Measurement of the alpha4beta2* nicotinic acetylcholine receptor ligand 2-[(18)F]Fluoro-A-85380 and its metabolites in human blood during PET investigation: a methodological study

2-[(18)F]fluoro-A-85380 (2-[(18)F]FA) is a new radioligand for noninvasive imaging of alpha4beta2* nicotinic acetylcholine receptors (nAChRs) by positron emission tomography (PET) in human brain. In most cases, quantification of 2-[(18)F]FA receptor binding involves measurement of free nonmetabolized radioligand concentration in blood. This requires an efficient and reliable method to separate radioactive metabolites from the parent compound. In the present study, three analytical methods, thin layer chromatography (TLC), high-performance liquid chromatography (HPLC) and solid phase extraction (SPE) have been tested. Reversed-phase TLC of deproteinized aqueous samples of plasma provides good estimates of 2-[(18)F]FA and its metabolites. However, because of the decreased radioactivity in plasma samples, this method can be used in humans over the first 2 h after radioligand injection only. Reliable quantification of the parent radioligand and its main metabolites was obtained using reversed-phase HPLC, followed by counting of eluted fractions in a well gamma counter. Three main and five minor metabolites of 2-[(18)F]FA were detected in human blood using this method. On average, the unchanged 2-[(18)F]FA fraction in plasma of healthy volunteers measured at 14, 60, 120, 240 and 420 min after radioligand injection was 87.3+/-2.2%, 74.4+/-3%, 68.8+/-5%, 62.3+/-8% and 61.0+/-8%, respectively. In patients with neurodegenerative disorders, the values corresponding to the three last time points were significantly lower. The fraction of nonmetabolized 2-[(18)F]FA in plasma determined using SPE did not differ significantly from that obtained by HPLC (+gamma counting) (n=73, r=.95). Since SPE is less time-consuming than HPLC and provides comparable results, we conclude that SPE appears to be the most suitable method for measurement of 2-[(18)F]FA parent fraction during PET investigations.     

Synovial cyst that compressed the peroneal nerve: case report

We report a case of synovial cyst that induced the compression of the peroneal nerve. The patient presented with foot drop and impossibility to stand on his right heel associated with numbness of anterolateral surfaces of the right shin and the dorsal surface of the foot within a 1-month period. Based on the clinical examination and electroneuromyography data, the compression of nerve by soft elastic mass was confirmed. We treated the patient by total microsurgical excision of the soft elastic mass adherent to the right peroneal nerve at the top third of the patient’s shin. Histological study revealed the cyst of synovial membrane. During the postoperative period, a force in extensors of the right foot was restored. On the seventh day, the patient began to step on his right heel, while the zone of tenderness and temperature hypoesthesia in the area of the peroneal nerve innervation remained.     

Frictional Behavior of Individual Vascular Smooth Muscle Cells Assessed By Lateral Force Microscopy

With the advancement of the field of biotribology, considerable interest has arisen in the study of cell and tissue frictional properties. From the perspective of medical device development, the frictional properties between a rigid surface and underlying cells and tissues are of a particular clinical interest. As with many bearing surfaces, it is likely that contact asperities exist at the size scale of single cells and below. Thus, a technique to measure cellular frictional properties directly would be beneficial from both a clinical and a basic science perspective. In the current study, an atomic force microscope (AFM) with a 5 μm diameter borosilicate spherical probe simulating endovascular metallic stent asperities was used to characterize the surface frictional properties of vascular smooth muscle cells (VSMCs) in contact with a metallic endovascular stent. Various treatments were used to alter cell structure, in order to better understand the cellular components and mechanisms responsible for governing frictional properties. The frictional coefficient of the probe on VSMCs was found to be approximately 0.06. This frictional coefficient was significantly affected by cellular crosslinking and cytoskeletal depolymerization agents. These results demonstrate that AFM-based lateral force microscopy is a valuable technique to assess the friction properties of individual single cells on the micro-scale.     

Recombinant Kv Channels at the Membrane of Escherichia coli Bind Specifically Agitoxin2

Potassium voltage-gated channels (Kv) are considered as molecular targets in a number of serious neuronal, immune, and cardiac disorders. Search for efficient low-molecular weight modulators of Kv channel function provides a basis for the development of an appropriate therapy for various Kv-mediated diseases. We report here on a new bacterial cell-based system, which is suitable for study of interactions between ligands and ligand-binding sites of eukaryotic Kv1.3 and Kv1.1 channels. To create this system, high-level expression of KcsA-Kv1.3 and KcsA-Kv1.1 hybrid proteins (ligand-binding sites of Kv1.3 or Kv1.1 fused with prokaryotic KcsA potassium channel) was achieved in the plasma membrane of Escherichia coli. An efficient procedure of E. coli conversion to intact spheroplasts was developed. We demonstrate that fluorescently labeled agitoxin 2 binds specifically to high-affinity and lower-affinity sites of KcsA-Kv1.3 and KcsA-Kv1.1, respectively, at the membrane of spheroplasts. Number of binding sites per cell is estimated to be (1.0 ± 0.6) ×10⁵ and (0.3 ± 0.2) ×10⁵ for KcsA-Kv1.3- and KcsA-Kv1.1-presenting cells, respectively, that allows reliable detection of ligand–receptor interactions by confocal laser scanning microscopy. This bacterial cell-based system is intended for screening of ligands to membrane-embedded pharmaceutical targets. The work was presented at Russian-American Conference on Nanobiology and Nanomedicine held in Moscow, December, 2007.     

Aggression,Digit Ratio,and Variation in the Androgen Receptor,Serotonin Transporter,and Dopamine D4 Receptor Genes in African Foragers: The Hadza

The role of genes in the expression of aggression and masculinity traits in humans has been a focus of recent behavioral genetic studies. This is the first study on the variation in aggression, the digit ratio (the ratio between the second and the fourth digits, 2D:4D), the directional asymmetry in 2D:4D (D_R-L) and polymorphisms of the AR, DRD4, and 5-HTTL genes in simple hunter-gatherers, namely the Hadza of Tanzania (142 adult men). The distribution of AR, DRD4E3, and 5-HTTLPR genotypes and allele frequencies in Hadza was compared to other African populations on which the data were available. Hadza and Ariaal differed significantly in the distributions of frequencies of AR alleles with different numbers of CAG repeats. Hadza population was similar to other African populations in the distribution of allelic frequencies of the DRD4E3 locus, and to Afro-Americans in the distribution of allelic types of the 5-HTTLPR locus. We found no influence of AR gene on the right hand 2D:4D ratio, D_R-L, and any of aggression subscales of the Buss-Perry Aggression Questionnaire (AQ). Although, a weak positive correlation between CAG repeats and the left hand 2D:4D was found. The multiple regression analysis with digit ratios, D_R-L and aggression subscales of AQ as dependent variables and the three gene candidates (AR, DRD4E3, and 5-HTTLPR) as independent variables revealed the following: men with lower number of CAG repeats had significantly lower left hand 2D:4D ratio; men with higher numbers of 48-bp unit copies in exon 3 of a VNTR polymorphism in the DRD4 gene had significantly lower digit ratios on both hands; no effect of the 5-HTTLPR gene on either the digit ratio or aggressive behavior. These findings demonstrate the complexity of gene effects on digit ratios and aggression and call for simultaneous analysis of more candidate genes. It is noteworthy that these results were obtained for a human population that is still practicing foraging and has been subjected to a high selective pressure due to harsh environments and practically has no access to modern medical care. Hadza are highly egalitarian, and their culture does not favor persons with a dominant or aggressive behavior. It is still to be found to what extent the relationships observed in this study are similar to those in other human populations.