Intermolecular interactions underlie protein/peptide phase separation irrespective of sequence and structure at crowded milieu.

Liquid-liquid phase separation (LLPS) has emerged as a crucial biological phenomenon underlying the sequestration of macromolecules (such as proteins and nucleic acids) into membraneless organelles in cells. Unstructured and intrinsically disordered domains are known to facilitate multivalent interactions driving protein LLPS. We hypothesized that LLPS could be an intrinsic property of proteins/polypeptides but with distinct phase regimes irrespective of their sequence and structure. To examine this, we studied many (a total of 23) proteins/polypeptides with different structures and sequences for LLPS study in the presence and absence of molecular crowder, polyethylene glycol (PEG-8000). We showed that all proteins and even highly charged polypeptides (under study) can undergo liquid condensate formation, however with different phase regimes and intermolecular interactions. We further demonstrated that electrostatic, hydrophobic, and H-bonding or a combination of such intermolecular interactions plays a crucial role in individual protein/peptide LLPS.

An Update on Potential Molecular Biomarkers of Dietary Phytochemicals Targeting Lung Cancer Interception and Prevention.

Since ancient times, dietary phytochemicals are known for their medicinal properties. They are broadly classified into polyphenols, terpenoids, alkaloids, phytosterols, and organosulfur compounds. Currently, there is considerable interest in their potential health effects against various diseases, including lung cancer. Lung cancer is the leading cause of cancer deaths with an average of five-year survival rate of lung cancer patients limited to just 14%. Identifying potential early molecular biomarkers of pre-malignant lung cancer cells may provide a strong basis to develop early cancer detection and interception methods. In this review, we will discuss molecular changes, including genetic alterations, inflammation, signal transduction pathways, redox imbalance, epigenetic and proteomic signatures associated with initiation and progression of lung carcinoma. We will also highlight molecular targets of phytochemicals during lung cancer development. These targets mainly consist of cellular signaling pathways, epigenetic regulators and metabolic reprogramming. With growing interest in natural products research, translation of these compounds into new cancer prevention approaches to medical care will be urgently needed. In this context, we will also discuss the overall pharmacokinetic challenges of phytochemicals in translating to humans. Lastly, we will discuss clinical trials of phytochemicals in lung cancer patients.

A common human MLKL polymorphism confers resistance to negative regulation by phosphorylation.

Across the globe, 2-3% of humans carry the p.Ser132Pro single nucleotide polymorphism in MLKL, the terminal effector protein of the inflammatory form of programmed cell death, necroptosis. Here we show that this substitution confers a gain in necroptotic function in human cells, with more rapid accumulation of activated MLKLS132P in biological membranes and MLKLS132P overriding pharmacological and endogenous inhibition of MLKL. In mouse cells, the equivalent Mlkl S131P mutation confers a gene dosage dependent reduction in sensitivity to TNF-induced necroptosis in both hematopoietic and non-hematopoietic cells, but enhanced sensitivity to IFN-ß induced death in non-hematopoietic cells. In vivo, MlklS131P homozygosity reduces the capacity to clear Salmonella from major organs and retards recovery of hematopoietic stem cells. Thus, by dysregulating necroptosis, the S131P substitution impairs the return to homeostasis after systemic challenge. Present day carriers of the MLKL S132P polymorphism may be the key to understanding how MLKL and necroptosis modulate the progression of complex polygenic human disease.

Evaluation of a retinol containing topical treatment to improve signs of neck aging.

BACKGROUND: The neck region is an area that can be indicative of signs of skin aging. A novel topical product that combines multiple active ingredients including retinol, tripeptide and glaucine was formulated to specifically target neck aging correction and complement post-procedure as part of an integrated skincare regimen. OBJECTIVES: To evaluate the efficacy of a topical neck treatment through clinical subject evaluation, in addition to ultrasound and biopsy assessment. METHODS: Evaluation for the efficacy of this novel topical product on improving the aging signs of neck skin was performed in multiple clinical trials. The first trial focused on clinical efficacy and included clinical assessment, subject questionnaires, ultrasound imaging and digital photographs. The second trial focused on biomarker analysis through skin biopsy. RESULTS: Data from the clinical trials showed that aging signs on the neck were significantly improved after 12 or 16 weeks of product usage. Changes were readily observed by clinical evaluators and participants. They were documented with digital photos, ultrasound images, and biomarker expression in the skin which clearly display the improvements. CONCLUSIONS: This novel topical product is effective in treating the aging signs on the neck skin and has been shown to provide statistically significant improvement on a myriad of neck aging attributes including fine lines/wrinkles, crepiness, laxity, and texture.

Amyloid fibril-based thixotropic hydrogels for modeling of tumor spheroids in vitro.

Biomaterials mimicking extracellular matrices (ECM) for three-dimensional (3D) cultures have gained immense interest in tumor modeling and in vitro organ development. Here, we introduce a new class of amyloid fibril-based peptide hydrogels as a versatile biomimetic ECM scaffold for 3D cell culture and homogenous tumor spheroid modeling. We show that these amyloid fibril-based hydrogels are thixotropic and allow cancer cell adhesion, proliferation, and migration. All seven designed hydrogels support 3D cell culture with five different cancer cell lines forming spheroid with necrotic core and upregulation of the cancer biomarkers. We further developed the homogenous, single spheroid using the drop cast method and the data suggest that all hydrogels support the tumor spheroid formation but with different necrotic core diameters. The detailed gene expression analysis of MCF7 spheroid by microarray suggested the involvement of pro-oncogenes and significant regulatory pathways responsible for tumor spheroid formation. Further, using breast tumor tissue from a mouse xenograft model, we show that selected amyloid hydrogels support the formation of tumor spheroids with a well-defined necrotic core, cancer-associated gene expression, higher drug resistance, and tumor heterogeneity reminiscent of the original tumor. Altogether, we have developed an easy-to-use, rapid, cost-effective, and scalable platform for generating in vitro cancer models for the screening of anti-cancer therapeutics and developing personalized medicine.

Charge and hydrophobicity of amyloidogenic protein/peptide templates regulate the growth and morphology of gold nanoparticles.

Biomolecules are known to interact with metals and produce nanostructured hybrid materials with diverse morphologies and functions. In spite of the great advancement in the principles of biomimetics for designing complex nano-bio structures, the interplay between the physical properties of biomolecules such as sequence, charge, and hydrophobicity with predictable morphology of the resulting nanomaterials is largely unknown. Here, using various amyloidogenic proteins/peptides and their corresponding fibrils in combination with different pH, we show defined principle for gold nanocrystal growth into triangular and supra-spheres with high prediction. Using a combination of different biophysical and structural techniques, we establish the mechanism of nucleation and crystal growth of gold nanostructures and show the effective isolation of intact nanostructures from amyloid templates using protein digestion. This study will significantly advance our design principle for bioinspired materials for specific functions with great predictability.

The use of, and outcomes for, inflammatory bowel disease services during the Covid-19 pandemic: a nationwide observational study.

BACKGROUND: Inflammatory bowel disease (IBD) services have been particularly affected by the Covid-19 pandemic. Delays in referral to secondary care and access to investigations and surgery have been exacerbated. AIMS: To investigate the use of and outcomes for emergency IBD care during the Covid-19 pandemic. METHODS: Nationwide observational study using administrative data for England (2015-2020) comparing cohorts admitted from 1 January 2015, to 31 January 2020 (pre-pandemic) and from 1 February 2020, to 31 January 2021 (pandemic). Autoregressive integrated moving average forecast models were run to estimate the counterfactual IBD admissions and procedures for February 2020 to January 2021. RESULTS: Large decreases in attendances to hospital for emergency treatment were observed for both acute ulcerative colitis (UC, 16.4%) and acute Crohn's disease (CD, 8.7%). The prevalence of concomitant Covid-19 during the same episode was low [391/16 494 (2.4%) and 349/15 613 (2.2%), respectively]. No significant difference in 30-day mortality was observed. A shorter median length of stay by 1 day for acute IBD admissions was observed (P < 0.0001). A higher rate of emergency readmission within 28 days for acute UC was observed (14.1% vs 13.4%, P = 0.012). All IBD procedures and investigations showed decreases in volume from February 2020 to January 2021 compared with counterfactual estimates. The largest absolute deficit was in endoscopy (17 544 fewer procedures, 35.2% reduction). CONCLUSION: There is likely a significant burden of untreated IBD in the community. Patients with IBD may experience clinical harm or protracted decreases in quality of life if care is not prioritised.

A community-based partnership collaborative practice agreement project to disseminate and implement evidence-based practices in community pharmacy.

BACKGROUND AND PURPOSE: The objective of this study is to explore the impact of an educational intervention including (1) collaborative practice agreement (CPA)-focused lecture and (2) a student project in partnership with a community-based pharmacy on dissemination and implementation of CPAs in community pharmacy practice. EDUCATIONAL ACTIVITY AND SETTING: A CPA-focused classroom lecture and related project were given to five sequential years of second-year pharmacy students enrolled in a community pharmacy elective course. Community pharmacist feedback provided at the completion of the project was compared to present-day data collected via telephone survey. Responses for each survey question were summarized using frequencies, and chi-square analysis was conducted to assess the association between pharmacist perceptions at the time of the project vs. the present day. FINDINGS: Forty-seven projects representing 47 unique CPAs were completed over five sequential years of the course. The most commonly targeted disease states by CPAs were influenza/streptococcus pharyngitis (25.5%), therapeutic interchange (6.4%), oral contraceptives (6.4%), and tobacco cessation (6.4%). Pharmacists noted that students facilitated their progress toward CPA implementation by serving as a means for CPA law education (23.4%) and by saving time in CPA development (12.8%). SUMMARY: This study demonstrated that a student project partnering with a local community pharmacy improved knowledge dissemination but did not have any direct impact on implementation of CPAs in a community pharmacy. However, students may have a role in dissemination and implementation of evidence-based practices, such as CPAs, when appropriately matched to setting-specific implementation barriers.

Direct evidence of cellular transformation by prion-like p53 amyloid infection.

Tumor suppressor p53 mutations are associated with more than 50% of cancers. Aggregation and amyloid formation of p53 is also implicated in cancer pathogenesis, but direct evidence for aggregated p53 amyloids acting as an oncogene is lacking. Here, we conclusively demonstrate that wild-type p53 amyloid formation imparts oncogenic properties to non-cancerous cells. p53 amyloid aggregates were transferred through cell generations, contributing to enhanced survival, apoptotic resistance with increased proliferation and migration. The tumorigenic potential of p53 amyloid-transformed cells was further confirmed in mouse xenografts, wherein the tumors showed p53 amyloids. p53 disaggregation rescued the cellular transformation and inhibited tumor development in mice. We propose that wild-type p53 amyloid formation contributes to tumorigenesis and can be a potential target for therapeutic intervention. This article has an associated First Person interview with the first author of the paper.

Severe Acute Respiratory Syndrome Coronavirus 2 Induced Focal Segmental Glomerulosclerosis.

Focal segmental glomerulosclerosis (FSGS) is a common cause of nephrotic syndrome affecting adults and children. Collapsing focal segmental glomerulosclerosis (FSGS), one of five histologic variants of FSGS is described as segmental or global collapse and sclerosis of the glomerular tufts and has been frequently associated with human immunodeficiency virus-associated nephropathy (HIVAN). Its association with other viral and non-viral causes, medications and other disease states has since been established. Due to its resistance to therapy, rapid progression to end-stage renal disease (ESRD) and overall poorer prognosis, identification with electron microscopy examination of the kidney biopsy sample is required during evaluation.

Mutational Landscape for Indian Hereditary Breast and Ovarian Cancer Cohort Suggests Need for Identifying Population Specific Genes and Biomarkers for Screening.

BACKGROUND: Breast and ovarian cancers are the most prevalent cancers and one of the leading causes of death in Indian women. The healthcare burden of breast and ovarian cancers and the rise in mortality rate are worrying and stress the need for early detection and treatment. METHODS: We performed amplicon sequencing of 144 cases who had breast/ovarian cancer disease (total 137 cases are patients and seven are tested for BRCA1/2 carrier) Using our custom designed gene panel consisting of 14 genes, that are associated with high to moderate risk of breast and ovarian cancers. Variants were called using Torrent Variant Caller and were annotated using ThermoFisher's Ion Reporter software. Classification of variants and their clinical significance were identified by searching the variants against ClinVar database. RESULTS: From a total of 144 cases, we were able to detect 42 pathogenic mutations in [40/144] cases. Majority of pathogenic mutations (30/41) were detected in BRCA1 gene, while (7/41) pathogenic mutations were detected in BRCA2 gene, whereas, (2/41) pathogenic mutations were detected in TP53 gene and BRIP1, PALB2, and ATM genes respectively. So, BRCA genes contributed 88.09% of pathogenic mutations, whereas non-BRCA genes contributed 11.91% of pathogenic mutations. We were also able to detect 25 VUS which were predicted to be damaging by in silico prediction tools. CONCLUSION: Early detection of cancers in the Indian population can be done by genetic screening using customized multi-gene panels. Indications of our findings show that in the Indian population, apart from the common BRCA genes, there are other genes that are also responsible for the disease. High frequency mutations detected in the study and variants of uncertain significance predicted to be damaging by in silico pathogenicity prediction tools can be potential biomarkers of hereditary breast and ovarian cancer in Indian HBOC patients.

A Unique Case of Alpha-Fetoprotein-Negative Hepatoid Adenocarcinoma of the Stomach With Associated Signet Ring Cell Histological Features and Linitis Plastica.

Hepatoid adenocarcinoma of the stomach (HAS), a rare and unique histological subtype of gastric cancer, accounts for less than 1.5% of all gastric cancers. Historically, this subtype is found to have a poor prognosis in comparison to other types of gastric cancer. While the diagnosis is made based on pathological findings, most cases described in the literature are associated with elevated alpha-fetoprotein (AFP) levels. We present a case of AFP-negative HAS with additional unique pathologic findings of signet ring cells which has been reported only once in the literature. Given the rare and late presentation of the disease, AFP-negative HAS should be included in the differential diagnosis in patients with suspicion for gastric cancer.

The Importance of Risk-Reducing Prophylactic Mastectomy in Breast Cancer (BRCA) Carriers: A Case Report.

Genetic changes along with environmental exposures can play a role in the development of cancer. Individuals with significant risk factors for breast cancer including family history should be encouraged to undergo genetic testing along with breast cancer screening at an early age. Individuals who test positive for the breast cancer (BRCA) 1 or 2 gene can discuss potential risk-reducing management options, such as risk-reducing prophylactic mastectomy, with their physicians in order to protect them from the long-term consequences of developing breast cancer.

Decreasing Inappropriate Use of Rasburicase to Promote Cost-Effective Care.

BACKGROUND: Rasburicase is a recommended treatment of tumor lysis syndrome and patients at high-risk for developing tumor lysis syndrome. Unfortunately, it is expensive, and unnecessary use raises costs of care. METHODS: Plan, Do, Study, Act methodology was used to decrease the inappropriate use of rasburicase. In the Plan phase, a multidisciplinary quality improvement team reviewed the rasburicase ordering process and its prescription patterns at Parkland Health and Hospital System between October 2015 and September 2017 to determine appropriate interventions for improvement. In the Do phase, interventions were deployed to improve rasburicase prescriptions. In the Study phase, the team reviewed the rasburicase orders and appropriateness from February 2018 to October 2018. During the Act phase, the interventions were found to be successful, and the process changes were solidified. RESULTS: At baseline, 65 doses of rasburicase were administered during the 2-year baseline period, 21 of these (32.3%) were inappropriate. Review of the ordering process identified pitfalls: one-click ready-to-sign order, fixed default dose, no hard-stop alert requiring physicians to review and confirm appropriate indications, and lack of secondary pharmacy review. We aimed to reduce the percentage of inappropriate rasburicase orders from a baseline of 32.3% to 10% over 3 months. In February 2018, we implemented the interventions, which resulted in reduction in inappropriate rasburicase use, with only a single inappropriate order placed in 7 months postintervention. CONCLUSION: A multidisciplinary approach and classic quality improvement methodology enabled us to reduce inappropriate rasburicase use. Straightforward electronic medical record interventions and secondary pharmacy review are effective in addressing overuse.

Amyloid Fibrils: Versatile Biomaterials for Cell Adhesion and Tissue Engineering Applications.

Extracellular matrices (ECM) play an enormous role in any living system, controlling various factors and eventually fates of cells. ECM regulates cell fate by providing constant exogenous signals altering intracellular signal transduction for diverse pathways including proliferation, migration, differentiation, and apoptosis. Biomaterial scaffolds are designed to mimic the natural extracellular matrix such that the cells could recapitulate natural events alike their natural niche. Therefore, the success of tissue engineering is largely dependent on how one can engineer the natural matrix properties at nanoscale precision. In this aspect, several recent studies have suggested that, as long as amyloid fibrils are not toxic, they can be utilized for cell adhesion and tissue engineering applications due to its ECM mimetic surface topography and ability to mediate active cell adhesion via focal adhesions. Although historically associated with human diseases, amyloids have presently emerged as one of the excellent biomaterials evolved in nature. In this review, we focus on the recent advances of amyloid-based biomaterials for cell adhesion and tissue engineering applications.

Enhancing Nurses' Oral Therapy Practice in 4 Latin American Countries: A Collaborative and Participatory Approach.

BACKGROUND: Oral therapy (OT) use for cancer is increasing globally. Yet, nurses in 4 Latin American countries lacked knowledge and educational opportunities to safely care for people receiving OTs. Global partnerships to contextualize education and create local capacity may enhance nursing practice. OBJECTIVE: Within 4 Latin American countries, this study aims to (1) develop, deliver, and evaluate an OT cancer nursing education program and (2) evaluate the feasibility and efficacy of using an integrated knowledge translation (iKT) framework to develop the program and foster nurses' capacity for OT care. METHODS: Using the iKT framework, a "train the trainer" model was used to develop, contextualize, pilot test, implement, and evaluate the OT education program. An online survey evaluated nurses' perceived benefits, ease of use, barriers, facilitators, and recommendations for improvement. Nurses' self-reported OT practices were evaluated 9 months after the final workshop. RESULTS: One hundred nineteen nurses across 4 countries participated in a pilot and/or final OT educational workshop, facilitated by 6 local nurse champions. The nurse champions found the program easy to use and modify. Participants reported using the curriculum to teach other nurses and patients and networking opportunities for problem solving. Barriers included nurses' role clarity and time for education. CONCLUSIONS: The iKT approach was an effective method to develop the OT curriculum and build OT capacity among nurses and leaders within the 4 countries. IMPLICATIONS FOR PRACTICE: The iKT approach may be useful in low- or middle-income countries to enhance nursing education and practice. Future OT education projects should strengthen strategies for ongoing support after education intervention.

The preemptive use of extracorporeal membrane oxygenation to facilitate high risk percutaneous cardiac interventions: left main percutaneous coronary interventions and percutaneous balloon aortic valvuloplasty.

As percutaneous cardiac interventions continue to evolve, high-risk procedures are being performed on patients deemed too ill for surgery. What were once considered lower-risk procedures compared with open cardiac interventions may no longer be so because of the complex nature of the interventions and the complex comorbidities of the patients on whom these are being performed. We present a case involving high-risk percutaneous cardiac interventions (left main coronary angioplasty and aortic balloon valvuloplasty), facilitated by the use of extracorporeal membrane oxygenation in a critically ill patient with severe aortic stenosis, left main coronary disease, and ischemic cardiomyopathy.

The effect of routine intraoperative transesophageal echocardiography on surgical management.

OBJECTIVE: To assess the effects of routine intraoperative transesophageal echocardiography (TEE) on surgical management of patients undergoing all types of cardiac surgery. DESIGN: Prospective, observational. SETTING: A single-institution, clinical investigation, university-affiliated hospital. PARTICIPANTS: Two hundred eighty-three consecutive patients undergoing cardiac surgery. INTERVENTIONS: A comprehensive TEE examination was performed in every patient after the induction of anesthesia. An appropriate surgical plan was then developed. A focused TEE examination was also performed at the conclusion of surgery. Whether or not TEE findings represented new information and whether or not this new information altered surgical management was documented. MEASUREMENTS AND MAIN RESULTS: There were 106 new TEE findings in 87 patients (31%). Half of the new findings involved the mitral valve, and a quarter involved the tricuspid valve. The new TEE information altered surgical management 77 ways in 71 patients (25%). Half of the altered surgical managements involved the mitral valve, and a third involved the tricuspid valve. In 8 patients (3%), TEE information influenced decisions regarding use/nonuse of cardiopulmonary bypass (CPB). In 2 patients, TEE examination after the separation from CPB prompted reinitiation of CPB. In 1 patient, TEE examination after the induction of general anesthesia prompted cancellation of surgery. CONCLUSIONS: The routine use of TEE during cardiac surgery revealed new cardiac pathology in 1 of every 3 patients and led to altered surgical management in 1 of every 4 patients. TEE information also influenced decisions regarding use/nonuse of CPB in 3% of patients. Thus, the authors suggest that intraoperative TEE should be used routinely in all patients undergoing cardiac surgery.

Transoesophageal echocardiography and central line insertion.

We investigated the potential utility of transoesophageal echocardiography (TOE) in facilitating central venous catheter (CVC) insertion in patients undergoing cardiac surgery. Thirty five patients undergoing elective cardiac surgery and CVC insertion were prospectively included in the observational, single-centre clinical investigation. Following induction of general anaesthesia and tracheal intubation, the TOE probe was inserted and the bicaval view obtained prior to CVC insertion (site at discretion of the anaesthesiologist). Prospectively collected data included site and sequence of CVC insertion attempts, information regarding ease of guidewire insertion, whether or not guidewire was visualized via TOE, and other pertinent information. In 1 patient, the TOE bicaval view could not be readily obtained because of right atrial (RA) distortion. In 31 patients, the TOE bicaval view was obtained and CVC access was successful at the site of first choice (guidewire visualized in all). Three patients had noteworthy CVC insertions. In one, CVC insertion was difficult despite visualization ofguidewire in the RA. In another, multiple guidewire insertions met with substantial resistance and without visualization of guidewire in the RA. One patient was found to have an unanticipated large mobile superior vena cava thrombus that extended into the RA, which changed clinical management by prompting initial CVC insertion into the femoral vein (potentially avoiding morbidity associated with thrombus dislodgement). Our prospective observational clinical study indicates that routine use of TOE during CVC insertion may help avoid potential complications associated with this intervention. If both CVC insertion and TOE are going to be used in the same patient, the benefits of TOE should be maximized by routine visualization of the bicaval view during guidewire insertion.