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OBJECTIVE Cervical nerve 5 palsy can occur following surgery for cervical spine pathology. The prognosis of C5 palsy is generally favorable, and most patients recover useful function. However, some patients do not recover useful strength. Nerve transfers are a potential effective treatment of postoperative severe C5 palsy. This study aimed to further delineate the natural history of recovery from postoperative C5 palsy, determine whether lack of recovery at specific time points predicts poor recovery prognosis, and thereby determine a reasonable time point for referral to a complex peripheral nerve specialist. METHODS The authors conducted a retrospective review of 72 patients who underwent surgery for cervical spondylosis and stenosis complicated by C5 palsy. Medical Research Council (MRC) motor strength grades were recorded preoperatively; immediately postoperatively; at discharge; and at 2 weeks, 3 months, 6 months, and 12 months postoperatively. Univariate and multivariate logistic regression models were used to identify demographic and clinical risk factors associated with recovery of useful strength after severe C5 palsy. RESULTS The mean patient age was 62.5 years, and 36.1% of patients were female. Thirty patients (41.7%) experienced severe C5 palsy with less than antigravity strength (MRC grade 2 or less) at discharge. Twenty-one (70%) of these patients recovered useful strength (MRC grade 3 or greater) at 12 months postoperatively, and 9 patients (30%) did not recover useful strength at 12 months. Of those patients with persistent severe C5 palsy at 3 months postoperatively, 50% recovered useful strength at 12 months. Of those patients with persistent severe C5 palsy at 6 months postoperatively, 25% recovered useful strength at 12 months. No patient with MRC grade 0 or 1 strength at 6 months postoperatively recovered useful strength. A history of diabetes was associated with the occurrence of severe C5 palsy. On multivariate analysis, female sex was associated with recovery of useful strength. CONCLUSIONS Most patients with severe C5 palsy recover useful strength in their C5 myotome within 12 months of onset. However, at 3 months postoperatively, patients with persistent severe C5 palsy had only a 50% chance of recovering useful strength by 12 months. Lack of recovery of useful strength at 3 months postoperatively is a reasonable time point for referral to a complex peripheral nerve center to establish care and to determine candidacy for nerve transfer surgery if severe C5 palsy persists.

Higher doses of olaparib are sufficient to inhibit PAR formation in BRCA1-mutant SUM-149 cells. At higher concentrations of olaparib (1μM), combination treatment of PARP1 inhibition and radiation decreases PAR in a comparable manner to lower doses of olaparib (20nM) in SUM-149 cells.

Extended methods for immunohistochemistry. Antibodies used in immunohistochemistry experiments are listed with each of the corresponding dilutions, retrieval techniques, and positive controls.

Sustained locoregional control of disease is a significant issue in patients with inflammatory breast cancer (IBC), with local control rates of 80% or less at 5 years. Given the unsatisfactory outcomes for these patients, there is a clear need for intensification of local therapy, including radiation. Inhibition of the DNA repair protein PARP1 has had little efficacy as a single agent in breast cancer outside of studies restricted to patients with BRCA mutations; however, PARP1 inhibition (PARPi) may lead to the radiosensitization of aggressive tumor types. Thus, this study investigates inhibition of PARP1 as a novel and promising radiosensitization strategy in IBC. In multiple existing IBC models (SUM-149, SUM-190, MDA-IBC-3), PARPi (AZD2281-olaparib and ABT-888-veliparib) had limited single-agent efficacy (IC50 > 10 μmol/L) in proliferation assays. Despite limited single-agent efficacy, submicromolar concentrations of AZD2281 in combination with RT led to significant radiosensitization (rER 1.12–1.76). This effect was partially dependent on BRCA1 mutational status. Radiosensitization was due, at least in part, to delayed resolution of double strand DNA breaks as measured by multiple assays. Using a SUM-190 xenograft model in vivo, the combination of PARPi and RT significantly delays tumor doubling and tripling times compared with PARPi or RT alone with limited toxicity. This study demonstrates that PARPi improves the effectiveness of radiotherapy in IBC models and provides the preclinical rationale for the opening phase II randomized trial of RT ± PARPi in women with IBC (SWOG 1706, NCT03598257).

This Athena case describes a young girl with a history of a mildly tender flesh-coloured papule on her parietal scalp with surrounding poliosis.

Sustained locoregional control of disease is a significant issue in patients with inflammatory breast cancer (IBC), with local control rates of 80% or less at 5 years. Given the unsatisfactory outcomes for these patients, there is a clear need for intensification of local therapy, including radiation. Inhibition of the DNA repair protein PARP1 has had little efficacy as a single agent in breast cancer outside of studies restricted to patients with BRCA mutations; however, PARP1 inhibition (PARPi) may lead to the radiosensitization of aggressive tumor types. Thus, this study investigates inhibition of PARP1 as a novel and promising radiosensitization strategy in IBC. In multiple existing IBC models (SUM-149, SUM-190, MDA-IBC-3), PARPi (AZD2281-olaparib and ABT-888-veliparib) had limited single-agent efficacy (IC50 > 10 μmol/L) in proliferation assays. Despite limited single-agent efficacy, submicromolar concentrations of AZD2281 in combination with RT led to significant radiosensitization (rER 1.12–1.76). This effect was partially dependent on BRCA1 mutational status. Radiosensitization was due, at least in part, to delayed resolution of double strand DNA breaks as measured by multiple assays. Using a SUM-190 xenograft model in vivo, the combination of PARPi and RT significantly delays tumor doubling and tripling times compared with PARPi or RT alone with limited toxicity. This study demonstrates that PARPi improves the effectiveness of radiotherapy in IBC models and provides the preclinical rationale for the opening phase II randomized trial of RT ± PARPi in women with IBC (SWOG 1706, NCT03598257).

Increased rates of locoregional recurrence are observed in patients with basal-like breast cancer (BC) despite the use of radiation therapy (RT); therefore, approaches that result in radiosensitization of basal-like BC are critically needed. Using patients’ tumor gene expression data from 4 independent data sets, we correlated gene expression with recurrence to find genes significantly correlated with early recurrence after RT. The highest-ranked gene, TTK, was most highly expressed in basal-like BC across multiple data sets. Inhibition of TTK by both genetic and pharmacologic methods enhanced radiosensitivity in multiple basal-like cell lines. Radiosensitivity was mediated, at least in part, through persistent DNA damage after treatment with TTK inhibition and RT. Inhibition of TTK impaired homologous recombination (HR) and repair efficiency, but not nonhomologous end-joining, and decreased the formation of Rad51 foci. Reintroduction of wild-type TTK rescued both radioresistance and HR repair efficiency after TTK knockdown; however, reintroduction of kinase-dead TTK did not. In vivo, TTK inhibition combined with RT led to a significant decrease in tumor growth in both heterotopic and orthotopic, including patient-derived xenograft, BC models. These data support the rationale for clinical development of TTK inhibition as a radiosensitizing strategy for patients with basal-like BC, and efforts toward this end are currently underway.

Purpose Unmet clinical needs in breast cancer (BC) management include the identification of patients at high risk of local failure despite adjuvant radiation and an understanding of the biology of these recurrences. We previously reported a radiation response signature and here extend those studies to identify a signature predictive of recurrence timing (before or after 3 years). Methods and Materials Two independent patient cohorts were used. The training cohort included 119 patients with in-breast tumor recurrence (343 total), and the validation testing cohort had 16 patients with recurrences (112 total). All patients received radiation treatment after breast-conserving surgery. Initial feature selection used Spearman rank correlation, and a linear model was trained and locked before testing and validation. Cox regression was used for univariate and multivariable analyses (UVA and MVA, respectively). Biologically related concepts were identified using gene set enrichment analysis. Results Spearman correlation identified 485 genes whose expression was significantly associated with recurrence time (early vs late). Feature reduction further refined the list to 41 genes retained within the signature. In training, the correlation of score to recurrence time was 0.85 (P value Conclusions We report a breast cancer gene signature that may identify patients unlikely to respond to adjuvant radiation and may be used to predict timing of recurrences with implications for potential treatment intensification and duration of follow-up for women with breast cancer treated with radiation.

The present study preliminarily examined the differences in maximum handling size, prey size and species selectivity of growth hormone transgenic and non-transgenic common carp Cyprinus carpio when foraging on four gastropods species (Bellamya aeruginosa, Radix auricularia, Parafossarulus sinensis and Alocinma longicornis) under laboratory conditions. In the maximum handling size trial, five fish from each age group (1-year-old and 2-year-old) and each genotype (transgenic and non-transgenic) of common carp were individually allowed to feed on B. aeruginosa with wide shell height range. The results showed that maximum handling size increased linearly with fish length, and there was no significant difference in maximum handling size between the two genotypes. In the size selection trial, three pairs of 2-year-old transgenic and non-transgenic carp were individually allowed to feed on three size groups of B. aeruginosa. The results show that the two genotypes of C. carpio favored the small-sized group over the large-sized group. In the species selection trial, three pairs of 2-year-old transgenic and non-transgenic carp were individually allowed to feed on thin-shelled B. aeruginosa and thick-shelled R. auricularia, and five pairs of 2-year-old transgenic and non-transgenic carp were individually allowed to feed on two gastropods species (P. sinensis and A. longicornis) with similar size and shell strength. The results showed that both genotypes preferred thin-shelled Radix auricularia rather than thick-shelled B. aeruginosa, but there were no significant difference in selectivity between the two genotypes when fed on P. sinensis and A. longicornis. The present study indicates that transgenic and non-transgenic C. carpio show similar selectivity of predation on the size- and species-limited gastropods. While this information may be useful for assessing the environmental risk of transgenic carp, it does not necessarily demonstrate that transgenic common carp might have lesser environmental impacts than non-transgenic carp.

Background: Increased rates of locoregional recurrence (LR) have been observed in TNBC despite chemotherapy and radiation (RT). A novel radiosensitizer screen nominated the AR as a promising target for treatment of radioresistant breast cancer, including TNBC. We assessed the activity of seviteronel (Sevi), a selective CYP17 lyase and AR inhibitor in Phase 2 clinical development for advanced breast and prostate cancer, as a potential radiosensitizer in AR+ TNBC model. Methods: Clonogenic survival assays were used to determine the intrinsic RT sensitivity of 21 breast cancer cell (BCC) lines. IC50 values were determined for 130 clinically available compounds and correlation coefficients were calculated using IC50 values and SF-2Gy. Gene expression was measured using RNA Seq or qRT-PCR and protein expression was measured using RPPA arrays. AR function was assessed using functional inhibition with Sevi in MDA-MB-453, ACC-422, ACC-460, SUM-185 (all four AR+ TNBC), MDA-MB-231 (AR- TNBC), and T47D (AR- ER+) BCC lines. Double-stranded DNA (dsDNA) break repair was assessed with γH2AX foci counting. In vivo tumor growth was measured with varying control and treatment groups (16-20 tumors/group). Kaplan-Meier analysis was performed to estimate local control. A Cox proportional hazards model and multi-variate analysis (MVA) were used to determine variables associated with LRF survival. Results: Our novel radiosensitizer screen identified the activity of anti-androgen therapy as a potentially effective strategy for radiosensitization in RT-resistant BCC lines (R2 =0.46, p-value < 0.01) (Speers et al, J Clin Oncol 35, 2017 (suppl; abstr e12102). Heterogeneity in AR expression was identified in human BCC lines and TNBC samples from patients (N=2098). There was a strong correlation between AR RNA expression and protein expression across all BC intrinsic subtypes. AR inhibition using Sevi induced radiation sensitivity in vitro with an enhancement ratio (ER) of 1.24-1.69 in four different AR+ TNBC lines. No such radiosensitization was seen in AR(-) TNBC or ER+, AR(-) BCC lines. Radiosensitization was at least partially dependent on impaired dsDNA break repair with significant delays in dsDNA break repair at 16 and 24 hours in all AR+ TNBC lines examined (p-value < 0.01). AR inhibition with Sevi significantly radiosensitized AR+ TNBC xenografts in mouse models and markedly delayed tumor-volume tripling time (TTT) and tumor growth (MDA-MB-453: median TTT 16.1 days for RT alone vs. not reached after 45 days for Sevi+RT, p-value Conclusions: Our results implicate the AR as a mediator of radioresistance in breast cancer and support the rationale for developing Sevi as a novel radiosensitizing agent in AR+ TNBC. Citation Format: Speers CW, Chandler B, Olsen E, Wilder-Romans K, Moubadder L, Nyati S, Rae J, Hayes DF, Spratt DE, Wahl DR, Eisner J, Feng FY, Pierce LJ. Radiosensitization of androgen receptor (AR)-positive triple-negative breast cancer (TNBC) cells using seviteronel (INO-464), a selective CYP17 lyase and AR inhibitor [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P1-09-05.

This research compares three new methods and measures for mapping and evaluating the transportation segment of a value chain. The three methods are total energy, energy-time and energy-distance. The methods combine practices used in traditional physical distribution system analysis and extended value stream mapping (VSM) analysis. This research concludes that the energy-distance method and measure is the most effective for analysing, comparing,and identifying opportunities for value chain improvement. It also identifies a ‘U-shape’ that is characteristic of the energy transfer pattern during a transportation segment that suggests that damage from transportation may need to be screened for quality at a subsequent value chain node.

Background: Basal-like breast cancer (BC) has the highest rates of local recurrence despite the use of radiation therapy. Therefore, approaches for radiosensitization are critically needed for patients with this subtype of BC. Methods: Four independent datasets were used to correlate gene expression with local recurrence (LR) and Kaplan-Meier analysis validated the impact of TTK expression on LR. The METABRIC dataset was used to determine TTK expression in BC subtypes. Clonogenic survival assays were used to determine the radiosensitization of cell lines after TTK inhibition (TTKi). Mouse models were used to assess TTKi in combination with RT in vivo. DNA damage was quantified using γH2AX staining. HR and NHEJ efficiency assays were performed using HR/NHEJ specific reporter systems. HR competency was also assessed using Rad51 foci formation assays. Rescue experiments were performed using wild-type (WT) and kinase-dead (KD) TTK plasmids in combination with siRNA targeting the UTR region of TTK. Results: Ten genes were found to significantly correlate with early LR (≤3 years) after surgery and radiation across 4 independent datasets (N=896 pts), with TTK, a cell cycle kinase, ranked the highest. Kaplan-Meier survival analysis in multiple cohorts demonstrated that higher than median TTK expression correlates with decrease LR free survival after RT (HR 1.70-2.42, p35 days], p Conclusion: These data support TTKi as a radiosensitizing strategy for clinical development in basal-like BC patients and that radiosensitization is mediated, at least in part, through impaired HR repair. Citation Format: Benjamin Chandler, Leah Moubadder, Cassandra Ritter, Meilan Liu, Meleah Cameron, Kari Wilder-Romans, Amanda Zhang, Andrea Pesch, Anna Michmerhuizen, Nicole Hirsh, Marlie Androsiglio, Tanner Ward, Eric Olsen, Yashar Niknafs, Sofia Merajver, Dafydd Thomas, Powel Brown, Theodore Lawrence, Shyam Nyati, Lori Pierce, Arul Chinnaiyan, Corey Speers. TTK inhibition radiosensitizes basal-like breast cancer through impaired homologous recombination [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6273.

Purpose – The purpose of this paper is to develop insights about the psychological factors that contribute to lean’s success as a holistic and adaptive system. Design/methodology/approach – The authors use the best-selling book “Thinking, Fast and Slow” (TFAS) (2011) by Nobel-prize winner Daniel Kahneman to provide a familiar lens for readers who might not otherwise be familiar with the psychology theories that are used in this paper to study lean conceptually. With this approach, the paper sheds light on psychological factors that tie together many of the philosophies, principles and practices of lean. Findings – The paper shows how lean’s philosophies, principles and practices provide a synergistic and self-reinforcing system that drives employee thinking and actions. TFAS characterizes thought processes as “fast System 1 thinking” that relies on intuition and “slow System 2 thinking” that is more rational and logical. Lean psychology eliminates waste and adds customer value by supporting, enhancing and taking advantage of beneficial fast thinking and motivating and imposing appropriate slow thinking. Originality/value – The authors develop the concept of lean psychology to describe the relationship between psychology theories and lean. By applying lean psychology, organizations can go beyond superficially adopting a checklist of tools and techniques to more fully take advantage of lean and improve their operations performance.

This article shows how an approach, such as A3, can be adapted to describe and analyze the evolution of a strategic plan and its deployment process within a university setting. We used a case to illustrate how strategy deployment would work. The deployment was accompanied by a process, including workshops, surveys, and participation, under the direction of a strategic steering committee. The article begins by building the context for the strategic process from a historical perspective for the university in the study. A change management model was employed to supplement and evaluate the A3 approach. The research findings indicate that the A3 approach was a useful tool in the beginning stages of a strategic deployment process; however, as more stakeholders become involved, other organizational change dynamics came into play and needed to be addressed.

A structurally transformed lytic bacteriophage having a broad host range of Staphylococcus aureus strains and a penicillin-binding protein (PBP 2a) antibody conjugated latex beads have been used to produce a biosensor for discrimination of methicillin resistant (MRSA) and sensitive (MSSA) S. aureus strains. The lytic phages have been converted into phage spheroids and moved onto a biosensor surface by Langmuir-Blodgett technique. The produced biosensors have been tested by a quartz crystal microbalance with dissipation tracking to examine bacteria-phage interactions. Bacteria-spheroid interactions caused reduction of resonance frequency and rise in dissipation energy for both MRSA and MSSA strains. Following the bacterial binding, these sensors have been further exposed to the penicillin-binding protein antibody latex beads. Sensors analyzed with MRSA responded to PBP 2a antibody beads; although sensors tested with MSSA gave no response. This experimental distinction determines an unambiguous discrimination between methicillin resistant and sensitive S. aureus strains.

An increasing number of disease-causing bacteria are resistant to one or more anti-bacterial drugs utilized for therapy. Early and speedy detection of these pathogens is therefore very important. Traditional pathogen detection techniques, that include microbiological and biochemical assays are long and labor-intensive, while antibody or DNA-based methods require substantial sample preparation and purification. Biosensors based on bacteriophages have demonstrated remarkable potential to surmount these restrictions and to offer rapid, efficient and sensitive detection technique for antibiotic-resistant bacteria.

Background: Increased rates of locoregional recurrence have been observed in basal-like breast cancer despite the use of radiation therapy (RT), therefore approaches that result in radiosensitization of basal-like breast cancer are critically needed. Studies detailing the poor response of basal-like tumors to adjuvant RT underscore the biologic differences and as of yet undefined oncogenic drivers of these particular types of breast cancer. Methods: 4 independent datasets were used to correlate gene expression with local recurrence (LR) after 3 years. Kaplan-Meier analysis was used to validate the impact of TTK expression on LR. The TCGA and institutional breast cancer dataset were used to determine TTK expression in BC subtypes. TTK RNA and protein levels were measured using qPCR and western blot at baseline and correlated to intrinsic radiosensitivity. Clonogenic survival assays were used to determine the radiosensitization of cell lines after TTK inhibition (TTKi). Mice models were used to assess TTKi in combination with RT in vivo. DNA damage was quantified using γH2AX staining. HR and NHEJ efficiency assays were performed using HR/NHEJ specific reporter systems. HR competency was also assessed using RAD51 foci formation assays. Results: Ten genes were found to significantly correlate with early LR (≤3 years) across 4 distinct datasets (N=896 pts) (OR of recurrence > 2, p-value Conclusion: These data support TTK inhibition as a rationale radiosensitizing strategy for clinical development in basal-like breast cancer patients and that the mechanism of radiosensitization is, at least in part, through impaired HR repair. Citation Format: Benjamin C. Chandler, Leah Moubadder, Cassie Ritter, Kari Wilder-Romans, Meleah Cameron, Meilan Liu, Shyam Nyati, Andrea Pesch, Anna Michmerhuizen, Eric Olsen, Yashar Niknafs, Arul Chinnaiyan, Corey Speers. Nomination and characterization of TTK for radiosensitization in basal-like breast cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2926.

In this paper the application of poly(3,4-ethlenedioxythiophene)-poly(styrene sulphonate) (PEDOT:PSS) as an anti-fouling modification on a glassy carbon electrode has been investigated for use in an electrochemical sensing system for continuous monitoring of gaseous tricresyl phosphate (TCP). PEDOT is a type of conductive polymer with high stability in aqueous solution. The amphiphilic nature of poly(sodium-4-styrenesulfonate) (NaPSS) helps repel the oxidation products of cresol and reduces electrode fouling. The composite modification has high reproducibility which enables the quantitative determination of gaseous TCP. Although the linear range of detection (50–300 ppb) is narrower than that previously reported, this modified electrode enables continuous monitoring of TCP without the need for electrode polishing that may limit practical application of the sensor in the aircraft cabin.

In carcinogenicity studies, pregabalin increased hemangiosarcoma incidence in mice but not in rats. Investigative studies, ranging in length from 24 h to 12 months, were conducted in mice (1000 or 5000 mg/kg) and rats (900 mg/kg) to evaluate a potential mode-of-action scheme for tumor formation. Three areas were evaluated: (1) hematopoiesis (because endothelial and hematopoietic cells arise from the same precursor and hemangiosarcomas are primarily located in mouse hematopoietic tissues), (2) angiogenic growth factors (because increased angiogenic growth factors may stimulate vascular tumors), and (3) pulmonary/blood gas parameters (because hypoxia is a known driver for endothelial cell proliferation). In mice, pregabalin rapidly increased platelet and megakaryocyte counts, activated platelets and bone marrow erythrophages, decreased the myeloid-to-erythroid (M:E) ratio (49%), and produced bone marrow and splenic congestion and extramedullary hematopoiesis (EMH). Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor immunohistochemical staining were also increased in mouse bone marrow and spleen and vascular endothelial growth factor receptor 2 immunolabeling was increased in liver. Serum bicarbonate was increased within 24 h of pregabalin administration, persisted over time, and was accompanied by decreased respiratory rate (up to 34%) and increased partial pressure of carbon dioxide (pCO(2)), resulting in sustained metabolic alkalosis and elevated blood pH in mice. In contrast, in rats, pregabalin decreased overall bone marrow cellularity, including decreased number of megakaryocytes (24%) with no evidence of erythrophages, no change in M:E ratio, no EMH, and no increase in angiogenic growth factors or blood pH. Persistent alterations in serum bicarbonate, respiratory function, and blood gas parameters in mice, without adequate compensatory mechanisms, has the potential to create chronic tissue hypoxia, an accepted driver of endothelial cell proliferation.

DNA biosensors have gained increased attention over traditional diagnostic methods due to their fast and responsive operation and cost-effective design. The specificity of DNA biosensors relies on single-stranded oligonucleotide probes immobilized to a transduction platform. Here, we report the development of biosensors to detect the hippuricase gene (hipO) from Campylobacter jejuni using direct covalent coupling of thiol- and biotin-labeled single-stranded DNA (ssDNA) on both surface plasmon resonance (SPR) and diffraction optics technology (DOT, dotLab) transduction platforms. This is the first known report of the dotLab to detect targeted DNA. Application of 6-mercapto-1-hexanol as a spacer thiol for SPR gold surface created a self-assembled monolayer that removed unbound ssDNA and minimized non-specific detection. The detection limit of SPR sensors was shown to be 2.5 nM DNA while dotLab sensors demonstrated a slightly decreased detection limit of 5.0 nM (0.005 μM). It was possible to reuse the SPR sensor due to the negligible changes in sensor sensitivity (∼9.7 × 10−7 ΔRU) and minimal damage to immobilized probes following use, whereas dotLab sensors could not be reused. Results indicated feasibility of optical biosensors for rapid and sensitive detection of the hipO gene of Campylobacter jejuni using specific ssDNA as a probe.

Stable, insoluble Langmuir monolayer films composed of Staphylococcus aureus-specific lytic bacteriophage were formed at an air-water interface and characterized. The phage monolayer was very strong, withstanding a surface pressure of ∼40 mN/m at 20 °C. The surface pressure-area (Π-A) isotherm possessed a shoulder at ∼7 × 10(4)nm(2)/phage particle, attributed to a change in phage orientation at the air-water interface from horizontal to vertical capsid-down/tail-up orientation as surface pressure was increased. The Π-A-dependence was accurately described using the Volmer equation of state, assuming horizontal orientation to an air-water interface at low surface pressures with an excluded area per phage particle of 4.6 × 10(4)nm(2). At high pressures phage particles followed the space-filling densely packed disks model with a specific area of 8.5 × 10(3)nm(2)/phage particle. Lytic phage monolayers were transferred onto gold-coated silica substrates from the air-water interface at a constant surface pressure of 18 mN/m by Langmuir-Blodgett method, then dried and analyzed by scanning electron microscopy (SEM) and ellipsometry. Phage specific adsorption (Γ) in Langmuir-Blodgett (LB) films measured by SEM was consistent with that calculated independently from Π-A isotherms at the transfer surface pressure of 18 mN/m (Γ=23 phage particles/μm(2)). The 50 nm-thickness of phage monolayer measured by ellipsometer agreed well with the horizontal phage average size estimated by SEM. Surface properties of phage Langmuir monolayer compare well with other monolayers formed from nano- and micro-particles at the air-water interface and similar to that of classic amphiphiles 1,2-diphytanoyl-sn-glycero-3-phosphocholine (phospholipid) and stearic acid.

Selective lipase-catalyzed acylation of 41-desmethoxyrapamycin has been achieved with a quaternary carboxylic acid avoiding the use of vinyl ester activation. Among the acyl donors investigated, the novel butanedione-monooxime and the N-acetylhydroxamate ester proved to be the most efficient donors, comparable in reactivity to the undesired vinyl ester and allowing selective, preparative acylation on gram scale in excellent yields. These new donors are proposed as sustainable and process-friendly alternatives to the widely used vinyl ester substrate activation in lipase-catalyzed acylations of secondary alcohols.

Background: Increased rates of locoregional recurrence leading to poorer clinical outcomes have been observed in triple-negative breast cancer (TNBC) despite the use of radiation therapy (RT), therefore approaches that result in radiosensitizaton in TNBC are critically needed. Our previous work identified a group of cell cycle kinases differentially expressed in estrogen receptor negative (ER-) versus estrogen receptor positive (ER+) breast cancer (BC). Here we described the impact of elevated expression of one of these identified kinases, TTK, on radiation response and patient outcomes in TNBC. Methods: TCGA breast cancer datasets were used to determine TTK expression in the intrinsic subtypes of BC. TTK RNA and protein levels were measured using qPCR and western blot at baseline and after radiation treatment. Clonogenic survival assays were used to determine the radiosensitization in several cells after TTK inhibition. DNA damage was quantified using γH2AX staining in TNBC cell lines. Kaplan-Meier analysis was used to determine the impact of TTK expression on locoregional recurrence (LRR) and overall survival (OS). A Cox proportional hazards model was constructed to identify potential factors of LRR-free survival in univariate (UVA) and multivariable analyses (MVA). Results: TTK expression is elevated in breast cancer tissue compared to normal breast tissue (Q-VAL: 8.70 E-291) and is most highly expressed in basal-like tumors. TTK is overexpressed in ER- versus ER+ tumors in both breast cancers patient samples (p-value: Conclusion: Our results support the rationale for developing translational clinical trials to investigate TTK inhibition as a novel radiosensitizing target in TNBC. Citation Format: Benjamin C. Chandler, Leah Moubadder, Cassie Ritter, Yashar Niknafs, Eric Olsen, Meleah Cameron, Meilan Liu, Kari Wilder-Romans, Shyam Nyati, Powel Brown, Arul Chinnaiyan, Corey Speers. TTK: A novel target for radiosensitization in triple-negative breast cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 677.

Background: Increased rates of locoregional recurrence have been observed in TNBC despite the use of radiation (RT); therefore, approaches that result in radiosensitizaton in TNBC may improve clinical outcomes. Despite some controversy as to the role of MELK kinase as an oncogene, we previously established its clear role in conferring radioresistance in TNBC. Here we extend those studies to understand the mechanism of conferred radioresistance. Methods: Clonogenic survival assays were used to quantify the degree of radiosensitivity after MELK inhibition. Mass spectrometry was utilized to identify proteins associated with MELK. NHEJ assays were performed using linearized pEYFP vector and quantitated by FACS. qRT-PCR was used to determine MELK RNA expression. Rad51 and γH2AX foci counting were performed to evaluate homologous recombination (HR). Neutral comet assay was used to quantitate dsDNA break repair. Laser microirradiation studies were performed to determine the location and kinetics of protein shuttle to and from sites of dsDNA breaks, including MELK, Ku70 and Ku80 proteins. Fluorescent microscopy was performed for localization of endogenous MELK protein post RT. To study the effect of MELK kinase activity on Ku70-Ku80 complex formation and recruitment at the DNA damage site, cells were lysed and immunoprecipitation was performed using MELK, Ku70 or Ku80-specific antibodies. Results: We previously demonstrated that genomic or pharmacologic inhibition of MELK confers significant radiosensitization. Mechanistic studies were undertaken to understand the pathways critical to MELK-mediated radioresistance. When MELK is genomically or pharmacologically inhibited, NHEJ and HR reporter assays demonstrate that this radiosensitization is driven through impaired NHEJ at 4, 16, and 24 hrs after RT and not mediated by HR in multiple TNBC cell lines. Tandem mass spectrometry studies of tagged MELK protein identify Ku70 and Ku80 as direct interactors with MELK protein, which is confirmed by Co-IP. Laser microirradiation studies confirm that MELK, Ku70 and Ku80 co-localize to sites of dsDNA breaks, and that MELK kinase function is required to stabilize the Ku70/Ku80 complex at these sites. Comet assay confirms that rapid dissolution of the Ku70/Ku80 complex when MELK is inhibited or functionally dead is not a result of accelerated dsDNA break repair. Conclusion: MELK kinase function is critical in conferring radioresistance in TNBC and inhibition of function confers radiosensitivity through an NHEJ-mediated pathway. MELK also stabilizes Ku70/Ku80 proteins at sites of dsDNA breaks and allows for more efficient repair of breaks induced by ionizing RT. These results further support the rationale for developing clinical strategies to inhibit MELK in combination with RT treatment as a novel radiosensitizing strategy in TNBC. Citation Format: Shyam Nyati, Ben Chandler, Eric Olsen, Leah Moubadder, Meilan Liu, Meleah Cameron, Kari Wilder-Romans, Theodore S. Lawrence, Powel H. Brown, Fellix Y. Fang, Lori J. Pierce, Corey Speers. Maternal embryonic leucine zipper kinase (MELK) confers radioresistance in triple-negative breast cancers (TNBC) through a nonhomologous end joining (NHEJ)-mediated pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3218.

Radio Frequency Identification (RFID) is an automatic identification technology, relying on storing and remotely retrieving data using devices called RFID tags. This technology is being used in enterprise supply chain management-related applications to improve the efficiency of inventory tracking and management. However, this technology has not been able to realize its promised potential because of several factors, such as lack of congruous worldwide standards, privacy issues and less than perfect read rates in supply chain applications. This research aimed to evaluate the readability issues commonly faced by tagged cases of palletized consumer products. The variables studied in this research were product-package type, tag type, tag location on cases, pallet pattern and forklift speed through a RFID portal representative of a dock door in a warehouse. To determine which variables were the most significant, a binary logistic regression was run. The number of tags read was inputted for the number of events and total number of products per pallet as the number of trials. The variables product content, pallet pattern, and speed, and all interactions were then included in the model. It was observed that readability greatly varies for different product-package systems, with paper towels producing near-perfect reads, followed by bottled water and carbonated soda cans. The slower the forklift truck speed, the better the readability across the board, and the best pallet patterns were dependent on the product-package type. For bottled water, the best pallet pattern was column, and for carbonated soda, the interlocking pattern. Copyright © 2009 John Wiley & Sons, Ltd.

Multifunctional coatings were produced by the layer by layer assembly of single-walled carbon nanotubes (SWNT) dispersed in DNA and lysozyme (LSZ) on an insulating glass substrate. The electrochemical properties of these mechanically robust biocoatings were characterized for the first time using scanning electrochemical microscopy (SECM) and impedance spectroscopy (IS). SECM surface analysis demonstrated an increase in tip current with a corresponding increase in the number of oppositely polarized interlaced layers, indicating that subsequent layers were not electrically insulated from each other and a direct correlation exists between SECM feedback response and the number of layers. The rate of charge transport was also dependent on the chemical composition/polarity of the outermost surface layer. Coatings terminating in SWNT-DNA resulted in more positive feedback than those terminating in SWNT-LSZ. IS analysis demonstrated that the SWNT-DNA had a low charge transfer resistance in comparison with SWNT-LSZ, which is consistent with the results obtained by SECM. These results enable enhanced fundamental understanding and prediction of the electrical properties of SWNT-biopolymer layers with controlled interlaced polarities and orientation. Furthermore, these finding highlight the potential for SWNT-biopolymers in electronic and sensing applications. Keywords: Biopolymer, Scanning electrochemical microscopy, DNA, Lysozyme

Biztainment is a practice by which entertainment is added to a bundle of goods and services in order to gain competitive advantage. The achievement of this goal is illustrated herein using economic examples of increased revenue, repeat business, and profits, and by extending the product lifecycle, thus ensuring survival of the firm. The general premise is that biztainment is an increasingly popular business strategy, applicable to all industries. For example, consider the goods and services provided by Build-A-Bear stores: children can select the fabric, eyes, and buttons to create a unique tangible good, while the memorable process of building it (employing self-service, too) with family or friends adds extraordinary value to the purchase. Build-A-Bear's use of biztainment has resulted in 370 stores worldwide on five continents, expanding at a rate of 25 locations per year, and earned revenue of $474 million in 2007 (Build-A-Bear, 2008). This article concludes by offering examples of ways in which managers can evaluate their current product-service strategies against the environmental drivers of biztainment.

PurposeThis study aims to address an important gap between the normative view of an integrated performance measurement system (PMS) design that assumes a clean slate and the organizational realities of a PMS design as an ongoing analysis, coordination and improvement process.Design/methodology/approachThe authors present a framework for evaluating the effectiveness of a PMS based on three criteria – i.e. causality, continuous improvement and process control – and use a case study to illustrate the application of the methodology and the interpretation of results for PMS design.FindingsThe determination of “driver measures” in an integrated PMS involves a complex process that requires a number of considerations not adequately addressed in prior research.Research limitations/implicationsThis study involves only a single case study and the model presented involves only a two‐tier analysis.Practical implicationsThe framework provides a simple methodology that organizations can easily adopt to analyze individual and group performance measures and relate them to the strategic performance measures of the company.Originality/valueThe study follows an emerging line of research that addresses the design of an integrated PMS as an ongoing improvement process.

Filamentous phage affinity-selected for streptavidin and Salmonella typhimurium from phage display libraries were transformed with chloroform into spherical forms then deposited as phage coat protein monolayers to quartz crystal microbalances (QCM) by Langmuir-Blodgett (LB) to prepare bioselective sensors. Maximum yield of spheroids was achieved by mixing 8.3 x 10-11 to 2.5 x 10-12 filamentous phage virions/ml with 1 ml chloroform for 60 seconds. Spheroid conversion and binding to S. typhimurium was confirmed by transmission electron microscopy. Phage coat monolayers incorporating phospholipid possessed high elasticity and transference to QCM substrates comparable to antibodies. Sensor responses to increasing concentrations of target analytes were characterized by rapid reaction, steady-state equilibrium, high sensitivity, and linear dose-response that followed mass theory for piezoelectric transducers. Scanning electron microscopy confirmed binding of target analytes to biosensors compared to controls. Phage-based sensors may allow detection of bacterial agents such as S. typhimurium in food safety and security applications.

One of the principal challenges in the collection of biological samples from air, water, and soil matrices is that the target agents are not stable enough to be transferred from the collection point to the laboratory of choice without experiencing significant degradation and loss of viability. At present, there is no method to transport biological samples over considerable distances safely, efficiently, and cost-effectively without the use of ice or refrigeration. Current techniques of protection and preservation of biological materials have serious drawbacks. Many known techniques of preservation cause structural damages, so that biological materials lose their structural integrity and viability. We review applications of a novel bacterial preservation process, which is nontoxic and water soluble and allows for the storage of samples without refrigeration. The method is capable of protecting the biological sample from the effects of environment for extended periods of time and then allows for the easy release of these collected biological materials from the protective medium without structural or DNA damage. Strategies for sample collection, preservation, and shipment of bacterial, viral samples are described. The water-soluble polymer is used to immobilize the biological material by replacing the water molecules within the sample with molecules of the biopolymer. The cured polymer results in a solid protective film that is stable to many organic solvents, but quickly removed by the application of the water-based solution. The process of immobilization does not require the use of any additives, accelerators, or plastifiers and does not involve high temperature or radiation to promote polymerization.

Developing simple, reliable, and cost-effective methods of renewing an inhibited biocatalyst (e.g., enzymatic interfaces) on biosensors is needed to advance multiuse, reusable sensor applications. We report a method for the renewal of layer-by-layer (LbL) self-assembled inhibition-based enzymatic interfaces in multiwalled carbon nanotube (MWCNT) armored acetylcholinesterase (AChE) biosensors. The self-assembly process of MWCNT dispersed enzymes/biopolymers was investigated using surface plasmon resonance (SPR). The LbL fabrication consisted of alternating cushion layers of positively charged CNT-polyethylenimine (CNT-PEI) and negatively charged CNT-deoxyribonucleic acid (CNT-DNA) and a functional interface consisting of alternating layers of CNT-PEI and negatively charged CNT-acetylcholine esterase (CNT-AChE, pH 7.4). The observed SPR response signal increased while assembling the different layers, indicating the buildup of multiple layers on the Au surface. A partial desorption of the top enzymatic layer in the LbL structure was observed with a desorption strategy employing alkaline treatment. This indicates that the strong interaction of CNT-biopolymer conjugates with the Au surface was a result of both electrostatic interactions between biopolymers and the surface binding energy from CNTs: the closer the layers are to the Au surface, the stronger the interactions. In contrast, a similar LbL assembly of soluble enzyme/polyelectrolytes resulted in stronger desorption on the surface after the alkaline treatment; this led to the investigation of AChE layer removal, permanently inhibited after pesticide exposure on glassy carbon (GC) electrodes, while keeping the cushion layers intact. The desorption strategy permitted the SPR and electrochemical electrode surfaces to be regenerated multiple times by the subsequent self-assembly of fresh PEI/AChE layers. Flow-mode electrochemical amperometric analysis demonstrated good stability toward the determination of acetylcholine with 97.1 ± 2.7% renewability. Our simple, inexpensive approach shows the potential of renewable LbL self-assembled functional interfaces for multiple uses in a wide field of applications such as biosensing, various biotechnological processes, and the food and health industries.

We selected from landscape phage library probes that bind preferentially Salmonella typhimurium cells compared with other Enterobacteriaceae. The specificity of the phage probes for S. typhimurium was analyzed by the phage-capture test, the enzyme-linked immunosorbent assay (ELISA), and the precipitation test. Interaction of representative probes with S. typhimurium was characterized by fluorescence-activated cell sorting (FACS), and fluorescent, optical and electron microscopy. The results show that the landscape phage library is a rich source of specific and robust probes for S. typhimurium suitable for long-term use in continuous monitoring devices and biosorbents.

The specific and selective detection of Salmonella typhymurium based on the use of a polyclonal antibody immobilized by the Langmuir-Blodgett method on the surface of a quartz crystal acoustic wave device was demonstrated in liquid samples. These biosensors were selective to S. typhymurium in the presence of large concentrations of Escherichia coli O157:H7. They were also specific to S. typhymurium since bacteria preincubated with free antibody produced no signal. Dark-field and electron microscopy showed that two different antibodies, polyvalent somatic O and flagellar H7, were immobilized on the sensor surface producing two distinct attachments of bacteria at the liquid-solid interface. The somatic O antibody exhibits a rigid, binding, while the flagellar H7 antibody forms a flexible connection allowing a large degree of freedom. When the attachment of bacteria was rigid and strong, the responses of the acoustic wave sensors correlated with changes in the mass of bacteria present at the liquid-solid interface. In contrast, when attachment was flexible, the sensor signals were inversely proportional to the additional mass of bound bacteria. This difference is probably determined by the interfacial viscoelasticity and by acoustic and electromagnetic coupling. The signals of environmentally aged sensors with either predominantly rigid or flexible positioning of bacteria were correlated with changes in mass at the liquid-solid interface. Sensors with O or H type of binding could be used for analytical purposes.

Value‐based management theories have fallen short in practice, especially when it comes to investor strategy.

Purpose: Increased rates of local recurrence (LR) have been observed in TNBC despite chemotherapy and radiation (RT). Thus, approaches that result in radiosensitization in TNBC are critically needed. We characterized the RT response of 21 breast cancer cell (BCC) lines using clonogenic survival assays and paired this with high-throughput drug screen data, identifying AR as a top target for radiosensitization. We demonstrate that AR inhibition confers radiosensitization in vitro and in vivo, identified a biomarker of response, and characterize the mechanism of AR-mediated radiosensitization in TNBC. Materials/Methods: Clonogenic survival assays determined the intrinsic RT sensitivity of 21 BCC lines. IC50 values were determined for 130 clinical compounds and correlation coefficients were calculated using IC50 values and SF-2Gy. Gene and protein expression was measured using RNA Seq and RPPA arrays, respectively, in tumor samples (n=2,061) and BCC lines (n=51). AR function was assessed using gene knockdown (KD) or functional inhibition with anti-androgen drugs. We measured in vivo tumor growth with varying control and treatment groups (16-20 tumors/group). Kaplan-Meier analysis was performed to estimate local control. A Cox proportional hazards model and MVA were used to determine variables associated with LRF survival. Results: Our unbiased drug radiosensitizer screen nominated bicalutamide as an effective drug in treating RT-resistant BCC lines (R2= 0.46, p-value 2000 human breast tumor samples and 51 BCC lines and found heterogeneity in AR expression with strongly correlated expression of protein and RNA levels in TNBC (R2=0.89, p-value Conclusion: Our results implicate AR as a mediator of radioresistance in breast cancer and support the rationale for developing clinical strategies, including clinical trials, to inhibit AR as a novel radiosensitizing target in TNBC. Citation Format: Benjamin C. Chandler, Corey W. Speers, Shuang G. Zhao, Meilan Liu, Kari Wilder-Romans, Eric Olsen, Shyam Nyati, Daniel Spratt, Daniel Wahl, Daniel Hayes, Felix Y. Feng, Lori J. Pierce. Androgen receptor (AR): A novel target for radiosensitization in triple-negative breast cancers (TNBC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5839. doi:10.1158/1538-7445.AM2017-5839

Tumor-associated macrophages play an important role in presenting tumor antigens to T cells and are being investigated for their role in sensitivity to immune checkpoint inhibitors such as PD-1 inhibitors. The M2 macrophage subtype, characterized by CD163 expression, is immunosuppressive and associated with poor prognosis. A multiplex CD68/CD163 immunohistochemical assay was developed to characterize the density of cells expressing each marker and determine the percentage of macrophages that are of the M2 subtype. The assay was used to analyze 22 normal tissue types and 50 tumor tissues within 5 different indications. The results were analyzed by image analysis and results were analyzed as percent positive and density per mm2. Cancer tissues expressed a greater percentage of CD163+ macrophages than normal tissues. Citation Format: Lisa M. Dauffenbach, Gela C. Sia, Jianping Zheng, Natalia Jun, Eric P. Olsen, Christopher A. Kerfoot. Characterization of M2 macrophage subtype content in cancer and normal tissues [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4009. doi:10.1158/1538-7445.AM2017-4009