Cross-Tissue Exploration of Genetic and Epigenetic Effects on Brain Gray Matter in Schizophrenia.

Closely linking genetics and environment factors, epigenetics has been of increasing interest in psychiatric disease studies. In this work, we integrated single nucleotide polymorphisms (SNPs), DNA methylation of blood and saliva, and brain gray matter (GM) measures to explore the role of genetic and epigenetic variation to the brain structure changes in schizophrenia (SZ). By focusing on the reported SZ genetic risk regions, we applied a multi-stage multivariate analysis to a discovery dataset (92 SZ patients and 110 controls, blood) and an independent replication dataset (93 SZ patients and 99 controls, saliva). Two pairs of SNP-methylation components were significantly correlated (r = .48 and .35) in blood DNA, and replicated (r = .46 and .29) in saliva DNA, reflecting cross-tissue SNP cis-effects. In the discovery data, both SNP-related methylation components were also associated with one GM component primarily located in cerebellum, caudate, and thalamus. Additionally, another methylation component in NOSIP gene with significant SZ patient differences (P = .009), was associated with 8 GM components (7 with patient differences) including superior, middle, and inferior frontal gyri, superior, middle, and inferior temporal gyri, cerebellum, insula, cuneus, and lingual gyrus. Of these, 5 methylation-GM associations were replicated (P < .05). In contrast, no pairwise significant associations were observed between SNP and GM components. This study strongly supports that compared to genetic variation, epigenetics show broader and more significant associations with brain structure as well as diagnosis, which can be cross-tissue, and the potential in explaining the mechanism of genetic risks in SZ.

The MCIC collection: a shared repository of multi-modal, multi-site brain image data from a clinical investigation of schizophrenia.

Expertly collected, well-curated data sets consisting of comprehensive clinical characterization and raw structural, functional and diffusion-weighted DICOM images in schizophrenia patients and sex and age-matched controls are now accessible to the scientific community through an on-line data repository (coins.mrn.org). The Mental Illness and Neuroscience Discovery Institute, now the Mind Research Network (MRN, http://www.mrn.org/ ), comprised of investigators at the University of New Mexico, the University of Minnesota, Massachusetts General Hospital, and the University of Iowa, conducted a cross-sectional study to identify quantitative neuroimaging biomarkers of schizophrenia. Data acquisition across multiple sites permitted the integration and cross-validation of clinical, cognitive, morphometric, and functional neuroimaging results gathered from unique samples of schizophrenia patients and controls using a common protocol across sites. Particular effort was made to recruit patients early in the course of their illness, at the onset of their symptoms. There is a relatively even sampling of illness duration in chronic patients. This data repository will be useful to 1) scientists who can study schizophrenia by further analysis of this cohort and/or by pooling with other data; 2) computer scientists and software algorithm developers for testing and validating novel registration, segmentation, and other analysis software; and 3) educators in the fields of neuroimaging, medical image analysis and medical imaging informatics who need exemplar data sets for courses and workshops. Sharing provides the opportunity for independent replication of already published results from this data set and novel exploration. This manuscript describes the inclusion/exclusion criteria, imaging parameters and other information that will assist those wishing to use this data repository.

Three-way (N-way) fusion of brain imaging data based on mCCA+jICA and its application to discriminating schizophrenia.

Multimodal fusion is an effective approach to better understand brain diseases. However, most such instances have been limited to pair-wise fusion; because there are often more than two imaging modalities available per subject, there is a need for approaches that can combine multiple datasets optimally. In this paper, we extended our previous two-way fusion model called "multimodal CCA+joint ICA", to three or N-way fusion, that enables robust identification of correspondence among N data types and allows one to investigate the important question of whether certain disease risk factors are shared or distinct across multiple modalities. We compared "mCCA+jICA" with its alternatives in a 3-way fusion simulation and verified its advantages in both decomposition accuracy and modal linkage detection. We also applied it to real functional Magnetic Resonance Imaging (fMRI)-Diffusion Tensor Imaging (DTI) and structural MRI fusion to elucidate the abnormal architecture underlying schizophrenia (n=97) relative to healthy controls (n=116). Both modality-common and modality-unique abnormal regions were identified in schizophrenia. Specifically, the visual cortex in fMRI, the anterior thalamic radiation (ATR) and forceps minor in DTI, and the parietal lobule, cuneus and thalamus in sMRI were linked and discriminated between patients and controls. One fMRI component with regions of activity in motor cortex and superior temporal gyrus individually discriminated schizophrenia from controls. Finally, three components showed significant correlation with duration of illness (DOI), suggesting that lower gray matter volumes in parietal, frontal, and temporal lobes and cerebellum are associated with increased DOI, along with white matter disruption in ATR and cortico-spinal tracts. Findings suggest that the identified fractional anisotropy changes may relate to the corresponding functional/structural changes in the brain that are thought to play a role in the clinical expression of schizophrenia. The proposed "mCCA+jICA" method showed promise for elucidating the joint or coupled neuronal abnormalities underlying mental illnesses and improves our understanding of the disease process.

Cigarette smoking and white matter microstructure in schizophrenia.

The majority of patients with schizophrenia smoke cigarettes. Both nicotine use and schizophrenia have been associated with alterations in brain white matter microstructure as measured by diffusion tensor imaging (DTI). The purpose of this study was to examine fractional anisotropy (FA) in smoking and non-smoking patients with schizophrenia and in healthy volunteers. A total of 43 patients (28 smoking and 15 non-smoking) with schizophrenia and 40 healthy, non-smoking participants underwent DTI. Mean FA was calculated in four global regions of interest (ROIs) (whole brain, cerebellum, brainstem, and total cortical) as well as in four regional ROIs (frontal, temporal, parietal and occipital lobes). The non-smoking patient group had a significantly higher intellectual quotient (IQ) compared with the patients who smoked, and our results varied according to whether IQ was included as a covariate. Without IQ correction, significant between-group effects for FA were found in four ROIs: total brain, total cortical, frontal lobe and the occipital lobe. In all cases the FA was lower among the smoking patient group, and highest in the control group. Smoking patients differed significantly from non-smoking patients in the frontal lobe ROI. However, these differences were no longer significant after IQ correction. FA differences between non-smoking patients and controls were not significant. Among smoking and non-smoking patients with schizophrenia but not healthy controls, FA was correlated with IQ. In conclusion, group effects of smoking on FA in schizophrenia might be mediated by IQ. Further, low FA in specific brain areas may be a neural marker for complex pathophysiology and risk for diverse problems such as schizophrenia, low IQ, and nicotine addiction.

Voxel-based morphometric multisite collaborative study on schizophrenia.

Regional gray matter (GM) abnormalities are well known to exist in patients with chronic schizophrenia. Voxel-based morphometry (VBM) has been previously used on structural magnetic resonance images (MRI) data to characterize these abnormalities. Two multisite schizophrenia studies, the Functional Biomedical Informatics Research Network and the Mind Clinical Imaging Consortium, which include 9 data collection sites, are evaluating the efficacy of pooling structural imaging data across imaging centers. Such a pooling of data could yield the increased statistical power needed to elucidate effects that may not be seen with smaller samples. VBM analyses were performed to evaluate the consistency of patient versus control gray matter concentration (GMC) differences across the study sites, as well as the effects of combining multisite data. Integration of data from both studies yielded a large sample of 503 subjects, including 266 controls and 237 patients diagnosed with schizophrenia, schizoaffective or schizophreniform disorder. The data were analyzed using the combined sample, as well as analyzing each of the 2 multisite studies separately. A consistent pattern of reduced relative GMC in schizophrenia patients compared with controls was found across all study sites. Imaging center-specific effects were evaluated using a region of interest analysis. Overall, the findings support the use of VBM in combined multisite studies. This analysis of schizophrenics and controls from around the United States provides continued supporting evidence for GM deficits in the temporal lobes, anterior cingulate, and frontal regions in patients with schizophrenia spectrum disorders.

MTHFR 677C --> T genotype disrupts prefrontal function in schizophrenia through an interaction with COMT 158Val --> Met.

Understanding how risk genes cumulatively impair brain function in schizophrenia could provide critical insights into its pathophysiology. Working memory impairment in schizophrenia has been associated with abnormal dopamine signaling in the prefrontal cortex, which is likely under complex genetic control. The catechol-O-methyltransferase (COMT) 158Val --> Met polymorphism (rs4680), which affects the availability of prefrontal dopamine signaling, consistently stratifies prefrontal activation during working memory performance. However, the low-dopamine COMT 158Val allele does not confer increased risk for schizophrenia, and its effects on prefrontal function are not specific to the disorder. In the setting of other genetic variants influencing prefrontal dopamine signaling, COMT 158Val --> Met genotype may exert disease-specific effects. A second polymorphism, methylenetetrahydrofolate reductase (MTHFR) 677C --> T (rs1801133), has been associated with overall schizophrenia risk and executive function impairment in patients, and may influence dopamine signaling through mechanisms upstream of COMT effects. We found that the hypofunctional 677T variant was associated with decreased working memory load-dependent activation in the prefrontal and insular cortices in 79 schizophrenia patients, but not in 75 demographically matched healthy controls. Further, significant MTHFR x COMT genotype interactions were observed, which differed by diagnostic group: Reduced prefrontal activation was associated with the 677T and 158Val alleles in patients, but with 677C/C and 158Met/Met genotype in controls. These findings are consistent with epistatic effects of the COMT and MTHFR polymorphisms on prefrontal dopamine signaling, and suggest that in schizophrenia patients, the MTHFR 677T allele exacerbates prefrontal dopamine deficiency. The findings also suggest the importance of weighing COMT effects on prefrontal function within the context of MTHFR genotype.

Validation of the hospital anxiety and depression scale and the psychological disorder among premature ejaculation subjects.

Premature ejaculation (PE) is a common sexual dysfunction among the general population. PE has often been associated with a psychological state of mind. Hospital Anxiety and Depression Scale (HADS) can be used as an instrument to assess the emotional and psychological state. The present study was designed to assess the reliability and validity of the HADS in a Malaysian population. The validity and reliability were studied in subjects with and without PE. Test-retest methodology was used to assess the reliability whereas Cronbach's alpha was used to assess the internal consistency. In the control and the PE groups, the internal consistency was good and a high degree of internal consistency was observed for all 14 items. In the control group, the Cronbach's alpha values at baseline were from 0.811 to 0.834, whereas for retest, the Cronbach's alpha values were from 0.821-0.838 items. Intraclass correlation coefficient (ICC) was high for the control (0.797-0.868: baseline and 0.805-0.872: retest) and PE group (0.822-0.906: baseline and 0.785-0.887: retest). The high value of ICC and the internal consistency was due to high reliability and consistency of the items at 2-week interval. A degree of significance between the baseline and week-2 scores was observed across all items in the PE group but not in the control group. The HADS is a suitable, reliable, valid and sensitive instrument to measure the clinical change for anxiety and depression in the Malaysian population.

P53 protein expression in transitional cell carcinoma of the bladder - experience of the University of Malaya Medical Centre.

OBJECTIVES: To study the incidence of p53 oncoprotein overexpression and its relationship to tumour grade, stage and clinical prognosis in a cohort of local Malaysian patients. METHODS: All cases of transitional cell carcinoma (TCC) of the bladder diagnosed and treated at the University of Malaya Medical Centre from January 1995 to December 2000 were retrieved from the hospital records. Sections from paraffin-embedded tissues were retrieved and stained for p53 oncoprotein using immunohistochemistry techniques. P53 oncoprotein results were analyzed in relation to tumour grade, stage and clinical prognosis. Fisher's exact test was used to evaluate the relationship between categorical variables and the Kaplan-Meier procedure was used to assess survival outcomes. The Cox regression model was used for multivariate analysis. RESULTS: A total of 64 cases were studied. The mean follow-up period was 23.7 months. The number of p53 positive cases was significantly higher in high-grade (G3) (p = 0.006) and muscle-invasive tumours ( summation operator T2, p = 0.035). The status of p53 expression had no significant association with recurrence-free (p = 0.594) or overall survival (p = 0.955). In multivariate analysis, a multiplicity of tumours at presentation (p = 0.004) and a history of cigarette smoking (p = 0.016) were independent predictors of recurrence. Tumour stage (p = 0.024) was the single independent predictor for poor overall survival. CONCLUSIONS: Overexpression of p53 is associated with TCC of higher grade and tumour stage. It had no significant impact on prognosis in this cohort of TCC cases.

Alternate site testing for HbA1c using the Primus CLC330 GHb analyzer.

OBJECTIVE: To determine whether the Primus high-pressure liquid chromatography (HPLC) is suited to alternate site testing (AST) for HbA1c in a hospital diabetes outpatient clinic. RESEARCH DESIGN AND METHODS: Patients were attending the clinic for routine management of their diabetes. A number of diabetic patients with uremia (n = 11) were also investigated. HbA1c levels were measured in the outpatient setting by the Primus HPLC and in a more limited study the DCA-2000 instrument using the new 6-min assay cartridge. HbA1c measurements were also performed with Pierce affinity minicolumns and a Bio-Rad Variant HPLC. RESULTS: The Primus HPLC assay had low imprecision of 2.3, 1.6, and 1.0% for HbA1c levels of 4.7, 7.3, and 11.1%, respectively, and was not prone to interference by carbamylated hemoglobin as found for the ion-exchange Variant HPLC method. Method comparison studies showed that the bias and proportional error between the Pierce affinity minicolumn procedure (standardized with respect to an external quality control program) and the Primus HPLC (Y) was -0.4 and 1.2% respectively (n = 32). Similarly the bias and proportional error between the Primus and DCA-2000 methods was 0.7 and -2.5%. The Primus was shown to give falsely elevated HbA1c concentrations if the time between sequential injections was > 28 min. CONCLUSIONS: The Primus HPLC has a decided advantage over specialty AST instruments, like the DCA-2000, in not only meeting AST requirements but also allowing rapid automated batch processing of all laboratory HbA1c samples.

Sugar and fat, a recipe for disaster. New guidelines for the pharmacological management of diabetic dyslipidaemia.

Australia is embarking upon a more sophisticated approach to lipid-lowering, which is of particular relevance to those with diabetes. Diabetic dyslipidaemia is common and is usually managed in general practice. A step approach focusing on case finding, confirmation of diagnosis, lifestyle modification and pharmacological intervention should be cost effective in this high risk population (Figure 4). Although the focus of this review has been on diabetic dyslipidaemia, management should always consider overall cardiovascular risk.

Quantitative multivariate analysis of myoepithelioma and myoepithelial carcinoma.

The nuclear DNA content and morphologic data of 30 surgical samples (eight normal parotid gland, 12 myoepithelioma, and 10 myoepithelial carcinoma) were analyzed by the IBAS II analysis system. The results suggest that there are relationships between tumor ploidy distribution and prognosis, and that correct pathologic diagnosis can be obtained by the IBAS image system.

Metabolic approaches to the subclassification of polycystic ovary syndrome.

OBJECTIVES: To examine the relationship between various hormonal and metabolic variables in a large group of women with unequivocal evidence of polycystic ovarian syndrome (PCOS) to dissect out the metabolic heterogeneity of this condition. DESIGN: Cross-sectional observational study of PCOS (n = 122) and non-PCOS (n = 26) subjects. SETTING: Reproductive medicine unit in a tertiary teaching hospital. PATIENTS: Subjects with presumed PCOS were recruited from the Reproductive Medicine and Gynaecological Clinics and later confirmed as PCOS with recognized criteria. Several other subjects were identified through recruiting reference subjects. The PCOS population consisted of 122 patients. Reference subjects were recruited from partners of male factor infertility patients in the clinics and from the general population (n = 27). INTERVENTIONS: A 75 g 2-hour oral glucose tolerance test was performed on all subjects in their midluteal phase. Blood was taken at fasting and at 30, 60, 90, and 120 minutes. MAIN OUTCOME MEASURES: Age, body mass index (BMI), waist to hip ratio, levels of integrated glucose and insulin, concentrations of maximum insulin, sex hormone-binding globulin, T, triglyceride, apolipoproteins (Apo A1, B), high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol (LDLC). RESULTS: Five clusters could be identified. They are characterized as a nonobese group, a moderately obese group, and three very obese groups. The nonobese group (n = 41, BMI = 24.1) exhibited the lowest level of integrated insulin (236.4 mIU/L or microU/mL) and concentration of serum T (5.5 nmol/L). The moderately obese group had the second lowest level of integrated insulin (497.1 mIU/L) whereas the three very obese groups (n = 15, 13, and 5, respectively) had significantly higher but different levels of integrated insulin (group 3: 850.8 mIU/L; group 4: 1,131.5 mIU/L; and group 5: 1,531.9 mIU/L), triglyceride (group 3: 1.39 mmol/L; group 4: 1.76 mmol/L; and group 5: 2.78 mmol/L [1 mmol/L = 88mg/mL]), Apo B (group 3: 1.18 g/L; group 4: 1.08 g/L; and group 5: 1.55 g/L) and LDLC (group 3: 3.81 mmol/L; group 4: 3.05 mmol/L; and group 5: 5.06 mmol/L [1 mmol/L = 38.6 mg/100 mL]). CONCLUSIONS: The metabolic heterogeneity of the PCOS population is reflected at least partly in patients' levels of insulin, lipids, and lipoproteins, dependent and independent of BMI.

A sensitive method of screening for urinary porphobilinogen.

In this screening method for urinary porphobilinogen (PBG), urine is added to Dowex 2 resin under alkaline conditions in a test tube and mixed. The supernate is removed and the adsorbed PBG is eluted with acid and reacted with Ehrlich's reagent. We compared results with those by the Watson-Schwartz screening method, using urine samples from normal people with and without added PBG. At a PBG concentration of about five times the upper limit of normal, the resin method gave a sensitivity of 100%; the Watson-Schwartz method gave a sensitivity of 51%. At lower PBG concentrations of just over and twice the upper limit of normal, the sensitivity by the resin method was respectively 97% and 100%. With normal urine samples, the resin method gave negative results for all samples (100% specificity) and the Watson-Schwartz had 95% specificity. Our data indicate that the resin method is sensitive, specific, and reliable and is superior to the Watson-Schwartz method.

Ward and laboratory assessment of two second-generation blood-glucose monitoring systems.

We present an assessment of two blood-glucose monitoring systems--the Ames Glucostix reagent strips read on the Glucometer II and the Boehringer-Mannheim BM-Test-Glycemie 20-800 test strips read on the Reflolux II meter. Many nurses in the wards and a technologist in the laboratory assessed the systems. All operators achieved good precision with both systems at various glucose levels, the coefficients of variation for the technologist being less than 8% and those for the nurses being less than 11%. The Boehringer-Mannheim system gave good agreement with the laboratory values, with a mean bias of +0.14 mmol/L and limits of agreement between -1.38 mmol/L and +1.66 mmol/L. The Ames system gave poorer agreement, with a bias of +0.80 mmol/L and limits of agreement between -2.12 mmol/L and +3.72 mmol/L. Our study shows that both systems are capable of providing clinically-acceptable results when used by trained personnel. Although it took longer to use, the operators unanimously preferred the Boehringer-Mannheim system because generally it was easier to handle. This is due to a smaller amount of blood being needed which is easier to apply and to remove from the strip, and the cleaning material is not as likely to be left on the strip. Also, strip insertion is easier and there is sufficient time for this after the removal of blood.

Familial dysalbuminaemic hyperthyroxinaemia and other causes of euthyroid hyperthyroxinaemia.

Clinical and biochemical studies on a family in which 3 members have familial dysalbuminaemic hyperthyroxinaemia (FDH) are presented. They were clinically euthyroid with elevated serum thyroxine (T4) and free T4 indices but normal free T4 by equilibrium dialysis and normal serum triiodothyronine (total and free). All thyroid function tests on the remaining family members were normal. The inheritance is consistent with autosomal dominance. Also presented are data on 4 unrelated patients with FDH and two patients with T4 autoantibodies. The methods for detecting FDH, T4 antibodies and other causes of euthyroid hyperthyroxinaemia are now freely available. Since these anomalies may be more common than previously supposed, clinical awareness of the conditions is necessary to protect patients from the consequences of incorrect diagnosis of thyrotoxicosis.

Thyroid, renal, and hepatic function tests following cholecystography with high-dose contrast agents.

Serum biochemical tests were observed for about three weeks following oral cholecystography with fractionated high doses (6 g) of iopanoic acid (Telepaque) or sodium ipodate (Biloptin) in 24 and 29 patients, respectively. Both agents produced similar effects. No significant changes were seen in renal or hepatic function except for a mild increase in bilirubin on day 22. Serum urate decreased 10% on day 4, but the change was not significant. On days 4 and 11, there were significant increases in thyroid-stimulating hormone, thyroxine and free thyroxine index, and a moderate fall in triiodothyronine. Reverse triiodothyronine increased sharply on day 4. The pattern of changes observed suggests that these contrasts interfere with the extrathyroidal deiodination of iodothyronines. The temporary rise in thyroxine and free thyroxine index exceeded reference ranges in about half of all subjects, but they remained clinically euthyroid. Thyroid function tests should be interpreted with caution within three weeks of cholecystography.

The effects of iopodate on the serum iodothyronine pattern in normal subjects.

The ingestion by normal subjects of 3 g of sodium iopodate, which is widely used in routine oral cholecystography, resulted in significant decreases of serum total and free T3 to a nadir on day 4 which averaged 43% and 40%, respectively, below initial mean values. Total and free rT3 increased markedly to a peak on day 3, 244% and 189%, respectively, above initial mean values. Total and free T4 and free T4 index rose to a maximum on day 4, but these changes were not statistically significant. A marked TSH increase was also seen, most evident on day 3. All these changes reverted to baseline values by day 14 at a time when serum total iodide was still markedly elevated. It is concluded that the changes observed after iopodate were not due to alterations in serum binding proteins nor to an effect on thyroid gland by the large iodine component of iopodate, but were consistent with an effect on the peripheral metabolism of T4. Difficulty in interpreting routine thyroid function tests may occur for up to 14 days after oral cholecystography with iopodate.