Quantitation of total antibody (tAb) from antibody drug conjugate (ADC) PYX-201 in rat and monkey plasma using an enzyme-linked immunosorbent assay (ELISA) and its application in preclinical studies.

PYX-201 is an investigative ADC oncology drug composed of a monoclonal human immunoglobulin G (IgG) antibody targeting the extra domain B splice variant of fibronectin (EDB + FN) conjugated to an auristatin payload through a cleavable linker. Effective measurement of PYX-201 tAb is the key to ADC drug PYX-201 preclinical pharmacokinetics (PK) assessment. PYX-201 monoclonal antibody (mAb) was used as the reference standard, goat anti-human IgG polyclonal antibody (pAb) or rabbit anti-human Kappa light chain mAb was employed as the capture antibody, and mouse mAb or goat pAb anti-human IgG the crystallizable fragment (Fc) (horseradish peroxidase (HRP)) was utilized as the detection antibody in this ELISA. This assay was validated with a dynamic range 250 - 10,000 ng/mL and 250 - 6000 ng/mL in rat and monkey K2EDTA plasma, respectively. PYX-201 tAb bioanalytical ELISA assay was reported for the first time in any biological matrix. This is the first time for a bioanalytical method to be validated for a tAb from an ADC drug targeting EDB + FN in any biological matrix.

Quantification of antibody-drug conjugate PYX-201 in rat and monkey plasma via ELISA and its application in preclinical studies.

Aim: PYX-201 is a novel antibody-drug conjugate targeting the extra domain B splice variant of fibronectin in the tumor microenvironment. Accurate quantification of PYX-201 is critical for PYX-201 pharmacokinetics profiling in preclinical studies. Materials & methods: ELISA was performed using reference standard PYX-201, mouse monoclonal anti-monomethyl auristatin E antibody, mouse IgG1, mouse monoclonal anti-human IgG horseradish peroxidase and donkey anti-human IgG horseradish peroxidase. Results: This assay was validated at 50.0-10,000 ng/ml in rat dipotassium EDTA plasma and 250-10,000 ng/ml in monkey dipotassium EDTA plasma. Conclusion: This is the first time for a PYX-201 bioanalytical assay in any matrix to be reported.

Feasibility Using Amazon Mechanical Turk for Online Surveys of Attitudes and Perceptions of Chiropractic Health Care in the United States.

OBJECTIVE: The purpose of this study was to determine the feasibility and utility of using Amazon Mechanical Turk (MTurk) for online surveys of US adults about their perceptions of chiropractors and to report differences between people who have positive versus negative attitudes toward chiropractic care. METHODS: A 74-item, unvalidated survey was developed through iterative review to assess attitudes of respondents to chiropractic, including a query stratifying respondents based on previous and future use of chiropractic (have used, would consider; have used, would not consider; have not used, would consider; have not used, would not consider). The electronic survey was delivered using Qualtrics; respondents were recruited using MTurk, a crowdsourcing website. Descriptive statistics, including frequencies and cross tabulations, were performed. RESULTS: A total of 1300 responses were obtained. Consistent with previous reports, 32.2% of the respondents reported having seen a chiropractor in the past. Chiropractic care was perceived as being effective for musculoskeletal complaints. Respondents who would not consider future chiropractic care shared a common set of beliefs related to training of chiropractors, scope of chiropractic practice, and safety and reputation of chiropractic. These respondents reported increased likelihood of chiropractor use with the recommendation of a primary care physician. CONCLUSION: Recruiting survey participants using MTurk is feasible, affordable, and quick and offers high utility to academic researchers. Using this resource, we ascertained preliminary data about attitudes and perceptions from individuals who would or would not consider chiropractic, stratified by their previous use.

Modeling the effect of ascites-induced compression on ovarian cancer multicellular aggregates.

Epithelial ovarian cancer (EOC) is the most lethal gynecological malignancy. EOC dissemination is predominantly via direct extension of cells and multicellular aggregates (MCAs) into the peritoneal cavity, which adhere to and induce retraction of peritoneal mesothelium and proliferate in the submesothelial matrix to generate metastatic lesions. Metastasis is facilitated by the accumulation of malignant ascites (500 ml to >2 l), resulting in physical discomfort and abdominal distension, and leading to poor prognosis. Although intraperitoneal fluid pressure is normally subatmospheric, an average intraperitoneal pressure of 30 cmH2O (22.1 mmHg) has been reported in women with EOC. In this study, to enable experimental evaluation of the impact of high intraperitoneal pressure on EOC progression, two new in vitro model systems were developed. Initial experiments evaluated EOC MCAs in pressure vessels connected to an Instron to apply short-term compressive force. A Flexcell Compression Plus system was then used to enable longer-term compression of MCAs in custom-designed hydrogel carriers. Results show changes in the expression of genes related to epithelial-mesenchymal transition as well as altered dispersal of compressed MCAs on collagen gels. These new model systems have utility for future analyses of compression-induced mechanotransduction and the resulting impact on cellular responses related to intraperitoneal metastatic dissemination.This article has an associated First Person interview with the first authors of the paper.

The meaning of it all: evaluating knowledge of Minimal Clinically Important Difference (MCID) among chiropractic students.

INTRODUCTION: Patient-reported outcome measures are frequently used to monitor patient progress during chiropractic care, yet student interns utilizing such assessments are unfamiliar with what magnitude of change (MCID) is considered beneficial to the patient. OBJECTIVE: This work seeks to determine chiropractic intern knowledge of MCID. METHODS: A five-item survey was administered to 104 chiropractic student interns. RESULTS: Nearly one-third of the interns correctly defined the MCID acronym, and approximately one-third of the interns knew at least one MCID value for the outcome assessments in the EHR. Surprisingly, 20% of the interns reported knowledge of at least one MCID value, but answered incorrectly pertaining to the MCID acronym. CONCLUSION: Student interns value patient perception, but have limited knowledge of MCID values. Addressing this gap will improve their understanding of patient progress and inform their treatment decisions both in the outpatient clinic and in their practices following graduation.

INTRODUCTION: Les mesures de résultats rapportés par les patients sont fréquemment utilisées pour suivre les progrès du patient pendant les soins chiropratiques, mais les stagiaires qui utilisent ces évaluations ne sont pas familiers avec l'ampleur des changements (différence minimale cliniquement importante ­ DMCI) considérée comme bénéfique pour le patient. OBJECTIF: Cette étude vise à déterminer les connaissances, chez les stagiaires en chiropratique, de la DMCI. MÉTHODOLOGIE: Une enquête comportant cinq points a été menée chez 104 stagiaires en chiropratique. RÉSULTATS: Près d'un tiers des stagiaires ont correctement défini l'acronyme DMCI (MCID en anglais), et environ un tiers d'entre eux était au courant au moins d'une valeur de DMCI pour les évaluations de résultats dans le DSE. Étonnamment, 20 % des stagiaires ont indiqué connaître au moins une valeur de DMCI, mais n'ont pas su reconnaître l'acronyme DMCI. CONCLUSION: Les stagiaires accordent de l'importance à la perception des patients, mais ont une connaissance limitée des valeurs de DMCI. Combler cette lacune permettra d'améliorer leur compréhension des progrès des patients et d'informer leurs décisions de traitement tant en consultation externe que dans leurs pratiques après l'obtention du diplôme.

Wnt5a Signaling in Cancer.

Wnt5a is involved in activating several non-canonical WNT signaling pathways, through binding to different members of the Frizzled- and Ror-family receptors. Wnt5a signaling is critical for regulating normal developmental processes, including proliferation, differentiation, migration, adhesion and polarity. However, the aberrant activation or inhibition of Wnt5a signaling is emerging as an important event in cancer progression, exerting both oncogenic and tumor suppressive effects. Recent studies show the involvement of Wnt5a in regulating cancer cell invasion, metastasis, metabolism and inflammation. In this article, we review findings regarding the molecular mechanisms and roles of Wnt5a signaling in various cancer types, and highlight Wnt5a in ovarian cancer.