Immunomodulating treatment with low dose interleukin-4, interleukin-10 and interleukin-11 in psoriasis vulgaris.

Psoriasis is a chronic inflammatory skin disease affecting approximately 2-3 percent of the world population; it is characterised by hyperproliferation and hyperplasia of the superficial layers of the epidermis. Inappropriate signals released by the immune system determine an altered keratinocyte differentiation, resulting in the formation of desquamating, thickened, inflamed and erythematous plaques. The aim of this investigation was to study the pharmacological activity and safety of three low dose cytokines, Guna-Interleukin 4, Guna-Interleukin 10 and Guna-Interleukin 11 at the concentration of 10 fg/ml in patients affected by moderate to slight psoriasis vulgaris. The multicenter, double-blind, randomized, placebo-controlled clinical trial involved 48 patients who were enrolled and followed up according to a 8-month experimental project. All patients received, according to a cross-over model, either the experimental treatment or placebo, alternatively. Globally, in the 41 evaluated patients it was observed a PASI significant reduction (Friedman test: p=0.00960). The DLQI too decreased significantly in all subjects compared to baseline (Friedman test: p=0.00007). The safety of the treatment with three low dose cytokines administered simultaneously was proved; no adverse event was reported during the whole trial.

Uterine artery Doppler evaluation in twin pregnancies at 11 + 0 to 13 + 6 weeks of gestation.

OBJECTIVES: To compare uterine artery pulsatility index (PI) obtained at 11 + 0 to 13 + 6 weeks of gestation in singleton and twin pregnancies and to evaluate changes in PI values of twin pregnancies developing pre-eclampsia (PE) or small-for-gestational age (SGA) of either one or both fetuses. METHODS: Uterine artery PI was measured in 421 twin pregnancies (384 dichorionic and 37 monochorionic) and in 500 singleton pregnancies. The measured mean and lowest uterine artery PI values were converted to multiples of the expected normal median (MoM) after correction for maternal body mass index, ethnicity and gestational age. The median PI-MoM values of twins were compared with those of singleton pregnancies. In twin pregnancies, PI-MoM values were analyzed according to chorionicity, development of early-onset (< 34 weeks) or late-onset (≥ 34 weeks) PE and SGA of one or both twins. RESULTS: Uterine artery PI-MoM was significantly lower in twin compared with singleton pregnancies (mean K = 174.31, P < 0.0001, lowest K = 139.27, P < 0.0001). However, there were no significant differences in the uterine artery PI-MoM values between monochorionic and dichorionic twins. The uterine artery PI in twin pregnancies that developed early-onset PE (P < 0.001) and SGA of both twins (P < 0.05) was higher than the uterine artery PI in uncomplicated twin pregnancies, whereas no differences were found for late PE or SGA of one twin. CONCLUSIONS: First-trimester placental impedance to flow, as assessed by uterine artery Doppler examination, is reduced in twin pregnancies, with no differences related to chorionicity. The relative increase of uterine artery PI found in twin pregnancies that developed early PE and SGA of both twins suggests that first-trimester uterine artery assessment may be useful in identifying such complications.

An algorithm based on OmniView technology to reconstruct sagittal and coronal planes of the fetal brain from volume datasets acquired by three-dimensional ultrasound.

OBJECTIVE: To describe a novel algorithm, based on the new display technology 'OmniView', developed to visualize diagnostic sagittal and coronal planes of the fetal brain from volumes obtained by three-dimensional (3D) ultrasonography. METHODS: We developed an algorithm to image standard neurosonographic planes by drawing dissecting lines through the axial transventricular view of 3D volume datasets acquired transabdominally. The algorithm was tested on 106 normal fetuses at 18-24 weeks of gestation and the visualization rates of brain diagnostic planes were evaluated by two independent reviewers. The algorithm was also applied to nine cases with proven brain defects. RESULTS: The two reviewers, using the algorithm on normal fetuses, found satisfactory images with visualization rates ranging between 71.7% and 96.2% for sagittal planes and between 76.4% and 90.6% for coronal planes. The agreement rate between the two reviewers, as expressed by Cohen's kappa coefficient, was > 0.93 for sagittal planes and > 0.89 for coronal planes. All nine abnormal volumes were identified by a single observer from among a series including normal brains, and eight of these nine cases were diagnosed correctly. CONCLUSIONS: This novel algorithm can be used to visualize standard sagittal and coronal planes in the fetal brain. This approach may simplify the examination of the fetal brain and reduce dependency of success on operator skill.

First-trimester umbilical vein blood flow in pregnancies with low serum pregnancy-associated plasma protein-A levels: an early predictor of fetal growth restriction.

OBJECTIVE: To investigate umbilical vein blood flow (UVBF) during the first trimester in pregnancies with low serum pregnancy-associated plasma protein-A (PAPP-A) levels and to relate umbilical vein (UV) diameter, time-averaged maximum velocity (TAMXV) and UVBF values to the subsequent development of fetal intrauterine growth restriction (IUGR). METHODS: UVBF assessment was performed at 11 + 0 to 13 + 6 weeks' gestation in 102 singleton pregnancies with PAPP-A concentrations of < 0.3 multiples of the median. UV diameter, UV-TAMXV and UVBF were calculated and analyzed in relation to pregnancy outcome. RESULTS: Pregnancy outcomes were: 51 pregnancies with birth weight ≥ 10(th) centile (Group A), 30 pregnancies with birth weight < 10(th) centile with normal Doppler in the umbilical artery throughout gestation (Group B) and 21 pregnancies with birth weight < 10(th) centile and abnormal umbilical artery Doppler later in gestation (Group C). No differences were found in PAPP-A levels between groups. Group C fetuses exhibited significantly lower values of UV-TAMXV (z-score - 1.99 SDs, t = 8.527, P ≤ 0.0001) and UVBF (z-score - 0.97 SDs, t = 7.420, P ≤ 0.0001) in comparison with normal reference ranges, while no differences were found in Groups A or B. CONCLUSIONS: Decreased UV-TAMXV and UVBF at 11 + 0 to 13 + 6 weeks' gestation identify fetuses at risk of developing IUGR among pregnancies with low levels of PAPP-A.

Low cardiac output to the placenta: an early hemodynamic adaptive mechanism in intrauterine growth restriction.

OBJECTIVE: A low combined cardiac output (CCO) to the placenta (placenta/CCO fraction) has been reported in growth-restricted (IUGR) fetuses, but the temporal sequence of these modifications in relation to other changes in the fetal circulation is unknown. The aim of this study was to evaluate the placenta/CCO fraction in relation to other hemodynamic changes in fetuses at risk of developing IUGR. METHODS: We studied 340 singleton nulliparous pregnancies characterized at 20-24 weeks by abnormal uterine artery pulsatility index (PI) values (> 95(th) centile). At this gestational age we measured fetal biometry and Doppler waveforms from the umbilical artery (UA), middle cerebral artery (MCA), ductus venosus (DV), umbilical vein (UV) and outflow tracts of both ventricles. The diameters of the semilunar valves and UV were measured and CCO (left cardiac + right cardiac outputs) and UV blood flow were calculated. The placenta/CCO fraction was calculated as UV flow as a percentage of CCO. RESULTS: There were 283 pregnancies with birth weight >or= 10(th) centile and normal UA-PI throughout gestation (Group A), 34 with birth weight < 10(th) centile and normal UA-PI throughout gestation (Group B) and 23 with birth weight < 10(th) centile and abnormal UA-PI developing later in gestation (Group C). At 20-24 weeks there were no differences among the three groups in fetal biometric parameters, PI values from the UA, MCA and DV, and CCO. UV flow and placenta/CCO fraction were significantly lower in Group C compared with Group A (UV flow delta value = - 1.439, P < 0.0001; placenta/CCO fraction delta value = - 1.74, P < 0.0001) but not in Group B. CONCLUSIONS: Our data suggest that, in fetuses developing IUGR secondary to placental compromise, UV flow and placental/CCO fraction are already reduced by 20-24 weeks, and that this reduction occurs earlier than do modifications in fetal size and arterial and venous PI values.

Placental vascularization measured by three-dimensional power Doppler ultrasound at 11 to 13 + 6 weeks' gestation in normal and aneuploid fetuses.

OBJECTIVE: To establish the potential role of three-dimensional (3D) power Doppler evaluation of the placental circulation in aneuploidy screening at 11 to 13 + 6 weeks of gestation. METHODS: 3D power Doppler ultrasound examination of the placenta was performed in 25 pregnancies with fetuses with abnormal karyotype and in 100 control pregnancies at 11 to 13 + 6 weeks of gestation. Using the same pre-established settings for all cases, the vascularization index (VI), flow index (FI) and vascularization flow index (VFI) were calculated for the whole placenta. RESULTS: In the chromosomally normal group all the vascular indices increased significantly with advancing gestation between 11 and 13 + 6 weeks (VI: r = 0.482, P < 0.001; FI: r = 0.295, P = 0.0029; VFI, r = 0.484, P < 0.001). In the chromosomally abnormal group, the flow indices were not significantly different from normal in cases with trisomy 21 (13 cases), but they were significantly reduced compared with normal in cases with trisomies 13 and 18 (VI: t = 8.321, P < 0.0001: FI: t = 12.934, P < 0.0001; VFI: t = 7.608, P < 0.0001). CONCLUSIONS: 3D power Doppler evaluation of the placental circulation is not useful in screening for trisomy 21, and unlikely to further increase the already high detection rate for trisomies 13 and 18. However, we provide normal ranges of placental vascular indices between 11 + 0 and 13 + 6 weeks of gestation, which may be useful in future research on placental vascularity in certain at-risk pregnancies.

Connexin40 regulates renin production and blood pressure.

Renin secretion is regulated by coordinated signaling between the various cells of the juxtaglomerular apparatus. The renin-secreting cells (RSC), which play a major role in the control of blood pressure, are coupled to each other and to endothelial cells by Connexin40 (Cx40)-containing channels. In this study, we show that Cx40 knockout (Cx40-/-) mice, but not their heterozygous littermates, are hypertensive due to the increase in the number of RSC, renin biosynthesis, and plasma renin. Treatment with the angiotensin II receptor AT1 antagonist candesartan or the angiotensin II-converting enzyme inhibitor ramipril reduced the blood pressure of the Cx40-/- mice to the same levels seen in wild-type (WT) mice. The elevated blood pressure of the knockout mice was not affected by clipping one renal artery (2K1C, renin-dependent model of hypertension) or after a high salt diet. Under these conditions, however, Cx40-/- mice showed an altered production and release of renin. The renin mRNA ratio between the clipped and the non-clipped kidney was lower in the knockout than in the WT 2K1C mice. This indicates that the response to a change in blood pressure was altered. The RSC of the Cx40-/- mice did not have a compensatory increase in the levels of either Cx43 or Cx37. Our data show that renin secretion is dependent on Cx40 and suggest the Cx40-/- mice may be a genetic model of renin-dependent hypertension.

Accuracy of the ICD-9 codes for identifying TIA and stroke in an Italian automated database.

The object of this study was to evaluate the sensitivity and positive predictive value (PPV) of International Classification of Diseases, 9th revision (ICD-9) codes 430-438 in the Sistema Informativo Sanitario Regionale (SISR), an Italian health care automated database. We compared the SISR with a manual search of all cases of transient ischaemic attack (TIA) and stroke discharged from the Novara Hospital, NW Italy. Results were as follows: SISR list: 1017 patients; manual list 1005. Linked: 896; false negatives: 109; false positives: 121. Sensitivity of codes 430-438: 77% at the primary position only and 89% at either the primary or secondary position; PPV: 93% and 88%. Sensitivity and PPV for specific codes vs. each subcategory (sensitivity at the primary position only/any position; PPV at the primary position only/any position): for 430, subarachnoid haemorrhage (33/35%; 46/43%); for 431, cerebral haemorrhage (57/59%; 77/75%); for 434, cerebral infarction (35/37%; 90/87%); for 436, stroke of unknown type (29/29%; 19/16%); and for 435, TIA (75/82%; 80/78%). The SISR database has a high PPV; sensitivity is high for TIA, but low for specific stroke ICD codes.

Abnormal cardiac function in fetuses with increased nuchal translucency.

OBJECTIVE: To evaluate cardiac function in structurally and chromosomally normal fetuses with increased nuchal translucency (NT). METHODS: Forty-two structurally and chromosomally normal fetuses with increased NT at 11-14 weeks of gestation underwent fetal echocardiographic examination at 20-23 weeks. Fifty fetuses with normal NT values were considered as controls. Pulmonary and aortic peak velocity and time to peak velocities were measured as indices of ventricular systolic function. The ratios between the E-wave and A-wave (E/A) and the ratios between the E-wave and time velocity integral (E/TVI) at the level of both atrioventricular valves were evaluated as indices of ventricular diastolic function. RESULTS: In fetuses with increased NT the E/A ratios were significantly decreased when compared to control fetuses at the level of both the mitral (0.52 +/- 0.09 vs. 0.60 +/- 0.10, P = 0.0002) and tricuspid (0.51 +/- 0.09 vs. 0.61 +/- 0.09, P < 0.0001) valves. Similar results were found for the E/TVI ratios (mitral valve 4.79 +/- 1.03 vs. 5.63 +/- 1.23, P = 0.0007 and tricuspid valve 4.40 +/- 0.88 vs. 5.19 +/- 0.82, P < 0.0001). No significant relationship was found between the degree of NT and the abnormalities in Doppler indices. There were no significant differences in Doppler systolic indices. CONCLUSION: Structurally and chromosomally normal fetuses with increased NT have low E/A and E/TVI ratios at 20-23 weeks of gestation. These findings might indicate cardiac diastolic dysfunction.

Angiotensin II promotes selective uptake of high density lipoprotein cholesterol esters in bovine adrenal glomerulosa and human adrenocortical carcinoma cells through induction of scavenger receptor class B type I.

Angiotensin II is one of the main physiological regulators of aldosterone biosynthesis in the zona glomerulosa of the adrenal cortex. The hormone stimulates intracellular cholesterol mobilization to the mitochondrion for steroid biosynthesis. Here we have examined whether angiotensin II also modulates exogenous lipoprotein cholesterol ester supply to the steroidogenic machinery and whether this control is exerted on the selective transport of high density lipoprotein-derived cholesterol ester to intracellular lipid droplets through the scavenger receptor class B type I. In bovine adrenal glomerulosa and human NCI H295R adrenocortical carcinoma cells, high density lipoprotein stimulated steroid production. Angiotensin II pretreatment for 24 h potentiated this response. Fluorescence microscopy of cellular uptake of reconstituted high density lipoprotein containing a fluorescent cholesterol ester revealed an initial, time-dependent narrow labeling of the cell membrane followed by an intense accumulation of the fluorescent cholesterol ester within lipid droplets. At all time points, labeling was more pronounced in cells that had been treated for 24 h with angiotensin II. Fluorescence incorporation into cells was prevented by a monoclonal antibody directed against apolipoprotein A-I. Upon quantitative fluorometric determination, cholesterol ester uptake in angiotensin II-treated bovine cells was increased to 175 +/- 15% of controls after 2 h and to 260 +/- 10% after 4 h of exposure to fluorescent high density lipoprotein. The amount of scavenger receptor class B type I protein detected in cells treated with angiotensin II for 24 h reached 203 +/- 12% of that measured in control cells (n = 3, P < 0.01). In contrast, low density lipoprotein receptors were only minimally affected by angiotensin II treatment. This increase in scavenger receptor class B type I protein was associated with a 3-fold induction of scavenger receptor class B type I mRNA, which could be prevented by actinomycin D but not by cycloheximide. Similar results were obtained in the human adenocarcinoma cell line H295R. These observations show that angiotensin II regulates the scavenger receptor class B type I-mediated selective transport of lipoprotein cholesterol ester across the cell membrane as a major source of precursor for mineralocorticoid biosynthesis in both human and bovine adrenal cells.

Decreased expression of steroidogenic acute regulatory protein: a novel mechanism participating in the leptin-induced inhibition of glucocorticoid biosynthesis.

The adipocyte-derived hormone leptin is a central modulator of food intake, metabolism and neuroendocrine functions. It is also involved in a physiological loop linking the activity of the hypothalamo-pituitary-adrenal axis and adipose tissue. At the adrenal level, leptin has been shown to antagonize the effects of ACTH on glucocorticoid biosynthesis by decreasing the expression of various enzymes of the steroid biosynthetic pathway. The steroidogenic acute regulatory protein regulates cholesterol delivery to the P450(scc) enzyme, a process that is rate limiting in steroid hormone biosynthesis. We have demonstrated here that leptin significantly inhibits the expression of steroidogenic acute regulatory protein in primary cultures of rat adrenocortical cells. This inhibition was observed at both the protein and mRNA levels. In contrast, leptin was not found to interfere with the expression of the cytosolic enzyme cholesterol ester hydrolase or with that of the mitochondrial enzyme P450(scc). In addition, we observed the anticipated stimulation of cAMP production by ACTH in the presence of leptin, suggesting that it does not interfere with intracellular ACTH signaling. In summary, our data provide evidence that the interplay existing between leptin and ACTH in vivo is mediated at least partially via a direct and opposite modulation of steroidogenic acute regulatory protein, a key factor in the adrenal steroid biosynthetic pathway. This effect of leptin could also be relevant to other steroidogenic tissues.

Angiotensin II and calcium channels.

Sixty years after its initial discovery, the octapeptide hormone angiotensin II (AngII) has proved to play numerous physiological roles that reach far beyond its initial description as a hypertensive factor. In spite of the host of target tissues that have been identified, only two major receptor subtypes, AT1 and AT2, are currently fully identified. The specificity of the effects of AngII relies upon numerous and complex intracellular signaling pathways that often mobilize calcium ions from intracellular stores or from the extracellular medium. Various types of calcium channels (store- or voltage-operated channels) endowed with distinct functional properties play a crucial role in these processes. The activity of these channels can be modulated by AngII in a positive and/or negative fashion, depending on the cell type under observation. This chapter reviews the main characteristics of AngII receptor subtypes and of the various calcium channels as well as the involvement of the multiple signal transduction mechanisms triggered by the hormone in the cell-specific modulation of the activity of these channels.

La prevención del parto prematuro. Nuevas perspectivas / Prediction of preterm delivery: new approaches

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Blood flow velocity waveforms from fetal peripheral pulmonary arteries in pregnancies with preterm premature rupture of the membranes: relationship with pulmonary hypoplasia.

OBJECTIVES: To measure fetal peripheral pulmonary artery velocity waveforms by Doppler ultrasonography in pregnancies complicated by premature rupture of membranes under 24 weeks' gestation and to relate the Doppler indices to the development of fetal pulmonary hypoplasia. DESIGN: A prospective longitudinal study of fetal peripheral pulmonary artery velocity waveforms from premature rupture of membranes to delivery. SUBJECTS: Twenty pregnancies complicated by premature rupture of membranes before 24 weeks of gestation and delivering after 26 weeks. METHODS: Peripheral pulmonary artery velocity waveforms were recorded by Doppler technique at weekly intervals until delivery and Pulsatility Index (PI) calculated. Pregnancies were managed conservatively according to an institutional management protocol. Pulmonary hypoplasia was defined at autopsy by lung/body weight ratios and radial alveolar counts. Pulsatility Indices of fetuses developing pulmonary hypoplasia were compared with those with a normal lung development. RESULTS: After premature rupture of membranes PI values were higher than normal reference limits for gestation, but no differences were found between the six fetuses which developed pulmonary hypoplasia and the remaining 14 fetuses with normal lung development. In this latter group PI values progressively decreased with advancing gestation (ANOVA for repeated measurements F = 11.61; P < or = 0.001), while they increased in fetuses developing pulmonary hypoplasia (F = 8.44; P < or = 0.001). As a consequence of these opposite trends significant differences in PI values were present between the two groups of fetuses from 2 weeks after the premature rupture of membranes. Two weeks after the premature rupture of membranes a PI value from the peripheral pulmonary arteries above the 95th centile had a sensitivity of 62.5%, specificity of 94.6%, positive predictive value of 83.3%, negative predictive value of 78.5% and relative risk of 3.88 (95th confidence interval 1.34-11.28) for the prediction of pulmonary hypoplasia. CONCLUSION: The measurement of peripheral pulmonary velocity waveforms may help to establish the risk of developing pulmonary hypoplasia in pregnancies complicated by premature rupture of membranes.

The role of tyrosine kinases in capacitative calcium influx-mediated aldosterone production in bovine adrenal zona glomerulosa cells.

In adrenal glomerulosa cells, the stimulation of aldosterone biosynthesis by angiotensin II (Ang II) involves the activation of a capacitative Ca(2+) influx through calcium release-activated calcium (CRAC) channels. In various mammalian cell systems, it has been shown that CRAC channel activation and Ca(2+) entry require tyrosine kinase activity. We have therefore examined in this work whether similar mechanisms contribute to Ang II-induced mineralocorticoid biosynthesis. In fluo-3-loaded isolated bovine glomerulosa cells, two inhibitors of tyrosine kinases, genistein and methyl-2, 5-dihydroxycinnamate (MDHC) (100 microM) prevented capacitative Ca(2+) entry elicited by Ang II (by 54 and 62% respectively), while the inhibitor of epidermal growth factor (EGF) receptor tyrosine kinase, lavendustin A, was without effect. Similar results were observed on Ca(2+) influx triggered by thapsigargin, an inhibitor of microsomal Ca(2+) pumps. The inhibitors blocked Ang II-stimulated pregnenolone and aldosterone production in the same rank order. In addition to its specific effect on capacitative Ca(2+) influx, genistein also affected the late steps of the steroidogenic pathway, as shown by experiments in which the rate-limiting step (intramitochondrial cholesterol transfer) was bypassed with 25-OH-cholesterol (25-OH-Chol), cytosolic calcium was clamped at stimulated levels or precursors of the late enzymatic steps were supplied. In contrast, genistin, a structural analogue of genistein devoid of tyrosine kinase inhibitory activity, was almost without effect on pregnenolone or 11-deoxycorticosterone (DOC) conversion to aldosterone. These results suggest that, in bovine adrenal glomerulosa cells, Ang II promotes capacitative Ca(2+) influx and aldosterone biosynthesis through tyrosine kinase activation.

Measurement of perimitochondrial Ca2+ concentration in bovine adrenal glomerulosa cells with aequorin targeted to the outer mitochondrial membrane.

Microdomains of high cytosolic free Ca(2+) concentration in the proximity of mitochondria might have an important role in the stimulation of steroidogenesis in bovine adrenal glomerulosa cells. In the present study we have investigated local changes of free Ca(2+) concentration near the outer mitochondrial membrane ([Ca(2+)](om)) under stimulation with angiotensin II (Ang II) and K(+). Glomerulosa cells in primary culture were transfected with a recombinant cDNA encoding the N-terminal region of the human translocase protein 20 of the outer mitochondrial membrane, in frame with the Ca(2+)-sensitive photoprotein aequorin. This chimaeric aequorin (TomAeq) was associated with mitochondria-enriched subcellular fractions of transfected COS-7 cells and was susceptible to proteinase K, showing that it was targeted to the outer mitochondrial membrane, facing the cytosolic space. In bovine adrenal glomerulosa cells transfected with TomAeq cDNA, Ang II induced a transient [Ca(2+)](om) peak reaching 1.42+/-0.28 microM, which decreased immediately to the basal resting value. The peak response to Ang II was strikingly lower than the peak response of mitochondrial free Ca(2+) concentration, which increased to 5.4+/-1.2 microM. The smaller response of [Ca(2+)](om) to Ang II compared with the elevated matrix response did not result from buffering effects of the organelle, from altered mechanisms of intramitochondrial Ca(2+) transport or from differences in the affinity of the chimaeric aequorins for Ca(2+). This approach has allowed us to follow perimitochondrial Ca(2+) homeostasis in bovine glomerulosa cells under stimulation with Ca(2+)-mobilizing agonists and to reveal a strong gradient of Ca(2+) concentration between the mitochondrial matrix and the immediate environment of the organelle.

Angiotensin II negatively modulates L-type calcium channels through a pertussis toxin-sensitive G protein in adrenal glomerulosa cells.

In bovine adrenal glomerulosa cells, angiotensin II and extracellular K+ stimulate aldosterone secretion in a calcium-dependent manner. In these cells, physiological concentrations of extracellular potassium activate both T-type (low threshold) and L-type (high threshold) voltage-operated calcium channels. Paradoxically, the cytosolic calcium response to 9 mM K+ is inhibited by angiotensin II. Because K+-induced calcium changes observed in the cytosol are almost exclusively due to L-type channel activity, we therefore studied the mechanisms of L-type channel regulation by angiotensin II. Using the patch-clamp method in its perforated patch configuration, we observed a marked inhibition (by 63%) of L-type barium currents in response to angiotensin II. This effect of the hormone was completely prevented by losartan, a specific antagonist of the AT1 receptor subtype. Moreover, this inhibition was strongly reduced when the cells were previously treated for 1 night with pertussis toxin. An effect of pertussis toxin was also observed on the modulation by angiotensin II of the K+ (9 mM)-induced cytosolic calcium response in fura-2-loaded cells, as well as on the angiotensin II-induced aldosterone secretion, at both low (3 mM) and high (9 mM) K+ concentrations. Finally, the expression of both Go and Gi proteins in bovine glomerulosa cells was detected by immunoblotting. Altogether, these results strongly suggest that in bovine glomerulosa cells, a pertussis toxin-sensitive G protein is involved in the inhibition of L-type channel activity induced by angiotensin II.