Desensitization of GABA-induced currents in cultured rat hippocampal neurons.

The basic characteristics of desensitization of the GABAA receptor were investigated in cultured rat hippocampal neurons (three days to four weeks in vitro) using whole cell patch clamp techniques. GABA at 10-500 microM was perfused on to neurons for 30 or 60 s, with 60 s intervals of wash with control bath solution between perfusions. Desensitization, evaluated by peak-to-plateau ratio and time constants of current decay (tau), was dose-dependent and culture age-dependent. Desensitization was observed as early as three days in culture, the earliest time tested. At all ages, higher concentrations of GABA induced both larger and faster desensitization. Desensitization was markedly voltage-dependent and decreased with depolarization; peak-to-plateau ratio went from 6.3 to 1.4 and tau went from 4.6 to 26.8 s when holding potentials were changed from -80 mV to +30 mV. Low concentrations of GABA (1-2 microM) perfused for 2-60 s, which did not induce any current, had no effect on the maximal response nor desensitization produced by a subsequent application of 100 microM GABA. This finding suggests that GABA receptors were not desensitized without first being activated.     

Methods for derivation of human embryonic stem cells

The expanded blastocysts, developed from 2PN-stage embryos, are generally divided into three categories: a good blastocyst containing a large and distinguishable inner cell mass (ICM), a blastocyst with a small and distinct ICM, and a blastocyst with a poorly defined ICM. In this study, we introduce methods for the derivation of human embryonic stem cells (hESCs) depending on the quality of the blastocysts. An immunosurgical method was used for the good expanded blastocysts. This method, however, raises the probability of ICM loss in cases of hESC derivation from blastocysts with smaller or indistinct ICMs. Furthermore, this method is also associated with a risk of the contamination of the hESCs with animal pathogens. To overcome these shortcomings, the partial- or whole-embryo culture method was used. For blastocysts with no visible ICM, the whole-embryo culture method was used to establish hESCs via the seeding of the entire blastocyst without its zona pellucida directly on a STO feeder layer. However, trophectodermal overgrowth tends to hinder the expansion of the ICM during the initial steps of hESC derivation. Therefore, the partial-embryo culture method was developed to establish hESCs from blastocysts with smaller ICMs. The surgical isolation of the region containing the ICM with an ultra-fine glass pipette alleviates trophectoderm overgrowth. This method is also applicable to blastocysts with large and distinct ICMs, and the efficiency of this method is comparable to that of the immunosurgical method.     

Effects of an Interferon Inducer, Poly I:C, on Acute Ocular Infection Produced by Intracameral Inoculation of Toxoplasma gondii

A potent interferon inducer, synthetic polyinosinic acid-polycytidylic acid complex (poly I:C), was used prophylactically and therapeutically in experimental ocular Toxoplasma infections in rabbits. Daily intravenous injections of poly I:C alone or when combined with daily subconjunctival injections of the inducer delayed the appearance of conjunctivitis, corneal opacity, and iritis in Toxoplasma-infected eyes provided that the treatment was started 1 day before the infection. When the treatment was begun at the same time or 1 day after the infection, no delay in the production of the ocular lesions was noted. In no case was the treatment curative or completely suppressive.     

Electrochemical properties of suprastructures galvanically coupled to a titanium implant

In recent years, dental implants have been widely used for the aesthetic and functional restoration of edentulous patients. Dental implants and restorative alloys are required with high corrosion resistance. Suprastructures and implants of different compositions in electrical contact may develop galvanic or coupled corrosion problems. In addition to galvanic corrosion, crevice and pitting corrosion may occur in the marginal gap between dental implant assemblies. In this study, gold, silver-palladium, cobalt-chromium, and nickel-chromium suprastructures were used to investigate their galvanic and crevice corrosion characteristics in combination with titanium (Ti) implants. Potentiodynamic and potentiostatic testing were performed in artificial saliva at 37 degrees C. Potentiodynamic testing was carried out at the potential scan rate of 1 mV/s in the range of -600-1600 mV (SCE). Potentiostatic testing was performed with an open-circuit potential and current densities at -250, 0, and 250 mV (SCE) in artificial saliva. After electrochemical testing, surface morphologies and cross-sections were examined using micrographs of the samples. Potentiodynamic test results indicated that suprastructure/Ti implant couples produced passive current densities in the range of 0.5-12 microA/cm(2); Ti abutment/Ti implant and gold/Ti implant couples exhibited relatively low passive current densities; Co-Cr/Ti implant couples the highest. Co-Cr and Ni-Cr/Ti implant couples showed breakdown potentials of 700 and 570 mV (SCE), respectively. The open-circuit potentials of silver, Ti abutment, gold, Ni-Cr, and Co-Cr/Ti implant couples were -93.2 +/- 93.9, -123.7 +/- 58.8, -140.0 +/- 80.6, -223.5 +/- 35.1, and -312.7 +/- 29.8 mV (SCE), respectively, and did not change with immersion time. The couples exhibited cathodic current densities at -250 mV (SCE); in particular, gold and silver alloys showed high cathodic current densities of -3.18 and -6.63 microA/cm(2), respectively. At 250 mV (SCE), Ti abutment/Ti implant couples exhibited a minimum current density of 9.48 x 10(-2) microA/cm(2), but gold, Ni-Cr, Co-Cr, and silver/Ti implant couples exhibited 0.313, 1.27, 5.60, and 8.06 microA/cm(2), respectively. All couples exhibited relatively low current densities at 0 mV (SCE). Photomicrographs after electrochemical testing showed crevice or pitting corrosion in the marginal gap and at the suprastructure surface. Although of the tested samples Co-Cr/Ti implant couples showed the possibility of galvanic corrosion, its degree was not significant. However, it should be borne in mind that galvanic corrosion can accelerate localized corrosion, such as crevice or pitting corrosion.     

Spontaneous disruption of mycotic aneurysm involving innominate artery

We report a case of ruptured mycotic aneurysm involving innominate artery requiring an urgent surgical treatment. A 62-yr-old woman presented with fever and dyspnea. Previously, she was diagnosed with colon cancer and received right hemicolectomy and one cycle of adjuvant chemotherapy. On echocardiogram, pericardial effusion was noted and emergency pericardiocentesis was performed. CT scan revealed aortic aneurysm involving ascending aorta and innominate artery, and thrombi surrounding those structures. Patch repair of the defect in the ascending aorta and ringed Goretex graft to bypass the innominate and ascending aorta were performed. We believe that this is the first case of ruptured mycotic aneurysm involving innominate artery.     

Autologous stem cell transplantation in the treatment of refractory rheumatoid arthritis

The concept of using high-dose immunosuppressive treatment (HDIT) with autologous stem cell transplantation (ASCT) to treat patients with refractory rheumatoid arthritis has been provided by animal studies and anecdotal case reports. Over the past five years, an increasing number of patients with refractory rheumatoid arthritis have received HDIT with ASCT as an adjunct to intense immunosuppression. Here, we present a case of refractory rheumatoid arthritis in a 54-yr-old woman using HDIT with ASCT. Peripheral blood stem cells were mobilized with cyclophosphamide (4 g/m(2)) followed by G-CSF (5 microg/kg/day). Leukapheresis continued daily until the number of harvested progenitor cells reached 2 x 10(6) CD34+ cells/kg after CliniMax CD34+ positive selection. For HDIT, high-dose cyclophosphamide (total dose 200 mg/kg) and antithymocyte globulin (total dose 90 mg/kg) were administered and CD34+ cells were infused 24 hr after HDIT. The patient tolerated the treatment well but experienced an episode of neutropenic fever. She achieved an early dramatic improvement of joint symptoms during therapy. Fifty percent of improvement of rheumatoid arthritis by the American College of Rheumatology (ACR 50) preliminary definition was fulfilled during the 6 months following ASCT. Although further long-term follow-up is required, the patient's activity of arthritis has been stable since receiving HDIT with ASCT.