Expression of human cathelicidin peptide LL-37 in inflammatory bowel disease.

Cathelicidin peptide LL-37 plays an important role in the early host response against invading pathogens via its broad-spectrum anti-microbial activity. In this study, we investigated LL-37 expression in the inflamed mucosa of inflammatory bowel disease (IBD) patients. Furthermore, the regulatory mechanism of LL-37 induction was investigated in human colonic subepithelial myofibroblasts (SEMFs). LL-37 mRNA expression and protein secretion were analysed using real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Intracellular signalling pathways were analysed using immunoblotting and specific small interference RNA (siRNA). The expression of LL-37 mRNA was increased significantly in the inflamed mucosa of ulcerative colitis and Crohn's disease. The Toll-like receptor (TLR)-3 ligand, polyinosinic-polycytidylic acid (poly(I:C), induced LL-37 mRNA expression and stimulated LL-37 secretion in colonic SEMFs. The transfection of siRNAs specific for intracellular signalling proteins [Toll/IL-1R domain-containing adaptor-inducing interferon (IFN) (TRIF), tumour necrosis factor receptor-associated factor (TRAF)6, transforming growth factor ß-activated kinase (TAK)1] suppressed the poly(I:C)-induced LL-37 mRNA expression significantly. Poly(I:C)-induced phosphorylation of mitogen-activated protein kinases (MAPKs) and activated nuclear factor kappa B (NF-κB) and activating factor protein (AP)-1. siRNAs specific for NF-κB and c-Jun inhibited poly(I:C)-induced LL-37 mRNA expression. LL-37 suppressed lipopolysaccharide (LPS)-induced interleukin (IL)-6 and IL-8 expression significantly in colonic SEMFs. The expression of LL-37 was up-regulated in the inflamed mucosa of IBD patients. LL-37 was induced by TLR-3 stimulation and exhibited an anti-microbial effect via interaction with lipopolysaccharide (LPS).

Increase in the Magnetic Ordering Temperature (Tc) as a Function of the Applied Pressure for A2Mn[Mn(CN)6] (A = K, Rb, Cs) Prussian Blue Analogues.

Magnetization measurements under pressure reveal that the external hydrostatic pressure significantly increases in the ferrimagnetic transition temperature, Tc, for A2Mn[Mn(CN)6] (A = K, Rb, Cs). In the case of monoclinic A = K and Rb, dTc/dp values are 21.2 and 14.6 K GPa-1, respectively, and Tc increases by 53 and 39%, respectively, from ambient pressure to 1.0 GPa. The cubic A = Cs compound also shows a monotonous increase with an initial rate of 4.22 K GPa-1 and about 11.4 K GPa-1 above 0.6 GPa, and an overall Tc increase by 26% at 1.0 GPa. The increase in Tc is attributed to deformation of the structure such that the MnII-N≡C angle decreases with increasing pressure. The smaller the alkali cation, the greater the decrease in the MnII-N≡C angle induced by pressure and the larger the increase of dTc/dp. This is in accordance with the ambient-pressure structures for A2Mn[Mn(CN)6] (A = K, Rb, Cs), which have decreasing MnII-N≡C angles that correlate to the observed increasing Tcs as K > Rb > Cs. The large increase in Tc for the A = K compound is the highest class among several cyano-bridged metal complexes. The tuning of the transition temperature by such a weak pressure may lead to additional applications such as switching devices.

Tortuosity is the Significant Predictive Factor for Renal Branch Occlusion after Branched Endovascular Aortic Aneurysm Repair.

OBJECTIVE: After multi-branched endovascular aneurysm repair (mbEVAR), renal branch occlusion is the most frequent form of branch failure. Pre-operative renal angulation and post-operative morphology of the renal branch were quantified and their impact on occlusion was analyzed. METHODS: Patients who underwent mbEVAR between January 2010 and December 2013 were reviewed retrospectively. Only renal branches constructed with caudally directed cuffs were included. Patients without post-operative computed (CT) angiography were excluded. The main outcome was the primary patency of the renal branches. The renal angulation and the morphology of renal branch (bridging length, renal coverage length, tortuosity index, and angulation of distal renal artery) were quantified using CT. The impacts of morphology, implanted stents, and patient characteristics were investigated by time to event analyses. RESULTS: Ninety renal arteries in 49 patients were enrolled. Median follow up was 12 months (IQR 6-20 months). Balloon expandable stent grafts were used in 93% (84/90) of renal branches. Self expandable stent grafts were used in 12. Ninety-one percent (82/90) were lined with self expandable bare stents. Ten branches occluded after 8 months (median; IQR 1-14 months). Four of them underwent re-interventions, achieving secondary patency. The median renal angulation was -10° (IQR -40 to 0). The median bridging length was 42 mm (IQR 39-46 mm) and renal coverage 17 mm (IQR 12-22 mm). Median tortuosity index was 1.11 (IQR 1.04-1.19). The angulation of the distal renal artery was 140.7 ± 20.5°. In multivariate analysis, a tortuosity index > 1.11 was identified as the only significant predictor for occlusion (hazard ratio: 4.94; 95% CI: 1.01-24.30, p = .04). CONCLUSIONS: High tortuosity was a significant predictor for the occlusion of renal branches, but renal angulation, bridging length, and the extent of renal coverage were not. By avoiding highly tortuous renal branch paths, good outcomes are expected even in upwardly directed renal arteries. Longer paths are acceptable.

Positioning bedridden patients to reduce interface pressures over the sacrum and great trochanter.

OBJECTIVE: In this study, we evaluated the effect of hip-joint rotation on the interface pressure over the sacrum and greater trochanter with a new protocol for positioning of bedridden elderly patients. METHOD: The interface pressure values over the sacrum and greater trochanter in bedridden patients were evaluated. These were collected in the supine position, 90° lateral position, and 30° and 40° laterally inclined positions with external rotation or neutral positioning of the hip joint. Each interface pressure was assessed with a device measuring pressure distribution, after which, the peak pressure index (PPI) was calculated. RESULTS: In the 17 patients examined, the PPI over the sacrum in the supine position was significantly greater than that in other positions. In the 30° and 40° laterally inclined positions, the PPIs over the greater trochanter were significantly lower in the neutral position of the hip joint compared with those in the external rotation position. CONCLUSION: Our findings revealed the effects of hip-joint rotation on the interface pressure for the greater trochanter, possibly due to the increased distance between the greater trochanter and the sacrum caused by neutral position of the hip joint. The results demonstrate that it is to best place the hip joint in a neutral position when the legs are in contact with the bed in order to distribute the pressure over the greater trochanter in the 30° and 40° laterally inclined positions. These results can be applied to the clinical setting to improve patient positioning and decrease pressure ulcers. DECLARATION OF INTEREST: The authors declare that they have no competing financial interests.

Survival of HER2-positive primary breast cancer patients treated by neoadjuvant chemotherapy plus trastuzumab: a multicenter retrospective observational study (JBCRG-C03 study).

We investigated the disease-free survival (DFS) of HER2-positive primary breast cancer patients treated with neoadjuvant chemotherapy plus trastuzumab, as well as predictive factors for DFS and pathologic response. Data from 829 female patients treated between 2001 and 2010 were collected from 38 institutions in Japan. Predictive factors were evaluated using multivariate analyses. The 3-year DFS rate was 87 % [95 % confidence interval (CI) 85-90]. The pathologic complete response (pCR: ypT0/is + ypN0) rate was 51 %. The pCR rate was higher in the ER/PgR-negative patients than in the ER/PgR-positive patients (64 vs. 36 %, P < 0.001). Patients with pCR showed a higher DFS rate than patients without pCR (93 vs. 82 %, P < 0.001). Multivariate analysis revealed three independent predictors for poorer DFS: advanced nodal stage [hazard ratio (HR) 2.63, 95 % CI 1.36-5.21, P = 0.004 for cN2-3 vs. cN0], histological/nuclear grade 3 (HR 1.81, 95 % CI 1.15-2.91, P = 0.011), and non-pCR (HR 1.98, 95 % CI 1.22-3.24, P = 0.005). In the ER/PgR-negative dataset, non-pCR (HR 2.63, 95 % CI 1.43-4.90, P = 0.002) and clinical tumor stage (HR 2.20, 95 % CI 1.16-4.20, P = 0.017 for cT3-4 vs. cT1-2) were independent predictors for DFS, and in the ER/PgR-positive dataset, histological grade of 3 (HR 3.09, 95 % CI 1.48-6.62, P = 0.003), clinical nodal stage (HR 4.26, 95 % CI 1.53-13.14, P = 0.005 for cN2-3 vs. cN0), and young age (HR 2.40, 95 % CI 1.12-4.94, P = 0.026 for ≤40 vs. >40) were negative predictors for DFS. Strict pCR (ypT0 + ypN0) was an independent predictor for DFS in both the ER/PgR-negative and -positive datasets (HR 2.66, 95 % CI 1.31-5.97, P = 0.006 and HR 3.86, 95 % CI 1.13-24.21, P = 0.029, respectively). These results may help assure a more accurate prognosis and personalized treatment for HER2-positive breast cancer patients.

Clinical outcomes after infrainguinal bypass grafting for critical limb ischaemia in patients with dialysis-dependent end-stage renal failure.

OBJECTIVE: To assess and risk-stratify the medium-term clinical outcomes after infrainguinal bypass grafting (IBG) to treat critical limb ischaemia (CLI) in patients with end-stage renal disease. METHODS: This was a retrospective single-centre study. Between April 2007 and March 2011, 112 limbs from 89 patients were studied. In particular, amputation-free survival (AFS), 30 day mortality, freedom from major adverse limb events (MALE), limb salvage, and overall survival were examined. The aim was to identify outcome predictors. RESULTS: Eight patients (9%) died within 30 days of IBG. The only positive predictor of 30-day mortality was an ejection fraction (EF) < 40% (hazard ratio [HR] 5.57, 95% confidence interval [CI] 1.16-26.83; p = .03). The mean follow-up duration was 14 months. The 1- and 2-year AFS rates were 64% and 43%, respectively, and the rates of freedom from MALE were 81% and 77%, respectively. In addition, the 1- and 2-year limb salvage rates were 89% and 85%, and the survival rates were 68% and 50%, respectively. Non-ambulatory status was negatively associated with AFS (HR 3.04, 95% CI 1.59-5.82; p < .01), freedom from MALE (HR 4.98, 95% CI 1.91-12.96; p < .01), and limb salvage (HR 5.18, 95% CI 1.47-18.30; p = .01). The other negative predictors of overall survival were a serum albumin level <3.0 g/dL (HR 2.26, 95% CI 1.12-4.58; p = .02) and an EF <40% (HR 2.24, 95% CI 1.05-4.79; p = .04). CONCLUSION: Patients with CLI on dialysis enjoyed satisfactory freedom from MALE and limb salvage, but survival and AFS were significantly less than reported for IBG in patients with CLI who did not receive dialysis. In addition, patients with an EF <40%, lower serum albumin (<3.0 g/dL), or non-ambulatory status experienced particularly poor clinical outcomes after IBG.

Astaxanthin ameliorates heat stress-induced impairment of blastocyst development in vitro:--astaxanthin colocalization with and action on mitochondria--.

PURPOSE: The effects of astaxanthin (Ax) on the in vitro development of bovine embryos cultured under heat stress were investigated in combination with the assessment of its cellular accumulation and action on mitochondrial membrane potential (ΔΨm). METHODS: Bovine ≥8-cell embryos were collected on day 3 after in vitro fertilization and exposed to single (day 4) or repeated (day 4 and 5) heat stress (10 h/day at 40.5 °C). Ax was added into culture medium under the repeated heat stress and blastocyst development was evaluated. The cellular uptake of Ax in embryos was examined using bright-field and confocal laser-scanning microscopy, and high-performance liquid chromatography. The relationship between Ax and mitochondria localization was assessed using MitoTracker dye. The effects of Ax on ΔΨm were investigated using JC-1 dye. RESULTS: Blastocyst development in the repeated heat stress treatment decreased significantly (P < 0.05) compared with those in single heat stress or normal thermal treatment. The addition of Ax into culture medium did lead to a significant recovery in blastocyst development in the repeated heat-treated group. Ax was detected in cytoplasm of embryos and observed to colocalize with mitochondria. Ax recovered ΔΨm in embryos that was decreased by the heat treatment. CONCLUSIONS: Ax ameliorated the heat stress-induced impairment of blastocyst development. Our results suggest that the direct action of Ax on mitochondrial activity via cellular uptake is a mechanism of the ameliorating effects.

Predictions of the pathological response to neoadjuvant chemotherapy in patients with primary breast cancer using a data mining technique.

Nomogram, a standard technique that utilizes multiple characteristics to predict efficacy of treatment and likelihood of a specific status of an individual patient, has been used for prediction of response to neoadjuvant chemotherapy (NAC) in breast cancer patients. The aim of this study was to develop a novel computational technique to predict the pathological complete response (pCR) to NAC in primary breast cancer patients. A mathematical model using alternating decision trees, an epigone of decision tree, was developed using 28 clinicopathological variables that were retrospectively collected from patients treated with NAC (n = 150), and validated using an independent dataset from a randomized controlled trial (n = 173). The model selected 15 variables to predict the pCR with yielding area under the receiver operating characteristics curve (AUC) values of 0.766 [95 % confidence interval (CI)], 0.671-0.861, P value < 0.0001) in cross-validation using training dataset and 0.787 (95 % CI 0.716-0.858, P value < 0.0001) in the validation dataset. Among three subtypes of breast cancer, the luminal subgroup showed the best discrimination (AUC = 0.779, 95 % CI 0.641-0.917, P value = 0.0059). The developed model (AUC = 0.805, 95 % CI 0.716-0.894, P value < 0.0001) outperformed multivariate logistic regression (AUC = 0.754, 95 % CI 0.651-0.858, P value = 0.00019) of validation datasets without missing values (n = 127). Several analyses, e.g. bootstrap analysis, revealed that the developed model was insensitive to missing values and also tolerant to distribution bias among the datasets. Our model based on clinicopathological variables showed high predictive ability for pCR. This model might improve the prediction of the response to NAC in primary breast cancer patients.

Effects of folic acid on the development and oxidative stress of mouse embryos exposed to heat stress.

The development of mammalian pre-implantation embryos is inhibited by heat stress, and the inhibitory effect is associated with excess reactive oxygen species (ROS). Folate is a nutrient with various physiological functions including antioxidative effects. We first investigated the transcript expression for 10 enzymes in the cycle of folate metabolism (folate-methionine cycle) in mouse embryos at the 1-cell, 2-cell, 4- to 8-cell, morula and blastocyst stages using reverse transcription-polymerase chain reaction. All of the transcripts were consistently expressed, except for Mat1a, which was not detected from the 4- to 8-cell stage onward. Next, the effects of folic acid (the synthetic form of folate) on the development and ROS levels of heat-stressed embryos were investigated. One-cell mouse embryos were cultured with or without 1000 ng/ml folic acid basically at 38°C, and in the heat-stressed groups, embryos were exposed to 39.5°C/10 h/day on the first two days of culture. The heat stress significantly (p < 0.05) decreased blastocyst development and cell number and increased ROS levels compared to those in the group not subjected to heat stress; however, among the heat-stressed groups, blastocyst development and cell number were increased and the ROS level was decreased by the addition of folic acid. These results indicate that the mRNA of folate-methionine cycle enzymes are expressed in mouse pre-implantation embryos, suggesting they can independently utilize folate, and the inhibitory effects of heat stress on the development of mouse pre-implantation embryos are ameliorated by folic acid. The ameliorating effects of folic acid may be partly due to its antioxidative property.

Optimum microcurrent stimulation intensity for galvanotaxis in human fibroblasts.

OBJECTIVE: In this study, we develop methods to measure galvanotaxis of fibroblasts and determined the optimum conditions of electrical stimulation. METHOD: An inverted 35mm dish containing cell suspensions (3×105 primary human skin fibroblasts, DMEM, and 10% FBS) was placed on the centre of a 100mm dish. The 35mm dish was removed 24 hours later, and culture medium was added to the 100mm dish. Fibroblasts were randomised (double-blind) into three groups, where electrical stimulation was given at varying intensities: 0UA (control), 50UA, and 100UA. Electrical stimulation (frequency=0.3Hz) was conducted, for a duration of 4 hours, with platinum electrodes in a CO2 incubator. We took pictures immediately before and 20 hours after stimulation. We calculated the migration ratio to the negative pole by dividing the area of attached fibroblasts after stimulation with that before stimulation. RESULTS: The migration ratio to the negative pole was significantly higher in the 100UA group than in the control group (p<0.05). The ratios were 0.902±0.292 in the control group, 1.128±0.253 in the 50UA group, and 1.24±0.300 in the 100UA group. CONCLUSION: This study observed the change in cell proliferation during the initial 24-hour period after plating and was thus able to quantitatively evaluate the migration. The results suggest that a low-intensity direct current promotes migration to the negative pole of human dermal fibroblasts, which is charged with positive electricity. Several clinical reports using the methods in this study showed the microcurrent efficacy for pressure ulcer healing. Electrical stimulation based on our in vitro experiment might be important for the development of physical therapy for pressure ulcers.

Enhanced clinical mastitis resistance in Holsteins with a FEZL p.Gly105(12_13) polymorphism.

Mastitis is a common infectious disease of the mammary gland and a major problem in the dairy industry. We previously reported that forebrain embryonic zinc finger-like (FEZL) encoding a stretch of 12 glycines (p.Gly105[12]) instead of 13 glycines (p.Gly105[13]) is associated with a lower somatic cell score (SCS) in a family derived from Walkway Chief Mark. Here we report that the p.Gly105[12] allele is associated with a significantly decreased incidence of clinical mastitis in a large Holstein population. We genotyped the FEZL polymorphism in 918 randomly collected Holstein sires, and investigated the effect of the polymorphism on the estimated breeding value (EBV) for SCS and milk, fat, solids-not-fat, and protein yield, and on the number of cattle with clinical mastitis among daughters derived from these sires. The average EBV for SCS among sires carrying the heterozygous p.Gly105[12] was significantly lower than that among sires carrying the homozygous p.Gly105[13], whereas we found no unfavorable effects of this polymorphism on EBV for milk, fat, solids-not-fat, and protein yield. The proportion of cows with clinical mastitis derived from sires carrying heterozygous p.Gly105[12] was significantly lower than that of daughters derived from sires carrying the homozygous p.Gly105[13]. Thus, selection of sires carrying p.Gly105[12] could be beneficial in the dairy industry by reducing the incidence of mastitis.

Accuracy of Le Fort I osteotomy with combined computer-aided design/computer-aided manufacturing technology and mixed reality.

The aim of this study was to verify the reproducibility and accuracy of preoperative planning in maxilla repositioning surgery performed with the use of computer-aided design/manufacturing technologies and mixed reality surgical navigation, using new registration markers and the HoloLens headset. Eighteen patients with a mean age of 26.0 years were included. Postoperative evaluations were conducted by comparing the preoperative virtual operation three-dimensional image (Tv) with the 1-month postoperative computed tomography image (T1). The three-dimensional surface analysis errors ranged from 79.9% to 97.1%, with an average error of 90.3%. In the point-based analysis, the errors at each point on the XYZ axes were calculated for Tv and T1 in all cases. The median signed value deviation of all calculated points on the XYZ axes was -0.03mm (range -2.93mm to 3.93mm). The median absolute value deviation of all calculated points on the XYZ axes was 0.38mm (range 0mm to 3.93mm). There were no statistically significant differences between any of the points on any of the axes. These values indicate that the method used was able to reproduce the maxilla position with high accuracy.

Glomerular hyperfiltration with hyperglycemia in the spontaneously diabetic Torii (SDT) fatty rat, an obese type 2 diabetic model.

Glomerular hyperfiltration is observed in an early stage of kidney diseases including diabetic nephropathy. A better understanding of pathophysiological changes in glomerular hyperfiltration is essential for development of new therapies to prevent kidney disease progression. In this study, we investigated glomerular changes including glomerular filtration rate (GFR) and glomerular size in the Spontaneously Diabetic Torii (SDT) fatty rat, an obese type 2 diabetic model, and we also evaluated pharmacological effects of the sodium glucose cotransporter 2 inhibitor dapagliflozin on the renal lesions. Dapagliflozin was administered to SDT fatty rats from 5 to 17 weeks of age. Blood and urinary biochemical parameters were periodically measured. GFR was determined by transdermal GFR monitor at 16 weeks of age and histopathological analysis was performed at 17 weeks of age. SDT fatty rat developed severe hyperglycemia and exhibited pathophysiological abnormalities in the kidney, such as an increased GFR, glomerular hypertrophy and tissue lesions. Dapagliflozin achieved good glycemic control during the experimental period, inhibited the increase in GFR, and improved histopathological abnormalities in tubules. These results suggest that the SDT fatty rat is a useful model for analyzing the pathogenesis of diabetic nephropathy during its early stage and dapagliflozin improves not only hyperglycemia but also glomerular hyperfiltration and tubule lesions in SDT fatty rat.

Transforming growth factor beta induces IgA production and acts additively with interleukin 5 for IgA production.

Effects of transforming growth factor beta (TGF-beta) on IgA production by LPS-stimulated B cells have been studied. TGF-beta itself could augment polyclonal IgA production in concomitant inhibition of polyclonal IgM and IgG1 production. Furthermore, TGF-beta and IL-5 additively augmented IgA production. TGF-beta exerted its activity early in the culture (by 2 d in a 5-d culture) and IL-5 was required late in the culture. Surface IgA- (sIgA-) B cells responded to TGF-beta for the development of IgA-secreting cells. By contrast, sIgA+ B cells, but not sIgA- B cells, responded to IL-5 for IgA production. These results suggest that TGF-beta has a differential role in the induction of IgA production from IL-5 on murine-activated B cells.

Effects of astaxanthin-containing oil on development and stress-related gene expression of bovine embryos exposed to heat stress.

Early bovine embryos are vulnerable to heat stress during the first few days after fertilization. The inhibitory effect of heat stress on embryonic development is known to be associated with oxidative stress, which can be attenuated by antioxidants. In the present study, we focused on the use of astaxanthin as an antioxidant and examined the effects of astaxanthin-containing oil (Ax) on post-fertilization development of bovine embryos subjected to heat stress in vitro and the expression of stress-related genes. Bovine 1-cell embryos were in vitro produced by in vitro maturation and fertilization (IVF) of oocytes recovered from abattoir-derived ovaries. At 20 h post-insemination (hpi, 0 h = the start of IVF), the embryos were introduced in modified synthetic oviduct fluid supplemented with 25 ppm of Ax (concentration of astaxanthin was 0.25 ppm) or vehicle (dimethyl sulfoxide) up to 72 hpi. The embryos were basically cultured at 38.5°C, and in the heat stress group, embryos were exposed twice to 40.5°C for 10 h (at 20-30 and 44-54 hpi). Under the condition without the Ax treatment, the cleavage rate, rate of development to the 5-8 cell stage, blastocyst yield from cultured embryos and that from cleaved embryos were lower in the heat stress group than in the group not subjected to heat stress (p < 0.05). In the heat stress group, the rate of development to the 5-8 cell stage was improved (p < 0.05) by the addition of Ax. Subsequently, we performed semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) to investigate the effects of heat stress and Ax on the mRNA expression of Src homology 2 domain-containing transforming protein C1 (SHC1), an oxidative stress adaptor protein, and superoxide dismutase 2 (SOD2), a mitochondrial reactive oxygen species (ROS) scavenger. In 5-8 cell embryos at 72 hpi, the mRNA expression levels of SHC1 and SOD2 were lower in the Ax- and heat-treated group than in the other groups (p < 0.05). These results suggest that Ax added to the culture medium ameliorates the embryonic development impaired by heat stress with its altering effects on the expression of stress-related genes.

Dynamic expression of vimentin in bovine blastocysts in extended in vitro culture.

In mammalian blastocysts, the inner cell mass (ICM) differentiates into the epiblast and the hypoblast. In vitro culture of bovine pre-implantation embryos is generally carried out for a limited duration at most up to the blastocyst formation, therefore, it is unclear whether and when the differentiation of ICM occurs in vitro. In bovine species, epiblast formation is characterized by the expression of the intermediate filament protein vimentin. In the present study, the spatial and temporal expression of vimentin in bovine blastocysts maintained under extended in vitro culture (EIVC) was investigated. Bovine blastocysts produced by in vitro maturation, fertilization, and culture were further cultured in vitro in modified synthetic oviduct fluid up to day 12 (fertilization = day 0). Early, expanded, or hatched blastocysts on day 8 and hatched blastocysts on days 9 and 12 were individually subjected to indirect immunofluorescence analysis with an anti-vimentin monoclonal antibody and to semi-quantitative reverse transcription-polymerase chain reaction analysis to examine the vimentin expression. Most of the blastocysts (25/26) did not exhibit any significant immunostaining for the vimentin protein on day 8. On the other hand, the vimentin protein was observed in 59.1% (13/22) and 86.4% (19/22) of the hatched blastocysts on days 9 and 12, respectively. Vimentin was detected as filaments localized within a portion of the ICM, and its expression levels varied among the embryos. Vimentin transcript could not be detected in 50% (3/6) of the blastocysts on day 8, whereas it was detected in all the blastocysts on days 9 and 12. Examination of the day 8 blastocysts revealed that compared with the slow developers, the fast developers (blastocysts which had already expanded by 168 h post-insemination) had significantly higher levels of vimentin transcripts. These results indicate that the ICM differentiates dynamically in bovine blastocysts maintained under EIVC, as reflected by the expression of vimentin and that the higher vimentin expression may reflect the higher developmental competence of embryos.

Evaluation of the combined use of ultrasound irradiation and wound dressing on pressure ulcers.

OBJECTIVE: To evaluate the effect of ultrasound irradiation when used alongside standard care in the treatment of pressure ulcers; outcome measures were reduction in wound size and exudate weight. METHOD: Five patients (two male and three female, age range: 76-92 years) with seven ulcers participated in this study. They had National Pressure Ulcer Advisory Panel (NPUAP) stage III or IV pressure ulcers. We conducted an ABABA study (A: standard treatment with dressings that promote a moist wound healing environment; B: ultrasound irradiation administered to the pressure ulcer through the same dressing used in period A; each period lasted 2-4 weeks). Six ulcers each were randomised to either the treatment group or control group. One ulcer was not randomised, but was the first to receive ultrasound in the BABA sequence, with a view to determining if the pilot was feasible. The control group received sham ultrasound in period B. Pulsed ultrasound (20% duty cycle, 0.5W/cm2 on the wound surface, 1MHz or 3MHz, for 10 minutes) was applied five times weekly. RESULTS: In the treatment group, two ulcers markedly decreased in size after 3-4 weeks of US treatment, one ulcer decreased in size soon after initiation of treatment and one ulcer showed no clear reduction in size. The volume of exudate was greater in period B than A in two ulcers that reduced markedly in size after 3-4 weeks of US treatment. None of the ulcers in the control group decreased markedly in size. CONCLUSION: This pilot study suggests that US used alongside standard treatment might promote the healing of pressure ulcers. However, larger studies are required to determine the efficacy and mechanism of US treatment for PUs. DECLARATION OF INTEREST: None.

[Stress response genes expression analysis of barley Hordeum vulgare under space flight environment].

Transcriptome of barley Hordeum vulgare grown aboard International Space Station (ISS) was analyzed by means of microarray. It was revealed 500 genes with mRNA level, changed more than two folds in space environment. Among them are genes encoding stress response proteins, videlicet Heat Shock Proteins (HSP), Pathogenesis-Related Proteins (PR) and Antioxidant Proteins. Further analysis of these genes by real time PCR showed enhanced transcription level of Reactive oxygen Species (ROS) scavenging genes. The mRNA level of superoxide dismutase (sod) was 6 folds higher in space environment when compare to Earth conditions. Glutamyl transferase gene expression was enhanced 24 times in space. Transcription of catalase gene (cat) was increased 18 times and of ascorbate peroxidase was increased 3 times in space in comparison with ground control. For the first time it was shown that space flight environment may induce oxidative stress in plants.

Commissioning of high current H+/D+ ion beams for the prototype accelerator of the International Fusion Materials Irradiation Facility.

The Linear IFMIF (International Fusion Materials Irradiation Facility) Prototype Accelerator (LIPAc) is aiming at demonstrating the low energy section of a 40 MeV/125 mA IFMIF deuteron accelerator up to 9 MeV with a full beam current in cw operation. For such a high-power beam, the LIPAc injector is required to produce a 100 keV D+ beam with 140 mA and match it for injection into the Radio Frequency Quadrupole (RFQ) accelerator. The injector is designed by CEA-Saclay based on the high intensity light ion source (SILHI). In 2019, the commissioning of the RFQ to demonstrate the D+ beam acceleration at a low duty cycle (0.1%) was conducted. A nominal beam current of 125 mA D+ beam was accelerated up to 5 MeV through the RFQ successfully. The LIPAc injector fully satisfied the requirements for RFQ beam commissioning at the pulse mode.