Biomedical subjects

B Johansen

Publications and source records attributed to B Johansen.

At least 37 records · Page 2Linked to original sources

PubMed1999

Activation of protein kinase A by dibutyryl cAMP treatment of NIH 3T3 cells inhibits proliferation but fails to induce Ser-133 phosphorylation and transcriptional activation of CREB.

The cAMP analogue dibutyryl cAMP (dbcAMP) is often used to activate the protein kinase A pathway and to study the expression of cAMP-responsive genes. Here we show that in NIH 3T3 cells dbcAMP is able to activate PKA, but fails to stimulate expression of the cAMP-inducible c-fos gene. Co-expression of A-kinase anchoring protein 75, previously shown to amplify cAMP signalling and to stimulate c-fos expression, could not restore cAMP responsiveness of the c-fos promoter. DbcAMP-induced activation of PKA may result in poor translocation of the catalytic sub-units of PKA to the nucleus, indicated by the lack of both Ser-133 phosphorylation of the cAMP-response element binding factor CREB and stimulation of the transcriptional activity of this factor. DbcAMP treatment, however, inhibited cell proliferation. These results suggest that cAMP-mediated inhibition of proliferation may be independent of translocation of the catalytic sub-units into the nucleus.

3T3 Cells↗

PubMed1999

Concerted expression of BK virus large T- and small t-antigens strongly enhances oestrogen receptor-mediated transcription.

Previous studies have shown that the human polyomavirus BK (BKV) genome contains an oestrogen response element (ERE). This isolated element binds its cognate receptor in vitro and can mediate 17beta-oestradiol-induced gene expression when linked to a heterologous promoter. The roles of the ERE- and the AP-1-binding sites in oestrogen receptor-directed transcription from the complete BKV promoter/enhancer (Dunlop strain) have been examined and the effects of the general co-activator CBP and large T- and small t-antigens on oestrogen receptor-mediated transcription have been investigated. A constitutive activated oestrogen receptor stimulated BKV promoter activity in HeLa cells. Mutations in either the ERE- or the AP-1-binding sites did not impair oestrogen receptor-induced activation of the BKV Dunlop promoter, while mutations in both binding motifs almost completely abolished oestrogen receptor-induced transcription. Simultaneous expression of large T- and small t-antigens strongly activated oestrogen receptor-mediated transcription. When expressed separately, only large T-antigen moderately stimulated oestrogen receptor-mediated transcription. The stimulatory effect of large T-antigen on the activity of the oestrogen receptor is probably indirect because no physical interaction between the two proteins was detected in a two-hybrid assay. Large T-antigen abrogated the synergistic effect on transcription between this nuclear receptor and the general co-activator CBP. The findings that the BKV early proteins amplify oestrogen receptor-mediated transcription may have important biological implications in individuals with raised oestrogen concentrations.

Antigens, Polyomavirus Transforming↗

PubMed1999

A dominant role for the Raf-MEK pathway in forskolin, 12-O-tetradecanoyl-phorbol acetate, and platelet-derived growth factor-induced CREB (cAMP-responsive element-binding protein) activation, uncoupled from serine 133 phosphorylation in NIH 3T3 cells.

In this study we describe that platelet-derived growth factor (PDGF), 12-O-tetradecanoyl-phorbol-acetate (TPA), and forskolin induced CREB (cAMP-responsive element-binding protein) Ser-133 phosphorylation with comparable magnitude and kinetics in NIH 3T3 cells. While forskolin was the most potent activator of CREB, TPA or PDGF modestly increased CREB activity. The role of protein kinase C, protein kinase A, and the Raf-MEK kinase pathway in the activation and Ser-133 phosphorylation of CREB by these three stimuli was investigated. We found that inhibition of the Raf-MEK kinase pathway efficiently blocks transcriptional activation of CREB by all three stimuli. This dominant involvement of Raf-MEK in CREB transcriptional activation seems to be uncoupled from CREB Ser-133 phosphorylation. We further demonstrate that although inhibition of Raf-MEK represses forskolin-induced CREB activation, forskolin by itself failed to activate ERK1/2 and Elk-1 mediated transcription. These results suggest that a basal level of Raf-MEK activity is necessary for both PDGF- and forskolin-induced CREB activation, independent of CREB Ser-133 phosphorylation.

3T3 Cells↗

PubMed1999

Distension-induced duodenal contractions vary with the phases of the canine interdigestive migrating motility complex.

BACKGROUND: The aim of the study was to evaluate whether the sensitivity for distension-induced contractions in the duodenum varied with the phases of the interdigestive migrating motility complex (MMC). METHODS: Four beagles (12-17 kg) with gastric fistulas were studied by means of perfused low-compliance manometry. A 2.5-cm-long balloon was placed in the third part of the duodenum. Side holes for pressure measurements were placed inside the balloon, 2cm proximal to the balloon, and in the antrum. Two-minute distensions with balloon pressures of 10 cmH2O were repeated in 10-min intervals during 17 MMCs. Twelve MMCs were also recorded without doing distensions. RESULTS: Balloon distension did not affect the duration of phase III and the MMC. In the beginning of phase I, distension did not significantly affect duodenal motility whereas distension induced contractile responses in late phase I, early phase II and late phase II (p < 0.05). The highest increase from the pre-distension to distension period was observed in early phase II (average 4 contractions min(-1)). However, considering the level of pre-distension contraction frequency, the fractional increase was by far the highest in late phase I. In phase III no difference was found between the periods before and during distension (15.8 +/- 0.7 versus 15.8 +/- 0.5 contractions min(-1)). The effect of balloon distension in phase IV was difficult to evaluate due to the inconsistent nature and sometimes rapidly decreasing activity in phase IV. Duodenal distension did not change frequency and amplitude of antral phase II contractions (p > 0.5). CONCLUSIONS: Duodenal balloon distension elicited ascending contractions in the canine duodenum in late phase I and in phase II. A refractory phase was demonstrated in the beginning of phase I while in phase III the frequency of contraction could not be increased further.

Animals↗

PubMed1998

Selective inhibitors of cytosolic or secretory phospholipase A2 block TNF-induced activation of transcription factor nuclear factor-kappa B and expression of ICAM-1.

TNF signaling mechanisms involved in activation of transcription factor NF-kappaB were studied in the human keratinocyte cell line HaCaT. We show that TNF-induced activation of NF-kappaB was inhibited by the well-known selective inhibitors of cytosolic phospholipase A2 (cPLA2): the trifluoromethyl ketone analogue of arachidonic acid (AACOCF3) and methyl arachidonyl fluorophosphate. The trifluoromethyl ketone analogue of eicosapentaenoic acid (EPACOCF3) also suppressed TNF-induced NF-kappaB activation and inhibited in vitro cPLA2 enzyme activity with a similar potency as AACOCF3. The arachidonyl methyl ketone analogue (AACOCH3) and the eicosapentanoyl analogue (EPACHOHCF3), which both failed to inhibit cPLA2 enzyme activity in vitro, had no effect on TNF-induced NF-kappaB activation. TNF-induced NF-kappaB activation was also strongly reduced in cells stimulated in the presence of the secretory PLA2 (sPLA2) inhibitors 12-epi-scalaradial and LY311727. Addition of excess arachidonic acid suppressed the inhibitory effect of 12-epi-scalaradial and LY311727. Moreover, both methyl arachidonyl fluorophosphate and 12-epi-scalaradial blocked TNF-mediated enhancement of expression of ICAM-1. Activation of NF-kappaB by IL-1beta was markedly less sensitive to both cPLA2 and sPLA2 inhibitors. The results indicate that both cPLA2 and sPLA2 may be involved in the TNF signal transduction pathway leading to nuclear translocation of NF-kappaB and to NF-kappaB-activated gene expression in HaCaT cells.

Animals↗

PubMed1998

[High-resolution computer tomography of the lungs].

High-resolution CT (HRCT) is a technique developed over the last decade. It optimises spatial resolution and provides details similar to those obtainable from gross pathologic specimens. HRCT of the lungs provides an accurate assessment of the pattern and distribution of many disease processes that in the conventional chest radiograph are occult or non-specific. HRCT is an established technique and the method of choice for evaluating a variety of pulmonary diseases. In this review we describe the modifications in CT technique that are instrumental in obtaining HRCT, and we also give examples of normal and pathologic findings in general.

Humans↗

PubMed1998

Synergistic increase in c-fos expression by simultaneous activation of the ras/raf/map kinase- and protein kinase A signaling pathways is mediated by the c-fos AP-1 and SRE sites.

Expression of the c-fos proto-oncogene is induced by numerous stimuli some of which are transmitted through the Ras/Raf/MAP kinase or the cAMP-dependent protein kinase (PKA) pathways. The effect of cell-specific interactions between these pathways on c-fos expression was investigated by exposing quiescent NIH3T3 cells to serum, forskolin, or a combination. Co-stimulation with serum and forskolin resulted in a more than additive increase in c-fos transcription. Synergistic increase in c-fos promoter activity was also observed in transient transfection studies after co-stimulation with serum plus forskolin or co-transfection with c-Raf and PKA expression plasmids. Analysis of the cAMP signaling pathway revealed that the synergy was neither due to an increase in PKA activity nor to Ser-133 phosphorylation/activation of CREB. The activation status of the MAP kinases ERK1 and ERK2 in co-treated cells was comparable to that in serum-treated cells. Co-stimulation with forskolin did not alter the phosphorylation state of Elk-1 compared to serum-induced phosphorylation of Elk-1. Deletion of c-fos promoter elements previously shown to be important for regulation of c-fos expression in response to mitogens indicates a role for SRE and FAP-1 elements.

3T3 Cells↗

PubMed1998

Mesocosm study of Mytilus edulis larvae and postlarvae, including the settlement phase, exposed to a gradient of tributyltin.

In a mesocosm study, the effects of a gradient of tributylin (TBT) (nominal TBT concentrations of 0.3, 2.3, 18.5, 146, and 1150 ng Sn liter-1) on Mytilus edulis larvae and postlarvae, including the settlement phase, were investigated over 15 days. Effects of TBT on mortality, growth measured as increase in shell length, shell dimensions, and settlement were evaluated. In general, mortality was high in all mesocosms including the control; during the first 24 h of the experiment, mortality was 63% at the highest TBT concentration (26% in the control). An LC50 (24 h) of 254 ng Sn liter-1 was estimated. The mortality rate of larvae/postlarvae increased 40% at 18.5 ng Sn liter-1 (0.41 day-1) compared with the control (0.30 day-1). For postlarvae, the growth rate decreased with increasing TBT concentration. The mean shell length at 2.3 ng Sn liter-1 was significantly reduced in comparison to the control on Day 15. Then EC10 (15 days) for shell growth was estimated to be 5.4 ng Sn liter-1. This is the lowest effect concentration ever reported in the literature. For postlarvae, shell dimensions in terms of shell length-shell width relations were affected by TBT at 1150 ng Sn liter-1, because the reduced growth led to the failure of adult mussels to secret a dissochonch shell. During the first days of exposure, the settlement monitored on polyethylene settling strips was stimulated by TBT, after which the settlement decreased due to the high mortality. Only a small portion of the population survived to the end of the test period. By comparison of the biotic conditions (in terms of larval abundance and particle concentration reflecting larval food) in the control mesocosm with those in the cove where the experiment was conducted, it was concluded that the mesocosm had successfully simulated the field conditions.

Animals↗

PubMed1998

Time course and pattern of pulmonary flow distribution following unilateral airway occlusion in sheep.

1. Unilateral bronchial occlusion causes ipsilateral hypoxic pulmonary vasoconstriction, which shifts blood flow towards the other lung. We studied the time course of flow diversion following acute bronchial occlusion, and the temporal effect of the latter on blood gases and vertical distribution of blood flow within the two lungs. 2. Serial infusion of radioactive or fluorescent microspheres were given to each of seven adult standing sheep before, during occlusion of the left mainstem bronchus for up to 6 min, and after release of occlusion. Pulmonary and systemic arterial pressures were recorded continuously and arterial and mixed venous blood gases were determined intermittently. Post-mortem, the lungs were inflated, dried and cut into slices. Relative blood flow at the time of infusion was expressed as the weight-normalized intensity of each tracer in each slice or lung divided by the weight-normalized intensity in the two lungs. 3. Within 30 s, 1 min and 2 min after onset of occlusion, flow in the occluded lung had decreased to 68-84% (range), 51-78% and 43-79% respectively, of the initial value. In the contralateral lung, flow increased by 10-24%, 14-37% and 23-39% respectively. The distribution of flow along the gravitational axis within each lung varied widely between animals, both before and during occlusion. The during-occlusion profiles in the occluded lung differed from those in the non-occluded lung. In either lung, during-occlusion profiles could not be predicted with certainty from the pre-occlusion profiles. Two minutes post-occlusion, inter- and intra-lung flow distribution were nearly the same as before occlusion. Arterial oxygen tension fell in the first minute of occlusion, but never below 7.5 kPa, and increased slowly thereafter. Arterial carbon dioxide tension increased slightly throughout the occlusion period. No appreciable changes in systemic or pulmonary artery pressure were observed. Post-occlusion, arterial oxygen tension was still sub-normal, while carbon dioxide tension continued to increase. 4. We conclude that acute unilateral bronchial occlusion diverts blood flow within 30 s towards the contralateral lung. This rapidly occurring flow diversion prevents the development of severe arterial hypoxaemia. The variable and largely unpredictable distribution of blood flow in the hyperfused non-occluded lung might explain some of the gas-exchange abnormalities observed in physiologically hyperfused lungs and in patients with one hyperfused lung.

Airway Obstruction↗

PubMed1997

A simple method for the preparation and purification of C1 complement cleaved beta 2-microglobulin from human serum.

A simple method is described for the preparation of proteolytically processed forms of beta 2-microglobulin suitable for structural and biological studies. PEG 6000 was added to the serum of healthy individuals to precipitate the C1 complement complex from C1 esterase inhibitor (C1-inh). After dissolving the precipitate containing the C1 complement in Tris-HCl buffer, pH 7.6, efficient conversion of added beta 2-microglobulin to desLys58 beta 2-microglobulin was observed. Addition of a specific carboxypeptidase B inhibitor (Plummers inhibitor) could partly prevent the deletion of Lys-58 from cleaved beta 2-microglobulin, whereby Lys58-cleaved beta 2-microglobulin was obtained. The proteolytically processed forms were subsequently purified by G-75 Sephadex gel filtration followed by chromatofocusing. A yield of 10-40% of proteolytically processed beta 2-microglobulin was obtained. Only one component was seen by SDS-PAGE stained with Coomassie Brilliant Blue.

Chromatography, Gel↗

PubMed1997

Mechanisms of transcriptional regulation of cellular genes by SV40 large T- and small T-antigens.

During the past decade a number of virus-encoded transcriptional trans-activators that regulate the expression of viral genes have been reported. These trans-activators may also affect the expression or activity of several cellular genes or gene products to create an optimal cellular environment that favors viral replication. Among the better-studied viral trans-activating proteins are the Simian virus 40 large T- and small t-antigens. During the last few years, mechanisms by which these two viral proteins influence cellular gene expression start to emerge. They are grouped provisionally and reflect the methods used to determine the effects of large T-antigen. Large T-antigen may influence cellular gene expression by: i. altering mRNA levels of cellular transcription factors; ii. interacting with and regulating the DNA-binding or transcriptional activity of specific transcription factors; iii. functionally substitution of eukaryotic transcription factors; iv. direct binding to DNA; or v. regulating components of signaling transduction pathways. Small t-ag seems to exert its effect mainly through inhibiting a cellular phosphatase, protein phosphatase 2A, thereby modulating components of signal transduction pathways and preventing dephosphorylation of several transcription factors. However, small t-ag may also control cellular gene expression by regulating mRNA levels of transcription factors or by interacting with other transcription factors.

Animals↗

PubMed1997

Localization of nonpancreatic secretory phospholipase A2 in normal and atherosclerotic arteries. Activity of the isolated enzyme on low-density lipoproteins.

Secretory nonpancreatic type II phospholipase A2 (snpPLA2) hydrolyzes fatty acids at the sn-2 position in phospholipids releasing free fatty acids (FFAs) and lysophospholipids. These products may act as intracellular second messengers or can be further metabolized into proinflammatory lipid mediators. The presence of snpPLA2 in extracellular fluids and serum during inflammation has suggested a role of the enzyme in this process. However, the presence of snpPLA2 in a variety of normal tissues suggests that snpPLA2 may also have physiological functions. Atherosclerosis appears to have an inflammatory component. Here we report on the snpPLA2 localization in normal and atherosclerotic lesions and on the properties of the isolated enzyme. A strong snpPLA2 immunoreactivity was observed in the arterial media that was colocalized with alpha-actin-positive vascular smooth muscle cells (SMCs) in both normal and atherosclerotic vessels. In aortic atherosclerotic lesions, snpPLA2 was observed colocalized with CD68-positive macrophages and HHF-35-positive SMCs and extracellularly in the lipid core. snpPLA2 was isolated from human normal arteries and from aorta with lesions. The enzyme was isolated by acid extraction of normal arterial tissues followed by immunoaffinity chromatography. The purified snpPLA2 had an expected molecular weight of 14 kD by polyacrylamide gel electrophoresis and appeared as a single band in immunoblotting. The enzymatic activity was followed by measuring release of fatty acids from phospholipid liposomes or LDL as substrates. The enzymatic activity was inhibited with two specific inhibitors for human snpPLA2: (1) monoclonal antibody 187 and (2) LY311727, a synthetic selective inhibitor. The mRNA for snpPLA2 was detected with reverse transcriptase polymerase chain reaction. These results indicate that snpPLA2 is present in human arteries and that it is able to hydrolyze phospholipids in LDL. The results support the hypothesis that snpPLA2 can release proinflammatory lipids at places of LDL deposition in the arterial wall.

Adult↗

PubMed1996

Binding of human phospholipase A2 type II to proteoglycans. Differential effect of glycosaminoglycans on enzyme activity.

Phospholipase A2 acting on low density lipoproteins in the extracellular arterial intima may form proinflammatory lipid mediators. Human nonpancreatic secretory phospholipase A2 has three regions that may associate with sulfated glycosaminoglycans. The apoB-100 molecule in low density lipoproteins also has glycosaminoglycan binding regions that could mediate its retention in the arterial intima. Here we report that human nonpancreatic phospholipase A2 isolated from a transfected cell line binds to glycosaminoglycans secreted by cultured human arterial smooth muscle cells. A gel mobility shift assay showed that the affinity of phospholipase A2 for glycosaminoglycans from a heparan sulfate/chondroitin sulfate proteoglycan was higher than for chondroitin sulfate glycosaminoglycans from a larger versican-like proteoglycan. Affinity chromatography confirmed these results. All glycosaminoglycans tested, at concentrations up to 100 microM, increased the activity of phospholipase A2 toward phosphatidylcholine liposomes. Above this concentration, heparan sulfate and heparin inhibited the enzyme. Heparin and chondroitin 6-sulfate increased phospholipase A2 activity on low density lipoproteins up to 4-fold at 100 microM, whereas heparan sulfate had no effect. The results indicate that human nonpancreatic secretory phospholipase A2 interacts with proteoglycans via their glycosaminoglycan moiety and that the enzyme activity may be modulated by the association of the enzyme and its substrate to the sulfated polysaccharides.

Chondroitin Sulfates↗

PubMed1996

PCR and sequencing from a single pollen grain.

In order to eliminate the laborious step of DNA extraction preceding all studies within the field of plant molecular biology we attempted to do PCR amplifications directly on pollen grains. Successful PCR amplification was obtained in reactions including a single pollen grain from Hordeum vulgare or Secale strictum. Both the plastid gene encoding ribulose-1,5-biphosphate carboxylase/oxygenase (rbcL) and the nuclear-encoded internal transcribed spacer regions (ITS) and the 5.8S rDNA region were amplified and sequenced to verify PCR amplification.

Hordeum↗

PubMed1996

Single lung transplantation for chronic obstructive pulmonary disease: pulmonary function and impact of bronchiolitis obliterans syndrome.

Chronic obstructive pulmonary disease (COPD) is now the most common indication for single lung transplantation. In long-term follow-up, obliterative bronchiolitis is a major problem. The aim of the present study was to perform a long-term follow-up of the pulmonary function and to examine the effect of development of bronchiolitis obliterans syndrome (BOS). Nineteen patients with end-stage COPD underwent single lung transplantation and were followed regularly with pulmonary function tests, and ventilation and perfusion scintigraphy (mean observation time 29 months). They were divided into two categories, with and without BOS, using the definition recommended by the International Society for Heart and Lung Transplantation working group. A mixed model analysis of variance with BOS as co-variate was used to evaluate its effect on pulmonary function. Spirometry, lung transfer factor for carbon monoxide (TLCO), arterial blood gases and 6-min walk test improved significantly (P < 0.001) from before transplantation to 3 months after transplantation. Nine patients developed BOS. Implied by the definition of the syndrome, forced expiratory volume in 1 s (FEV1) was significantly (P < 0.001) lower for patients with BOS while there was no significant effect of BOS category on TLCO corrected for alveolar volume (VA) or perfusion to transplant. Patients without BOS maintained their pulmonary function, and ventilation and perfusion to transplant for more than 3 yr after transplantation. The present results suggest that decreasing FEV1 accompanied by an unchanged TLCO/VA and pulmonary perfusion support the diagnosis of BOS after single lung transplantation for COPD.

Adult↗

PubMed1996

When to suspect and how to diagnose pulmonary lymphangioleiomyomatosis.

The objective of this study was to present clinical and radiological data of eight women with histologically proven lymphangioleiomyomatosis (LAM) diagnosed between 1984 and 1994, and to suggest a diagnostic strategy when LAM is suspected. A review of case reports, including results of biopsies, lung function and radiological procedures was undertaken. The mean age of the women at start of symptoms was 36 years, and the mean age at time of diagnosis 42 years. The most frequent presenting complaint was dyspnea, either in conjunction with pneumothorax (3), chylothorax (2) or on exertion (2). All patients had airflow limitation and markedly reduced gas transfer. Five patients had 16 episodes of pneumothorax. In seven patients multiple cysts were observed on the surface of the lung during thoracotomy while computerized tomography (CT) scans revealed numerous cysts evenly distributed throughout the lung parenchyma. The procedures that confirmed the diagnosis included transbronchial lung biopsy (4), open lung biopsy (2), thoracoscopy (1), thoracotomy (3) and autopsy (1). Three specimens had to be revised before the histological diagnosis was confirmed. It was concluded that the important clues to a diagnosis of LAM are recurrent episodes of pneumothoraces in fertile women, progressive air-flow limitation, markedly reduced gas transfer and characteristic findings on thoracic CT scans. A specific request to the pathologist to stain lung tissue specimens for smooth muscle cells is mandatory.

Adult↗

PubMed1996

Mechanical coupling between the hemithoraces in humans.

Unilateral airway occlusion permits measurement of single lung function. Since the results suggest that the occluded lung influences the mobility of the contralateral hemithorax, we wanted to test this hypothesis. In eight healthy subjects, we measured, using computed tomography, lung parenchymal area and inner rib cage parasagittal and transverse diameters at three different levels and at maximal inspiration and expiration. These measurements were made without and during balloon occlusion of one mainstem bronchus at residual volume (RV) and at total lung capacity (TLC). Unilateral occlusion at RV reduced the ipsilateral diameters in maximal inspiration, but the increase during inspiration was still 39-50% of that without occlusion. The inspiratory increase in contralateral diameters was reduced to 64-80% of the increase without occlusion. Occlusion at TLC reduced the expiratory decrease in ipsilateral diameters to 37-57% of that without occlusion. The expiratory decrease on the contralateral side was reduced to 56-70% of that without occlusion. Due to accompanying mediastinal shifts the parenchymal areas of the occluded lung barely changed. In contrast, the contralateral area was 86-97% of that without occlusion. We conclude that the movement of the two hemithoraces are, at least partially, interdependent. The occluded hemithorax prevents full expansion/compression of the nonoccluded contralateral side, whilst its own mobility appears to be increased by the presence of this nonoccluded side. Potential negative effects are outweighed by the physiological benefit of the coupling, as this mechanism could secure ventilation to a chest half without own movement.

Adult↗

PubMed1996

Acid-base status and cardiovascular function in mink (Mustela vison) anaesthetized with ketamine/midazolam.

Heart rate, arterial blood pressure and blood acid-base status were determined in 18 adult female mink (mean (+/- SEM) body weight 1052 +/- 34 g) during long-term anaesthesia with either controlled ventilation (n=12) or spontaneous respiration (n=6). Surgical anaesthesia was induced by intramuscular injection of ketamine hydrochloride (Ketaminol Vet, 40.0 +/- 1.7 mg/kg) and midazolam hydrochloride (Dormicum 2.8 +/- 0.1 mg/kg) and maintained for at least 5 h by continuous intravenous infusion of this drug combination in 0.9% saline. For all animals, the mean rates of infusion of ketamine and midazolam were 48.4 +/- 1.6 and 1.61 +/- 0.12 mg/h, respectively. Following continuous infusion of the anaesthetics in isotonic saline, at a rate of 20 ml/h, a moderate 'dilution acidosis' developed, which could be corrected by replacement of part of the saline with sodium bicarbonate to a final concentration of approximately 25 mmol NaHCO3 per litre. However, when the animals were allowed to breathe spontaneously, an increase in heart rate and a combined respiratory and metabolic acidosis occurred, due to severe respiratory depression. Apart from these effects and a few cases of increased salivation, no adverse effects over time were observed on the arterial blood acid-base status and cardiovascular function of the animals during ketamine/midazolam anaesthesia. It is concluded that the procedure described for long-term anaesthesia in mink is convenient and safe for acute physiological experiments in this species, provided normal body temperature and pulmonary gas exchange is sufficiently maintained. Thus, the need for an adequately controlled artificial ventilation is strongly emphasized. Finally, a proposal for the composition of an intravenous solution, containing ketamine and midazolam hydrochloride, and sodium bicarbonate in saline, suitable for long-term anaesthesia in adult mink is presented.

Acid-Base Equilibrium↗