Sex-specific reference ranges of cerebroplacental and umbilicocerebral ratios: longitudinal study.

OBJECTIVES: Observational studies have shown that low cerebroplacental ratio (CPR) values predict an increased risk of adverse perinatal outcome. The inverse ratio, i.e. the umbilicocerebral ratio (UCR), has been suggested to be a better predictor as it rises with increasing degree of fetal compromise. However, longitudinal reference ranges for UCR have not been established, and whether gestational-age-dependent changes in CPR or UCR differ between male and female fetuses has not been studied. Thus, the aims of this study were to investigate sex-specific, gestational-age-associated serial changes in CPR and UCR during the second half of pregnancy and to establish longitudinal reference ranges. METHODS: This was a secondary analysis of prospectively collected data from a dual-center longitudinal observational cohort study of low-risk singleton pregnancies. Doppler blood-flow velocity waveforms were obtained serially from the umbilical artery (UA) and fetal middle cerebral artery (MCA) from 19-41 weeks' gestation, and pulsatility indices (PIs) were determined. CPR and UCR were calculated as the ratios MCA-PI/UA-PI and UA-PI/MCA-PI, respectively. The course and outcome of pregnancies were recorded, and the sex of the fetus was determined after delivery. Reference intervals for CPR and UCR were constructed using multilevel modeling, and gestational-age-specific Z-scores in male and female fetuses were compared. RESULTS: Of a total of 299 pregnancies enrolled, 284 (148 male and 136 female fetuses) were included in the final analysis, and 979 paired measurements of UA-PI and MCA-PI were used to construct sex-specific longitudinal reference intervals. The relationship of both CPR and UCR with gestational age was U-shaped, but in opposite directions. There was a small but significant difference in Z-scores of CPR and UCR between male and female fetuses throughout the second half of pregnancy (P = 0.007). CONCLUSIONS: We have established longitudinal reference ranges for CPR and UCR suitable for serial monitoring, with the possibility of refining assessment by using fetal sex-specific ranges and conditioning by a previous measurement. The clinical significance of such refinements needs further evaluation. © 2019 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of the International Society of Ultrasound in Obstetrics and Gynecology.

Prediction of adverse perinatal outcome of small-for-gestational-age pregnancy using size centiles and conditional growth centiles.

OBJECTIVE: To test whether adding conditional growth centiles to size centiles of estimated fetal weight (EFW) improves prediction of adverse perinatal outcome in pregnancies with or at risk of having a small-for-gestational-age (SGA) fetus. METHODS: This prospective longitudinal study included pregnant women at risk of or diagnosed with an SGA (≤ 5(th) centile) fetus. They underwent serial ultrasound measurements and the final two were included in the analyses for this study. The EFW was categorized into normal (> 5(th) or 10(th) centile) and abnormal (≤ 5(th) or 10(th) centile) for size and conditional growth before entering the variables into log-binomial regression analyses. Adverse outcomes were delivery < 37 weeks, operative delivery due to fetal distress, 5-min Apgar score < 7, newborn hypoglycemia (glucose < 2.0 mmol/L), admission to the neonatal intensive care unit and perinatal mortality. A combined outcome variable ('any adverse outcome') included one or more adverse outcomes. RESULTS: Complete biometric data were obtained for 211 women. Conditional growth and size centiles contributed independently to the prediction of adverse outcome. Combining conditional growth and size centiles significantly improved the prediction of outcomes compared with size centiles alone (e.g. for 5(th) centile cut-off for any adverse outcome, P = 0.023, log-likelihood test). Using a 5(th) centile threshold, for any adverse outcome, the specificity of 78% (95% CI, 70-84%) using size centile as a predictor was improved to 94% (95% CI, 89-97%) when conditional growth centile was added to the model, whereas the sensitivity was not significantly changed (60% (95% CI, 49-69%) vs 39% (95% CI, 30-50%)). CONCLUSIONS: Size centiles and conditional growth centiles contribute independently to the prediction of adverse perinatal outcome, and their combination further improves the prediction model. The results support an increased use of conditional growth centiles in the monitoring of fetuses at risk. Copyright © 2015 ISUOG. Published by John Wiley & Sons Ltd.

Extracorporeal membrane oxygenation support for 59 days without changing the ECMO circuit: a case of Legionella pneumonia.

We report the successful use of veno-venous extracorporeal membrane oxygenation (ECMO) in a 53-year-old patient with Legionella pneumonia and acute respiratory distress syndrome (ARDS) with severe barotraumas. The patient was supported for 59 days without any changes in the ECMO circuit. This is probably the longest support ever reported using the same oxygenator.

Is it possible to predict outcome in pulmonary ECMO? Analysis of pre-operative risk factors.

Serious pulmonary failure may be treated with extracorporeal membrane oxygenation (ECMO) when other treatment has failed. The aim of this study was to analyze pre-operative risk factors of early mortality in patients who underwent either veno-arterial (VA) ECMO or veno-venous (VV) ECMO for pulmonary failure. We studied a total of 26 risk factors in 72 patients with severe pulmonary insufficiency treated with ECMO. All consecutive cases treated at our institution between Sept 1990 and Aug 2007 were included. Univariate analysis and multiple logistic regression analysis were performed on 26 risk factors. The end point was early mortality (any death within 30 days of ECMO treatment). Thirty-six (50%) of the patients died within 30 days of treatment. Age, gender, body mass index(BMI)(adults), cause of pulmonary failure, pre-ECMO treatment with nitric oxide(NO), intra-aortic balloon pump(IABP), and type of ventilation did not significantly influence early mortality. Neither pre-operative blood gas results, oxygenation index or pre-operative PaO(2)/FiO(2) ratio, nor mean ventilator days prior to ECMO gave any indications on early mortality. Liver function did not predict early mortality, but pre-ECMO serum creatinine levels were significantly lower in patients who survived. Treatment with ECMO in patients with severe pulmonary failure may save lives. It is, however, difficult to predict outcome when initiating ECMO. In this analysis, only pre-operative serum creatinine levels correlated with survival. None of the other parameters, including those which were used to select patients for ECMO treatment, could significantly predict the outcome.

Transportation of critically ill patients on extracorporeal membrane oxygenation.

Serious pulmonary and cardiac failure may be treated with extracorporeal membrane oxygenation (ECMO) when conventional treatment fails. In some severely ill patients, it may be necessary to initiate ECMO at the local hospital and, thereafter, transport the patient back to the ECMO center. The aim of this study was to evaluate our experiences with transportation of patients on ECMO. From Oct 1992 to Jan 2008 23, patients were transported on ECMO from local hospitals to Rikshospitalet. The study included seventeen patients with pulmonary failure and four patients with cardiac failure. All age groups were represented. Aircraft were used in 17 cases, ground vehicles in six. The times from decision until ECMO was established, the time from ECMO to departure from the local hospital and the transportation time were registered. All transportations were uneventful. After 10.3 +/-6.7 days, six patients died on ECMO and another patient died within 30 days. Mean ECMO time for those who died was 13.3 +/- 9.6 vs. 8.5 +/- 4.7 days for survivors, p=0.34. Seventeen patients were able to be successfully weaned from ECMO. Thirty day survival was 67%. The mean age for survivors was 15.3+/-18.3 (range 0-54.6) vs. 23.6 +/- 20.3 years (range 0-55.9) in fatal cases, p=0.41. The time from referral to initiating ECMO was a mean of 7.32 +/- 2.3 (3.0-12.0) hours for survivors vs. 7.88 +/- 3.0 (3.50-13.40) hours for non- survivors, p=0.76. The time from initiating ECMO to departure was 5.1 +/- 6.5 (0.58-23.75) hours in survivors vs. 9.1 +/- 6.8 (0.55-18.45) hours in non-survivors, p=0.18. Time from departure to arrival at Rikshospitalet was a mean of 3.2 (0.50-5.10) hours for survivors versus 2.5 (0.5-4.40) for non-survivors, p=0.41. This study shows that ECMO can be successfully established at local hospitals, using an experienced team, and that transportation of patients on ECMO can be performed safely and without technical difficulties. Survival for this group of patients did not differ from patients treated at the ECMO center.

Evaluation of oxygenators and centrifugal pumps for long-term pediatric extracorporeal membrane oxygenation.

OBJECTIVE: Two extracorporeal membrane oxygenation (ECMO) circuits for children under 10 kg were evaluated and compared for plasma leakage, hemolysis, blood transfusions, and durability. METHODS: Group A (n=20) was supported by ECMO circuits with the Minimax oxygenator and the Biomedicus centrifugal pump. Group B (n=10) was supported by ECMO circuits with the Lilliput 2 ECMO oxygenator and the Rotaflow centrifugal pump. RESULTS: ECMO circuit durability, as measured by oxygenator lifespan, was significantly better in Group B than in Group A (p = 0.04). There was significantly lower hemolysis, measured by plasma free hemoglobin, in Group B (p = 0.019), and patients in Group B had significantly less need for antithrombin III transfusion (p = 0.004). No plasma leakage was observed in Group B oxygenators, but plasma leakage was observed in all Group A oxygenators. CONCLUSION: The combination of a Rotaflow centrifugal pump and Lilliput 2 ECMO oxygenator in pediatric ECMO circuits improved durability and reduced circuit-induced hemolysis. This improvement may be due to the low priming volume, the oxygenator's plasma leakage resistance, the suspended rotor of the centrifugal pump, or a combination of these factors.

Detection of infrasound and linear acceleration in fishes.

Fishes have an acute sensitivity to extremely low-frequency linear acceleration, or infrasound, even down to below 1 Hz. The otolith organs are the sensory system responsible for this ability. The hydrodynamic noise generated by swimming fishes is mainly in the infrasound range, and may be important in courtship and prey predator interactions. Intense infrasound has a deterring effect on some species, and has a potential in acoustic barriers. We hypothesize that the pattern of ambient infrasound in the oceans may be used for orientation in migratory fishes, and that pelagic fishes may detect changes in the surface wave pattern associated with altered water depth and distant land formations. We suggest that the acute sensitivity to linear acceleration could be used for inertial guidance, and to detect the relative velocity of layered ocean currents. Sensitivity to infrasound may be a widespread ability among aquatic organisms, and has also been reported in cephalopods and crustaceans.

Comparison of three oxygenator-coated and one total-circuit-coated extracorporeal devices.

The present study was designed to compare the biocompatibility of three cardiopulmonary bypass setups with different surface coatings, and to determine if coating of the whole circuit with one of the coatings was more beneficial than coating of the oxygenator only. Extracorporeal devices entirely coated with synthetic polymers (Avecor, n = 6) were compared to oxygenators coated with synthetic polymers (Avecor, n = 6), end-point, covalently attached heparin (CBAS, n = 6) or absorbed heparin (Duraflo 2, n = 6) in an in vitro model of a heart lung machine. The circuits were primed with fresh human whole blood and Ringer's acetate and recirculated at 4 l/min at 30 degrees C for 2 h. Test samples were obtained at regular intervals and analysed for myeloperoxidase (MPO), platelet counts, beta-thromboglobulin, heparin, prothrombin fragment 1+2, plasmin anti-plasmin complexes, and complement activation products. The mean MPO concentrations increased in the Avecor-coated oxygenator group (AV) from 247 at the start to 671 microg/l at the termination of the experiments, in the Avecor-coated total circuit group (AV-T) from 116 to 288 microg/l, in the Duraflo 2 coated oxygenator group (DU) from 160 to 332 microg/l, and in the CBAS-coated oxygenator (CA) group from 172 to 311 microg/l. The MPO concentrations increased significantly in all groups (p < 0.03). The increase in group A was significantly higher than in the other three groups (p = 0.007). The mean platelet counts decreased in the Avecor-coated total circuit group from 117 at start to 99 x 10(9)/l at termination of the experiments, in the Avecor-coated oxygenator group from 119 to 103 x 10(9)/l, in the Duraflo 2 group from 96 to 86 x 10(9)/l, and in the CBAS group from 132 to 123 x 10(9)/l. The platelet counts decreased significantly in all groups (p < 0.01), but the intergroup differences were not significant (p = 0.15). The mean beta-thromboglobulin concentrations increased in the Avecor-coated total circuit group from 193 at the start to 754 ng/ml at the termination of the experiments, in the Avecor-coated oxygenator group from 474 to 1654 ng/l, in the Duraflo 2 group from 496 to 1280 ng/l, and in the CBAS group from 418 to 747 ng/l. The beta-thromboglobulin increase was significant in each group (p < 0.01), but not between the groups (p = 0.49). The mean heparin concentrations in the Duraflo 2 group increased from 2460 at the start to 2897 IU/l at termination of the experiments, in the CBAS group from 2468 to 2518 IU/l. In the Avecor-coated oxygenator group heparin concentrations decreased from 2010 to 1968 IU/l, and in the Avecor-coated total circuit group from 2002 to 1927 IU/l. The differences in heparin concentrations were significant between the Duraflo 2 group and the other groups (p < 0.05). The mean prothrombin fragment 1+2 concentrations increased in the CBAS group from 0.4 at the start to 2.1 nmol/l at the end of the experiments, in the Avecor-coated oxygenator group from 0.4 to 0.6 nmol/l, in the Avecor-coated total circuit group from 0.3 to 0.4 nmol/l, and in the Duraflo 2 group from 1.2 to 1.3 nmol/l. The prothrombin fragment 1+2 increase was significant in all groups (p < 0.05), but there were no significant intergroup differences (p = 0.54). There were no significant differences at the termination of the experiments among the four groups regarding complement activation as measured by C3 activation products and the terminal complement complex. In the present in vitro model of a heart-lung machine, none of the three specific setups with different coatings was superior with regard to all test parameters. The CBAS group generated the highest levels of prothrombin fragment 1+2 formation, but least complement activation. The increasing plasma heparin concentrations in the Duraflo 2 group indicated more unstable heparin bonding. The Avecor-coated total circuit group were superior to the Avecor-coated oxygenator group regarding plasma concentrations of MPO, but not compa

In vitro evaluation of new surface coatings for extracorporeal circulation.

Cardiopulmonary bypass (CPB) exposes blood to large, foreign surfaces. This exposure may activate the cellular and humoral inflammatory systems, resulting in inflammatory reactions and organ dysfunction. Coating the inner surfaces of the bypass circuit may help alleviate these side-effects. The objective of this study was to determine the influence of two new surface treatments on blood cell and complement activation. Oxygenator and tubing sets coated with synthetic polymers (n = 7) or heparin (n = 7) were compared to uncoated sets (n = 7) in an in vitro model of CPB. The circuits were run at 4 l/min and recirculated for 120 min. The inflammatory response was assessed at regular intervals by platelet counts, and activation of complement, leucocytes and platelets. We found that the median platelet counts decreased from 127 to 122 x 10(9)/l (not significant, NS) in the synthetic polymer sets, from 96 to 88 x 10(9)/l (NS) in the heparin-coated sets, and from 93 to 54 x 10(9)/l (p < 0.01) in the uncoated sets after 2 h of recirculation. There were significant differences in platelet counts between the coated sets and the uncoated set at end of experiments (p < 0.05). Beta-thromboglobulin (BTG) concentrations increased in the synthetic polymer sets from 166 to 352 ng/ml (p < 0.01), in the heparin coated sets from 336 to 1168 ng/ml (p < 0.01), and in the uncoated sets from 301 to 3149 ng/ml (p < 0.01) after 2 h of recirculation. The differences in BTG at termination of the experiments were significant among all three sets (p < 0.05). Myeloperoxidase (MPO) concentrations in the synthetic polymer sets increased from 63 to 86 micrograms/l (p < 0.01), in the heparin-coated sets from 90 to 208 micrograms/l (p < 0.01), and in the uncoated sets from 122 to 513 micrograms/l (p < 0.01) after 2 h of recirculation. The differences in MPO at termination of the experiments were significant among all three groups (p < 0.01). There were no significant differences at termination of the experiments among the three sets regarding complement activation as measured by C3 activation products and the terminal complement complex. We conclude that in the current in vitro model of a CPB circuit, the synthetic polymer coating and the heparin coating caused significantly less platelet loss and granulocyte and platelet activation than the uncoated surface (p < 0.05). The synthetic polymer coating caused significantly less granulocyte and platelet activation than the heparin coating (p < 0.05). There was moderate complement activation within each group, but no significant differences among the three groups.

Extracorporeal membrane oxygenation using a centrifugal pump and a servo regulator to prevent negative inlet pressure.

BACKGROUND: We studied whether negative inlet pressure created by a centrifugal pump during extracorporeal membrane oxygenation damages blood. METHODS: Fresh, whole human blood and primer were circulated through a test circuit, applying an inlet pressure of 0, -50, or -100 mm Hg. Thereafter, hemolysis and kidney function were compared between 6 patients treated before and 14 patients treated after inclusion in our setup of extracorporeal membrane oxygenation with a servo inlet pressure regulator. RESULTS: In vitro, negative inlet pressure caused substantial hemolysis, leukocyte and platelet destruction, and complement activation. Maximal plasma free hemoglobin concentrations were 199 mg/100 mL before use of the servo inlet pressure regulator and 40 mg/100 mL afterward (p = 0.06), and serum creatinine peaked at 330 and 115 mumol/L, respectively (p = 0.03). The minimal 24-hour diuresis normalized for weight was 4.8 mL/kg before use of the servo inlet pressure regulator and 45.6 mL/kg afterward (p = 0.03). Three of 5 evaluable patients before use of the servo inlet pressure regulator and 1 of 14 patients after inclusion in this setup experienced anuria (p = 0.04). CONCLUSIONS: There were strong indications that reduction of negative pump inlet pressure with the servo regulator prevented hemolysis and kidney damage.

Effects on complement, granulocytes and platelets of a leukocyte-depletion filter during in vitro extracorporeal circulation.

In an in vitro study, extracorporeal circuits equipped with either a leukocyte-depleting filter (n = 5) or a standard arterial-line filter (n = 5) were perfused for 120 minutes with fresh human whole blood. Leukocyte activation, leukocyte and platelet counts and complement activation were studied. Significant reduction of leukocyte and platelet counts and significant activation of leukocytes and of platelets were found in both groups, but without significant intergroup difference for any parameter after 120 minutes of perfusion. The leukocyte-depleting filters, however, were somewhat more effective in removing leukocytes during the initial 30 minutes of circulation.

Efflux of gamma-aminobutyric acid caused by changes in ion concentrations and cell swelling simulating the effect of cerebral ischaemia.

The relationships among ischaemic GABA efflux from brain tissue and extracellular and intracellular concentrations of sodium, chloride and potassium ions were investigated by means of 1) transverse hippocampal slices from rat and 2) functional expression of a high affinity GABA transporter in Xenopus oocytes. Brain slices were incubated for 20 min in medium where extracellular sodium and chloride were substituted with impermeant ions. Isethionate (Iseth) substitution for chloride generated a 7-fold increase in GABA efflux. Choline (Chol) but not N-methyl-D-glucamine (NMDG) substitution for sodium likewise increased GABA efflux. Reducing the osmolarity of the medium by decreasing both sodium and chloride concentrations (Hyp) increased GABA efflux 3-fold. This release was blocked by mannitol (Man). Blocking sodium channels with 1 microM of tetrodotoxin (TTX) also increased the release 3-fold. Energy deprivation (ED) increased the GABA release 50-fold. ED/Iseth left the release unchanged, ED/Chol increased the GABA efflux by 23%, whereas ED/NMDG reduced the release by 41%. Adding mannitol did not block the ED-evoked release, whereas TTX reduced it by 52%. Release of preloaded [3H]-GABA from oocytes expressing the GAT-1 GABA transporter was then examined. Depolarisation by current injection or 100 mM extracellular K+ did not increase GABA release. Sodium chloride injection, however, caused membrane depolarisation and a 100-fold increased GABA efflux from the oocytes. This release was blocked when the osmolarity was increased extracellularly by adding mannitol. These results show that 1) TTX releases GABA from brain tissue but blocks release during ED, 2) the high affinity GABA carrier must be altered in order to reverse, 3) ischaemic GABA release is sodium independent, and is modulated by large cations, 4) mannitol blocks the reversal of high affinity carriers in oocytes, but the release from brain slices during ED is unaffected. Taken together, the results suggest that ischaemic release of GABA from brain tissue does not occur by means of reversed high affinity carriers alone, but rather that it is controlled by more complex mechanisms.

[From a registry to a clinical information system. Development of the Datacor system at the surgery department A, Rikshospitalet]. / Fra virksomhetsregistrering til klinisk informasjonssystem. Utvikling av Datacor-systemet ved Kirurgisk avdeling A, Rikshospitalet.

Based on a simple register for thoracic and cardiovascular operations a modulated system has been built up at Department of Surgery A. The register covers waiting list, a basic patient record, extensive operative data, the postoperative course and the final outcome. A local area network includes 36 microcomputers with approximately 75 users. Owing to lack of commercially available programs, local applications based on dBase have been developed. In this article we discuss our positive experiences from use of the local system with respect to administration, quality assurance and local research, its future place within a larger hospital system interconnected via a backbone, the need for better support and graphic user interface.

Complement activation during extracorporeal circulation. In vitro comparison of Duraflo II heparin-coated and uncoated oxygenator circuits.

The degree of complement activation during cardiopulmonary bypass is considered a valuable parameter of biocompatibility of the extracorporeal circuit. In an in vitro setting with a heart-lung machine primed with fresh whole blood and saline solution, the C3 activation products C3b, iC3b, and C3c and the terminal complement complex were measured in double-antibody enzyme immunosorbent assays. No differences were found between seven sets treated with Duraflo II heparin coating and seven uncoated sets after 2 hours of circulation. C3 activation products (expressed as median and 95% confidence intervals) increased from 4.5 AU (2.8 to 12.3 AU) to 16.5 AU (10.0 to 19.4 AU) in the uncoated sets (p = 0.02) and from 4.6 AU (2.2 to 5.8 AU) to 19.3 AU (3.5 to 27.1 AU) in the coated sets (p = 0.02). Terminal complement complex increased from 5.7 AU (2.7 to 11.3 AU) to 13.6 AU (8.2 to 17.8 AU) in the uncoated sets (p = 0.02) and from 7.9 AU (4.6 to 11.4 AU) to 17.3 AU (9.4 to 35.1 AU) in the coated sets (p = 0.02). A significant drop in thrombocyte levels was observed in both coated and uncoated sets. In a supplementary series, the sterilization process did not influence the results. Although Duraflo II heparin coating is considered highly effective in preventing coagulation, it did not prevent complement activation in the present in vitro study. We hypothesize that the mode by which the heparin molecule is bound to the surface may be essential to obtain effects on both coagulation and complement system.

Molecular cloning of a functional human thyrotropin-releasing hormone receptor.

A cDNA encoding the human thyrotropin-releasing hormone receptor (hTRH-R) was isolated from a human brain cDNA library. Screening of 1.2 million clones resulted in 2 candidates. The largest clone contained TRH-R homologous sequences starting in the third transmembrane domain and included a long 3' untranslated sequence. The smaller clone contained a potential start of the open reading frame, but was interrupted by an intron in the sixth transmembrane domain. The two clones had 497 bp of overlapping identical sequences and it was possible to assemble a complete cDNA thus restoring the assumed coding sequence. Electrophysiological studies of frog oocytes injected with in vitro transcribed mRNA showed TRH-specific inward currents, demonstrating that the reconstituted cDNA encoded a functional receptor. The predicted amino acid sequence of the hTRH-R protein showed high homology with the rat and mouse TRH-Rs with the exception of their C-terminal region. The human TRH-R gene seems to contain two introns.

Different-sized mRNAs from GH4C1 cells induce a TRH-dependent electrical response in Xenopus laevis oocytes.

Poly(A)+ RNA from the GH4C1 rat pituitary cell line elicited a thyrotropin releasing hormone response in Xenopus laevis oocytes which could be measured as a change in membrane current by the voltage-clamp method. Oocytes injected with Poly(A)+ RNA from GH12C1 cells which do not bind thyrotropin releasing hormone or with buffer solution alone did not show this response. Size fractionation of total poly(A)+ RNA by sucrose density-gradient centrifugation shows two response maximal representing various mRNA fractions larger than 18S. These results indicate the presence of thyrotropin releasing hormone receptor mRNA heterogeneity where the smallest mRNA is at least 2 kb.

Effect of Diamphidia toxin, a Bushman arrow poison, on ionic permeability in nucleated cells.

The effect of Diamphidia toxin, isolated from pupae of Diamphidia nigro-ornata, was tested on two different cell lines (GH4C1 cells and HL-60 cells) and on human lymphocytes. The toxin raised intracellular Ca2+ concentration, as assessed with quin 2, in a dose-related manner in all three cell types. The effect was abolished when extracellular Ca2+ was chelated by EGTA. Low concentrations of the toxin evoked a delayed as well as a smaller response. The response time was also temperature-dependent, with a Q10 of about 2. Low, but effective concentrations of the toxin did not affect cell membrane integrity, as tested with Trypan blue, and induced a seemingly physiological release of prolactin from the GH4C1 cells. Diamphidia toxin's effect on the membrane permeability of GH4C1 cells was further investigated with patch-clamp techniques. The toxin appeared to increase the conductance for all small ions without affecting the normal ionic channels present in these cells. We conclude that Diamphidia toxin has a general effect on the plasma membrane of different cell types and thereby increases, probably non-specifically, the permeability for small ions.