Altered serotonin 1A binding in major depression: a [carbonyl-C-11]WAY100635 positron emission tomography study.

BACKGROUND: Serotonin 1A receptors (5-HT(1A)) are implicated in the pathophysiology of major depressive disorder (MDD) and in the action of selective serotonin reuptake inhibitors (SSRI). SSRI desensitize 5-HT(1A) and down-regulate 5-HT transporters (5-HTT) with the latter persisting for weeks after discontinuation of SSRI. MDD subjects are more likely to be homozygous for the functional 5-HT(1A) G(-1019) allele of the promoter polymorphism and are postulated to have higher 5-HT(1A) than healthy volunteers (controls). We measure 5-HT(1A) in MDD, assess the effects of antidepressant exposure (AE), and examine the role of the C(-1019)G polymorphism. METHODS: Genotyped and determined 5-HT(1A) binding potential (BP) by positron emission tomography (PET) using [carbonyl-C-11]-WAY-100635 in 28 medication-free MDD subjects during a current major depressive episode and 43 controls. RESULTS: No difference in BP between controls and MDD subjects (p = .235). There was a difference in BP comparing the controls, antidepressant naive (AN) MDD subjects, and subjects with AE across all regions (p = .013). Post hoc testing reveals higher BP in AN compared to controls (p = .008) and to AE (p = .007). The GG genotype is overrepresented in MDD subjects (p = .059), and BP appears higher with the G allele. CONCLUSIONS: AN have higher 5-HT(1A) than controls and AE suggesting a model of depression characterized by an over expression of autoinhibitory somatodendritic 5-HT(1A) receptors, perhaps due to the higher expressing G allele, that may result in reduced terminal field 5-HT release. AE appears to have long-term effects on 5-HT(1A).     

Prospective evaluation of endoscopic ultrasonography in the diagnosis of biliary microlithiasis in patients with normal transabdominal ultrasonography

Prior investigators have proposed microlithiasis as a causative factor for occult gallbladder diseases. Endoscopic ultrasonography (EUS) is potentially far more sensitive than transabdominal ultrasonography (TUS) in visualizing small stones. The aim of this study was to investigate the role of endoscopic ultrasonography (EUS) in the diagnosis of microlithiasis in patients with upper abdominal pain and normal TUS. Thirty-five patients with biliary-type abdominal pain and normal TUS results were prospectively studied. All patients underwent radial EUS by means of a GF UM-20 echoendoscope (Olympus Optical, Tokyo, Japan). Of 35 patients, 33 were revealed to have gallbladder sludge or small stones, and 21 had CBD sludge or microlithiasis. Nine patients were not available for follow-up; of the remaining patients, 13 underwent combined endoscopic biliary sphincterotomy and cholecystectomy, 10 underwent cholecystectomy, and 3 underwent biliary sphincterotomy alone. In a postoperative follow-up at 9.2 months, 25 patients (96.2%) were symptom free. EUS is an important diagnostic tool in patients with unexplained biliary colic. Cholecystectomy with or without EUS is an effective treatment modality in these settings. Presented in part as a poster at the Forty-Fifth Annual Meeting of The Society for Surgery of the Alimentary Tract, New Orleans, Louisiana, May 15–19, 2004. This study was supported by local funds from Digestive Disease Research Center, Tehran University of Medical Sciences, and has had no external financial support.     

Assessment of transient left ventricular dilation ratio via 2-day dipyridamole Tc-99m sestamibi nongated myocardial perfusion imaging

Background  The definition of an abnormal transient ischemic dilation (TID) ratio may be different according to stress type, type of isotope, and imaging protocols. The aim of this study was to derive the normal threshold and assessment of the TID ratio via 2-day dipyridamode stress/rest technetium 99m sestamibi myocardial perfusion single photon emission computed tomography (MPS). Methods and Results  We performed 2-day dipyridamole stress/rest Tc-99m sestamibi MPS in 665 patients. The TID ratio was calculated automatically with the Emory Cardiac Toolbox. The upper limit of normal (1.19) for the TID ratio was derived from 131 patients with a low (<5%) likelihood of coronary artery disease as mean +2 SDs. Patients with complete or partial reversible defects or multivessel-type or left anterior descending artery (LAD) territory perfusion abnormalities had higher TID ratios than the other patients. These patients had a higher frequency of an abnormal TID ratio (>1.19) as well. Binary logistic regression analysis showed that ischemia and LAD territory perfusion abnormality were independent predictors of an abnormal TID ratio. Conclusion  The threshold for an abnormal TID ratio via 2-day post-dipyridamole stress/rest Tc-99m sestamibi MPS was greater than 1.19. By use of this protocol, TID is not uncommon and it is related to a greater amount of ischemic burden as well as multivessel-type or LAD territory perfusion abnormality.     

The use of three-dimensional nanostructures to instruct cells to produce extracellular matrix for regenerative medicine strategies

Synthetic polymers or naturally-derived extracellular matrix (ECM) proteins have been used to create tissue engineering scaffolds; however, the need for surface modification in order to achieve polymer biocompatibility and the lack of biomechanical strength of constructs built using proteins alone remain major limitations. To overcome these obstacles, we developed novel hybrid constructs composed of both strong biosynthetic materials and natural human ECM proteins. Taking advantage of the ability of cells to produce their own ECM, human foreskin fibroblasts were grown on silicon-based nanostructures exhibiting various surface topographies that significantly enhanced ECM protein production. After 4 weeks, cell-derived sheets were harvested and histology, immunochemistry, biochemistry and multiphoton imaging revealed the presence of collagens, tropoelastin, fibronectin and glycosaminoglycans. Following decellularization, purified sheet-derived ECM proteins were mixed with poly(?-caprolactone) to create fibrous scaffolds using electrospinning. These hybrid scaffolds exhibited excellent biomechanical properties with fiber and pore sizes that allowed attachment and migration of adipose tissue-derived stem cells. Our study represents an innovative approach to generate strong, non-cytotoxic scaffolds that could have broad applications in tissue regeneration strategies.     

Tumor necrosis factor modulates transcription of myelin basic protein gene through nuclear factor kappa B in a human oligodendroglioma cell line

Tumor necrosis factor-alpha (TNF-alpha) is a major mediator of inflammation and it is involved in many neurological disorders such as multiple sclerosis. Levels of TNF-alpha and lymphotoxin-alpha have been found elevated in plaques, bloods, and cerebral spinal fluids from multiple sclerosis patients. The expression of myelin basic protein (MBP), a major protein of the myelin sheath, is affected by cytokines secreted by activated immune cells. To determine the signal transduction pathway involving tumor necrosis factor's action in myelination and demyelination, we have cloned and analyzed cis-elements on promoters of the human and mouse MBP genes. There are two putative nuclear factors kappa-B (NF-kappaB) cis-elements on the human and one on the mouse gene promoter. In an electrophoretic mobility shift assay, all three NF-kappaB cis-elements showed binding to a protein, which was recognized by an antibody against NF-kappaB P65 component. The specificity of the binding was demonstrated in a competitive assay using NF-kappaB consensus oligonucleotides. A two base pair site-directed mutation on the mouse NF-kappaB cis-element abolished its binding activity. We created a DNA construct by linking the mouse MBP gene promoter containing the NF-kappaB cis-element to luciferase gene. Transfection of this construct into a human oligodendroglioma cell line showed TNF-alpha increased the transgene expression. Furthermore the mutation of NF-kappaB site abolished TNF-alpha -induction of the transgene. The data demonstrate that NF-kappaB is the mediator between tumor necrosis factor's action and MBP gene expression. Elucidating the molecular mechanisms underlying TNF-alpha regulation of MBP gene expression provides new scientific bases for the development of therapy against oligodendrocyte-specific and myelin-related disorders such as multiple sclerosis.     

Multi-detector size-exclusion chromatography: a novel approach to calibration eliminating lag time estimation

In order to obtain the correct calibration of molecular-weight sensitive detectors (such as on-line viscometers or light scattering detectors) in size-exclusion chromatography, it is necessary to account for the (molecular) non-uniformity of the calibration standards. This is true even in the case of the narrow molecular weight distribution standards typically commercially available for calibration. Methods of taking this non-uniformity into consideration are presented. For a multi-detector size-exclusion chromatography system calibrated in this way, independent calibration curves can be established for each detector. Thus, at a given hydrodynamic volume, corresponding signals from the different detectors can be obtained. This eliminates the need to estimate additional parameters such as interdetector volumes or lag times for matching signals from different detectors. The practicability of this approach in the analysis of (molecularly) non-uniform polymers is demonstrated by combining on-line viscometer and concentration signals (to obtain intrinsic viscosity versus molecular weight information) for two different polymers, viz. polystyrene and poly(methyl methacrylate). The resultant intrinsic viscosity versus molecular weight data is then compared to that predicted from the literature.