In situ characterization of antibody grafting on porous monolithic supports.

The efficient immobilization of antibodies on monolithic support is one of the most critical steps when preparing immunoaffinity supports. In this work, the ADECA (amino density estimation by colorimetric assay) method was adapted to tridimensional supports (in a dynamic mode) and proved to be efficient to characterize the antibodies grafting efficiency on 15.3±0.9mg porous glycidyl methacrylate (GMA)-co-ethylene dimethacrylate (EDMA) monolithic columns. The amount of grafted antibodies measured in situ on the monolith by ADECA (8.2±0.2µg of antibodies per milligram of monolith) was consistent with values obtained by bicinchoninic acid assay (BCA) after crushing the monolith. ADECA was shown to be less time-consuming and more versatile than BCA. The ADECA method was further implemented to thoroughly study and optimize the antibody grafting conditions (influence of pH and kinetics of the grafting step) on GMA-based monoliths and to check the covalent nature of the antibody/surface linking and its stability. Using the total amount of grafted antibodies and the amount of recognized antigen, we found that 65±6% of antibodies were able to capture their antigen. Finally, the grafting of Fab and F(ab')(2) fragments demonstrated that no significant improvement of the global binding capacity of the monolith was obtained.

Aminated dendritic surfaces characterization: a rapid and versatile colorimetric assay for estimating the amine density and coating stability.

The functionalization of surfaces with amino groups is used in many application areas such as in industrial biocatalytic processes for the development of medical biomaterials and in the environment for removing pollutants from water. Amino group density and grafting stability are often related to functionalized material performances; thus, their characterizations are of prime importance. The determination of amino density and grafting stability on polymeric material (e.g. polypropylene, polystyrene and cylco olefin copolymer) is often time consuming and sometimes presents technical constraints, more particularly with non-flat materials. In this paper, we report a novel colorimetric assay using the Coomassie Brilliant Blue dye for both amino density determination and grafting stability measurement. The assay named ADECA for "Amino Density Estimation by Colorimetric Assay" is sensitive, rapid, robust and versatile. We demonstrate that ADECA makes the evaluation of aminated materials performances possible for numerous material compositions, formats and chemistries used for grafting. Our study focuses on dendrigraft of poly-L-lysine and poly(amidoamine) dendrimers dendritic materials.

Functional characterization of the KNOLLE-interacting t-SNARE AtSNAP33 and its role in plant cytokinesis.

Cytokinesis requires membrane fusion during cleavage-furrow ingression in animals and cell plate formation in plants. In Arabidopsis, the Sec1 homologue KEULE (KEU) and the cytokinesis-specific syntaxin KNOLLE (KN) cooperate to promote vesicle fusion in the cell division plane. Here, we characterize AtSNAP33, an Arabidopsis homologue of the t-SNARE SNAP25, that was identified as a KN interactor in a yeast two-hybrid screen. AtSNAP33 is a ubiquitously expressed membrane-associated protein that accumulated at the plasma membrane and during cell division colocalized with KN at the forming cell plate. A T-DNA insertion in the AtSNAP33 gene caused loss of AtSNAP33 function, resulting in a lethal dwarf phenotype. atsnap33 plantlets gradually developed large necrotic lesions on cotyledons and rosette leaves, resembling pathogen-induced cellular responses, and eventually died before flowering. In addition, mutant seedlings displayed cytokinetic defects, and atsnap33 in combination with the cytokinesis mutant keu was embryo lethal. Analysis of the Arabidopsis genome revealed two further SNAP25-like proteins that also interacted with KN in the yeast two-hybrid assay. Our results suggest that AtSNAP33, the first SNAP25 homologue characterized in plants, is involved in diverse membrane fusion processes, including cell plate formation, and that AtSNAP33 function in cytokinesis may be replaced partially by other SNAP25 homologues.

[Disseminated histoplasmosis revealed by peripheral blood smear in an African immigrant with AIDS]. / Histoplasmose disséminée révélée par un frottis sanguin chez un immigré africain atteint du sida.

We report a case of disseminated histoplasmosis due to Histoplasma capsulatum var. capsulatum in an African patient with AIDS (born in Ghana) and living in France. This opportunistic fungal infection has never been reported in Ghana, maybe because of limited means of investigation or ignorance. Nevertheless, it was reported in a Ghanaian immigrant living in Italy. In our patient, the first diagnosis was suggested by a peripheral blood smear, which is not a classical biological diagnostic method. The diagnostic interest and the frequency of positive peripheral blood smears in the course of disseminated histoplasmosis are probably under estimated.

An arabidopsis T-DNA mutant affected in Nrt2 genes is impaired in nitrate uptake.

Expression analyses of Nrt2 plant genes have shown a strict correlation with root nitrate influx mediated by the high-affinity transport system (HATS). The precise assignment of NRT2 protein function has not yet been possible due to the absence of heterologous expression studies as well as loss of function mutants in higher plants. Using a reverse genetic approach, we isolated an Arabidopsis thaliana knock-out mutant where the T-DNA insertion led to the complete deletion of the AtNrt2.1 gene together with the deletion of the 3' region of the AtNrt2.2 gene. This mutant is impaired in the HATS, without being modified in the low-affinity system. Moreover, the de-regulated expression of a Nicotiana plumbaginifolia Nrt2 gene restored the mutant nitrate influx to that of the wild-type. These results demonstrate that plant NRT2 proteins do have a role in HATS.

Two-dimensional gel electrophoresis of proteins as a tool in wheat genetics.

In this minireview are reported several genetic investigations undertaken on wheat with the use of two-dimensional gel electrophoresis of total proteins extracted mainly from etiolated seedlings or from green leaves. Differences between developmental stages or organs of one genotype and nuclear and cytoplasmic genetic variations between genotypes are revealed by this method. We have also localized on the chromosomes structural genes coding for the proteins revealed and assigned their subcellular location to many polypeptides. We obtained new information concerning the regulation of protein amounts as well as the phylogenetic and homeology relationships between the A, B and D genomes.

Ambulatory blood pressure monitoring and postprandial hypotension in elderly persons with falls or syncopes.

BACKGROUND: Postprandial hypotension (PPH) is increasingly recognized as a common cause of falls and syncope in elderly persons. Noninvasive ambulatory blood pressure monitoring (ABPM) has been recommended for detecting PPH. This study investigates postprandial blood pressure (BP) changes by means of ABPM in elderly patients experiencing falls or syncopes. METHODS: Twenty-four-hour ABPM was performed in 156 inpatients (111 women, mean age 80.4 +/- 8.1 years). Among them, 45 had been admitted for falls and 75 for syncope; 36 with no history of falls or syncope served as controls. Postprandial change in systolic blood pressure (deltaSBP) was calculated by subtracting the mean SBP within the 2 hours following the meal from the mean SBP within the 2 hours preceding the meal. PPH was defined by a deltaSBP > or = 20 mm Hg. RESULTS: For the entire group, mean SBP decreased after the three meals. On average, the decline in SBP was greater after breakfast than after lunch or dinner, and the number of patients experiencing PPH was greater after breakfast. Average maximal deltaSBP was significantly larger in the syncope group than in the other groups ( p < .05). Moreover, the number of patients experiencing PPH was significantly higher in the syncope/fall group than in the control group (23% vs 9%; p = .03). Compared with patients without PPH, patients with PPH were more likely to have a history of diabetes mellitus (p < .01) or to use more than three different drugs daily ( p = .04), and they showed greater daytime SBP variability (p < .0001). Furthermore, there was a strong positive correlation between preprandial SBP and deltaSBP after breakfast. CONCLUSIONS: About one out of four elderly patients with falls or syncope experiences PPH, usually after breakfast. Postprandial decline in BP contributes to BP variability. deltaSBP and preprandial SBP are positively correlated.

Synthesis of full-length potyvirus cDNA copies suitable for the analysis of genome polymorphism.

New methods facilitating the synthesis and amplification of full-length cDNA copies of single-stranded viral RNA genomes have been developed. A method is described for the efficient purification of potyviral RNA and total RNA from infected plants and it is shown that they can serve as templates for the efficient synthesis of a full-length, 10 kb long, genomic cDNA. Two different reverse transcriptases were used (AMV-RT and MMLV-RT); only the first reverse transcriptase produced a good quality, full-length cDNA using viral RNA as a template. Surprisingly, MMLV-RT allowed for the full-length cDNA synthesis on virions rather than viral RNA. The PVY cDNA, synthesized using either RNA or virions, can be amplified successfully by PCR with high yields of full-length products. Such products are good substrates for the study by RFLP of the total genome polymorphism of virus isolates.

Des-enkephalin-gamma-endorphin in the treatment of schizophrenia.

Ninety-three patients with an exacerbation of chronic schizophrenia were included in a 4 week trial comparing placebo with 1, 3 and 10 mg des-enkephalin-gamma-endorphin (DE gamma E; beta-lipotrophin 66-77; Org 5878) per day (i.m.). Maintenance antipsychotic and other medications were continued unchanged. Treatment effects were assessed by means of the Comprehensive Psychopathological Rating Scale--subscale schizophrenia (CPRS-S), Brief Psychiatric Rating Scale (BPRS) and Global Assessment Scale (GAS) rating scales at weekly intervals. Safety data, i.e. laboratory investigations, vital signs and ECG recordings, were assessed before and during the trial. Side-effects were evaluated by means of a Record of Symptoms Emerging. Sixty-eight patients completed the trial, the reason for drop-out mainly being inadequate treatment effects and refusal of medication administration. One patient violated the protocol. After 4 weeks of treatment the mean CPRS-S score of the group receiving 10 mg DE gamma E daily had decreased statistically significantly more than the corresponding score of the placebo group (p less than 0.01). The same trend was apparent with BPRS (p = 0.08) and GAS (p greater than 0.1) scores. Therefore, the study should be considered inconclusive. No clinically relevant side-effects attributable to DE gamma E were observed.

[Systemic urticaria associated with autoimmune thyroiditis]. / Urticaire systémique associée à une thyroïdite auto-immune.

A case of chronic urticaria associated with thyroiditis is described. The diagnosis of autoimmune thyroid disease (Hashimoto's disease) rested on the presence of nodular goiter, thyroid dysfunction and significantly elevated thyroid microsomal antibodies (greater than 6,400). Skin biopsy showed changes suggestive of leucocytoclastic vasculitis. Immunological studies showed few abnormalities (low titers of antinuclear antibodies and rheumatoid factor) but a search for circulating immune complexes was negative, and serum complement levels were within normal range. The patient complained of severe pruritus and polyarthralgia but no systemic involvement occurred. Urticaria vasculitis has never previously been described in association with thyroid autoimmunity. This suggests the possibility of an autoimmune cause of urticaria. The urticaria improved and disappeared after treatment with levothyroxine. The frequent clinical latency of thyroiditis warrants systematic testing for circulating anti-microsome antibodies in women presenting with an apparently idiopathic chronic urticaria.

[Confusion in primary hyperparathyroidism. An uncommon manifestation in the elderly. 3 cases]. / Forme confusionnelle de l'hyperparathyroïdie primaire. Un mode de révélation particulier chez le sujet âgé. Trois observations.

Three cases of primary hyperparathyroidism disclosed by neuropsychiatric disorders in very elderly are reported. The frequency of these disorders and their almost constant association with muscle weakness are emphasized. Pre-operative imaging of the adenoma is often difficult. Surgical treatment is innocuous and considerably improves mental status.

[Methodological obstacles encountered in the evaluation of psychotherapies of schizophrenic patients. A general review]. / Les obstacles méthodologiques rencontrés dans l'évaluation des psychothérapies de schizophrènes. Revue générale.

Through a critical review of literature, authors suggest a methodological reflexion about problems in evaluation of effects of psychotherapies of schizophrenics, taking into account not only individual techniques but also institutional ones with psychotherapeutic orientations. Among the main difficulties are: evaluation with too much unspecific and over simplified criteria, for example criterium of rehospitalisation; lack of real comparability between groups, or techniques, and lack of validity of randomisation, still considered as a pawn of methodological rigour; diversity of levels of experience, and education of therapists, too unmatched model of functional organisation of the different centers, and in the same time too unequal recruitment of patients and at length of proposed therapeutic programs. Moreover, and even before these methodological bias, a more basic obstacle must be considered. It is the non-opening of models to the falsification, denounced by Bignami as the impossible verification of the heuristic value of the proposed models, ie the impossible verification of the utility of the proposed programs for individual therapies of schizophrenics.

[Use of benzodiazepines in the treatment of depression]. / Usage des benzodiazépines dans le traitement des dépressions.

The double-blind controlled trials between benzodiazepines and antidepressants show that benzodiazepines have not specific antidepressive action in depression. The benzodiazepines exert limited effect on the anxiety of endogenous and melancholia. They have definitive properties in neurotic depressions and mixed anxious-depressive states. In theses states, they may extenuate inhibition and elevate mood. Benzodiazepines are primarily anxiolytic rather than antidepressant -- But in some cases, anxiety and depression are not easily distinguished.

[Role of psychotherapy in the treatment of schizophrenia]. / Place des psychothérapies dans le traitement des schizophrénies.

Reality-adaptative, supportive psychotherapy, institutional cares, therapeutics with artistic mediations, family therapeutics are unanimously recognized as efficacious therapeutics for schizophrenic inpatients and outpatients. Conversely, the usefulness of psychoanalysis and exploratory, insight-oriented psychotherapy is questionnable. To clear up the question, we have studied controlled and non controlled studies comparing different psychotherapies, as well as the results of individual psychoanalysis. Among the controlled studies, Stanton's study stands out for its methological strictness. This study, in 164 schizophrenics, shows that insight-oriented psychotherapy is not more efficacious than supportive psychotherapy. In the continuation of this work, A. Frank and J. Gunderson show that the determinant factor of the psychotherapy's result is relational continuity and especially therapeutic alliance. Some works made in France show that 6 to 8 months are needed to appreciate the efficacy of institutional cares and psychotherapies can be observed. The psychoanalysis is not always efficacious. But these long acting experiences have provided irreplaceable knowledge towards the understanding of schizophrenic self and transference, effects of institutional cares, therapeutics with artistic mediations, supportive psychotherapy, theoretical reflection (Racamier, Rosenfeld, Searles, etc). Among the metapsychological knowledge, we must see: alterity's denial, struggle against thinking's activity, erotically paradoxical transference, receptacle's and depositary's function of therapists, inexistence of containing function. These knowledges explain that the two principal functions of the care given to schizophrenics are: help to self and presence.

[Study of the remission period (between crises) of unipolar and bipolar manic depressive disorders. Personal study of 95 patients]. / Etude de la période de rémission (intercrise) des troubles maniaco-dépressifs unipolaires et biopolaires. Etude personnelle de 95 patients.

The personality abnormalities observed during unipolar and bipolar manic-depressive disorders are known for years by german psychiatry as depressive temperament. In USA many works based on DSM III criteria have studied personality abnormalities coexisting with depression. The most frequently abnormalities are dependent, avoidant, compulsive, border line personality, observed with a frequency of 48% (P.A. Pilkonis) to 74% of patients (T.M. Shea et al.). But the personality traits or disorders are not the disorders only observed during remissions and intercrisis. Our personal work have studied the other disorders. This work concern pre-morbid period and intercrisis of 95 manic-depressed patients (45 unipolar and 50 bipolar patients). The time of remission is at least three months. Remission is certified by maximum scores of 10 on MADRS, 12 on Bech-Rafaelsen scale. The study of personality traits and other mental disorders is based on DSM III-R criteria. It is completed by Hamilton-Anxiety-Scale, Eysenck Personality Inventory and Beth-Israel Questionnaire of Sifneos. This study shows the following results. In pre-morbid period, the anxiety disorders are statistically more frequent in unipolar patients: generalized anxiety (35%), panic disorder (27%), psychosomatic disorders (30.5%). During intercrisis, the anxiety disorders are statistically more frequent in unipolar patients: generalized anxiety (51.1%), agoraphobia (31.1%) obsessive disorders (26.6%), panic disorder (18.2%). But this co-morbidity with anxious disorders is highly correlated with the incomplete characteristic of remission. During remission the personality abnormalities are similar to american studies, more frequent in unipolar patients, particularly obsessive personality (22.2 to 4%).(ABSTRACT TRUNCATED AT 250 WORDS)

[Effects of clonidine on opiate withdrawal symptoms. Results - biochemical mechanisms (author's transl)]. / Application de la clonidine au traitement de l'état de manque des toxicomanies opiacées. Résultats thérapeutiques, mécanismes d'action.

Clonidine was administered to nineteen patients in an inpatient setting after abrupt discontinuation of chronic opiate addiction (morphine, héroin, dextromoramide). Clonidine produces a decrease sometimes very rapid in opiate withdrawal signs but does not suppress the whole affects associated with. These data support the hypothesis that clonidine has antiwithdrawal effect by replacing opiate-mediated inhibition with alpha 2 mediated inhibition of brain noradrenergic activity.

[Plasma amino acid disturbances and depression]. / Perturbations des acides aminés plasmatiques et dépression.

Plasmatic amino acids concentrations were measured in 59 depressive patients and compared with that of a control population. Serine, glutamine, cystine, lysine and arginine have increased levels in the depressed males and females; so are 3 competitors of the tryptophan for the transport through the blood-brain barrier: valine, leucine and isoleucine. The tryptophan ratio is decreased in the depressive males. The sum of the competitors of the tryptophan is increased in the depressive males and females. All these results are discussed in relation with an abnormality of the carrier through the blood-brain barrier in the depressive state, without no proof thereof being however given.

[Initial results of a pharmacokinetic study of pipothiazine and its palmitic ester (Piportil L4) in a schizophrenic population]. / Premiers résultats d'une étude pharmacocinétique de la pipotiazine et de son ester palmitique (Piportil L4) dans une population de schizophrènes.

Plasma kinetics of pipotiazine have been studied in ten schizophrenic patients after oral administration of single dose of pipotiazine at 7 a.m. (30 mg, drops). Peak plasma concentrations are reached one hour after administration (41.8 +/- 19,9 ng/ml) and then rapidly decline until 24 h (2.2 +/- 1.2 ng/ml). During the following days plasma concentrations remain stable at the same sampling times. After 3 days wash-out and first intramuscular injection of pipotiazine palmitate (100 mg) main pharmacokinetic data are found: plasma concentrations of pipotiazine are not detectable during at less 3 days after injection, maximal drug level is attained during second week after i.m. (1.7 +/- 0.9 ng/ml). A period of decline is then recorded. Furthermore the mean ratio between pipotiazine plasma concentration after oral and i.m. administration is about 20. When pipotiazine palmitate is given every fourth week, steady state seems to be reached as early as the second month.

[Endogenous affective disorder, seasons of birth and photoperiodicity]. / Troubles de l'humeur endogènes, saisons de naissance et photopériodisme.

In a retrospective study, we compared the months of birth of 3,106 psychiatric inpatients to those of 1,943 surgical patients collected during the same period 1981-1991 in the same hospital, and of a sample of 10,003,572 births in France in 1977-1989. DSM III-R categories were modified so to allow a comparison with former studies, and psychiatric patients were distributed among seven categories: Bipolars (N = 294), Unipolars (N = 287), Neurotic-reactive depressions (N = 582), First Major Depressive episode (N = 214), Schizophrenia (N = 244), Schizo-Affectives (N = 52) and Other Diagnosies (N = 1,433). Months of birth were grouped in quarters and semesters, according to the usual calendar, but also to temperature and the photoperiodic cycle. The main results were: 1. A seasonnality of births in the General Population sample, with a spring maximum (p < 0.001). 2. An absence of deviation from the general population and the surgical sample among Neurotic-reactive Depressions and Other Diagnoses. 3. A deviation from the general population and from other comparison groups (surgical cases and Other Diagnoses) among Unipolars and First Major Depressive Episodes (most of those being late episodes), with a significant excess of births during the "dark" or "cold" season of the year, especially around the winter solstice. The Bipolar group followed the same tendency, though to a lesser degree and for subjects born before 1940 only. The most significant results were found among Unipolars, which differ from the general population either by quarters (p < 0.0005) or by semesters (p < 0.0005) and from surgical cases by quarters (p < 0.01) and by semesters (p < 0.001). The results were similar for First Major Depressive episodes, although this category was theoretically "anosological". As the median age was high in this category, it might group a number of late depressive episodes, near to "involutionnal melancholia". Thus, our results seem to be relevant to the traditional endogenous-psychogenetic dichotomy, with a "cold" or "dark" seasonnality of births in the first case, and no particular seasonnality in the second case. Some former studies showed the same results, but the most significant deviation was found in mania. Our results cannot be explained by differences in the sex-ratio among the categories, and only partially by an age-incidence effect or an age-cohort effect.(ABSTRACT TRUNCATED AT 400 WORDS)

[Study of antidiuretic hormones (arginine vasopressin) in 24 cases of primary adrenal insufficiency]. / Etude de l'hormone antidiurétique (arginine-vasopressine) dans 24 cas d'insuffisance surrénale primitive.

24 Addisonian patients were compared to 27 healthy subjects. Radioimmunoassay of plasma antidiuretic hormone showed higher concentrations of Arginine-vasopressin (AVP) in Addisonians than in normals, in spite of lower natremia and plasma osmolality. Statistical analysis showed a negative correlation between AVP and osmolality in Addisonian patients. On the other hand, no correlation was found between AVP and cortisol, or AVP and PRA. These results suggested an inadequate secretion of antidiuretic hormone in Addison disease. Under substitutive treatment, glucocorticoids alone didn't completely lower AVP concentration which was only normalized after administration of mineralocorticoids.