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Plasma free and sulfoconjugated catecholamines were measured 28 times in 19 healthy men who had been in a recumbent position for at least 60 min (mean ± s.e.m., pg/ml): free dopamine = 36 ± 8, sulfoconjugated dopamine = 5568 ± 544, free norepinephrine = 178 ± 11, sulfoconjugated norepinephrine = 645± 29, free epinephrine = 37 ± 4, and sulfoconjugated epinephrine = 451 ± 26. They were also measured during passive and active upright posture, during extracellular volume expansion induced by an intravenous infusion of sodium chloride, and during recumbency, as a control, to investigate whether the free sulfoconjugated relationship might have a physiological role. Data obtained do not support such a hypothesis, even during episodes of orthostatic hypotension. In the control group, there was a progressive decrease of sulfoconjugated catecholamines during recumbency, between 8 a.m. and 12 noon. [ABSTRACT FROM PUBLISHER]

Spintronic terahertz emitters (STEs) are gapless, ultrabroadband terahertz sources that can be driven within a wide pump-wavelength and repetition-rate range. While STEs driven by strong pump lasers operating at kilohertz repetition rates excel in generating high electric field strengths for terahertz spectroscopy or ellipsometry, newly advancing technologies such as ultrafast modulation of terahertz polarization, scanning tunneling microscopy, laser terahertz emission nanoscopy, and fully fiber-coupled integrated systems demand an STE pumping at megahertz repetition rates. In all these applications the available terahertz power is ultimately limited by the STE's optical damage threshold. However, to date, only very few publications have targeted this crucial topic and investigations beyond the kilohertz repetition-rate regime are missing. Here, we present a complete study of our single-mode fiber-tip STEs' optical damage thresholds covering the kilohertz, megahertz, and gigahertz repetition-rate regimes as well as continuous-wave irradiation. As a very important finding, we introduce the necessity of classifying the optical damage threshold into two regimes: a low-repetition-rate regime characterized by a nearly constant fluence threshold, and a high-repetition-rate regime characterized by an antiproportional fluence dependence ("average-power threshold"). For our single-mode fiber-tip STEs, the transition between these regimes occurs around 4 MHz. Moreover, we present a cohesive theory of the damaging thermodynamical processes at play and identify temperature-driven inter-layer diffusion as the primary cause of the STE failure. These findings are substantiated by atomic force microscopy, infrared scattering-type scanning near-field optical microscopy, and scanning transmission electron microscopy measurements. This new level of understanding offers a clear optimization lever and provides valuable support for future advancements in the promising field of spintronic terahertz emission.

Spintronic terahertz emitters promise terahertz sources with an unmatched broad frequency bandwidth that are easy to fabricate and operate, and therefore easy to scale at low cost. However, current experiments and proofs of concept rely on free-space ultrafast pump lasers and rather complex benchtop setups. This contrasts with the requirements of widespread industrial applications, where robust, compact, and safe designs are needed. To meet these requirements, we present a novel fiber-tip spintronic terahertz emitter solution that allows spintronic terahertz systems to be fully fiber-coupled. Using single-mode fiber waveguiding, the newly developed solution naturally leads to a simple and straightforward terahertz near-field imaging system with a 90%-10% knife-edge-response spatial resolution of 30 µm.

Pasquier, Mathieu, Louis Marxer, Hervé Duplain, Vincent Frochaux, Florence Selz, Pierre Métrailler, Grégoire Zen Ruffinen, and Olivier Hugli. Indications and outcomes of helicopter rescue missions in alpine mountain huts: A retrospective study. High Alt Med Biol 18:355-362, 2017., Aims: This retrospective study describes the rescue indications and outcome of patients rescued by helicopter from mountain huts in the Swiss Alps. The hospital course and operational data were also studied., Results: Among 14,872 helicopter rescue missions undertaken during the 10-year study period, 309 (2.1%) were performed from mountain huts at a mean altitude of 2794 ± 459 m. The mean age of the patients was 43 ± 16 and 66% were male. Thirty-four percent of the patients had a National Advisory Committee for Aeronautics score ≥3. Most (89%) patients were transported to hospital and only 12 (3.9%) patients had to stay more than 48 hours. Hospital diagnoses were extremely varied. Trauma accounted for 50% and altitude diseases for 7% of the cases. A winching procedure was performed 18 times and 19 missions included a night flight., Conclusions: Helicopter rescue missions in mountain huts are a small part of all rescue missions. Our study provides a better understanding of medical emergencies arising in mountain huts. The diagnoses encountered are extremely varied in their type and severity. Hut keepers should be prepared for these situations as they will often have to act as first responders in the case of medical problems.

Background: Impaired Fas-induced apoptosis of lymphocytes in vitro is a principal feature of the autoimmune lymphoproliferative syndrome (ALPS). We studied six children with ALPS whose lymphocytes had normal sensitivity to Fas-induced apoptosis in vitro., Methods: Susceptibility to Fas-mediated apoptosis and the Fas gene were analyzed in purified subgroups of T cells and other mononuclear cells from six patients with ALPS type III., Results: Heterozygous dominant Fas mutations were detected in the polyclonal double-negative T cells from all six patients. In two patients, these mutations were found in a fraction of CD4+ and CD8+ T cells, monocytes, and CD34+ hematopoietic precursors, but not in hair or mucosal epithelial cells., Conclusions: Somatic heterozygous mutations of Fas can cause a sporadic form of ALPS by allowing lymphoid precursors to resist the normal process of cell death., (Copyright 2004 Massachusetts Medical Society)

Considerable progress has been recently accomplished in the management of patients who have undergone haplo-incompatible haematopoietic stem cell transplantation (HSCT) in terms of intake and prevention of graft-versus-host disease. Nevertheless haplo-incompatible HSCT is a procedure limited to a small number of patients because of the long-lasting immunodeficiency that is responsible for more than 50% of deaths within the first 3 months. Interleukin (IL)-7, which plays a unique and key role in T-cell development both in the mouse and in the human, is particularly attractive for attempting to speed up T-cell reconstitution. However, controversial results have been obtained after bone marrow graft in murine and primate models. To elucidate the impact of IL-7 treatment, we have performed HSCT in irradiated murine recombination activating gene (RAG) immunodeficient recipients, using donors that exhibited increased major histocompatibilty complex (MHC) incompatibility. Although irradiation performed prior to HSCT lead to a profound defect in the thymic stromal cells responsible for IL-7 production, IL-7 treatment had no significant effect on immune reconstitution in the MHC compatible and partially compatible settings. Interestingly, in the MHC fully incompatible setting in which only one-third of the recipients demonstrated active thymopoiesis, probably because of the rejection of donor cells by host natural killer cells, IL-7 treatment had a beneficial effect on T-cell development.

Affinity maturation of the Ab repertoire in germinal centers leads to the selection of high affinity Abs with selected heavy chain constant regions. Ab maturation involves two modifications of the Ig genes, i.e., somatic hypermutation and class switch recombination. The mechanisms of these two processes are not fully understood. As shown by the somatic hypermutation and class switch recombination-deficient phenotype of activation-induced cytidine deaminase (AID)-deficient patients (hyperIgM type 2 syndrome) and mice, both processes require the AID molecule. Somatic DNA modifications require DNA breaks, which, at least for class switch recombination, lead to dsDNA breaks. By using a ligation-mediated PCR, it was found that class switch recombination-induced dsDNA breaks in S mu switch regions were less frequent in AID-deficient B cells than in AID-proficient B cells, thus indicating that AID acts upstream of DNA break induction.

MHC class II deficiency is a combined immunodeficiency caused by defects in the four regulatory factors, CIITA, RFXANK, RFX5 and RFXAP, that control MHC II expression at the transcriptional level. The RFXANK gene encodes one subunit of the heterotrimeric RFX complex that is involved in the assembly of several transcription factors on MHC II promoters. Seven different RFXANK mutations have previously been reported in 26 unrelated patients. The most frequent mutation, a 26-bp deletion (752delG-25), has been identified in 21 patients. The other mutations are all nonsense or splice-site mutations, leading to proteins lacking all or part of the RFXANK ankyrin repeat region. We report two novel missense mutations, D121V and R212X, resulting in loss of function of the gene. We investigated the in vivo effects of these mutations and of three other point mutations on the expression of the RFXANK RNA and protein. The number of RFXANK transcripts was severely reduced in all patients except one. The RFXANK protein was barely detected in two cases. In addition, guided by a structural model of RFXANK, we investigated experimental mutants of the C-terminal tyrosine 224. Substitution Y224A, but not Y224F, led to the loss of function of RFXANK. Two null mutants, D121V and Y224A, were tested in protein interaction and DNA binding assays. The D121V mutant was unable to form the RFX complex, indicating that D121 is required for RFXAP binding. The Y224A mutant formed an RFX complex that bound normally to the MHC II promoter, but did not lead to MHC class II expression, whereas Y224F RFXANK retained the wild-type function. This indicates that an aromatic ring, but not the phenyl chain of tyrosine, is necessary at position 224 for normal RFXANK function. Studies on the Y224A mutant suggest that, in addition to the RFX subunits and CIITA, another protein is essential for MHC class II expression. This protein appears to interact with the fourth ankyrin repeat of RFXANK.

The expression of MHC class II molecules is essential for all Ag-dependent immune functions and is regulated at the transcriptional level. Four trans-acting proteins control the coordinate expression of MHC class II molecules: class II trans-activator (CIITA), regulatory factor binding to the X box (RFX)-associated protein; RFX protein containing ankyrin repeats, and RFX5. In humans, defects in these genes result in MHC class II expression deficiency and cause combined immunodeficiency. Most patients with this deficiency suffer from severe recurrent infections that frequently lead to death during early childhood. We investigated three sisters, now ages 21, 22, and 24 years, in whom MHC-II deficiency was detected. Even though the eldest sibling was asymptomatic and the other two had only mild immunodeficiency, none of the three class II isotypes was expressed on T cell blasts, fibroblasts, EBV B cell lines, or epidermal dendritic cells. Residual HLA-II expression was detected in fresh PBMC. Somatic complementation identified the disease as CIITA deficiency. A homozygous T1524C (L469P) substitution was found in the coding region of the CIITA cDNA and was shown to be responsible for the defect in MHC-II expression. This missense mutation prevents the normal functioning of MHC-II but does not lead to the nuclear exclusion of the L469P CIITA. Transfection experiments demonstrated that the CIITA L469P mutant had residual MHC class II trans activation activity, which might explain the unusual clinical course of the patients studied. This study shows that an attenuated clinical phenotype or an asymptomatic clinical course can be observed in patients despite a profound defect in the expression of MHC class II genes. The frequency of the inherited MHC class II deficiency might thus be underestimated.

Severe combined immunodeficiency-X1 (SCID-X1) is an X-linked inherited disorder characterized by an early block in T and natural killer (NK) lymphocyte differentiation. This block is caused by mutations of the gene encoding the gammac cytokine receptor subunit of interleukin-2, -4, -7, -9, and -15 receptors, which participates in the delivery of growth, survival, and differentiation signals to early lymphoid progenitors. After preclinical studies, a gene therapy trial for SCID-X1 was initiated, based on the use of complementary DNA containing a defective gammac Moloney retrovirus-derived vector and ex vivo infection of CD34+ cells. After a 10-month follow-up period, gammac transgene-expressing T and NK cells were detected in two patients. T, B, and NK cell counts and function, including antigen-specific responses, were comparable to those of age-matched controls. Thus, gene therapy was able to provide full correction of disease phenotype and, hence, clinical benefit.

Murine gut intraepithelial (IEL) T cell receptor (TCR)-alpha/beta lymphocytes bearing CD8alpha/13 or CD8alpha/alpha coreceptors have been shown previously to express different oligoclonal TCR beta chain repertoires in the same mouse, in agreement with other evidence indicating that these two populations belong to different ontogenic lineages, with only CD8alpha/beta+ IELs being fully thymus dependent. CD8alpha/beta+, but not CD8alpha/alpha+, T lymphocytes are also present in the lamina propria. Here, we show that CD8alpha/beta+ lymphocytes from the lamina propria and the epithelium are both oligoclonal, and that they share the same TCR-beta clonotypes in the same mouse, as is also the case for CD4alpha T cells. Furthermore, identical T cell clones were detected among CD8alpha/beta IELs and CD8alpha/beta+ blasts circulating into the thoracic duct (TD) lymph of the same mouse, whereas TD small lymphocytes are polyclonal. These findings must be considered in light of previous observations showing that T blasts, but not small T lymphocytes, circulating in the TD lymph have the capacity of homing into the gut epithelium and lamina propria. These combined observations have interesting implications for our understanding of the recirculation of gut thymus-dependent lymphocytes and their precursors, and of the events leading up to the selection of their restricted TCR repertoire.

We have generated two in vivo mouse models to study the regulation of DNA accessibility to the V(D)J recombinase machinery in the T cell receptor (TCR)-J alpha locus. In recombination activating gene (RAG)-deficient mice, both injection of a TCR-beta chain transgene (RTB mice) or anti-CD3-epsilon treatment in vivo (RT3 mice) lead to the same phenotype with homogeneous thymocyte populations blocked at the CD4+ CD8+ double positive (DP) stage. At this developmental stage, the TCR-alpha rearrangements are about to start, and the TCR-J alpha locus is frozen in an accessible but yet unrearranged configuration in these mice. We show high level of TCR-alpha germ-line transcription in thymocytes from RTB and RT3 mice. Transcripts are skewed towards the 5' end of the TCR-J alpha locus, and the T early alpha (TEA) sterile transcript is predominant and therefore provides a useful marker for the TCR-J alpha locus opening. Analysis of the DNA methylation status reveals a global surmethylation of the TCR-J alpha locus in the thymus in comparison with non-lymphoid cells in these mice. We propose that hypermethylation of the locus could precede a progressive demethylation, providing a specific protective and regulatory role in the rearrangement events.

To address the role of the TEA germline transcription, which initiates upstream of the TCR-J alpha S, in the regulation of TCR-J alpha locus accessibility, we created a mouse in which this region has been removed by homologous recombination. Normal development of T alpha beta cells and the expression of other TCR alpha germline transcripts in TEA-/- mice ruled out an exclusive role for TEA in the overall accessibility of the J alpha cluster. However, the rearrangement of the most 5' J alpha (J alpha 61 to J alpha 53) was severely impaired, indicating that TEA may control the DNA accessibility of a particular J alpha window. Moreover, the relative usage of every J alpha segment was affected. These results are consistent with TEA acting as a "rearrangement-focusing" element, targeting the primary waves of V alpha-J alpha recombination to the most 5' J alpha S in an ongoing TCR-J alpha rearrangement model.

Gut thymo-dependent (CD8 alpha + beta + or CD4+) or -independent (CD8 alpha + beta -) intraepithelial lymphocytes (IEL) mediate cytotoxicity following T cell receptor (TCR)-CD3 signaling, but only TCR gamma delta + and alpha beta + thymo-independent IEL show cytotoxicity of natural killer (NK) and antibody-dependent cell-mediated cytotoxicity types. Moreover, TCR alpha beta + and gamma delta + thymo-independent IEL express NK receptors, and may therefore be referred to as NK-TIEL. NK-TIEL cytotoxicity is mediated through perforin, Fas, or both pathways. In contrast to that of other NK cells, this cytotoxicity is not negatively regulated by signals delivered through the recognition of major histocompatibility complex class I molecules. Thus, gut IEL include T cell subsets with unique specificities and functions, ontogenically distinct from other T cell lineages, which may increase the antigenic repertoire diversity of the immune system participating in the defense of the epithelial barrier.

We have previously shown by means of fluorescence microscopy that antigen-independent adhesion of resting CD4 T cells to EBV-transformed B cells can be down-regulated by ligand interaction with CD4. In this study we used flow cytometry analysis of conjugate formation to confirm these findings. No conjugates between resting CD4 + T cells and B cells were initially detected in the latter method, because flow velocity in the flow chamber induces hydrodynamic elongation forces which disrupt low-affinity conjugates. After forcing cell conjugation by low-speed centrifugation of T and B cells, conjugates became detectable although in smaller numbers than in fluorescence microscopy. "Forced" cell conjugates had similar characteristics to their unforced counterparts, i.e., 37 degrees C temperature dependency, mediation by LFA-1/ICAM-1 and CD2/LFA-3 pathways, and transiency. The latter characteristic was at least partly mediated by CD4/HLA class II interaction, since adhesion of CD4 + T cells to HLA class II-B cells was more stable. In addition, adhesion was inhibited by anti-CD4 antibodies but not by an HLA DR-derived peptide known to inhibit unforced CD4 + T cell adhesion to B cells. This blocking effect was partially reproduced by reducing the centrifugation time prior to the adhesion assay. These results show that a) CD4-mediated down-regulation of T cell adhesion can be observed by means of two different techniques, and b) analysis of cell-cell adhesion after increasing centrifugation times (and possibly speeds) is a simple way of measuring adhesion forces semiquantitatively.

Dendritic cells (DC) are the main antigen-presenting cells for the initiation of primary T cell-mediated immune responses. In the first stage of activation, T cells bind to DC in an antigen-independent manner. We studied the adhesion characteristics of human CD4+ T cells to DC generated from CD34+ hematopoietic progenitors following 12 to 13 days of culture in the presence of granulocyte/macrophage colony-stimulating factor and tumor necrosis factor-alpha. A majority of these cells had the morphology, phenotype and functions of DC. CD4+ T/DC adhesion was measured by means of fluorescence microscopy and flow cytometry. Four independent receptor/ligand pathways, LFA-1/ICAM, ICAM/LFA-1, CD2/LFA-3 and CD28/CD80, were involved in the transient adhesion of DC to CD4+ T cells in antigen-independent and specific alloantigen-dependent situations, as shown by blocking experiments using monoclonal antibodies. The antibodies also blocked a primary mixed lymphocyte reaction (MLR) in which DC were used as stimulatory cells. Adhesion of alloreactive CD4+ T cells to antigen-presenting DC was stronger than that of resting CD4+ T cells, while peak adhesion occurred after 5 and 20 min, respectively. The LFA-1 ligands involved in adhesion of resting CD4 T cells to DC and alloreactive CD4+ T cells to specific DC differed in part, since ICAM-3 on resting T cells and ICAM-1 on alloreactive T lymphocytes preferentially bound LFA-1. Studies of interactions between DC and phorbol ester-activated T cells expressing the CD40 ligand revealed a fifth independent adhesion pathway, CD40/CD40 ligand. CD4-mediated regulation of CD4+ T/DC adhesion was suggested by the observation that preincubation of CD4+ T cells and DC individually with anti-CD4 antibodies inhibited adhesion. In addition, antibodies specific for HLA class II molecules inhibited adhesion when used to pretreat DC but not alloactivated CD4+ T cells.

The human T cell receptor was studied using an anchored-polymerase chain reaction (A-PCR) and hybridization with V beta-specific oligonucleotide probes, together with the few anti-V beta monoclonal antibodies (mAb) available. After confirming the semiquantitative and reproducible nature of the A-PCR technique, we assessed the complete V beta repertoire in sorted CD4+ and CD8+ lymphocyte populations from three normal donors. These experiments confirmed the absence of V beta-restricted deletions in human peripheral cells, in contrast to several mouse strains. This feature makes it difficult to study negative selection in man, given the apparent absence of an endogenous superantigen corresponding to the Mls system in the mouse. To investigate human V beta repertoire shaping, we studied V beta usage in CD4+ and CD8+ T cells from children with an inherited immunodeficiency characterized by defective expression of human leukocyte antigen class II molecules. An initial study using anti-V beta monoclonal antibodies failed to show significant abnormalities in V beta usage. Four patients analyzed using the A-PCR method all had a polyclonal V beta repertoire, suggesting normal positive selection and raising questions as to the importance of V beta major histocompatibility complex (MHC) interactions and the role of thymic MHC density in shaping the V beta repertoire.

The presence of transcripts of the recombination activating gene RAG-1 was studied by in situ hybridization on selected populations of murine thymocytes, peripheral lymphocytes and gut intraepithelial lymphocytes (IEL), obtained by cell sorting. RAG-1 mRNA was found in a majority of "double-positive" (DP) thymocytes, but was absent in "single-positive" thymocytes and peripheral T lymphocytes. The only other T lineages in which about 10%-20% of the cells contained RAG-1 mRNA, and in smaller amounts, were "double-negative" (DN), T cell receptor (TcR) gamma delta- cortical thymocytes and gut CD3- IEL. These observations suggest that (a) the high expression of RAG-1 transcripts in DP thymocytes is related to the process of expansion-selection of these cells, probably accompanied by repeated TcR rearrangements, and that (b) in contrast, CD3- IEL from the gut (which are thymus independent) as well as some DN thymocytes undergo limited TcR rearrangement giving rise locally to TcR+ T cells without prior extensive process of local expansion-selection. A small percentage of peripheral B cells also contained RAG-1 mRNA, raising the possibility that this protein may also be involved in immunoglobulin class switching.

We have previously demonstrated that the monoclonal antibody (mAb)-induced modulation of CD3 and CD4 antigens from the surface of human peripheral blood lymphocytes is not dependent from protein kinase C activity (Thuillier et al., Eur. J. Immunol. 1990. 20:1197). In the present report we study the effect of genistein and of herbimycin A, two potent inhibitors of protein tyrosine kinases (PTK), on the mAb-induced modulation of CD3 and CD4 surface antigens. Both genistein and herbimycin inhibited the mAb-induced internalization of CD3 and, in contrast, facilitated that of CD4 antigen. These results indicate that the mAb-induced modulation of CD3 is essentially dependent on the PTK pathway, whereas PTK appear to negatively regulate the mAb-induced modulation of CD4.

We studied the effect of staurosporine, a potent inhibitor of protein kinase C (PKC) activity, on the phorbol ester- or monoclonal antibody (mAb)-induced modulation of CD3 and CD4 surface antigens. Staurosporine (10(-5) M) completely inhibited phorbol ester-induced modulation but had no effect on that induced by mAb. These results indicate that the down-regulation of CD3 and CD4 observed after activation of the cells by the corresponding mAb is independent from PKC-mediated phosphorylations, and thus that the activation of PKC is sufficient but not necessary to induce the modulation of CD3 and CD4 antigens.

Intestinal intraepithelial lymphocytes (IEL) were studied, after isolation in humans, for their surface antigens with a large variety of monoclonal antibodies. They show peculiar characteristics when compared with peripheral blood lymphocytes and intestinal lamina propria lymphocytes. Although a majority of human intraepithelial lymphocytes (IEL) express an alpha/beta type of T cell receptor (TcR), 13% express a gamma/delta TcR, a percentage which was significantly higher than that found in blood and in lamina propria. In contrast to observations in mice, there was no evidence that normal human TcR gamma/delta+ intestinal IEL might use preferential variable segments of gamma genes. About 10% of human intestinal IEL expressed the alpha chain but not the beta chain of CD8, thus resembling a subset of CD8 alpha+beta- IEL, which was recently described in mice and found to be of thymoindependent origin. In addition, 10% of human IEL had a unique phenotype of immature T cells, as they bore only CD7, but no other T cell or natural killer cell markers. Finally, even the major population of IEL which expressed the usual markers of the T cell lineage (CD3, TcR alpha/beta, CD2, CD4 or CD8 alpha/beta) differed from peripheral blood T lymphocytes by their peculiar expression of surface antigens associated with activation. Indeed, 80% of IEL were CD45R0+, CD45A-, but co-expression of CD11a, CD29 and LFA-3 was inconstant. In addition, 90% of IEL expressed HML-1.

A defect of conjugation may play a role in the elevated plasma free norepinephrine observed in patients with cirrhosis. Plasma free, sulfoconjugated, and glucuronoconjugated catecholamine concentrations were assessed in 15 patients with cirrhosis and in 15 age-matched control subjects. Plasma free norepinephrine and epinephrine levels were significantly higher in patients with cirrhosis (481 +/- 75 and 96 +/- 16 pg/ml, respectively) than in those of the control group (307 +/- 33 and 42 +/- 10 pg/ml, p less than 0.05 and p less than 0.01, respectively). Plasma free dopamine levels were similar in both groups. Sulfoconjugated catecholamines were the predominant form in plasma from both cirrhotic patients and control subjects. The ratio of conjugated to total catecholamines was similar in the two groups. Therefore, it is unlikely that a defect in conjugation of catecholamines is contributing to the excessive plasma free norepinephrine and epinephrine concentrations found in patients with cirrhosis. Moreover, in patients with cirrhosis, no significant relation was found between plasma conjugated catecholamines and the severity of liver disease. This study shows that cirrhosis does not induce alteration in conjugation of catecholamines and that hepatocellular function is not essential for conjugation of circulating catecholamines.

Using a radioenzymatic technique, the highest concentrations of free catecholamines were found in the duodenum, and the lowest in the liver of untreated rats. When compared to the antrum, the concentration of free dopamine was higher, and that of norepinephrine lower in the fundus. As far as conjugated catecholamines are concerned, the tissue concentrations of both sulfo- and glucurono-conjugates were usually low, and often non detectable, with an exception: the concentration of glucurono-conjugated dopamine was very high in the duodenum, ileum, and liver of untreated rats.

The metabolism of catecholamines (CA) in non-selected patients with chronic renal failure and under hemodialysis (CRFh) was studied by measuring the concentration of plasma free, sulfo- and glucuroconjugated CA, dopamine (DA), norepinephrine (NE), and epinephrine (EPI). Our data demonstrate a statistically significant increase of free DA and free NE concentration in CRFh, while that of free EPI was not different from controls. However a careful scrutiny of 35 individual data suggests that sub-groups of patients with either high normal or low plasma-free NE concentration could exist; this likely heterogeneity could be a good explanation for conflicting conclusions provided by previous reports. Suspecting that conjugated CA might be altered in CRFh, plasma sulfo- and glucuro-conjugated DA, NE and EPI were also measured. We have found a predictable and highly significant increase of sulfo-conjugated CA; plasma concentration of glucuro-conjugated DA and NE in CRFh was not different from controls, while that of glucuro-conjugated EPI was significantly increased. The physiological meaning, if any, of these new observations on conjugated CA cannot be assessed at the moment. The effects of hemodialysis were also investigated. Measurements on the arterial and on the venous line showed highly significant differences for tyrosine, free and sulfo-conjugated CA, and a lack of difference for glucuro-conjugated CA. Thus tyrosine, free and sulfo-conjugated CA were eliminated by the artificial kidney, but not glucuro-conjugated amines.(ABSTRACT TRUNCATED AT 250 WORDS)

Free, sulfo and glucuro-conjugated catecholamines (dopamine, noradrenaline and adrenaline) were measured to study their metabolism in 35 non-selected patients with chronic renal failure, and under hemodialysis for various periods of time. Our data demonstrate a statistically significant increase of free dopamine, and free noradrenaline concentration in these patients, while that of free adrenaline was not different from controls. However a careful scrutiny of 35 individual data suggests that sub-groups of patients with either high normal or low plasma free noradrenaline concentration could exist; this likely heterogeneity could be a good explanation for conflicting conclusions provided by previous reports. Suspecting that conjugated catecholamines might be altered in these patients, plasma sulfo and glucuro-conjugated amines were measured. We have found a predictable and highly significant increase of sulfo-conjugated catecholamines; glucuroconjugated dopamine and noradrenaline were unchanged, while glucuroconjugated adrenaline was significantly increased. The physiological meaning, if any, of these new observations on conjugated catecholamines cannot be assessed at the moment.

The plasma catecholamine response to hypoglycemia was studied in a group of hypopituitary patients with Sheehan's syndrome before (group A) and after (group B) combined cortisol and thyroid hormone treatment as well as in a group of normal women (group C). The mean basal plasma norepinephrine (NE) level was significantly increased in group A compared to levels in groups B and C, in which values were similar. The mean basal plasma epinephrine (E) level was not significantly altered by hypopituitarism. The plasma NE response to hypoglycemia was similar in the three groups, while the plasma E response was blunted in groups A and B. However, the plasma E response was significantly decreased only in half of the patients. The basal E/NE ratio was similar in the three groups, but it was significantly decreased in groups A and B compared to that in group C at the peak. From these data we conclude that 1) hypopituitarism is characterized in the basal state by increased adrenergic tone, probably related to secondary hypothyroidism; and 2) during hypoglycemia adrenal stimulation is impaired only in some patients. The role of ACTH in the regulation of E secretion is minor. Impaired neurogenic regulation in some patients with Sheehan's syndrome could contribute to their illness.

Renal handling of free catecholamines (dopamine, norepinephrine and epinephrine) was studied in 36 hydropenic anesthetized mongrel dogs in accordance with the clearance technique. There was no statistically significant correlation between plasma concentrations and either systemic (mean blood pressure, or cardiac output) or renal (clearance of PAH or glomerular filtration rate) hemodynamics. Net tubular transport (NTT) was calculated as the difference between filtered load and urinary excretion for any catecholamine. The mean NTTs of free catecholamines were as follows: --2,72 ng/min for dopamine, --3,18 for norepinephrine, and --1,36 for epinephrine, showing that they are mostly reabsorbed. However the use of averages is misleading inasmuch as tubular transport of free catecholamines is a heterogeneous phenomenon: a secretion appears to predominate when plasma concentrations are low and a reabsorption predominates when they are high. Whether such a heterogeneity is due to either a genetic heterogeneity in mongrel dogs, or an age-related difference is discussed.