Degradation of mutant p53H175 protein by Zn(II) through autophagy.

TP53, one of the most important oncosuppressors, is frequently mutated in cancer. Several p53 mutant proteins escape proteolytic degradation and are highly expressed in an aberrant conformation often acquiring pro-oncogenic activities that promote tumor progression and resistance to therapy. Therefore, it has been vastly proposed that reactivation of wild-type (wt) function(s) from mutant p53 (mutp53) may have therapeutic significance. We have previously reported that Zn(II) restores a folded conformation from mutp53 misfolding, rescuing wild-type (wt) p53/DNA-binding and transcription activities. However, whether Zn(II) affects mutp53 stability has never been investigated. Here we show that a novel Zn(II) compound induced mutp53 (R175H) protein degradation through autophagy, the proteolytic machinery specifically devoted to clearing misfolded proteins. Accordingly, pharmacological or genetic inhibition of autophagy prevented Zn(II)-mediated mutp53H175 degradation as well as the ability of the Zn(II) compound to restore wtp53 DNA-binding and transcription activity from this mutant. By contrast, inhibition of the proteasome failed to do so, suggesting that autophagy is the main route for p53H175 degradation. Mechanistically, Zn(II) restored the wtp53 ability to induce the expression of the p53 target gene DRAM (damage-regulated autophagy modulator), a key regulator of autophagy, leading to autophagic induction. Accordingly, inhibition of wtp53 transactivation by pifithrin-α (PFT-α) impaired both autophagy and mutp53H175 degradation induced by curcumin-based zinc compound (Zn(II)-curc). Viewed together, our results uncover a novel mechanism employed by Zn(II)-curc to reactivate mutp53H175, which involves, at least in part, induction of mutp53 degradation via wtp53-mediated autophagy.

Stigmatization of schizophrenia as perceived by nurses, medical doctors, medical students and patients.

Stigmatization of schizophrenia is widespread and its genetic explanation may potentially increase the stigma. The present study investigated whether seeing schizophrenia as a genetic or environmental disorder might influence perceived beliefs towards people with schizophrenia and whether social stigmatizing attitudes were differently perceived the 202 subjects who were recruited. Perceived social stigmatizing attitudes were compared among participants who read two vignettes depicting a person with schizophrenia. Then, the Standardized Stigmatization Questionnaire (SSQ) was administered. A genetic explanation of schizophrenia was more frequently associated with stigmatizing attitudes. Also, there were higher levels of perceived stigmatization in medical students and medical doctors than in other groups based on their social experience or background. However, the sample size was small and this was a non-experimental design; also the SSQ would benefit from more cross-validation. About half of the participants perceived stigmatizing social attitudes. Finally, considering schizophrenia as a genetic disorder influenced participants perception of other people's beliefs about dangerousness and unpredictability and people's desire for social distance.

Factor-structure of the Italian version of the Scale Of Prodromal Symptoms (SOPS): a comparison with the English version.

AIMS: The 19-item 'Scale Of Prodromal Symptoms' (SOPS) and its semi-structured interview, the Structured Interview for Prodromal Symptoms (SIPS), have been developed to assess prodromes of psychosis. We assessed psychometric properties of the Italian version of the instrument. METHODS: We collected socio-demographic and clinical data of 128 people seeking first-time psychiatric help in a large Roman area, either as outpatients at community facilities or as inpatients in psychiatric wards of two general hospitals. Participants were administered the Italian version of the SOPS and the 24-item Brief Psychiatric Rating Scale (BPRS). Data were analysed through Pearson's correlation and factorial analysis. RESULTS: The English and Italian SOPS versions showed similar psychometric properties and factorial structure. The best-fit model was trifactorial, explaining 90% of total variance, and roughly corresponding to the positive, negative, and general dimensions, with disorganisation spreading over the other dimensions. Compared with the BPRS, the Italian version of the SOPS showed construct validity and convergent validity. CONCLUSIONS: The factor-structure of the Italian version of the SOPS is similar to those of the English and Spanish versions, in that the factors emerged are the same (positive, negative, and general symptoms). The scale could be used to assess at-risk people in early intervention services.

Influence of obstetric complication severity on brain morphology in schizophrenia: an MR study.

INTRODUCTION: The purpose of this study was to determine if a causal relationship exists between obstetric complications (OCs) severity and linear magnetic resonance (MR) measurements of brain atrophy in patients with schizophrenia. MATERIALS AND METHODS: Linear measurements of ventricular enlargement (bifrontal span, Evans ratio, and bicaudate ratio) and hippocampal atrophy (interuncal distance) were completed on MR images obtained in 47 patients with schizophrenia. Regression analysis was used to look at association with OCs severity, assessed by the "Midwife protocol" of Parnas and colleagues. The relationship between MR measurements and phenomenologic variables such as age at onset, illness duration, and exposure to antipsychotic medications was explored. The relationship between MR measurements, OCs severity, and symptom presentation was also investigated. RESULTS: OCs severity was significantly associated with MR measurements of ventricular enlargement (bifrontal span, Evans ratio). As the severity of OCs increased, bifrontal span and Evans ratio increased. This effect was independent of age at onset, illness duration, or even antipsychotic treatment. Interestingly, bifrontal span, Evans ratio, and OCs severity score all showed a significant positive correlation with hallucinatory symptomatology. CONCLUSION: Although confirmatory studies are needed, our findings would support the idea that environmental factors, in this case severe OCs, might partly contribute to ventricular abnormalities in schizophrenia.

Brain abnormalities in schizophrenia. A qualitative comparative study of schizophrenic patients and control individuals assessed by magnetic resonance imaging.

The present study examined, by means of Magnetic Resonance Imaging (MRI), the qualitative brain abnormalities in a group of 58 schizophrenic patients compared to a group of 58 matched control individuals. The possible relationships between these abnormalities and the demographic and clinical features of the participants in the study were also investigated. Schizophrenic patients presented a higher percentage of bland-moderate enlargement of the periencephalic-subarachnoid spaces (p=0.01) and a widespread cerebral atrophy, the latter below the threshold of significance (p=0.06). In the subset of patients with ventricular asymmetry (right larger than left) the age was significantly lower compared to the age of patients without this abnormality (p=0.04). In the subset of patients with cerebellar cisterns enlargement the age as well as the age of onset was higher in comparison to the one of patients without this abnormality (p=0.02; p=0.006). Taking together with previous studies, these findings underline the importance of qualitative assessment of brain morphology in research and clinical evaluation of patients with schizophrenia.

Intratester and intertester reliability of the cervical range of motion device.

The purpose of this study was to investigate the intratester and intertester reliability of the Cervical Range of Motion instrument (CROM) for measuring cervical flexion, extension, lateral flexion, and rotation. Twenty able-bodied subjects were tested by two testers on two different occasions. Pearson product-moment correlations for intratester reliability ranged from .63 to .90 for tester one and from .62 to .91 for tester two. Intertester reliability was good. Coefficients ranged from .80 to .87 for session one and .74 to .85 for session two. Paired data t-tests showed that there were no significant differences between testers or sessions (p = .01). The results suggest that the CROM has acceptable intratester and intertester reliability. The CROM has many benefits including ease of application and reliability. More research is needed on patients with cervical dysfunction.

Relationship between plasma somatomedin C and muscle performance in a geriatric male population.

The purpose of this study was to ascertain if a relationship existed between plasma somatomedin C (SmC) level, as an indicator of growth hormone secretion, and muscle performance. Eighteen community-dwelling men between the ages of 65 and 80 comprised the sample group. Muscle strength, power and endurance were measured isokinetically on the elbow flexors and extensors and on the knee flexors and extensors. No relationship was found between plasma SmC levels and measures of muscle performance (P greater than 0.05). The limitations of this study as well as suggestions for future studies were discussed. Further research is needed to assess the effects of growth hormone on muscle performance.

Hyposomatomedinemia in the men of a Veterans Administration Nursing Home: prevalence and correlates.

A previous study found hyposomatomedinemia to be common in the men of this VA Nursing Home. To gain information on the prevalence and correlates of this endocrine characteristic, we have measured plasma somatomedin C (SmC) in 69 Nursing Home men 55-95 years old (group I), and in 37 independent men 56-87 years old attending our geriatric medicine outpatient clinic (group II). In groups I and II, a clinical data base was compiled comprising: blood chemistries, measures of body composition - nutritional state and functional level, diagnoses, medications, and morbidity and mortality during the year after the SmC analysis. The mean +/- SD for SmC was significantly (p less than 0.05) lower in group I (0.35 +/- 0.21 units/ml) than in group II (0.45 +/- 0.13 units/ml). SmC less than 0.25 units/ml, a range consistent with severe growth hormone deficiency, was found almost exclusively in group I (31.8% of men in group I, 3.7% of men in group II). In either group I, or in groups I and II combined, SmC was significantly (p less than 0.05) correlated with body weight as percentage of ideal, midarm muscle circumference (MAMC) as percent of standard, diagnosis of cerebrovascular disease, and plasma testosterone level. Men with SmC values below 0.25 units/ml had significantly (p less than 0.05) lower values for body weight as percentage of ideal, and for MAMC as percentage of standard. Except for cerebrovascular disease, SmC did not correlate significantly (p greater than 0.05) with diagnosis, drugs, morbidity or mortality.

Insulin stimulates the dephosphorylation of phosphothreonine from fat-pad ATP-citrate lyase.

ATP-citrate lyase is phosphorylated in vivo at three amino acid residues on two peptide sequences (peptides a and b). Insulin action is known to increase the phosphorylation of peptide a. To study the effect of insulin on peptide b phosphorylation ATP-citrate lyase was radiolabeled in vivo by incubating fat pads with 32Pi. Following "cold chase", insulin action decreased the calculated specific radioactivity of peptide b to less than 30% of control whereas the specific radioactivity of peptide a increased 5-6 fold. The insulin induced decrease in peptide b phosphorylation was mainly due to a decrease in phosphothreonine phosphorylation. Isoproterenol treatment increased peptide a phosphorylation 4-6 fold but did not decrease peptide b phosphorylation. Specific radioactivity of ATP did not change significantly with hormone treatment. These results suggest that insulin action increases the dephosphorylation of peptide b by increasing the activity of a putative phosphothreonine phosphatase.

ATP-citrate lyase phosphorylation in rat adipose tissue.

Previous in vitro studies demonstrated that ATP-citrate lyase is phosphorylated by cyclic AMP-dependent protein kinase at peptide A, containing a phosphoserine residue, and by ATP-citrate lyase kinase at peptide B, containing both phosphoserine and phosphothreonine residues (Ramakrishna, S., Pucci, D. L., and Benjamin, W. B. (1983) J. Biol. Chem. 258, 4950-4956). In the present study, trypsin-digested, radiolabeled ATP-citrate lyase from rat epididymal fat pads was analyzed by high performance liquid chromatography. Phosphorylation occurred at three amino acid residues within two different peptide sequences; one (peptide a) contained phosphoserine and the other (peptide b) contained phosphoserine and phosphothreonine. The retention times and molecular weights were the same for peptides a and A and peptides b and B. Isoproterenol action increased peptide a phosphorylation and, to a lesser extent, peptide b phosphorylation. Insulin action also increased peptide a phosphorylation, but did not increase peptide b phosphorylation.

Dependence of ATP-citrate lyase kinase activity on the phosphorylation of ATP-citrate lyase by cyclic AMP-dependent protein kinase.

ATP-citrate lyase from rat liver and adipose tissue is phosphorylated by either ATP-citrate lyase kinase or catalytic subunit of cyclic AMP-dependent protein kinase to 0.5-0.6 mol/subunit. We previously demonstrated that the site phosphorylated by ATP-citrate lyase kinase (peptide B) is different from that phosphorylated by catalytic subunit of cyclic AMP-dependent protein kinase (peptide A) (Ramakrishna, S., Pucci, D. L., and Benjamin, W.B. (1981) J. Biol. Chem. 256, 10213-10216). ATP-citrate lyase phosphorylation by both protein kinases added simultaneously was increased synergistically. When ATP-citrate lyase was first phosphorylated by catalytic subunit of cyclic AMP-dependent protein kinase, the net phosphorylation of the fragments subsequently phosphorylated by lyase kinase increased about 6-fold. However, when ATP-citrate lyase was first phosphorylated by lyase kinase, there was no effect on the subsequent phosphorylation of the enzyme by cyclic AMP-dependent protein kinase. Alkaline phosphatase-dephosphorylated ATP-citrate lyase was phosphorylated by catalytic subunit of cyclic AMP-dependent protein kinase to 0.9-1.0 mol/subunit. However, dephospho-ATP-citrate lyase was not phosphorylated by lyase kinase. The addition of both protein kinases simultaneously phosphorylated ATP-citrate lyase up to 2 mol/subunit. Phosphorylation of dephospho-ATP-citrate lyase first by catalytic subunit of cyclic AMP-dependent protein kinase and ATP enabled the lyase to be phosphorylated by lyase kinase. Peptide mapping and phosphoamino acid analysis of dephospho-ATP-citrate lyase phosphorylated by catalytic subunit of cyclic AMP-dependent protein kinase and/or lyase kinase conclusively showed that phosphorylation of ATP-citrate lyase by ATP-citrate lyase kinase was completely dependent on peptide A phosphorylation by cyclic AMP-dependent protein kinase. Furthermore, increased phosphorylation when both protein kinases were added simultaneously was due to increased phosphorylation at peptide B.

ATP-citrate lyase kinase and cyclic AMP-dependent protein kinase phosphorylate different sites on ATP-citrate lyase.

ATP-citrate lyase was phosphorylated by highly purified cyclic AMP-independent protein kinase (ATP-citrate lyase kinase) or the catalytic subunit of cyclic AMP-dependent protein kinase. Each kinase phosphorylated ATP-citrate lyase equally but the combination of both kinases increased ATP-citrate lyase phosphorylation additively. When ATP-citrate lyase was phosphorylated with each kinase and partially digested with either L-1-tosylamido-2-phenylmethyl chloromethyl ketone-treated trypsin or Staphylococcus aureus protease followed by electrophoresis of the proteolytic products on sodium dodecyl sulfate-polyacrylamide gels or when the phosphorylated lyase was completely digested by these proteases followed by chromatography and electrophoresis, the results showed that the site phosphorylated by ATP-citrate lyase kinase was different from that phosphorylated by the catalytic subunit of cyclic AMP-dependent protein kinase. Only phosphoserine was found in lyase phosphorylated by the catalytic subunit of cyclic AMP-dependent protein kinase whereas phosphoserine and phosphothreonine were found in ATP-citrate lyase phosphorylated by lyase kinase.

Two-wavelength operation of a double-pass planar dye laser amplifier.

The simultaneous multiple line operation of a double pass dye laser amplifier is reported. Standard schemes have been used to generate two independently tunable lines and to scan one narrow line linearly around a fixed one. A simple, new scheme is illustrated, which allows symmetric splitting of narrow laser lines.