zinc finger structure of the C2-C2 type, indicating that these sequences could be involved in RNA recognition. Another feature of eIF2beta that could mediate an interaction with RNA is located in the amino-terminal sequences and is composed of three repeats of seven lysine residues which are highly conserved in other species. We show here the ability of eIF2beta, purified from Escherichia coli as a fusion to glutathione S-transferase, to bind mRNA in vitro. Through a deletion analysis, mRNA binding was found to be dependent on the lysine repeats and a region encompassing the C2-C2 motif. Strong mRNA binding in vitro could be maintained by the presence of only one lysine or one arginine run but not one alanine run. We further show that only one run of lysine residues is sufficient for the in vivo function of eIF2beta, probably through charge interaction, since its replacement by arginines did not impair cell viability, whereas substitution for alanines resulted in inviable cells. mRNA binding, but not GTP-dependent initiator Met-tRNAMet binding, by the eIF2 complex was determined to be dependent on the presence of the lysine runs of the beta subunit.

Animals↗
PubMed1998

Effect of exposure to balanced salt solution upon the hardness of the crystalline lens.

BACKGROUND: The nucleus confers most of the hardness upon the lens, water content decreases towards the centre of the nucleus and a relative dehydration accompanies increased hardness in some cataractous lenses. It is a possibility that exposure and incubation of the inner layers of the nucleus to balanced salt solution (BSS) can result in the softening of the nucleus. PURPOSE: This study aimed to investigate the effect of BSS upon the lens hardness. METHODS: Nuclear colour element of cataract was graded biomicroscopically. Following extracapsular cataract surgery the lens nucleocortex was divided into two equal parts and each half was allocated randomly to incubation in BSS or air for 5 min before the hardness of each section was assessed by an automated guillotine. RESULTS: Following incubation with BSS the mean force necessary to bisect the lens was 0.50 N and in the control air group the mean force was 0.64 N. The lenses in the BSS-treated group were consistently softer than those in the control group, with a mean softening of 18.3% (P=0.001). The amount of softening was not related to the nuclear colour (P=0.6) or age (P=0.1). CONCLUSION: Softening of the lens through physical disruption has previously been reported. This study describes the phenomenon of nuclear softening following exposure to BSS, indicating that lens softening can occur through biochemical means.

Aged↗
PubMed1998

Desensitization of beta3-adrenergic receptor-stimulated adenylyl cyclase activity and lipolysis in rats.

The beta3-adrenergic receptor is an integral membrane protein consisting of seven transmembrane domains. Unlike the beta1 and beta2 receptors, this subtype lacks the consensus phosphorylation sites required for desensitization by serine kinases. Using the rodent specific beta3 agonist BRL 35135, our initial data indicated that beta3 receptor-mediated glycerol levels progressively decreased following daily oral doses of 5 mg/kg. Therefore, we initiated studies designed to delineate the possible mechanism(s) for this decreased response. Within 3 hours following a single oral dose of BRL 35135, serum glycerol levels and UCP (uncoupling protein) RNA levels were significantly increased whereas beta3 RNA levels were significantly decreased. Rats were dosed daily for 5 days with either vehicle or BRL 35135 (5 mg/kg, p.o.) and blood samples were collected for glycerol analysis. Adipose tissue was excised for lipolysis and adenyl cyclase measurements. In addition, UCP and beta3 receptor RNA levels were assessed. No effect on adipocyte BRL 37344-stimulated adenylyl cyclase activity was observed 3 hours following the initial dose of BRL 35135. Although a slight decrease (approximately 25%) in adenylyl cyclase activity could be observed 24 hours following the initial dose, it wasn't until day 4 of dosing that a significant decrease (50%) was observed. In contrast, beta3- stimulated lipolysis in adipocytes from BRL 35135-treated rats was decreased 85% within 24 hours and this decrease persisted through four days of treatment. These data indicate that the lipolytic response to beta3 receptor activation is decreased after only a single oral dose of BRL 35135, whereas receptor-mediated adenylyl cyclase activation, although initially unaffected, also desensitizes by day four of treatment.

Adenylyl Cyclases↗
PubMed1998

Does postsynaptic alpha 1-adrenergic receptor supersensitivity contribute to autonomic dysreflexia?

Quadriplegics often experience periods of severe hypertension known as autonomic dysreflexia. Clinically, these events have been well documented, but the mechanisms for mediating autonomic dysreflexia remain unclear. We used a chronic rat model to investigate the potential development of supersensitivity at postsynaptic alpha 1-adrenergic receptors as a contributing factor to the exaggerated sympathetic response characteristic of autonomic dysreflexia. Adult male Wistar and Sprague-Dawley rats were anesthetized and given spinal transection at T5. After 30 days, rats were reanesthetized and arterial and venous catheters implanted. Twenty-four hours later, colorectal distension (CRD) was used to evoke autonomic dysreflexia in conscious, spinalized rats. To gauge changes in alpha 1-receptor sensitivity, we assessed mean arterial pressure (MAP) in response to intravenous phenylephrine (PE) infusions. No consistent differences were observed between intact and spinalized rats. Therefore, supersensitivity of alpha 1-receptors cannot completely account for the hypertensive bouts associated with autonomic dysreflexia. In addition, while attempting to develop an appropriate model for autonomic dysreflexia, we discovered that spinalized Wistar rats exhibited MAP responses characteristic of autonomic dysreflexia, whereas lesioned Sprague-Dawley rats did not, when subjected to CRD. Thus Wistar rats provide a better animal model for autonomic dysreflexia.

Adrenergic alpha-Agonists↗
PubMed1998

Peroxisome proliferator-activated receptors gamma and alpha mediate in vivo regulation of uncoupling protein (UCP-1, UCP-2, UCP-3) gene expression.

A role for peroxisome proliferator-activated receptors, PPAR gamma and PPAR alpha, as regulators of energy homeostasis and lipid metabolism, has been suggested. Recently, three distinct uncoupling protein isoforms, UCP-1, UCP-2, and UCP-3, have also been identified and implicated as mediators of thermogenesis. Here, we examined whether in vivo PPAR gamma or PPAR alpha activation regulates the expression of all three UCP isoforms. Rats or lean and db/db mice were treated with PPAR gamma [thiazolidinedione (TZD)] or PPAR alpha (WY-14643) agonists, followed by measurement of messenger RNAs (mRNAs) for UCP-1, UCP-2, and UCP-3 in selected tissues where they are expressed. TZD treatment (AD 5075 at 5 mg/kg x day) of rats (14 days) increased brown adipose tissue (BAT) depot size and induced the expression of each UCP mRNA (3x control levels for UCP-1 and UCP-2, 2.5x control for UCP-3). In contrast, UCP-2 and UCP-3 mRNA levels were not affected in white adipose tissue or skeletal muscle. Chronic (30 days) low-dose (0.3 mg/kg x day) TZD treatment induced UCP-1 mRNA and protein in BAT (2.5x control). In contrast, chronic TZD treatment (30 mg/kg x day) suppressed UCP-1 mRNA (>80%) and protein (50%) expression in BAT. This was associated with further induction of UCP-2 expression (>10-fold) and an increase in the size of lipid vacuoles, a decrease in the number of lipid vacuoles in each adipocyte, and an increase in the size of the adipocytes. TZD treatment of db/db mice (BRL 49653 at 10 mg/kg x day for 10 days) also induced UCP-1 and UCP-3 (but not UCP-2) expression in BAT. PPAR alpha is present in BAT, as well as liver. Treatment of rats or db/db mice with WY-14643 did not affect expression of UCP-1, -2, or -3 in BAT. Hepatic UCP-2 mRNA was increased (4x control level) in db/db and lean mice, although this effect was not observed in rats. Thus, in vivo PPAR gamma activation can induce expression of UCP-1, -2, and -3 in BAT; whereas chronic-intense PPAR gamma activation may cause BAT to assume white adipose tissue-like phenotype with increased UCP-2 levels. PPAR alpha activation in mice is sufficient to induce liver UCP-2 expression.

Adipose Tissue, Brown↗
PubMed1995

Convergence of multiple sensory inputs onto neurons in the dorsolateral medulla in cats.

The dorsolateral medulla, including the nucleus reticularis parvicellularis, the cuneate nucleus, and the external cuneate nucleus, is an integrative region for a variety of sensory inputs. The purpose of this study was to determine whether individual neurons respond to a variety of different sensory modalities. To this end, responses of 40 neurons in the dorsolateral medulla to multiple sources of sensory input were assessed in cats anesthetized with alpha-chloralose. Neurons were located in the nucleus reticularis parvicellularis (24 cells, 60%), the cuneate nucleus (10 cells, 25%), and the external cuneate nucleus (6 cells, 15%). All neurons were tested for responses to: electrical stimulation of afferents coursing through the left stellate ganglion and afferents in the left cervical vagus nerve, and somatic, auditory, and visual stimulation. No neurons responded to all five stimuli. Three cells (7.5%) responded to four stimuli, 11 (27.5%) responded to three stimuli, 10 (25.0%) responded to two stimuli, and 15 (37.5%) responded to only a single stimulus. The remaining cell was unresponsive to any stimulus. As a group, neurons in the nucleus reticularis parvicellularis received input from the greatest number of sensory modalities, and cuneate nucleus neurons received input predominantly from somatosensory afferents. External cuneate nucleus neurons displayed response profiles intermediate between nucleus reticularis parvicellularis and cuneate nucleus. In addition, eight neurons (20% of the total) were sensitive to changes in blood pressure. Results of the present study support the hypothesis that neurons in the nucleus reticularis parvicellularis receive convergent inputs from different sensory modalities.2+ behaviors.

Animals↗
PubMed1995

Phenylephrine and pilocarpine in the treatment of post-operative irido-corneal adhesion.

Following cataract surgery, entrapment of the iris within the surgical wound is often managed by intensive use of miotics. As the radial fibres stretch, only a small amount of fraction is exerted upon the entrapped iris. Application of a combination of phenylephrine and pilocarpine drops causes simultaneous contraction of the pupil sphincter and the radial muscle fibres. This study investigated the relative magnitude of forces induced in the iris periphery by pilocarpine and phenylephrine and the effectiveness of adding g. phenylephrine 10% to g. pilocarpine 4% drops in the treatment of postoperative irido-corneal adhesions. The investigation was divided into two parts. First, the forces induced in the iris periphery upon exposure to pilocarpine and phenylephrine were measured in 6 cadaver irises. The mean force was 27.5 +/- 5.7 x 10(-3) N for pilocarpine and 23.3 +/- 4.0 x 10(-3) N for phenylephrine. The combination of the two drugs produced a force of 54.2 +/- 6.6 x 10(-3) N (p < 0.05). In the second part of the study intensive pilocarpine 4% drops were administered to 17 patients who had iris-wound entrapment on the first post-operative day. Patients with persistent adhesion were commenced on intensive g. phenylephrine 10% and assessed after 90 minutes. Of the 17 patients, 6 responded to pilocarpine drops alone; in a further 7 the irido-corneal adhesion was released only by the addition of phenylephrine drops, and in 4 patients drops were ineffective in relieving the adhesion. This study indicates that addition of phenylephrine 10% to pilocarpine 4% drops enhances the effectiveness of pharmacological treatment of post-operative irido-corneal adhesion.

Cataract Extraction↗
PubMed1995

Sickle retinopathy in patients with sickle trait.

Sickle trait is traditionally considered a benign condition by ophthalmologists. Three cases of sickle retinopathy in subjects with sickle trait are reported. In all cases the onset of retinopathy was related to other contributing factors: in one case a traumatic hyphaema and raised intraocular pressure, in two others diabetes mellitus. Patients with sickle trait are at risk of retinopathy if coincident ocular or systemic disease is present.

Adolescent↗
PubMed1995

Auditing ophthalmology audits.

Medical audit is now a well-established part of the routine of ophthalmology departments. Accomplishing a complete audit is a complex task and its effectiveness depends on how it is performed. In order to evaluate the procedure in our department a retrospective analysis of 18 audits was carried out with particular reference to the level, objectives, beneficial effects, and effects on current practice. Our practice was critically analysed using these factors. One audit fulfilled the criteria for a full audit. Eleven (60%) were partial or potential and six (30%) were an attempt in planning an audit. The majority omitted plans for implementing change or targets for future assessment. Seven had educational benefit. There was little effect on patient care, cost-effectiveness or communication. Medical audit can be clinically effective, educational and stimulating. Failure to complete all stages affects the outcome. This study identifies the difficulties involved in performing complete audits and suggests improving the procedure by auditing audits on a regular basis.

Education, Medical↗
PubMed1994

Ophthalmologic features of thallium poisoning.

Thallium intoxication is characterized by the development of painful peripheral neuropathy, alopecia, mental disorders, and in severe cases, respiratory failure and death. Toxic optic neuropathy is also a feature. Ophthalmologic features of thallium poisoning include optic neuropathy, blepharoptosis, lens opacities, and ophthalmoplegia. A 44-year-old man with criminal sublethal thallium poisoning was examined one month after he was seen in the neurology department with classic systemic features. He was found to have diminished contrast sensitivity, a tritan defect in color vision, and a relative cecocentral scotoma before he developed optic atrophy.

Adult↗
PubMed1994

Lens hardness in mature cataracts.

A cataract is said to be mature when all the cortical fibres become opaque. Depending on the pathophysiological processes that cause the cortical fibre opacification this phenomenon may be associated with a varying degree of nuclear sclerosis. A relationship between lens hardness and degree of nuclear sclerosis in non-mature cataracts has been demonstrated previously. This study aimed to investigate the relationship between the hardness of mature cataracts and the transmitted nuclear colour, age and rate of progression of the cataract. Thirty-eight patients with mature cataracts were assessed prior to extracapsular cataract surgery. The nuclear colour that was transmitted through the opaque cortex was graded using reference photographs. Age and duration of visual symptoms were recorded and lens hardness was measured by a specially designed lens guillotine. Multivariate analysis of data indicates a relationship between hardness of a mature cataract and the transmitted nuclear colour and age (adjusted R2 = 0.59). There is also a tendency for hardening of the lens as the duration of visual symptoms increases. By considering these clinical markers, the cataract surgeon can estimate the hardness of the lens and therefore its suitability for phacoemulsification.

Age Factors↗
PubMed1994

Water content, lens hardness and cataract appearance.

Hardening of a cataractous lens is associated with the clinical appearance of brunescence and advancing age. Alterations in the nature and concentration of proteins, lens fibre compaction and other biochemical changes may all be contributing factors towards increased hardness. As the nucleus confers most of the rigidity to the lens and water content decreases towards the centre of the nucleus, the hardness might be thought to be related to total water content. To study the relationship between water content, hardness and the clinical appearance of cataract, 135 lenses were obtained from eyes undergoing extracapsular cataract surgery. The cataracts were assessed and classified pre-operatively. Lens hardness was determined by a specially designed guillotine and water content was measured by weighing the lens before and after desiccation. A regression analysis was carried out to look at the variation of lens water content with hardness, degree of nuclear sclerosis, extent of cortical and posterior subcapsular cataract, vacuolation and age. Multivariate analysis of data demonstrated a relationship between lens hardness and lens water content, degree of nuclear sclerosis and age (R2 = 0.59). An association could not be detected between lens water content and age, degree of nuclear sclerosis, extent of cortical and posterior subcapsular cataract and vacuoles. This study suggests that hardening of the lens is reflected by a respective decrease in water content. Increased hardness is associated with coloration and advancing age.

Adult↗
PubMed1994

Does topical flurbiprofen affect the pupillary response to acetylcholine?

Pre-operative topical non-steroidal anti-inflammatory drugs (NSAIDs) are known to be effective in maintaining pupillary dilation during cataract surgery. However, previous studies have suggested that pre-treatment with these agents interferes with the miosis produced by intraoperative acetylcholine injection. This study was designed to determine whether the pupillary response to acetylcholine injection is reduced following pre-operative topical flurbiprofen 0.03%. Pupil diameters were measured with a millimetre rule, and the measurements verified using computerised image analysis of digitised video images. Eyes treated with flurbiprofen were compared with controls. There was no significant difference in pupillary diameter before corneal incision, but following aspiration of soft lens matter and insertion of intraocular lens the pupil was significantly larger in the flurbiprofen group (p = 0.002). At 1.0, 2.5 and 5.0 minutes following injection of acetylcholine no significant difference in the degree of miosis was detected. This study confirms that topical flurbiprofen maintains dilation during cataract surgery, and does not find any evidence to suggest that intra-operative response to acetylcholine is reduced.

Acetylcholine↗
PubMed1993

The relationship between clinical classification of cataract and lens hardness.

'Hardness' of the cataractous lens is one of the major factors influencing the suitability of a patient for phacoemulsification. Surgeons should be able to estimate hardness when choosing patients for this method of cataract extraction. The aims of the study were to investigate the relationship between the clinical appearance of cataracts and their hardness. Ninety-one lenses were studied from patients undergoing endocapsular cataract extraction. Lenses were assessed using a classification based on the Lens Opacities Classification System II and hardness was assessed using an automated lens guillotine. The degree of nuclear sclerosis and the age of the patient were found to influence lens hardness. Regression analysis showed that 56% of the variation in hardness could be explained in terms of age and the degree of nuclear sclerosis. The presence of posterior subcapsular and cortical cataract did not influence hardness. These findings confirm our clinical suspicions and enable surgeons to make a more objective analysis of hardness prior to phacoemulsification.

Adult↗
PubMed1992

Mutation analysis of the Cys-X2-Cys-X19-Cys-X2-Cys motif in the beta subunit of eukaryotic translation initiation factor 2.

Recessive lethal mutations in the beta subunit of eIF-2 that restore HIS4 expression in the absence of an AUG start codon were isolated from diploid Saccharomyces cerevisiae strains. DNA sequence analysis of these alleles and of eIF-2 beta suppressor alleles isolated from haploid strains, identified point mutations that altered one of six amino acids that map to a Cys-X2-Cys-X19-Cys-X2-Cys "zinc finger" motif and immediately adjacent residues. Five of the affected amino acids are identical in the human and yeast eIF-2 beta protein. Together with earlier studies (Donahue et al., 1988), these point mutations implicate the zinc finger domain of eIF-2 beta in start-site selection during the scanning process. We have supplemented the mutations obtained by genetic selection with an additional set of constructed mutations in this region. Our studies indicate that the cysteine residues and the intervening amino acids of this motif are essential for eIF-2 beta function in translation initiation in vivo. However, the effects observed in cells containing a copy of eIF-2 beta with a deletion of this motif suggest that this mutated form is still able to associate with other components of the initiation complex, imparting defects on translation initiation. Thus, this motif may be required only for later events that lead to initiator codon recognition. Alterations in defined positions, as found in our suppressor alleles, could lead to recognition of non-AUG codons.

Alleles↗