[Antipsychotic-drug-induced hyperprolactinemia: physiopathology, clinical features and guidance]. / Hyperprolactinémies induites par les antipsychotiques : physiopathologie, clinique et surveillance.

BACKGROUND: Hyperprolactinemia is a frequent but neglected adverse effect observed in patients treated with antipsychotic-drugs. In this review, we summarize its physiopathogenetic mechanism, its clinical manifestations in men and women, and the way to manage it. LITERATURE FINDINGS: Prolactin is a hormone secreted by lactotroph cells in the anterior pituitary. Its synthesis and release are under the control of peptides, steroids and neurotransmitters. The main inhibitory regulation is made by dopamine, which binds dopamine receptors D2 on the membrane of lactotroph cells. Antipsychotic-drugs block these receptors and thus remove the inhibitory effect of dopamine on prolactin secretion. All antipsychotic-drugs block D2 receptors and all can induce hyperprolactinemia. Nonetheless, it seems that the faster the antipsychotic-drug dissociates from D2 receptors, the lesser the increase of prolactin in the plasma. Another way to explain hyperprolactinemia is the ability of antipsychotic-drugs to cross the blood-brain barrier. The role of their metabolites should also be considered. For these reasons, one can distinguish prolactin-raising (conventional neuroleptics, amisulpride, risperidone) and prolactin-sparing (clozapine, aripiprazole, olanzapine) antipsychotics. An English study showed that 18% of men and 47% of women treated with antipsychotics for severe mental illness had a prolactin level above the normal range. Hyperprolactinemia is in fact more frequent in women than in men. Sometimes it is asymptomatic, but the higher the prolactin level is, the more patients have clinical manifestations. Some symptoms are due to the hypogonadism caused by prolactin, which disturbs hypothalamic-pituitary axis function, and others are due to direct effects on target tissues. Consequently, patients can suffer from sexual dysfunction, infertility, amenorrhea, gynecomastia or galactorrhoea. Data suggest that these symptoms are common, but patients don't mention them spontaneously and clinicians underestimate their prevalence. In the long-term, hypogonadism involves a premature bone loss in men and women. Klibanski and colleagues showed that this loss is significant only in women with hyperprolactinemia associated with amenorrhea. That suggests that prolactin is not directly responsible for this clinical feature. Nevertheless, prolactin seems to be involved in the development of breast cancer, but its role is unclear for prostate cancer. DISCUSSION: Our review promotes a check-up before beginning a treatment with antipsychotic agents. First, a baseline prolactin level should be measured. It should also include the research on previous treatment with antipsychotic-drugs and the assessment of adverse effects suggestive of hyperprolactinemia. Questioning should finally look for any contra-indication to antipsychotics. Monitoring during antipsychotic treatment has been studied by a group of international experts in psychiatry, medicine, toxicology and pharmacy who made a critical review of clinical guidance on hyperprolactinemia. Experts notify that it is important to check whether patients have any sexual dysfunction, such as loss of libido or menstrual irregularity, and galactorrhoea. Prolactin level should also be controlled after three months of stable dose treatment, or if any clinical feature of hyperprolactinemia appears. If a patient prescribed antipsychotic-drugs has a confirmed prolactin level above the normal range, it is necessary to exclude other causes of hyperprolactinemia. If antipsychotics are really involved, the management should be adapted with the prolactin level and the patient him/herself. To summarize, clinicians can decrease the dose of the antipsychotic or switch to a prolactin-sparing drug. Oral contraceptives can be added whether to prevent pregnancy or to prevent bone loss and osteoporosis. Finally, experts recommend reserving dopamine agonists to treat antipsychotic-induced hyperprolactinemia in very exceptional circumstances as it can worsen the mental illness.

Immunomodulatory effects of therapeutic preparations of human albumin.

BACKGROUND AND OBJECTIVES: Albumin is the most abundant protein in plasma and is considered to be immunologically inert. However, we recently observed that therapeutic human albumin preparations, used as protein control in studies involving high doses of IVIg, modulated the MHC II-restricted activation of antigen-specific T cells. In the present work, we characterized this effect in more details. MATERIALS AND METHODS: An in vitro antigen presentation assay using mouse cells was used to evaluate the effect of therapeutic human albumin preparations on the activation of ovalbumin-specific T cells. Flow cytometry and quantitative real-time PCR were used to monitor the expression of genes involved in this process. RESULTS: Therapeutic human albumin preparations increased T cell activation in a dose-dependent manner. The effect was explained by an increase in the expression of MHC II and of two other genes (CIITA and H2-M) involved in antigen presentation by murine monocytic cells. Similarly, the expression of HLA-DR on the surface of human monocytic cells was increased following incubation with therapeutic human albumin preparations. CONCLUSION: Altogether, these results reveal a possible physiological role of albumin in immunological processes, leading to an increased ability of antigen presenting cells to trigger T cell activation. This immunomodulatory effect needs to be considered, at least in studies in which albumin is used as a presumably inert control protein.

Genomewide identification of genes under directional selection: gene transcription Q(ST) scan in diverging Atlantic salmon subpopulations.

Evolutionary genomics has benefited from methods that allow identifying evolutionarily important genomic regions on a genomewide scale, including genome scans and QTL mapping. Recently, genomewide scanning by means of microarrays has permitted assessing gene transcription differences among species or populations. However, the identification of differentially transcribed genes does not in itself suffice to measure the role of selection in driving evolutionary changes in gene transcription. Here, we propose and apply a "transcriptome scan" approach to investigating the role of selection in shaping differential profiles of gene transcription among populations. We compared the genomewide transcription levels between two Atlantic salmon subpopulations that have been diverging for only six generations. Following assessment of normality and unimodality on a gene-per-gene basis, the additive genetic basis of gene transcription was estimated using the animal model. Gene transcription h(2) estimates were significant for 1044 (16%) of all detected cDNA clones. In an approach analogous to that of genome scans, we used the distribution of the Q(ST) values estimated from intra- and intersubpopulation additive genetic components of the transcription profiles to identify 16 outlier genes (average Q(ST) estimate = 0.11) whose transcription levels are likely to have evolved under the influence of directional selection within six generations only. Overall, this study contributes both empirically and methodologically to the quantitative genetic exploration of gene transcription data.

[Cardiological monitoring of antipsychotic-treated patients: evaluation and evolution of a hospital protocol]. / Surveillance cardiologique des patients traités par neuroleptiques : évaluation et évolution d'un protocole hospitalier.

INTRODUCTION: Following the Agence Française de Sécurité Sanitaire des Produits de Santé (Afssaps) (French Health Authority) recommendations in 2001, which impose the rigorous follow-up (electrocardiogram [ECG] and ionogram) of patients treated with antipsychotics (AP), a monitoring protocol was elaborated and set up in the Caen psychiatric hospital in April 2002. Protocol evaluation compared with fixed aims was performed after two years' follow-up. AIM OF THE STUDY: This protocol had to answer a triple aim: better identification of patients at risk, ensure in-treatment monitoring, be simple and adapted to daily practice. INCLUSION CRITERIA: A systematic admission check-up (S0) which includes cardiological and biological controls and after one and six months' treatment control (S1 and S6) were recommended. The major risk factors (RF) researched were long QT interval, bradycardia and hypokaliemia. RESULTS: The initial monitoring was conducted in 601 patients (that only corresponded to 17% of hospital admission active files during the considered period). Means delays before obtaining an ECG were three times those obtained existing biological check-ups (11 days versus three days after date of admission). Systematic and integrated characterisation controls on admission were not respected. We noted that two-third of patients admitted during this period were hospitalized for only five days, although the mean time to obtain an ECG is of 11 days. This delay (approximately one week) between ECG and biological check-up is not compatible with a complete patient RF evaluation. Respectively, 83 and 68 patients were controlled under treatment at S1 and S6. Only half of the patients were controlled at the one-month (S1) and 16% at the six-months' theoretical dates (S6). These delays are inappropriate, notably with regard to the mean time of hospitalisation (15-17 days). The incidence of major RF was higher in treated (71%) than in non-treated patients. Major RF presence at S0 was not systematically associated with an AP treatment contraindication. The excessive delay before the first ECG could partially explain why this initial check-up was not able to detect a pretreatment contraindication. On the other hand, AP treated patients who presented at least one major RF at S0 were more frequently monitored at S1 than patients who did not (26% versus 13%, p=0.05). Among the 168 patients treated with AP or other drugs prolonging the QT interval at risk, 33 had at least one follow-up. This risk population was not better controlled than the initial cohort. DISCUSSION: Protocol evaluation is essential to improve its interest and feasibility. If systematic characterisation is simple, its application in practice is very difficult. The second version of the protocol presented here proposes to substitute the systematic ECG characterisation with the classification on admission of patients in "risk groups" that will condition the subsequent monitoring. Risk groups are identified into two RF types: those which are not related to AP treatment and those which are therapeutic attitude is adapted according to initial QTcorrigé (QTc), its progression between S0 and S (seven days) and kaliemia.

Crystal-induced neutrophil activation. IV. Specific inhibition of tyrosine phosphorylation by colchicine.

We recently demonstrated that pathologically relevant inflammatory microcrystals, namely triclinic monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD) crystals, potently stimulate a characteristic protein tyrosine phosphorylation pattern in human neutrophils that differed from that observed in response to other soluble or particulate agonists. In this study, the effects of colchicine on protein tyrosine phosphorylation induced by MSU and CPPD crystals in human blood neutrophils were investigated. Immunoblot analysis with antiphosphotyrosine antibodies demonstrated that colchicine dose-dependently inhibited the tyrosine phosphorylation of all the proteins phosphorylated in response to MSU and CPPD crystals. Other microtubule-disruptive agents such as vinblastine, nocodazole, and colcemid also inhibited crystal-induced protein tyrosine phosphorylation while lumicolchicine and trimethylcolchicinic acid were without effect. Indomethacin and phenylbutazone were similarly without effect on microcrystal-induced tyrosine phosphorylation. Colchicine, as well as the other active alkaloids, failed to inhibit the protein tyrosine phosphorylation elicited by FMLP, C5a, leukotriene B4, and unopsonized zymosan. Overall, these results demonstrate that colchicine specifically and significantly inhibits the protein tyrosine phosphorylation induced by MSU and CPPD crystals and suggest that its effects are associated, at least in part, with its interaction with microtubules. Furthermore, the use of microtubule-disrupting drugs demonstrate that the mechanisms implicated in the induction of protein tyrosine phosphorylation by microcrystals differed from those involved in response to other soluble or particulate agonists.

Crystal-induced neutrophil activation. III. Inflammatory microcrystals induce a distinct pattern of tyrosine phosphorylation in human neutrophils.

The activation of human neutrophils by monosodium urate and calcium pyrophosphate dihydrate crystals is believed to play a critical role in the pathogenesis of arthritides such as acute gout and pseudogout, respectively. In this study, we investigated the potential involvement of tyrosine phosphorylation in microcrystal-mediated activation of human neutrophils. Immunoblot analysis with antiphosphotyrosine antibodies demonstrated that triclinic monosodium urate and calcium pyrophosphate dihydrate crystals stimulated a time- and concentration-dependent tyrosine phosphorylation of at least five proteins (pp130, 118, 80, 70, and 60). While phosphoprotein (pp) 118 and pp70 were the major phosphorylated substrates, pp70 was the dominant one in reactivity with antiphosphotyrosine antibodies. When the temporal patterns, as well as the levels of tyrosine phosphorylation for both types of crystals were compared, monosodium urate crystals were found to be more potent activators than calcium pyrophosphate dihydrate crystals. The tyrosine phosphorylation patterns induced by microcrystals differed from those stimulated by other soluble (FMLP, C5a, or leukotriene B4) or particulate (unopsonized latex beads or zymosan) agonists which stimulated preferentially the tyrosine phosphorylation of pp118. The ratio of the intensities of pp118 and pp70 were specific of the stimulation with microcrystals when compared to those observed with the other soluble or particulate agonists. Colchicine, a drug used specifically in the treatment of gout and pseudogout, inhibited microcrystal-induced tyrosine phosphorylation, while beta- and gamma-lumicolchicine were without effect. On the other hand, colchicine failed to inhibit FMLP-induced tyrosine phosphorylation. Furthermore, while colchicine inhibited the activation of the NADPH oxidase by microcrystals, it, on the other hand, enhanced the production of superoxide anions by FMLP. Taken together, these results (a) demonstrate that tyrosine phosphorylation is involved in the mechanism of activation of human neutrophils induced by microcrystals; and (b) suggest, on the basis of the characteristics of the observed patterns of tyrosine phosphorylation, that this response may be specific to the microcrystals and relevant to their phlogistic properties.

Activation of human neutrophils in vitro and dieldrin-induced neutrophilic inflammation in vivo.

Many chemicals of environmental concern are known to alter the immune system and are considered toxic molecules because they affect immune cell functions. Inflammation related to environmental chemical exposure, however, is poorly documented, except that from air pollutants. In this study, we found that the organochlorine insecticide dieldrin could not alter the ability of human neutrophils to phagocytose opsonized sheep red blood cells at nonnecrotic concentrations (0.1, 1, 10, and 50 microM). However, dieldrin was found to increase human neutrophil superoxide production, RNA synthesis, and proinflammatory cytokine interleukin-8 production. The normal apoptotic rate of neutrophils evaluated by both cytology and flow cytometry (CD-16 staining) was not altered by dieldrin treatments, and this was correlated with its inability to inhibit spreading of neutrophils onto glass. Using the murine air pouch model, we found that dieldrin induces a neutrophilic inflammation. Taken together, these results demonstrated that dieldrin is a proinflammatory contaminant. To our knowledge, this is the first report establishing that dieldrin is a contaminant exhibiting proinflammatory properties. In addition, it is the first time that the murine air pouch model has been successfully used to confirm that a chemical of environmental concern can induce an inflammatory response in vivo.

Paradoxical effects of colchicine on the activation of human neutrophilis by chemotactic factors and inflammatory microcrystal.

Neutrophil activation by chemotactic factors and by inflammatory microcrystals is accompanied by increases in protein tyrosine phosphorylation and by the activation of the NADPH oxidase. The addition of colchicine inhibited both responses induced by triclinic monosodium urate or calcium pyrophosphate crystals. On the other hand, colchicine enhanced the tyrosine phosphorylation of specific protein in neutrophils stimulated by chemotactic factor and augmented the production of superoxide anions induced by these same agonists. The effects of colchicine were shared by other anti-microtubule agents (nocodazole and vinblastine) but not by its inactive analogue beta-lumicolchicine, trimethylcolchicinic acid, indomethacin, or phenylbutazone. Furthermore, the (enhancing as well as inhibitory) effects of colchicine on tyrosine phosphorylation and superoxide anion production were reversed by taxol. Finally, in human cytoplasts colchicine again inhibited microcrystal-stimulated tyrosine phosphorylation but did not change chemotactic factor-stimulated phosphorylation. These data strongly support the hypothesis that microtubule-related mechanisms are involved in the modulation of the tyrosine phosphorylation response in human neutrophils, and suggest that a relationship may exist between the augmentation of tyrosine phosphorylation and of the stimulation of the NADPH oxidase induced by chemotactic factors.

In vitro leukotriene (LT) C4 synthesis by blood eosinophils from atopic asthmatics: predominance of eosinophil subpopulations with high potency for LTC4 generation.

We compared the leukotriene (LT) C4 generation by the eosinophil density subpopulations isolated from the blood of asthmatics and normal subjects. Using discontinuous Percoll gradients, eosinophil subpopulations with densities of 1.075, 1.078, 1.081, 1.084, 1.087 and 1.100 g/ml were isolated from the blood of six atopic asthmatics and seven normals. In normals, most eosinophils (94%) were recovered in the density fractions (1.084, 1.087, and 1.100 g/ml). In asthmatics, the eosinophil density profile was shifted towards lower cell density: the eosinophil subsets 1.078, 1.081 and 1.084 g/ml were increased 4.5, 30.3 and 8.9-fold respectively compared to normals (p less than 0.0001); the intermediate subsets, the hypodense 1.081 and normodense 1.084 g/ml fractions being the predominant subpopulations. The ability of asthmatic eosinophil subsets 1.078 to 1.087 g/ml to release LTC4 was measured by reversed-phase HPLC, LTC4 production was highest with the normodense 1.084 g/ml eosinophil subset (149 +/- 28 pmol/10(6) eosinophils, p less than 0.01 compared to the 1.081 fraction). The eosinophils from the hypodense 1.081 g/ml fraction also released more LTC4 (112 +/- 19 pmol/10(6) eosinophils) than the 1.078 and 1.087 g/ml fractions (70 +/- 16 and 67 +/- 12 respectively, p less than 0.01). These results show that, compared to normals, blood of mild atopic asthmatics contains an elevated number of eosinophils of intermediate density which have a high capacity for LTC4 production.

Stimulation of tyrosine hydroxylase activity by cyclic AMP in synaptosomes and in soluble striatal enzyme preparations.

Dibutyryl cyclic AMP (dB-cAMP) elicits a concentration-dependent stimulation of tyrosine hydroxylase activity in the striatal and mesolimbic synaptosomes. The per cent of stimulation is significantly higher in the mesolimbic synaptosomes than in the striatal synaptosomes. dB-cAMP and depolarizing agents (ouabain or veratridine) have an additive effect on synaptosomal tyrosine hydroxylase activity, indicating that they stimulate tyrosine hydroxylase activity by different mechanisms. cAMP does not stimulate soluble striatal tyrosine hydroxylase activity unless it is added in combination with ATP and Mg2+, compounds required for the activity of cAMP-dependent protein kinase. The cAMP elicited per cent stimulation of soluble tyrosine hydroxylase activity is dependent upon the concentration of added protein kinase and upon the pH of the reaction. dB-cAMP has the same effect on the kinetic state of tyrosine hydroxylase in synaptosomes as cAMP on the soluble tyrosine hydroxylase. The nucleotide does not alter the apparent Km for tyrosine, reduces the Km for the pteridine cofactor and increases the Ki for dopamine. Thus, cAMP increases the affinity of tyrosine hydroxylase for the pteridine cofactor and concomitantly decreases the affinity for the end-product inhibition.

Familial periodic ataxia responsive to acetazolamide.

Two cases, a father and son, of recurrent cerebellar ataxia in the same family are reported, suggesting a familial trait for the dysfunction. In the older male the onset of each episode (30-90 min.) was signalled by dysarthria which then progressed towards gait ataxia; the son presented closely similar clinical symptoms. Physical examination and blood chemistry revealed no obvious neurological deficit or biochemical abnormalities, with the exception of I-III and III-IV evoked auditory wave interpeak latencies, which were found markedly abnormal on the left side in the father but not in the son; the EEG of both individuals showed some diffuse, slow wave abnormalities. A low dose of acetazolamide, 250 mg daily, has successfully repressed recurrence of the attacks over the past six months. Temporary withdrawal for 14 days of the carbonic anhydrase inhibitor in the father coincided with two observed ataxic episodes.

Death, dying, and the brain.

These cases have been cited and these issues presented to make the point that principles are often out of phase with perceptions. The argument is not that principles should cede to perceptions. This would imply resignation to the unlikelihood of moral development. Neither do we claim that perceptions should simply and generally capitulate to principles. This could mean an intolerable dictatorship of one philosophy over experience. Clinical ethics is the challenge to integrate principles and perceptions continuously and critically in resolving value conflicts about the best care of individual patients.

Rapid DNA purification for Hal gene PCR diagnosis in porcine tissues and extension to other meat species.

Two different halothane (Hal) gene polymerase chain reaction (PCR) tests were applied to genomic DNA extracted from porcine blood, semen, muscle and fat tissues by a rapid and simple Chelex-100 based method. One of the PCR procedure is designed from the ryanodine receptor coding sequence to produce a 81 base pair (bp) fragment, while the other is designed from pig intron sequences to produce a 659 bp fragment. Oligonucleotide primers derived from the coding sequence were also used for other meat species. Amplification products obtained from porcine, bovine, ovine, equine and deer genomic DNA were successfully digested with Hha I restriction enzyme to produce the same electrophoretic pattern as in the normal homozygous (NN) pig. No PCR products could be amplified from chicken and turkey DNA.

The effects of local anesthetics on postoperative pain.

This study was performed to determine if intraoperative local anesthesia improved control of postoperative pain after inguinal herniorrhaphy and to compare the effects of two commonly used local anesthetics on pain management. The Gate Control Theory of Pain formed the theoretical basis for this study. A retrospective nonexperimental study in an ex post facto design was used. Data were collected from 1990 through 1997 on 120 patient charts. The use of local anesthetic intraoperatively significantly decreased patients' lengths of stay postoperatively (P = 0.00) and need for postoperative narcotics (P = 0.00). Bupivacaine was found to be superior to lidocaine in decreasing the need for postoperative narcotic analgesia. Researchers concluded that many patients would benefit from intraoperative injection of local anesthesia. This information can affect patient care outcomes through decreasing recovery time, reducing postoperative pain, and reducing health care costs.

Site-directed mutagenesis of the polyomavirus genome: replication-defective large T mutants with increased immortalization potential.

We used the deletion loop mutagenesis procedure to direct point mutations into a small region of the polyomavirus genome, at 0.97 map units, affecting the structure of both the middle and large T antigens. We describe the construction of six middle T mutants which have retained the ability to transform rat cells in culture and four large T mutants, three of which are deficient in viral DNA replication and capable of immortalizing primary rat embryo fibroblasts very efficiently.

The differential effects of biofeedback in the treatment of menstrual and nonmenstrual migraine.

The aim of the present study was to investigate the effects of biofeedback training in the treatment of menstrual and nonmenstrual migraine. Accordingly, 39 female patients suffering from both migraine associated, and migraine not associated, with menstrual periods were drawn from a pool of research volunteers enrolled in a biofeedback treatment program for migraine headaches. All patients were required to complete 5 weeks of daily self-monitoring of headache and menstruation activity immediately before and after treatment, and again at 6-month follow-up. Within-subjects comparisons of the effects of biofeedback on menstrual and nonmenstrual migraine, and between-subjects comparisons of the effects of biofeedback on patients suffering predominantly from either menstrual or nonmenstrual migraine showed that biofeedback is just as effective in reducing menstrual migraine as it is in reducing nonmenstrual migraine. Questions as to whether or not these conclusions can apply to patients who experience migraine headaches only during, or shortly before or after, menstruation, are raised.

Metabolic engineering.

Metabolic engineering is the science that combines systematic analysis of metabolic and other pathways with molecular biological techniques to improve cellular properties by designing and implementing rational genetic modifications. As such, metabolic engineering deals with the measurement of metabolic fluxes and elucidation of their control as determinants of metabolic function and cell physiology. A novel aspect of metabolic engineering is that it departs from the traditional reductionist paradigm of cellular metabolism, taking instead a holistic view. In this sense, metabolic engineering is well suited as a framework for the analysis of genome-wide differential gene expression data, in combination with data on protein content and in vivo metabolic fluxes. The insights of the integrated view of metabolism generated by metabolic engineering will have profound implications in biotechnological applications, as well as in devising rational strategies for target selection for screening candidate drugs or designing gene therapies. In this article we review basic concepts of metabolic engineering and provide examples of applications in the production of primary and secondary metabolites, improving cellular properties, and biomedical engineering.