Speaking to a Wider Audience about the Positive Contributions of Psychiatry.

We live in a world that is increasingly complex, intense, and stressful. Most people, at some time or other in their lives, can make good use of psychiatry as they map their course and steer their way through it. While this holds true, there also exists a very disturbing trend. No other branch of medicine suffers a similar, constant criticism, scrutiny and quite often downright vehement protest. Even the service users, who have been greatly benefitted, choose to stay mum for fear of stigmatization that may follow if they admit to have undergone therapy. The onus lies on both, the service users and providers alike, to take the positive contributions of psychiatry to the masses at large. All of us, especially medical professionals, need to consider our own attitudes and awareness. The recognition that anyone will break down if mental stress is high enough should help free us from a 'them and us' attitude. Reading about people's own experience of mental illness can promote understanding: Examples include a successful actress and a prize-winning author. For mental health practitioners, enabling service users to influence service development is another strong anti-stigma move. A cognitive behavior therapy approach can help individuals overcome the stigma felt and also cope better with discrimination. Also, we need to stand up against mental health discrimination wherever it is encountered.

Effects of valproic acid on organic acid metabolism in children: a metabolic profiling study.

Young children are at increased risk for valproic acid (VPA) hepatotoxicity. Urinary organic acid profiles, as a surrogate of mitochondrial function, were obtained in children 1.9 to 17.3 years of age (n = 52) who were undergoing treatment with VPA for seizure disorders. Age-matched patients receiving treatment with carbamazepine (CBZ; n = 50) and healthy children not undergoing treatment (n = 22) served as controls. Age-related changes in organic acid profiles were observed in all three groups. Although the untreated and CBZ control groups were indistinguishable from each other with respect to the principal-component analysis (PCA) score plots of the subjects, a distinct boundary was apparent between the VPA and each of the control groups. Interindividual variability was observed in the VPA-induced alterations in endogenous pathways corresponding to branched-chain amino acid metabolism and oxidative stress. The data suggest that more detailed metabolomic analysis may provide novel insights into biological mechanisms and predictive biomarkers for children at highest risk for serious toxicity.

Changes in serum osteocalcin and bone-specific alkaline phosphatase are associated with bone pain in donors receiving granulocyte-colony-stimulating factor for peripheral blood stem and progenitor cell collection.

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF) has been used to increase the number of CD34+ peripheral blood stem and progenitor cells collected by apheresis for use in autologous or allogeneic progenitor cell transplantation. The most frequent side effect of G-CSF treatment is bone pain, which occurs in over 80 percent of healthy progenitor cell donors. STUDY DESIGN AND METHODS: The possible mechanism of bone pain was investigated by measuring serum levels of osteocalcin (OC), bone-specific alkaline phosphatase (BAP), acid phosphatase (ACP), and tartrate-resistant acid phosphatase (TRAP) in seven healthy progenitor cell donors treated with human recombinant G-CSF administered subcutaneously for 5 consecutive days. RESULTS: All seven patients experienced bone pain during the treatment period. Serum levels of OC, BAP, ACP, and TRAP were measured in blood samples drawn on Days 0, 4, 5, 6, and 14. Levels of BAP were increased (p<0.05) over baseline on Days 4, 5, and 6, while those of OC decreased on Days 4, 5, and 6 (p<0.05). No significant changes occurred in ACP or TRAP levels. OC and BAP are considered markers of bone formation (osteoblast activity), and they correlate in many patients with metabolic bone disorders. The pattern of increased BAP and decreased OC has been reported in patients with osteolytic bone metastases. CONCLUSION: G-CSF treatment in healthy stem and progenitor cell donors may affect osteoblastic activity, and this activity may be associated with bone pain.

Peritoneal penetration of amphotericin B lipid complex and fluconazole in a pediatric patient with fungal peritonitis.

Fungal peritonitis is a rare event in patients receiving peritoneal dialysis. This case report describes the blood and dialysate concentrations of fluconazole and amphotericin B following intravenous administration in a 5-month-old infant with Candida albicans peritonitis receiving continuous cyclic peritoneal dialysis. Fluconazole rapidly and efficiently penetrated the peritoneal fluid achieving concentrations that exceed the minimal inhibitory concentration (MIC) for most Candida species. In contrast, the amount of amphotericin B in the dialysate was below the limit of quantification despite measurable blood concentrations. This suggests that fluconazole represents a better choice for antifungal therapy because of its excellent peritoneal penetration.

Short-term and long-term variability of plasma homocysteine measurement.

Within-person and methodological variability of a given analyte are important elements in determining whether an individual has altered concentrations of that analyte. We report the short-term (1 month) within-person, between-person, and methodological variability of plasma homocysteine in 20 healthy participants from whom samples were drawn weekly for 4 weeks. The short-term between-person variance was high, whereas within-person and methodological variances were relatively very low, giving a high reliability coefficient (R) for homocysteine (R = 0.94). The long-term (30 months) reliability coefficient was 0.65, but was greatly influenced by an outlier (R = 0.82 with the outlier excluded). The data suggest that an individual's plasma homocysteine concentration is relatively constant over at least 1 month, and a single measurement characterizes the average concentration reasonably well.

Production, analysis, and characterization of reference materials for prostate-specific antigen.

OBJECTIVE: To produce a set of three reference materials that mimic sera from patients with prostate disorders in the prostate-specific antigen (PSA) concentration range important for clinical screening for prostate cancer (approximately 0.5, approximately 4.0, and approximately 10.0 ng/mL), to analyze these reference materials in a large number of clinical laboratories using a variety of commercially available methods, and to characterize the molecular forms of PSA in them. METHODS: Units of serum from healthy individuals and from patients with varying degrees of elevated PSA were pooled, lyophilized, and distributed along with conventionally prepared, semen-supplemented proficiency testing samples to laboratories participating in the College of American Pathologists Basic Ligand Survey. The reference material and one of the standard Survey samples were fractionated by Sephacryl S-200-HR gel filtration chromatography. RESULTS: The Abbott IMx, Hybritech Tandem-E, Hybritech Tandem-R, and Tosoh AIA-Pack all measured PSA in the reference material fairly equally (agreement within +/- 12%). In contrast, the Abbott IMx results in the semen-supplemented Survey specimens were as much as 1.8-fold higher than the other three assays. Characterization of the molecular forms showed the reference material was approximately 90% alpha 1-antichymotrypsin-bound PSA, whereas the semen-supplemented Survey specimens were approximately 40% alpha 1-antichymotrypsin-bound PSA, which largely explained the difference in assay recoveries. CONCLUSIONS: Semen-free materials containing only endogenous PSA much more closely mimic real clinical specimens and should prove useful in efforts to standardize clinical PSA assays.

Methodological variables during analysis of in vivo cerebellar cyclic GMP, an indirect marker of nitric oxide release.

In light of the recent recognition of the physiological significance of nitric oxide, there is considerable interest in the methodological variables that can confound the results of the cerebellar cGMP analysis from in vivo experiments. In this study, using male Swiss Webster mice, the effect of such methodological variables as 1) weight of the animals; 2) tissue extraction procedures used in radioimmunoassay for cGMP; and 3) the commercial source of the assay kit on, harmaline-, pentylenetetrazole- or SNAP-induced increase in cerebellar cGMP in vivo were evaluated. Results indicate that mice in the 15- to 19-g weight range are most sensitive and best suited for in vivo drug effects on cerebellar cGMP. Furthermore, for the extraction of cerebellar cGMP, use of ice-cold 0.5N hydrochloric acid and subsequent dilution of the sample in assay buffer is the simplest and fastest method. Present data also indicate that the source of the radioimmunoassay kit has a significant effect on the cerebellar cGMP results. Based on the present results, the protocol developed and the guidelines drawn are timely and of high practical significance for research in the area of pharmacology of nitric oxide.

Modulation of rate at which serotonin-induced contraction decays in guinea pig trachea.

Stimulation of the type 2 serotonin (5-HT2) receptor in guinea pig trachea with 5-HT results in a contraction that decays in the continued presence of 5-HT. The decay of the 5-HT contraction has been proposed to be dependent on 5-HT2 receptor activation and to reflect desensitization of the receptor. The characteristics of the decay of the 5-HT contraction may also be dependent on other properties of the tissue. The effects of modulation of biochemical pathways implicated in airway smooth muscle contraction on the 5-HT contraction in isolated guinea pig trachea were determined with the use of a kinetic approach we developed previously. Decay of the 5-HT contraction was inhibited by cooling, increased by forskolin, 3-isobutylmethyl-1-xanthine, and 8-bromoadenosine 3',5'-cyclic monophosphate, and unaffected by staurosporine, H-7, H-8, phorbol 12,13-dibutyrate, and by inhibitors of the three major pathways of arachidonic metabolism. The results suggest that decay of the 5-HT contraction in guinea pig trachea is dependent on both the receptor and the biochemical state of the tissue.

Effect of cocaine on macromolecular syntheses and cell proliferation in cultured glial cells.

In the present study the effect of cocaine on thymidine, uridine and leucine incorporation was assessed in primary cortical glial and C6 glioma cells. Cocaine exposure for 24 h inhibited thymidine and uridine incorporation in cortical glial and C6 glioma cells. However, the effect of cocaine on uridine incorporation was less prominent compared to thymidine incorporation. High concentrations of cocaine inhibited leucine incorporation in C6 glioma cells but not in cortical glia. Cocaine exposure for four days decreased cell proliferation of cortical glial and C6 glioma cells. Cocaine-induced attenuation of macromolecular syntheses was not due to cell death since cocaine-treated cells were not stained with Trypan Blue and did not release lactate dehydrogenase into culture supernatants. Furthermore, cocaine had no effect on glutamate uptake either in cortical glia or in C6 glioma cells. These results indicate that cocaine inhibits macromolecular syntheses in glial cells. The inhibition of macromolecular syntheses in glial cells may be the mechanism involved in cocaine-induced fetal brain growth retardation.

Chronic administration of a nitric oxide synthase inhibitor, N omega-nitro-L-arginine, and drug-induced increase in cerebellar cyclic GMP in vivo.

N omega-nitro-L-arginine (NG-nitro-L-arginine) is a potent nitric oxide synthase inhibitor which crosses the blood brain barrier and does not undergo extensive metabolism in vivo. In this study, effect of chronic pretreatment of N omega-nitro-L-arginine (75 mg/kg, i.p., twice daily for 7 days) on the harmaline- (100 mg/kg, s.c.), picrotoxin- (4 mg/kg, s.c.), pentylenetetrazole- (50 mg/kg, i.p.), and L-glutamic acid- (400 micrograms/10 microliters/mouse, i.c.v.) induced increase in cerebellar cGMP was assessed. All the four drugs produced significant increase in cerebellar cGMP in vehicle pretreated control animals. Cerebellar cGMP increased induced by harmaline, picrotoxin, and L-glutamic acid was attenuated in N omega-nitro-L-arginine pretreated animals. These results indicate that in vivo cerebellar cGMP levels are increased by the prototype excitatory amino acid receptor agonist, L-glutamic acid and also by the drugs which augment the excitatory amino acid transmission. Furthermore, parenteral chronic administration of N omega-nitro-L-arginine blocks NO synthase in the brain and hence cerebellar cGMP response in chronic N omega-nitro-L-arginine treated animals could be used as a tool to assess the physiological functions of nitric oxide in vivo.

Mechanisms of nitrosothiol-induced antimitogenesis in aortic smooth muscle cells.

The purpose of this work was to investigate the effect of a nitrosothiol vasodilator, S-nitroso-N-acetylpenicillamine (SNAP), on serum-induced cell proliferation, thymidine uptake, RNA synthesis, protein synthesis and thymidine kinase activity in a cultured rat aortic smooth muscle cell line. SNAP decreased the rate of serum-stimulated cell proliferation, thymidine uptake and incorporation, uridine and leucine incorporation and thymidine kinase activity in concentration-dependent fashion. The threshold concentration of SNAP for inhibition of cell proliferation and thymidine uptake was similar and in the range of 1-3 microM. Uridine incorporation, indicative of RNA synthesis, was inhibited beginning at 10 microM SNAP, whereas leucine incorporation, indicative of protein synthesis, and thymidine kinase activity, an enzyme of importance to DNA synthesis, were inhibited beginning at 100 microM SNAP. These results indicate that inhibition of cell proliferation induced by relatively low concentrations of SNAP (1-10 microM) is independent of inhibition of protein synthesis or thymidine kinase activity, whereas higher concentrations of SNAP may inhibit cell proliferation by decreasing protein synthesis.

Effect of nitric oxide on mitogenesis and proliferation of cerebellar glial cells.

In the brain, nitric oxide (NO) has been identified as a messenger molecule and a mediator of excitatory amino acid-induced neurotoxicity. In this study, the effects of NO on serum-induced mitogenesis and cell proliferation of the cerebellar glial cells were assessed. NO-generating agent, S-nitroso-N-acetylpenicillamine (SNAP) increased intracellular cyclic guanosine monophosphate (cGMP) levels. Furthermore, 2 chemically dissimilar NO-generating agents, SNAP and sodium nitroprusside (SNP) inhibited serum-induced thymidine incorporation and cell proliferation. The antimitogenic effect of NO was mimicked by 8-bromo-cGMP and blocked by hemoglobin, a known inhibitor of NO. The effect of NO was not cytotoxic, since the cells were not stained with Trypan blue and did not show increased release of lactate dehydrogenase in the culture supernatants. However, NO-treated cells showed decreased conversion of tetrazolium to blue formazan suggesting that NO inhibited mitochondrial activity in the glial cells. These results demonstrate that NO inhibits serum-induced mitogenesis and cell proliferation of cultured rat cerebellar glial cells.

Nitric oxide decreases cytosolic free calcium in Balb/c 3T3 fibroblasts by a cyclic GMP-independent mechanism.

The purpose of this study was to investigate the effects of NO on cytosolic calcium levels in Balb/c 3T3 fibroblasts that were previously shown to lack soluble guanylate cyclase activity. Authentic NO as well as two NO-generating vasodilators, S-nitroso-N-acetyl-penicillamine and isosorbide dinitrate, decreased cytosolic calcium in these fibroblasts. The effect of NO and S-nitroso-N-acetylpenicillamine was concentration-dependent and, for the most part, reversible. Since S-nitroso-N-acetylpenicillamine did not increase either cGMP or cAMP, NO did not increase cGMP, and 8-bromo-cGMP did not alter cytosolic free calcium, we conclude that NO decreases cytosolic free calcium by a cyclic nucleotide-independent mechanism in Balb/c 3T3 fibroblasts.

Nitric oxide-generating vasodilators inhibit mitogenesis and proliferation of BALB/C 3T3 fibroblasts by a cyclic GMP-independent mechanism.

The purpose of this study was to investigate the effects of nitric oxide-generating vasodilators and 8-bromo-cGMP on serum-induced mitogenesis in BALB/c 3T3 fibroblasts that lack soluble guanylate cyclase activity. Two such vasodilators, S-nitroso-N-acetylpenicillamine and isosorbide dinitrate, decreased the incorporation of (3H)thymidine in these cells dose-dependently whereas 8-bromo-cGMP was ineffective at concentrations of up to 10 mM. Moreover, S-nitroso-N-acetylpenicillamine also inhibited cell proliferation, consistent with the data on (3H)thymidine incorporation. S-nitroso-N-acetylpenicillamine had no effect on cGMP accumulation, confirming previous studies that these cells lack soluble guanylate cyclase activity. Hemoglobin and FeSO4/ascorbate, agents that inhibit the actions of nitric oxide, both decreased S-nitroso-N-acetylpenicillamine-induced antimitogenesis, supporting the view that this effect was related to the generation of nitric oxide. The antimitogenic activity of S-nitroso-N-acetylpenicillamine was unlikely to be the expression of nitric oxide-induced degradation of serum mitogens, as indicated by the decrease of the antimitogenic activity on prolonged preincubation of SNAP in serum-containing medium. We conclude that nitric oxide-generating vasodilators inhibit serum-induced mitogenesis and cell proliferation in BALB/c 3T3 fibroblasts by a cGMP-independent mechanism.

Differences between the mechanisms of action of heat-stable and heat-labile enterotoxins of Escherichia coli.

The mucosal-to-serosal and serosal-to-mucosal fluxes of Na+ and Cl- were measured in control mice and mice treated with heat-stable (ST) and heat-labile (LT) enterotoxins in the presence or absence of: Ca2(+)-ionophore A23187, an activator of Ca2(+)-calmodulin; or phorbol-12-myristate-13-acetate (PMA), an activator of protein kinase C(PKC); or 1-(5-isoquinolinyl sulphonyl)-2-methyl piperazine (H-7), an inhibitor of PKC. There was net secretion of Na+ and CL- in both experimental groups in contrast to net absorption in the control group. The addition of ionophore or PMA or ionophore + PMA resulted in net secretion of Na+ and Cl- in the control group and the effect of ionophore and pMA was found to be additive. The addition of ionophore did not cause any change in electrolyte fluxes in the ST toxin treated group, however, it increased the net secretion of Na+ and Cl- in the LT toxin treated group. PMA increased the net secretion of Na+ and Cl- in the St toxin treated group, however, it did not cause any change in Na+ and Cl- fluxes in the LT toxin treated group. H-7 did not reverse the effect of ST toxin, however, it reversed the effect of LT toxin.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of Ca2(+)-calmodulin and protein kinase C in the secretory action of heat-labile enterotoxin of Escherichia coli in mice.

The mucosal-to-serosal and serosal-to-mucosal fluxes of Na+ and Cl- were carried out in control and heat-labile enterotoxin treated mice in the presence or absence of Ca2(+)-ionophore A23187, the activator of Ca2(+)-calmodulin or Phorbol-12-myristate-13-acetate (PMA), the activator of Protein kinase C (PKC) or 1-(5-isoquinolinyl sulphonyl)-2-methyl piperazine (H-7), an inhibitor of PKC. There was net secretion of Na+ and Cl- in experimental group in comparison to net absorption in control group. The addition of ionophore or PMA resulted in net secretion of Na+ and Cl- in control group. In experimental group ionophore increased the net secretion of Na+ and Cl- while, PMA could not cause any change in Na+ and Cl- fluxes in experimental group. Calmodulin activity remained unaltered in heat-labile enterotoxin treated mice as compared to control. H-7, reversed the effects of PMA and heat-labile enterotoxin. These studies demonstrate that heat-labile enterotoxin primarily involves PKC in its action.