Phenotypic heterogeneity drives differential disease outcome in a mouse model of triple negative breast cancer.

The triple negative breast cancer (TNBC) subtype is one of the most aggressive forms of breast cancer that has poor clinical outcome and is an unmet clinical challenge. Accumulating evidence suggests that intratumoral heterogeneity or the presence of phenotypically distinct cell populations within a tumor play a crucial role in chemoresistance, tumor progression and metastasis. An increased understanding of the molecular regulators of intratumoral heterogeneity is crucial to the development of effective therapeutic strategies in TNBC. To this end, we used an unbiased approach to identify a molecular mediator of intratumoral heterogeneity in breast cancer by isolating two tumor cell populations (T1 and T2) from the 4T1 TNBC model. Phenotypic characterization revealed that the cells are different in terms of their morphology, proliferation and self-renewal ability in vitro as well as primary tumor formation and metastatic potential in vivo. Bioinformatic analysis followed by Kaplan Meier survival analysis in TNBC patients identified Metastasis associated colon cancer 1 (Macc1) as one of the top candidate genes mediating the aggressive phenotype in the T1 tumor cells. The role of Macc1 in regulating the proliferative phenotype was validated and taken forward in a therapeutic context with Lovastatin, a small molecule transcriptional inhibitor of Macc1 to target the T1 cell population. This study increases our understanding of the molecular underpinnings of intratumoral heterogeneity in breast cancer that is critical to improve the treatment of women currently living with the highly aggressive TNBC subtype.

Role of yoga in stress management and implications in major depression disorder.

Major Depressive Disorder (MDD) is one of the leading causes of disability affecting more than 340 million people and second largest contributor to global burden of disease. Chronic stress is a common risk factor and important contributor for MDD. Stress could be defined as the "perceived inability to cope". Stressful life events are shown to provoke a sequence of psychological and physiological adjustments including nervous, endocrine and immune mechanisms. Stress can lead to elevation of a variety of inflammatory cytokines and stress hormones, can cause autonomic dysfunction and imbalance in neurotransmitters. Yoga can reduce depressive symptoms by alleviating stress. Studies have shown that yoga can reduce inflammation, maintain autonomic balance and also has a role in maintaining the neurotransmitters. It has role on hypothalamic-pituitary-adrenal (HPA) axis, the peripheral nervous system including GABA, limbic system activity, inflammatory and endocrine responses. Yoga along with antidepressants can help in reducing the depressive symptoms in patient with MDD. Yoga is an ideal complementary and alternative therapy for mental health disorders.

Cost-effective and reliable genomic DNA extraction from plant seedlings for high-throughput genotyping in seed industries.

Genetic purity of seeds is one of the critical aspects in the seed industry. Molecular seed testing laboratories are utilizing PCR based diagnostic tools for genetic purity analysis. High quality DNA is an essential prerequisite for such analyses. Here, we demonstrate a robust and inexpensive DNA extraction method to isolate genomic DNA from variety of crops. Current method (M2) was compared with four commonly used DNA isolation methods for PCR-based genetic characterization and High Resolution Melt (HRM) based hybridity analysis of cotton, okra, tomato and maize using SSR markers. DNA extracted through current method showed excellent yield and quality as compared to other methods. High quality, PCR ready DNA was isolated within 30-50 min and displayed best results for genetic purity analysis using HRM. In contrast, several genomic DNA samples extracted using other methods were found unsuitable for HRM analysis. Our method can be a perfect choice in seed industry, where thousands of samples are processed every day. Notably, using our method single technician can extract DNA from 96 leaf samples within 30-50 min, at a cost of only $0.11/sample. Overall, current DNA extraction method is a reliable and cost-effective solution for large-scale genotyping experiments in the agricultural industry.

The Gut Microbiota: a Novel Player in the Pathogenesis of Uterine Fibroids.

Uterine fibroid is a common gynecological disorder that affects women of reproductive age and has emerged as a major public health concern. The symptoms have a negative influence on both their physical health and quality of life. The cost of treatment has a significant impact on the disease's burden. Even though its origin is uncertain, estrogen is thought to be a key player in fibroid pathophysiology. Many theories, including those based on genetic and environmental factors, explain what causes hyper-estrogenic condition in fibroid patients. One such possibility that is currently being explored is the hypothesis that an altered gut microbiome can contribute to the development of diseases characterized by estrogen dominance. Gut dysbiosis is often a "hot area" in the health sciences. According to a recent study, uterine fibroid patients have altered gut microbiome. A variety of risk factors influence both fibroid development and gut homeostasis. Diet, lifestyle, physical activity, and environmental contaminants have an impact on estrogen and the gut flora. A better understanding of uterine fibroids' pathophysiology is required to develop effective preventative and treatment options. A few ways by which the gut microbiota contributes to UF include estrogen, impaired immune function, inflammation, and altered gut metabolites. Therefore, in the future, while treating fibroid patients, various strategies to deal with changes in the gut flora may be advantageous. For developing suggestions for clinical diagnosis and therapy, we reviewed the literature on the relationship between uterine fibroids and the gut microbiota.

Patient stratification using point of care biomarkers in dry eye disease.

With changes in lifestyle, such as the increasing use of digital screens and rising demand for refractive surgery, dry eye disease has become increasingly prevalent in recent times. While we are equipped with a number of diagnostic modalities and a myriad of treatment forms, ranging from topical medication to procedural therapies, the condition remains an enigma in terms of varied patient satisfaction. An understanding of the molecular basis of a disease may open up new avenues in the customization of its treatment. We attempt to simplify this in the form of a stepwise protocol to incorporate biomarker assays in dry eye management.

Development of Deep Belief Network for Tool Faults Recognition.

The controlled interaction of work material and cutting tool is responsible for the precise outcome of machining activity. Any deviation in cutting parameters such as speed, feed, and depth of cut causes a disturbance to the machining. This leads to the deterioration of a cutting edge and unfinished work material. Recognition and description of tool failure are essential and must be addressed using intelligent techniques. Deep learning is an efficient method that assists in dealing with a large amount of dynamic data. The manufacturing industry generates momentous information every day and has enormous scope for data analysis. Most intelligent systems have been applied toward the prediction of tool conditions; however, they must be explored for descriptive analytics for on-board pattern recognition. In an attempt to recognize the variation in milling operation leading to tool faults, the development of a Deep Belief Network (DBN) is presented. The network intends to classify in total six tool conditions (one healthy and five faulty) through image-based vibration signals acquired in real time. The model was designed, trained, tested, and validated through datasets collected considering diverse input parameters.

Pharmacological attenuation of melanoma by tryptanthrin pertains to the suppression of MITF-M through MEK/ERK signaling axis.

Melanoma is the most aggressive among all types of skin cancers. The current strategies against melanoma utilize BRAFV600E, as a focal point for targeted therapy. However, therapy resistance developed in melanoma patients against the conventional anti-melanoma drugs hinders the ultimate benefits of targeted therapies. A major mechanism by which melanoma cells attain therapy resistance is via the activation of microphthalmia-associated transcription factor-M (MITF-M), the key transcription factor and oncogene aiding the survival of melanoma cells. We demonstrate that tryptanthrin (Tpn), an indole quinazoline alkaloid, which we isolated and characterized from Wrightia tinctoria, exhibits remarkable anti-tumor activity towards human melanoma through the down-regulation of MITF-M. Microarray analysis of Tpn-treated melanoma cells followed by a STRING protein association network analysis revealed that differential expression of genes in melanoma converges at MITF-M. Furthermore, in vitro and in vivo studies conducted using melanoma cells with differential MITF-M expression status, endogenously or ectopically, demonstrated that the anti-melanoma activity of Tpn is decisively contingent on its efficacy in down-regulating MITF-M expression. Tpn potentiates the degradation of MITF-M via the modulation of MEK1/2-ERK1/2-MITF-M signaling cascades. Murine models demonstrate the efficacy of Tpn in attenuating the migration and metastasis of melanoma cells, while remaining pharmacologically safe. In addition, Tpn suppresses the expression of mutated BRAFV600E and inhibits Casein Kinase 2α, a pro-survival enzyme that regulates ERK1/2 homeostasis in many tumor types, including melanoma. Together, we point to a promising anti-melanoma drug in Tpn, by virtue of its attributes to impede melanoma invasion and metastasis by attenuating MITF-M.

A Core-Linker-Polyamine (CLP) Strategy Enables Rapid Discovery of Antileishmanial Aminoalkylquinolinecarboxamides That Target Oxidative Stress Mechanism.

A Core-Linker-Polyamine (CLP) strategy has been exploited to develop new antileishmanial agents. It involves the linker-based assembly of alkyl-polyamine side chain as a potential pharmacophore motif with a privileged heterocyclic motif, 4-arylquinoline. A series of aminoalkyl 4-arylquinoline-2-carboxamides and their analogs were synthesized and tested against L. donovani promastigotes. Among all synthesized derivatives, 10 compounds showed significant antipromastigote activities with more efficacy (IC50 : 4.75-8 µM) than an antileishmanial oral drug Miltefosine (IC50 : 8.9±1.55 µM). Most active aminoalkyl-quinoline-carboxamides 9 a and 9 b, displayed negligible cytotoxicity towards human monocytic (THP-1) macrophages. The compounds show antileishmanial activity by generating mitochondrial superoxide radicals. However, they did not show interference with trypanothione reductase, a redox enzyme of Leishmania. Significant change in the morphology of the L. donovani promastigote by the compounds was observed. The Structure-activity relationship analysis suggest the pharmacophoric importance of alkylpolyamine and carboxamide motifs. In silico evaluation indicated that the investigated active molecules 9 a and 9 b possess important drug-likeness, physicochemical and pharmacokinetic-relevant properties.

A Mixed-Methods Examination of the Impact of the Partnerships to Improve Community Health Produce Prescription Initiative in Northeastern North Carolina.

OBJECTIVE: To conduct a mixed-methods examination of the impact of the Partnerships to Improve Community Health produce prescription initiative in northeastern North Carolina. DESIGN: Quantitative surveys were conducted among participants before and after the distribution of produce prescription vouchers. Univariate statistics were used to describe the participant population, and paired t tests were used to examine change in fruit and vegetable intake. Qualitative, in-depth telephone interviews were conducted among participants, health educators, and food retailers and coded for themes. SETTING: Eight health promotion programs, 2 food pantries, and 11 food retailers. PARTICIPANTS: In each health promotion program or food pantry, between 6 and 97 participants were enrolled. INTERVENTION: Produce prescription vouchers were distributed to participants and redeemed at local food retailers. MAIN OUTCOME MEASURE S: An increase in local fruit and vegetable purchasing and consumption. RESULTS: Of the produce prescription participants who completed the baseline survey (n = 93), 86% were female, 64% were African American, and 68% were food insecure. The voucher redemption rate was 18%. The majority of participants indicated that they visit farmers' markets more now than before the produce prescription initiative, that shopping at the farmers' market made it easy to include more fresh produce in their family's diet, and that they tried a new farmers' market because of the produce prescription initiative. All health educators and food retailers who participated felt that the initiative benefited their program or operation and were willing to partner with the program again. CONCLUSIONS: While redemption rates were lower than anticipated, the produce prescription initiative had positive impacts on participants' local fruit and vegetable purchasing and consumption. Because of COVID-19, the initiative was not implemented until late in the North Carolina produce season. Moving forward, the program will start earlier and work with local food retailers to connect with their communities to increase redemption rates.

B vitamin intakes modify the association between particulate air pollutants and incidence of all-cause dementia: Findings from the Women's Health Initiative Memory Study.

INTRODUCTION: Particulate air pollutants may induce neurotoxicity by increasing homocysteine levels, which can be lowered by high B vitamin intakes. Therefore, we examined whether intakes of three B vitamins (folate, B12 , and B6 ) modified the association between PM2.5 exposure and incidence of all-cause dementia. METHODS: This study included 7183 women aged 65 to 80 years at baseline. B vitamin intakes from diet and supplements were estimated by food frequency questionnaires at baseline. The 3-year average PM2.5 exposure was estimated using a spatiotemporal model. RESULTS: During a mean follow-up of 9 years, 342 participants developed all-cause dementia. We found that residing in locations with PM2.5 exposure above the regulatory standard (12 µg/m3 ) was associated with a higher risk of dementia only among participants with lower intakes of these B vitamins. DISCUSSION: This is the first study suggesting that the putative neurotoxicity of PM2.5 exposure may be attenuated by high B vitamin intakes.

Polarity Based Model for Guiding Medical School Strategy During Crisis - A Cross Sectional Qualitative Study.

BACKGROUND: Crises in academia can best be dealt with as a polarity that needs to be leveraged rather than a problem that needs to be solved. This work aimed at utilizing the Polarity Approach for Continuity and Transformation (PACT)™ to establish a guide for medical schools during times of crisis to minimize the effect of crisis-driven decisions on strategic growth. SUBJECTS AND METHODS: A qualitative study following the 5-Steps of the PACT process was conducted. A virtual mapping session was held with 108 medical educators from 22 countries to determine the upsides and downsides of strategic orientation and crisis management subsequently. RESULTS: Four polarity maps were generated identifying four tension areas; University reputation, mission, teams, and individuals followed by a 72-item assessment and another mapping session to map the warning signs and action steps. A comparison between private school scores and the whole cohort of respondents showed that private schools had the least problems in team-oriented work. CONCLUSION: This study highlighted the importance of taking measures to communicate the mission and supporting team functions inside universities either by enhancing resources or utilizing time and effort-saving strategies.

The Role of Temperature in Modifying the Risk of Ozone-Attributable Mortality under Future Changes in Climate: A Proof-of-Concept Analysis.

Air pollution risk assessments typically estimate ozone-attributable mortality counts using concentration-response (C-R) parameters from epidemiologic studies that treat temperature as a potential confounder. However, some recent epidemiologic studies have indicated that temperature can modify the relationship between short-term ozone exposure and mortality, which has potentially important implications when considering the impacts of climate change on public health. This proof-of-concept analysis quantifies counts of temperature-modified ozone-attributable mortality using temperature-stratified C-R parameters from a multicity study in which the pooled ozone-mortality effect coefficients change in concert with daily temperature. Meteorology downscaled from two global climate models is used with a photochemical transport model to simulate ozone concentrations over the 21st century using two emission inventories: one holding air pollutant emissions constant at 2011 levels and another accounting for reduced emissions through the year 2040. The late century climate models project increased summer season temperatures, which in turn yields larger total counts of ozone-attributable deaths in analyses using temperature-stratified C-R parameters compared to the traditional temperature confounder approach. This analysis reveals substantial heterogeneity in the magnitude and distribution of the temperature-stratified ozone-attributable mortality results, which is a function of regional variability in both the C-R relationship and the model-predicted temperature and ozone.

Dietary and socioeconomic risk factors for fumonisin exposure among women of reproductive age in 18 municipalities in Guatemala from 2013 to 2014.

Fumonisin exposure is common in populations where maize is a dietary staple, such as in Guatemala, and has been associated with negative health outcomes including neural tube defects. The objective of this study was to estimate fumonisin B1 (FB1) exposure among Guatemalan reproductive-age women and develop a better understanding of the dietary and sociodemographic risk factors for exposure. A cross-sectional study in 18 municipalities in Guatemala was conducted. Midwives and study nurses enrolled consenting women and collected individual and household demographic and socioeconomic data. A food frequency questionnaire was administered to estimate quantity and types of food products consumed. A urine sample was collected and urinary fumonisin B1 (uFB1) concentration was measured. A univariable analysis was conducted to identify predictors of low/high uFB1. Multivariable logistic regression was used to calculate adjusted odds ratios (ORs) and 95% confidence intervals (CIs). In total, 775 women had analyzable urine samples. Higher uFB1 levels were associated with speaking Mayan (OR = 2.33, 95% CI:1.44-3.77), less than high school education (OR = 1.61, 95% CI:1.12-2.30), increasing dietary proportion of maize-based foods (OR = 1.02, 95% CI:1.01-1.03), and consumption of tostadas (fried tortillas) (OR = 1.11, 95% CI:1.02-1.22). Lower uFB1 levels were associated with consumption of highly processed maize-based foods (OR = 0.93, 95% CI:0.87-0.99). Tortillas were the most frequently consumed maize-based food among study participants and significantly associated with high uFB1 exposure in the univariable but not multivariable analysis. Consumption of >4,750 grams/week of maize-based foods, >5,184 g/week of locally produced maize-based foods, and >110 servings/week of tortillas were also significantly associated with high uFB1 exposure in univariable analysis. Populations with low socioeconomic status/education levels and high consumption of maize-based foods had higher fumonisin exposure. Interventions aimed at reducing the risk of exposure to mycotoxins through maize in Guatemala, including the increased consumption of non-maize-based foods, should be further explored.

Knowledge and Attitude Among Various Dental Specialities Regarding Nasoalveolar Molding Procedure in Cleft Patients-A Cross-Sectional Study.

BACKGROUND: Nasoalveolar molding (NAM) is a presurgical orthopedic procedure, mainly focusing upon the aesthetic outcome of the treatment goal by reducing the number of reconstructive surgeries performed later for the purpose of esthetics. It was reported a very small proportion of pediatric dentists are providing presurgical intervention and the practice of NAM still side stepping among various dental practitioners. OBJECTIVES: The study was aimed to assess the knowledge and attitude among various dental specialties regarding NAM procedure in cleft patients. MATERIALS AND METHODS: A questionnaire-based cross-sectional survey was carried out among 180 dental professionals (pedodontists, orthodontists, prosthodontists, oral surgeons). A total of 180 questionnaires were prepared, out of which 98 were filled through direct personal interview and 82 were circulated via Google forms. Results were analyzed using chi-square test, Mann-Whitney U test, and Kruskal-Wallis test. RESULTS: There was statistically significant difference in the cumulative knowledge scores between specialty with P < .001; 99% of the participants think one should opt for NAM before cleft lip and palate surgery and 39% participants prefer pedodontists to carry out NAM procedure. CONCLUSION: Dental specialists in India have a positive attitude toward presurgical orthopedics. Majority of them agreed that there is great improvement in the aesthetic outcomes of surgeries after NAM intervention.

Lineage Plasticity in Cancer: The Tale of a Skin-Walker.

Lineage plasticity, the switching of cells from one lineage to another, has been recognized as a cardinal property essential for embryonic development, tissue repair and homeostasis. However, such a highly regulated process goes awry when cancer cells exploit this inherent ability to their advantage, resulting in tumorigenesis, relapse, metastasis and therapy resistance. In this review, we summarize our current understanding on the role of lineage plasticity in tumor progression and therapeutic resistance in multiple cancers. Lineage plasticity can be triggered by treatment itself and is reported across various solid as well as liquid tumors. Here, we focus on the importance of lineage switching in tumor progression and therapeutic resistance of solid tumors such as the prostate, lung, hepatocellular and colorectal carcinoma and the myeloid and lymphoid lineage switch observed in leukemias. Besides this, we also discuss the role of epithelial-mesenchymal transition (EMT) in facilitating the lineage switch in biphasic cancers such as aggressive carcinosarcomas. We also discuss the mechanisms involved, current therapeutic approaches and challenges that lie ahead in taming the scourge of lineage plasticity in cancer.

A four-year observational study to examine the dietary impact of the North Carolina Healthy Food Small Retailer Program, 2017-2020.

BACKGROUND: The North Carolina (NC) Healthy Food Small Retailer Program (HFSRP) was passed into law with a $250,000 appropriation (2016-2018) providing up to $25,000 in funding to small food stores for equipment to stock healthier foods and beverages. This paper describes an observational natural experiment documenting the impact of the HFSRP on store food environments, customers' purchases and diets. METHODS: Using store observations and intercept surveys from cross-sectional, convenience customer samples (1261 customers in 22 stores, 2017-2020; 499 customers in 7 HFSRP stores, and 762 customers in 15 Comparison stores), we examined differences between HFSRP and comparison stores regarding: (1) change in store-level availability, quality, and price of healthy foods/beverages; (2) change in healthfulness of observed food and beverage purchases ("bag checks"); and, (3) change in self-reported and objectively-measured (Veggie Meter®-assessed skin carotenoids) customer dietary behaviors. Differences (HFSRP vs. comparison stores) in store-level Healthy Food Supply (HFS) and Healthy Eating Index-2010 scores were assessed using repeated measure ANOVA. Intervention effects on diet were assessed using difference-in-difference models including propensity scores. RESULTS: There were improvements in store-level supply of healthier foods/beverages within 1 year of program implementation (0 vs. 1-12 month HFS scores; p = 0.055) among HFSRP stores only. Comparing 2019 to 2017 (baseline), HFSRP stores' HFS increased, but decreased in comparison stores (p = 0.031). Findings indicated a borderline significant effect of the intervention on self-reported fruit and vegetable intake (servings/day), though in the opposite direction expected, such that fruit and vegetable intake increased more among comparison store than HFSRP store customers (p = 0.05). There was no significant change in Veggie Meter®-assessed fruit and vegetable intake by customers shopping at the intervention versus comparison stores. CONCLUSIONS: Despite improvement in healthy food availability, there was a lack of apparent impact on dietary behaviors related to the HFSRP, which could be due to intervention dose or inadequate statistical power due to the serial cross-sectional study design. It may also be that individuals buy most of their food at larger stores; thus, small store interventions may have limited impact on overall eating patterns. Future healthy retail policies should consider how to increase intervention dose to include more product marketing, consumer messaging, and technical assistance for store owners.

Virtual screening-based identification of novel fatty acid synthase inhibitor and evaluation of its antiproliferative activity in breast cancer cells.

Cancer cells activate de novo lipogenesis by overexpressing the lipogenic enzymes ACLY, ACC and FASN to support rapid cell division. FASN, previously known as oncogenic antigen-519 (OA-519) catalyzes seven sequential reactions to synthesize palmitic acid (C16) from substrates acetyl CoA, and malonyl CoA. The dependence of cancer cells on FASN-derived lipids and the differential expression of FASN in cancer cells compared to their normal counterparts make it an attractive metabolic drug target in cancer therapy. In the present study, an attempt has been made to identify potent FASN inhibitors from Asinex-Synergy compound database using structure-based virtual screening. The serial docking protocols of increasing precisions identified LEG-17649942, with glide score -10.34 kcal/mol as a promising compound which can directly interact with active site residues H293 and H331. LEG-17649942 possesses drug-like pharmacokinetic properties as predicted by Qikprop. LEG-17649942 exhibited cytotoxicity in breast cancer cell lines SK-BR-3, MCF-7 and MDA-MB-231 with maximum activity against MDA-MB-231 cells with IC50 of 50 µM. The study put forward LEG-17649942 as a novel drug-lead compound against triple negative breast cancer with an exquisite binding pattern to FASN-KS domain.

Conceptual combination of disinfection in regenerative endodontics: Conventional versus laser-assisted disinfection.

BACKGROUND: Successful regeneration depends on four key elements of tissue engineering such as stem cells, growth factors, scaffold and sterile environment. Therefore, regenerative endodontic therapy requires higher degree of disinfection for successful outcomes. AIMS AND OBJECTIVE: To evaluate the clinical and radiographic outcome using conventional and laser assisted disinfection in regenerative endodontics at 1, 3 and 6 months. MATERIALS AND METHODS: 18 children with necrotic young permanent teeth were selected for the study. In Group A, in the first appointment AAE (American Association of Endodontics, Clinical Consideration for Regenerative Endodontic Procedure - 2016) disinfection protocol was employed. In Group B laser assisted disinfection (810 nm diode laser, 1 W, 20 ms Pulse length and 20 ms interval duration) was performed. Microbial samples were obtained before and after disinfection to check the level of disinfection. The cases were followed up at 1 month, 3 months and 6 months interval and evaluated for clinical outcome, periapical healing and apical response. Data were statistically analyzed with level of significance set 1% or 5% accordingly (P<0.01 or P<0.05). RESULTS: On intragroup comparison there was significant reduction in bacterial count before and after disinfection with respect to laser assisted disinfection group. (P value = 0.007) There was no significant change in the clinical outcome score in laser group at 1,3- and 6-months interval. Periapical healing score showed statistically significant results in laser group when followed from 3 months to 6 months (P value = 0.04). CONCLUSION: Along with AAE 2016 protocol, use of laser assisted disinfection resulted in improved quality of disinfection and an expeditious clinical outcome and periapical healing in necrotic young permanent teeth. Keywords: Disinfection; laser; regenerative endodontics; sodium hypochlorite; tooth apex.

A toolbox for conducting an online OSCE.

This toolbox highlights the lessons learned and the tools used to run the online OSCE at the College of Medicine and Medical Sciences, Arabian Gulf University (CMMS-AGU) using Zoom™ï¸. The examiners considered the examination to be valid in assessing all clinical skills except for psychomotor skills and students found it to be highly acceptable. We describe three phases. Planning and preparation phase in which situation analysis, aligning stakeholders, mobilizing resources, creating a shared vision, and ownership of the exam project take place. For successful implementation of examinations, detailed plans are needed including manpower, timings, number of stations and detailed description of the steps of the examination process. We provide a set of guiding questions for proper decision making related to online clinical exams. Implementation Phase in which piloting is very useful to apply improvements to the original plan and to outline the needed capacity building of the participating staff. We give a detailed description of the guiding documents, means of communication and features of ZOOM that were used. Evaluation phase we provide a guide for evaluating the process and outcome, including a list of key performance indicators.

Targeting the Id1-Kif11 Axis in Triple-Negative Breast Cancer Using Combination Therapy.

The basic helix-loop-helix (bHLH) transcription factors inhibitor of differentiation 1 (Id1) and inhibitor of differentiation 3 (Id3) (referred to as Id) have an important role in maintaining the cancer stem cell (CSC) phenotype in the triple-negative breast cancer (TNBC) subtype. In this study, we aimed to understand the molecular mechanism underlying Id control of CSC phenotype and exploit it for therapeutic purposes. We used two different TNBC tumor models marked by either Id depletion or Id1 expression in order to identify Id targets using a combinatorial analysis of RNA sequencing and microarray data. Phenotypically, Id protein depletion leads to cell cycle arrest in the G0/G1 phase, which we demonstrate is reversible. In order to understand the molecular underpinning of Id proteins on the cell cycle phenotype, we carried out a large-scale small interfering RNA (siRNA) screen of 61 putative targets identified by using genomic analysis of two Id TNBC tumor models. Kinesin Family Member 11 (Kif11) and Aurora Kinase A (Aurka), which are critical cell cycle regulators, were further validated as Id targets. Interestingly, unlike in Id depletion conditions, Kif11 and Aurka knockdown leads to a G2/M arrest, suggesting a novel Id cell cycle mechanism, which we will explore in further studies. Therapeutic targeting of Kif11 to block the Id1-Kif11 axis was carried out using small molecular inhibitor ispinesib. We finally leveraged our findings to target the Id/Kif11 pathway using the small molecule inhibitor ispinesib in the Id+ CSC results combined with chemotherapy for better response in TNBC subtypes. This work opens up exciting new possibilities of targeting Id targets such as Kif11 in the TNBC subtype, which is currently refractory to chemotherapy. Targeting the Id1-Kif11 molecular pathway in the Id1+ CSCs in combination with chemotherapy and small molecular inhibitor results in more effective debulking of TNBC.