An Exploration of Doctor of Physical Therapy Students' Belongingness in Clinical Education: A Validation Study.

INTRODUCTION: The aim of this study was to adapt and validate the Belongingness Scale-Clinical Placement Experience (BES-CPE) for Doctor of Physical Therapy (DPT) students in the United States. REVIEW OF LITERATURE: Belongingness is vital to one's mental, emotional, and physical health. Research has shown that belongingness is positively correlated with students' academic performance and achievement. An absence of belongingness may hinder students' full participation in clinical experiences and compromise clinical achievement. SUBJECTS: Respondents were current or former DPT students at least 18 years of age who had either completed the midterm evaluation of their final terminal full-time clinical education experience (TCE) in their DPT program or were no more than 1 year from the completion of their final TCE. METHODS: The BES-CPE was adapted for DPT students, and the scale was completed electronically by those who met the inclusion criteria. Principal component analysis with promax rotation and Cronbach's α were used to determine construct validity and reliability. RESULTS: One hundred fifty-nine respondents completed all items on the BES-CPE and demographic survey. A 3-component structure was identified (esteem, connectedness, and efficacy), which was aligned to the original BES-CPE scale. One item was discarded, and the final version of the BES-CPE for DPT students is a 33-item scale with satisfactory internal consistency. DISCUSSION AND CONCLUSION: This study adapted and provided evidence for validity of the first known scale to measure belongingness in DPT students during their clinical education experiences (CEEs) in the United States. The 33-item BES-CPE provided valid and reliable measures of belongingness in DPT students during CEEs that can be used to provide a better understanding of the student experience in the clinical learning environment.

The post COVID-19 healthcare landscape and the use of long-acting injectable antipsychotics for individuals with schizophrenia and bipolar I disorder: the importance of an integrated collaborative-care approach.

BACKGROUND: Long-acting injectable antipsychotics (LAIs) are an essential maintenance treatment option for individuals with schizophrenia or bipolar I disorder (BP-I). This report summarizes a roundtable discussion on the impact of COVID-19 on the mental healthcare landscape and use of LAIs for individuals with schizophrenia or BP-I. METHODS: Ten experts and stakeholders from diverse fields of healthcare participated in a roundtable discussion on the impact of the COVID-19 pandemic, treatment challenges, and gaps in healthcare for individuals with schizophrenia or BP-I, informed by a literature search. RESULTS: Individuals with schizophrenia or BP-I are at increased risk of COVID-19 infection and increased risk of mortality after COVID-19 diagnosis. LAI prescriptions decreased early on in the pandemic, driven by a decrease in face-to-face consultations. Mental healthcare services are adapting with increased use of telehealth and home-based treatment. Clinical workflows to provide consistent, in-person LAI services include screening for COVID-19 exposure and infection, minimizing contact, and ensuring mask-wearing by individuals and staff. The importance of continued in-person visits for LAIs needs to be discussed so that staff can share that information with patients, their caregivers, and families. A fully integrated, collaborative-care model is the most important aspect of care for individuals with schizophrenia or BP-I during and after the COVID-19 pandemic. CONCLUSIONS: The COVID-19 pandemic has highlighted the importance of a fully integrated collaborative-care model to ensure regular, routine healthcare contact and access to prescribed treatments and services for individuals with schizophrenia and BP-I.

Sex-related differences in primary metastatic site in rectal cancer; associated with hemodynamic factors?

BACKGROUND AND PURPOSE: We investigated how features relating to pelvic cavity anatomy and tumor hemodynamic factors may influence systemic failure in rectal cancer. MATERIALS AND METHODS: Rectal cancer patients (207 women, 343 men), who had been prospectively enrolled onto six cohorts and given curative-intent therapy, were analyzed for the first metastatic event. In one of the cohorts, the diameter of the inferior mesenteric vein (IMV) was assessed on diagnostic abdominal computed tomography images (n = 113). Tumor volume (n = 193) and histologic response to neoadjuvant therapy (n = 445) were recorded from diagnostic magnetic resonance images and surgical specimens, respectively. RESULTS: More women than men developed lung metastasis (p = 0.037), while the opposite was the case for liver metastasis (p = 0.040). Wider IMV diameter correlated with larger tumor volume (r = 0.481, p < 0.001) and male sex (p < 0.001). Female sex was the only adverse prognostic factor for lung metastasis. When sex, tumor volume, and histologic response were taken into consideration, poor tumor response remained the only determinant for liver metastasis (p = 0.002). CONCLUSIONS: In a diverse rectal cancer population given curative-intent treatment, women and men had different outcome with regard to the primary metastatic site. Tumor hemodynamic factors should be considered in rectal cancer risk stratification.

Relationship of Omega-3 fatty acids DHA and EPA with the inflammatory biomarker hs-CRP in children with sickle cell anemia.

BACKGROUND: Inflammation and vaso-occlusion play key roles in Sickle Cell Disease (SCD) pathophysiology. Lipoxygenase products of the omega-3 fatty acids (O3FAs), docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids, are potent anti-inflammatory mediators modulating pain. O3FAs decrease episodes of vaso-occlusion in SCD. METHODS: We assessed erythrocyte fatty acid composition in two major cell membrane phospholipids, phosphatidylcholine and phosphatidylethanolamine, in children with SCD HbSS-disease (n = 38) and age/race-matched HbAA-controls (n = 18). Ratio of pro-inflammatory arachidonic acid (AA) to anti-inflammatory DHA and EPA (FA-Ratio), and its relationship to hs-CRP were evaluated. RESULTS: FA-Ratios were increased in both phosphatidylcholine and phosphatidylethanolamine in HbSS compared to controls. Correlations were noted in HbSS subjects between hs-CRP and FA-Ratios (p = 0.011). FA-Ratios increased with age (p = 0.0007) due to an increase in pro-inflammatory AA with a concomitant decrease in anti-inflammatory DHA. CONCLUSIONS: Findings demonstrate relative deficiencies in HbSS of the anti-inflammatory precursor fatty acids DHA and EPA, which correlates positively with hs-CRP.

Quantitative sensory testing in children with sickle cell disease: additional insights and future possibilities.

Quantitative sensory testing (QST) is used in a variety of pain disorders to characterize pain and predict prognosis and response to specific therapies. In this study, we aimed to confirm results in the literature documenting altered QST thresholds in sickle cell disease (SCD) and assess the test-retest reliability of results over time. Fifty-seven SCD and 60 control subjects aged 8-20 years underwent heat and cold detection and pain threshold testing using a Medoc TSAII. Participants were tested at baseline and 3 months; SCD subjects were additionally tested at 6 months. An important facet of our study was the development and use of a novel QST modelling approach, allowing us to model all data together across modalities. We have not demonstrated significant differences in thermal thresholds between subjects with SCD and controls. Thermal thresholds were consistent over a 3- to 6-month period. Subjects on whom hydroxycarbamide (HC) was initiated shortly before or after baseline testing (new HC users) exhibited progressive decreases in thermal sensitivity from baseline to 6 months, suggesting that thermal testing may be sensitive to effective therapy to prevent vasoocclusive pain. These findings inform the use of QST as an endpoint in the evaluation of preventative pain therapies.

Establishing treatment fidelity in evidence-based parent training programs for externalizing disorders in children and adolescents.

The current review evaluates the use of treatment fidelity strategies in evidence-based parent training programs for treating externalizing disorders. We used a broad framework for evaluating treatment fidelity developed by the National Institutes of Health Treatment Fidelity Workgroup that includes the aspects of treatment design, treatment delivery, training providers, and assessment of participant receipt of treatment and enactment of treatment skills. Sixty-five articles reporting outcome trials of evidence-based parent training programs met inclusion criteria and were coded for treatment fidelity strategies. The mean adherence to fidelity strategies was .73, which was higher than two previous review studies employing this framework in the health literature. Strategies related to treatment design showed the highest mean adherence (.83), whereas training of providers and enactment of treatment skills had the lowest (.58). In light of an increasing emphasis on effectiveness and dissemination trials, the broader treatment fidelity framework as applied in this review focuses needed attention on areas often overlooked in fidelity practices, such as training providers and generalization of treatment skills. We discuss the strengths and limitations of fidelity practices in parent training studies, implications of these findings, and areas for future research.

Effects of a synbiotic on fecal quality, short-chain fatty acid concentrations, and the microbiome of healthy sled dogs.

BACKGROUND: Sled dogs commonly suffer from diarrhea. Although multiple etiologies exist there are limited field studies using synbiotics as a supplement to prevent or treat diarrhea. The objective of this study was to examine alterations in fecal quality, short-chain fatty acids (SCFA), and the fecal microbiome in two groups of training sled dogs fed a synbiotic or microcrystalline cellulose placebo. Twenty clinically healthy training sled dogs randomized into two cohorts (9 synbiotic-fed, 8 placebo-fed) for a 6 week prospective study were examined. Fecal pH and fecal short chain fatty acid (SCFA) concentrations were measured and tag-encoded FLX 16S rDNA amplicon pyrosequencing (bTEFAP) and quantitative real-time PCR were performed at baseline (10 d prior to the study) and after 2 weeks of treatment with a total treatment time of 6 weeks. Fecal scores for all dogs were assessed at baseline and every day for 6 wk after initiation of treatment. RESULTS: Alterations in the fecal microbiome were observed with a significant rise in Lactobacillaceae in the synbiotic group (P = 0.004) after 2 wk of treatment. A positive correlation was found between Lactobacillaceae and overall butyrate concentration (R = 0.62, p = 0.011) in all dogs. After 5 wk of treatment, there was an improved fecal score and fewer days of diarrhea (Χ2 = 5.482, P = 0.019) in the dogs given synbiotic, which coincided with a presumed contagious outbreak shared by all dogs in the study. CONCLUSIONS: Use of this synbiotic results in an increase in presumed beneficial bacterial flora of the host colon which was associated with a decrease in the prevalence of diarrhea in training sled dogs.

BCL-2 inhibitors sensitize therapy-resistant chronic lymphocytic leukemia cells to VSV oncolysis.

Many primary cancers including chronic lymphocytic leukemia (CLL) are resistant to vesicular stomatitis virus (VSV)-induced oncolysis due to overexpression of the antiapoptotic and antiautophagic members of the B-cell lymphoma-2 (BCL-2) family. In the present study, we investigated the mechanisms of CLL cell death induced as a consequence of VSV infection in the presence of BCL-2 inhibitors, obatoclax, and ABT-737 in primary ex vivo CLL patient samples. Microarray analysis of primary CD19⁺ CD5⁺ CLL cells treated with obatoclax and VSV revealed changes in expression of genes regulating apoptosis, the mechanistic target of rapamycin (mTOR) pathway, and cellular metabolism. A combined therapeutic effect was observed for VSV and BCL-2 inhibitors in cells from untreated patients and from patients unresponsive to standard of care therapy. In addition, combination treatment induced several markers of autophagy--LC3-II accumulation, p62 degradation, and staining of autophagic vacuoles. Inhibition of early stage autophagy using 3-methyladenine (3-MA) led to increased apoptosis in CLL samples. Mechanistically, a combination of BCL-2 inhibitors and VSV disrupted inhibitory interactions of Beclin-1 with BCL-2 and myeloid cell leukemia-1 (MCL-1), thus biasing cells toward autophagy. We propose a mechanism in which changes in cellular metabolism, coupled with pharmacologic disruption of the BCL-2-Beclin-1 interactions, facilitate induction of apoptosis and autophagy to mediate the cytolytic effect of VSV.

Targeted quantitative analysis of superoxide dismutase 1 in cisplatin-sensitive and cisplatin-resistant human ovarian cancer cells.

Protein quantification in a complex protein mixture presents a daunting task in biochemical analysis. Antibody-based immunoassays are traditional methods for protein quantification. However, there are issues associated with accuracy and specificity in these assays, especially when the changes are small (e.g., <2-fold). With recent developments in mass spectrometry, monitoring a selected peptide, thus protein, in a complex biological sample has become possible. In this study, we demonstrate a simple mass spectrometry-based method for selective measurement of a moderately low abundant protein, superoxide dismutase 1 (SOD1), in cisplatin-sensitive and cisplatin-resistant human ovarian cancer cells. Selected-reaction-monitoring (SRM) technology was employed to specifically analyze the target peptides in a pair of human ovarian cancer cell lines: 2008/2008-C13*5.25 (cisplatin-sensitive/cisplatin-resistant, respectively). The observed 1.47-fold higher expression in the resistant cell line is consistent with findings by other approaches. This robust liquid chromatography/mass spectrometry (LC/MS) method provides a powerful tool for targeted proteomic verification and/or validation studies.

Targeting superoxide dismutase 1 to overcome cisplatin resistance in human ovarian cancer.

PURPOSE: Clinical drug resistance to platinum-based chemotherapy is considered a major impediment in the treatment of human ovarian cancer. Multiple pathways associated with drug resistance have been suggested by many previous studies. Over expression of several key proteins involved in DNA repair, drug transport, redox regulation, and apoptosis has been recently reported by our group using a global quantitative proteomic profiling approach. Superoxide dismutase 1 (SOD1) is one of these proteins consistently over-expressed in cisplatin-resistant ovarian cancer cells as compared to their sensitive counterparts, but its precise role in drug resistance is yet to be defined. METHOD: In the current study, we examined the role of SOD1 in drug resistance by inhibiting its redox activity in cisplatin-resistant ovarian cancer cells using a small-molecule inhibitor, triethylenetetramine (TETA). The effect of TETA was determined by the cell proliferation assay, clonogenic cell survival assay, and SOD1 activity assay. RESULTS: The inhibition of the SOD1 activity enhanced the cisplatin sensitivity in the resistant cells supporting the hypothesis that SOD1 is a key determinant of cisplatin resistance and is an exploitable target to overcome cisplatin drug resistance. CONCLUSION: SOD1 plays an important role in cisplatin resistance and modulation of its activity may overcome this resistance and ultimately lead to improved clinical outcomes.

Label-free mass spectrometry-based protein quantification technologies in proteomic analysis.

Major technological advances have made proteomics an extremely active field for biomarker discovery and validation in recent years. These improvements have lead to an increased emphasis on larger scale, faster and more efficient methods for protein biomarker discoveries in human tissues, cells and biofluids. However, most current proteomic methodologies for biomarker discovery and validation are not highly automated and generally labour intensive and expensive. Improved automation as well as software programs capable of handling a large amount of data are essential in order to reduce the cost of discovery and increase the throughput. In this review, we will discuss and describe the label-free mass spectrometry-based protein quantification technologies and a case study utilizing one of these methods for biomarker discovery.

A comparative proteomic study to characterize the vinblastine resistance in human ovarian cancer cells.

Drug resistance is a major impediment to the successful treatment of human cancers, including ovarian cancer. Vinblastine (VLB), an antimicrotubule agent, is one of the chemotherapeutic drugs that exhibit resistance in ovarian cancer patients. To determine the protein factors that are involved in vinblastine resistance in human ovarian cancer cells, a combination of sample pre-fractionation and high-resolution 2-DE proteomic analysis was performed. Approximately 1200 proteins were detected and quantitatively compared in both nuclear/membrane and cytosolic fractions. Sixty-nine proteins from the nuclear/membrane fraction showed altered expression levels, whereas 59 were altered in the cytosolic fraction between SKOV3 (vinblastine-sensitive) and SKVLB (vinblastine-resistant) cell lines. These proteins include membrane-associated, chromatin remodeling, cytoskeletal, and microtubule-associated proteins as well as others that regulate signal transduction. This study not only demonstrates a novel understanding of the mechanism of drug resistance but also provides a valuable resource for future studies on drug resistance to vinblastine. In addition, it also represents a good example of how to increase the protein dynamic range and reduce sample complexity using currently available tools.

Intracellular signal transduction for migration and actin remodeling in vascular smooth muscle cells after sphingosylphosphorylcholine stimulation.

Molecular mechanisms underlying migration of vascular smooth muscle cells (VSMCs) toward sphingosylphosphorylcholine (SPC) were analyzed in light of the hypothesis that remodeling of the actin cytoskeleton should be involved. After SPC stimulation, mitogen-activated protein kinases (MAPKs), including p38 MAPK (p38) and p42/44 MAPK (p42/44), were found to be phosphorylated. Migration of cells toward SPC was reduced in the presence of SB-203580, an inhibitor of p38, but not PD-98059, an inhibitor of p42/44. Pertussis toxin (PTX), a Gi protein inhibitor, induced an inhibitory effect on p38 phosphorylation and VSMC migration. Myosin light chain (MLC) phosphorylation occurred after SPC stimulation with or without pretreatment with SB-203580 or PTX. The MLC kinase inhibitor ML-7 and the Rho kinase inhibitor Y-27632 inhibited MLC phosphorylation but only partially inhibited SPC-directed migration. Complete inhibition was achieved with the addition of SB-203580. After SPC stimulation, the actin cytoskeleton formed thick bundles of actin filaments around the periphery of cells, and the cells were surrounded by elongated filopodia, i.e., magunapodia. The peripheral actin bundles consisted of alpha- and beta-actin, but magunapodia consisted exclusively of beta-actin. Such a remodeling of actin was reversed by addition of SB-203580 and PTX, but not ML-7 or Y-27632. Taken together, our biochemical and morphological data confirmed the regulation of actin remodeling and suggest that VSMCs migrate toward SPC, not only by an MLC phosphorylation-dependent pathway, but also by an MLC phosphorylation-independent pathway.

A systems biology case study of ovarian cancer drug resistance.

In ovarian cancer treatment, the chemotherapy drug cisplatin often induce drug resistance after prolonged use, causing cancer relapse and the eventual deaths of patients. Cisplatin-induced drug resistance is known to involve a complex set of cellular changes but its molecular mechanism(s) remain unclear. In this study, we designed a systems biology approach to examine global protein level and network level changes by comparing Proteomics profiles between cisplatin-resistant cell lines and cisplatin-sensitive cell lines. First, we used an experimental proteomics method based on a Label-free Liquid Chromatography / Mass Spectrometry (LC/MS) platform to obtain a list of 119 proteins that are differentially expressed in the samples. Second, we expanded these proteins into a cisplatin-resistant activated sub-network, which consists of 1230 proteins in 1111 protein interactions. An examination of network topology features reveals the activated responses in the network are closely coupled. Third, we examined sub-network proteins using Gene Ontology categories. We found significant enrichment of proton-transporting ATPase and ATP synthase complexes in addition to protein binding proteins. Fourth, we examined sub-network protein interaction function categories using 2-dimensional visualization matrixes. We found that significant cellular physiological responses arise from endogenous, abiotic, and stress-related signals, which correlates well with known facts that internalized cisplatin cause DNA damage and induce cell stress. Fifth and finally, we developed a new visual representation structure for display of activated sub-networks using functional categories as network nodes and their crosstalk as network edges. This type of sub-network further shows that while cell communication and cell growth are generally important to tumor mechanisms, molecular regulation of cell differentiation and development caused by responses to genomic-wide stress seem to be more relevant to the acquisition of drug resistance.