Lipocalin 2 as a potential systemic biomarker for central serous chorioretinopathy.

No systemic biomarker of Central Serous Chorioretinopathy (CSCR) has been identified. Lipocalin 2 (LCN2 or NGAL), alone or complexed with MMP-9 (NGAL/MMP-9), is increased in several retinal disorders. Serum levels of LCN2 and NGAL/MMP-9 were measured in CSCR patients (n = 147) with chronic (n = 76) or acute/recurrent disease (n = 71) and in age- and sex-matched healthy controls (n = 130). Samples with CRP > 5 mg/L, creatinine > 100 µmol/L, and/or urea > 7.5 mmol/L were excluded. Serum LCN2 was lower in CSCR patients than controls (81.4 ± 48.7 vs 107.3 ± 44.5 ng/ml, p < 0.0001), and lower in acute/recurrent CSCR than controls (p < 0.001) and chronic CSCR (p = 0.006). Serum NGAL/MMP-9 was lower in CSCR patients than controls (47.2 ± 40.7 vs 74.1 ± 42.6, p < 0.0001), and lower in acute/recurrent CSCR than controls (p < 0.001) and chronic CSCR (p = 0.002). A ROC curve showed that for LCN2 serum levels, the 80-ng/ml cutoff value allows to discriminate acute/recurrent CSCR from controls with 80.3% sensitivity and 75.8% specificity, and for NGAL/MMP-9 serum levels, a 38-ng/ml cutoff value allows to discriminate acute/recurrent CSCR from controls with 69.6% sensitivity and 80.3% specificity. In both acute and chronic CSCR, low serum LCN2 and NGAL/MMP-9, provide a biological link between the two CSCR forms, and potential susceptibility to oxidative stress and innate immune dysregulation in CSCR.

GSK3beta, a centre-staged kinase in neuropsychiatric disorders, modulates long term memory by inhibitory phosphorylation at serine-9.

Accumulating evidence implicates deregulation of GSK3ss as a converging pathological event in Alzheimer's disease and in neuropsychiatric disorders, including bipolar disorder and schizophrenia. Although these neurological disorders share cognitive dysfunction as a hallmark, the role of GSK3ss in learning and memory remains to be explored in depth. We here report increased phosphorylation of GSK3ss at Serine-9 following cognitive training in two different hippocampus dependent cognitive tasks, i.e. inhibitory avoidance and novel object recognition task. Conversely, transgenic mice expressing the phosphorylation defective mutant GSK3ss[S9A] show impaired memory in these tasks. Furthermore, GSK3ss[S9A] mice displayed impaired hippocampal L-LTP and facilitated LTD. Application of actinomycin, but not anisomycin, mimicked GSK3ss[S9A] induced defects in L-LTP, suggesting that transcriptional activation is affected. This was further supported by decreased expression of the immediate early gene c-Fos, a target gene of CREB. The combined data demonstrate a role for GSK3ss in long term memory formation, by inhibitory phosphorylation at Serine-9. The findings are fundamentally important and relevant in the search for therapeutic strategies in neurological disorders associated with cognitive impairment and deregulated GSK3ss signaling, including AD, bipolar disorder and schizophrenia.

Glycogen synthase kinase-3beta, or a link between amyloid and tau pathology?

Phosphorylation is the most common post-translational modification of cellular proteins, essential for most physiological functions. Deregulation of phosphorylation has been invoked in disease mechanisms, and the case of Alzheimer's disease (AD) is no exception: both in the amyloid pathology and in the tauopathy are kinases deeply implicated. The glycogen synthase kinase-3 (GSK-3) isozymes participate in diverse cellular processes and important signalling pathways and have been implicitly linked to diverse medical problems, i.e. from diabetes and cancer to mood disorders and schizophrenia, and in the neurodegeneration of AD. Here, we review specific aspects of GSK-3 isozymes in the framework of recent data that we obtained in novel transgenic mouse models that robustly recapitulate the pathology and mechanistical problems of AD.

Restorative impact of rabeprazole on gastric mucus and mucin production impairment during naproxen administration: its potential clinical significance.

Rabeprazole augments gastric mucus and mucin production in humans. However, its potential restorative impact on gastric mucus and mucin production impairment, resulting from administration of naproxen, remained to be explored. Therefore, we measured the content of mucus and mucin in gastric juice (GJ) before and after administration of naproxen with rabeprazole or placebo. The study was approved by HSC at KUMC and conducted in 21 asymptomatic, H. pylori-negative volunteers in a double-blind, placebo-controlled, crossover design. The content of gastric mucus in GJ, after exhaustive dialysis and complete lyophilization, was assessed gravimetrically, whereas the content of mucin was measured after its purification with equilibrium density-gradient ultracentrifugation in CsC1. Gastric mucus secretion during administration of naproxen with placebo declined significantly both in basal (by 44%; P < 0.001) and in pentagastrin-stimulated (by 35%; P < 0.001) conditions. Coadministration of rabeprazole significantly restored the naproxen-induced impairment in mucus production in basal conditions (by 47%; P < 0.01) and by 22% during stimulation with pentagastrin. Gastric mucin secretion during naproxen/placebo administration also declined significantly in both basal (by 39%; P < 0.01) and stimulated (by 49%; P = 0.003) conditions. Rabeprazole also significantly restored the naproxen-induced decline of gastric mucin output during pentagastrin-stimulated conditions (by 67%; P = 0.003) and by 40% in basal conditions (P = 0.05). The restorative capacity of rabeprazole on the quantitative impairment of gastric mucus and mucin during administration of naproxen may translate into a clinical benefit of protection of the upper alimentary tract from NSAID-related mucosal injury.

Estimation of sodium and potassium concentrations in the uterine fluid of mares by microdialysis and ion chromatography.

Equine blastocyst fluid has a lower [Na+], a higher [K+] and a lower osmolality than does normal blood serum. Based on the assumptions that the sodium pump is primarily responsible for fluid accumulation and that ions transported actively into the blastocyst increase blastocyst osmolality above that of the external medium, we hypothesized that the [Na+] and the osmolality of mare uterine fluid are lower than those of blastocyst fluid. Microdialysis and ion chromatography were used to estimate [Na+] and [K+] of uterine fluid. Mares (n=10) were used for in vivo measurements at different stages of the oestrous cycle and during early pregnancy. On the basis of the results of studies in vitro, the mean +/- SD intrauterine [Na+] and [K+] were 110.0 +/- 15.1 and 12.3 +/- 6.8 mmol l(-1), respectively. These results indicate that uterine fluid of mares has a lower [Na+] and a higher [K+] than does normal serum. However, the [Na+] in uterine fluid is considerably higher than that of blastocyst fluid. Thus, these results do not support our hypothesis and the mechanism of production of hypotonic blastocyst fluid in horses remains unclear.

Development of an observation method for assessing pain behaviors in children with juvenile rheumatoid arthritis.

OBJECTIVE: To develop an observation method for assessing pain behaviors in children with juvenile rheumatoid arthritis (JRA). METHODS: Thirty children with JRA performed a standardized sequence of activities for video recording, and correlations between the pain behaviors observed on the videotapes and established measures of pain, depression, and functional disability were determined. RESULTS: Pain behaviors were reliably observed (kappa coefficients 0.53-0.79). Total pain behaviors were significantly correlated with subjective reports of pain (r = 0.50) and disability levels (r = 0.64). These behaviors were not significantly associated with children's depression ratings (r = 23). CONCLUSION: The results indicate that the behavioral observation method provides a reliable and valid measure of pain associated with JRA. Measurement of pain behaviors may be especially useful in treatment outcome studies because these behaviors are relatively independent of depression.

Pediatric cancer: predicting sibling adjustment.

Within the context of a coping model, the present study attempted to determine variables associated with good and poor adjustment for siblings (N = 129) of pediatric cancer patients. Family cohesion and adaptability were found to mediate the impact of pediatric cancer on the healthy sibling. Specifically, high levels of family cohesion and adaptability were associated with better adaptation for siblings. Some support also was found for Folkman, Schaefer, and Lazarus' (1979) coping model, as the following variables, proposed by the model, predicted sibling adjustment to pediatric cancer: health/energy/morale, social support, and utilitarian resources. Results from this study stress the need to examine the child in a variety of ecological contexts (e.g., parental and familial functioning; neighborhood/community support).

Juvenile rheumatoid arthritis: pain-related and psychosocial aspects and their relevance for assessment and treatment.

Objective. To review literature in the area of juvenile rheumatoid arthritis that has focused on pain experience, functional losses, and psychosocial functioning. Methods. This article provides a critical review of research addressing these three primary issues. Results. Subjective and behavioral measures have been developed to assess pain in juvenile rheumatoid arthritis patients, but further work is needed to determine the validity and reliability of these instruments. Tools to assess functional losses in juvenile rheumatoid arthritis patients also appear promising in preliminary studies. Patients can be at risk for difficulties in psychosocial functioning, although research suggests that there are a variety of family, parental, and child variables that influence child and sibling adaptation. Methodologic shortcomings prevent definitive conclusions in this area. Conclusions. Although methodologic limitations have plagued this research in the past, new advances are facilitating improved understanding of children and adolescents with juvenile rheumatoid arthritis. Implications for future study with this challenging population are offered.

Clinical perspectives of some neuroleptics through development and application of their assays.

Attempts to investigate relationships between plasma levels of neuroleptics and therapeutic outcome in schizophrenic patients have been hampered due to such factors as the chemical nature of these drugs, their metabolism, and the very heterogeneous nature of the disease states. Two clinical studies are described that investigate the relationship between plasma levels of fluphenazine (FLU) and its metabolites and therapeutic outcome in schizophrenic patients. In the first of these studies the levels of FLU and fluphenazine sulfoxide (FLUSO) in schizophrenics receiving either 5 or 25 mg of fluphenazine decanoate (FLUD) intramuscularly every 2 weeks were monitored. Patients given 25 mg of FLUD required 3 months to reach plasma level steady state. The results suggest that such patients, when being switched from the oral to the depot formulation of FLU, should continue to receive oral supplementation during the 1st 3 months after conversion. The relationship between log-transformed plasma levels at 26 and 38 weeks with subsequent psychotic exacerbation was investigated with the use of logistic regression and survival analysis. Both demonstrated significant relationships between FLU plasma levels and a risk of psychotic exacerbation at 26 and 38 weeks. The possibility of any correlations between neurological side effects and plasma concentrations were also investigated, with statistically significant correlations between FLU levels and akinesia found at 2 and 26 weeks. In the second of these studies the levels of FLU, FLUSO, 7-hydroxyfluphenazine (7-OHFLU), and fluphenazine N4'(-)-oxide (FLUNO) in schizophrenics receiving 5, 10, or 20 mg of oral fluphenazine dihydrochloride daily for 4 weeks were monitored. The relationships between log-transformed plasma levels, disabling side effects, and global improvement were examined by logistic regression for the 4-week period. The study showed a significant correlation between increases in both plasma levels and disabling side effects such that at a plasma level of 2.7 ng/ml, approximately 90% of acutely ill patients experienced disabling side effects. Conversely, the study also showed that at a plasma level of 0.67 ng/ml, 48% of patients experienced improvement without the development of disabling side effects. When relationships between metabolite levels, disabling side effects, and global improvement were examined by logistic regression, a stronger correlation between disabling side effects and FLUNO levels than between side effects and FLU levels was found.(ABSTRACT TRUNCATED AT 400 WORDS)

Synthesis of the N-oxides of phenothiazine antipsychotic agents.

Chlorpromazine N-oxide, fluphenazine N4'-oxide, prochlorperazine N4'-oxide, sulforidazine N-oxide, and trifluoperazine N4'-oxide were synthesized by oxidation of the designated nitrogen atom in the N-10 side chain of the respective parent drug with 3-chloroperoxybenzoic acid. In the case of trifluoperazine, a stepwise increase in the amount of oxidant yielded the N1',N4'-dioxide and N1',N4',S-trioxide. The N',S-dioxides of chlorpromazine and sulforidazine were obtained by hydrogen peroxide oxidation of the appropriate parent drug.

Retrospective review of sexual and marital satisfaction of spinal cord injury and diabetic males post penile injection or implant.

This study compares sexual activity and sexual and marital satisfaction in spinal-cord-injured (SCI) and diabetic men who had received either a penile prosthesis or papaverine/phentolamine injections for the treatment of impotence. Questionnaires were used retrospectively to assess changes pre- and post-procedure in 30 SCI and 26 diabetic males. Regardless of whether they received a penile prosthesis or injections, both diabetic and SCI males reported increases in diversity and frequency of sexual activities and satisfaction with sexual activities post-implant/injection. When compared with a normative sample, both diabetic and SCI males were found to have poorer body image post-implant/injection. They also reported more disagreement with their partners but engaged in more activities together.

The metabolites of chlorpromazine N-oxide in rat bile.

1. The metabolism of chlorpromazine N-oxide was studied in female rats after a 20 mg/kg single i.p. dose. 2. Metabolites identified in urine and faeces were chlorpromazine, 7-hydroxychlorpromazine, chlorpromazine sulphoxide, N-desmethylchlorpromazine and N-desmethylchlorpromazine sulphoxide. As these same five metabolites were previously shown to be present after oral administration this indicates that reduction of chlorpromazine N-oxide occurs not only in the gastrointestinal tract but also at other sites. 3. The metabolism of chlorpromazine N-oxide was studied following its administration by either i.p., i.v. or oral routes to female rats in which the bile duct was cannulated. 4. There were no qualitative differences between the three routes of administration with respect to the metabolites identified. With the exception of the absence of N-desmethylchlorpromazine and N-desmethylchlorpromazine sulphoxide, all metabolites previously identified in urine and faeces were also present in bile. 5. Additionally there were three compounds present in rat bile which were not identified in urine or faeces. These were chlorpromazine N-oxide, chlorpromazine N,S-dioxide and 7-hydroxychlorpromazine O-glucuronide. This is the first unequivocal evidence for the identification of intact 7-hydroxychlorpromazine O-glucuronide in any species. 6. The inability to detect chlorpromazine N-oxide and chlorpromazine N,S-dioxide in the faeces of rats is likely to be due to the reduction of the N-oxide group on the passage of these biliary metabolites down the intestinal tract.

The effectiveness of different methods of defining traumatic brain injury in predicting postdischarge adjustment in a spinal cord injury population.

Traumatic brain injury (TBI) can occur concomitantly with spinal cord injury (SCI). Much of the initial work in this area has focused on identifying coincidence rates and risk factors; less has focused on possible long-term implications of TBI when it occurs with SCI. In this study, SCI/TBI and neurologically matched SCI-only groups were formed on the basis of clinicians' ratings of neuropsychologic test scores. SCI/TBI and SCI-only groups were also formed using Halstead Category cutoff scores, presence/absence of loss of consciousness, and clinicians' ratings of severity of TBI. Dependent measures assessed an average of two years postinjury measured personal, social, and family adjustment of the individual with SCI and that of a significant other. Loss of consciousness and nonconsensus clinical ratings of presence/absence of TBI predicted postdischarge adjustment poorly. Severity ratings in the moderate to severe range, and Category cutoff scores did have some predictive value, with patients defined as impaired being more likely to report adjustment difficulties or being described as having adjustment difficulties by a significant other. The difficulty of making unequivocal diagnoses of TBI in this population is discussed and implications for future research delineated.

Effect of bullet removal on subsequent pain in persons with spinal cord injury secondary to gunshot wound.

The prevention or minimization of future pain is often cited as a reason for removal of the bullet from patients who have incurred a spinal cord injury secondary to a gunshot wound. In an attempt to examine this assumption, multimodal pain ratings were recorded for 14 patients with spinal cord injury due to a gunshot wound in whom the bullet was still present, 14 neurologically matched patients with spinal cord injury due to a gunshot wound in whom the bullet was removed, and 28 control patients with spinal cord injury unrelated to a gunshot wound who were neurologically matched to the first two groups. The results suggest that persons who sustain a spinal cord injury secondary to gunshot wounds report more pain than those injured in other ways. In addition, there was no indication that surgical removal of the bullet was helpful in reducing subsequent pain either early in the rehabilitation process or at 1 year postinjury. The location of the bullet and the type of pain that subsequently developed were not correlated with the initial decision to surgically remove the bullet. Implications for further study and clinical practice are discussed.

The metabolism of chlorpromazine N-oxide in man and dog.

1. The metabolism of chlorpromazine N-oxide was studied in female dogs and adult male humans after a single oral dose. 2. There was extensive metabolism in both species in that between four and seven metabolites were separately identified in urine and faeces. Apart from chlorpromazine N-oxide, chlorpromazine N,S-dioxide was the only isolated metabolite which retained the N-oxide group. The other identified metabolites were chlorpromazine and its 7-hydroxy, sulphoxide, N-desmethyl, 7-hydroxy-N-desmethyl and N-desmethylsulphoxide derivatives. 3. With dog samples, metabolites were separated by h.p.l.c. and individually collected prior to mass spectrometric analysis. With human samples, metabolites were directly subjected to h.p.l.c.-mass spectrometric determination. With all metabolites their structures were confirmed by direct comparison of their mass spectra and chromatographic behaviours with those of authentic samples. 4. The metabolites identified in urine and faeces were for the most part the same in both species, with the exceptions that chlorpromazine N-oxide was identified in the faeces of dog only and 7-hydroxy-N-desmethylchlorpromazine was identified in the urine of man only. 5. The observation of N-oxide compounds in the excreta of both man and dog contrasted with that for the previously studied rat, where no such compounds were detected.

The metabolism of chlorpromazine N-oxide in the rat.

1. The metabolism of chlorpromazine N-oxide was studied in female rats after a 20 mg/kg single oral dose. 2. Metabolites identified in both urine and faeces were chlorpromazine, 7-hydroxychlorpromazine, chlorpromazine sulphoxide, N-desmethylchlorpromazine and N-desmethylchlorpromazine sulphoxide. 3. Metabolites were separated by h.p.l.c. or g.l.c. prior to mass spectrometric analysis. The structures of the metabolites were confirmed by direct comparison of their mass spectra and chromatographic behaviours with those of authentic compounds. 4. Chlorpromazine N-oxide and any metabolite which retained the intact N-oxide function, such as chlorpromazine, N,S-dioxide, could not be identified in any of the extracts. 5. When 3H-chlorpromazine N-oxide was administered under the same conditions; approximately twice as much radioactivity was excreted in the faeces (52.1 +/- 9.7%) as in the urine (26.9 +/- 7.2%).

Synthesis and properties of haptens for the development of radioimmunoassays for thioridazine, mesoridazine, and sulforidazine.

For the separate development of radioimmunoassay procedures for thioridazine and its two major active metabolites, mesoridazine and sulforidazine, three haptens, respectively, 2-methylthio-, 2-methylsulfinyl-, and 2-methylsulfonyl-substituted 10-[2-[1-(2-carboxyethyl)-2-piperidinyl]ethyl]-10H-phenothiazine, were synthesized and characterized. Thioridazine hapten was coupled to bovine serum albumin, whereas the haptens for mesoridazine and sulforidazine were coupled to porcine thyroglobulin. The number of hapten residues per mole of carrier protein was determined in each case by an ultraviolet spectrophotometric method. Polyclonal antibodies to each hapten-protein conjugate were obtained in rabbits, and titers of the antisera were checked by evaluating their binding characteristics to the appropriate tritiated analyte. A hapten for the ring sulfoxide metabolite of thioridazine was also synthesized.