Physiology and Morphological Correlates of Excitatory Transmission are Preserved in Glutamine Transporter SN1-Depleted Mouse Frontal Cortex.

Glutamine is an astroglia-derived precursor of the neurotransmitter glutamate, and its astroglia-to-neuron transfer is controlled by distinct glutamine transporters on the astrocytic and neuronal sites. In this study, we focused on the role of astrocytic glutamine efflux-mediating system N transporter SN1 in the maintenance of glutamatergic neurotransmission by analyzing the electrophysiological parameters ex vivo in the brain slices from control mice and mice in which vivo-morpholino technique was used to diminish SN1 protein. The glutamatergic transmission was characterized by electrophysiological recordings, ultrastructure of neuron terminals, and determination of proteins related to glutamate synaptic transmission: synaptophysin, synaptotagmin, and vit1A. The space-restricted ∼51,5% reduction of SN1 protein did not affect the expression of the neuronal glutamine transporter SAT2. SN1 depletion resulted in a reduction of field potentials (FPs), unaltered frequency of spontaneous and miniature excitatory postsynaptic currents (sEPSCs/mEPSCs), and presented a tendency towards a decrease of long-term potentiation (LTP). Ultrastructurally, preserved number of synaptic vesicles, primarily localized centrally of the cell body, correlates with unchanged levels of synaptic proteins. Collectively, the study indicates that glutamatergic transmission proceeds relatively independently of the SN1 - mediated glutamine transfer to the synapse.

[Navigation-assisted sonography for soft tissues in the head and neck region]. / Navigationsunterstützte Weichteilsonographie für den Kopf-Hals-Bereich.

BACKGROUND: In soft tissue surgery of the head and neck region tissue shifts limit the usefulness of conventional CT/MRI-based navigation procedures. Furthermore, changes caused by invasive measures cannot be visualized. METHODS: A novel navigation device for sonography of soft tissues was developed. This consists of a navigated ultrasound scanner, a navigated surgical instrument, and a personal computer with custom-made software. Its use makes an additional visualization by means of CT or MRI dispensable. RESULTS: The system deviation (three-dimensional error) of this newly developed prototype was less than 1 mm. The practical application in a model setup showed good handling properties of the system. Orientation and approach of the surgical instrument to the sonographically visualized target structure were rapid and accurate. CONCLUSION: This new navigation system does not require additional CT or MRI images. The navigated ultrasound probe shows tissue changes in real time. This navigation system is especially suitable for invasive procedures in soft tissues.

Accuracy of experimental mandibular osteotomy using the image-guided sagittal saw.

The aim of this study was to perform an objective assessment of the accuracy of mandibular osteotomy simulations performed using an image-guided sagittal saw. A total of 16 image-guided mandibular osteotomies were performed on four prefabricated anatomical models according to the virtual plan. Postoperative computed tomography (CT) image data were fused with the preoperative CT scan allowing an objective comparison of the results of the osteotomy executed with the virtual plan. For each operation, the following parameters were analyzed and compared independently twice by two observers: resected bone volume, osteotomy trajectory angle, and marginal point positions. The mean target registration error was 0.95±0.19mm. For all osteotomies performed, the mean difference between the planned and actual bone resection volumes was 8.55±5.51%, the mean angular deviation between planned and actual osteotomy trajectory was 8.08±5.50°, and the mean difference between the preoperative and the postoperative marginal point positions was 2.63±1.27mm. In conclusion, despite the initial stages of the research, encouraging results were obtained. The current limitations of the navigated saw are discussed, as well as the improvements in technology that should increase its predictability and efficiency, making it a reliable method for improving the surgical outcomes of maxillofacial operations.

Hepatitis E virus: complete genome sequence and phylogenetic analysis of a Nepali isolate.

The complete nucleotide sequence of the Nepali strain TK15/92 of hepatitis E (HEV) was determined. It showed the highest sequence homology with the Burmese B1 strain, but closer evolutionary relatedness to the Indian strains. Difficulties in reverse-transcribing and amplifying the hypervariable region in ORF1 suggested that strong secondary structures might be intrinsically responsible for the high mutational rate observed in this region of the HEV genome.

Navigation-supported and sonographically-controlled fine-needle puncture in soft tissues of the neck.

In surgery, sonography has been a well-accepted means of orientation for years. The immediate vicinity of many vital structures in the head and neck region calls for a very exact visualization of the surgical instrument in the 2-D ultrasonic picture. We report on the development of a new method for navigation-supported and sonographically-controlled fine-needle puncture in soft tissues of the neck. Our system comprises a navigated ultrasound probe, a navigated fine-puncture needle and a coordinate sensor. A personal computer with specially-developed software assists calibration and surgical application. The applicability test for the system is described. In vitro, a model lymph node of 9 mm in diameter had been hit. It is shown that the target structure can be aimed at very precisely by the navigated puncture needle. An accuracy of 97% and a specificity of 99% could be demonstrated. The development of a very precise and easy-to-handle method for navigation-supported fine-needle puncture in the neck region is presented. The outstanding advantage of this method is that no rigid reference gadget fixed to the patient's body is necessary. That makes this method very suitable for surgery in the neck region. Contrary to other sonographically-supported navigation methods in the head and neck region, preoperative imaging (CT or MRT) is dispensable.

Complete DNA sequence of Yersinia enterocolitica serotype 0:8 low-calcium-response plasmid reveals a new virulence plasmid-associated replicon.

The complete nucleotide sequence and organization of the Yersinia enterocolitica serotype 0:8 low-calcium-response (LCR) plasmid, pYVe8081, were determined. The 67,720-bp plasmid encoded all the genes known to be part of the LCR stimulon except for ylpA. Eight of 13 intact open reading frames of unknown function identified in pYVe8081 had homologues in Yersinia pestis plasmid pCD1 or in Y. enterocolitica serotype 0:9 plasmid pYVe227. A region of approximately 17 kbp showed no DNA identity to pCD1 or pYVe227 and contained six potential new genes, a possible new replicon, and two intact insertion sequence (IS) elements. One intact IS element, ISYen1, was a new IS belonging to the IS256 family. Several vestigial IS elements appeared different from the IS distribution seen in the other LCR plasmids. The RepA proteins encoded by Y. enterocolitica serotype 0:8 pYVeWA and pYVe8081 were identical. The putative pYVe8081 replicon showed significant homology to the IncL/M replicon of pMU407.1 but was only distantly related to the replicons of pCD1 and pYVe227. In contrast, the putative partitioning genes of pYVe8081 showed 97% DNA identity to the spy/sopABC loci of pCD1 and pYVe227. Sequence analysis suggests that Yersinia LCR plasmids are from a common ancestor but that Y. enterocolitica serotype 0:8 plasmid replicons may have evolved independently via cointegrate formation following a transposition event. The change in replicon structure is predicted to change the incompatibility properties of Y. enterocolitica serotype 0:8 plasmids from those of Y. enterocolitica serotype 0:9 and Y. pestis LCR plasmids.

Sequencing of the Francisella tularensis strain Schu 4 genome reveals the shikimate and purine metabolic pathways, targets for the construction of a rationally attenuated auxotrophic vaccine.

Francisella tularensis is the etiological agent of tularemia, a serious disease in several Northern hemisphere countries. The organism has fastidious growth requirements and is very poorly understood at the genetic and molecular levels. Given the lack of data on this organism, we undertook the sample sequencing of its genome. A random library of DNA fragments from a highly virulent strain (Schu 4) of F. tularensis was constructed and the nucleotide sequences of 13,904 cloned fragments were determined and assembled into 353 contigs. A total of 1.83 Mb of nucleotide sequence was obtained that had a G+C content of 33.2%. Genes located on plasmids pOM1 and pNFL10, which had been previously isolated from low virulence strains of F. tularensis, were absent but all of the other known F. tularensis genes were represented in the assembled data. F. tularensis Schu4 was able to grow in the absence of aromatic amino acids and orthologues of genes which could encode enzymes in the shikimate pathway in other bacteria were identified in the assembled data. Genes that could encode all of the enzymes in the purine biosynthetic and most of the en- zymes in the purine salvage pathways were also identified. This data will be used to develop defined rationally attenuated mutants of F. tularensis, which could be used as replacements for the existing genetically undefined live vaccine strain.