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Background: HER2-targeted therapies have revolutionised the treatment of HER2-positive breast cancer. However, de novo resistance or the emergence of acquired resistance is a persistent clinical problem. Here we report that neratinib, an irreversible pan-HER inhibitor, in combination with the multi-kinase inhibitor dasatinib, currently used to treat certain leukemias, has strong anti-proliferative effects against models of HER2-positive breast cancer that are innately resistant to trastuzumab or have acquired resistance to neratinib., Methods: Neratinib plus dasatinib was examined in a panel of 20 breast cancer cell lines, including HER2-positive, estrogen-receptor-positive, triple negative, and acquired HER2-targeted therapy resistant models. Drug effects on migration and apoptosis induction was evaluated and signaling alterations were determined by reverse phase protein array (RPPA). In vivo efficacy was examined using orthotopically-implanted HCC1954 cells., Results: Synergy was observed in cell lines innately resistant to trastuzumab, models with acquired resistance to neratinib, and in triple negative breast cancer cell lines. Further investigation showed that neratinib plus dasatinib induced apoptosis and inhibited cell migration to a greater degree than either drug alone. RPPA revealed that the combination caused suppression of key survival signaling through EGFR, Akt, and MAPK inhibition. In vivo, neratinib plus dasatinib was well tolerated and had a prolonged anti-tumor effect against HCC1954 xenografts., Conclusions: This study provides a strong pre-clinical rationale for the clinical investigation neratinib and dasatinib in HER2+ breast cancer., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: The conflict of interest for this study are that DMC, NTC and JC have received academic research funding from Puma Biotechnology Inc., DMC has received consultancy fees from Puma Biotechnology Inc. JC has been on advisory boards for Puma Biotechnology Inc, Seattle Genetics, and Novartis Ireland. LDE is an employee and shareholder in Puma Biotechnology Inc. All remaining authors have no conflicts of interest to declare., (Copyright © 2024. Published by Elsevier Inc.)

CDS enzymes (CDS1 and 2 in mammals) convert phosphatidic acid (PA) to CDP-DG, an essential intermediate in the de novo synthesis of PI. Genetic deletion of CDS2 in primary mouse macrophages resulted in only modest changes in the steady-state levels of major phospholipid species, including PI, but substantial increases in several species of PA, CDP-DG, DG and TG. Stable isotope labelling experiments employing both 13C6- and 13C6D7-glucose revealed loss of CDS2 resulted in a minimal reduction in the rate of de novo PI synthesis but a substantial increase in the rate of de novo PA synthesis from G3P, derived from DHAP via glycolysis. This increased synthesis of PA provides a potential explanation for normal basal PI synthesis in the face of reduced CDS capacity (via increased provision of substrate to CDS1) and increased synthesis of DG and TG (via increased provision of substrate to LIPINs). However, under conditions of sustained GPCR-stimulation of PLC, CDS2-deficient macrophages were unable to maintain enhanced rates of PI synthesis via the 'PI cycle', leading to a substantial loss of PI. CDS2-deficient macrophages also exhibited significant defects in calcium homeostasis which were unrelated to the activation of PLC and thus probably an indirect effect of increased basal PA. These experiments reveal that an important homeostatic response in mammalian cells to a reduction in CDS capacity is increased de novo synthesis of PA, likely related to maintaining normal levels of PI, and provides a new interpretation of previous work describing pleiotropic effects of CDS2 deletion on lipid metabolism/signalling., (© 2024 The Author(s).)

This study describes the development and content validation of the Electronic Instrumental activities of daily living Satisfaction Assessment (EISA), a self-report outcome measure for assessing satisfaction with completing instrumental Activities of Daily Living (IADL) for People With Disabilities (PWD) using internet-Connected Assistive Devices (iCAD). For this study, an iCAD is defined as any information communication technology/electronic device or software that assists with promoting, maintaining, and/or enhancing the ability of a PWD to live independently in society. Phase 1 of development involved generating an initial item pool based on a literature review. Content validity of the EISA was computed in Phase 2 using the Qualtrics on-line research platform. Utilizing the content validity index procedure, EISA demonstrated acceptable content validity: item level (I-CVI of 0.78 or higher) and scale level (S-CVI/Ave of 0.90 or higher). EISA version 1.0 was generated in Phase 3 where it is the first of its kind outcome measure specifically designed for assessing satisfaction with completing IADL for PWD using iCAD.

Purpose Empathetic engagement is considered a vital component in forming respect-based relationships between patients and clinicians, leading to more optimal patient care. The purpose of this study was to explore whether there was a relationship between dental hygiene students' levels of empathy and student demographics including age, gender, year in school, and the degree type of dental hygiene program attending. Methods This was a cross-sectional observation study conducted among dental hygiene undergraduate students attending three dental hygiene programs in the Midwestern United States. Two programs offered associate degrees and one offered a baccalaureate degree. Participants completed the 20-item Jefferson Scale of Empathy © , student edition (JSE-S) along with demographic questions including age, gender, year in dental hygiene program, and degree type of dental hygiene program. Descriptive statistics and comparisons of the empathy scores were conducted using t-test and one-way analysis of variance (ANOVA). Regressions were conducted to determine whether the students' year in dental hygiene program and the type of degree program were predictors of empathy. Results Forty-one participants completed the questionnaire for a 65% response rate. The mean empathy score was 83.05 ± 10.04 among the participants. There were no statistically significant differences between levels of empathy of first- and second-year students or those attending a two-year institution versus a four-year university. Age, year in program, and type of degree were not shown to be predictors of empathy. Conclusion Results from this study did not show relationships or predictors of empathy with dental hygiene students' demographics or type of degree program. Future research should expand beyond a small homogenous convenience sample and include a longitudinal gauge to assess potential fluctuations in empathy as students progress throughout the curriculum and as practicing clinicians., (Copyright © 2024 The American Dental Hygienists’ Association.)

Purpose: Drug efflux transporter associated multi-drug resistance (MDR) is a potential limitation in the use of taxane chemotherapies for the treatment of metastatic melanoma. ABT-751 is an orally bioavailable microtubule-binding agent capable of overcoming MDR and proposed as an alternative to taxane-based therapies., Methods: This study compares ABT-751 to taxanes in vitro, utilizing seven melanoma cell line models, publicly available gene expression and drug sensitivity databases, a lung cancer cell line model of MDR drug efflux transporter overexpression (DLKP-A), and drug efflux transporter ATPase assays., Results: Melanoma cell lines exhibit a low but variable protein and RNA expression of drug efflux transporters P-gp, BCRP, and MDR3. Expression of P-gp and MDR3 correlates with sensitivity to taxanes, but not to ABT-751. The anti-proliferative IC 50 profile of ABT-751 was higher than the taxanes docetaxel and paclitaxel in the melanoma cell line panel, but fell within clinically achievable parameters. ABT-751 IC 50 was not impacted by P-gp-overexpression in DKLP-A cells, which display strong resistance to the P-gp substrate taxanes compared to DLKP parental controls. The addition of ABT-751 to paclitaxel treatment significantly decreased cell proliferation, suggesting some reversal of MDR. ATPase activity assays suggest that ABT-751 is a potential BCRP substrate, with the ability to inhibit P-gp ATPase activity., Conclusion: Our study confirms that ABT-751 is active against melanoma cell lines and models of MDR at physiologically relevant concentrations, it inhibits P-gp ATPase activity, and it may be a BCRP and/or MDR3 substrate. ABT-751 warrants further investigation alone or in tandem with other drug efflux transporter inhibitors for hard-to-treat MDR melanoma., (© 2024. The Author(s).)

Background: Pressure injuries are one of the most challenging secondary conditions for individuals with spinal cord injuries and related disorders (SCI/D) owing to inherent, lifelong risk factors that include a lack of sensory and motor function below the level of injury and reliance on a wheelchair for daily mobility, resulting in prolonged periods of sitting. Although many factors contribute to the development of pressure injuries, the pressure between the skin and a surface is always a factor and the development of injury is dependent on the magnitude and duration of the pressure. Clinically, broad recommendations for relieving pressure are used because we know very little about the unique day-to-day life patterns of the individual wheelchair user. Typically, it is after the occurrence of a pressure injury that the therapist will check equipment fit and the effectiveness of pressure offloading and ask about other surfaces they sit on in their home and community. This time-lapsed, largely self-reported data are fraught with recall bias and inaccuracies that the therapist incorporates into a plan of care., Objective: This study's objective is to pilot-test the implementation and clinical effectiveness of a telehealth model of care combined with our mobile health (mHealth) Assisted Weight-Shift device for remote monitoring of factors related to maintaining skin health and wheelchair setup. Our overall hypothesis is that this study will result in an effective implementation plan, and the enhanced connected model of care using remote monitoring of pressure management will result in pilot-level, improved clinical outcomes for adults with spinal cord injury at high risk for pressure injury recurrence., Methods: For all aims, we will use a mixed methods design using an exploratory, sequential approach to include the strengths of both qualitative and quantitative data. For aims 1 and 2, we will iteratively collect qualitative data from therapists, patients with SCI/D, and other stakeholders. For aim 3, we will perform a hybrid effectiveness-implementation randomized controlled trial to pilot-test the intervention. The projected results include an iteratively developed and tested implementation plan that meets moderate to high levels of acceptability, feasibility, and appropriateness. Additionally, the pilot trial results are expected to show positive trends in relevant clinical outcomes related to reduced pressure injury incidence, recurrence, and improved healing when compared with the standard of care., Results: Currently, 6 participants have been recruited for our aim-1 qualitative study., Conclusions: This study will expand upon our previous study to move the Assisted Weight-Shift system into routine clinical care, which was a strong desire of adults with SCI/D for improved individualized care plans to prevent pressure injuries. The results of this study will guide the next steps in a full, hybrid effectiveness-implementation trial with the goal of improving care to prevent pressure injuries., International Registered Report Identifier (irrid): DERR1-10.2196/51849., (©Melissa M Morrow, Lynne C Hughes, Diane M Collins, Tamara L Vos-Draper. Originally published in JMIR Research Protocols (https://www.researchprotocols.org), 10.04.2024.)

Over the past decade, the Diagnostic and Statistical Manual 's method of prescribing medications based on presenting symptoms has been challenged. The shift toward precision medicine began with the National Institute of Mental Health and culminated with the World Psychiatric Association's posit that a paradigm shift is needed. This study supports that shift by providing evidence explaining the high rate of psychiatric medication failure and suggests a possible first step toward precision medicine. A large psychiatric practice began collecting electroencephalograms (EEGs) for this study in 2012. The EEGs were analyzed by the same neurophysiologist (board certified in electroencephalography) on 1,233 patients. This study identified 4 EEG biomarkers accounting for medication failure in refractory patients: focal slowing, spindling excessive beta, encephalopathy, and isolated epileptiform discharges. Each EEG biomarker suggests underlying brain dysregulation, which may explain why prior medication attempts have failed. The EEG biomarkers cannot be identified based on current psychiatric assessment methods, and depending upon the localization, intensity, and duration, can all present as complex behavioral or psychiatric issues. The study highlights that the EEG biomarker identification approach can be a positive step toward personalized medicine in psychiatry, furthering the clinical thinking of "testing the organ we are trying to treat.", Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Importance: Currently, no self-report instruments exist for assessing satisfaction with performing instrumental activities of daily living and occupations for people with disabilities using internet-connected assistive devices like accessible smartphones, tablets, laptops, and apps., Objective: To assess the test-retest reliability and internal consistency of the Electronic Instrumental activities of daily living Satisfaction Assessment (EISA) self-report outcome tool., Design: Repeated-measures cohort study with a time frame of 7 to 21 days., Setting: Multicity online recruitment at assistive technology clinics, nongovernmental organizations, advocacy and peer support groups for people with disabilities, and higher education institutions., Participants: Eighty-four participants with disabilities, age 18 yr or older, with a mean age of 43.3 yr (range = 19-75 yr), and 57% female., Intervention: Not applicable., Outcomes and Measures: The a priori study hypotheses were that the EISA test-retest reliability scores would be above the minimum acceptable level (Rs > .80) and that internal consistency would be good (Cronbach's α = .70-.90)., Results: On the basis of the study data, the EISA, Version 1.0, demonstrated good test-retest reliability (Rs = .81) and excellent internal consistency (Cronbach's α = .88)., Conclusions and Relevance: The results of the test-retest reliability and internal consistency analyses provide good support for the EISA to be used in clinical settings. What This Article Adds: This article documents the reliability and internal consistency of, to our knowledge, the first-ever self-report instrument for assessing satisfaction with performance of everyday occupations for people with disabilities using internet-connected assistive devices such as smartphones, tablets, laptops, and apps., (Copyright © 2023 by the American Occupational Therapy Association, Inc.)

Background: The phase II neo-adjuvant clinical trial ICORG10-05 (NCT01485926) compared chemotherapy in combination with trastuzumab, lapatinib or both in patients with HER2+ breast cancer. We studied circulating immune cells looking for alterations in phenotype, genotype and cytotoxic capacity (direct and antibody-dependent cell-mediated cytotoxicity (ADCC)) in the context of treatment response., Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from pre- (n = 41) and post- (n = 25) neo-adjuvant treatment blood samples. Direct/trastuzumab-ADCC cytotoxicity of patient-derived PBMCs against K562/SKBR3 cell lines was determined ex vivo. Pembrolizumab was interrogated in 21 pre-treatment PBMC ADCC assays. Thirty-nine pre-treatment and 21 post-treatment PBMC samples were immunophenotyped. Fc receptor genotype, tumour infiltrating lymphocyte (TIL) levels and oestrogen receptor (ER) status were quantified., Results: Treatment attenuated the cytotoxicity/ADCC of PBMCs. CD3+/CD4+/CD8+ T cells increased following therapy, while CD56+ NK cells/CD14+ monocytes/CD19+ B cells decreased with significant post-treatment immune cell changes confined to patients with residual disease. Pembrolizumab-augmented ex vivo PBMC ADCC activity was associated with residual disease, but not pathological complete response. Pembrolizumab-responsive PBMCs were associated with lower baseline TIL levels and ER+ tumours., Conclusions: PBMCs display altered phenotype and function following completion of neo-adjuvant treatment. Anti-PD-1-responsive PBMCs in ex vivo ADCC assays may be a biomarker of treatment response., (© 2023. The Author(s).)

Background: This novel study forms part of a larger research programme seeking an improved understanding of aspects of the owned dog population in Ireland. Dog welfare organisations (DWOs) in Ireland are recognised as an instrumental pillar of the animal welfare sector with some receiving substantial public funding. We conducted a survey of DWOs in Ireland (n = 39) to gain a better understanding of their role and function, including their policies and procedures and the rehoming of dogs to other regions. In addition, we wanted to get a better understanding of the challenges experienced by DWOs in fulfilling their role and their perspectives on potential solutions to these challenges. The survey questions consisted of closed and open-ended items. Closed items were analysed quantitively; open-ended items were analysed thematically., Results: Most DWOs (> 80%) had written protocols for important welfare actions including rehoming procedures, assessment of owner suitability and euthanasia. DWOs sent dogs to Northern Ireland (13%), Great Britain (38.5%) and to other countries outside the United Kingdom (36%, including Germany, Sweden, Italy, the Netherlands and Czechia). Reported challenges included a general lack of funding, limited public awareness of the importance of dog welfare and insufficient capacity to handle dog numbers. To address these challenges, the DWOs highlighted the potential contribution of subsidised programmes and access to resources to educate potential owners. In a further qualitative evaluation to capture perceptions of appropriate solutions by DWOs, several themes emerged, relating to legislation, education, an overwhelmed workforce, and funding., Conclusions: This study provides important insights into the roles and functions of DWOs and challenges they experience in Ireland. It is hoped that the findings from this research will inform future research investigating potential solutions to these challenges as well as the development of policy in Ireland., (© 2023. Veterinary Ireland.)

Triple Negative Breast Cancer (TNBC), a subtype of breast cancer, has fewer successful therapeutic therapies than other types of breast cancer. Insulin-like growth factor receptor 1 (IGF1R) and the Insulin receptor (IR) are associated with poor outcomes in TNBC. Targeting IGF1R has failed clinically. We aimed to test if inhibiting both IR/IGF1R was a rationale therapeutic approach to treat TNBC. We showed that despite IGF1R and IR being expressed in TNBC, their expression is not associated with a negative survival outcome. Furthermore, targeting both IR/IGF1R with inhibitors in multiple TNBC cell lines did not inhibit cell growth. Linsitinib, a small molecule inhibitor of both IGF1R and IR, did not block tumour formation and had no effect on tumour growth in vivo. Cumulatively these data suggest that while IGF1R and IR are expressed in TNBC, they are not good therapeutic targets. A potential reason for the limited anti-cancer impact when IR/IGF1R was targeted may be because multiple signalling pathways are altered in TNBC. Therefore, targeting individual signalling pathways may not be sufficient to inhibit cancer growth., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Roche et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Phosphoinositides (PIPn) in mammalian tissues are enriched in the stearoyl/arachidonoyl acyl chain species ("C38:4"), but its functional significance is unclear. We have used metabolic tracers (isotopologues of inositol, glucose and water) to study PIPn synthesis in cell lines in which this enrichment is preserved to differing relative extents. We show that PIs synthesised from glucose are initially enriched in shorter/more saturated acyl chains, but then rapidly remodelled towards the C38:4 species. PIs are also synthesised by a distinct 're-cycling pathway', which utilises existing precursors and exhibits substantial selectivity for the synthesis of C38:4-PA and -PI. This re-cycling pathway is rapidly stimulated during receptor activation of phospholipase-C, both allowing the retention of the C38:4 backbone and the close coupling of PIPn consumption to its resynthesis, thus maintaining pool sizes. These results suggest that one property of the specific acyl chain composition of PIPn is that of a molecular code, to facilitate 'metabolic channelling' from PIP2 to PI via pools of intermediates (DG, PA and CDP-DG) common to other lipid metabolic pathways., (© 2022 The Authors. Published under the terms of the CC BY 4.0 license.)

Background: Reliable information about national pet dog populations is an important contributor to informed decision-making, both by governments and national dog welfare organisations. In some countries, there is an improved understanding of aspects of the national pet dog population, but as yet limited published information is available in Ireland. The current study reviews the utility of existing data to inform our understanding of recent changes to the pet dog population in Ireland, including both biological and organisational processes., Results: Based on national data on dog licencing and microchipping registration, pet dog numbers have remained relatively stable in recent years (ie prior to the COVID-19 pandemic). Since 2015, there has been a substantial decrease in the number of dogs managed through dog control centres. Although the completeness of the data are likely variable, there appears to be substantial, and increasing, number of dogs moving from Ireland to other countries, including UK, Sweden, Italy, Germany and Singapore. We also note an increase (albeit much smaller) in the number of dogs being moved into Ireland., Conclusions: This study highlights the challenges faced when using existing national data to gain insights into the dog population of Ireland. The linking of existing national databases (individual dog identification, dog licencing, dog control statistics) has the potential to improve both the representativeness and accuracy of information about the Irish pet dog population. In the next phases of our work, we will focus on the work of dog welfare organisations, given both the increased role played by these organisations and the substantial public funding that has been committed in this sector., (© 2022. The Author(s).)

Three-dimensional X-ray diffraction (3DXRD) is shown to be feasible at the I12 Joint Engineering, Environmental and Processing (JEEP) beamline of Diamond Light Source. As a demonstration, a microstructually simple low-carbon ferritic steel was studied in a highly textured and annealed state. A processing pipeline suited to this beamline was created, using software already established in the 3DXRD user community, enabling grain centre-of-mass positions, orientations and strain tensor elements to be determined. Orientations, with texture measurements independently validated from electron backscatter diffraction (EBSD) data, possessed a ∼0.1° uncertainty, comparable with other 3DXRD instruments. The spatial resolution was limited by the far-field detector pixel size; the average of the grain centre of mass position errors was determined as ±∼80 µm. An average per-grain error of ∼1 × 10 -3 for the elastic strains was also measured; this could be reduced in future experiments by improving sample preparation, geometry calibration, data collection and analysis techniques. Application of 3DXRD onto I12 shows great potential, where its implementation is highly desirable due to the flexible, open architecture of the beamline. User-owned or designed sample environments can be used, thus 3DXRD could be applied to previously unexplored scientific areas., (open access.)

This review focuses on immune checkpoint inhibitors - immunomodulatory agents that aim to relieve tumour-mediated immune-cell suppression. Immune checkpoint proteins can be expressed on the tumour-cell or immune-cell populations. Immune checkpoint proteins dampen the immune response by inactivating immune cells capable of tumour destruction. Blockade of immune checkpoints has shown impressive results in a range of solid cancers, particularly melanoma and non-small cell lung cancer. The potential benefit of this class of drugs is widespread across most cancer types and an unprecedented number of clinical studies are underway to examine the benefit of these agents. The aims of this review are to: provide an overview of the key early immune checkpoint inhibitor trials involving drugs targeting programmed cell death-1 (PD-1), programmed cell death ligand-1 (PD-L1) and cytotoxic T-lymphocyte antigen-4 (CTLA-4) in multiple disease types; provide an overview of emerging therapies aimed at these targets; and provide a detailed exploration of the status of immune checkpoint inhibitors in breast cancer., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Sperm characteristics, such as sperm morphology and sperm morphometry are important in assessing sperm quality. This is especially important for the management and conservation of endangered and exotic species, like the Florida manatee, where information of this nature is extremely limited. In this study, we fill this knowledge gap to better understand the reproductive physiology of Florida manatees by conducting the first extensive analysis of sperm morphometry and ultrastructure. Sperm were retrieved from the vas deferens of nine recently deceased Florida manatees. Computer-aided sperm morphology analysis (CASMA) was used for morphometric analysis and laser-scanning confocal microscopy and electron microscopy were used for structural and ultrastructural characterization. Our findings reveal new morphometric and structural data for the Florida manatee spermatozoon. Twelve morphometric features of Florida manatee sperm were quantified with some approximately 1.5-2 times larger than those previously reported. Ultrastructurally, the Florida manatee spermatozoon followed a mammalian structural pattern with an ovate-shaped head, midpiece containing 84-90 mitochondria, and a flagellum. However, unique ultrastructural features were identified. Distinct, rectangular-like enlargement of four outer dense fibers surrounding the axoneme was evident, which may provide additional tensile strength to counteract the forces on sperm transiting the female reproductive tract. Likewise, strong localization of F-actin fibers within the midpiece may function to maintain sperm integrity within the female reproductive tract. These findings highlight the potential effects of sexual selective pressures on sperm size and structure in the Florida manatee and provide avenues for research on the occurrence of sperm competition in this species., (© 2021 The Authors. The Anatomical Record published by Wiley Periodicals LLC on behalf of American Association for Anatomy.)

Chronic disease management coupled with education may improve outcomes for those with chronic disease. As Human Immunodeficiency Virus (HIV) is now a chronic disease, addressing the needs of as people living with HIV (PLWH) is essential as they experience an accelerated aging process due to the mechanisms of the disease and medications taken. Although studies exist on the management of chronic HIV, few discuss the implications of medication adherence and activities of daily living related to falls among PLWH. To inform occupational therapy services for PLWH, this case-control study used extracted data from the electronic medical records of PLWH who had received occupational therapy (OT) at a large academic hospital. Two-hundred-and-four subjects were included in the final dataset; sixty-eight were cases that reported a fall within the last 12 months, while 136 were controls which were PLWH who had not sustained a fall. The association between falls and antiretroviral therapy adherence indicated males who were ART non-adherent and had balance deficits were more likely to fall. The association between ADL dysfunction and falls among PLWH showed those more likely to fall had moderate ADL dysfunction and balance deficits. The findings suggest further examination of the person factors of PLWH who are categorized clinically as non-adherent with antiretroviral therapy and have ADL dysfunction may improve health outcomes and reduce falls when paired with occupation-based interventions.

Background: Pre-treatment tumour-associated lymphocytes (TILs) and stromal lymphocytes (SLs) are independent predictive markers of future pathological complete response (pCR) in HER2-positive breast cancer. Whilst studies have correlated baseline lymphocyte levels with subsequent pCR, few have studied the impact of neoadjuvant therapy on the immune environment., Methods: We performed TIL analysis and T-cell analysis by IHC on the pretreatment and 'On-treatment' samples from patients recruited on the Phase-II TCHL (NCT01485926) clinical trial. Data were analysed using the Wilcoxon signed-rank test and the Spearman rank correlation., Results: In our sample cohort (n = 66), patients who achieved a pCR at surgery, post-chemotherapy, had significantly higher counts of TILs (p = 0.05) but not SLs (p = 0.08) in their pre-treatment tumour samples. Patients who achieved a subsequent pCR after completing neo-adjuvant chemotherapy had significantly higher SLs (p = 9.09 × 10 -3 ) but not TILs (p = 0.1) in their 'On-treatment' tumour biopsies. In a small cohort of samples (n = 16), infiltrating lymphocyte counts increased after 1 cycle of neo-adjuvant chemotherapy only in those tumours of patients who did not achieve a subsequent pCR. Finally, reduced CD3 + (p = 0.04, rho = 0.60) and CD4 + (p = 0.01, rho = 0.72) T-cell counts in 'On-treatment' biopsies were associated with decreased residual tumour content post-1 cycle of treatment; the latter being significantly associated with increased likelihood of subsequent pCR (p < 0.01)., Conclusions: The immune system may be 'primed' prior to neoadjuvant treatment in those patients who subsequently achieve a pCR. In those patients who achieve a pCR, their immune response may return to baseline after only 1 cycle of treatment. However, in those who did not achieve a pCR, neo-adjuvant treatment may stimulate lymphocyte influx into the tumour.

The Na/K-ATPase is the specific receptor for cardiotonic steroids (CTS) such as ouabain and digoxin. At pharmacological concentrations used in the treatment of cardiac conditions, CTS inhibit the ion-pumping function of Na/K-ATPase. At much lower concentrations, in the range of those reported for endogenous CTS in the blood, they stimulate hypertrophic growth of cultured cardiac myocytes through initiation of a Na/K-ATPase-mediated and reactive oxygen species (ROS)-dependent signaling. To examine a possible effect of endogenous concentrations of CTS on cardiac structure and function in vivo, we compared mice expressing the naturally resistant Na/K-ATPase α1 and age-matched mice genetically engineered to express a mutated Na/K-ATPase α1 with high affinity for CTS. In this model, total cardiac Na/K-ATPase activity, α1, α2, and β1 protein content remained unchanged, and the cardiac Na/K-ATPase dose-response curve to ouabain shifted to the left as expected. In males aged 3-6 months, increased α1 sensitivity to CTS resulted in a significant increase in cardiac carbonylated protein content, suggesting that ROS production was elevated. A moderate but significant increase of about 15% of the heart-weight-to-tibia-length ratio accompanied by an increase in the myocyte cross-sectional area was detected. Echocardiographic analyses did not reveal any change in cardiac function, and there was no fibrosis or re-expression of the fetal gene program. RNA sequencing analysis indicated that pathways related to energy metabolism were upregulated, while those related to extracellular matrix organization were downregulated. Consistent with a functional role of the latter, an angiotensin-II challenge that triggered fibrosis in the α1 r/r α2 s/s mouse failed to do so in the α1 s/s α2 s/s . Taken together, these results are indicative of a link between circulating CTS, Na/K-ATPase α1, ROS, and physiological cardiac hypertrophy in mice under baseline laboratory conditions.

Background: Human epidermal growth factor 2 (HER2/ERBB2) is frequently amplified/mutated in cancer. The tyrosine kinase inhibitors (TKIs) lapatinib, neratinib, and tucatinib are FDA-approved for the treatment of HER2-positive breast cancer. Direct comparisons of the preclinical efficacy of the TKIs have been limited to small-scale studies. Novel biomarkers are required to define beneficial patient populations., Methods: In this study, the anti-proliferative effects of the three TKIs were directly compared using a 115 cancer cell line panel. Novel TKI response/resistance markers were identified through cross-analysis of drug response profiles with mutation, gene copy number and expression data., Results: All three TKIs were effective against HER2-amplified breast cancer models; neratinib showing the most potent activity, followed by tucatinib then lapatinib. Neratinib displayed the greatest activity in HER2-mutant and EGFR-mutant cells. High expression of HER2, VTCN1, CDK12, and RAC1 correlated with response to all three TKIs. DNA damage repair genes were associated with TKI resistance. BRCA2 mutations were correlated with neratinib and tucatinib response, and high expression of ATM, BRCA2, and BRCA1 were associated with neratinib resistance., Conclusions: Neratinib was the most effective HER2-targeted TKI against HER2-amplified, -mutant, and EGFR-mutant cell lines. This analysis revealed novel resistance mechanisms that may be exploited using combinatorial strategies.

Purpose: Antibody-dependent cell-mediated cytotoxicity (ADCC) is one mechanism of action of the monoclonal antibody (mAb) therapies trastuzumab and pertuzumab. Tyrosine kinase inhibitors (TKIs), like lapatinib, may have added therapeutic value in combination with mAbs through enhanced ADCC activity. Using clinical data, we examined the impact of lapatinib on HER2/EGFR expression levels and natural killer (NK) cell gene signatures. We investigated the ability of three TKIs (lapatinib, afatinib, and neratinib) to alter HER2/immune-related protein levels in preclinical models of HER2-positive (HER2 + ) and HER2-low breast cancer, and the subsequent effects on trastuzumab/pertuzumab-mediated ADCC., Experimental Design: Preclinical studies (proliferation assays, Western blotting, high content analysis, and flow cytometry) employed HER2 + (SKBR3 and HCC1954) and HER2-low (MCF-7, T47D, CAMA-1, and CAL-51) breast cancer cell lines. NCT00524303 provided reverse phase protein array-determined protein levels of HER2/pHER2/EGFR/pEGFR. RNA-based NK cell gene signatures (CIBERSORT/MCP-counter) post-neoadjuvant anti-HER2 therapy were assessed (NCT00769470/NCT01485926). ADCC assays utilized flow cytometry-based protocols., Results: Lapatinib significantly increased membrane HER2 levels, while afatinib and neratinib significantly decreased levels in all preclinical models. Single-agent lapatinib increased HER2 or EGFR levels in 10 of 11 (91%) tumor samples. NK cell signatures increased posttherapy ( P = 0.03) and associated with trastuzumab response ( P = 0.01). TKI treatment altered mAb-induced NK cell-mediated ADCC in vitro , but it did not consistently correlate with HER2 expression in HER2 + or HER2-low models. The ADCC response to trastuzumab and pertuzumab combined did not exceed either mAb alone., Conclusions: TKIs differentially alter tumor cell phenotype which can impact NK cell-mediated response to coadministered antibody therapies. mAb-induced ADCC response is relevant when rationalizing combinations for clinical investigation., (©2020 American Association for Cancer Research.)

Internet-Connected Assistive Devices (iCAD), like accessible smartphones, tablets, computers, and apps, have become an integral part of everyday functioning for people with disabilities (PWD). The objective of this article was to identify self-report assessments having the relevance and clinical applicability for assessing satisfaction with performing Instrumental Activities of Daily Living (IADL) for PWD using iCAD. An exploratory review was conducted to identify self-report assessments that were appropriate, practical, clinically fit, and psychometrically acceptable, for assessing satisfaction with performing IADLs for PWD using iCAD. Thirty-two IADL assessments were identified, of which six met inclusion criteria. Four of six assessments did not specifically address iCAD usage, while two assessments had limited relevance and clinical applicability, for assessing satisfaction with performing IADLs for PWD using iCAD. This review establishes the growing need for a self-report IADL assessment that has been specifically designed and validated for assessing satisfaction with performing IADLs for PWD using iCAD.

The PI3Kγ isoform is activated by Gi-coupled GPCRs in myeloid cells, but the extent to which the two endogenous complexes of PI3Kγ, p101/p110γ and p84/p110γ, receive direct regulation through Gβγ or indirect regulation through RAS and the sufficiency of those inputs is controversial or unclear. We generated mice with point mutations that prevent Gβγ binding to p110γ (RK552DD) or to p101 (VVKR777AAAA) and investigated the effects of these mutations in primary neutrophils and in mouse models of neutrophilic inflammation. Loss of Gβγ binding to p110γ substantially reduced the activation of both p101/p110γ and p84/p110γ in neutrophils by various GPCR agonists. Loss of Gβγ binding to p101 caused more variable effects, depending on both the agonist and cellular response, with the biggest reductions seen in PIP 3 production by primary neutrophils in response to LTB4 and MIP-2 and in the migration of neutrophils during thioglycolate-induced peritonitis or MIP2-induced ear pouch inflammation. We also observed that p101 VVKR777AAAA neutrophils showed enhanced p84-dependent ROS responses to f MLP and C5a, suggesting that competition may exist between p101/p110γ and p84/p110γ for Gβγ subunits downstream of GPCR activation. GPCRs did not activate p110γ in neutrophils from mice lacking both the p101 and p84 regulatory subunits, indicating that RAS binding to p110γ is insufficient to support GPCR activation in this cell type. These findings define a direct role for Gβγ subunits in activating both of the endogenous PI3Kγ complexes and indicate that the regulatory PI3Kγ subunit biases activation toward different GPCRs., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Limited information is available regarding male reproductive physiology in West Indian manatees ( Trichechus manatus ). Currently, only basic ultrastructural and morphometric descriptions of the spermatozoon exist; however, there are no reports evaluating any seminal characteristics in this species. Therefore, the aim of the study was to fill current gaps in knowledge regarding semen parameters in West Indian manatees by collecting and characterizing multiple ejaculate samples from a single, adult West Indian manatee. Samples were analyzed for the following semen parameters: volume, agglutination, pH, osmolality, viscosity, concentration, total sperm number, motility and kinematic parameters, morphology, plasma membrane integrity, acrosome integrity, chromatin maturation, and chromatin condensation. All macroscopic semen parameters varied to some extent between samples. Total and progressive motility was high for ejaculates 2 to 5, exceeding 97 and 89%, respectively; however, these parameters decreased dramatically throughout ejaculates 6 and 7. Across all samples, curvilinear velocity, straight-line velocity, and average pathway velocity represented the largest significant differences ( p < 0.001) between each of the progression velocity subgroups (rapid, medium, slow). Sperm characteristics, including acrosome integrity (79.8%), chromatin condensation (93.1%), and chromatin maturation (99.5%) were very high; however, high numbers of morphologically abnormal sperm were present (52.9%) and plasma membrane integrity was low (45.1%). These results are the first of their kind for this species and suggest high semen quality, based on multiple ejaculates, in this male West Indian manatee., (Copyright © 2020 Cowart, Collins, Mignucci-Giannoni, Alejandro-Zayas, Rivera-Guzman and Larkin.)

Introduction Triple negative breast cancer (TNBC) represents a heterogeneous subtype of breast cancer that carries a poorer prognosis. There remains a need to identify novel drivers of TNBC, which may represent targets to treat the disease. c-Met overexpression is linked with decreased survival and is associated with the basal subtype of breast cancer. Cpd A, a kinase inhibitor selective/specific for Met kinase has demonstrated preclinical anti-cancer efficacy in TNBC. We aimed to assess the anti-cancer efficacy of Cpd A when combined with Src kinase, ErbB-family or hepatocyte growth factor (HGF) inhibitors in TNBC cell lines. Methods We determined the anti-proliferative effects of Cpd A, rilotumumab, neratinib and saracatinib tested alone and in combination in a panel of TNBC cells by acid phosphatase assays. We performed reverse phase protein array analysis of c-Met and IGF1Rβ expression and phosphorylation of c-Met (Y1234/1235) in TNBC cells and correlated their expression/phosphorylation with Cpd A sensitivity. We examined the impact of Cpd A, neratinib and saracatinib tested alone and in combination on invasive potential and colony formation.Results TNBC cells are not inherently sensitive to Cpd A, and neither c-Met expression nor phosphorylation are biomarkers of sensitivity to Cpd A. Cpd A enhanced the anti-proliferative effects of neratinib in vitro; however, this effect was limited to cell lines with innate sensitivity to Cpd A. Cpd A had limited anti-invasive effects but it reduced colony formation in the TNBC cell line panel.Conclusions Despite Cpd A having a potential role in reducing cancer cell metastasis, identification of strong predictive biomarkers of c-Met sensitivity would be essential to the development of a c-Met targeted treatment for an appropriately selected cohort of TNBC patients.

Purpose: Information Communication Technology, such as smartphones, apps, the internet, etc., has become all-pervasive in our society. To learn the impact of Information Communication Technology (ICT) on everyday functioning, specifically Instrumental Activities of Daily Living (IADL), an exploratory review was undertaken. Methods: We identified how many primary IADL domains were substantially influenced by ICT, by examining primary IADL domains, in terms of traditional and ICT-enabled ways of completing these common tasks. The study further explored the implications of the pervasiveness of ICT and the ICT-enabled new normal in everyday functioning that underpins the need to redefine IADLs. Results: This examination revealed that the impact of ICT on IADLs has been so profound, that it marks a paradigm shift in the way we assess IADL completion. Creation of the concept of "Electronic Instrumental Activities of Daily Living (eIADLs)" would be a first step in marking this paradigm shift, and would foster further advances for understanding the mechanisms underlying the ICT-based life style related outcome measurement in rehabilitation science.IMPLICATIONS FOR REHABILITATIONThe presence of Information Communication Technology (ICT) has substantially influenced the way we complete our Instrumental Activities of Daily Living (IADL). Indeed, the impact is so profound, that it marks a paradigm shift in the way we should assess and measure everyday functioning.Creation of the concept of eIADLs would be a first step in marking this paradigm shift, and would foster further advances for understanding the mechanisms underlying the ICT-based life style related outcome measurement in the rehabilitation sciences.

Background: Drug overdose is the leading cause of death after release from prison, and this risk is significantly higher among women compared to men. Within the first 2 weeks after release, the risk of death from drug overdose is 12.7 times higher than the general population, with risk of death further elevated among females. Although female inmates have higher rates of opioid use disorder and post-release overdose fatality, justice-involved women are under-represented in studies of medications for opioid use disorder. The Reducing Overdose After Release from Incarceration (ROAR) pilot intervention and evaluation (recruitment June 2019 through December 2020) aims to reduce opioid overdose among women released to the community following incarceration in state prison. The evaluation further assesses induction, acceptance and effectiveness of extended release naltrexone in a female post-prison population., Methods/design: In the week prior to their release, female adults in custody with moderate to severe opioid use disorder start treatment with extended release naltrexone, an injectable opioid antagonist that blocks the effects of opioids for up to 1 month. All ROAR participants receive training to use naloxone rescue kits and are provided nasal naloxone at release. Ongoing support from a certified recovery mentor to facilitate sustained engagement with treatment for substance use disorders begins in the month prior to release from prison and continues for 6 months in community. We evaluate the association between ROAR participation and the primary outcome of opioid overdose. Using administrative data provided by the Oregon Department of Corrections and the Oregon Health Authority, we compare the odds of overdose among ROAR participants versus a comparison group of females released from prison during the study period. Evaluation activities in community includes survey and qualitative interviews for 6 months post release, as well as a review of clinic records to assess retention on medication among the pilot cohort (N = 100)., Discussion: ROAR is a collaboration between Oregon's public health, criminal justice, and medical communities. The ROAR intervention and evaluation provide critical information on improving interventions to prevent opioid overdose and improve retention on treatment in community in an overlooked, high-risk population: incarcerated women re-entering the community., Trial Registration: Clinical Trials.gov TRN: NCT03902821 .

Electron backscatter diffraction (EBSD) is a well-established method of characterisation for crystalline materials. Using this technique, we can rapidly acquire and index diffraction patterns to provide phase and orientation information about the crystals on the material surface. The conventional analysis method uses signal processing based on a Hough/Radon transform to index each diffraction pattern. This method is limited to the analysis of simple geometric features and ignores subtle characteristics of diffraction patterns, such as variations in relative band intensities. A second method, developed to address the shortcomings of the Hough/Radon transform, is based on template matching of a test experimental pattern with a large library of potential patterns. In the present work, the template matching approach has been refined with a new cross correlation function that allows for a smaller library and enables a dramatic speed up in pattern indexing. Refinement of the indexed orientation is performed with a follow-up step to allow for small alterations to the best match from the library search. The refined template matching approach is shown to be comparable in accuracy, precision and sensitivity to the Hough based method, even exceeding it in some cases, via the use of simulations and experimental data collected from a silicon single crystal and a deformed α-iron sample. The speed up and pattern refinement approaches should increase the widespread utility of pattern matching approaches., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Despite trastuzumab and pertuzumab improving outcome for patients with HER2-positive metastatic breast cancer, the disease remains fatal for the majority of patients. This study evaluated the anti-proliferative effects of adding anti-HER2 tyrosine kinase inhibitors (TKIs) to trastuzumab and pertuzumab in HER2-positive breast cancer cells. Afatinib was tested alone and in combination with trastuzumab in HER2-positive breast cancer cell lines. TKIs (lapatinib, neratinib, afatinib) combined with trastuzumab and/or pertuzumab were tested in 3 cell lines, with/without amphiregulin and heregulin-1β. Seven of 11 HER2-positive cell lines tested were sensitive to afatinib (IC 50  < 80 nM). Afatinib plus trastuzumab produced synergistic growth inhibition in eight cell lines. In trastuzumab-sensitive SKBR3 cells, the TKIs enhanced response to trastuzumab. Pertuzumab alone did not inhibit growth and did not enhance trastuzumab-induced growth inhibition or antibody-dependent cellular cytotoxicity. Pertuzumab enhanced response to trastuzumab when combined with lapatinib but not neratinib or afatinib. In two trastuzumab-resistant cell lines, the TKIs inhibited growth but adding trastuzumab and/or pertuzumab did not improve response compared to TKIs alone. Amphiregulin plus heregulin-1β stimulated proliferation of SKBR3 and MDA-MB-453 cells. In the presence of the growth factors, neither antibody inhibited growth and the TKIs showed significantly reduced activity. The triple combination of trastuzumab, pertuzumab and a TKI showed the strongest anti-proliferative activity in all three cell lines, in the presence of exogenous growth factors. In summary, addition of anti-HER2 TKIs to combined anti-HER2 monoclonal antibody therapy results in enhanced anticancer activity. These data contribute to the rationale for studying maximum HER2 blockade in the clinic.

Flourishing is an indicator of positive mental health and is important for children's development and well-being. We used variables from the National Survey of Children's Health 2016 as indicators of flourishing (difficulty making friends, is bullied, bullies others, shares ideas with family, argues, finishes tasks, does all homework, shows curiosity, stays calm, and cares about doing well in school) to compare differences in parent perceptions of their children with and without autism spectrum disorder (ASD). We anticipate that these findings will help identify intervention targets to support the well-being of individuals with ASD. Children between 6 and 17 years of age, without intellectual disability, brain injury, cerebral palsy, or Down syndrome were included. Total participants were 34,171 controls (male/female = 17,116/17,155) and 812 with ASD (male/female = 668/144). Factor analysis resulted in three-factor structures (social competence, behavioral control, and school motivation) with good model fit (root mean square error of approximation = 0.08, comparative fit index = 0.92, Tucker-Lewis index = 0.89). The multivariate regression model and propensity score with inverse probability of treatment weighting (PS-IPTW) method revealed that children with ASD had lower scores in the social competence and behavioral control factors compared to the control group (all P < 0.05). However, no significant differences were found in the school motivation factor between the two groups (P > 0.05) in both multivariate regression model and PS-IPTW method. Findings suggest that social competence and behavioral control are indicators of flourishing and are important intervention targets to increase flourishing among children with ASD. Autism Res 2019, 12: 952-966. © 2019 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: Flourishing is an indicator of positive mental health and is important for children's development and well-being. We used variables from The National Survey of Children's Health 2016 to examine differences in parent perceptions of the indicators of flourishing (difficulty making friends, is bullied, bullies others, shares ideas with family, argues, finishes tasks, does all homework, shows curiosity, stays calm, and cares about doing well in school) between children with and without autism spectrum disorders (ASD). We anticipate that this information will help to identify therapeutic targets to support the well-being of individuals with ASD. Children between 6 and 17 years old, without intellectual disability (ID), brain injury (BI), cerebral palsy (CP), or Down syndrome (DS) were included. From the total (N = 50,212), we excluded children under age 6 (n = 14,494), those who once, but do not currently have ASD (n = 81), and those with ID (n = 432), BI (n = 170), CP (n = 35), and DS (n = 17), resulting in 34,983 records used. Total participants, age 6-17 years, were 34,171 controls (male/female = 17,116/17,155) and 812 with ASD (male/female = 668/144). Factor analysis resulted in the identification of three flourishing categories among the indicator variables (social competence, behavioral control, and school motivation). Children with ASD had lower scores in the social competence and behavioral control factors compared to the control group. However, there were no significant differences in the school motivation factor between the two groups. Findings suggest that social competence and behavioral control are indicators of flourishing and are important intervention targets to increase flourishing among children with ASD., (© 2019 International Society for Autism Research, Wiley Periodicals, Inc.)

An estimated 15-20% of breast cancers overexpress human epidermal growth factor receptor 2 (HER2/ERBB2/neu). Two small-molecule tyrosine kinase inhibitors (TKIs), lapatinib and neratinib, have been approved for the treatment of HER2-positive (HER2+) breast cancer. Lapatinib, a reversible epidermal growth factor receptor (EGFR/ERBB1/HER1) and HER2 TKI, is used for the treatment of advanced HER2+ breast cancer in combination with capecitabine, in combination with trastuzumab in patients with hormone receptor-negative metastatic breast cancer, and in combination with an aromatase inhibitor for the first-line treatment of HER2+ breast cancer. Neratinib, a next-generation, irreversible pan-HER TKI, is used in the US for extended adjuvant treatment of adult patients with early-stage HER2+ breast cancer following 1 year of trastuzumab. In Europe, neratinib is used in the extended adjuvant treatment of adult patients with early-stage hormone receptor-positive HER2+ breast cancer who are less than 1 year from the completion of prior adjuvant trastuzumab-based therapy. Preclinical studies have shown that these agents have distinct properties that may impact their clinical activity. This review describes the preclinical characterization of lapatinib and neratinib, with a focus on the differences between these two agents that may have implications for patient management., Competing Interests: C.S.V. and S.K. are the founders of Sinopsee Therapeutics, a biotechnology company developing molecules for therapeutic purposes; the current work has no conflict with the company. D.M.C., N.T.C. and J.C. have received research funding from Puma Biotechnology, Inc. D.M.C. has received consultancy fees from Puma Biotechnology, Inc. J.C. has participated in advisory boards for Puma Biotechnology, Inc. and Novartis Oncology. A.S.L. and L.D.E. are employees and shareholders of Puma Biotechnology, Inc.

Antibody-drug conjugates (ADCs) combine the tumor selectivity of antibodies with the potency of cytotoxic small molecules thereby constituting antibody-mediated chemotherapy. As this inherently limits the adverse effects of the chemotherapeutic, such approaches are heavily pursued by pharma and biotech companies and have resulted in four FDA (Food and Drug Administration)-approved ADCs. However, as with other cancer therapies, durable responses are limited by the fact that under cell stress exerted by these drugs, tumors can acquire mechanisms of escape. Resistance can develop against the antibody component of ADCs by down-regulation/mutation of the targeted cell surface antigen or against payload toxicity by up-regulation of drug efflux transporters. Unique resistance mechanisms specific for the mode of action of ADCs have also emerged, like altered internalization or cell surface recycling of the targeted tumor antigen, changes in the intracellular routing or processing of ADCs, and impaired release of the toxic payload into the cytosol. These evasive changes are tailored to the specific nature and interplay of the three ADC constituents: the antibody, the linker, and the payload. Hence, they do not necessarily endow broad resistance to ADC therapy. This review summarizes preclinical and clinical findings that shed light on the mechanisms of acquired resistance to ADC therapies.

Mycobacterium pinnipedii causes tuberculosis in a number of pinniped species, and transmission to cattle and humans has been reported. The aims of this study were to: characterize the pathology and prevalence of tuberculosis in New Zealand marine mammals; use molecular diagnostic methods to confirm and type the causal agent; and to explore relationships between type and host characteristics. Tuberculosis was diagnosed in 30 pinnipeds and one cetacean. Most affected pinnipeds had involvement of the pulmonary system, supporting inhalation as the most common route of infection, although ingestion was a possible route in the cetacean. PCR for the RD2 gene confirmed M. pinnipedii as the causal agent in 23/31 (74%) cases (22 using DNA from cultured organisms, and one using DNA from formalin-fixed paraffin-embedded (FFPE) tissue), including the first published report in a cetacean. RD2 PCR results were compared for 22 cases where both cultured organisms and FFPE tissues were available, with successful identification of M. pinnipedii in 7/22 (31.8%). In cases with moderate to large numbers of acid-fast bacilli, RD2 PCR on FFPE tissue provided a rapid, inexpensive method for confirming M. pinnipedii infection without the need for culture. VNTR typing distinguished New Zealand M. pinnipedii isolates from M. pinnipedii isolated from Australian pinnipeds and from common types of M. bovis in New Zealand. Most (16/18) M. pinnipedii isolates from New Zealand sea lions were one of two common VNTR types whereas the cetacean isolate was a type detected previously in New Zealand cattle., Competing Interests: The authors have declared that no competing interests exist.

Multivariate statistical methods are widely used throughout the sciences, including microscopy, however, their utilisation for analysis of electron backscatter diffraction (EBSD) data has not been adequately explored. The basic aim of most EBSD analysis is to segment the spatial domain to reveal and quantify the microstructure, and links this to knowledge of the crystallography (e.g. crystal phase, orientation) within each segmented region. Two analysis strategies have been explored; principal component analysis (PCA) and k-means clustering. The intensity at individual (binned) pixels on the detector were used as the variables defining the multidimensional space in which each pattern in the map generates a single discrete point. PCA analysis alone did not work well but rotating factors to the VARIMAX solution did. K-means clustering also successfully segmented the data but was computational more expensive. The characteristic patterns produced by either VARIMAX or k-means clustering enhance weak patterns, remove pattern overlap, and allow subtle effects from polarity to be distinguished. Combining multivariate statistical analysis (MSA) approaches with template matching to simulation libraries can significantly reduce computational demand as the number of patterns to be matched is drastically reduced. Both template matching and MSA approaches may augment existing analysis methods but will not replace them in the majority of applications., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The ability to DNA fingerprint Mycobacterium bovis isolates helped to define the role of wildlife in the persistence of bovine tuberculosis in New Zealand. DNA fingerprinting results currently help to guide wildlife control measures and also aid in tracing the source of infections that result from movement of livestock. During the last 5 years we have developed the ability to distinguish New Zealand (NZ) M. bovis isolates by comparing the sequences of whole genome sequenced (WGS) M. bovis samples. WGS provides much higher resolution than our other established typing methods and greatly improves the definition of the regional localization of NZ M. bovis types. Three outbreak investigations are described and results demonstrate how WGS analysis has led to the confirmation of epidemiological sourcing of infection, to better definition of new sources of infection by ruling out other possible sources, and has revealed probable wildlife infection in an area considered to be free of infected wildlife. The routine use of WGS analyses for sourcing new M. bovis infections will be an important component of the strategy employed to eradicate bovine TB from NZ livestock and wildlife.

As HER2 is a client protein of the molecular chaperone Hsp90, targeting Hsp90 may be beneficial in HER2-positive breast cancer. In this study, the activity of the Hsp90 inhibitor NVP-AUY922 was assessed in HER2 overexpressing breast cancer cell lines, including two cell line models of acquired trastuzumab-resistance. The seven HER2-positive breast cancer cell lines tested showed significant sensitivity to NVP-AUY922 in vitro, with IC 50 values between 6 and 17 nM. Combining NVP-AUY922 with chemotherapy did not improve response. NVP-AUY922 in combination with trastuzumab, significantly enhanced growth inhibition in three of the seven cell lines tested. In conclusion, our data shows that NVP-AUY922 displays potent anti-cancer activity in HER2-positive and trastuzumab-resistant breast cancer cells, and supports further testing of NVP-AUY922 in patients with HER2-positive breast cancer.