Transthoracic ultrasound-guided biopsy in the hands of chest physicians – a stepwise approach

Background: The evaluation of patients with lung lesions is challenging. The nature of the lesion can be determined by pathological evaluation of biopsies. The pulmonologists will be met by increasing demands with regard to biopsy techniques including ultrasound-guided transthoracic needle biopsy (US-TTNB).Objective: The aim of this paper is to present the pulmonologist to a systematic step-by-step guide for performing US-TTNB and to assess the evidence for this approach. Method/results: Indications, contraindications and a step-by-step guide for the techniques used when performing US-TTNB are presented, and major complications and handling of these are described. Conclusion: US-TTNB performed by pulmonologists is a safe and feasible procedure

CETSA-based target engagement of taxanes as biomarkers for efficacy and resistance

The use of taxanes has for decades been crucial for treatment of several cancers. A major limitation of these therapies is inherent or acquired drug resistance. A key to improved outcome of taxane-based therapies is to develop tools to predict and monitor drug efficacy and resistance in the clinical setting allowing for treatment and dose stratification for individual patients. To assess treatment efficacy up to the level of drug target engagement, we have established several formats of tubulin-specific Cellular Thermal Shift Assays (CETSAs). This technique was evaluated in breast and prostate cancer models and in a cohort of breast cancer patients. Here we show that taxanes induce significant CETSA shifts in cell lines as well as in animal models including patient-derived xenograft (PDX) models. Furthermore, isothermal dose response CETSA measurements allowed for drugs to be rapidly ranked according to their reported potency. Using multidrug resistant cancer cell lines and taxane-resistant PDX models we demonstrate that CETSA can identify taxane resistance up to the level of target engagement. An imaging-based CETSA format was also established, which in principle allows for taxane target engagement to be accessed in specific cell types in complex cell mixtures. Using a highly sensitive implementation of CETSA, we measured target engagement in fine needle aspirates from breast cancer patients, revealing a range of different sensitivities. Together, our data support that CETSA is a robust tool for assessing taxane target engagement in preclinical models and clinical material and therefore should be evaluated as a prognostic tool during taxane-based therapies

Tumor-specific HMG-CoA reductase expression in primary premenopausal breast cancer predicts response to tamoxifen

ABSTRACT: INTRODUCTION: We previously reported an association between tumor-specific 3-hydroxy-3-methylglutharyl-coenzyme A reductase (HMG-CoAR) expression and a good prognosis in breast cancer. Here, the predictive value of HMG-CoAR expression in relation to tamoxifen response was examined. METHODS: HMG-CoAR protein and RNA expression was analyzed in a cell line model of tamoxifen resistance using western blotting and PCR. HMG-CoAR mRNA expression was examined in 155 tamoxifen-treated breast tumors obtained from a previously published gene expression study (Cohort I). HMG-CoAR protein expression was examined in 422 stage II premenopausal breast cancer patients, who had previously participated in a randomized control trial comparing 2 years of tamoxifen with no systemic adjuvant treatment (Cohort II). Kaplan-Meier analysis and Cox proportional hazards modeling were used to estimate the risk of recurrence-free survival (RFS) and the effect of HMG-CoAR expression on tamoxifen response. RESULTS: HMG-CoAR protein and RNA expression were decreased in tamoxifen-resistant MCF7-LCC9 cells compared with their tamoxifen-sensitive parental cell line. HMG-CoAR mRNA expression was decreased in tumors that recurred following tamoxifen treatment (P < 0.001) and was an independent predictor of RFS in Cohort I (hazard ratio = 0.63, P = 0.009). In Cohort II, adjuvant tamoxifen increased RFS in HMG-CoAR-positive tumors (P = 0.008). Multivariate Cox regression analysis demonstrated that HMG-CoAR was an independent predictor of improved RFS in Cohort II (hazard ratio = 0.67, P = 0.010), and subset analysis revealed that this was maintained in estrogen receptor (ER)-positive patients (hazard ratio = 0.65, P = 0.029). Multivariate interaction analysis demonstrated a difference in tamoxifen efficacy relative to HMG-CoAR expression (P = 0.05). Analysis of tamoxifen response revealed that patients with ER-positive/HMG-CoAR tumors had a significant response to tamoxifen (P = 0.010) as well as patients with ER-positive or HMG-CoAR-positive tumors (P = 0.035). Stratification according to ER and HMG-CoAR status demonstrated that ER-positive/HMG-CoAR-positive tumors had an improved RFS compared with ER-positive/HMG-CoAR-negative tumors in the treatment arm (P = 0.033); this effect was lost in the control arm (P = 0.138), however, suggesting that HMG-CoAR predicts tamoxifen response. CONCLUSIONS: HMG-CoAR expression is a predictor of response to tamoxifen in both ER-positive and ER-negative disease. Premenopausal patients with tumors that express ER or HMG-CoAR respond to adjuvant tamoxifen

Structure and mechanism of Zn^(2+)- transporting P-type ATPases

Zinc is an essential micronutrient for all living organisms. It is required for signalling and proper functioning of a range of proteins involved in, for example, DNA binding and enzymatic catalysis. In prokaryotes and photosynthetic eukaryotes, Zn2+-transporting P-type ATPases of class IB (ZntA) are crucial for cellular redistribution and detoxification of Zn2+ and related elements. Here we present crystal structures representing the phosphoenzyme ground state (E2P) and a dephosphorylation intermediate (E2·P_i) of ZntA from Shigella sonnei, determined at 3.2 Å and 2.7 Å resolution, respectively. The structures reveal a similar fold to Cu^+-ATPases, with an amphipathic helix at the membrane interface. A conserved electronegative funnel connects this region to the intramembranous high-affinity ion-binding site and may promote specific uptake of cellular Zn^(2+) ions by the transporter. The E2P structure displays a wide extracellular release pathway reaching the invariant residues at the high-affinity site, including C392, C394 and D714. The pathway closes in the E2·P_i state, in which D714 interacts with the conserved residue K693, which possibly stimulates Zn^(2+) release as a built-in counter ion, as has been proposed for H^+-ATPases. Indeed, transport studies in liposomes provide experimental support for ZntA activity without counter transport. These findings suggest a mechanistic link between P_(IB)-type Zn^(2+)-ATPases and P_(III)-type H^+-ATPases and at the same time show structural features of the extracellular release pathway that resemble P_(II)-type ATPases such as the sarcoplasmic/endoplasmic reticulum Ca^(2+)-ATPase (SERCA) and Na^+, K^+-ATPase. These findings considerably increase our understanding of zinc transport in cells and represent new possibilities for biotechnology and biomedicine

Simulering i sykepleierutdannelsen

Hensikt. Studiet belyser viktigheten av simulering med tanke på de ikke tekniske ferdighetene. Fokuset er rettet mot fem viktige temaer innen simulering; kommunikasjon, ikke-tekniske ferdigheter, tverrfaglig teamarbeid, pasientsikkerhet og refleksjon. Formålet var å se nytteeffekten av simulering i sykepleierutdannelsen og dens overføringsverdi til profesjon i kommende yrke. Metode. Litteraturstudie med kvalitativ metode. Det ble søkt i ulike databaser og åtte resultats artikler ble benyttet etter egne inklusjons- og eksklusjonskriterier. Artiklene ble analysert og vurdert etter gitte kriterier, samt kontrollert opp mot NSD (Norsk Senter for Forskningsdata). I tillegg benyttet vi teoretikere, relevant fagstoff og forskningspublikasjoner i studiet. Resultat. Benyttelse av simulering gir en positiv gevinst når det gjelder overgangen fra student til yrkesaktiv profesjon, men forskning kan ikke entydig dokumentere økt kunnskap i klinisk praksis. Kommunikasjon mellom profesjoner er essensielt i et tverrfaglig miljø, og bidrar til økt pasientsikkerhet. Bruk av kommunikasjonsverktøy, eksempelvis SBAR og læringsmetoder som Team STEPPS ved simulering viser forbedret tverrfaglig samarbeid, sikrere pasientbehandling og korrekt og målrettet kommunikasjon. Brifing og debrifing ved simulering i trygge tilrettelagte miljøer fremmer fagutvikling og økte kliniske evner, samt studentenes selvtillit. Tverrprofesjonell simulering bidrar blant annet til å redusere hierarkiet og gir en bedre innsikt i hverandres fagfelt. Konklusjon. Studien viser nytteverdi av simulering innen ikke-tekniske ferdigheter, som kommunikasjon, samarbeid og refleksjon, dette fremmer pasientsikkerhet. Det belyses også at studentene er avhengige av kompetente og oppdaterte veiledere, samt gode læringsmiljøer for å fremme god fagutvikling