Biomedical subjects

S J Hughes

Publications and source records attributed to S J Hughes.

At least 19 recordsLinked to original sources

PubMed1998

Lack of cell surface Fas/APO-1 expression in pulmonary adenocarcinomas.

The Fas receptor and ligand initiate an apoptotic pathway. Alterations in this pathway within tumor cells can result in escape from apoptosis and immune surveillance. We evaluated Fas protein expression in 42 primary pulmonary adenocarcinomas, and Fas expression and function in the lung adenocarcinoma cell lines A549 and A427. Immunohistochemical analysis demonstrated Fas protein expression in 47.6% of the tumors; however, Fas-positive tumors demonstrated cytoplasmic staining without cell surface expression. Northern blot analysis indicated that levels of Fas mRNA were similar in Fas protein-positive tumors to levels in normal lung tissue, but were reduced in Fas protein-negative tumors. Soluble form Fas was not detected in the majority of these tumors either by RT-PCR or Western blot analysis. Cell surface Fas protein expression was minimal in A549 and A427 cell lines as determined by flow cytometry. Both cell lines demonstrated Fas mRNA expression by Northern blot analysis and abundant protein expression by Western blot analysis. Transfection of the Fas cDNA derived from A549 cells induced surface Fas protein in COS cells; however, stable transfection of a native Fas cDNA into A549 cells failed to induce surface Fas protein expression. Parental A549 cells and A549 cells transfected with a Fas expression vector were resistant to Fas-mediated apoptosis. Transgenic expression of a FLAG-tagged Fas cDNA in A549 cells, with visualization of the Fas-FLAG protein using confocal microscopy, demonstrated that the Fas-FLAG protein was retained within cytoplasmic portions of the cell and was not translocated to the cell surface. These findings suggest that the Fas protein is reduced or not present on the cell surface in the primary lung tumors and is sequestered within A549 tumorigenic lung cells, and these alterations directly affect the cells resistance to Fas-mediated apoptosis.

Adenocarcinoma↗

PubMed1998

How effective is the Newport/Aspen collar? A prospective radiographic evaluation in healthy adult volunteers.

BACKGROUND: Cervical extrication collars are used in the evacuation of the injured to minimize secondary injury. These collars were designed for extrication and evacuation, not for longterm use, and complications have been reported when they have been used in the rehabilitation phase. The Newport/Aspen collar was designed specifically for long-term use. METHODS: Using a radiographic method, the effectiveness of the collar in restricting motion was evaluated for the cervical spine as a whole and for constituent segments, occiput to C7, in 15 normal volunteers. RESULTS: Combined flexion-extension was reduced from mean 98.8 to 31.1 degrees (31.5% of normal; p = 0.000000002), lateral bending was reduced from mean 31.1 to 15.9 degrees (51.1% of normal; p = 0.0000001), and overhead rotation was reduced from mean 64.6 to 26.8 degrees (41% of normal; p = 0.000000002). The cervical spine, however, does not move as one unit; paradoxical motion, the phenomenon of "snaking," occurs. Goniometric techniques do not demonstrate this effect. Data are provided for motion by segmental level. CONCLUSION: Full cervical immobilization is a myth. It would seem logical to match the level-specific efficacy of the device to the level of injury. Data are provided for the Aspen collar.

Adolescent↗

PubMed1998

Electrophysiological and metabolic characterization of single beta-cells and islets from diabetic GK rats.

We have used the whole-cell recording technique to determine whether ATP-sensitive potassium (K[ATP]) currents, voltage-dependent Ca2+ currents, and exocytosis are different in single beta-cells from pancreatic islets of Goto-Kakizaki (GK) rats, a novel model of NIDDM, and normal rats. In addition, we have also measured the insulin secretory responses, ATP content, and the rate of glucose metabolism in intact islets. Although the glucose sensitivity of the K(ATP) current was similar between GK rats and controls, in the absence of glucose, K(ATP) current density was larger in GK rats, which resulted in a more hyperpolarized membrane potential. Whole-cell Ca2+ currents were similar. By monitoring the cell capacitance with a fixed intracellular solution, no difference was detected in the exocytotic responses of beta-cells from normal and GK rats. In islets from GK rats, the rates of glucose utilization ([3H]H2O production from 5-[3H]glucose) and oxidation ([14C]CO2 production from U-[14C]glucose) were not significantly different from controls. Insulin secretion, however, was impaired (by 50%), and this was paralleled by a smaller increase in ATP content in response to stimulation by 10 mmol/l glucose in islets from GK rats when compared with controls. Under conditions in which K(ATP) channels were held open and the effects of glucose were independent of membrane potential, insulin release was still significantly lower in GK rat islets than in controls. These findings suggest that the impaired insulin secretion in islets from GK rats does not simply result from a failure to close K(ATP) channels, nor does it result from an impairment in calcium secretion coupling.

Adenosine Triphosphate↗

PubMed1997

Fas/APO-1 (CD95) is not translocated to the cell membrane in esophageal adenocarcinoma.

This study describes Fas (CD95) expression in Barrett's esophagus, adenocarcinomas of the esophagus, and three esophageal adenocarcinoma cell lines. Immunohistochemical analysis of Barrett's esophagus demonstrated cell surface expression of Fas protein. In contrast, 30.5% of esophageal adenocarcinomas examined by immunohistochemical analysis demonstrated faint cytoplasmic staining, and 69.5% were negative for Fas. Similar levels of Fas mRNA were identified in tumors compared to mRNA levels in esophageal squamous mucosa or Barrett's esophagus. An approximately Mr 48,000 Fas protein was identified by Western blot analysis in tumors that were negative for Fas expression by immunohistochemical analysis. The esophageal adenocarcinoma cell line Seg-1 was negative for Fas expression by immunohistochemical analysis, but Western blot analysis demonstrated abundant, appropriately sized Fas protein. In agreement with the immunohistochemical analysis, flow cytometry of Seg-1 showed minimal amounts of Fas on the cell surface, which correlated with resistance to Fas-mediated apoptosis. No mutations in the Seg-1 Fas coding sequence or exon 1 were identified by sequence analysis. This was confirmed by transient transfection of COS cells with expression vectors generated from the Seg-1 Fas cDNA, which resulted in cell surface expression of the Fas protein. Stable transfection of Seg-1 with a Fas expression vector did not result in efficient Fas expression on the cell surface. Seg-1 cells, transiently transfected with a Fas-FLAG expression vector and examined for protein expression using confocal microscopy and an anti-FLAG antibody, showed that the Fas-FLAG protein was not present on the cell surface but was present in the cytoplasm. Taken together, these results indicate that expression of Fas on the cell surface by esophageal adenocarcinoma is reduced. In an esophageal adenocarcinoma cell line, wild-type Fas protein is retained in the cytoplasm, and this correlates with resistance to Fas-mediated apoptosis. The retention of wild-type Fas protein within the cytoplasm may represent a mechanism by which malignant cells evade Fas-mediated apoptosis.

Adenocarcinoma↗

PubMed1997

Relaxin as a relaxant of the isolated rat uterus: comparison with its mechanism of action in vivo.

1. Glibenclamide, a blocker of ATP-sensitive potassium channels, has been shown to antagonize relaxin as a uterine relaxant in the rat in vivo but not in vitro. The aim, therefore, was to investigate whether the discrepancy between the two studies was a consequence of differences in (1) muscle layers, (2) hormonal conditions or (3) spasmogens utilized. Relaxin was compared with salbutamol and levcromakalim. 2. Relaxin was of similar potency as a uterine relaxant against oxytocin (0.2 mM)-induced spasm with tension measured in the circular or longitudinal muscle layers. Glibenclamide (10 microM) did not antagonize relaxin or salbutamol in these preparations but greatly antagonized levcromakalim (91-fold). Relaxin was a relaxant of tension activated by transmural electrical stimulation in uteri from rats that had been ovariectomized, although the maximal effect was only 30 +/- 15%, and in uteri from rats that had been treated with 17 beta-estradiol benzoate. Glibenclamide was not an antagonist of relaxin in the latter preparation but did antagonize levcromakalim (118-fold). Relaxin also inhibited spontaneous phasic tension development in uteri from ovariectomized rats but again was not antagonized by glibenclamide. 3. Because relaxin was not antagonized by glibenclamide under any of these various conditions, it would appear that the in vivo-in vitro discrepancy in the antagonism of relaxin by glibenclamide is not attributable to the effects of different muscle layers, hormonal conditions or spasmogens. It may be that the mechanism of action of relaxin or glibenclamide or both differs between in vivo and in vitro preparations.

Albuterol↗

PubMed1997

The role of a cAMP-dependent pathway in the uterine relaxant action of relaxin in rats.

The aim of this study was to investigate the role of the adenylyl cyclase pathway, and in particular cyclic AMP-dependent protein kinase A, in the relaxant action of relaxin in the isolated uterus of the nonpregnant rat. The purportedly selective inhibitor of cAMP-dependent protein kinase A N-[2-(methylamino) ethyl]-5-isoquinolinesulfonamide hydrochloride (H-8) (at 100 mumol l-7) antagonized relaxin, salbutamol (an agonist at beta-adrenoceptors) and levcromakalim (a K+ channel opener) to a similar extent (by factors of 3.1, 1.9 and 2.8, respectively), demonstrating that it is not a selective inhibitor. Relaxin and levcromakalim were less potent and had smaller, maximal, relaxant effects in longitudinal myometrium than in intact uterus cut in the longitudinal plane. By contrast, nifedipine (a Ca2+ channel blocker) was equipotent in the two preparations and salbutamol only slightly less potent in the longitudinal myometrium. Relaxin did not alter the cyclic AMP-dependent protein kinase A activity ratio in longitudinal myometrium, but did increase the activity ratio by a factor of 2.0 +/- 0.2 in the intact uterus. Salbutamol, the positive control, increased this activity ratio in both longitudinal myometrium (by 1.9 +/- 0.3 times) and in the intact uterus (by 3.8 +/- 0.3 times), whereas the negative control levcromakalim had no effect. Relaxin seems to act as a relaxant of longitudinal myometrium by a cyclic AMP-independent mechanism but possibly interacts with the circular myometrium or endometrium to release a relaxant factor via a cyclic-AMP-dependent mechanism.

Adrenergic beta-Agonists↗

PubMed1997

The effect of maternal protein deficiency during pregnancy and lactation on glucose tolerance and pancreatic islet function in adult rat offspring.

To test the hypothesis that poor foetal-neonatal nutrition predisposes adult animals to impaired glucose tolerance or diabetes, pregnant and lactating rats were fed a low (5%) protein diet and glucose tolerance and pancreatic islet function then assessed in the adult offspring. To expose any underlying defects the offspring were allowed access to a sucrose supplement (35%) or fed a high fat diet. Offspring born to low protein-fed females had significantly lower body weights than controls. In islets from previously malnourished rats, insulin release in batch incubations or perifusion was not significantly different to controls. In islets from previously malnourished animals fed sucrose, glucose-stimulated insulin release was reduced in perifusion by 66% (P < 0.01) and batch incubations by 26-52% (6-16 mmol/l glucose, P < 0.01). Similarly, impaired secretory responses were found in islets from previously malnourished animals fed a high fat diet. These did not result from a reduced pool of releasable insulin, as arginine-stimulated secretion was not impaired. Rats previously malnourished showed a normal glucose tolerance. Glucose tolerance was impaired, however, in previously malnourished rats fed sucrose (area under the glucose tolerance test curve was increased by 42%, P < 0.05) but despite the reduced islet secretory responses was not significantly different to sucrose-fed controls (area increased by 54%, P < 0.05). Glucose tolerance was impaired in previously malnourished animals fed high fat diet (area increased by 48%, P < 0.05) more so than in high fat fed-controls (28% increase, NS). These data support the hypothesis that poor foetal-neonatal nutrition leads to impaired pancreatic beta-cell function which persists into adult life. Alone this is not sufficient to produce diabetes, but an inability to respond to a highly palatable fat diet may tip the balance towards impaired glucose tolerance.

Animals↗

PubMed1996

Impaired glucose tolerance and mild hyperglycemia in sucrose-fed rats does not impair insulin secretion.

We fed normal rats a high sucrose diet in order to test the hypothesis that mild hyperglycemia can induce defects in pancreatic beta-cell function and impair glucose-stimulated insulin release. Rats provided with free access to a sucrose solution (35%) voluntarily consumed 50% more carbohydrate than control per day. After 7 days of sucrose feeding, glucose tolerance was significantly impaired; the area under the glucose tolerance test curve (GTT) was 683 +/- 61 mmol/120 min compared with 472 +/- 56 mmol/120 min in controls (P < 0.05). Impaired glucose tolerance was still present after a further 12 days (area under the GTT: 749 +/- 99 mmol/120 min). Sucrose-fed rats were significantly (P < 0.05) hyperglycemic by 1.5 mmol/l over controls. When insulin secretion was assessed in vivo and in vitro in control and sucrose-fed rats, no significant differences were apparent in plasma samples collected over a 1-h period or in statically incubated or perifused isolated pancreatic islets. In addition, the rates of glucose utilisation and oxidation were normal in islets from sucrose-fed rats. These data do not support the hypothesis that minimal hyperglycemia is sufficient to impair glucose-stimulated insulin release.

Animals↗

PubMed1996

The lack of a role for potassium channel opening in the action of relaxin in the rat isolated uterus; a comparison with levcromakalim and salbutamol.

1. The effects of relaxin in vitro in the isolated uterus from the non-pregnant rat were compared with those of levcromakalim and salbutamol in tissue bath, 42K+ -efflux and electrophysiological studies, to determine whether relaxin exhibits the characteristics of an opener of KATP-channels. 2. In uterus exposed to oxytocin (0.2 nM), tetraethylammonium (TEA, 10 mM) and glibenclamide (10 microM) produced large rightward shifts of the log10 concentration-effect curve to levcromakalim (125 fold and 118 fold, respectively). TEA (10 mM) caused only small rightward shifts of the log10 concentration-effect curves to salbutamol and relaxin (5.2 fold and 7.5 fold respectively). Glibenclamide did not antagonize salbutamol or relaxin. 3. Levromakalim (0.2 and 2 microM) suppressed the spasm evoked by low ( < or = 40 mM) but not high ( > 40 mM) concentrations of KCl. Salbutamol (1.5 nM) inhibited the spasm evoked by low concentrations of KCl ( < or = 40 mM). Salbutamol (15 nM) and relaxin (3 and 30 nM) inhibited the spasm evoked by low and high concentrations of KCl (10-80 mM). 4. Relaxin (0.12 microM) did not produce an increase in 42K+-efflux from longitudinal segments of rat myometrium. Exposure of tissues to relaxin (0.12 microM), in the presence of diltiazem (1 microM) plus KCl (20 mM), resulted in a small increase in 42K+-efflux of short duration. 5. Electrophysiological recording showed that the phasic spasms of the uterus exposed to oxytocin (0.2 nM) were accompanied by bursts of spiking activity superimposed upon a plateau potential. Inhibition of the mechanical activity of the uterus by levcromakalim (2 and 10 microM), salbutamol (30 nM) or relaxin (0.18 microM) was accompanied by a reduction in the duration of the plateau potential and the number of spikes without membrane hyperpolarization. 6. Unlike levcromakalim, relaxin did not selectively inhibit the spasm evoked by low concentrations of KCl and was not markedly antagonized by TEA or glibenclamide. Under conditions where a cromakalim-induced increase of the 42K+-efflux rate has been demonstrated, relaxin had only a very small effect. In isolated uterus from the rat, in contrast to observations in vivo, relaxin did not exhibit the characteristics of an opener of KATP-channels suggesting that another mechanism accounts for its inhibitory action.

Adrenergic beta-Agonists↗

PubMed1995

Insulin secretion from islets of GK rats is not impaired after energy generating steps.

Insulin secretory responses of intact and electrically permeabilised islets of Goto-Kakizaki (GK) rats, a novel model of non-insulin dependent diabetes mellitus, and Wistar (control) rats were compared to investigate the mechanism of the impairment of insulin secretion from pancreatic islets of GK rats. Insulin secretion from intact islets in response to glucose, glyceraldehyde, succinate monomethylester and tetramethyl p-phenylenediamine, which reduces cytochrome c directly, was significantly impaired in GK rats compared to control rats (P < 0.05, P < 0.01, P < 0.05 and P < 0.05, respectively). However, Ca(2+)-induced insulin release from electrically permeabilised islets of GK rats was higher than that of control rats. Moreover, insulin secretion from intact islets in response to 50 mM KCl, which depolarises islet cells, was not impaired in GK rats. These results indicate that insulin secretion from islets of GK rats is not impaired after energy generating steps of metabolism.

Animals↗

PubMed1995

Cellular localization of the inhibitory action of relaxin against uterine spasm.

1. The aim of this study was to determine whether the site of action of relaxin as a relaxant of rat myometrium is at the cell membrane or at an intracellular-site. Therefore, the potency of relaxin was determined against spasms reliant predominantly upon either extracellular Ca2+ or intracellular Ca2+. Uterine spasms dependent upon extracellular Ca2+ were elicited by (i) oxytocin (0.2 nM) (ii) Bay K 8644 (1 microM) in 10 mM K(+)-rich PSS and (iii) KCl (80 mM). Uterine spasm dependent upon intracellular Ca2+ was elicited by oxytocin (20 nM) in the presence of nifedipine (500 nM). The effects of relaxin against these spasmogens were compared with those of levcromakalim, nifedipine and salbutamol. 2. Relaxin (0.2-6.3 nM), levcromakalim (25-800 nM), salbutamol (1-63 nM) and nifedipine (1-250 nM) caused concentration-dependent inhibition of the spasm evoked by oxytocin (0.2 nM) and relaxin was the most potent relaxant. 3. Relaxin and nifedipine were slightly less potent against the spasm induced by Bay K 8644 (1 microM) than against spasm induced by oxytocin (0.2 nM) (15 fold and 13 fold respectively). Levcromakalim and salbutamol were equipotent against the spasm evoked by Bay K 8644 (1 microM) and that evoked by oxytocin (0.2 nM). 4. Relaxin induced only 47 +/- 7% inhibition of the KCl (80 mM)-evoked spasm at a concentration of 0.8 microM. Levcromakalim was much less potent (427 fold) against the spasm evoked by KCl (80 mM) than against the spasm evoked by oxytocin (0.2 nM). The potency of salbutamol against the spasm evoked by KCl (80 mM) was modestly reduced (14 fold) compared to that against the spasm evoked by oxytocin (0.2 nM). The potency of nifedipine against the KCl (80 mM)-evoked spasm was not different from that against the oxytocin (0.2 nM)-evoked spasm. 5. The potencies of relaxin and levcromakalim against the spasm evoked by oxytocin (20 nM) + nifedipine (500 nM) were greatly reduced (74 fold and 234 fold respectively) compared to their potencies against the spasm evoked by oxytocin (0.2 nM). The potency of salbutamol against these two spasmogens was not different. 6. Relaxin was much less potent against the spasm dependent upon intracellular Ca2+ (that induced by oxytocin (20 nM) + nifedipine (500 nM)) than against the spasms dependent upon extracellular Ca2+, those induced by oxytocin (0.2 nM) and Bay K 8644 (1 microM). In this regard, relaxin resembled levcromakalim and nifedipine rather than salbutamol. Therefore, the major site of action of relaxin appears to be located at the plasma membrane rather than at an intracellular level. The observation that relaxin was less effective against the KCl (80 mM)-induced spasm than against the oxytocin (0.2 nM)-evoked spasm may indicate that relaxin has a minor action involving K(+)-channel opening. 7. High concentrations of relaxin (up to 1 microM) induced significant inhibition of the spasm dependent upon intracellular Ca2+. Thus at high concentrations relaxin also appears to have an additional intracellular action.

Adrenergic beta-Agonists↗

PubMed1995

Developing an information systems strategy for nursing.

With the rapidly changing health care environment and information technology advances, organizations need to engage in strategic, planned change in order to allocate limited resources, achieve the organization's goals, and fulfill its mission [1]. One of the most important aspects of the organization's planned strategies for change concerns the information systems. The involvement of the nursing department in this process is critical. This poster presentation will communicate how nurses can develop an information systems strategic plan that will enable them to play an active role as contributors and vital participants in the strategic and business planning processes for information systems. This information systems strategy for nursing will: a) provide direction and purpose, b) guide nursing in identifying the kinds of information technology needed, c) assist in timely implementation of a system that supports nursing, and d) identify desired outcomes and benefits of an information system. The nursing information systems plan must be built on, and support, the organization's mission and business plan and integrate into the over-all information systems plans [2]. Components of the nursing strategic plan include the nursing mission statement and vision, an assessment of the current environment to identify supporting technology needed to achieve the nursing vision, expectations/anticipated outcomes, environmental considerations, and special staffing/expertise considerations. The nursing vision and mission statement is an articulation of the overall direction and purpose of the nursing organization. An assessment of the nursing organization, problem areas, opportunities for growth, the physical environment, existing systems, communications requirements, and resources is carried out to help identify areas where new technologies and automated methods of managing information could be applied. Special staffing and expertise not currently available in the organization, but necessary to the successful implementation of the plan, should be identified, and plans for filling those needs should be included in the planning and prioritization process. Based on the mission and assessment findings, goals or anticipated outcomes are developed. These goals must be realistic, financially feasible, and logistically achievable; they should also provide direction for action and decision-making [3]. Measurable objectives and detailed action plans can then be developed from these goals when implementation of this aspect of the strategic plan is begun. It is especially important, even at a strategic planning level, to consider change management techniques, including specific steps to involve individuals who will be affected by the change and to ensure open communication throughout the process. Efforts to collaborate with all affected departments and to offer input and educational opportunities to the various members of the health care team should be included in the strategic plan. A business plan describing the mission, goals, and objectives for a specific system implementation is the final step in the strategic planning process. The business plan includes expected outcomes and cost justification and may be done in cooperation with other departments (in the organization) that will be involved with this system. The business plan is used to communicate the information system's needs to the administration and governing board of the organization. With a good information systems strategy, nursing will be prepared to make more timely and better informed decisions related to applying information technology within the nursing department. The end results of this planning should be evident in the improved utilization of information technology to support the nursing vision and mission.

Arizona↗

PubMed1994

The role of islet secretory function in the development of diabetes in the GK Wistar rat.

Insulin secretion and glucose metabolism were compared in islets isolated from GK Wistar rats (a non-obese, spontaneous model of non-insulin-dependent diabetes mellitus) and control Wistars aged 8 and 14 weeks. By 8 weeks of age, GK Wistar rats were clearly diabetic as indicated by non-fasting plasma glucose concentrations and impaired glucose tolerance. Islet insulin content was not significantly different to controls at either age. In islets from 14-week-old GK Wistar rats glucose-stimulated insulin release (6-16 mmol/l glucose) was significantly reduced to 25-50% of controls in static incubations (p < 0.001). In perifusion, glucose-stimulated insulin release was reduced by 90% for first phase (p < 0.01) and by 75% for second phase (p < 0.05). The responses to arginine and 2 alpha Ketoisocaproate in islets were similar to those in controls. In contrast, islets isolated from 8-week-old GK Wistar rats exhibited no significant reduction in glucose-stimulated insulin secretion in static incubations. In perifusion, although both first and second phases of glucose-stimulated insulin release were slightly reduced, these were not significantly different to controls. Islets from 8-week-old GK Wistar rats failed however to respond to stimulation by glyceraldehyde. Raising the medium glucose concentration to 16 mmol/l significantly increased rates of glucose utilisation ([3H] H2O production from 5-[3H] glucose) and oxidation ([14C] CO2 production from U-[14C] glucose) in islets isolated from 8-week-old control and GK Wistar rats, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

PubMed1994

The role of reduced glucose transporter content and glucose metabolism in the immature secretory responses of fetal rat pancreatic islets.

Isolated fetal islets show an immature or poor secretory response to nutrient secretagogues which may result from impaired mitochondrial oxidative processes. Insulin secretion, glucose metabolism and detection of metabolic enzymes by radiolabelling and immunoprecipitation were compared in islets isolated from neonatal (aged 5 days) and fetal rats (at 20 days gestation). The insulin secretory dynamics of fetal islets were abnormal in response to stimulation by glucose (10 mmol/l); a rapid release of insulin reaching a maximum 6 min after stimulation was observed with no rising second phase release. However, when the data were expressed as percentage of islet insulin content released, fetal islets released significantly more insulin than neonatal islets in response to glucose (4.86 +/- 0.45% vs 1.81 +/- 0.62%, p < 0.01) or 100 nmol/l glibenclamide (2.49 +/- 0.17% vs 0.25 +/- 0.06%, p < 0.001). Fetal islets however, failed to release insulin in response to stimulation by glyceraldehyde (10 mmol/l) unlike neonatal islets. Both glucose utilisation (as measured by the formation of [3H] H2O from 5-[3H] glucose) and glucose oxidation (as measured by the formation of [14C] CO2 from U-[14C] glucose) did not increase significantly in response to increasing the medium glucose concentration to 10 mmol/l whereas in neonatal islets, glucose utilisation and glucose oxidation were significantly increased 2.5- and 2.7-fold, respectively. When islets were incubated with both radiolabelled glucoses simultaneously, the rate of glucose oxidation was shown to be directly proportional to the rate of glucose utilisation. The relationship between glucose utilisation and glucose oxidation was similar in fetal and neonatal islets.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Characterization of Ca2+/calmodulin-dependent protein kinase in rat pancreatic islets.

We have attempted to identify islet Ca2+/calmodulin-dependent protein kinase (CaM kinase) by comparing its activity with purified brain CaM kinase II. Islet CaM kinase, in the presence of calmodulin and Ca2+, phosphorylated major endogenous substrates of 102, 57 and 53 kDa and also exogenous glycogen synthase; brain CaM kinase II phosphorylated glycogen synthase and peptides of 57 and 53 kDa. Alloxan (1 mM) inhibited the phosphorylation of glycogen synthase and the 102, 57 and 53 kDa islet peptides by islet CaM kinase; the phosphorylation of glycogen synthase and the 57 and 53 kDa substrates by brain CaM kinase II was also inhibited by alloxan. The Ca2+ and calmodulin-dependencies of phosphorylation of the endogenous islet substrates differed. In the presence of 400 nM calmodulin, half-maximal phosphorylation was attained at Ca2+ concentrations of 80 +/- 9, 401 +/- 61 and 459 +/- 59 nM for the 102, 57 and 53 kDa substrates respectively. In the presence of 10 microM Ca2+, half-maximal phosphorylation was attained at calmodulin concentrations of 9 +/- 2, 38 +/- 2.5 and 37 +/- 2 nM for the 102, 57 and 53 kDa substrates respectively. Differential centrifugation located the 102 kDa substrate in the post-100,000 g supernatant and the 57 and 53 kDa substrates in the particulate fraction. These data suggest that islet CaM kinase is similar to, if not identical with, brain CaM kinase II, but that phosphorylation of the endogenous 102 kDa substrate occurs by a distinct kinase which shows different sensitivities to Ca2+ and calmodulin. This kinase probably corresponds to CaM kinase III and the 102 kDa peptide to elongation factor 2 (EF-2), since the 102 kDa peptide was shown to undergo ADP-ribosylation in the presence of diphtheria toxin and NAD+.

Adenosine Diphosphate↗

PubMed1993

Chronic bile diversion does not alter canine interdigestive myoelectric activity.

Previous studies have suggested that the cyclic entry of bile into the duodenum during fasting regulates interdigestive patterns of motility by releasing the putative regulatory hormone motilin. Our aim was to determine if cyclic secretion of bile into the duodenum regulates interdigestive myoelectric activity and plasma motilin concentrations. Six dogs were prepared with gastric and intestinal serosal electrodes. Myoelectric activity was measured during fasting and after a meal before and after reoperative translocation of the entrance of the bile duct to the mid-jejunum. The characteristics of the migrating myoelectric complex (MMC) and conversion to a postprandial pattern were similar before and after bile duct translocation. The period (112 +/- 5 vs 109 +/- 10 min; mean +/- SEM), migration velocity of phase III through the duodenum (8.9 +/- 1.2 vs 6.8 +/- 0.5 cm/min), and duration of individual phases of the MMC in the stomach, duodenum, and jejunum were not altered significantly (each P > 0.05) by chronic diversion of bile from the duodenum. Plasma motilin concentrations were similar before and after bile duct translocation (P > 0.05), continued to cycle temporally with the MMC, and peak concentrations occurred during phase III and were greater than during phases I and II (P < 0.01). We conclude that the presence of bile in the lumen of the duodenum does not regulate interdigestive myoelectric patterns of the canine upper gut or the cyclic release of motilin.

Animals↗

PubMed1993

Length of hospital stay and outcome after femoral neck fracture: a prospective study comparing the performance of two hospitals.

Length of hospital stay and outcome after femoral neck fracture were compared in a prospective study between two adjacent hospitals. In matched populations, mean length of stay was 30.8 days at Hospital X and 15.7 days at Hospital Y. Need for rehousing, age over 80 years and new nursing home placement prolonged length of stay at Hospital X, but not at Hospital Y. Hospital X had an orthopaedic rehabilitation ward and returned 88 per cent of patients to their own home, placing 9 per cent admitted from home in nursing homes. Hospital Y returned 76 per cent of patients to their own home and 19 per cent to nursing homes. The rapid discharge policy of Hospital Y saved significant resources within the acute hospital at the expense of returning significantly fewer patients to their own homes.

Aged↗

PubMed1993

Inhibition of contractions of the isolated human myometrium by potassium channel openers.

OBJECTIVE: The aim was to investigate the properties of two potassium channel openers in human myometrium. STUDY DESIGN: The abilities of aprikalim and BRL 38227 to inhibit contractions produced by potassium chloride and oxytocin of myometrial strips from nonpregnant and pregnant women were studied. RESULTS: Aprikalim (1 and 10 mumol/L) and BRL 38227 (1 and 10 mumol/L) suppressed contractions induced by low (10 and 20 mmol/L) but not high (40 and 80 mmol/L) potassium chloride concentrations. Aprikalim and BRL 38227 were also potent relaxants of oxytocin (1 nmol/L)-induced contractions with rapid onset of action, of similar potency to each other, and reversible. Both compounds were significantly more potent against oxytocin (1 nmol/L)-induced contractions in myometrial strips from nonpregnant than from pregnant women. Aprikalim and BRL 38227, as relaxants of oxytocin-induced contractions, were antagonized by glibenclamide (1 mumol/L), a blocker of adenosine 5'-triphosphate-sensitive potassium channels. CONCLUSIONS: The results suggest that aprikalim and BRL 38227 relax the human myometrium by potassium channel opening, possibly at adenosine 5'-triphosphate-sensitive potassium channels. Compounds of this pharmacologic class are useful for investigating the physiologic functions of potassium channels in this tissue and could have a place in the treatment of dysmenorrhea and preterm labor.

Benzopyrans↗