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Differential expression of the PSD-95 gene family in electrosensory neurons.

The PSD-95 family of membrane-associated guanylate kinase (MAGUK) proteins are involved in the assembly and organization of neurotransmitter receptors at excitatory synapses in the vertebrate nervous system. We have isolated partial cDNAs for five PSD-95 family members from Apteronotus leptorhynchus brain RNA using a degenerate PCR method. The amino acid sequences deduced indicate that A. leptorhynchus neurons express homologues of the mammalian PSD-93, SAP-97, and SAP-102 MAGUKs and two homologues of mammalian PSD-95. In situ hybridization experiments have been carried out to localize the cellular expression of all five MAGUK mRNAs in the central nervous system of A. leptorhynchus. In the cerebellum the expression patterns are highly similar to patterns reported for mammalian cerebellum, suggesting an evolutionary conservation of the functional roles in this gene family. Cellular levels of expression of the PSD-95 MAGUK mRNAs and the NMDAR-1 mRNA were highly correlated in neurons of the dorsal forebrain but were not correlated in neurons of the electrosensory lateral line lobe (ELL) or the cerebellum. These results suggest that the expression of PSD-95 MAGUK genes in forebrain neurons may provide mechanisms for synaptic organization that are not shared by neurons in the ELL and cerebellum.

Amino Acid Sequence↗

On-line aSi portal imaging of implanted fiducial markers for the reduction of interfraction error during conformal radiotherapy of prostate carcinoma.

PURPOSE: An on-line system to ensure accuracy of daily setup and therapy of the prostate has been implemented with no equipment modification required. We report results and accuracy of patient setup using this system. METHODS AND MATERIALS: Radiopaque fiducial markers were implanted into the prostate before radiation therapy. Lateral digitally reconstructed radiographs (DRRs) were obtained from planning CT data. Before each treatment fraction, a lateral amorphous silicon (aSi) portal image was acquired and the position of the fiducial markers was compared to the DRRs using chamfer matching. Couch translation only was used to account for marker position displacements, followed by a second lateral portal image to verify isocenter position. Residual displacement data for the aSi and previous portal film systems were compared. RESULTS: This analysis includes a total of 239 portal images during treatment in 17 patients. Initial prostate center of mass (COM) displacements in the superior, inferior, anterior, and posterior directions were a maximum of 7 mm, 9 mm, 10 mm and 11 mm respectively. After identification and correction, prostate COM displacements were <3 mm in all directions. The therapists found it simple to match markers 88% of the time using this system. Treatment delivery times were in the order of 9 min for patients requiring isocenter adjustment and 6 min for those who did not. CONCLUSIONS: This system is technically possible to implement and use as part of an on-line correction protocol and does not require a longer than standard daily appointment time at our center with the current action limit of 3 mm. The system is commercially available and is more efficient and user-friendly than portal film analysis. It provides the opportunity to identify and accommodate interfraction organ motion and may also permit the use of smaller margins during conformal prostate radiotherapy. Further integration of the system such as remote table control would improve efficiency.

Humans↗

Supraspinal cerebral areas involved in morphine's intestinal inhibition and analgesia.

To explore the neuroanatomical pathways involved in mediating the antipropulsive effect and analgesia of morphine (M) in the periaqueductal gray matter (PAG), we examined the influence of the vagus nerve and the role of serotonergic neurotransmission. M-induced inhibition of intestinal transit was unaffected by subdiaphragmatic vagotomy. In contrast, electrolytic lesions in the raphe magnus nucleus (NRM) and pretreatment with a selective neurotoxin (5,6-DHT, 15 micrograms/rat) in the same region, both significantly reduced M-induced inhibition of intestinal transit. Analgesia was only slightly affected. p-CPA pretreatment (100 mg/kg IP) induced the same results. Finally some other central brain regions were found to be sensitive to M's intestinal inhibition and analgesia such as the mid-line thalamus, the dorsal and lateral hypothalamus, and the bulbar reticular formation. Negative results were obtained for frontal cortex, caudate and amygdala. Some considerations are put forward about the existence in the central nervous system of selective areas involved in intestinal modulation and their relation with those mediating pain transmission.

Analgesia↗

The xipotl mutant of Arabidopsis reveals a critical role for phospholipid metabolism in root system development and epidermal cell integrity.

Phosphocholine (PCho) is an essential metabolite for plant development because it is the precursor for the biosynthesis of phosphatidylcholine, which is the major lipid component in plant cell membranes. The main step in PCho biosynthesis in Arabidopsis thaliana is the triple, sequential N-methylation of phosphoethanolamine, catalyzed by S-adenosyl-l-methionine:phosphoethanolamine N-methyltransferase (PEAMT). In screenings performed to isolate Arabidopsis mutants with altered root system architecture, a T-DNA mutagenized line showing remarkable alterations in root development was isolated. At the seedling stage, the mutant phenotype is characterized by a short primary root, a high number of lateral roots, and short epidermal cells with aberrant morphology. Genetic and biochemical characterization of this mutant showed that the T-DNA was inserted at the At3g18000 locus (XIPOTL1), which encodes PEAMT (XIPOTL1). Further analyses revealed that inhibition of PCho biosynthesis in xpl1 mutants not only alters several root developmental traits but also induces cell death in root epidermal cells. Epidermal cell death could be reversed by phosphatidic acid treatment. Taken together, our results suggest that molecules produced downstream of the PCho biosynthesis pathway play key roles in root development and act as signals for cell integrity.

Arabidopsis↗

Comparative anatomy of cerebellar catecholamine innervation from teleosts to mammals.

The cerebellar innervation of the catecholamine (CA) neurons was experimentally investigated with histofluorescence method in the brains of the vertebrates from teleosts to mammals. In the teleosts the source for the cerebellar CA are composed of three CA neuron groups. Hypothalamic CA neuron group contributes mainly to innervation in the valvula cerebelli whose development is dependent on the high degree of the development of the lateral-line organ. CA neurons in the medulla oblongata might innervate the vestibulo-lateral lobe and corpus cerebelli. These two CA neuron groups appear to constitute the main source for the cerebellar CA, though it is certain that rhombencephalic CA neurons also send their axons to innervate the cerebellum. In the tailless amphibian which is devoid of the lateral-line nerves, cerebellar CA especially in the corpus cerebelli is supplied mainly by the rhombencephalic CA neurons which still remain in less developed stage in phylogenetic scale. In the bird, ascending CA neuron system is maintained by long axons and the anlage of locus coeruleus (LC) appear first in this class. A large number of CA nerve terminals in the cerebellum of the bird are supplied by both CA neurons of the dorsal part of the tegmentum and CA neurons disseminated in the tegmental reticular formation. In the mammals showing the highest development of the cerebellar and cerebral cortices, LC is well developed and cerebellar CA is supplied mainly by LC, though cerebellum is innervated by other CA neurons besides LC. Possible significance of the CA neurons on the development of the cerebellum in phylogeny was discussed.

Animals↗

[Changes in the subendothelial compartment during the maturation process of cerebral microvessels].

Basement lamina and pericytes of growing blood microvessels were analyzed in the chick embryo optic tectum, from the 8th incubation day to hatching. Formation of the basement lamina and morphological changes of the pericytes take place in a short range of time, but late in the embryonic life, when also the blood brain barrier (bbb) devices are developing. The spatial and temporal coincidence between basement lamina formation, endothelium tight junction differentiation, and perivascular arrangement of the astrocytic glia, indicates that these events are correlated and corroborates the hypothesis that the glia needs an extracellular matrix to induce the junctional system maturation in the neural endothelia. Pericytes are irregular in shape during the early neural angiogenesis and smooth and flattened later, as the basement lamina synthesis is taking place; these cells represent a second line of barrier beyond the endothelium when the bbb is immature, owing to their phagocytic and digestive properties.

Animals↗

Autopsy case of thanatophoric dysplasia: observations on the serial sections of the brain.

The neuropathological findings in an autopsy case of thanatophoric dysplasia (TD) with serial sections of the brain are described here. This patient was a female infant, born at 33 weeks gestation, who died on day 1. Skeletal anomalies, consisting of short limbs, a small thorax, short ribs, thick cortical vertebral body substance and sternum substance, and hypoplastic lungs, were compatible with typical phenotypic features of TD. The brain weighed 370 g, showing a cloverleaf megalencephaly. A computerized 3-D reconstruction technique visualized clearly abnormal deep sulci arranged perpendicular to the neuraxis on the inferior surface of the temporal lobe, and peculiar configurational changes of the lateral ventricle. In particular, the inferior horn showed an unusual complex form. Dysgenetic changes were largely located in the anterior temporal lobe as follows: cortical polymicrogyria; leptomeningeal heterotopia with discontinuity of the subpial basement membrane; serpentine arrangement of pyramidal cells of the cornu ammonis (CA)1 of the hippocampus; hypoplastic dentate gyrus; hyperplasia of the amygdaloid body; and heterotopic nodules of neuroblasts or glioblasts in the periventricular white matter. Apart from the temporal lobe, the cerebral pia mater showed unusual fusion of two facing sheets in a sulcus and ectopia of nerve cells, and the cerebellar vermis was small. The findings observed here indicate that overgrowth and lack of growth can coexist in the TD brain, suggesting that some interaction(s) between the mesenchyme and the nervous tissue may play a role in normal differentiation of these two cell lines.

Bone and Bones↗

Simulation of ST segment changes during subendocardial ischemia using a realistic 3-D cardiac geometry.

The mechanisms underlying the ST segment shifts associated with subendocardial ischemia remain unclear. The aim of this paper is to shed further light on the subject through numerical simulations of these shifts. A realistic three-dimensional model of the ventricles, including fiber rotation and anisotropy, is embedded in a nonhomogeneous torso model. A simplification of the bidomain model is used to calculate only the ST segment shift, assuming known values of the transmembrane potential during the plateau and rest phases. A similar simulation is performed in two dimensions. The simulation results suggest that subendocardial ischemia can be located by ST segment shift on the epicardial and torso surfaces. It is shown that ST elevation is associated with the transmural ischemic boundary, while ST depression is associated with the lateral ischemic boundaries.

Body Surface Potential Mapping↗

[Implementation of transabdominal ultrasound image guided conformal intensity modulated radiotherapy].

PURPOSE: Optically guided ultrasound imaging has been used in our department since 2003 in order to implement an on line correction scheme in intensity modulated radiation therapy of prostate carcinoma. PATIENTS AND METHODS: The corrections observed during the initial time period of the system (17 patients) are compared to those observed more recently (10 patients). Treatment margins are calculated. RESULTS: Overall systematic errors decreased between 2003 and 2006, and are presently statistically not different from zero. Random errors remain the same (max 4.3 mm). Proposed margins are 7 mm both in lateral and longitudinal direction and 8.4 mm in anteroposterior. CONCLUSION: Ultrasound can be used for on line correction of both positioning and internal organs motion errors and allows reduction of the margins between clinical and planning volume.

Abdomen↗

Specific order in the appearance of protein tyrosine phosphorylation patterns is functionally coordinated with dog sperm hyperactivation and capacitation.

The aims of the present study were to characterize a slow capacitation system that records initial changes in the sperm membrane state, and, using a canine model, to order the specific protein tyrosine phosphorylation signaling in the sequence of capacitational events and to associate them with hyperactivated motility. Dog sperm washed through Percoll were incubated in complete bicarbonate Tyrode medium for 6 hours in 5% CO(2). Capacitation was evaluated using chlortetracycline staining. Tyrosine phosphorylation patterns were assessed by immunocytochemistry. Parallel to this, a computer-assisted motility analysis was performed. Significant changes in the percentage of capacitated and acrosome-reacted cells were first observed after 90 minutes, increasing in a linear manner during further incubation (P <.05). Changes in the percentage of capacitated cells were accompanied by motility changes. During incubation, a strictly sequential phosphorylation of sperm tail (midpiece, principal piece, and end piece) and head proteins was observed. According to an analysis of kinetics, phosphorylation of head proteins occurred after the tail became completely phosphorylated. Changes in head phosphorylation progressed at the same rates as capacitation and acrosome reaction. Sperm motility, curvilinear velocity, average path velocity, straight line velocity, and lateral head displacement were correlated positively or negatively with phosphorylation of midpiece or end piece proteins, respectively. The bicarbonate-stimulated increases in cyclic adenosine monophosphate levels and changes in protein phosphatase activity may be involved in the signaling system that controls membrane changes and motility in dog sperm. Phosphorylation kinetics of sperm proteins are potentially useful for diagnostic purposes to characterize the response of individual males to fertilizing conditions.

Animals↗

[Preoperative autologous blood donation, effect of hydroxyethyl starch on the reticuloendothelial system and opsonins].

The effect of 6% low molecular weight (LMW) HES (MW 270,000) on reticuloendothelial function was studied in 9 male patients, median of age 50 years, who were scheduled for vascular surgery. An erythrocyte clearance test (Anti Rh) labeled with 99 mTc, evaluating primarily the splenic section of the RES, showed a considerable variation (37-87%) prior to hemodilution. 24 hours after replacement of 15 ml/kg body weight of blood, there was no significant change of the clearance rate, a follow up weeks later approached the base line values. Plasma opsonins like fibronectin, complement fraction 3, complement fraction 4 and immunoglobulin G showed a proportional decrease following hemodilution, and after 24 hours approached control levels. Changes at this time seemed to be independent from the course of the hematocrit. It is concluded that LMW HES does not adversely influence the spleen dependent phagocytic capacity of the RES system.

Blood Transfusion, Autologous↗

[The minimally invasive stabilization of the dorsal pelvic ring with the transiliacal internal fixator (TIFI)--surgical technique and first clinical findings].

The Trans Iliacal Internal Fixator (TIFI) is a minimally invasive technique for the stabilization of sacro-iliac joint ruptures and fractures lateral to the sacral ala or through the sacral foramen. In this study, 7.0 mm pedicle screws of the Universal-Spine-System (USS, Synthes) were inserted 1-2 cm on the cranial side of the posterior superior iliac spine and parallel to the superior gluteal line. The connecting bar was inserted subfascially and fixed with the locking head pedicle screws to form an fixed-angle construction. In a prospective study 31 patients with vertical shear injuries of the pelvis were treated with the TIFI. There were two wound infections and one loosening of a pedicle screw. None of the screws were incorrectly positioned and no neuro-vascular lesions were caused by the implant. 2 years postoperatively we found 50% good and excellent results for type C pelvic ring injuries. Early findings show that the TIFI is well suited to stabilization of sacro-iliac joint ruptures and fractures of the lateral sacrum. Closed reduction and minimally invasive insertion technique are possible. The implant leads to sufficient biomechanical stability but there is a very low intraoperative risk of neuro-vascular lesion.

Adolescent↗

Differential alcohol drinking behaviour and dependence in the Naples low- and high-excitability rat lines.

Alcoholism is composed of a series of phenomena, which can be approached in model systems. Our aim was to investigate psychogenetics of alcohol drinking behaviour and dependence in the Naples rat lines. Thus, adult male rats of the Naples low- (NLE), high-excitability (NHE) and random-bred control (NRB) lines had 24-h access to a four-bottle system with tap water and 5, 10 and 20% alcohol solutions. Total voluntary alcohol intake and preference were measured during a 6-week period. In order to analyze behavioural dependence, two abstinence periods were introduced, followed by re-exposure to the bottles. The first was a pure re-test and the second was a quinine test (adding 0.02% quinine alcohol solution). Finally, to verify the role of opioids on alcohol drinking behaviour, rats received naltrexone (5 mg/kg i.p.). Thirty minutes later, they had access for 2 h to a three-bottle system with water, 5 and 20% alcohol solution, respectively. Results indicate that (i) total alcohol intake on day 1 was higher in both NLE/NHE, becoming higher in NLE rats during the 6-week period, (ii) different alcohol preference patterns in the Naples lines were observed over time of testing, (iii) during re-test, NLE rats showed neither reversibility nor alcohol deprivation effect, (iv) during quinine test, only NLE rats reduced alcohol intake to a lower extent and (v) naltrexone reduced by 50% the intake of the preferred solution. Therefore, the NLE rats appear as a new model for studying the neural substrates of the dependence behaviour.

Alcohol Drinking↗

Global right atrial mapping delineates double posterior lines of block in patients with typical atrial flutter: a study using a three dimensional noncontact mapping system.

UNLABELLED: Double Posterior Lines of Block in Typical Atrial Flutter. INTRODUCTION: The crista terminalis (CT) has been shown to be a barrier to transverse conduction during typical atrial flutter (AFL). However, some studies have demonstrated the presence of functional block in the sinus venosa region but not at the CT. The aim of this study was to define these regions of block in the right atrium using a three-dimensional noncontact mapping system. METHODS AND RESULTS: In 39 AFL patients (33 men and six women, mean age 56 +/- 13 years), a noncontact multielectrode array was used to reconstruct electrograms in the right atrium. Isochronal and isopotential propagation mapping was performed during AFL and during pacing from the coronary sinus ostium and the low lateral wall (cycle length from 600 to 240 msec) in sinus rhythm after creation of isthmus block. A single line of block along the CT area was found in 18 patients (46%). Two lines of block were found in 21 patients (54%), with the first line located along the CT area. The second was located in the sinus venosa region in 20 patients (51%) and in the lateral wall in 1 patient (3%). In all patients, the block in the lower part of the CT was observed during AFL (60%) and during pacing at all cycle lengths (48%-62%). The length and proportion of block were inversely proportional to pacing cycle length. CONCLUSION: Double lines of block were frequently observed in patients with AFL, and both lines may form the posterior boundaries of the AFL circuit. Block was fixed in the lower part of the CT and was functional in the upper part of the CT.

Atrial Flutter↗

Emergence of resistance to VP-16/cisplatin chemotherapy: study in a lymphoblast model system.

Etoposide (VP-16) and cisplatin are widely used in the treatment of malignancy. It is a common clinical observation that patients may initially respond to this two drug combination but later become resistant to it. Data from the CCL-159 lymphoblast cell line suggests that the emergence of resistance may be by means other than multiple drug resistance gene expression. The data suggest that chemotherapeutic failure may be mediated in part by a relatively inefficient method of drug resistance, the unstable expression of low-level drug resistance by a minority of cells. These results may help explain how patients whose malignancies are initially sensitive to VP-16/cisplatin later develop drug resistance.

Antineoplastic Combined Chemotherapy Protocols↗

Three-dimensional line-field Fourier domain optical coherence tomography for in vivo dermatological investigation.

We demonstrate 3-D optical coherence tomography using only 1-D mechanical scanning. This system uses the principle of Fourier domain optical coherence tomography for depth resolution, 1-D imaging for lateral vertical resolution, and mechanical scanning by a galvanometer for lateral horizontal resolution. An in vivo human fingerpad is investigated in three dimensions with an image size of 480 points (vertical) x 300 points (horizontal) x 1024 points (depth), which corresponds to 2.1 x 1.4 x 1.3 mm. The acquisition time for a single cross section is 1 ms and that for a single volume is 10 s. The system sensitivity is 75.6 dB at a probe beam power of 1.1 mW.

Dermoscopy↗

Generalization of induced interhemispheric interference: a chronometric approach to one-brain vs two-brains models of the hemispheres' cooperation.

A reaction time study with normal human subjects was conducted to obtain support for one or the other model of interhemispheric relations--referred to as the "one-system" and the "two-systems" hypotheses. 12 subjects were extensively trained in a complex reaction time task consisting of a "priming sub-task" which introduced interhemispheric interference and two different "test sub-tasks" measuring the generalization of interference. The interference priming produced visible slowing of RT's on subsequent trials. Interestingly, the deterioration of the two test sub-tasks was alike, despite the marked difference in the amount of the interhemispheric communication they required. This result is more in line with the one-system hypothesis, as the two-systems hypothesis predicts deterioration proportional to the amount of the required interhemispheric communication.

Adult↗

Lateralized regular spatial patterns in oscillating drawing arm movements of right-handed young women.

There is a lacuna in literature with reference to the spatial lateral difference in fast rhythmical movements produced by the whole dominant and nondominant whole arm, where spinal regulation has a significant role. Based on a fast oscillating zigzag drawing task, this study focused on (a) creation of a specific model of the task based on the intermittencies of coupled vectors of the fast motion, (b) identification of the spatial patterns that triggered these vectors, and (c) identification of quantified lateral differences between the spatial rhythmical patterns. 12 strongly right-handed young women performed 9 to 11 trials drawing zigzag lines. Each participant was required to extend her arm and perform this task using the left and right arm selectively on a frontally positioned graphic design system. The spatial patterns produced on each trial were identified in terms of five constant combinations of horizontal (X) and vertical (Y) projections of each line on the zigzag drawings. The dominant arm differed from the nondominant arm in preferred patterns. Because the duration of each line in the zigzag was highly restricted in time, the appearance of the patterns with different block schemes of movement could be explained as being associated with lower levels of the central nervous system. Initiation of fast movement of the total upper arm is probably associated with selection of the block scheme of motor control appropriate to each arm. Each block scheme is grounded on the coupled vectors of motion organised with particular muscle groups. Some block schemes seemed linked specifically to the dominant arm.

Adult↗