Body mass index and occupational accidents among health care workers in a large university hospital.

INTRODUCTION: Obesity is associated with a number of chronic diseases such as cardiovascular diseases and cancers. The association of obesity with occupational accidents has been suggested although the evidence is less convincing. The objective of the study is to analyse the relationship between BMI values and ergonomic accidents in a large University Hospital. METHODS: The relationship between body mass index (BMI) and the incidence of ergonomic occupational accidents over a period of 8 years was investigated in a cohort of employees of a large University Hospital, covering almost 27,000 person-years of observation. This relationship was stratified according to the variables age, gender, functional status within the organization and work schedule (part-time or full time). Height and weight were objectively measured, demographic data were obtained from the human resource department and the registration of ergonomic accidents was carried out by the safety and prevention department of the hospital. RESULTS: The number of ergonomic accidents, expressed as number/1000 person-years was higher for female employees compared to male employees, increased with age and markedly increased from functional class A (leading or expert function and higher educational level) to D (executive function in patient care and technical department). However, the incidence of ergonomic accidents accompanied by loss of working time was not significantly associated with BMI, independently of age and gender. In addition, the type of accident and the severity of the accidents expressed as the number of days absent from work were unrelated to BMI. CONCLUSION: No independent relationship between BMI and the incidence of ergonomic accidents could be identified in our cohort. Tailoring working conditions to individual BMI levels is not recommended.

The drone ambulance [A-UAS]: golden bullet or just a blank?

Defibrillation within the first minutes after sudden cardiac arrest can save many quality-adjusted life years. Yet, despite enormous investments, 'healthcare' is still unable to provide this for the majority of patients. Emergency Medical Services often have a too long mean response time and many issues surround Public Access Defibrillation programs. In this article we argument that AED-equipped drones could be the 'magic bullet'. They are easily deployed and fast, and have a relatively low operational cost. As such they could rapidly bring an AED next to the victim, irrespective of most geographical circumstances, give visual feedback and situational awareness to the EMS dispatcher and thus assist a bystander to provide better CPR. Although there are many real-life barriers to actual deployment, we argument these might all get solved once we have solved the described technological issues.

Influence of aging on the beta- and glucagon-receptor-mediated glycogenolysis in rat hepatocytes.

The influence of aging on beta-receptor and glucagon-receptor control of glycogenolysis was investigated in rat hepatocytes. The beta-receptor-induced glucose output was detectable only in senescent rats, was partly dependent on extracellular Ca2+, and was inhibited by 4 beta-phorbol 12-myristate 13-acetate (PMA), insulin, and the Ca(2+)-antagonists, verapamil and nifedipine. Chelation of extracellular Ca2+ potentiated the effect of nifedipine only. In contrast, glucagon-stimulated glycogenolysis, similar in mature and senescent rats, was independent on extracellular Ca2+ and was unaffected by PMA. Verapamil, in senescent rats only, and nifedipine, in mature and senescent rats, inhibited glucagon-stimulated glucose output only in the presence of Ca2+. Insulin inhibited glucagon-induced glucose output, irrespective of the age of the rat and the presence of Ca2+. We conclude that the beta-receptor component in the adrenergic regulation of glycogenolysis in senescent rats consists of a major Ca(2+)-independent and a minor Ca(2+)-dependent part, displaying different sensitivity towards protein kinase C (PKC), Ca(2+)-antagonists, and insulin. Aging does not change the capacity of glucagon to induce a full glycogenolytic response in the absence of extracellular Ca2+; Ca(2+)-influx, however, seems to be involved when extracellular Ca2+ is present, and this sensitivity is increased on aging.

Desensitization of alpha 1-, beta- and glucagon receptors in rat hepatocytes: influence of ageing.

The alpha 1-agonist phenylephrine in hepatocytes from mature and senescent rats, and the beta-agonist isoproterenol in hepatocytes from senescent rats, elicited a time-dependent, homologous desensitization of alpha 1- and beta-receptor-mediated glycogenolysis respectively, which was maximal after 20 min of exposure to the agonists. Glucagon triggered desensitization of the glycogenolytic response to glucagon, isoproterenol and phenylephrine, which was maximal after less than 5 min; this was followed by resensitization of the glucagon response only. After 20 min of treatment with phenylephrine or isoproterenol, no change in cellular distribution of alpha 1- or beta-receptors was noticed, using sucrose gradient centrifugation. After a 1-h exposure to both agonists, a shift of both receptors to a light density fraction was found, reflecting receptor internalization. Neither the rate of functional desensitization, nor the degree of receptor internalization was altered upon ageing. We conclude that functional desensitization and internalization of adrenergic receptors in rat hepatocytes are separate events in time and are largely unaffected by the ageing process. Thus, despite the absence of a beta-receptor-mediated glycogenolytic response in hepatocytes from mature rats, isoproterenol triggers the internalization of beta-receptors.

Use of elastase as an internal standard in immunoblotting techniques.

A non-radioactive method for relative, semi-quantitative analysis of immunoblots, based on the use of elastase as internal standard and conventional peroxidase staining was devised and applied to the immunoassay of Gs-proteins in crude membrane preparations of rat kidneys. We found that the coefficients of variation of samples, run within the same experiment or run within different experiments, are reduced to half or a quarter of their original value respectively when corrected for elastase as an internal standard, allowing meaningful comparison of these samples.

Receptor function in heart failure.

In patients with congestive heart failure, down-regulation of beta-adrenoceptors is present, probably as a result of sympathetic overstimulation. In end-stage dilated cardiomyopathy, beta 1-adrenoceptor density is markedly reduced, while beta 2-adrenoceptor density is normal. This latter finding does not necessarily imply normal sensitivity to beta 2-stimulation, due to possible alterations in the beta-adrenoceptor/adenylate cyclase complex beyond the receptor. In some disease states, such as ischemic cardiomyopathy and mitral valve disease, there seems to be a concomitant reduction of the beta 1- and beta 2-adrenoceptor density. The finding of beta-adrenoceptor down-regulation has stimulated the search for novel therapeutic approaches in heart failure patients. Beta-agonists could even further down-regulate beta receptors, and this perhaps explains why they seem not to be useful in long-term use. Agents that directly stimulate adenylate cyclase activity, such as forskolin, or that increase cyclic adenosine monophosphate degradation, such as the phosphodiesterase inhibitors, are being tested. Beta-adrenoceptor blocking agents were used in treatment of heart failure before beta-adrenoceptor down-regulation was recognized in these patients. It is tempting to speculate that the beneficial clinical and hemodynamic effects seen in these patients treated with metoprolol is indeed due to an antagonism of the beta-adrenoceptor down-regulation. Studies testing whether beta-adrenoceptor blocking agents can improve survival in congestive heart failure patients are on-going.

Distinction between surmountable and insurmountable selective AT1 receptor antagonists by use of CHO-K1 cells expressing human angiotensin II AT1 receptors.

1. CHO-K1 cells that were stably transfected with the gene for the human AT1 receptor (CHO-AT1 cells) were used for pharmacological studies of non-peptide AT1 receptor antagonists. 2. In the presence of 10 mM LiCl, angiotensin II caused a concentration-dependent and long-lasting increase of inositol phosphates accumulation with an EC50 of 3.4 nM. No angiotensin II responses are seen in wild-type CHO-K1 cells. 3. [3H]-Angiotensin II bound to cell surface AT1 receptors (dissociates under mild acidic conditions) and is subject to rapid internalization. 4. Non-peptide selective AT1 antagonists inhibited the angiotensin II (0.1 microM) induced IP accumulation and the binding of [3H]-angiotensin II (1 nM) with the potency order: candesartan > EXP3174 > irbesartan > losartan. Their potencies are lower in the presence of bovine serum albumin. 5. Preincubation with the insurmountable antagonist candesartan decreased the maximal angiotensin II induced inositol phosphate accumulation up to 94% and, concomitantly, decreased the maximal binding capacity of the cell surface receptors. These inhibitory effects were half-maximal for 0.6 nM candesartan and were attenuated by simultaneous preincubation with 1 microM losartan indicating a syntopic action of both antagonists. 6. Losartan caused a parallel rightward shift of the angiotensin II concentration-response curves and did not affect the maximal binding capacity. EXP3174 (the active metabolite of losartan) and irbesartan showed a mixed-type behavior in both functional and binding studies. 7. Reversal of the inhibitory effect was slower for candesartan as compared with EXP3174 and irbesartan and it was almost instantaneous for losartan, suggesting that the insurmountable nature of selective AT1 receptor antagonists in functional studies was related to their long-lasting inhibition.

Activation of phospholipase C by cholecystokinin receptor subtypes with different G-protein-coupling specificities in hormone-secreting pancreatic cell lines.

Phospholipase C (PLC) activity was investigated by stimulation of membrane preparations obtained from insulin (beta-TC3)-, somatostatin (Rin 1027-B2)-, and glucagon (INR1-G9)-producing pancreatic cell lines using the non-hydrolyzable GTP analogue GTPgammaS alone, the C-terminal octapeptide cholecystokinin (CCK-8), or gastrin. All compounds caused a significant 2- to 4.4-fold stimulation of PLC activity in the different cell lines, which was diminished by the non-hydrolyzable GDP analogue GDPbetaS. CCK receptor subtypes were characterized by radioligand binding experiments. High-affinity binding sites for tritiated CCK(A) receptor antagonist L-364,718 (K(d) = 0.24 nM) and tritiated CCK(B) receptor antagonist L-365,260 (K(d) = 0.13 nM) were only present in Rin 1027-B2 cells. High-affinity binding sites for both ligands were not found in beta-TC3 or INR1-G9 cells. Competition binding experiments with non-labeled CCK receptor antagonists CR 1505 (CCK(A) receptor-selective) and CR 2945 (CCK(B) receptor-selective), as well as microphysiometry experiments, resulted in the same receptor distribution. Reverse transcriptase-polymerase chain reaction confirmed the CCK receptor distribution pattern for Rin 1027-B2 cells, but in addition showed the existence of CCK(B) receptors in beta-TC3 cells. Immunoblocking experiments with C-terminal antibodies against different G-protein alpha-subunits demonstrated inhibition of CCK-stimulated PLC activity in beta-TC3 cells by G(q/11)alpha antiserum (70%), in Rin 1027-B2 cells by G(q/11)alpha antiserum (70%) and G(i)-3alpha antiserum (23%), and in INR1-G9 cells by G(q/11)alpha antiserum (60%) and G(o)alpha antiserum (45%). We conclude that CCK receptor subtypes with different G-protein-coupling specificities to PLC are present in the different hormone-secreting cells of the endocrine pancreas.

Effect of age on beta-receptors, Gs alpha- and Gi alpha- proteins in rat heart.

beta-adrenoceptors, Gs alpha- and Gi alpha-proteins were investigated in a crude plasma membrane preparation from ventricles of young (2-4 months) and senescent (22-24 months) Wistar rats. Receptor density, ligand affinity and beta 1/beta 2-receptor ratio were independent of the age of the rats. The percentage of beta-receptors coupled to G-proteins increased with age. An age-related increase in the level of Gs alpha (124%) was paralleled by an increase in the ratio between the high and low molecular weight form of Gs alpha. The level of Gi alpha-protein almost doubled (170%) upon aging. We conclude that the age-related differences are small at the level of the beta-adrenoceptor molecule, but that the increase in Gi alpha-proteins could be responsible for the age-related reduction in myocardial inotropic and chronotropic responses. Moreover, we suggest that the changes in degree of high affinity coupling between beta-receptor and Gs-protein are possibly linked to alterations in the ratio between the Gs-molecular weight subtypes.

Characterization of the beta-adrenergic transduction system in spleen mononuclear leukocyte membranes of young and senescent rats.

The effect of aging on some of the properties of the beta-adrenergic transduction system was determined in a membrane fraction of spleen mononuclear leukocytes from young (2-3 months) and old (24-25 months) rats. Receptor density was unchanged and the percentage of receptors in the high affinity configuration for isoproterenol was reduced with increasing age. Adenylate cyclase activity, either unstimulated or stimulated with forskolin, GTP or isoproterenol was not affected by aging. This suggests the presence of compensatory mechanisms in the beta-adrenergic signal transduction system of rat spleen mononuclear cells in order to ensure equal cAMP production for equal agonist stimulation.

Pharmacological characterization of a beta 3-receptor agonist (BRL 37,344) and a partial agonist (CGP 12,177A) in neonatal rat liver plasma membranes.

The pharmacological properties of BRL 37,344 (sodium-4-(2'-[2-hydroxy-2- (3-chloro-phenyl)ethylamino]-propyl)phenoxyacetatesesquihydrate), a beta 3-selective agonist, and CGP 12,177A) (-)-4-(3-t-butyl amino-2-hydroxypropoxy) benzimidazole-2-one], a non-selective beta-antagonist, recently characterized as a partial beta 3-agonist in rat adipose tissue, were studied in comparison with isoproterenol, a non-selective beta-agonist, in plasma membranes prepared from the livers of newborn rats. Competition binding curves obtained with [125I]iodocyanopindolol ([125I]CYP) as ligand and isoproterenol or BRL 37,344 as competitor were characterized by the presence of a high and a low affinity binding site; the high affinity binding site was no longer detectable when guanidylimidobisphosphate (GppNHp) was present in the incubation mixture. Competition curves with CGP 12,177A were monophasic and independent of GppNHp. In the presence of 10(-7) M of the beta 2-selective antagonist ICI 118,551 [erythro-(+/-)-1-(7-methylindan-4-yloxy)-3-isopropylamino butan-2-ol], a concentration which blocks most of the beta 2-receptors, ligand binding was reduced to 32% of its maximum. Under these conditions, isoproterenol further displaced the ligand, and competition curves still displayed the high and the low affinity binding sites; BRL 37,344, however, caused no further displacement of ligand, except at the highest concentrations. This suggests that BRL 37,344 occupies only the ICI 118,551-sensitive binding sites, i.e. beta 2-receptors. Isoproterenol and BRL 37,344 both stimulated adenylate cyclase (EC 4.6.1.1) activity concentration dependently, although the stimulating effect of BRL 37,344 was about half of what was found for isoproterenol. Furthermore, BRL 37,344 inhibited concentration dependently the isoproterenol-induced stimulation of adenylate cyclase, and the inhibition was dependent on the concentration of isoproterenol. The stimulating effect of isoproterenol and BRL 37,344 on adenylate cyclase was blocked by ICI 118,551, whereas the beta 1-selective antagonist CGP 20,712A ((+/-)-(2-(3-carbamoyl-4-hydroxyphenoxy)-ethylamino)-3-[4-(1-methy l-4- trifluoromethyl-2-imidazolyl)-phenoxy]-2-propanolmethane sulphonate) was ineffective. CGP 12,177A failed to stimulate adenylate cyclase activity. From these results we suggest that BRL 37,344 acts as a beta 2-partial agonist in rat liver. The results obtained with CGP 12,177A are typical for a non-selective beta-antagonist. We therefore conclude that there is no pharmacological evidence for the presence of beta 3-receptors in livers from newborn rats.

Subcellular localization of neuronal nitric oxide synthase in rat small intestine.

The subcellular localization of neuronal nitric oxide synthase (NOS I, EC 1.14.13.39) was investigated in the longitudinal muscle/myenteric plexus (LM/MP) preparation of rat small intestine. The presence of NOS I, inducible nitric oxide synthase (NOS II), and endothelial nitric oxide synthase (NOS III) was assessed after homogenization and low-speed centrifugation in a postnuclear supernatant by immunological detection after PAGE and Western blotting. Only NOS I was clearly present, whereas NOS II and NOS III were below detection limits. After high-speed centrifugation of the postnuclear supernatant, soluble and particulate fractions were obtained, and the presence of NOS I in these fractions was investigated by measurement of NOS I immunoreactivity and enzyme activity. We found that 90 +/- 1% of NOS I immunoreactivity and 97 +/- 1% of NOS enzyme activity were confined to the soluble fraction of the tissue. Further immunological analysis demonstrated that washing the particulate fraction revealed detectable amounts of NOS I only after concentration of the washing supernatant. Most particulate NOS I remained in the pellet and therefore represents cell organelle-associated enzyme. No NOS I immunoreactivity could be detected as a soluble protein within organelles of the cell. Particulate NOS I could in part be solubilized by Triton X-100 treatment, and the detection of Triton X-100-soluble NOS I was dependent on the antibody used. In conclusion, our results indicate that NOS I in the LM/MP preparation of rat small intestine is mainly soluble and that the particulate NOS I is partly an intrinsic membrane protein and can partly be solubilized by detergent treatment.

Effect of maturation and aging on beta-adrenergic signal transduction in rat kidney and liver.

The characteristics of the beta-adrenergic signal transduction system were analyzed in kidney and liver membrane preparations from neonatal (2-3 days), mature (2 months), and old (2 years) rats. When comparing kidneys from adult to neonatal rats, we found a higher beta-receptor density and a higher percentage of beta(1)-receptor subtype, lower immunoreactive G(salpha)-protein, a lower ratio between the high and low molecular weight splice variant of G(salpha), lower immunoreactive G(ialpha)-protein, and lower basal adenylate cyclase activity. When comparing livers from adult to neonatal rats, we found lower beta-receptor density and basal adenylate cyclase activity. Very few differences could be detected when comparing mature to old kidneys or livers. Stimulated adenosine 3',5'-cyclic monophosphate (cAMP) synthesis was tissue- and age-dependent. In liver, G-protein- and beta-receptor-stimulated cAMP synthesis mirrored basal adenylate cyclase activity and was highest in liver from neonatal animals. In contrast, cAMP synthesis was significantly more stimulated in kidneys from mature animals than from neonatal and senescent rats. We conclude that: (i) the stoichiometry of the components within the beta-receptor/G-protein/adenylate cyclase complex is not fixed but is both tissue- and age-dependent; (ii) adenylate cyclase enzyme activity is possibly but not necessarily the rate-limiting step in the beta-receptor-mediated synthesis of cAMP; and (iii) there is in vivo evidence for a preferential co-expression of the large splice variant of the G(s)-protein and beta(2)-receptor subtype. It is speculated that this could have important physiological consequences for the development of the kidney.

Thermodynamic analysis of isoproterenol binding to beta-adrenoceptors in rat lung membranes.

The thermodynamic properties of the binding of the beta-adrenoceptor agonist isoproterenol and of the antagonist propranolol to beta-adrenoceptors of rat lung were investigated. We found that in our experimental conditions, the high- and low-affinity binding sites for the agonist displayed different properties: the binding to the high-affinity binding site was entropy-driven with a small increase in enthalpy, while agonist binding to the low-affinity binding site was enthalpy-driven. Binding of isoproterenol in the presence of GTP or its non-hydrolyzable analogue GppNHp, and the binding of propranolol were enthalpy-driven with a small increase in entropy.

Effect of aging on properties and function of beta-adrenoceptors in rat lung.

beta-Adrenoceptor density and ligand affinity, basal adenylate cyclase activity and cAMP synthesis upon stimulation with forskolin, fluoride, guanine nucleotides (GTP or guanylyl imidodiphosphate (GppNHp) or isoproterenol in the presence of the nucleotides were studied in membranes prepared from lungs of young (aged 2-3 months) and of old (aged 24-25 months) male Wistar rats. There was a significant (P less than 0.05, 21%) increase in beta-receptor density and a significant (P less than 0.05, 38%) decrease in the percentage of high-affinity binding sites for isoproterenol. Both basal adenylate cyclase activity and that after stimulation with guanine nucleotides or isoproterenol in the presence of nucleotides were unaltered with age. Forskolin stimulation of cAMP synthesis was significantly reduced (by 24%, P less than 0.05) in tissues from older animals. It is suggested that the age-dependent changes in properties of beta-receptors in rat lungs are compensatory, in order to ensure equal cAMP production for equal agonist stimulation.

The beta-adrenergic transduction system in kidneys from young and senescent rats.

beta-Adrenoceptor density, ligand affinity, high-affinity agonist binding, basal adenylate cyclase activity and cAMP synthesis upon stimulation with either forskolin, F-, guanine nucleotides (GTP or GppNHp) or isoproterenol in the presence of the nucleotides were studied in membranes prepared from kidneys of young (2-3 month) and old (24-25 month) male Wistar rats. There is a significant (P less than 0.01) 62% increase in beta-receptor density, a significant (P less than 0.05) 115% decrease in ligand affinity, a significant (P less than 0.05) 33% decrease of high-affinity binding sites for (-)-isoproterenol and a significant (P less than 0.01) 151% decrease of the affinity of the high-affinity agonist binding site. Basal adenylate cyclase activity and the activity after stimulation with guanine nucleotides and forskolin were significantly higher in old animals as compared to young (P less than 0.01). Stimulation of the system with isoproterenol in the presence of GTP was more effective in old animals, although the P less than 0.05 level of significance was barely reached. It is suggested that age-dependent changes of the beta-adrenoceptors in rat kidney are similar to those described for lungs: changes at the different levels of the beta-adrenergic transduction chain associated with age are compensatory so as to ensure equal cAMP synthesis for a given agonist stimulation.

Human neuropeptide YY1 receptors exert unequal control of the extracellular acidification rate in different cell lines.

The ability of the human neuropeptide YY1 receptor subtype to increase the extracellular acidification rate in different cell lines was investigated by using the Cytosensor Microphysiometer. In CHO-Y1 cells (Chinese Hamster Ovary cells expressing the cloned human neuropeptide YY1 receptor), neuropeptide Y increased the acidification rate by up to 15% of the basal level with a -Log(EC50) of 7.42. As expected for neuropeptide YY1 receptors, this response was potently inhibited by the neuropeptide YY1-selective non-peptide antagonist BIBP3226 ((R)-N2-(diphenylacetyl)-N-[(4-hydroxy-phenyl)methyl]-D-arginine amide). Its enantiomer BIBP3435 ((S)-N2-(diphenylacetyl)-N-[(4-hydroxy-phenyl)methyl]-D-arginin amide) was less potent. The antagonists themselves did not affect the extracellular acidification rate at concentrations up to 10 microM. In SK-N-MC cells (a neuroblastoma cell line of human origin that expresses the neuropeptide YY1 receptor) no change of the acidification rate could be observed in the presence of neuropeptide Y at concentrations up to 1 microM. For control, the neuropeptide YY1 receptors were also investigated by assessing whole cell radioligand binding and, at the functional level, by assessing their ability to decrease the forskolin-induced accumulation of cAMP. The specific (i.e., neuropeptide Y-displaceable) binding of [3H]neuropeptide Y was to a homogeneous class of high-affinity sites in both SK-N-MC and CHO-Y1 cells. The equilibrium dissociation constants for [3H]neuropeptide Y, the total number of binding sites and the kinetic constants for association and for dissociation were similar. Neuropeptide Y produced a dose-dependent inhibition of forskolin-induced cAMP accumulation in SK-N-MC cells (-log(EC50) = 9.40) but it did not affect cAMP accumulation in CHO-Y1 cells. Non-transfected CHO-K1 cells were used as negative control throughout the study. No binding or response could be observed in these cells. Our data suggest that the signalling mechanisms of neuropeptide YY1 receptors are closely related to the cell type in which they are expressed.

Changes in the expression of alpha(2)-adrenergic receptor subtypes during maturation of neuronal cells from fetal pig superior cervical ganglia.

The expression of presynaptic alpha(2)-adrenergic receptor (alpha(2)-AR) subtypes was investigated in cultured neurons from fetal pig superior cervical ganglion (SCG). Cells were incubated with chicken antibodies against alpha(2)A-, alpha(2)B- or alpha(2)C-AR subtypes either alone or together with antibodies against dopamine-beta-hydroxylase (DbetaH, a marker for adrenergic neurons) or against choline acetyl transferase (ChAT, a marker for cholinergic neurons). We found immunoreactivity for all three alpha(2)-AR subtypes in SCG-cells when cultured for 8-11 days. The relative expression of the alpha(2)A-subtype was approximately 1/3 of that of alpha(2)B- and alpha(2)C-AR. Co-localisation of all three alpha(2)-AR subtypes was observed in cells expressing DbetaH or ChAT. Increasing the potassium concentration in the culture medium increased the expression of DbetaH and decreased the expression of the alpha(2)A- and alpha(2)C-subtype without altering the expression of the alpha(2)B-subtype. Co-culture of neurons with pig splenocytes enhanced the expression of ChAT and decreased the expression of the alpha(2)B-subtype without altering the expression of alpha(2)A- and alpha(2)C-subtypes. Our results indicate that the three alpha(2)-receptor subtypes are expressed on both noradrenergic and cholinergic nerves. Induction of the noradrenergic phenotype favours the expression of the alpha(2)B-subtype over that of the alpha(2)A- and alpha(2)C-subtype. Conversely, enhancement of the cholinergic phenotype favours the expression of the alpha(2)A- and alpha(2)C-subtypes over that of the alpha(2)B-subtype. Our results suggest that the alpha(2)B-receptor is preferentially associated with noradrenergic nerve endings.

Glycan microheterogeneity of alpha 1-acid glycoprotein in sera and dialysates of patients on continuous ambulatory peritoneal dialysis.

Total concentration and concanavalin (Con A) dependent microheterogeneity of alpha 1-acid glycoprotein (AGP) were studied in sera of eight chronic renal failure patients before the start of continuous ambulatory peritoneal dialysis (CAPD), and in sera and dialysate fluids for up to 6 months of CAPD. The glycan heterogeneity of AGP in the samples was expressed as a reactivity coefficient, i.e. the ratio of the Con A-reactive AGP components to the Con A non-reactive AGP component. Concentrations of AGP in serum and reactivity coefficients were markedly elevated in non-dialysed uraemic patients. AGP concentrations in serum increased further during the first week on CAPD, and then gradually decreased to pre-dialysis values, which were reached after 1 to 6 mth. The reactivity coefficients did not change significantly during the CAPD treatment. Dialysate AGP concentrations were low in comparison to those in serum, and there was a good correlation between the reactivity coefficients in the dialysate fluids and those in the corresponding sera. The effect of peritonitis was evaluated in a separate group of eight CAPD patients. Serum and dialysate AGP concentrations were significantly higher during than after peritonitis while the corresponding reactivity coefficients were only slightly elevated.

G-proteins in kidneys of spontaneously hypertensive rats.

G(s alpha)-, total G(i alpha)- and G(q/11alpha)-protein concentrations were investigated by quantitative immunoblotting in membranes of total kidney, renal cortex and medulla as well as in cortical tubules and glomeruli of Spontaneously Hypertensive Rats (SHR) and normotensive Wistar Kyoto rats (WKY), aged 5 weeks, 3 or 8 months. We found that total kidney of 5 week old SHR possess less G(s alpha)-, G(i alpha)- and G(q/11alpha)-proteins than controls. For G(s alpha)-proteins, differences found in total kidney were mirrored both in cortex (tubules and glomeruli) and in medulla. Decreased G(i alpha)-concentrations were accompanied by lower tubular but higher glomerular levels, while medullar levels were also increased. Decreased G(q/11alpha)-concentrations were reflected in decreased glomerular and medullary concentrations. Kidneys of 3 month old SHR and WKY possessed similar concentrations of all G(alpha)-species. In 8 month old SHR similar G(i alpha)-, but decreased G(s alpha)-and G(q/11alpha)-concentrations were observed. The G(s alpha)-decrease was reflected in cortex and medulla, the G(q/11alpha)-decrease in the medulla. We conclude that the main strain-related differences in G(alpha)-concentrations are seen in prehypertensive SHR.