Gene silencing in rat-liver and limb grafts by rapid injection of small interference RNA

Genetic modification is a promising therapeutic strategy for organ transplantation. In previous studies, we introduced a nonviral method of gene transfer to donor grafts using an organ-selective injection technique to up-regulate gene expression. Based on the attractive methodology of RNA interference for silencing a particular gene expression, we applied our catheter-based injection method to transfer small interference RNA (siRNA)-green fluorescence protein (GFP) into liver and limb grafts. We first quantified the interfering activity after the systemic delivery of siRNA in the liver of Alb-DsRed2 transgenic (Tg) rats using in vivo bioimaging system. Then, using GFP Tg Lewis rats as donors, transient down-regulation of the GFP expression was achieved both in liver- and limb-transplantation models by the preoperative rapid injection of siRNA. Genetic modification by siRNA may provide new therapeutic options for down-regulation of endogenous antigenicity.     

Image-guided surgery for thoracic ossification of the posterior longitudinal ligament. Technical note

The authors describe an anterior decompression procedure for thoracic ossification of the posterior longitudinal ligament (PLL) in which they used an image guidance system in three cases. To make registration possible in anterior thoracic surgery, they devised a surgical reference frame that could be connected to a rod and attached to an external fixation device, which was then attached to the thoracic VB. The mean fiducial error at the registration was acceptable (range 0.5-0.8 mm). They were able to confirm the success of decompression on postoperative computerized tomography scans. In the removal of an ossified thoracic PLL, an image guidance system has been shown to be a useful tool.     

Diffuse large B-cell lymphoma with extra Bcl-2 gene signals detected by FISH analysis is associated with a "non-germinal center phenotype"

Amplification and translocation of the Bcl-2 gene has been detected in a certain subset of diffuse large B-cell lymphomas (DLBCL). The correlations among Bcl-2 protein expression, gene translocation or amplification, and the molecular signature determined by cDNA array are poorly understood. This study examined 25 cases with de novo nodal DLBCL. Interphase fluorescence in situ hybridization (FISH) analysis was performed to evaluate the Bcl-2 gene using IGH/BCL2 and CEP18 centromere probes (Vysis). When extra Bcl-2 gene signals were observed in each tumor cell and when these signals were in proportion to the extra CEP18 probe signals, we regarded the findings as indicating the presence of an additional chromosome 18; when extra Bcl-2 signals were observed but additional CEP18 signals were not, we regarded the findings as indicating the presence of gene amplification. A panel of 3 antigens (CD10, Bcl-6, and MUM-1) was applied to categorize each case as either a "germinal center B-cell (GCB) phenotype" or a "non-GCB phenotype." Of the 25 cases examined, 8 cases (32%) were classified as "GCB phenotype" and 17 cases (68%) were classified as "non-GCB phenotype." A FISH analysis revealed that t(14;18) was detected in 2 of the 8 cases (25%) with the "GCB phenotype" but in none of the 17 "non-GCB phenotype" cases. Extra Bcl-2 gene signals were detected in 7 of the 25 (28%) cases examined: n = 5 for an additional chromosome 18, n = 1 for gene amplification, and n = 1 for additional chromosome 18 + gene amplification. Extra Bcl-2 gene signals were exclusively detected in DLBCL with the "non-GCB phenotype"; these cases, with the exception of one, stained strongly positive for Bcl-2. The DLBCLs with Bcl-2 protein overexpression were classified into at least two heterogeneous molecular groups, based on the results of the FISH analysis.     

Investigation about the homogeneity of nasopharyngeal microflora at the different location of nasopharynx of children with acute otitis media

OBJECTIVE: Nasopharynx is thought to be a very important site as becterial reservoir for acute otitis media (AOM). In this study, we investigated on the homogeneity of nasopharyngeal microflora at the different location of nasopharynx of children with AOM. METHODS: Thirty nasopharyngeal samples of 15 children with AOM, two samples harvested from both nostrils of each child, were cultured and analyzed by patterns of antibiotic susceptibility and pulsed-field gel electrophoresis analysis (PFGE). RESULTS: A total of 30 nasopharyngeal samples were cultured and 19 isolates of Streptococcus pneumoniae from 10 children (66.7%), 8 isolates of Haemophilus influenzae from 4 children (26.7%), and 12 isolates of Moraxella catarrhalis from 7 children (46.7%) were obtained. In all children except three, the nasopharyngeal microflora at right and left orifice of the eustachian tubes showed no obvious differences in the bacterial species and quantities. Furthermore, in children with the same species of were cultured from right and left orifice of the eustachian tubes at the same time, all nine couples of S. pneumoniae isolates, four couples of H. influenzae isolates, and five couples of M. catarrhalis isolates showed about the same susceptibility and PFGE patterns. CONCLUSION: These results suggest that the microflora at the different location of nasopharynx of children with AOM is almost homogeneous, irrespective of the clinical signs.     

Transabdominal measurement of oxygenation of the placenta by near-infrared spectroscopy

Near-infrared spectroscopy (NIRS) has been used as a noninvasive method for monitoring the real-time oxygenation status in areas such as the brain and striated muscle. Because the oxygenation status of the placenta is closely related to the fetal condition, monitoring placental oxygenation through the maternal abdomen is desirable. We performed transabdominal monitoring of oxygenation of the placenta by NIRS in 11 women. We improved the conventional probe of the NIRO 300 system (Hamamatsu Photonics KK, Hamamatsu, Japan) to obtain placental oxygenation data. With this probe, we succeeded in obtaining oxyhemoglobin and deoxyhemoglobin data through the maternal abdomen. We believe NIRS will prove to be useful for intrapartum monitoring.