Addressing Gender-Based Violence Using Evidence-Based Practices During COVID-19: The Case of Puerto Rico.

As gender-based violence (GBV) surged during the COVID-19 pandemic, the 65th session of the Commission on the Status of Women (CSW65) called for member states, civil, and other stakeholders to consider the specific needs of women and girls in COVID-19 response and recovery efforts. Psychology provides scientific knowledge to help answer this call. Despite existing global guidance and psychological research to mitigate GBV, COVID-19 presents new challenges for consideration. This article summarizes existing GBV guidance/research and COVID-19 considerations, uses an illustrative case study to describe Puerto Rico's application of GBV guidance/research during COVID-19, and provides preliminary policy and practice recommendations.

The flip side of traditional nursing education: A literature review.

The flipped classroom (FC) andragogy purports an improvement of critical thinking and problem-solving skills in students. This literature review explores fourteen research studies and discusses outcome measures reported on the effectiveness of using this teaching modality. Students described the learning activities during the classroom meeting times as valuable and indicated the interaction and engagement were beneficial to their learning. Many students opined an increased comprehension of the subject matter. Overall, the FC required more work on the part of the students and the faculty, and the majority of students preferred the traditional classroom (TC) passive method of learning over the FC active learning andragogy as a result of the substantial time commitment required for preparation necessitated by the FC. Five of the fourteen studies evaluated student learning outcome measures; four studies showed an improvement in the FC environment compared to the TC and one reported the FC was at least as effective as the TC. Further studies with quantifiable outcome measures are required to determine the effectiveness of a FC on critical thinking and problem-solving skills of nursing students.

Self-Renewal and CSCs In Vitro Enrichment: Growth as Floating Spheres.

Cancer stem cells (CSC) are a vital component to the progression and reoccurrence of cancers, making them a primary target of study for both fundamental understanding of cancer biology and the development of effective and targeted treatments. CSCs reside in a complex 3D microenvironment, and the 3D spheroids are an indispensable tool in tumor biology due to their 3D structure and replication of the tumor microenvironment. Within this chapter the methodology for CSC isolation, suspension culture in hanging drop model, and characterization assays for CSC are described. First, the methodology for identifying and isolating CSCs from patient tumors, ascites, or cancer cell lines is described through the use of FACS analysis. Next, a detailed description of 3D hanging drop model for generating CSC spheroids is provided, followed by maintenance and monitoring techniques for extended 3D culture. Analysis methods are described for the quantification of CSC spheroid proliferation and viability tracking, throughout culture by on-plate alamarBlue fluorescence. Additional viability assays are described utilizing confocal microscopy with Live/Dead Viability/Cytotoxicity Kit. The characterization of CSCs populations within spheroids is described through FACS analysis. Further, an immunohistochemistry procedure is described for cell-cell and cell-matrix interaction assessment. Finally, several notes and tips for successful experiments with 3D CSC spheroids on the hanging drop model are provided. These methods are not only applicable to CSCs within a variety of tumor cell types, for not only understanding the fundamental tumor biology, but also for drug screening and development of preclinical chemotherapeutic strategies.

Personalized Medicine-Based Approach to Model Patterns of Chemoresistance and Tumor Recurrence Using Ovarian Cancer Stem Cell Spheroids.

Purpose: Chemoresistant ovarian cancers grow in suspension within the ascites fluid. To screen the effect of chemotherapeutics and biologics on resistant ovarian cancers with a personalized basis, we developed a 3D hanging drop spheroid platform.Experimental Design: We initiated spheroids with primary aldehyde dehydrogenase-positive (ALDH+) CD133+ ovarian cancer stem cells (OvCSC) from different patient samples and demonstrated that stem cell progeny from harvested spheroids was similar to the primary tumor. OvCSC spheroids were utilized to initiate tumors in immunodeficient mice. Drug responses to cisplatin and ALDH-targeting compound or JAK2 inhibitor determined whether the OvCSC population within the spheroids could be targeted. Cells that escaped therapy were isolated and used to initiate new spheroids and model tumor reemergence in a personalized manner.Results: OvCSC spheroids from different patients exhibited varying and personalized responses to chemotherapeutics. Xenografts were established from OvCSC spheroids, even with a single spheroid. Distinct responses to therapy were observed in distinct primary tumor xenografts similar to those observed in spheroids. Spheroids resistant to cisplatin/ALDH inhibitor therapy had persistent, albeit lower ALDH expression and complete loss of CD133 expression, whereas those resistant to cisplatin/JAK2 inhibitor therapy were enriched for ALDH+ cells.Conclusions: Our 3D hanging drop suspension platform can be used to propagate primary OvCSCs that represent individual patient tumors effectively by differentiating in vitro and initiating tumors in mice. Therefore, our platform can be used to study cancer stem cell biology and model tumor reemergence to identify new targeted therapeutics from an effective personalized medicine standpoint. Clin Cancer Res; 23(22); 6934-45. ©2017 AACR.

Comparative analysis of tumor spheroid generation techniques for differential in vitro drug toxicity.

Multicellular tumor spheroids are powerful in vitro models to perform preclinical chemosensitivity assays. We compare different methodologies to generate tumor spheroids in terms of resultant spheroid morphology, cellular arrangement and chemosensitivity. We used two cancer cell lines (MCF7 and OVCAR8) to generate spheroids using i) hanging drop array plates; ii) liquid overlay on ultra-low attachment plates; iii) liquid overlay on ultra-low attachment plates with rotating mixing (nutator plates). Analysis of spheroid morphometry indicated that cellular compaction was increased in spheroids generated on nutator and hanging drop array plates. Collagen staining also indicated higher compaction and remodeling in tumor spheroids on nutator and hanging drop arrays compared to conventional liquid overlay. Consequently, spheroids generated on nutator or hanging drop plates had increased chemoresistance to cisplatin treatment (20-60% viability) compared to spheroids on ultra low attachment plates (10-20% viability). Lastly, we used a mathematical model to demonstrate minimal changes in oxygen and cisplatin diffusion within experimentally generated spheroids. Our results demonstrate that in vitro methods of tumor spheroid generation result in varied cellular arrangement and chemosensitivity.

Formation of stable small cell number three-dimensional ovarian cancer spheroids using hanging drop arrays for preclinical drug sensitivity assays.

BACKGROUND: Ovarian cancer grows and metastasizes from multicellular spheroidal aggregates within the ascites fluid. Multicellular tumor spheroids are therefore physiologically significant 3D in vitro models for ovarian cancer research. Conventional hanging drop cultures require high starting cell numbers, and are tedious for long-term maintenance. In this study, we generate stable, uniform multicellular spheroids using very small number of ovarian cancer cells in a novel 384 well hanging drop array platform. METHODS: We used novel tumor spheroid platform and two ovarian cancer cell lines (A2780 and OVCAR3) to demonstrate the stable incorporation of as few as 10 cells into a single spheroid. RESULTS: Spheroids had uniform geometry, with projected areas (42.60×10(3)µm-475.22×10(3)µm(2) for A2780 spheroids and 37.24×10(3)µm(2)-281.01×10(3)µm(2) for OVCAR3 spheroids) that varied as a function of the initial cell seeding density. Phalloidin and nuclear stains indicated cells formed tightly packed spheroids with demarcated boundaries and cell-cell interaction within spheroids. Cells within spheroids demonstrated over 85% viability. 3D tumor spheroids demonstrated greater resistance (70-80% viability) to cisplatin chemotherapy compared to 2D cultures (30-50% viability). CONCLUSIONS: Ovarian cancer spheroids can be generated from limited cell numbers in high throughput 384 well plates with high viability. Spheroids demonstrate therapeutic resistance relative to cells in traditional 2D culture. Stable incorporation of low cell numbers is advantageous when translating this research to rare patient-derived cells. This system can be used to understand ovarian cancer spheroid biology, as well as carry out preclinical drug sensitivity assays.

Working patterns and vitamin D status in mid-life: a cross-sectional study of the 1958 British birth cohort.

OBJECTIVES: To assess the association between working patterns and vitamin D status in men and women and to determine the potential influence of related lifestyle and socioeconomic factors. METHODS: The authors used data from the 1958 British birth cohort (aged 45 years) and 6154 participants, who were in full-time work, were included in current analyses. Vitamin D status was measured by circulating concentrations of 25-hydroxyvitamin D (25(OH)D). Information on working patterns and lifestyle factors was obtained using a structured questionnaire administered at 45 years. RESULTS: Manual social class was strongly associated with vitamin D-related lifestyle factors, with those in manual classes not only spending more time outdoors, but also spending more time watching TV/using PC, consuming less supplements and oily fish. Associations between working patterns and vitamin D-related lifestyles were less clear: night work was not strongly associated with lifestyles in either gender, while working hours were associated with time spent outside, PC/TV leisure time and use of supplements in men but not in women. In men, working patterns were not associated with lower 25(OH)D concentrations. In women, 25(OH)D concentrations were 8% lower (95% CI 15% to 2%) in night workers compared with others, while women working less than 35 h/week had 5% higher concentrations of 25(OH)D (95% CI 1% to 8%) compared with those working 35-40 h/week after adjustment for season, social class and body mass index (BMI). CONCLUSIONS: Women working nights and longer hours may be vulnerable to deficits in vitamin D status and associated health hazards.

Similar age-specific PSA, complexed PSA, and percent cPSA levels among African-American and white men of southern Louisiana. Prostate-specific antigen.

OBJECTIVES: To determine the mean and median complexed prostate-specific antigen (cPSA) levels in a predominantly African-American population and to explore whether differences in cPSA exist between the races. Differences in total PSA (tPSA) levels between age-matched African-American and white men have been reported by several groups. Age-specific cPSA levels, however, have not been thoroughly evaluated and reported among African-American populations. METHODS: We prospectively evaluated the serum cPSA, tPSA, and percent cPSA levels by the Bayer Immuno 1 assay as a function of age among 1755 African-American and 630 white men in southern Louisiana presenting to a prostate cancer screening program. All men had a normal digital rectal examination or biopsy-proven benign pathologic findings. The intragroup and intergroup statistical analyses were carried out for each decade of age. RESULTS: The median cPSA level for African-American men aged 40 to 49, 50 to 59, and 60 to 69 years old was 0.45, 0.61, and 0.84 ng/mL, respectively. These did not differ significantly from those of age-matched white men. Also, no difference was found in the mean or median tPSA between the races. The percent cPSA values showed a trend for lower levels in African Americans. CONCLUSIONS: These findings suggest that no significant difference exists between cPSA or tPSA levels in African-American and white men without prostate cancer. Genetic and epigenetic factors distinct to this region may account for this observation and thus need further evaluation.