Successful control of an outbreak of GES-5 extended-spectrum ß-lactamase-producing Pseudomonas aeruginosa in a long-term care facility in Japan.

BACKGROUND: Little is known about multidrug-resistant Pseudomonas aeruginosa (MDRP) outbreaks in long-term care facilities (LTCFs). AIM: To describe an MDRP outbreak in an LTCF and to clarify risk factors for MDRP acquisition. METHODS: Patients who were positive for MDRP at an LTCF from January 2013 to January 2014 were analysed. A descriptive analysis, a case-control study, and a microbiological analysis were performed. FINDINGS: A total of 23 MDRP cases were identified, 16 of which were confirmed in sputum samples. Healthcare workers were observed violating hand hygiene procedures when performing oral, wound, and genital care. Nasogastric tube and oxygen mask use was associated with MDRP acquisition in the respiratory tract, which might have been confounded by poor hand hygiene. Sharing unhygienic devices, such as portable oral suction devices for oral care, and washing bottles and ointments for wound and genital care with inadequate disinfection could explain the transmission of MDRP in some cases. Isolates from 11 patients were found to be indistinguishable or closely related by pulsed-field gel electrophoresis and harbouring the blaGES-5 gene. Subsequent enhanced infection control measures were supported by nearby hospitals and a local public health centre. No additional cases were identified for a year after the last case occurred in January 2014. CONCLUSION: An outbreak of MDRP with an antimicrobial resistance gene, blaGES-5, occurred in a Japanese LTCF. It was successfully controlled by enhanced infection control measures, which neighbouring hospitals and a local public health centre supported.

Evaluation of effective energy using radiochromic film and a step-shaped aluminum filter.

Although the half-value layer (HVL) is one of the important parameters for quality assurance (QA) and quality control (QC), constant monitoring has not been performed because measurements using an ionization chamber (IC) are time-consuming and complicated. To solve these problems, a method using radiochromic film and step-shaped aluminum (Al) filters has been developed. To this end, GAFCHROMIC EBT2 dosimetry film (GAF-EBT2), which shows only slight energy dependency errors in comparison with GAFCHROMIC XR TYPE-R (GAF-R) and other radiochromic films, has been used. The measurement X-ray tube voltages were 120, 100, and 80 kV. GAF-EBT2 was scanned using a flat-bed scanner before and after exposure. To remove the non-uniformity error caused by image acquisition of the flat-bed scanner, the scanning image of the GAF-EBT2 before exposure was subtracted after exposure. HVL was evaluated using the density attenuation ratio. The effective energies obtained using HVLs of GAF-EBT2, GAF-R, and an IC dosimeter were compared. Effective energies with X-ray tube voltages of 120, 100, and 80 kV using GAF-EBT2 were 40.6, 36.0, and 32.9 keV, respectively. The difference ratios of the effective energies using GAF-EBT2 and the IC were 5.0%, 0.9%, and 2.7%, respectively. GAF-EBT2 and GAF-R proved to be capable of measuring effective energy with comparable precision. However, in HVL measurements of devices operating in the high-energy range (X-ray CT, radiotherapy machines, and so on), GAF-EBT2 was found to offer higher measurement precision than GAF-R, because it shows only a slight energy dependency.

Half-value layer measurement: simple process method using radiochromic film.

Although the half-value layer (HVL) is one of the important parameters for QA and QC, constant monitoring has not been performed because the measurements using an ionization chamber (IC) are time-consuming and complicated. To solve these problems, the use of radiochromic film (GAFCHROMIC XR TYPE R: GAF-R) with step-shaped aluminum (Al) filters, referred to herein as the simple process method, has been developed. The measurement X-ray tube voltages were 120 kV, 100 kV, and 80 kV. The Al filter area, the full exposure area, and the unexposed area were set on the GAF-R so as to obtain correct data. The HVL was evaluated using the density attenuation ratio. The HVLs obtained using the GAF-R and an 1C dosimeter were compared. HVLs with X-ray tube voltages of 120 kV, 100 kV, and 80 kV using the GAF-R were 4.10 mm, 3.55 mm and 2.97 mm, respectively. The difference ratios of the HVLs using the GAF-R and the IC were 1.2%, 7.6%, and 10.0%, respectively. The HVL at 120 kV can be routinely and quickly measured using the simple process method. Therefore, an IC dosimeter is not needed for HVL measurements for QA and QC. However, the HVL measurements of low energy (100 kV and 80 kV) need attention.

Evaluation of effective energy for QA and QC: measurement of half-value layer using radiochromic film density.

The effective energy of diagnostic X-rays is important for quality assurance (QA) and quality control (QC). However, the half-value layer (HVL), which is necessary to evaluate the effective energy, is not ubiquitously monitored because ionization-chamber dosimetry is time-consuming and complicated. To verify the applicability of GAFCHROMIC XR type R (GAF-R) film for HVL measurement as an alternative to monitoring with an ionization chamber, a single-strip method for measuring the HVL has been evaluated. Calibration curves of absorbed dose versus film density were generated using this single-strip method with GAF-R film, and the coefficient of determination (r2) of the straight-line approximation was evaluated. The HVLs (effective energies) estimated using the GAF-R film and an ionization chamber were compared. The coefficient of determination (r2) of the straight-line approximation obtained with the GAF-R film was more than 0.99. The effective energies (HVLs) evaluated using the GAF-R film and the ionization chamber were 43.25 keV (5.10 mm) and 39.86 keV (4.45 mm), respectively. The difference in the effective energies determined by the two methods was thus 8.5%. These results suggest that GAF-R might be used to evaluate the effective energy from the film-density growth without the need for ionization-chamber measurements.

[Subtraction image for dynamic liver MRI using free breath-hold at functional residual capacity: a clinical trial].

PURPOSE: Dynamic liver MRI images have been obtained under expiration breath holding (BH). However, problems with obtaining reproducible liver positions often observed. This study investigated ways to improve the reproducibility of liver position on dynamic liver MRI. MATERIALS AND METHODS: After giving informed consent, 60 patients (32 males and 28 females, ages 33-85, median age 69) were examined by liver dynamic MRI under two types of BH. The BH phases were voluntary expiration (VE) phase without any explanations and functional residual capacity (FRC) phase after careful explanation was provided. Plain images, arterial phase images, portal phase images and parenchymal phase images were obtained. For statistical evaluation of reproducibility, the area of the 2nd or 3rd images from top of the liver was measured in each phase using a threshold value of half maximum. Misregistration areas were calculated by finding the remainder of the liver area in the plain-arterial (Pl-A) phase, arterial-portal (A-Po) phase, plain-parenchymal (Pl-Pa) phase. Contingency table analysis was done due to the misregistration was occurred or not. RESULTS: Misregistration of liver image on the VE and the FRC of three phase types were statistical significant on the Pl-A (p < 0.01), on the A-Po (p < 0.01) and on the Pl-Pa (p < 0.05), respectively. CONCLUSION: The FRC phase following careful explanation of the BH provided significantly improved reproducibility of liver position on dynamic liver MRI. Therefore, precise subtraction images could be obtained for routine clinical examinations without slice matching.

Dose distribution in pediatric CT head examination using a new phantom with radiochromic film.

Performing a detailed dose measurement is important to keep radiation doses during computed tomography (CT) examinations as low as reasonably achievable. To estimate in detail the dose distribution during pediatric CT examination of the head, a sheet roll CT dosimetry phantom (SRCT-P) with radiochromic film (RF) was developed. The dose distributions in the SRCT-P (diameters of 6 [premature baby], 10 [neonate], and 14 [infant] cm) were evaluated. The SRCT-Ps were made by rolling up flexible acrylic sheets (1.1 g/cm3). RFs were positioned every 5 mm along the radius at each SRCT-P, starting at 10 mm (center) and ending on the surface. The dose distribution along the z-axis at the center or on the surface showed a flat or wave pattern, respectively. When the mean surface dose at 10 cm diameter was taken as 100%, the mean surface doses at 6 or 14 cm diameters were 105 or 96%, respectively, and the mean center doses at 6, 10, and 14 cm were 109, 99, and 74%, respectively. The maximum-minimum doses and dose distribution of a CT examination can be measured separately by using the SRCT-P with RF.

Computed tomography phantom for radiochromic film dosimetry.

To evaluate in detail the dose distribution during computed tomography (CT), a sheet roll CT dosimetry phantom (SRCT-P) with a radiochromic film (RF) was experimentally developed. The SRCT-P was made by rolling up a vinyl chloride sheet in a cylindrical shape to arbitrarily select the SRCT-P diameter, dose measurement position, and depth. The SRCT-P centre core consisted of a plastic hose in which a 10 mm acrylic bar with a RF was inserted. To determine the availability of the SRCT-P, the surface and centre doses (at a 5 mm radius) at each SRCT-P diameter (6-16 cm; every 2 cm) were measured. The ratios of the centre-to-surface doses (D(centre)/D(surface)) systematically increased, from 80 to 111%, for decreasing SRCT-P diameters, between 16 and 6 cm, respectively. The centre dose approached the surface dose as the SRCT-P diameter decreased. To use a RF for a CT dose measurement, further detailed research and analysis is necessary. However, this study has shown that a SRCT-P is useful and beneficial for the measurement of the dose distribution during a CT examination.

[Surgical treatment of thymic carcinomas].

Five cases of surgically treated thymic carcinoma are reported. The patients (4 men and a woman) ranged in age from 46 to 76 years old with a mean of 64.6. Four patients were asymptomatic and an abnormal shadow on X-ray films was noted. One remaining patient suffered from hoarseness. One patient had stage II disease and the others had stage III. Surgical tumor resection was performed in all cases. Only 1 patient among the 5 underwent a successful complete resection. Histological examinations of the resected specimens revealed squamous cell carcinoma of thymus. Four specimens were poorly differentiated and 1 is moderately differentiated carcinoma. All patients received radiation therapy post operatively. Three patients are alive without any recurrence 6, 8 and 109 months after the surgery. Thymic carcinomas are frequently invasive or metastatic at the time of diagnosis. But poorly differentiated group, in squamous cell carcinoma, mucoepidermoid carcinoma and besaloid carcinoma, are characterized by a low incidence of local recurrence and distant metastasis. They also have a good sensitivity for the radiation. Therefore complete surgical resection combined with postoperative radiation therapy should be a choice in treating thymic carcinomas. We considered that complete resection and postoperative radiation therapy is a curative therapy for thymic carcinomas.

Identification of protein kinase Calpha as an essential, but not sufficient, cytosolic factor for Ca2+-induced alpha- and dense-core granule secretion in platelets.

Upon activation, platelets release many active substances. Here, we have analyzed the mechanism governing Ca(2+)-induced secretion of von Willebrand factor stored in alpha-granules and 5-hydroxytryptamine in dense-core granules in permeabilized human platelets. Both secretions were dependent on ATP and cytosol. An essential factor for both granule secretions was purified from rat brain cytosol and identified to be protein kinase Calpha (PKCalpha) by partial amino acid sequencing. Purified PKCalpha efficiently stimulated both secretions in the presence of cytosol, whereas PKCalpha alone did not support the secretion of either type of granules, suggesting that PKCalpha is not a sufficient factor. Finally, in human platelet cytosol fractionated by a gel filtration column, the stimulatory activity for dense-core granule secretion paralleled with the concentration of PKC, suggesting that PKC could also be such a stimulatory factor in platelet cytosol. Thus, we identified PKCalpha as an essential, but not sufficient, cytosolic factor for the Ca(2+)-induced secretions of both alpha- and dense-core granules in platelets.

Novel splice variants of PACAP gene in mouse cerebellar granule cells.

The mouse pituitary adenylate cyclase activating polypeptide (PACAP) gene consists of two alternative exons (1A and 1B) and four exons including the coding region. By the usage of alternative exons, three transcripts can be produced. In this report, however, we found that at least five preproPACAP transcripts, including two novel types, were produced in mouse cerebellar granule cells (CGCs) and their expression was induced by the Ca2+ influx through L-type voltage-dependent Ca2+ channels. The splicing occurred between the 3'-donor site of exon 1A and the two novel internal 5'-acceptor sites of exon 1B. These findings suggest that preproPACAP transcripts are produced by the complicated usage of the 5'-untranslated region in an activity-dependent manner.

A new type of endo-xyloglucan transferase devoted to xyloglucan hydrolysis in the cell wall of azuki bean epicotyls.

A new type of xyloglucan-degrading enzyme was isolated from the cell wall of azuki bean (Vigna angularis Ohwi et Ohashi cv. Takara) epicotyls and its characteristics were determined. The enzyme was purified to apparent homogeneity by Concanavalin A (Con A)-Sepharose, cation exchange, and gel filtration columns from a cell wall protein fraction extracted with 1 M sodium chloride. The purified enzyme gave a single protein band of 33 kDa on SDS-PAGE. The enzyme specifically cleaved xyloglucans and showed maximum activity at pH 5.0 when assayed by the iodine-staining method. An increase in reducing power in xyloglucan solution was clearly detected after treatment with the purified enzyme. Xyloglucans with molecular masses of 500 and 25 kDa were gradually hydrolyzed to 5 kDa for 96 h without production of any oligo- or monosaccharide with the purified enzyme. The purified enzyme did not show an endo-type transglycosylation reaction, even in the presence of xyloglucan oligosaccharides. Partial amino acid sequences of the enzyme shared an identity with endo-xyloglucan transferase (EXGT) family, especially with xyloglucan endotransglycosylase (XET) from nasturtium. These results suggest that the enzyme is a new member of EXGT devoted solely to xyloglucan hydrolysis.

Involvement of endogenous PACAP expression in the activity-dependent survival of mouse cerebellar granule cells.

Membrane depolarization causes Ca2+ influx through L-type voltage-dependent calcium channels (L-VDCC), which promotes the activity-dependent survival of mouse cerebellar granule cells (CGCs). Although exogenously added pituitary adenylate cyclase activating polypeptide (PACAP) is effective in promoting the survival of CGCs, it is unknown whether PACAP is synthesized in CGCs and involved in the activity-dependent survival of CGCs. In this study, we found that the PACAP gene was activated in depolarized CGCs cultured at 25 mM KCl (high K+), independently of de novo protein synthesis. In addition, the PACAP immunoreactivity increased through the activation of L-VDCC in depolarized CGCs, indicating that PACAP is concomitantly produced with PACAP mRNA in an activity-dependent manner. Exogenously added PACAP attenuated the apoptosis of CGCs through a specific interaction with PACAP receptors. Furthermore, a PACAP receptor antagonist, PACAP(6-38), reduced the survival of CGCs at high K+. These findings indicate that endogenous PACAP production induced by Ca2+ signals exerts a survival effect on CGCs via PACAP receptors, which, at least in part, accounts for the activity-dependent survival of CGCs.

Small GTPase Rho regulates thrombin-induced platelet aggregation.

Platelets play essential roles in hemostasis and thrombosis by aggregating with each other. However, the molecular mechanism governing platelet aggregation is not yet fully understood. Here, we established an assay system using platelets permeabilized with streptolysin-O to analyze mechanism of the thrombin-induced aggregation, focusing upon a controversial issue in the field whether small GTPase Rho regulates the aggregation. Incubation of the permeabilized platelets with Rho GDP-dissociation inhibitor, an inhibitory regulator for Rho family GTPases, extracted Rho family proteins extensively from the plasma and intracellular membranes, and inhibited the thrombin-induced aggregation. Incubation of the permeabilized platelets with botulinum exoenzyme C3, which specifically inhibits Rho function by ADP-ribosylating it, abolished the thrombin-induced aggregation. Thus, Rho is involved in thrombin-induced aggregation of platelets.

[Studies on kinetic viscoelasticity of slow muscle fibers--2. Dynamic stiffness changes studied by quick release method and frequency response method].

PURPOSE: To evaluate the mechanical properties of slow fibers and fast fibers which make up the extraocular muscles in rabbits. SUBJECTS AND METHOD: I studied the contractile properties and viscoelastic properties of the superior rectus muscle(SR) and the retractor bulbi muscle (RB) of rabbits using the quick release method and the frequency response method. RESULTS: In the quick release method, isometric tension transients were slower in the SR than in the RB. In the frequency response method, the muscle length is perturbed sinusoidally to measure kinetic viscoelasticity. In resting muscle, dynamic stiffness was not changed with increasing frequency. In contracted muscle, dynamic stiffness was increased with increasing frequency, and the rate of increase was greater in the SR than in the RB. CONCLUSION: It was concluded that the viscoelasticity of the activated cross bridge is greater in slow fibers than in fast fibers.

[Examination of eye position after strabismus surgery in Kawasaki Medical School Hospital].

PURPOSE: To examine the factors which affect changes in the position of the eyes following the strabismus surgery. METHODS: The postoperative position of 240 eyes with concomitant strabismus was examined. The factors which seemed to have an effect on the position of the eyes were enumerated, and a logistic regression analysis of the results was done. RESULTS: In comparison with esotropia, in exotropia the eyes showed a tendency to return gradually to a normal position. There was no remarkable improvement in stereoscopic vision following strabismus surgery. Based on the standards of the Japanese Association of Strabismus and Amblyopia [the cure standard for strabismus], the results of strabismus surgery were as follows: Grade 1(cosmetically satisfactory) was mainly observed for esotropia, and Grade 3(good) for exotropia. The factors which had a significant effect on the postoperative position of the eyes were as follows: esotropia amblyopia, muscle movement, and operative methods; exotropia: retinal correspondence, operative methods, and the necessity of inferior oblique muscle surgery. CONCLUSIONS: There was little correction of the position of the eyes. In individual cases, the factors which affect position of eyes were examined, and it was considered necessary to increase the amount of correction. Age is a factor which has an effect, but the effect on esotropia and exotropia is small.

Molecular dissection of alpha- and dense-core granule secretion of platelets.

Upon activation, platelets release many active substances stored in alpha- and dense-core granules. However, the molecular mechanisms governing the regulated exocytosis are not yet fully understood. We have established an assay system using streptolysin-O-permeabilized platelets to analyze the Ca2+-induced secretions of von Willebrand factor stored in alpha-granules and [3H]5-hydroxytryptamine (5-HT) in dense-core granules. Using the assay, we found that small GTPase Rab4 regulates alpha-, but not dense-core, granule secretion in platelets. Furthermore, we purified a cytosolic essential protein and currently are analyzing its function.

Repression of activity-dependent c-fos and brain-derived neurotrophic factor mRNA expression by pyrethroid insecticides accompanying a decrease in Ca(2+) influx into neurons.

Permethrin, a type I pyrethroid insecticide, is known to affect sodium channels of neurons and prolong sodium currents. On the other hand, the expression of brain-derived neurotrophic factor (BDNF) and c-fos genes is activated through Ca(2+) influx into neurons, in an activity-dependent manner. In this study, therefore, we investigated whether permethrin influenced the Ca(2+) signal-induced expression of these genes. In primary culture of mouse cerebellar granule cells (CGCs), stimulation with veratridine, a potent agonist for sodium channels, which causes membrane depolarization in neurons, induced c-fos and BDNF mRNA expression accompanying the Ca(2+) influx into neurons. Pretreatment with permethrin at doses nontoxic to CGCs repressed the induction of these genes dose dependently, with trans-permethrin more potent than cis-permethrin. Consistent with this, the increase in Ca(2+) influx caused by veratridine was repressed by permethrin. The membrane depolarization induced by elevating the potassium (K(+)) concentration in medium (high K(+)) caused the activation of c-fos and BDNF genes, which was also repressed by permethrin. Immunoblotting analysis of c-Fos and a gel-mobility assay of AP-1 DNA-binding activity supported the decrease in c-Fos synthesis in permethrin-treated CGCs. The type II pyrethroid cypermethrin also affected the expression of these genes but less effectively than permethrin. Thus, pyrethroids inhibit the activity-dependent gene expression in neurons.

Small GTPase Rab4 regulates Ca2+-induced alpha-granule secretion in platelets.

Upon activation, platelets release many active substances stored in alpha- and dense-core granules. However, the molecular mechanisms governing regulated exocytosis are not yet fully understood. Here, we have established an assay system using permeabilized platelets to analyze the Ca(2+)-induced exocytosis of both types of granules, focusing on RabGTPases. Incubation with Rab GDP dissociation inhibitor, an inhibitory regulator of RabGTPases, reduced membrane-bound RabGTPases extensively, and caused strong inhibition of the Ca(2+)-induced secretion of von Willebrand factor (vWF) stored in alpha-granules, but not that of [(3)H]5-hydroxytryptamine (5-HT) in dense-core granules. Specifically, Rab4 co-fractionated with vWF and P-selectin (an alpha-granule marker) upon separation of platelet organelles by density gradient centrifugation. Incubation of the permeabilized platelets with cell extracts expressing the dominant negative mutant of His-tagged Rab4S22N, but not with those of similar mutant His-Rab3BT36N, inhibited the vWF secretion, whereas neither of the cell extracts affected the [(3)H]5-HT secretion. Importantly, the inhibition of vWF secretion was rescued by depleting the cell extracts of the His-Rab4S22N with nickel beads. Thus, in platelets, the regulatory mechanisms governing alpha- and dense-core granule secretions are distinct, and Rab4 is an essential regulator of the Ca(2+)-induced exocytosis of alpha-granules.