Investigations into the absorption of insulin and insulin derivatives from the small intestine of the anaesthetised rat.

Experiments have been undertaken to determine the extent to which cholic acid conjugates of insulin were absorbed from the small intestine of anaesthetised rats by means of the bile salt transporters of the ileum. The measure used to assess the absorption of the cholyl-insulins was the amount of hypoglycaemia following infusion into the small intestine. Control experiments involving infusion of natural insulin into the ileum showed either nil absorption or absorption of a small amount of insulin as indicated by transient dip in the blood glucose concentration. However, when insulin was co-infused with the bile salt taurocholate, this was followed by a marked hypoglycaemic response which was specific to the ileum and did not occur on infusion into the jejunum. When the two cholyl conjugates of insulin were tested viz. B(29)-Lys-cholyl-insulin and B(1)-Phe-cholyl-insulin, both were biologically active as indicated by hypoglycaemic responses on systemic injection, though their potency was about 40% of that of natural insulin. While there was no evidence for the absorption of B(29)-Lys-cholyl-insulin when infused into the ileum, B(1)-Phe-cholyl-insulin did cause a long lasting hypoglycaemic response, indicating that absorption had occurred. Since the hypoglycaemic response was blocked on co-infusion with taurocholate and was absent for infusion of the conjugate into the jejunum, these results were taken as evidence that B(1)-Phe-cholyl-insulin had been taken up by the ileal bile salt transporters. This would indicate that B(1)-Phe-cholyl-insulin is worthy of further investigation for use in an oral insulin formulation.

Development and evaluation of thin-film flexible microelectrode arrays for retinal stimulation and recording.

We have described the development of a flexible microelectrode array with potential applications in the large scale recording of neural signals and in focal electrical stimulation for use as a prosthetic implant in degenerative retinal diseases. The array under test consisted of 61 platinum electrodes of 5 µm diameter with 60 µm spacing connected by 8 µm wide gold tracks encased in a flexible polyimide substrate of 15 µm thickness from which recordings were taken from 16 electrodes. The device was tested on an exposed frog eyecup preparation which is characterized by small retinal ganglion cells of similar dimensions to those present in the human retina. The responses of these cells evoked by photic stimulation consisted of trains of action potentials of high signal-to noise ratio at each of the recording sites. Delivery of cathodal constant voltage pulses and constant current pulses to specific electrodes in the array led to the generation of action potentials in adjacent electrodes, implying that retinal ganglion cells in the proximity had been stimulated. Since prolonged stimulation with supra-threshold voltages impaired neither electrode structure nor retinal function, these results provide a sound basis for scaling up the number of array electrodes to deliver focal electrical pulses to the retina, as would be required by a viable epiretinal prosthesis.

Lack of Restoration in Vivo by K-Channel Modulators of Jejunal Fluid Absorption after Heat Stable Escherichia coli Enterotoxin (STa) Challenge.

Enhanced potassium ion permeability at the enterocyte basolateral membrane is assumed to facilitate sustained chloride ion and fluid secretion into the intestinal lumen during episodes of secretory diarrhoeal disease. To examine this concept in vivo, two potassium ion channel blockers and a channel opener were coperfused with E. coli heat stable STa enterotoxin to determine whether such compounds improved or worsened the inhibited fluid absorption. In the STa (80 ng/mL) challenged jejunal loop, the fluid absorption rate of 28.6 ± 5.8 (14) µL/cm/hr was significantly below (P < .001) the normal rate of 98.8 ± 6.2 (17) µL/cm/hr. Intraluminal (300 uM) glibenclamide added to STa perfused loops failed to improve the inhibited fluid absorption rate, which was 7.4 ± 3.2 (6) µL/cm/hr on coperfusion with STa. Similarly, on coperfusion with 30 uM clotrimazole, the fluid absorption rate with STa present remained inhibited at 11.4 ± 7.0 (4) µL/cm/hr. On coperfusion with intraluminal 1 uM cromakalim, STa reduced fluid absorption significantly (P < .02) to 24.7 ± 8.0 (10) µL/cm/hr, no different from STa challenge in the absence of cromakalim. Infusion i.v. with these agents also failed to restore fluid absorption after STa challenge. These observations do not support the proposed potassium ion permeability event as a necessary corollary of enterotoxin-mediated secretion. This makes it unlikely that modulators of such permeability prevent enterocyte secretion in diarrhoeal disease.

Effects of uncertainty and target displacement on the latency of express saccades in man.

Saccadic eye movements generated in response to a gap paradigm in which the fixation light spot was extinguished 200 ms prior to presentation of the target light spot showed appreciably shorter latencies than for the overlap paradigm in which the target light spot was presented 200 ms prior to extinction of the fixation light spot. When there was unpredictability in the direction of target presentation, i.e., to the left or right of the fixation light spot, the gap paradigm evoked mainly fast regular saccades of peak latency of 155 ms with relatively few express saccades which were defined as having latencies of less than 120 ms. By contrast, when the target always appeared to the right, a substantial population of express saccades with peak latency 95 ms was now generated. There was also a change in the relationship between saccadic latency and target angular displacement which covered the range 5-35 degrees . With the overlap paradigm and unpredictability of target direction, the latencies of the slow regular saccades increased markedly with target angular displacement. This was not the case with the same target direction when the latency of slow regular, fast regular, and express saccades remained constant with increasing target angular displacement. This indicates for targets appearing in the same hemifield that the ocular motor system operates with shortest latency irrespective of target angular displacement.

Relationship between visual field loss and contrast threshold elevation in glaucoma.

BACKGROUND: There is a considerable body of literature which indicates that contrast thresholds for the detection of sinusoidal grating patterns are abnormally high in glaucoma, though just how these elevations are related to the location of visual field loss remains unknown. Our aim, therefore, has been to determine the relationship between contrast threshold elevation and visual field loss in corresponding regions of the peripheral visual field in glaucoma patients. METHODS: Contrast thresholds were measured in arcuate regions of the superior, inferior, nasal and temporal visual field in response to laser interference fringes presented in the Maxwellian view. The display consisted of vertical green stationary laser interference fringes of spatial frequency 1.0 c deg(-1) which appeared in a rotatable viewing area in the form of a truncated quadrant extending from 10 to 20 degrees from fixation which was marked with a central fixation light. Results were obtained from 36 normal control subjects in order to provide a normal reference for 21 glaucoma patients and 5 OHT (ocular hypertensive) patients for whom full clinical data, including Friedmann visual fields, had been obtained. RESULTS: Abnormally high contrast thresholds were identified in 20 out of 21 glaucoma patients and in 2 out of 5 OHT patients when compared with the 95% upper prediction limit for normal values from one eye of the 36 normal age-matched control subjects. Additionally, inter-ocular differences in contrast threshold were also abnormally high in 18 out of 20 glaucoma patients who had vision in both eyes compared with the 95% upper prediction limit. Correspondence between abnormally high contrast thresholds and visual field loss in the truncated quadrants was significant in 5 patients, borderline in 4 patients and absent in 9 patients. CONCLUSION: While the glaucoma patients tested in our study invariably had abnormally high contrast thresholds in one or more of the truncated quadrants in at least one eye, reasonable correspondence with the location of the visual field loss only occurred in half the patients studied. Hence, while contrast threshold elevations are indicative of glaucomatous damage to vision, they are providing a different assessment of visual function from conventional visual field tests.

Absorption of the cholic acid-conjugated peptide hormone cholylsecretin from the rat ileum in vivo.

AIMS: Previously, we demonstrated that gastrin peptides as long as 34 amino acids were absorbed from the ileum of rat after conjugation to the C24 position of cholic acid and that these peptides retained full biological activity. As absorption was specific to the ileum, it was inferred that the conjugated hormone was taken up by the bile salt transporters. We have now extended these experiments to a member of a different family of hormones, viz. secretin, a 27-amino acid hormone that stimulates serous secretions from the exocrine pancreas. METHODS: After conjugation to cholic acid, the degree of cholylsecretin absorption from the ileum of anaesthetized rats was assessed from the increase in pancreatic secretions. RESULTS: A complication to the study was that intra-ileal infusion of native secretin caused a transient increase in the levels of pancreatic secretions. This was in contrast to the effects of intra-ileal infusion of cholylsecretin which did not cause this transient increase but, instead, gave rise to a delayed increase in pancreatic secretions which was sustained over several hours during which cholylsecretin was detected in plasma in high concentration by mass spectrometry. The pancreatic response to cholylsecretin was abolished by co-infusion of 50 mm taurocholate, employed to compete with the bile salt transporters, although a transient increase in pancreatic secretions similar to that caused by secretin was now generated. This was shown to arise from an action of taurocholate per se causing the release of endogenous secretin which is present in rat ileum. CONCLUSIONS: We, therefore, concluded that cholylsecretin had been absorbed from the rat ileum by uptake by bile salt transporters.

Absorption of biologically active peptide hormones from the small intestine of rat.

Absorption of the 4, 10 and 34 amino acid forms of gastrin from the small intestine has been investigated in anaesthetized rats. The method of assessment of successful absorption of the hormone into the systemic circulation was when the amount of acid secreted by the stomach over consecutive 15-min periods was increased. When the natural hormones were infused into the ileum in a relatively high dose, there was no increase in gastric acid secretion, indicating that they had not been absorbed. Each of the forms of gastrin was conjugated at the free amino terminus to the carboxyl group of cholic acid. Subsequent infusion of the conjugated form of gastrin into the ileum, this time in relatively low doses, resulted in substantial and prolonged increases in gastric acid secretion, indicating that these hormones had been successfully absorbed. In addition, conjugation of the 10 and 34 amino acid forms of gastrin with cholic acid was shown to increase markedly the potency in evoking an increase in gastric acid secretion in response to intravenous injection of the hormone. Absorption of the gastrin conjugates was specific to the ileum thus indicating that they had been absorbed through the bile salt transporters.

The dependence of binocular contrast sensitivities on binocular single vision in normal and amblyopic human subjects.

We have measured monocular and binocular contrast sensitivities in response to medium to high spatial frequencies of vertical sinusoidal grating patterns in normal subjects, anisometropic amblyopes, strabismic amblyopes and non-amblyopic esotropes. On binocular viewing, contrast sensitivities were slightly but significantly increased in normal subjects, markedly increased in anisometropes and esotropes with anomalous binocular single vision (BSV) and significantly reduced in esotropes and exotropes without BSV. Application of a prismatic correction to the strabismic eye in order to achieve bifoveal stimulation resulted in a significant reduction in contrast sensitivity in esotropes with and without anomalous BSV, in exotropes and in non-amblyopic esotropes. Control experiments in normal subjects with monocular viewing showed that degradative effects of the prism occurred only with high prism powers and at high spatial frequencies, thus establishing that the reduced contrast sensitivities were the consequence of bifoveal stimulation rather than optical degradation. Displacement of the image of the grating pattern by 2 deg. in normal subjects and anisometropes by a dichoptic method to simulate a small angle esotropia had no effect on the contrast sensitivities recorded through the companion eye. By contrast, esotropes showed similar reductions in contrast sensitivity to those obtained with the prism experiments, confirming a fundamental difference between subjects with normal and abnormal ocular alignments. The results have thus established a suppressive action of the fovea of the amblyopic eye acting on the companion, non-amblyopic eye and indicate that correction of ocular misalignments in adult esotropes may be disadvantageous to binocular visual performance.

An analysis of the dependence of saccadic latency on target position and target characteristics in human subjects.

BACKGROUND: Predictions from conduction velocity data for primate retinal ganglion cell axons indicate that the conduction time to the lateral geniculate nucleus for stimulation of peripheral retina should be no longer than for stimulation of central retina. On this basis, the latency of saccadic eye movements should not increase for more peripherally located targets. However, previous studies have reported relatively very large increases, which has the implication of a very considerable increase in central processing time for the saccade-generating system. RESULTS: In order to resolve this paradox, we have undertaken an extended series of experiments in which saccadic eye movements were recorded by electro-oculography in response to targets presented in the horizontal meridian in normal young subjects. For stationary or moving targets of either normal beam intensity or reduced red intensity, with the direction of gaze either straight ahead with respect to the head or directed eccentrically, the saccadic latency was shown to remain invariant with respect to a wide range of target angular displacements. CONCLUSIONS: These results indicate that, irrespective of the angular displacement of the target, the direction of gaze or the target intensity, the saccade-generating system operates with a constant generation time.

Polyurethane gel liner usage in the Oxford Prosthetic Service.

The objective was to investigate which lower limb amputees are using Alpha polyurethane gel liners and the effects of these on comfort and suspension of their prosthesis. A retrospective study was carried out of case records of all patients issued with Alpha cushion and locking liners between 1997 and the end of January 1999. The type of liner used was compared with age, sex, level and cause of amputation, time since amputation, comfort and suspension. Modified Stanmore/Harold-Wood mobility grades; duration of use and number of liners issued per patient were recorded. Sixteen (16) patients were identified who had been prescribed Alpha cushion liners. Improved comfort was reported by all. Forty (40) patients were identified who had been prescribed Alpha locking liners. Twenty (20) of these reported improved comfort and 10 improved suspension. The average time since amputation was 18.5 years for those using cushion liners and 14.1 years for locking liner users. Fifty-two (52) of all 56 patients using Alpha cushion and locking liners had mobility grades of 4 or more. Trauma was the most common cause of amputation. This group is a relatively mobile group of amputees. All those using cushion liners reported improved comfort. Some of the locking liner users reported improved comfort and suspension but this was not universally the case.

Evolution of the perioperative clinical nurse specialist role.

Perioperative nursing roles continue to rapidly change as we enter the twenty-first century. The need for strong leadership skills, expert clinical skills, creative management, ongoing continuing education, and research continues to grow in every department of surgical services. The clinical nurse specialist plays an important role in addressing each of these needs. Great opportunities exist within the field of nursing for perioperative nurses to expand their practice using their creativity, ideas, and skills. Using the clinical nurse specialist in the perioperative setting can foster creativity, stimulate development of new methods based on research, and maximize the delivery of high quality care by the entire OR staff.

Effect of ethanol on myocardial infarct size in a canine model of coronary artery occlusion-reperfusion.

This study was conducted to determine if elevated blood alcohol prior to acute coronary artery occlusion affects myocardial infarct size in an in vivo canine model. Seven pentobarbital anesthetized open-chest dogs received 10 min iv infusion of ethanol (0.08 g/kg/min). Ten min after ethanol, the left anterior descending coronary artery (LAD) was occluded distal to its first major branch for 60 min. The LAD was then reperfused for 5 h. Following electrically induced ventricular fibrillation, the area at risk of infarction was delineated with dye. The area of infarction was identified by staining with triphenyl tetrazolium chloride. Eleven untreated control experiments were also conducted. Mean blood ethanol concentration was 155+/-26 mg/dl just prior to LAD occlusion and 47+/-3 mg/dl after 4 h reperfusion. Ethanol infusion had no effect on systemic hemodynamic variables during ischemia. In ethanol treated animals, the area at risk was 19.7+/-3.0% of the left ventricle, and the infarct size was 20.9 +/-4.8% of the area at risk. In control experiments, the area at risk was 23.0+/-4.1% of the left ventricle (p > 0.05), and the infarct size was 21.6+/-3.8% of the area at risk (p > 0.05). Collateral blood flow to ischemic region did not differ between the two groups, and the relationships between infarct size and collateral flow were similar for control and untreated hearts. Acute ethanol exposure prior to coronary artery occlusion and subsequent reperfusion does not affect myocardial infarct size in the heart of the anesthetized dog.

The population of users of upper limb prostheses attending the Oxford Limb Fitting Service.

There is no central database that records the changing provision of prostheses in the United Kingdom. Experience suggests there have been some shifts in the population, particularly in the past decade. Because the detailed records of these changes are contained in the patients' medical records it is difficult to assess the substance of these data except on an individual basis; the larger picture the sifting of a centre's or many centres' data. This paper describes the analysis of one such set of records at the Oxford Limb Fitting Centre. It relates the profile of the population that attends the centre to the general population, and compares the information with that obtainable from other sources. The possible causes for the findings are discussed.

An investigation into the relative merits of pituitary adenylate cyclase-activating polypeptide (PACAP-27) and vasoactive intestinal polypeptide as vagal neuro-transmitters in exocrine pancreas of rats.

Pancreatic exocrine secretions were collected over 15 min periods and analysed in terms of weight of juice, total HCO3- and total protein in anaesthetized and pithed rats. Pituitary adenylate cyclase-activating polypeptide (PACAP) (i.v.) evoked a serous HCO3- secretion which contained relatively little protein, together with a marked vasodepressor action. The latter was still maximal at lower doses of PACAP, which evoked diminished pancreatic secretions. The effects of PACAP were similar to those evoked by the same dose of VIP and by cervical vagal stimulation, while secretion evoked a much larger secretion of fluid and HCO3-. The time courses of the PACAP-evoked secretions were significantly delayed compared with those of VIP. In the pithed rat, PACAP caused the same level of pancreatic secretions as in the anaesthetized rat, though this was now accompanied by a substantial pressor response which was blocked by phentolamine or prazosin, indicating that it was alpha 1-adrenoceptor mediated. VIP caused a depressor response in the pithed rat, as well as the same level of pancreatic secretions as in the anaesthetized rat. The putative VIP antagonist [Lys1,Pro25,Arg3,4,Tyr6]-VIP (abbreviated as VIPi) caused a selective and significant reduction in the HCO3- secretion evoked by VIP and blocked the vasodepressor response caused by VIP. By contrast, VIPi did not antagonize either the secretory or vasodepressor actions of PACAP. Unilateral electrical stimulation of the cervical vagus nerve evoked significant increases in the weight of juice, total protein and total HCO3- secreted. When preceded by injection of VIPi, vagally evoked secretions were unchanged in terms of weight of juice and total protein but had a significantly reduced HCO3- content. These results are consistent with the release of VIP, though not PACAP, as a vagal neurotransmitter in the exocrine pancreas.

The optical quality of the human eye revisited.

A subjective assessment of the optical quality of the human eye may be derived from a comparison between the contrast sensitivities obtained in response to a refracted grating display and a non-refracted grating display. The former may be generated on a cathode ray tube (CRT) while the latter may be obtained with laser interference fringes observed in the Maxwellian view. With this method, the rate of decline of the contrast ratio of the optical media with increasing spatial frequency has been shown to be appreciably less than the rate of decline of the neural performance represented by the laser contrast sensitivities. However, this comparison has been based on results in response to a green CRT display and red (Helium-Neon) interference fringes. In the present study, we have confirmed this result with respect to comparisons between a green CRT display and a green laser display, although the absolute value of the contrast ratio of the optical media for the green laser display was reduced compared with values for the red laser display.

Invariance of the pattern electroretinogram evoked by psychophysically equivalent stimuli in human ageing.

1. The aim of this study was to assess the contribution of retinal ganglion cells to the decline in contrast sensitivity during human ageing. 2. After determination of the appropriate refraction for each subject, younger subjects were arranged to be exposed to a display luminance which was suprathreshold by the same amount as in older subjects wearing a 4.0 mm diameter artificial pupil with a neutral density filter. 3. In fifty-four subjects, aged 20-99 years, contrast sensitivities measured in response to phase-reversed grating patterns of 2, 5 and 8 cycles per degree declined significantly with increasing age at each spatial frequency studied. 4. Subjects were made psychophysically equivalent by setting the display contrast at x5 and x10 contrast threshold for each subject. The pattern electroretinogram (PERG) was recorded with a sterile silver thread (DLT) electrode placed in the lower canthus of one eye, with the indifferent electrode on the temple and the earth on the forehead. 5. For each contrast multiple at each spatial frequency, the PERG implicit time showed no significant change with age, indicating equivalence of the response across the age range. 6. Control experiments with two young and two elderly subjects established that the PERG implicit time decreased appreciably with increasing contrast, over a range of x2 to x20 contrast threshold. 7. Since the psychophysically equivalent stimulus displays had generated equivalent PERGs in terms of implicit time in young and elderly subjects, this was consistent with the equivalence of retinal ganglion cell function under these conditions. 8. Adverse changes within the retina were therefore inferred to play a major role in the decline in contrast sensitivity with age.

Rehabilitation outcome 5 years after 100 lower-limb amputations.

Received wisdom commends a policy of maximizing the ratio of below-knee to above-knee amputations in patients with end-stage arterial disease. After adoption of this policy, the long-term outcome of 100 consecutive lower-limb amputations in 96 patients was monitored by annual review for 5 years. The ratio of primary below-knee to above-knee amputations was 2:1, with 9 per cent of below-knee amputations undergoing revision to a higher level. At 2 years after amputation only 26 per cent of patients were successfully walking out of doors, while 40 per cent had died. By 5 years 67 per cent were dead and only 9 per cent continued to walk out of doors with an artificial limb, although a further 8 per cent continued to use the limb within the confines of their own homes. In a previous audit of 193 amputations performed during the 3.5 years to December 1984, stump healing was a problem in 45 per cent of primary below-knee amputations, compared with 25 per cent in the present study. Although the below- to above-knee ratio in 1984 was only 1:2, the overall rehabilitation rate, as determined by the proportion of patients able to walk at 2 years, was 34 per cent. It is concluded that increasing the proportion of below-knee amputations from one-third to two-thirds of lower-limb amputations for occlusive arterial disease does not improve effective rehabilitation rates. Received wisdom on the desirability of a high below- to above-knee ratio may be wrong.

Enhancement by atropine of the pancreatic exocrine secretions evoked by vagal stimulation in the pithed rat.

1. Pancreatic secretions were collected in response to 15 min periods of bilateral stimulation of the cervical vagus nerves in the pithed rat. 2. The weight of juice, total HCO3- and total protein evoked by a second period of vagal stimulation were essentially similar to those of the first period of vagal stimulation. 3. When the second period of vagal stimulation was preceded by an intravenous bolus injection of atropine sufficient to block the vagally induced bradycardia, the weight of secretion and total protein were greatly potentiated over an extended time course far exceeding that of the period of vagal stimulation. Total HCO3- was unchanged. 4. By contrast, atropine was effective in antagonizing the stimulatory effects of the muscarinic agonist methacholine injected intravenously. 5. The putative VIP (vasoactive intestinal polypeptide) antagonist [D-p-chloro-Phe6, Leu17]-VIP injected intravenously also increased the vagally evoked weight of juice, with total HCO3- and total protein unchanged. This was explicable by a partial agonist effect which was additive to the stimulatory action of vagal stimulation. 6. To explain these results, it is proposed that endogenously released acetylcholine exerts a negative feedback effect on the postganglionic varicosities which release both acetylcholine and another cotransmitter which was not excluded as being VIP. In the presence of atropine, the cotransmitter is proposed to be released from the inhibitory feedback, thus enhancing the response to vagal stimulation.

Changes in visual function with normal ageing, cataract and intraocular lenses.

We have been able to separate the neural and optical components of vision with contrast sensitivity tests employing laser interferometry and direct viewing of a cathode ray tube display. In normal ageing, neural function declined markedly while optical function remained essentially unchanged. On the other hand, even the mildest degree of cataract was shown to degrade optical function to the extent that it exceeded the age-related neural deterioration. The satisfactory optical performance of the monofocal implant lens has been confirmed, though the diffractive bifocal implant lens was shown to have optical limitations. A major cause of these was shown, by simulation experiments, to be the dilution of the contrast of the in-focus image by the superimposed defocused image, particularly under conditions of reduced retinal sensitivity as would be present in the elderly.

Quality of vision through diffractive bifocal intraocular lenses.

Two elderly women have each received a monofocal intraocular lens in one eye and a 3M diffractive bifocal intraocular lens in the other eye. Both eyes were shown to have equivalent retinal/neural function by measuring contrast sensitivity to laser interference fringes which bypassed refractive and other defects of the ocular media. The eyes with a bifocal intraocular lens displayed a much greater depth of focus, though at the expense of diminished contrast sensitivity compared with the normal values expected for that age. Simulation experiments suggested that the observed reduction in contrast sensitivity was not adequately explained by a simple reduction in retinal illumination of the in-focus image as might intuitively be expected from the bifocal separation of incident light to two simultaneous focal points. The simultaneous superimposition of the out-of-focus image on the in-focus image must also be considered, since this caused a significant reduction in contrast sensitivity when the retinal illumination was insufficiently above the photopic luminance threshold.