Generation of ozone during irradiation using medical linear accelerators: an experimental study.

BACKGROUND: Some patients have noted a foul odor during radiation therapy sessions, but the cause of the odor remains unknown. Since we suspected that this phenomenon is due to ozone generated by ionizing radiation, this experimental study measured ozone concentrations in the treatment room and in a coiled polyvinyl chloride (PVC) tube placed within the radiation field. METHODS: We measured ozone concentrations using an ultraviolet absorption method and an ozone monitor. A PVC tube (inner diameter 7 mm, outer diameter 10 mm) was used to mimic the environment of the nasal cavity. The tube (790 cm) was coiled and set between two 4-cm-thick (for X-rays) or 2-cm-thick (for electron beams) water-equivalent solid phantoms. The sampling tube of the ozone monitor was inserted into the PVC tube, and the joint was sealed to prevent environmental air contamination. To measure ozone concentrations in the atmosphere, the sampling tube supplied with the unit was used. A linac was used on a full-sized treatment field (40 cm × 40 cm at a source-to-axis distance of 100 cm). The effect of an electron beam on ozone concentrations was also evaluated with a full-sized treatment field (40 cm × 40 cm at a source-to-surface distance of 100 cm). RESULTS: Ozone levels in the treatment room were undetectable before the start of daily treatment but reached 0.008 parts per million (ppm) or more at 1 h after the start of treatment. Concentrations then remained nearly constant at 0.010-0.015 ppm throughout the day. The maximum ozone concentration in the PVC tube was only 0.006 ppm, even when it was irradiated at 2400 monitor units/min. Depending on the X-ray dose rate, the concentration increased to a maximum of 0.010 ppm with oxygen flowing into the other end of the tube at 1.5 L/min. Ozone concentrations in the PVC tube did not differ significantly between X-ray and electron-beam irradiation. CONCLUSIONS: Only traces of ozone were found in the PVC tube that was used to mimic the nasal passages during radiation, these concentrations were too low for human perception. However, ozone concentrations did reach potentially detectable levels in the treatment room.

Expression of serine protease inhibitors in epidermal keratinocytes is increased by calcium but not 1,25-dihydroxyvitamin D3 or retinoic acid.

BACKGROUND: In human skin, the serine proteases kallikrein-related peptidase (KLK)5 and KLK7 degrade corneodesmosome proteins, leading to desquamation. Serine protease activity of the skin is tightly regulated by the interplay between such proteases and serine protease inhibitors, including lymphoepithelial Kazal-type related inhibitor (LEKTI), encoded by SPINK5; secretory leucocyte peptidase inhibitor (SLPI); and elafin. Expression of KLK5 and KLK7 is controlled and upregulated by stimulants such as calcium, 1,25-dihydroxyvitamin D3 [1,25(OH)2 VD3 ] and retinoic acid (RA). OBJECTIVES: To understand the effect of calcium, 1,25(OH)2 VD3 and RA on the expression of serine protease inhibitors in epidermal keratinocytes. METHODS: We stimulated normal human epidermal keratinocytes (NHEKs) with high calcium, 1,25(OH)2 VD3 or RA, and then analysed the expression of serine protease inhibitors using quantitative real-time polymerase chain reaction, enzyme-linked immunosorbent assay and immunocytofluorescence. We also analysed trypsin- and chymotrypsin-like serine protease activities in stimulated NHEKs. RESULTS: High calcium, but not 1,25(OH)2 VD3 or RA, significantly induced the expression of LEKTI, SLPI and elafin at both transcript and protein levels in NHEKs. These inductions were time- and dose-dependent. The activities of trypsin- and chymotrypsin-like serine proteases were significantly up- and downregulated by high calcium, respectively, in NHEKs. CONCLUSIONS: High calcium, but not 1,25(OH)2 VD3 or RA, increases the expression of serine protease inhibitors in epidermal keratinocytes. Our findings contribute to the understanding of the mechanisms by which serine protease activities are regulated by serine proteases and related inhibitors in epidermal keratinocytes.

Therapeutic Effect of Sirolimus for Lymphangioleiomyomatosis Remaining in the Abdominopelvic Region After Lung Transplantation: A Case Report.

PURPOSE: Sirolimus (SRL) is used to treat pulmonary lymphangioleiomyomatosis (P-LAM). There is limited evidence that SRL has systemic efficacy for the patients with extrapulmonary lymphangioleiomyomatosis (E-LAM) remaining after lung transplantation (LT) for P-LAM. This report examines the efficacy of SRL treatment for the patient with E-LAM remaining after an LT for P-LAM. CASE SUMMARY: The course of the patient's recovery from an LT for P-LAM was complicated by lymphedema in the left femoral region that was caused by two E-LAM lesions remaining in the left pelvic cavity and in the retroperitoneal area. After the LT was performed, the patient started SRL treatment to reduce the E-LAM lesions. The daily SRL dose, selected based on the standard SRL dose for P-LAM, was initiated at 1 mg/d and was maintained at 2 mg/d. The remaining E-LAM lesions and lymphedema in the left femoral region improved in approximately 9 months after the LT with the administration of both SRL and the standard immunosuppressive therapy used by Okayama University Hospital, including tacrolimus, mycophenolate mofetil, and prednisolone. The SRL and tacrolimus trough concentrations in whole blood were maintained within the therapeutic window for the next 1.5 years after initiation of SRL treatment. The patient experienced no severe adverse events that required discontinuation of the SRL treatment during this time. CONCLUSION: The patients with remaining E-LAM lesions may receive SRL treatment to improve the quality of life after LT for P-LAM as effective therapy in cases where the patient's recovery is complicated by E-LAM lesions.

Design study of electron cyclotron resonance-ion plasma accelerator for heavy ion cancer therapy.

Electron Cyclotron Resonance-Ion Plasma Accelerator (ECR-IPAC) device, which theoretically can accelerate multiple charged ions to several hundred MeV with short acceleration length, has been proposed. The acceleration mechanism is based on the combination of two physical principles, plasma electron ion adiabatic ejection (PLEIADE) and Gyromagnetic Autoresonance (GYRAC). In this study, we have designed the proof of principle machine ECR-IPAC device and simulated the electromagnetic field distribution generating in the resonance cavity. ECR-IPAC device consisted of three parts, ECR ion source section, GYRAC section, and PLEIADE section. ECR ion source section and PLEIADE section were designed using several multi-turn solenoid coils and sextupole magnets, and GYRAC section was designed using 10 turns coil. The structure of ECR-IPAC device was the cylindrical shape, and the total length was 1024 mm and the maximum diameter was 580 mm. The magnetic field distribution, which maintains the stable acceleration of plasma, was generated on the acceleration center axis throughout three sections. In addition, the electric field for efficient acceleration of electrons was generated in the resonance cavity by supplying microwave of 2.45 GHz.

SU-E-T-142: Evaluation of the Energy Dependence of Gafchromic EBT3 Film for Electron Beams.

PURPOSE: To evaluate the energy dependence of Gafchromic EBT3 film for establishing a quality assurance method of bolus electron conformal radiotherapy. METHODS: We irradiated electron beam to EBT3 films, which were set in the water tank. The linear accelerator used was Varian Clinac 21EX. The energy of electron beams were 9 and 12 MeV. The irradiated field size was 10×10 cm2 and the source to surface distance was 100 cm. The depths of measurement were 22 (depth of dose maximum; dmax), 31, and 37 mm for 9 MeV and 28 (dmax), 43, and 50 mm for 12 MeV. The irradiated doses were 25, 50, 75, 100, 150, 200, and 300 cGy. EBT3 films were readout with a flat-bed scanner 48 hours after irradiation, and the optical density (OD) curve was obtained for each beam energy and depth. The OD curves were approximated by a third-order polynomial. The doses were evaluated at netOD 0.1 and 0.3 from the approximated curves. RESULTS: The differences of the evaluated doses from those for 9 MeV at 22 mm depth were from 2 to 14 % for netOD=0.1, and from 1 to 13 % for netOD=0.3, respectively. The netOD curves of dmax for both energies showed good agreement, while large discrepancy was found in the deeper depths. CONCLUSIONS: The dependence of dose response of EBT3 film on electron beam energy was small at dmax, while it increased at deeper depth in the present study. It can be considered that the discrepancy was caused by setup error because dose gradient was steeper at the deeper region. In future work, we will perform more precise measurement with a solid phantom to evaluate the energy dependence of EBT3 film.

SU-E-T-497: Study of Restricted Mass Stopping Power Ratio in Small Electron Tube Fields.

PURPOSE: Electron tubes with small radii are useful to treat narrow regions which cannot accommodate normal electron applicators. In small electron fields, it is not trivial to estimate restricted mass stopping power ratio (MSR), which is needed to evaluate dose from ion chamber measurement. We studied MSRs in small electron tube fields using the Monte Carlo simulation. METHODS: Electron tubes with radii, 3 and 2.5 cm, were used in this study. Nominal electron energies were 6 and 9 MeV. There were two types of tubes. One has a normal cut but the other has a 45-degree cut. For the normal cut tube, percent depth dose (PDD) in water was evaluated along the center of axis (CAX) of a beam. For the 45-degree cut tube, PDD was evaluated along the vertical line from the intersection of the CAX and the phantom surface with 45-degree gantry angle. The MSRs and mean electron energies were calculated using the Monte Carlo simulation. RESULTS: We found good agreement between the measured and calculated PDDs. The changes of mean energies from those in the 10×10 cm2 field at the depth of maximum dose (dmax) were very small for the normal cut electron tubes. For the 45-degree cut tubes, the changes of mean energies at dmax were less than 1 MeV. The MSRs in the normal cut tube fields were almost the same as those in the 10×10 cm2 field at the corresponding depths. The MSRs for the 45-degree cut tubes deviated from those in the 10×10 cm2 by about 1% (1.5 % at most). CONCLUSIONS: We evaluated the mean energies and MSRs in small electron tube fields. The deviations of them from the values in the 10×10 cm2 were small. The maximum difference of MSR was 1.5% in 45-degree cut tube fields. This work was supported by KAKENHI (23791449), Japan Society for the Promotion of Science, and Cancer Professional Training Plan, The Ministry of Education, Culture, Sports, Science and Technology, Japan.

Cloning, expression and purification of extracellular serine protease Esp, a biofilm-degrading enzyme, from Staphylococcus epidermidis.

AIMS: Staphylococcus epidermidis Esp, an extracellular serine protease, inhibits Staphylococcus aureus biofilm formation and nasal colonization. To further expand the biotechnological applications of Esp, we developed a highly efficient and economic method for the purification of recombinant Esp based on a Brevibacillus choshinensis expression-secretion system. METHODS AND RESULTS: The esp gene was fused with the N-terminal Sec-dependent signal sequence of the B. choshinensis cell wall protein and a C-terminal hexa-histidine-tag gene. The recombinant Esp was expressed and secreted into the optimized medium as an immature form and subsequently activated by thermolysin. The mature Esp was easily purified by a single purification step using nickel affinity chromatography and showed proteolytic activity as well as Staph. aureus biofilm destruction activity. CONCLUSIONS: The purification yield of the developed extracellular production system was 5 mg recombinant mature Esp per 20-ml culture, which was much higher than that of an intracellular production system in Escherichia coli (3 mg recombinant Esp per 1-l culture). SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings will be a powerful tool for the production and purification of recombinant Esp and also applicable to a large variety of recombinant proteins used for basic researches and biotechnological applications.

CCR2 regulates monocyte recruitment as well as CD4 T1 allorecognition after lung transplantation.

Graft rejection remains a formidable problem contributing to poor outcomes after lung transplantation. Blocking chemokine pathways have yielded promising results in some organ transplant systems. Previous clinical studies have demonstrated upregulation of CCR2 ligands following lung transplantation. Moreover, lung injury is attenuated in CCR2-deficient mice in several inflammatory models. In this study, we examined the role of CCR2 in monocyte recruitment and alloimmune responses in a mouse model of vascularized orthotopic lung transplantation. The CCR2 ligand MCP-1 is upregulated in serum and allografts following lung transplantation. CCR2 is critical for the mobilization of monocytes from the bone marrow into the bloodstream and for the accumulation of CD11c(+) cells within lung allografts. A portion of graft-infiltrating recipient CD11c(+) cells expresses both recipient and donor MHC molecules. Two-photon imaging demonstrates that recipient CD11c(+) cells are associated with recipient T cells within the graft. While recipient CCR2 deficiency does not prevent acute lung rejection and is associated with increased graft infiltration by T cells, it significantly reduces CD4(+) T(h)1 indirect and direct allorecognition. Thus, CCR2 may be a potential target to attenuate alloimmune responses after lung transplantation.

Transvaginal mesh (TVM) reconstruction with TVT/TOT sling for vaginal prolapse concurrent with stress urinary incontinence.

We evaluated clinical efficacies of transvaginal mesh (TVM) reconstruction alone and those concomitant with a TVT/TOT sling for the treatment of pelvic organ prolapse (POP) and stress urinary incontinence (SUI). Between January 2006 and February 2007, 138 female patients with POP underwent TVM reconstruction. The mean age was 66.6 years (range: 52-84). Fourteen individuals were qualified as grade II in the POP quantification (POP-Q) system, 85 and 39 were grades III and IV, respectively. One hundred and seventeen of 138 (85 %) cases showed SUI. Twenty-one patients without SUI underwent TVM alone, and 117 cases with SUI underwent TVM concomitant with TVT/TOT sling. Mean operation time and intra-operative bleeding was 79 min (range: 25-177) and 74 ml (range: 10-429), respectively. Mean follow-up period is 5.3 months (range: 1-14). The vaginal prolapses were cured (grade 0) in 129 cases (93 %) after the surgery. Total inter-national prostate symptom score (IPSS), its QOL score, International Consultation on Incontinence Questionnaires Short Form (ICIQ-SF) significantly improved (from 12.6 to 3.9; p < 0.0001, from 5.0 to 1.0; p < 0.0001, and from 6.1 to 2.5; p < 0.01, respectively). Six of 21 cases (29%) who underwent TVM alone showed postoperative de-novo SUI. In contrast, 116 cases (99%) who underwent TVM concomitant with TVT/TOT, experienced a cure of SUI. Maximum flow rate did not change postoperatively in the both groups. In conclusion, the short-term efficacies of TVM reconstruction for POP are excellent, and a concomitant TVT/TOT sling prevents postoperative SUI.

Costimulatory blockade-mediated lung allograft acceptance is abrogated by overexpression of Bcl-2 in the recipient.

Lung allografts are considered to be more immunogenic than other solid organs. Little is known about the effectiveness of immunosuppressive regimens after lung transplantation. Herein, we describe a novel model of murine vascularized orthotopic lung transplantation we used to study the effects of costimulatory blockade on lung rejection. Transplants were performed in the Balb --> B6 strain combination. Recipients were either not immunosuppressed or received perioperative CD40/CD40L and CD28/B7 costimulatory blockade. Nonimmunosupressed Balb/c --> B6 lung transplants had severe acute rejection 7 days after transplantation and CD8(+) T cells outnumbered CD4(+) T cells within the allografts. Alternatively, B6 recipients that received perioperative costimulatory blockade had minimal inflammation and there were nearly equal numbers of CD8(+) and CD4(+) T cells in these grafts. Approximately one third of graft-infiltrating CD4(+) T cells expressed Foxp3. CD4(+) T cells isolated from these grafts induced apoptosis of alloreactive CD8(+) T cells that were stimulated with donor splenocytes in vitro. In contrast with wild-type B6 recipient mice, we observed severe rejection of Balb/c lungs 7 days after transplantation into Bcl-2 transgenic B6 recipients that had received costimulatory blockade. CD8(+) T cells outnumbered CD4(+) T cells in these immunosuppressed Bcl-2 transgenic recipients and, compared with immunosuppressed wild-type B6 recipients, a lower percentage of graft-infiltrating CD4(+) T cells expressed Foxp3, and a higher percentage of graft-infiltrating CD8(+) T cells expressed intereferon-gamma. Thus, our results show that perioperative blockade of the CD40/CD40L and CD28/B7 costimulatory pathways markedly ameliorates acute rejection of lung allografts in wild type but not Bcl-2 transgenic recipients.

Monocyte differentiation is controlled by MyD88 after mouse orthotopic lung transplantation.

In lung grafts, ischemia-reperfusion signals rapidly induce the recruitment and differentiation of host monocytes into macrophages and dendritic cells. The nature of ischemia-reperfusion signals are antigen independent, but have been hypothesized to initiate Toll-like receptor (TLR) and interleukin (IL)-1R-mediated signaling pathways that are thought to potentiate alloimmune responses. We wondered whether MyD88, an adaptor molecule critical for both TLR and IL-1R-mediated inflammatory responses, regulated monocyte differentiation in a mouse model of vascularized orthotopic lung transplantation. Orthotopic left lung transplants were performed in the following syngeneic combinations: CD45.1(+) B6 --> CD45.2(+) MyD88(-/-) and CD45.1(+) B6 --> CD45.2(+) B6. One day later, recipient-derived dendritic cells and macrophage numbers were assessed in the bronchiolar lavage by FACS analysis. Compared with the bronchiolar lavage of wildtype recipients, MyD88(-/-) recipients had lower numbers of dendritic cells in lung graft airways that were of recipient origin. Lower numbers of newly differentiated lung graft dendritic cells was coincident with the appearance of higher numbers of undifferentiated monocytes in the lung airways of MyD88(-/-) recipients as compared with wild-type recipients. Moreover, adoptive transfer experiments demonstrated that MyD88(-/-) monocytes were poorer at differentiating into lung dendritic cells as compared with wild-type monocytes. Taken together, these data show that MyD88 regulates graft-infiltrating monocyte differentiation and suggests a mechanism by which TLR/IL-1R-signaling pathways control adaptive responses in lung allografts through controlling monocyte fate.

Selective production of cis-9,trans-11 isomer of conjugated linoleic acid from trans-vaccenic acid methyl ester by Delacroixia coronata.

AIMS: Bio-process development for isomer selective and efficient production of cis-9,trans-11-octadecadienoic acid (CLA) from trans-vaccenic acid (t-VA, trans-11-octadecenoic acid) through microbial fatty acid Delta9-desaturation reaction. METHODS AND RESULTS: A total of 550 strains of fungi and yeasts were screened for CLA production from t-VA through Delta9 desaturation. Delacroixia coronata IFO 8586 was selected as a potent producer of CLA from t-VA. Efficient CLA production was observed during cultivation in medium supplemented with the methyl ester of t-VA (t-VAME). Under the optimal conditions with 33.3 mg ml(-1) of t-VAME as substrate, 10.5 mg ml(-1) CLA was produced by D. coronata IFO 8586 after 7 days of cultivation in the medium containing dextrin (5.0%), tryptone (2.0%) and thiourea (0.83 micromol ml(-1)). The strain produced the cis-9,trans-11 isomer of CLA selectively (98% of total CLA), with a small amount of the trans-9,trans-11 isomer (2% of total CLA), mainly in the form of triacylglycerols (69% of total CLA). CONCLUSIONS: A practical bio-process for selective production of cis-9,trans-11 isomer of CLA using filamentous fungus D. coronata IFO 8586 was successfully established. SIGNIFICANCE AND IMPACT OF THE STUDY: Isomer selective bio-process for the practical production of cis-9,trans-11-CLA was first established. The process is benefitable for expanding the application of CLA for medicinal and nutraceutical purposes.

Identification and cornification-related gene expression of canine keratinocyte differentiation-associated protein, Kdap.

The outermost layer of skin, the epidermis, is cornified epithelial tissue composed of keratinocytes. To maintain the structure and function of the epidermis, the regulation of proliferation, differentiation, and cornification of keratinocytes is crucial, and various soluble factors secreted by keratinocytes are involved in these regulations. Previously, work has shown that keratinocytes secreted the protein Kdap (keratinocyte differentiation-associated protein) associated with the formation of cornified cell envelopes, a specialized protective barrier structure on the periphery of terminally differentiating keratinocytes. In the present report, the canine counterpart of human Kdap is identified and an attempt has been made to define its physiological role in canine keratinization. Canine Kdap (cKdap) showed structural features commonly observed in other counterparts and is secreted from transfected cells. The expression profile of cKdap mRNA, which was restrictively expressed in cornified epithelial tissues besides skin has also been determined. These findings indicate that there is a strong association between cKdap expression and cornification, which supports previous observations that Kdap is involved in the synthesis and/or degradation of cornified cell envelopes in humans and mice.

[Sacral neuromodulation in treatment of functional disorders of the lower urinary tract. An overview of basic principles, indications, outcomes]. / Sakrale Neuromodulation in der Behandlung von Funktionsstörungen des unteren Harntrakts. Ein Uberblick über Prinzip, Indikationen und Ergebnisse.

Over the last few years, sacral neuromodulation has become an established treatment option for dysfunctions of the lower urinary tract. It fills the gap that used to exist between conservative therapy and costly invasive methods such as urinary drainage via a deactivated bowel segment. Initially, the clinical value of sacral neuromodulation was controversial even among neurourologists. This was mainly due to a lack of understanding of the physiological processes, uncertain diagnostics, the design of the hardware, and a surgical topography relatively unknown to the urologist. In the meantime, however, sacral neuromodulation has become a standard part of clinical routine with respect to the treatment of dysfunctions of the lower urinary tract, and it is regularly employed in various urological institutions across Europe and the USA. This form of treatment, which is the final straw for patients who believed themselves-after many frustrated therapy attempts-to be "hopeless cases," can now also successfully be employed as an ambulatory measure. The latest data from our hospital, as well as contributions presented at the last DGU Congress in Wiesbaden, indicate that patients with neurogenic urinary retention are the most likely to profit from this treatment option.

Serum stem cell growth factor for monitoring hematopoietic recovery following stem cell transplantation.

Stem cell growth factor (SCGF) is a novel cytokine for primitive hematopoietic progenitor cells. Although it has burst-promoting activity and granulocyte/macrophage colony-promoting activity in vitro, its significance in hematopoiesis in vivo has not been elucidated. In this study, we have established enzyme-linked immunosorbent assay (ELISA) to quantify human SCGF and measured serum cytokines in normal volunteers and 27 patients undergoing stem cell transplantation (SCT), including six autologous and 21 allogeneic transplants. SCGF levels gradually increased after SCT regardless of graft-versus-host disease or type of transplant. The maximum level of SCGF was observed during the rapid granulocyte recovery phase in patients subjected to an autologous transplantation, and during the granulocyte stabilization phase in allogeneic patients. SCGF levels in PBSCT patients began to rise earlier than in BMT patients. Two patients with no increment of SCGF after SCT showed delayed engraftment. The source of SCGF was further analyzed by RT-PCR and we found that SCGF was highly expressed in bone marrow (BM) CD34(+) and CD34(-)CD33(+) cells, but not in BM CD34(-)CD33(-) cells, BM stromal cells and peripheral blood cells. The cell population expressing SCGF in BM possess the colony-forming cell activity. Therefore, serum SCGF can be an indicator of hematopoietic recovery following SCT.

[Treatment of strategy for traumatic disruption of the thoracic aorta accompanied by multiple trauma].

Disruption of the thoracic aorta due to blunt chest trauma is often fatal and should generally be treated surgically as soon as possible. However, cases of disruption of the thoracic aorta due to blunt chest trauma are often complicated by damage to vital organs, making treatment difficult. Our policy is to treat other organs before treating the thoracic aorta in 1) cases in which fracture of the pelvic bone or bleeding from abdominal cavity organs is causing shock and 2) cases of severe cerebral contusion or intracranial hemorrhage that require emergency surgical treatment. The use of a stent for treatment of acute-stage cases should be considered prudently.

[Perioperative management of patient with pulmonary operation associated with interstitial pneumonia].

The prognosis of patients complicated with interstitial pneumonia (IP) who undergo pulmonary operations is very poor if acute exacerbation occurs after the operation. We have experienced 6 cases in which operations were performed on lungs complicated with IP The first patient died due to acute exacerbation, but the subsequent 5 patients had good postoperative courses due to 1) short-term administration of low-dose steroid before and after the operation, 2) avoidance of administration of high concentration of oxygen during and after the operation, and 3) administration of erythromycin before and after the operation, 4) N-acetylcysteine inhalation and administration of dl-alpha-tocopherylnicotinate before and after the operation. It is thought that careful management of such patients is needed to prevent acute exacerbation even after discharge from hospital.