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The present study examined the associations among physical activity during different times of the day(i.e., morning, midday,evening) and days of the week(i.e., weekdays, weekend days) with restless legs syndrome(RLS) severity and sleep quality in adults with multiple sclerosis(MS).Participants(N = 39) completed the International Restless Legs Syndrome Study Group Scale and the Pittsburgh Sleep Quality Index as measures of RLS severity and global sleep quality, respectively, and wore an ActiGraph GT3X + on the non-dominant hip for seven days for measuring physical activity(i.e., light physical activity[LPA], moderate-to-vigorous physical activity[MVPA], and steps).There was a significant effect of time of day wherein participants had more midday activity compared with morning or evening for LPA, MVPA, and steps; there was no effect of day of the week. Spearman's rho correlations across the week indicated higher evening LPA, evening steps, and overall daily steps (ρ = -0.42,ρ = -0.51,ρ = -0.40,respectively) were associated with lower RLS severity. Similar associations were demonstrated for weekdays(ρ = -0.39,ρ = -0.50,ρ = -0.41,respectively) and weekend days(ρ = -0.44,ρ = -0.51,ρ = -0.39,respectively). Higher evening MVPA on weekend days(ρ = -0.42) was associated with lower RLS severity. Higher morning and overall daily LPA(ρ = 0.34,ρ = 0.33,respecitvely) were associated with worse sleep quality. Similar associations were demonstrated on weekdays(ρ = 0.33,ρ = 0.37,respectively) and more evening MVPA(ρ = 0.32) was associated with worse sleep quality. On weekend days, more morning MVPA(ρ = 0.42) was associated with worse sleep quality.Our results suggest that undertaking physical activity, particularly LPA and steps, in evenings may be important for managing symptoms of RLS without worsening sleep quality.

To examine device-measured physical activity levels and sedentary behavior participation during different times of the day (ie, morning, midday, and evening) in adults with multiple sclerosis (MS) who differed in fatigue status.Cross-sectional survey.Remote survey study managed by a university-based research laboratory.A population-based sample of 1000 participants with MS were sent recruitment materials by the North American Research Committee on Multiple Sclerosis and 218 participants completed all relevant outcomes (N=218).Not applicable.Participants (N=218) completed the Fatigue Severity Scale (FSS) as a measure of fatigue severity and were divided into subgroups of fatigued (FSS score ≥4) and nonfatigued (FSS score4). Participants wore an ActiGraph GT3X+ on the nondominant hip for 7 days to measure physical activity (ie, light physical activity [LPA], moderate-to-vigorous physical activity [MVPA], steps) and sedentary behavior.Fatigued participants engaged in less MVPA (F[1216]=18.5, P.001), fewer steps (F[1216]=27.8, P.001), and more sedentary time (F[1216]=8.2, P=.005) than nonfatigued participants. Regardless of group, the highest levels of LPA (F[1.7355.7]=72.9, P.001) and MVPA (F[1.8395.3]=23.0,P.001) occurred in the morning and middle of the day, with the lowest levels in the evening. Regardless of group, the highest levels of sedentary behavior occurred in the evening, with similar levels in the morning and evening (F[1.6354.5]=84.3,P.001). Regardless of group, participants had more steps in the middle of the day, followed by morning and then evening (F[1.8383.9]=84.7,P.001).Our results suggest that physical activity timing should be considered in future development and delivery of behavior interventions that focus on increasing physical activity and reducing sedentary behavior among adults with MS who have fatigue.

Background: Nipple-sparing mastectomy (NSM) is an increasingly popular surgical procedure for treatment of breast cancer. However, NSM is controversial because of its association with locoregional recurrence. We started performing NSM in 1978. Methods: We investigated the surgical safety including nipple necrosis, and nipple–areola recurrence (NAR) and skin flap recurrence (SFR) after NSM for 1071 patients with breast cancer, including 31 with stage 0, 414 with stage 1, 479 with stage 2, 141 with stage 3 and 6 with stage 4, from 1985 to 2017. Our NSM method involved creating a thick skin flap to avoid surgical complications. No patients received radiotherapy. In 1034 patients with stage 1–3 breast cancer treated with NSM who developed NAR or SFR, we evaluated cancer stage, nuclear grade, lymph node metastasis, tumor–nipple–areola distance, and histological classification as tubule forming, solid and scirrhous type. In 748 patients with early stage 1 and 2A breast cancer treated with NSM, NAR and SFR were evaluated for estrogen receptor and HER2 expression. We evaluated disease-free interval and frequency of late NAR and SFR. Results: Median follow-up after NSM was 87 (3–397) months. There was only one case of total nipple necrosis among all 1071 patients. There were 96 patients (9.0%) with local recurrence, including 44 (4.0%) with NAR and 52 (4.8%) with SFR. NAR was seen in 1 (3.1%), 14 (3.4%), 17 (5.1%), 5 (3.4%), 7 (5.0%) and 0 patients with stage 0, 1, 2A, 2B, 3 and 4 cancer, respectively. SFR was seen in 0, 15 (3.6%), 8 (2.7%), 7 (4.8%), 22 (15.6%) and 0 patients with stage 0, 1, 2A, 2B, 3 and 4 cancer, respectively. Median disease-free interval of NAR and SFR was 3.4 (0.96–22.3) and 2.5 (0.21–21.2) years, respectively. Twenty-three (53%), 12 (27.9%) and 6 (14%) patients had NAR at more than 3, 5 and 10 years after NSM, respectively. Twenty (38%), 13 (25%) and 6 (11.5%) patients had SFR at more than 3, 5 and 10 years after NSM, respectively. Therefore, late NAR and SFR were observed. Patients with stage 1–3 cancer treated with NSM who had significantly more frequent NAR, were characterized by high nuclear grade and tubule-forming type cancer. Patients with significantly more frequent SFR were characterized by stage 3 cancer, positive lymph node metastasis and age ≤40 years. Patients with early stage breast cancer treated with NSM with significantly more frequent NAR had negative estrogen receptor expression, positive HER2 expression and shorter tumor–nipple–areola distance (≤2 cm). Overall survival was significantly better in patients with NAR (97% at 5 years and 80% at 10 years) than SFR (71% at 5 years and 50% at 10 years). Regarding SFR, overall survival was significantly worse for multiple (≥2) and diffuse (clinical inflammatory syndrome) recurrence than for single-nodule recurrence. There was no significant difference in prognosis between NAR and single-nodule SFR. Conclusions: Our data showed that clinicopathological features and prognosis differed between patients with NAR and SFR. There was no significant difference in prognosis between NAR and single-nodule SFR. Late NAR and SFR were seen, and careful long-term follow-up observation is necessary after NSM. Citation Format: Sakurai T, Suzuma T, Yoshimura G, Sasaki E, Umemura T, Sakurai T. Clinicopathological features and prognosis of nipple–areola and skin flap recurrence after nipple-sparing mastectomy for breast cancer over 20 years [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-14-08.

Defining off-target profiles of gene-editing nucleases and CRISPR base editors remains an important challenge for use of these technologies, therapeutic or otherwise. Existing methods can identify off-target sites induced by these gene editors on an individual genome but are not designed to account for the broad diversity of genomic sequence variation that exists within populations of humans or other organisms. Here we describe OligoNucleotide Enrichment and sequencing (ONE-seq), a novel in vitro method that leverages customizable, high-throughput DNA synthesis technology instead of purified genomic DNA (gDNA) from individual genomes to profile gene editor off-target sites. We show that ONE-seq matches or exceeds the sensitivity of existing single-genome methods for identifying bona fide CRISPR-Cas9 off-target sites in cultured human cells and in vivo in a liver-humanized mouse model. In addition, ONE-seq outperforms existing best-in-class single-genome methods for defining off-target sites of CRISPR-Cas12a nucleases, cytosine base editors (CBEs), and adenine base editors (ABEs), unveiling previously undescribed bona fide off-target sites for all these editors in human cells. Most importantly, we leveraged ONE-seq to generate the first experimentally-derived population-scale off-target profiles for Cas9 nucleases that define the impacts of sequence variants from >2,500 individual human genome sequences in the 1000 Genomes Project database. Notably, some of the variants we identified that lead to increased mutation frequencies at off-target sites are enriched in specific human populations. We validated that novel population-specific, variant-sensitive off-target sites nominated by ONE-seq in vitro can show increased frequencies of mutations in human lymphoblastoid cells (LCLs) harboring these sequence variants. Collectively, our results demonstrate that ONE-seq is a highly sensitive off-target nomination method that can uniquely be used to identify population subgroup-linked differences in off-target profiles of gene editors. ONE-seq provides an important new pathway by which to assess the impacts of global human genetic sequence diversity on the specificities of gene editors, thereby enabling a broader and more all-inclusive approach for profiling off-target effects of these transformative therapeutic technologies.

BACKGROUND Wearable motion sensors are gaining popularity for monitoring free-living physical activity among people with Parkinson disease (PD), but more evidence supporting the accuracy and precision of motion sensors for capturing step counts is required in people with PD. OBJECTIVE This study aimed to examine the accuracy and precision of 3 common consumer-grade motion sensors for measuring actual steps taken during prolonged periods of overground and treadmill walking in people with PD. METHODS A total of 31 ambulatory participants with PD underwent 6-min bouts of overground and treadmill walking at a comfortable speed. Participants wore 3 devices (Garmin Vivosmart 3, Fitbit One, and Fitbit Charge 2 HR), and a single researcher manually counted the actual steps taken. Accuracy and precision were based on absolute and relative metrics, including intraclass correlation coefficients (ICCs) and Bland-Altman plots. RESULTS Participants walked 628 steps over ground based on manual counting, and Garmin Vivosmart, Fitbit One, and Fitbit Charge 2 HR devices had absolute (relative) error values of 6 (6/628, 1.0%), 8 (8/628, 1.3%), and 30 (30/628, 4.8%) steps, respectively. ICC values demonstrated excellent agreement between manually counted steps and steps counted by both Garmin Vivosmart (0.97) and Fitbit One (0.98) but poor agreement for Fitbit Charge 2 HR (0.47). The absolute (relative) precision values for Garmin Vivosmart, Fitbit One, and Fitbit Charge 2 HR were 11.1 (11.1/625, 1.8%), 14.7 (14.7/620, 2.4%), and 74.4 (74.4/598, 12.4%) steps, respectively. ICC confidence intervals demonstrated low variability for Garmin Vivosmart (0.96 to 0.99) and Fitbit One (0.93 to 0.99) but high variability for Fitbit Charge 2 HR (–0.57 to 0.74). The Fitbit One device maintained high accuracy and precision values for treadmill walking, but both Garmin Vivosmart and Fitbit Charge 2 HR (the wrist-worn devices) had worse accuracy and precision for treadmill walking. CONCLUSIONS The waist-worn sensor (Fitbit One) was accurate and precise in measuring steps with overground and treadmill walking. The wrist-worn sensors were accurate and precise only during overground walking. Similar research should inform the application of these devices in clinical research and practice involving patients with PD.

A quench detection method of a high-temperature superconducting (HTS) coil using a cowinding method is studied. It is experimentally shown using a small YBCO test coil that the cowinding method is highly effective in canceling the inductive voltage component and can detect a quench occurring in an HTS coil with high sensitivity, compared with a commonly used bridge-balance method. An HTS coil is damaged by a quench when hot-spot temperature exceeds a certain threshold. The authors also experimentally investigated conditions to keep the hot-spot temperature of coils wound of YBCO tapes below a safe level, using the highly sensitive cowinding quench detection method.

This paper presents the shear lag phenomenon occurred in steel box moment connections by considering the local effect of column flange flexibility. The stress concentration due to shear lag in steel box member is one of the major concerns for steel box moment connections. When welding is utilized, the connections are susceptible to cracking or failure in the weld parts. Therefore, the maximum stress in the steel box moment connection is always checked during a preliminary design stage. In this study, the stresses due to shear lag were evaluated using least-work solution, considering the flexibility of column flange. The local flexibility of the connection, which caused by the column flange and diaphragms, is represented by an axial spring model, and contributed to stress distribution in the beam flange. Using the presumed longitudinal displacement functions for both the beam flange and web, the stress concentrations were evaluated to increase significantly compared with that obtained from a cantilever beam model. The finite element assessment was also conducted to check the validity of the manual prediction of the stress in various ranges of the beam and column section properties. The results are further summarized, discussed, and showed that more flexible column flange and diaphragm provided higher stress concentrations.

Background The development of CRISPR genome editing has transformed biomedical research. Most applications reported thus far rely upon the Cas9 protein from Streptococcus pyogenes SF370 (SpyCas9). With many RNA guides, wildtype SpyCas9 can induce significant levels of unintended mutations at near-cognate sites, necessitating substantial efforts toward the development of strategies to minimize off-target activity. Although the genome-editing potential of thousands of other Cas9 orthologs remains largely untapped, it is not known how many will require similarly extensive engineering to achieve single-site accuracy within large genomes. In addition to its off-targeting propensity, SpyCas9 is encoded by a relatively large open reading frame, limiting its utility in applications that require size-restricted delivery strategies such as adeno-associated virus vectors. In contrast, some genome-editing-validated Cas9 orthologs are considerably smaller and therefore better suited for viral delivery. Results Here we show that wildtype NmeCas9, when programmed with guide sequences of the natural length of 24 nucleotides, exhibits a nearly complete absence of unintended editing in human cells, even when targeting sites that are prone to off-target activity with wildtype SpyCas9. We also validate at least six variant protospacer adjacent motifs (PAMs), in addition to the preferred consensus PAM (5′-N4GATT-3′), for NmeCas9 genome editing in human cells. Conclusions Our results show that NmeCas9 is a naturally high-fidelity genome-editing enzyme and suggest that additional Cas9 orthologs may prove to exhibit similarly high accuracy, even without extensive engineering. Electronic supplementary material The online version of this article (10.1186/s13059-018-1591-1) contains supplementary material, which is available to authorized users.

This paper presents a Finite Element Analysis(FEA)-based design of a health monitoring system for a structurally complex long-span bridge. The system consists of Bridge Weigh-in-Motion(BWIM), damage detection function and thermal behavior monitoring. To determinate the sensor position for BWIM, the strain response in the floor system is analyzed. Besides, FEA simulations are performed to grasp the damageable parts during an earthquake. Based on the analysis, a sensor arrangement for the bridge is proposed.

Background The family of Janus kinases (JAKs) plays important roles in signalling pathway mediated by various cytokine receptors. An aberrant activation of JAK-STAT signalling has been reported to be involved in the pathogenesis of autoimmune diseases1. Pan-JAK inhibitors have shown a good efficacy in patient with rheumatoid arthritis (RA)2. However, their use is limited due to safety concerns, including severe herpes zoster infection, by inhibiting JAK1-mediated interferon signaling3. Therefore, a selective JAK3 inhibitor would provide a better balance between efficacy and safety than pan-JAK inhibitors. Objectives We identified the characteristics of TAS8274, a novel highly selective inhibitor of JAK3, using in vitro assays, a mouse model of collagen-induced arthritis (CIA), and a mouse model of herpes simplex virus (HSV)−1 infection. Methods In vitro biochemical assay was performed using available kinase assay panels. The effects on anti-inflammatory responses were assessed by examining cytokine productions. IL-2, IL-3, and IFN-α–induced phosphorylation of STAT proteins in peripheral blood mononuclear cells (PBMCs) were analysed by a flow cytometry method. NK cell cytotoxicity in the presence of IFN-α was evaluated by Cr51 release assay. In a mouse skin HSV-1 infection model, TAS8274 and tacrolimus were administered for 7 days before inoculation of the virus on the back skin, and then were administered for another ten consecutive days. At the end of this experiment, the number of papules on the back was counted. To evaluate the therapeutic efficacy using mouse CIA model, TAS8274 was orally administered to CIA mice after the disease onset. Disease severity was evaluated by clinical score of paw swelling, and the scores of inflammation, pannus, cartilage, and bone damage were performed using a modified Mankin score system. Results TAS8274 inhibited the enzymatic activity of JAK3 (IC50=0.16 nM), and showed more than 1000-fold selectivity against other JAK kinases. In the cell-based assays, TAS8274 strongly inhibited IL-17 production from differentiated Th17 cells. TAS8274 also suppressed the IL-2-induced STAT5 phosphorylation in PBMCs, but had much lower inhibitory effects on the IFN-α-induced STAT1 phosphorylation. In contrast, Tofacitinib and Baricitinib had robust inhibitory effects on the IFN-α-induced STAT1 phosphorylation. Furthermore, Tofacitinib and Baricitinib dose-dependently reduced the NK cell cytotoxicity, while TAS8274 had little effect on that. Tacrolimus-treated group significantly increased the number of papules compared with vehicle-treated group in a mouse HSV-1 infection model, but TAS8274-treated group did not increase the number of papules. In an established mouse CIA model, TAS8274 dose-dependently reduced the severity of arthritis and histopathological scores compared with vehicle-treated mice. Conclusions TAS8274 did not inhibit the JAK3-independent STAT signalling pathway in vitro and showed potent efficacy at dose range without exacerbation of the risk of HSV-1 infection. Our study demonstrates that TAS8274 would be an attractive therapeutic agent with excellent balance between efficacy and safety. References [1] Immunity2012;36,542–50. [2] Ann Rheum Dis. 2013;72,111–5. [3] Arthritis Rheumatol2014;66:2675–84. Disclosure of Interest None declared

The most probable cause of quenches of a coil wound of Bi/Ag-sheathed wires is the appearance of a long resistive zone caused by the temperature rise of the long part of the conductor due to, for example, ac losses and the malfunction of the cryogenic system. Protection characteristics of quenches by this cause are experimentally and analytically investigated. In the experiment, the operating temperature of a coil wound of a Bi/Ag-sheathed wire is gradually increased until the resistive voltage of the coil exceeds a given threshold to start a quench dump sequence. Damages in the test coils are examined by changing the decay time constant of the coil current. Based on those results, conditions to protect coils from damages are investigated. Analysis is conducted to simulate quench events, and conditions for quench protection are obtained. The analytical results well explain the quench behaviors of the test coils.

BackgroundThe development of CRISPR genome editing has transformed biomedical research. Most applications reported thus far rely upon the Cas9 protein from Streptococcus pyogenes SF370 (SpyCas9). With many RNA guides, wild-type SpyCas9 can induce significant levels of unintended mutations at near-cognate sites, necessitating substantial efforts toward the development of strategies to minimize off-target activity. Although the genome-editing potential of thousands of other Cas9 orthologs remains largely untapped, it is not known how many will require similarly extensive engineering to achieve single-site accuracy within large (e.g. mammalian) genomes. In addition to its off-targeting propensity, SpyCas9 is encoded by a relatively large (~4.2 kb) open reading frame, limiting its utility in applications that require size-restricted delivery strategies such as adeno-associated virus vectors. In contrast, some genome-editing-validated Cas9 orthologs (e.g. from Staphylococcus aureus, Campylobacter jejuni, Geobacillus stearothermophilus and Neisseria meningitidis) are considerably smaller and therefore better suited for viral delivery.ResultsHere we show that wild-type NmeCas9, when programmed with guide sequences of natural length (24 nucleotides), exhibits a nearly complete absence of unintended editing in human cells, even when targeting sites that are prone to off-target activity with wildtype SpyCas9. We also validate at least six variant protospacer adjacent motifs (PAMs), in addition to the preferred consensus PAM (5’-N4GATT-3’), for NmeCas9 genome editing in human cells.ConclusionsOur results show that NmeCas9 is a naturally high-fidelity genome editing enzyme and suggest that additional Cas9 orthologs may prove to exhibit similarly high accuracy, even without extensive engineering.

Background The erosions of bone and cartilage are a cardinal feature of rheumatoid arthritis (RA) and associated with disease severity and poor functional outcome1. Although several anti-inflammatory drugs improve symptoms of articular inflammation, they are less effective against bone erosion. The bone erosions in RA are associated with aberrant activations of osteoclasts induced by pro-inflammatory cytokines and receptor activator of nuclear factor κB ligand (RANKL)2. Bruton9s tyrosine kinase (BTK), which is expressed in immune cells and mature osteoclast, is reported to be a key molecule in inflammatory response and bone resorption3,4. Thus, targeting BTK may be efficacious against not only inflammation but also bone erosion through direct regulation of activation of effector cells such as B cells, macrophages and osteoclasts in RA. Objectives In this study, we evaluated the effect of TAS5315, a novel BTK inhibitor, on in vitro osteoclasts activation and bone erosion in mouse collagen-induced arthritis. Methods Kinase selectivity of TAS5315 was assessed by available kinase assay panels. The effects of TAS5315 on macrophages and osteoclasts were assessed by examining phosphorylation of BTK, cytokine productions, osteoclast differentiation and bone resorptions. The effects of TAS5315 were investigated in mouse collagen-induced arthritis (CIA). Disease severity was evaluated by clinical score of paw swelling. Changes in bone mineral density (BMD) and bone erosion were assessed using microCT. TNF blocker was used as a control drug. Results TAS5315 selectively inhibited the enzyme activity of BTK and had less off target inhibition against other kinases. TAS5315 dose-dependently inhibited cytokine productions by macrophages, phosphorylation of BTK, osteoclastogenesis and bone resorbing activity in osteoclasts. In established mouse CIA, TAS5315 significantly ameliorated paw swelling in a dose dependent manner and the anti-inflammatory effect of TAS5315 (0.3 mg/kg, once daily) was comparable to that of TNF blocker. Most importantly, improvement of BMD and bone erosion were observed in TAS5315 treated mice at a doses of higher than 0.1 mg/kg within 13 days from treatment initiation, but not in TNF blocker-treated mice. The onset of action of TAS5315 on BMD and bone erosion was earlier and stronger compared with that of TNF blocker. These data suggest that TAS5315 had direct effect against osteoclasts function and led to improvement of bone erosion in murine model for RA. Conclusions Our study demonstrates that TAS5315, a novel BTK inhibitor, would be an ideal RA therapeutic agent that could inhibit bone destruction as well as inflammation. References Nat Rev Rheumatol. 2012;8,656–64. Nat Rev Rheumatol. 2015;11,189–94. Cell. 2008;132,794–806. Drug Discov Today. 2014;19,1200–4. Disclosure of Interest None declared

This work investigates current transport characteristics of soldered lap joints of YBCO coated conductors with Cu stabilizers subjected to repeated uni-axial tensile stresses in the longitudinal direction. Data of influence of repeated stresses on transport characteristics clearly show fatigue effect causing damages in YBCO layer. From experimental data, it is considered that the damages occur in edge areas of lap joint sections where stresses are concentrated.

According to visual inspection on the bridge site, the target bridge has major cracks in RC deck in the center span. This certain type of damage could affect structural performance and should be monitored. The structural health monitoring system was set up on the target bridge and the strain-based evaluation could be used to quantitatively determine bridge condition. The FE model of the bridge was modeled with various types of RC cracking pattern to represent the severity of bridge damage. The relationship between damage level and strain measurement is generated, As a result, Bridge monitoring curve based on strain index has been established.

Introduction: Cosmetic outcome is one of the most important goals of surgical treatment for breast cancer. Therefore, to maximize the cosmetic outcome for patients who require a mastectomy, the actions that can be taken from an oncologically safe perspective must be considered. Recent reports have suggested that nipple-sparing mastectomy (NSM) is oncologically as safe as a standard mastectomy and provides a better cosmetic outcome. However, little is known about the outcome of NSM for patients with multifocal breast cancer. Aim: To determine the oncological and technical outcomes of NSM for patients with multifocal breast cancer compared with NSM for patients with unifocal breast cancer using retrospective long-term follow-up data. Patients and Methods: We retrospectively analyzed 799 patients with primary breast cancer who underwent NSM, including 45 patients with multifocal breast cancer and 754 patients with unifocal breast cancer. The indications for NSM in this study were defined as any stage, any tumor size and any tumor–areola distance. Briefly, NSM was performed when no involvement of the nipple areola complex was revealed by radiographic and clinical examinations. Our surgical notes for NSM comprised a thick skin flap method for conserving the breast volume using subcutaneous fat and avoiding surgical complications including nipple necrosis. No patients received radiotherapy. We compared the local recurrence rate (LRR), disease-free survival (DFS), breast cancer-specific survival (BCSS) and overall survival (OS) between the 45 multifocal breast cancer and 754 unifocal breast cancer cases. Results: The numbers of multifocal breast cancer cases in stages 0, 1, 2A, 2B, 3 and 4 were 1, 17, 12, 8, 7 and 0, respectively. The corresponding numbers of unifocal breast cancer cases were 18, 288, 223, 117, 102 and 6, respectively. The median follow-up periods of the multifocal and unifocal breast cancer cases were 108 and 98 months, respectively. No nipple necrosis was recorded in either group (0/45, 0/754). Regarding NSM for patients with multifocal breast cancer, there were 8 patients (17.7%) with local recurrence, including 4 (8.9%) at the nipple and 4 (8.9%) at the skin flap. In NSM for patients with unifocal breast cancer, there were 63 patients (8.3%) with local recurrence, including 26 (3.4%) at the nipple and 37 (4.9%) at the skin flap. There were significant differences between NSM for patients with multifocal and unifocal breast cancer in the LRR (17.7% vs. 8.3%, p = 0.03) and DFS (10 yr, 72.4% vs. 83.8%, p = 0.04), but no significant differences in the BCSS (10 yr, 83.6% vs. 90.9%, p = 0.18) and OS (10 yr, 83.6% vs. 89.5%, p = 0.32). Conclusions: The present results suggest that NSM for patients with multifocal breast cancer can be technically performed as safely as NSM for patients with unifocal breast cancer. Our long-term follow-up data show that survival after NSM for multifocal breast cancer is equal to that after NSM for unifocal breast cancer. However, careful local observation after NSM is required. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-18-08.

We describe the development of neurological signs in four juvenile black-and-white ruffed lemurs (Varecia variegate), housed at a petting zoo in Japan. The clinical course was severe, with three lemurs dying within 1 day of the appearance of clinical signs. The other lemur was treated and survived. Pathological analyses demonstrated meningitis and the presence of gram-negative bacilli in the cerebrum, cerebellum, palatine tonsil and liver. Klebsiella pneumoniae was isolated from the brain of all of the dead lemurs. Multilocus sequence typing analysis showed that all the isolates were sequence type 86 (ST86). To our knowledge, this is the first determination of K. pneumoniae infection in ruffed lemurs of this genus. K. pneumoniae infection may represent a risk to lemurs and people who come into contact with infected animals.

Malignant melanoma of the vagina is a very rare neoplasm and resistant to conventional radiotherapy. We report a case of vaginal malignant melanoma that was locally well controlled by carbon ion radiotherapy. A 55-year-old postmenopausal woman presented with abnormal vaginal bleeding. On pelvic and imaging examinations, an irregular mass of the posterior vaginal wall sized 7.5 × 5 × 5 cm, an enlarged right inguinal lymph node, and two lung metastases were observed. Histologic diagnosis based on positive immunostaining for HMB-45 was malignant melanoma. She received dacarbazine-based chemotherapy and carbon ion radiotherapy for vaginal and inguinal tumor sites with 57.6 Gy equivalent dose per 16 fractions using five ports. Six months later, she was also given carbon ion radiotherapy for regrowing lung metastasis with 52.8 Gy equivalent dose per four fractions using four ports. She died 19 months after initial treatment due to brain metastases. The primary irradiated tumor disappeared completely 12 months after initial treatment. The vaginal tumor, right inguinal lymph node, and lung tumor treated with carbon ion radiotherapy did not show any evidence of recurrence until her death. Carbon ion radiotherapy may be of value for vaginal malignant melanoma as a conservative approach.

Summary Background No adequate second-line chemotherapy regimen for advanced gastric cancer is available. Aims To determine the safety and optimal dose of paclitaxel/irinotecan as a second-line chemotherapy for patients with advanced and recurrent gastric cancer. Patients and methods Sixteen patients with refractory and advanced measurable gastric cancer who were resistant to 5-FU plus cis-diamminedichloroplatinum (CDDP) therapy (FP) were enrolled. Paclitaxel/irinotecan was given intravenously on days 1, 8, and 15 in repeated 4-week cycles. Paclitaxel/irinotecan doses were escalated in a stepwise fashion as follows: 50/40 mg/m2, 50/50 mg/m2, 50/60 mg/m2, 60/60 mg/m2, 60/70 mg/m2 in levels I, II, III, IV and V, respectively. Results Because of one patient with Grade 3 febrile neutropenia at level I, three more patients were enrolled in level I. Doses were consequently escalated, and in level IV, Grade 3 febrile neutropenia occurred in one patient. Since an additional patient in level IV had grade 4 neutropenia, Level IV was judged as the maximum tolerated dose (MTD). The recommended dose and schedule for phase II study is paclitaxel 50 mg/m2 and irinotecan 60 mg/m2 on days 1, 8, and 15 every 4 weeks. Partial response was observed in 4 of 16 patients. Conclusion Paclitaxel/irinotecan combination regimen at the level III dosage was safe and well tolerated.

AIMS This study aimed to identify the main active component of Lactobacillus brevis KB290 (KB290) that is responsible for enhanced cell-mediated cytotoxic activity of mouse splenocytes Live KB290, a probiotic strain derived from a Japanese traditional pickle, was previously reported to modulate innate immune responses as affecting on cell-mediated cytotoxic activity of mouse splenocytes. METHODS AND RESULTS We used live KB290, heat-killed KB290, a derivative strain (Lact. brevis KB392) with different amounts of cell-bound exopolysaccharide (EPS-b), and a crude extract of EPS-b from KB290 cell surface. Female BALB/c mice were fed a diet containing 10(10) CFU live KB290, 10(10) CFU live KB392, 15 mg heat-killed KB290 or 600 μg crude extract of EPS-b for 1 day. Live KB290 (P < 0.01), heat-killed KB290 (P < 0.05) and crude EPS-b at 600 μg (P < 0.05) per mouse significantly enhanced cytotoxic activity; however, live KB392 had no effect. CONCLUSIONS Both live and heat-killed KB290 and crude EPS-b significantly enhanced cytotoxic activity of mouse splenocytes. SIGNIFICANCE AND IMPACT OF THE STUDY We demonstrated that EPS-b produced by KB290 has a critical role in enhancing cell-mediated cytotoxic activity in mouse spleen.

Eddy Current (EC) inspection of corrosion defects in steel elements partially covered by concrete is modeled by 3D FEM analysis. A differential probe model is proposed for increasing sensibility of the inspection when direct contact to the inspected surface is not possible. Defect response and defect detectability for various defect sizes and concrete cover thickness is analyzed with the proposed EC probe. A parametric study was conducted to identify suitable EC probe parameters.

Chicken blastodermal cells (CBC) from Stage X embryos, which were isolated from newly laid, fertile, unincubated eggs, are pluripotent cells and can produce somatic and germline chimeras when injected into recipient stage X embryos. The CBC retain their pluripotential ability for up to 7 d in vitro. The molecular mechanisms that control proliferation and differentiation of CBC are largely unknown, although protein tyrosine kinases (PTK) are known to play important roles in these processes in similar cells. To understand better the molecular mechanisms of proliferation and differentiation in CBC, expression profiles of PTK and stem cell factor (SCF) were analyzed by reverse transcription polymerase chain reaction (RT-PCR) using gene-specific and degenerate oligonucleotide primers. Seventeen distinct PTK, including 14 receptor-type and 3 nonreceptor-type PTK and SCF were identified by RT-PCR. Expression of all of the genes was confirmed by northern blot analysis. The northern blot analysis showed that all probes hybridized with one or more transcripts at various expression levels. The expression of the 17 PTK and SCF genes in CBC suggests that they might play a role in signal transduction pathways that control the proliferation or differentiation in CBC.

Guest Editor's Introduction: This paper was originally presented at the 9th congress of ASAIO in April 1963. The original paper was printed in Volume IX of Trans. Soc. Artif. Int. Organs 1963, pages 358–362. It was reprinted with permission in Therapeutic Apheresis, Volume 4, Issue 1, 2000 as a millennium project of the International Society for Apheresis. This paper describes for the first time a hybrid artificial live and extracorporeal circulatory system composed of a specially developed high permeable gel type cellophane membrane. This membrane separated the patient's blood and the metabolic fluid in which freeze-dried canine liver granules were contained. Carbohydrate metabolism and ammonia detoxification were maintained in this metabolic chamber, eventually helping the patient's metabolic functions. In addition to experimental studies, four clinical studies were briefly reported.

We investigated the effects of the application of wastewater sludges on microbial degradation of crude oil. the experiments were carried out in four beach simulating tanks. In each tank, filled with gravel and seawater, crude oil was added on the surface of seawater. Dehydrated-wastewater sludge, liquid-wastewater sludge and synthetic fertilizers were added in tanks A, D and C, respectively. In tanks A and D, sharp increases in the bacterial number and the oxygen consumption occurred immediately while the increases in tank C was slower. The concentration of nitrogen was high during first few weeks and later decreased in tanks A and D. The synthetic nitrogen fertilizer was maintaining the nitrogen concentration above 1 mg l−1 for the whole experimental period. These observations indicated that the effect of the synthetic nitrogen fertilizer was more enduring than those of the liquid and dehydrated sludges. However, higher concentrations of phosphorous were supplied by both the liquid and dehydrated sludges...

The purposes of this study were monitoring and regulating latissimus dorsi muscle (LDM) performance with muscle internal pressure and thickness, and with burst stimulation parameters, respectively. The unconditioned LDM flap of a pig was connected to a measurement system to examine force and contraction length. Internal pressure was measured using a catheter pressure transducer with a fluid-filled balloon inserted into the muscle, and correlated linearly with force during both contraction and relaxation. During contraction, thickness change correlated linearly with contraction length and the area enclosed within an internal pressure-thickness curve, and the X-axis was correlated linearly with muscle work. Stroke work increased incrementally according to the pulse width up to 6.72 msec or burst rate up to 120 Hz, and then plateaued at 240 Hz. The authors conclude that muscle work during contraction could be monitored with internal pressure and thickness, and that stroke work could be regulated with pulse width or burst rate.

Summary Embryonic stem cells (ESCs) have become increasingly attractive for cell replacement therapies of osteodegenerative diseases; however, pre-clinical studies in large animal models to repair diseased or injured bone are lacking. As a first step into this direction, we describe here the feeder-free cultivation and directed osteogenic differentiation of marmoset ESCs. Introduction Owing to their potential to self-renew and their enormous differentiation capability, ESCs are an adequate cell source for cell replacement therapies. To implement stem cell technology clinically, standardized cultivation and differentiation protocols and appropriate animal models are needed. Here, we describe the feeder-free cultivation of Callithrix jacchus ESCs (cESCs) in a chemically defined medium and their subsequent osteogenic differentiation. Methods cESCs were maintained on mouse embryonic fibroblast feeder layers or in feeder-free conditions with activin A and basic fibroblast growth factor. Differentiation into mature osteoblasts was steered with ascorbic acid, v-glycerophosphate and 1a,25-(OH)2 vitamin D3 employing various induction strategies. Results In feeder-free conditions, cESCs maintained pluripotency as indicated by Oct-4 and Nanog expression, positive immunostaining for typical primate ESC markers and high telomerase activity. Cells also remained karyotypically normal after 40 passages without feeder cells. The hanging drop protocol as well as omitting the embryoid body step proved unsuccessful to initiate osteogenic differentiation. The highest degree of osteogenesis was achieved by formation of embryoid bodies employing the cell cluster technique as indicated by the amount of deposited calcium and bone marker gene expression. Early addition of retinoic acid further improved the yield of osteoblasts and led to an increase in calcium deposition. Conclusions The osteogenic differentiation potential of feeder-free cESCs was equal if not higher compared to cells grown on feeders. These findings open the field for near clinical transplantation studies in primate models to evaluate the effectiveness of ESC-derived osteoblasts.

The research studies applicability of an Eddy Current NDT inspection method for detection of corrosion in steel structural members including at locations where direct access to the steel element surface is not possible. The method is evaluated by 3D numerical analysis. Parametric study is conducted for selection of appropriate inspection probe parameters. The inspection is applied to inspection of corrosion in areas where H-shaped steel truss elements are passing through RC deck.

Background The STOP-Bang questionnaire is used to screen patients for obstructive sleep apnoea (OSA). We evaluated the association between STOP-Bang scores and the probability of OSA. Methods After Institutional Review Board approval, patients who visited the preoperative clinics for a scheduled inpatient surgery were approached for informed consent. Patients answered STOP questionnaire and underwent either laboratory or portable polysomnography (PSG). PSG recordings were scored manually. The BMI, age, neck circumference, and gender (Bang) were documented. Over 4 yr, 6369 patients were approached and 1312 (20.6%) consented. Of them, 930 completed PSG, and 746 patients with complete data on PSG and STOP-Bang questionnaire were included for data analysis. Results The median age of 746 patients was 60 yr, 49% males, BMI 30 kg m−2, and neck circumference 39 cm. OSA was present in 68.4% with 29.9% mild, 20.5% moderate, and 18.0% severe OSA. For a STOP-Bang score of 5, the odds ratio (OR) for moderate/severe and severe OSA was 4.8 and 10.4, respectively. For STOP-Bang 6, the OR for moderate/severe and severe OSA was 6.3 and 11.6, respectively. For STOP-Bang 7 and 8, the OR for moderate/severe and severe OSA was 6.9 and 14.9, respectively. The predicted probabilities for moderate/severe OSA increased from 0.36 to 0.60 as the STOP-Bang score increased from 3 to 7 and 8. Conclusions In the surgical population, a STOP-Bang score of 5–8 identified patients with high probability of moderate/severe OSA. The STOP-Bang score can help the healthcare team to stratify patients for unrecognized OSA, practice perioperative precautions, or triage patients for diagnosis and treatment.

We sought to determine the role of human papillomavirus (HPV) E6 and E7 oncogenes in nuclear β-catenin accumulation, a hallmark of activated canonical Wnt signaling pathway. We used HPV16-positive oropharyngeal cancer cell lines 147T and 090, HPV-negative cell line 040T, and cervical cell lines SiHa (bearing integrated HPV16) and HeLa (bearing integrated HPV18) to measure the cytoplasmic and nuclear β-catenin levels and the β-catenin/Tcf transcriptional activity before and after E6/E7 gene silencing. Repression of HPV E6 and E7 genes induced a substantial reduction in nuclear β-catenin levels. Luciferase assay showed that transcriptional activation of Tcf promoter by β-catenin was lower after silencing. The protein levels of β-catenin are tightly regulated by the ubiquitin/proteasome system. We therefore performed expression analysis of regulators of β-catenin degradation and nuclear transport and showed that seven in absentia homologue (Siah-1) mRNA and protein levels were substantially upregulated after E6/E7 repression. Siah-1 protein promotes the degradation of β-catenin through the ubiquitin/proteasome system. To determine whether Siah-1 is important for the proteasomal degradation of β-catenin in HPV16-positive oropharyngeal cancer cells, we introduced a Siah-1 expression vector into 147T and 090 cells and found substantial reduction of endogenous β-catenin in these cells. Thus, E6 and E7 are involved in β-catenin nuclear accumulation and activation of Wnt signaling in HPV-induced cancers. In addition, we show the significance of the endogenous Siah-1-dependent ubiquitin/ proteasome pathway for β-catenin degradation and its regulation by E6/E7 viral oncoproteins in HPV16-positive oropharyngeal cancer cells. ©2010 AACR.