Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa

The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice generated by crossing CaMKIIα-Cre+/Cre mice with floxed ROSA26 lacZ reporter (Rosa26+/stop-lacZ) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice. Similarly, when double transgenic Gnao+/f::CaMKIIα-Cre+/Cre mice carrying a floxed Go-alpha gene (Gnaof/f) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (GnaoΔ) without inheriting the Cre transgene. The GnaoΔ/Δ mice closely resembled conventional Go-alpha knockout mice (Gnao−/−) with respect to impairment of their behavior. Thus, we conclude that CaMKIIα-Cre mice afford recombination for both tissue- and time-controlled inactivation of floxed target genes in the brain and for their permanent disruption. This work also emphasizes that extra caution should be exercised in utilizing CaMKIIα-Cre mice as breeding pairs.Fil: Choi, Chan-Il. Ajou University. School of Medicine; Corea del SurFil: Yoon, Sang-Phil. Ajou University. School of Medicine; Corea del SurFil: Choi, Jung-Mi. Ajou University. School of Medicine; Corea del SurFil: Kim, Sung-Soo. Ajou University. School of Medicine; Corea del SurFil: Lee, Young-Don. Ajou University. School of Medicine; Corea del SurFil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Suh-Kim. Haeyoung. Ajou University. School of Medicine; Corea del Su

Allele Frequency and Genotype Distribution of STR SE33 locus in Koreans

DNA was obtained from placental tissue and peripheral blood of 268 unrelated Koreans in order to analyze the applicability of PCR amplification of human beta-actin related pseudogene H-beta-Ac-psi-2 (AClBP-2, SE33) SlR locus for forensic medicine in the field of individual identification and paternity tests. After amplification using two primers specific to the SE33 locus, the products were analyzed by polyacrylmide gel electrophoresis, so called Amp-FLP(Amplification Fragment Length Polymorphism) procedure followed by silver staining. To improw the differentiation of the allele an automatic sequencer, Licor 4000-L was used and the result was compared with that of routine acrylamide gel electrophoresis. 36 alleles were deteced, and the distribution was somewhat homogenous, and none of the alleles were above 0.1 in frequency. Allele 12 and allele 14 were most frequent with frequencies of 0.09701 and 0.08022, and alleles of two base repeat such as allele 17, allele 19 were also encountered. 144 kinds of genotypes were found, among which 22 were homozygotes, and the heterozygosity was 91.85%. In 164 gametes of 64 families whose parent-child relationship were confirmed through other studies and information, Mendelian inheritance was well presented although two mutations were observed

Uncertainty of stress path in fault stability assessment during CO<inf>2</inf> injection: Comparing smeaheia 3D geomechanics model with analytical approaches

publishedVersio

Ru-Catalyzed, cis-Selective Living Ring-Opening Metathesis Polymerization of Various Monomers, Including a Dendronized Macromonomer, and Implications to Enhanced Shear Stability

An unsaturated polymer’s cis/trans-olefin content has a significant influence on its properties. For polymers obtained by ring-opening metathesis polymerization (ROMP), the cis/trans-olefin content can be tuned by using specific catalysts. However, cis-selective ROMP has suffered from narrow monomer scope and lack of control over the polymerization (giving polymers with broad molecular weight distributions and prohibiting the synthesis of block copolymers). Herein, we report the versatile cis-selective controlled living ROMP of various endo-tricyclo[4.2.2.0^(2,5)]deca-3,9-diene and various norbornene derivatives using a fast-initiating dithiolate-chelated Ru catalyst. Polymers with cis-olefin content as high as 99% could be obtained with high molecular weight (up to M_n of 105.1 kDa) and narrow dispersity (<1.4). The living nature of the polymerization was also exploited to prepare block copolymers with high cis-olefin content for the first time. Furthermore, owing to the successful control over the stereochemistry and narrow dispersity, we could compare cis- and trans-rich polynorbornene and found the former to have enhanced resistance to shear degradation

Intraocular Involvement of a Nasal Natural Killer T-Cell Lymphoma: A Case Report

Herein, we report a case of nasal natural killer T-cell lymphoma (NKTL) with intraocular involvement. A 57-year-old woman was referred due to a three-day history of photophobia and diplopia in the left eye. One-month previously, she was diagnosed with nasal NKTL of the right nasal cavity. Ophthalmic examination revealed conjunctival injection and ptosis. The left pupil was fully dilated and non-reactive to light. Ocular motion was restricted on left-upper gaze. Five days later, anterior uveitis developed and persisted despite topical steroid treatment. An orbital magnetic resonance imaging was without specific findings, however, ophthalmoplegia, vitreous opacity, and an iris mass were observed. A diagnostic anterior chamber aspiration was performed. Aqueous humor aspiration revealed 35% morphologically atypical lymphocytes. After an intravitreal triamcinolone injection, radiotherapy and chemotherapy were administered; this resolved the uveitis and iris mass. When refractory uveitis or orbital pseudotumor occurs in patients with nasal NKTL, ocular and orbital involvement of the NKTL should be considered

Potential impacts of gas hydrate exploitation on slope stability in the Danube deep-sea fan, Black Sea

Highlights • The Danube deep-sea fan offers best conditions for hydrate production. • Gas production out of a hypothetical methane hydrate reservoir was simulated. • Hazard assessment to investigate the hazard of production-induced slope failures. • Factor of Safety against slope failure is not affected by the production process. • Mobilized mass could hit the production site if landslide were to happen. Methane production from gas hydrate reservoirs is only economically viable for hydrate reservoirs in permeable sediments. The most suitable known prospect in European waters is the paleo Danube deep-sea fan in the Bulgarian exclusive economic zone in the Black Sea where a gas hydrate reservoir is found 60 m below the seafloor in water depths of about 1500 m. To investigate the hazards associated with gas production-induced slope failures we carried out a slope stability analysis for this area. Screening of the area based on multibeam bathymetry data shows that the area is overall stable with some critical slopes at the inner levees of the paleo channels. Hydrate production using the depressurization method will increase the effective stresses in the reservoir beyond pre-consolidation stress, which results in sediment compaction and seafloor subsidence. The modeling results show that subsidence would locally be in the order of up to 0.4 m, but it remains confined to the immediate vicinity above the production site. Our simulations show that the Factor of Safety against slope failure (1.27) is not affected by the production process, and it is more likely that a landslide is triggered by an earthquake than by production itself. If a landslide were to happen, the mobilized sediments on the most likely failure plane could generate a landslide that would hit the production site with velocities of up to 10 m s-1. This case study shows that even in the case of production from very shallow gas hydrate reservoirs the threat of naturally occurring slope failures may be greater than that of hydrate production itself and has to be considered carefully in hazard assessments

MRI Findings of Pericardial Fat Necrosis: Case Report

Pericardial fat necrosis is an infrequent cause of acute chest pain and this can mimic acute myocardial infarction and acute pericarditis. We describe here a patient with the magnetic resonance imaging (MRI) findings of pericardial fat necrosis and this was correlated with the computed tomography (CT) findings. The MRI findings may be helpful for distinguishing pericardial fat necrosis from other causes of acute chest pain and from the fat-containing tumors in the cardiophrenic space of the anterior mediastinum

Ultra sub-wavelength surface plasmon confinement using air-gap, sub-wavelength ring resonator arrays

Arrays of sub-wavelength, sub-10 nm air-gap plasmonic ring resonators are fabricated using nanoimprinting. In near infra-red (NIR) range, the resonator supports a single dipole mode which is excited and identified via simple normal illumination and explored through transmission measurements. By controlling both lateral and vertical confinement via a metal edge, the mode volume is successfully reduced down to 1.3 × 10(−5) λ(0)(3). The advantage of such mode confinement is demonstrated by applying the resonators biosensing. Using bovine serum albumin (BSA) molecules, a dramatic enhancement of surface sensitivity up to 69 nm/nm is achieved as the modal height approaches the thickness of the adsorbed molecule layers