Hiroshi Takahashi

The retina is an ideal target for gene therapy because of its easy accessibility and limited immunological response. We previously reported that intravitreally injected adeno-associated virus (AAV) vector transduced the inner retina with high efficiency in a rodent model. In large animals, however, the efficiency of retinal transduction was low, because the vitreous and internal limiting membrane (ILM) acted as barriers to transduction. To overcome these barriers in cynomolgus monkeys, we performed vitrectomy (VIT) and ILM peeling before AAV vector injection. Following intravitreal injection of 50 μL triple-mutated self-complementary AAV serotype 2 vector encoding EGFP, transduction efficiency was analyzed. Little expression of GFP was detected in the control and VIT groups, but in the VIT+ILM group, strong GFP expression was detected within the peeled ILM area. To detect potential adverse effects, we monitored the retinas using color fundus photography, optical coherence tomography...

We examined the neuroprotective effects of exogenous brain-derived neurotrophic factor (BDNF), which provides protection to retinal ganglion cells (RGCs) in rodents, in a model of transient intraocular pressure (IOP) elevation using a mutant (triple Y-F) self-complementary adeno-associated virus type 2 vector encoding BDNF (tm-scAAV2-BDNF). The tm-scAAV2-BDNF or control vector encoding green fluorescent protein (GFP; tm-scAAV2-GFP) was intravitreally administered to rats, which were then divided into four groups: control, ischemia/reperfusion (I/R) injury only, I/R injury with tm-scAAV2-GFP, and tm-scAAV2-BDNF. I/R injury was then induced by transiently increasing IOP, after which the rats were euthanized to measure the inner retinal thickness and cell counts in the RGC layer. Intravitreous injection of tm-scAAV2-BDNF resulted in high levels of BDNF expression in the neural retina. Histological analysis showed that the inner retinal thickness and cell numbers in the RGC layer were p...

The transient X-ray binary pulsar A0535+262 was observed with Suzaku on 2005 September 14 when the source was in the declining phase of the August-September minor outburst. The ~103 s X-ray pulse profile was strongly energy dependent, a double peaked profile at soft X-ray energy band (<3 keV) and a single peaked smooth profile at hard X-rays. The width of the primary dip is found to be increasing with energy. The broad-band energy spectrum of the pulsar is well described with a Negative and Positive power-law with EXponential (NPEX) continuum model along with a blackbody component for soft excess. A weak iron K_alpha emission line with an equivalent width ~25 eV was detected in the source spectrum. The blackbody component is found to be pulsating over the pulse phase implying the accretion column and/or the inner edge of the accretion disk may be the possible emission site of the soft excess in A0535+262. The higher value of the column density is believed to be the cause of the secondary dip at the soft X-ray energy band. The iron line equivalent width is found to be constant (within errors) over the pulse phase. However, a sinusoidal type of flux variation of iron emission line, in phase with the hard X-ray flux suggests that the inner accretion disk is the possible emission region of the iron fluorescence line.

ABSTRACT Isotactic polypropylene consisting of uniaxially oriented β-phase lamellae was crystallized in a temperature gradient. The β → α transition was investigated by simultaneous measurements with differential scanning calorimetry (DSC) and X-ray diffraction using synchrotron radiation (SR). To compare the transition mechanism, the β-phase sample was deformed by rolling it along the direction of the crystallization. During rolling, the β-crystal is deformed by interlamellar and interchain slip, which induces c-axis-oriented molecules along the rolling direction. The melting behavior is changed by the rolling deformation. For the as-grown β-crystal, the DSC thermogram has three peaks: the β-melting endotherm at 150°C, an exotherm by recrystallization into the α-form, and the endotherm at 167°C caused by melting of the recrystallized α-form. After the rolling deformation, the β-endotherm is extinguished by the successive exotherm. Simultaneous X-ray measurements reveal that the β → α transition is shifted to a lower temperature and that the recrystallized α-form has a c-axis-orientation caused by the rolling deformation. In the process of the β → α transition, higher-order lamellar structure is developed earlier than formation of the crystalline structure. In this study, the heating phenomena, such as the β → α transition and thickening of the β- and α-lamellae, are consistently explained by a mechanism involving melting and subsequent recrystallization.

In this paper, ultradense wavelength-division multiplexing (WDM) transmission technologies are discussed, and a field demonstration of over-1000-channel ultradense WDM transmission is reported. The generation of an ultradense WDM signal using a supercontinuum multicarrier source that generates more than 1000 carriers and uniform precise channel spacing of 6.25 GHz is presented. The influence of four-wave-mixing generated in the transmission fiber and the requirements placed on the WDM multiplexer and demultiplexer is described. An over-1000-channel ultradense WDM transmission experiment is reported. A 1046 × 2.67-Gbit/s 6.25-GHz-spaced ultradense WDM signal is successfully transmitted over 126 km of field-installed fibers in the test bed of JGN II.

Tau-tubulin kinase 2 (TTBK2) is a Ser/Thr kinase that putatively phosphorylates residues Ser208 and Ser210 (numbered according to a 441-residue human tau isoform) in tau protein. Functional analyses revealed that a recombinant kinase domain (residues 1–331) of human TTBK2 expressed in insect cells with a baculovirus overexpression system retains kinase activity for tau protein. The kinase domain of TTBK2 was crystallized using the hanging-drop vapour-diffusion method. The crystals belong to space group P212121, with unit-cell parameters a = 55.6, b = 113.7, c = 117.3 Å, α = β = γ = 90.0°. Diffraction data were collected to 2.9 Å resolution using synchrotron radiation at BL24XU of SPring-8.

It is demonstrated that the Van der Waals loop appearing in the computer simulations almost vanishes by modifying the shape of the simulation cell even when small size systems are treated. After a long molecular dynamics simulation, it was found that the complete phase separation into vapor and liquid took place in the unstable region. The surface free energy of the interface between the liquid and vapor phases is suggested to be a major reason why the artificial loop has been observed in molecular dynamics simulations. We show that the loop depth diminishes as the box ratio of the simulation cell is enlarged.

Idiopathic interstitial fibrosis (IPF) is a chronic, usually fatal lung disease of unknown etiology. There are few specific therapies for acute exacerbation of IPF and factors predicting the onset or severity of this syndrome are not clearly understood. A neutrophil elastase inhibitor, sivelestat (ONO-5046) has been commercially available in Japan since 2002. This inhibitor has a potent effect in the treatment of ALI/ARDS. To evaluate the outcome of patients with acute exacerbation of IPF treated with sivelestat and estimate prognostic factors, we investigated 10 patients with acute exacerbation of IPF who were intubated and mechanically ventilated. We analyzed the outcome of patients with acute exacerbation of IPF until day 180 and measured the P/F ratio, PEEP levels, the values of peripheral white blood cell number, and C-reactive protein (CRP) on day 0, 3, 7 after admission. Serum KL-6 and surfactant protein D (SP-D) concentration on day 0 were also analyzed. All patients were treated with sivelestat and methylprednisolone (mPSL) pulse therapy for 3 days from day 0 and maintenance therapy with prednisone (0.5 mg/kg/day) were continued. Of the 10 patients. 4 patients had survived (40%) and 6 patients had died (60%) at day 180 from the onset of acute exacerbation of IPF. In survivors, P/F ratio, PEEP levels, and CRP values significantly improved on day 7 (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05). Serum KL-6 and SP-D were lower in survivors on day 0 (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05). Taken together, serum KL-6 and SP-D may prove valuable as biochemical markers of prognosis in acute exacerbation of IPF. Sivelestat may have potential in the treatment of acute exacerbation of IPF.

The objective of this study was to evaluate the efficacy and toxicity of carboplatin plus weekly paclitaxel as first-line therapy in patients with advanced non-small cell lung cancer (NSCLC). Forty-nine patients were analyzed retrospectively. Every 4 weeks patients received 70 mg/m(2)paclitaxel on days 1, 8, and 15, and AUC 5-6 carboplatin on day 1. A median of four cycles (range, 1-7) was administered. Twenty-four patients had a partial response, and the overall response rate was 48.9%. The median survival time was 12.8 months and the 1-year survival was 50.7%. Overall toxicities were mild. The most common toxicity was neutropenia, grade 3/4 in 32% of the patients. Grade 3/4 hematologic toxicities included anemia (16%) and thrombocytopenia (8%). Grade 3/4 non-hematologic toxicities included febrile neutropenia (2%), pneumonia (10%) and interstitial pneumonia (2%). Grade 2 peripheral neuropathy was seen in one patient (2%). These results demonstrate that this regimen is an active and tolerable treatment for patients with advanced NSCLC. It is suggested that this weekly regimen should be considered as one of the standard therapies for future chemotherapy in advanced NSCLC.

We investigated the significance and the usefulness of monitoring plasma voriconazole levels in patients with chronic necrotizing pulmonary aspergillosis associated with underlying chronic respiratory diseases. The average trough level was 2.2 microg/ml and there was no correlation between trough levels and voriconazole doses. Orally administered drug showed no significant difference in trough or peak levels compared with parenteral injection. Six cases with visual adverse events had significantly higher nadirs compared to those without visual disturbance. All three cases who discontinued the drug due to liver dysfunction had plasma trough levels higher than 4.0 microg/ml. Those who failed to respond to the treatment had trough levels lower than 1.4 microg/ml or peak levels lower than 2.8 microg/ml, while some cases with plasma level lower than those levels responded well. Since plasma voriconazole level has a large inter-patient variability, drug monitoring may be beneficial to evaluate the drug efficacy and safety in each individual.

This study was retrospectively conducted to clarify the clinical features of pneumothorax in patients with active nontuberculous mycobacterial (NTM) lung disease. The patients were 9 men and 7 women, with a median age of 70. Pathogenic mycobacteria were 12 Mycobacterium avium complex, 2 Mycobacterium kansasii, and 2 Mycobacterium fortuitum. More than one lung field was affected by NTM disease in 11 patients, whereas 4 patients were given a diagnosis of mild NTM disease only at the onset of pneumothorax. Five patients recovered with rest only, 4 with thoracic drainage, 4 needed surgery, and 2 developed chronic pneumothorax. Five patients experienced more than one recurrence of pneumothorax during treatment for NTM disease. The rate of the complication of pneumothorax in patients with NTM lung disease was estimated at around 2.3%. Pneumothorax associated with NTM lung disease is not rare and is sometimes difficult to control.

A 71-year-old man presented with a thin-walled cavity in his left lung in November 2006. A previous chest CT in 2003 showed a small thin-walled cavity in his left lingula. Although no obvious change was observed in 2004, the cavity increased its size from 11mm to 14mm in diameter and the wall became thicker in June 2006. On the first visit to our hospital in November 2006, the diameter of the cavity was 30mm and some part of the wall was thinner than on the previous CT. The patient developed pneumothorax one month later and underwent segmentectomy of the left lingula after unsuccessful thoracic drainage. Poorly differentiated adenocarcinoma was identified in both the pleura and the inner wall around the cavity. Lung adenocarcinoma with gradual enlargement of a thin-walled cavity causing pneumothorax has never been reported before. We report here the natural course of lung adenocarcinoma with a thin-walled cavity.

Flashing experiments on superheated liquids under rapid depressurization were carried out using the JAMIC drop tower in order to reexamine the existing thermodynamic nucleation theory and to observe bubble growth and collapse in microgravity. Flashing bubble behavior was studied by visualization, and temperature and pressure transients within the vessel were measured.

Haemophilus influenzae, a major respiratory tract pathogen, is becoming increasingly resistant to beta-lactam antibiotics. Studying annual trends in antibiotic susceptibility and genetic patterns of H. influenzae beta-lactam resistance, we isolated 122 strains from the adult respiratory tract in 2007, determined MIC for different antibiotics, and analyzed TEM-1 beta-lactamase resistant genes and ftsI encoding PBP3 mutation compared to results in 2005 and 2007. We found that ABPC-susceptible strains with MIC &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;1 microg/mL (BLNAS) accounted for 71.0%, ABPC-resistant strains with MIC exceeding 2 microg/mL without beta-lactamase activity (BLNAR) for 25.3%, and beta-lactamase-positive strains (BLP) for 3.7%. The BLNAS ratio showed no significant change from 2002 and 2005. The BLP ratio decreased from those in 2002 and 2005. Genetic studies of resistant genes showed that gBLNAS with no resistant genes had increased in the last five years. The ratio of all strains with PBP3 mutation (gBLNAR and gLow-BLNAR) remained constant from 2002 to 2007. The proportion of gBLNAR with two PBP3 mutations had increased, however, while gLow-BLNAR with one mutation had decreased. LVFX showed constant strong antimicrobial potency for all mutation groups. Among beta-lactam antibiotics, the lowest MIC90 was observed in parenteral CTRX and oral CDTR-PI use. Although a new MIC peak generated by gBLNAR became obvious in the ABPC and CDTR-PI MIC distribution, the MIC of the new peak was still low enough to treat with high doses of those two antibiotics.

Cyanamide, an aversive agent widely used in Japan, is known to induce various degrees of hepatic lesion with ground-glass inclusion bodies. When cyanamide-treated alcoholics relapse into drinking, more severe inflammation develops in the liver. However, it is controversial whether progressive hepatic lesions develop in complete abstainers as a result of long-term cyanamide treatment. Case 1: A 53-year-old male alcoholic received cyanamide treatment for 4.5 months and completely abstained without cyanamide treatment for 6 years. A liver biopsy shortly after abstinence showed extensive pericellular fibrosis, but a biopsy after 6 years showed very mild fibrosis. Case 2: A 43-year-old male alcoholic remained completely abstinent with cyanamide treatment for 5 years and complained of general fatigue. His serum transaminases were slightly elevated and hepatic hyperechogenicity was observed on ultrasonography. Only mild pericellular fibrosis was present in the liver biopsy specimen obtained shortly after abstinence, but after 5 years the second liver biopsy showed that thin septum-like fibrosis that formed portal-to-portal and portal-to-central linkage had developed and ground-glass hepatocytes had emerged extensively. Case 3: A 29-year-old female alcoholic complained of general fatigue and a slight fever after 1.5 years of abstinence with cyanamide treatment. Slight elevation of serum transaminases and hepatic hyperechogenicity were observed. The liver biopsy showed extensive ground-glass hepatocytes and thin septum-like fibrosis that formed portal-to-portal linkage. Case 4: A 61-year-old male alcoholic who remained completely abstinent while taking cyanamide for 3 years showed slight elevation of serum transaminases. Liver biopsy showed extensive ground-glass hepatocytes and extension of thin septum-like fibers from portal tract to the lobule. Ultrasonography revealed hepatic hyperechogenicity. In some abstainers who take cyanamide for several years, thin septum-like liver fibrosis progresses along with the emergence of ground-glass hepatocytes. Hepatic hyperechogenicity on ultrasonography and slight elevation of serum transaminases might erroneously lead to a diagnosis of hepatic steatosis without liver histology.

Distigmine bromide is a cholinesterase inhibitor widely used for the treatment of hypotonic neurogenic bladder. However, this drug is also known to cause cholinergic crisis, a rare but serious adverse reaction. Cholinergic crisis is an excessive amount of acetylcholine due to the systemic inhibition of cholinesterase activity, characterized by parasympathetic symptoms such as sweating, salivation, miosis, bradycardia, diarrhea and circulatory and respiratory failure. The incidence of cholinergic crisis has been estimated at approximately 0.2%, and the majority of the patients are elderly with underlying conditions such as cerebrovascular disease. Since 2004, we have encountered 5 cases of acute respiratory failure associated with cholinergic crisis induced by the administration of a normal oral dose of distigmine. We present these cases here and review an additional 23 cases from the literature in Japan. In these 28 cases, mechanical ventilation was required for 57%, with a mean duration of 5.1 days and a mortality rate of 11%. Pneumonia was observed in half of the cases in the acute phase, and relapse due to the readministration of distigmine was reported in 20% of cases. It is important to remember that cholinergic crisis in the elderly is often misdiagnosed and is occasionally treated as simple aspiration pneumonia.

We determined beta-lactamase activity and antimicrobial susceptibility of 556 strains consisting of 10 species isolated in four medical institutions and one microbiological laboratory of Miyagi Prefecture in Japan between December in 1999 and February in 2000. beta-Lactamase determined by nitrocefin method was positive in 68% of S. aureus, in 15% of H. influenzae and in 100% of M. catarrhalis. Penicillinase/cephalosporinase determined by acidometry was positive in 9%/10% of E. coli, in 17%/2% of K. pneumoniae, in 16%/58% of E. cloacae, in 43%/78% of S. marcescens, and in 4%/32% of P. aeruginosa, respectively. Of a total of 298 strains of Enterobacteriaceae and P. aeruginosa, 25 strains (14 strains of E. coli, 10 strains of K. pneumoniae and one strain of S. marcescens) produced class A beta-lactamase, two strains of E. cloacae produced class B beta-lactamase, and 12 strains (one strain of E. coli, four strains of E. cloacae, six strains of S. marcescens and one strain of P. aeruginosa) produced class C beta-lactamase. According to NCCLS standard, three strains (one strain of E. coli and two strains of K. pneumoniae) of ESBL-positive microbes were detected. beta-Lactamase-negative ampicillin-resistant (BLNAR) strains of H. influenzae were found in 10/40 (25.0%) of the strains tested.

We studied the clinical features and efficacy of home noninvasive positive pressure ventilation (NPPV) therapy in 80 patients to ascertain its indications and problems. The causes of chronic respiratory failure were restrictive thoracic diseases of post-tuberculosis sequelae (40 cases) and kyphoscoliosis (9 cases), COPD (8 cases), bronchiectasis (7 cases), and interstitial pneumonia (4 cases). One year survival rate of the patients with post-tuberculosis sequelae was 76% and most of the patients who started NPPV at their acute exacerbation died within several months. About half of the patients of COPD improved their quality of life (QOL) through NPPV. However, their survival rate 3 months later was only 69%. More than half of the patients with bronchiectasis felt that their QOL was improved by NPPV. Most of the patients with interstitial pneumonia died within 3 months indicating that NPPV is less useful for improving QOL of interstitial pneumonia PaCO2, after home NPPV, decreased significantly in the responder group (70.0 +/- 15.4 vs. 57.6 +/- 10.7[SD]Torr, p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05), while PaCO2 in the non-responder group was unchanged (65.4 +/- 12.1 vs. 64.2 - 10.4 [SD] Torr). Body Mass Index (BMI) in the responder group tended to be higher than in the non-responder group. In conclusion, the restrictive thoracic diseases with post-tuberculosis sequelae and kyphoscoliosis are a good indication for NPPV and the therapy is also useful for patients with bronchiectasis who can dispose of their sputum by themselves. Home NPPV is suitable for patients whose PaCO2 decreases through NPPV and whose BMI is relatively high. QOL of interstitial pneumonia barely improves through NPPV, because interstitial pneumonia with hypercapnia is at the terminal stage.

To evaluate effectiveness of pneumococcal polysaccharide vaccine, we recommended 1378 outpatients aged over 60 with chronic respiratory diseases to be vaccinated from August to October 2002, and 647 patients were vaccinated from August to November 2002. In the 1229 patients without respiratory failure, the incidence of antimicrobial treatment for bacterial respiratory infections in 547 vaccinated patients significantly decreased from 7.9% in the 2001/02 winter season to 5.7% in the 2002/03 winter season, although that in the 682 unvaccinated patients increased from 3.8% to 5.7%. The incidence of antimicrobial treatment for bacterial respiratory infections in 229 vaccinated patients with pneumococcal and influenza vaccines together significantly decreased from 10.5% in the 2001/02 winter season to 5.2% in 2002/03 winter season although that in 110 subjects vaccinated with influenza vaccine only increased from 2.7% to 7.2%. These findings suggest the effectiveness of the pneumococcal polysaccharide vaccine for the prevention of bacterial respiratory infections and the additive effectiveness of pneumococcal and influenza vaccines together.