African American (AA) women tend to experience far worse breast cancer outcomes compared to White (W) women. While the overall incidence of breast cancer is lower in AA women, the mortality rate is significantly higher. This disparity is believed to be due to a combination of genetic and epigenetic differences in the tumor and its microenvironment that contribute to more aggressive disease phenotypes, as well as socioeconomic factors, which together yield poorer prognoses for AA women. Not only are there significant differences in mortality rates due to biologic factors, but there are also differences in treatment experiences and adverse effects (AEs) associated with breast cancer therapies that also contribute to decreased survival. Previous studies on adverse events are limited, suffer from small sample sizes and lack of systematic approaches, resulting in failure to detect significant differences. Additionally, the impact of comorbidities, prior therapies, and other patient-specific variables on race treatment-related adverse events have not been adequately studied. To this end, in a new study published in the journal Biomedicines, led by Nabil Adam who is the co-founder & CEO of Phalcon, LLC and Professor Emeritus at Rutgers University and Robert Wieder who is Professor of Medicine at the New Jersey Medical School, and the Cancer Institute of New Jersey, Rutgers University, the investigators applied temporal association rule (TAR) mining to uncover race-based patterns in the association of specific AEs with breast cancer treatments. They used the Surveillance, Epidemiology, and End Results (SEER)–Medicare dataset, which is a comprehensive source of longitudinal data that links cancer incidence records from the National Cancer Institute’s SEER program with Medicare claims data. These data have detailed information on cancer diagnoses, treatments, and outcomes for patients aged 65 and older. In their investigations, the authors used inclusion criteria of women who have been diagnosed with breast cancer stages I-IV with no history of other malignancies by National Cancer Institute clinical trials standards, to ensure a study population that is representative of the general Medicare patient population for older adults.

To uncover the associations between treatments and adverse events, the investigators applied TAR mining using the FPGrowth algorithm, which allowed them to analyze temporal progression of treatments and the resulting adverse events. The FPGrowth algorithm can handle large and complex datasets and efficiently generates frequent pattern trees without the need for candidate generation. They categorized treatments into 46 comprehensive mechanistic categories, including chemotherapy, biotherapy, and hormone therapy drugs, and consolidated adverse events from ICD-9 codes into 18 categories, which facilitated a detailed analysis of the temporal associations between treatments and adverse events. The authors’ analysis showed significant race-based differences in the associations between specific treatments and adverse events. The administration of chemotherapy, biotherapy, and immunotherapy drugs showed different adverse events in AA patients compared to W patients.

Professors Adam and Wieder found that the venue of care played a crucial role in the type and frequency of adverse events. The authors demonstrated that specific treatment categories, such as Her2 antibodies, bisphosphonates, and pyrimidine analogs, were associated with different adverse events in AA and W patients in different treatment venues. For example, Her2 antibodies were more likely to be associated with anemia and neutropenia in AA patients in institutional settings, while in W patients they were linked to nausea and respiratory symptoms. In addition, AA patients treated in institutional outpatient settings had higher incidences of severe adverse events of pulmonary embolism and severe neutropenia compared to those treated in private practice settings. In contrast, W patients showed a more uniform distribution of adverse events across different care venues which meant that the healthcare setting impacted AA patients more profoundly.

The researchers stratified the data by cancer stage into early-stage (I-III) and late-stage (IV) categories. They found that early-stage (I-III) AA patients experienced higher rates of adverse events such as severe neutropenia and thrombophilia when treated with taxanes and anthracyclines compared to their W counterparts while for the late-stage (IV) patients, they found even more pronounced disparities, with AA patients having much higher rates of adverse events (severe anemia and respiratory complications). Furthermore, the authors compared the predicted treatment/adverse events associations with actual clinical data to validate the TAR mining approach and showed that there was a high degree of overlap between the predicted and actual treatment/AE associations, which confirmed the accuracy and relevance of the mined rules. For instance, the predicted associations for taxanes and Her2 antibodies matched the actual observed adverse events (nausea, neutropenia, and respiratory symptoms). In conclusion, the research work of Professors Nabil Adam and Robert Wieder uncovered temporally relevant patterns of treatment-related adverse events that were previously difficult to detect. Their use of TAR mining identified specific treatment-adverse event associations that vary by race, stage of disease, and venue of care. These findings will be of high value to clinicians who can use the authors’ data to better stratify patients based on their risk of severe adverse events. For example, the knowledge that AA patients are more likely to experience severe neutropenia with certain chemotherapies allows for closer proactive monitoring and management of these patients. Moreover, oncologists can use the findings to better communicate with patients about the potential risks associated with their treatment plans which can help patients be more informed about their care. Additionally, the study provides evidence that can be used to advocate for changes in clinical practice guidelines and healthcare policies to address racial disparities in breast cancer treatment such as recommendations for more intensive monitoring of AA patients or adjustments to standard treatment protocols based on patient demographics and therefore ensure that high-risk populations receive the support and intervention they need to manage adverse events.

Cancer remains a global health concern, and its impact extends far beyond geographical boundaries. According to the World Health Organization (WHO), cancer is the second leading cause of death worldwide, responsible for nearly 10 million deaths in 2020. It is estimated that one in six deaths globally is due to cancer. The escalating cancer incidence is a grave concern, demanding innovative solutions from the scientific community. Pharmaceutical chemists have tirelessly pursued the development of anticancer agents, resulting in a plethora of promising compounds. However, substantial challenges, including low selectivity, adverse toxic side effects, and the emergence of multidrug resistance, have proven formidable obstacles to overcome. One interesting natural product named matrine, the principal component of Sophora flavescens Aiton, has demonstrated its ability to inhibit tumor cell proliferation. Nevertheless, its clinical application remains hindered by limitations such as low water solubility, poor bioavailability, and toxic side effects. The family of dithiocarbamate (DTC)-containing molecules, exemplified by brassinin, has exhibited diverse physiological activities, providing a tantalizing avenue for anticancer drug development. Notably, chalcone-DTC, synthesized by introducing DTC into the chalcone molecule, and thalidomide-DTC, created by integrating DTC into thalidomide, have both shown increased inhibitory activity against cancer cells.

Nature has provided invaluable source of therapeutic agents, with more than 80 of the 371 pharmaceutical monographs in the Ninth Edition of the International Pharmacopoeia derived from natural products and their derivatives. Astonishingly, over 60% of existing anticancer agents are derived from natural product derivatives. Consequently, natural products present an intriguing resource for the development of novel therapies for a wide spectrum of diseases, including cancer. The amalgamation of distinct biologically active pharmacophores through the “molecular hybridization strategy” represents a potent approach for crafting highly effective antitumor drug candidates. In light of this, the fusion of DTC with matrine holds promise for the creation of derivatives endowed with exceptional antitumor activity. To this end, a new study published in the European Journal of Medicinal Chemistry conducted by Dr. Meng-Wei Zhang, Dr. Yu He, and Dr. Meng-Xue Wei from the College of Chemistry and Chemical Engineering at Ningxia University developed a one-pot, three-step synthesis strategy to generate a series of matrine-DTC hybrids and rigorously assessed their in vitro cytotoxic activity and mode of action.

The research team synthetic strategy began with the hydrolysis of matrine in aqueous potassium hydroxide, yielding product 1. This compound was subsequently esterified using a mixture of sulphoxide chloride and a primary alcohol (methanol or ethanol) to generate crude products 2. A “one-pot reaction” with carbon disulphide and halogenated hydrocarbons in chloroform followed, yielding matrine-DTC hybrids 4a–4f with an overall yield ranging from 33% to 58% over two steps. Unfortunately, replacing the primary alcohol with a secondary alcohol (L-menthol or isoborneol) in the above procedure yielded no products, likely attributable to the weak reactivity of secondary alcohols. To surmount this challenge, the authors first reacted product 1 with carbon disulphide, halogenated hydrocarbons, and potassium phosphate in chloroform to yield intermediates 3. These intermediates were subsequently condensed with L-menthol or isoborneol to generate matrine-DTC hybrids 4g–4l through a two-step process, yielding 34% to 48% overall. It is noteworthy to mention, the existence of matrine-DTC hybrids had not been previously reported, underscoring the novelty of the new study. The structures of all matrine-DTC hybrids were meticulously confirmed through ¹H, ¹³C NMR and ¹⁹F NMR spectroscopy, HRMS, and FT-IR spectroscopy.

The authors systematically evaluated the in vitro cytotoxicity of the synthesized matrine-DTC hybrids (4a–4l) and compared to both matrine and vincristine (VCR), which served as references. Remarkably, all matrine-DTC hybrids exhibited significantly greater toxicity towards human hepatoma cells HepG2 when compared to the parent matrine (IC50 > 4900 μM). Among these hybrids, 4l emerged as the most potent, with an IC50 value of 31.39 μM against HepG2 cells and the highest selectivity (SI = HEK-293T/HepG2 ≈ 6), surpassing that of VCR (SI ≈ 1) and matrine (SI ≈ 1). Notably, when R1 was menthyl or bornyl, the hybrids (4g–4l) displayed superior toxicity (IC50 = 31.39–90.68 μM) compared to when R1 was methyl or ethyl (4a–4f, IC50 = 147.78–262.45 μM) against human hepatoma cells. This enhanced potency was also superior to that of the reference VCR (IC50 = 93.67 μM). Additionally, the hybrids 4f and 4l, containing 4-(trifluoromethyl)benzyl as R2, demonstrated the highest selectivity (SI ≈ 6), further outperforming matrine and VCR.

The superiority of hybrid 4l was underscored by its remarkable cytotoxicity against various other human cancer cells, including lung cancer cells (Calu-1), breast cancer cells (SK-BR-3), liver cancer cells (HUH-7), renal cell carcinoma cells (786-O), and ovarian cancer cells (SK-OV-3). Notably, this heightened toxicity was accompanied by relatively lower toxicity towards corresponding normal cells (WI-38, LX-2, HEK-293T, and KGN), further substantiating the potential of hybrid 4l as an anti-hepatocellular carcinoma agent. To elucidate the mechanism underlying the observed cytotoxicity, the authors conducted a series of experiments. As the concentration of hybrid 4l increased, there was a corresponding rise in the inhibition rate against HepG2 cells, as well as observable changes in cell morphology, characterized by volume reduction, chromatin margination, and the formation of apoptotic vesicles. Flow cytometry using Annexin V-FITC/PI confirmed the induction of apoptosis by hybrid 4l in HepG2 cells in a concentration-dependent manner.

In summary, Ningxia University scientists successfully synthesized twelve novel matrine-DTC hybrids through a concise three-step synthetic strategy. These hybrids exhibited remarkable in vitro cytotoxicity against human hepatoma cells, with hybrid 4l standing out as the most potent candidate, outperforming both matrine and VCR. Moreover, hybrids 4f and 4l displayed exceptional selectivity, making them highly promising candidates for further exploration as anti-hepatocellular carcinoma drugs. The new study underscores the potential of molecular hybridization as a viable strategy for creating novel anticancer agents and offers new hope in the ongoing battle against cancer.

Pancreatic cancer (PaCa) ranks as the 10^th most common cancer and the 5^th leading cause of cancer mortality in the United Kingdom. The prognosis for PaCa is poor, with a five-year survival rate of only 7%. This poor survival rate is largely attributed to the typically late stage at which the disease is diagnosed, because of its asymptomatic nature in early stages and the absence of effective screening programs with significantly limited treatment options. Identifying individuals at high risk for PaCa is essential for developing early prevention strategies and improving outcomes. However, the challenge lies in the multiple PaCa risk factors, which include genetic predisposition, lifestyle factors (such as smoking and alcohol consumption), and medical history-related conditions (like diabetes mellitus and pancreatitis). Single-nucleotide polymorphisms (SNPs) have been increasingly recognized as significant contributors to cancer risk, and numerous SNPs associated with PaCa have been identified through genome-wide association studies (GWAS) which facilitated the development of polygenic risk scores (PRS) that can aggregate the effects of multiple SNPs and stratify individuals based on their genetic susceptibility to PaCa. Despite these advancements, there is still a lack of comprehensive risk prediction models that integrate the full spectrum of PaCa risk factors, encompassing genetic, lifestyle, and medical history-related variables. Most existing models focus on a limited set of risk factors and fail to capture the complex interplay of causes that may contribute to PaCa risk. This gap highlights the need for an integrated approach that can effectively identify high-risk individuals and inform targeted prevention efforts. To this end, new study published in Biomedicines and conducted by PhD candidate Te-Min Ke, Dr. Artitaya Lophatananon, and Professor Kenneth Muir from the University of Manchester developed a new integrated PaCa risk prediction model. The team performed a nested case-control study using the UK Biobank cohort, which includes comprehensive health and genetic data from over 500,000 participants with 1,402 incident pancreatic cancer cases identified after study enrollment and 257,348 cancer-free controls. Afterward, they classified the exposure variables into three categories: non-modifiable variables (gender, age, blood type, family history of bowel cancer, and PRS), lifestyle-related modifiable (tobacco smoking, alcohol intake, BMI, waist-to-hip ratio, and physical activity), and medical history-related variables (pancreatitis, diabetes mellitus, hepatitis B, gallbladder-related diseases, Helicobacter pylori infection, peritonitis, vitamin D deficiency, and systemic lupus erythematosus). They derived the PRS from 40 SNPs associated with pancreatic cancer that were identified through GWAS. The PRS provided a quantitative measure of genetic susceptibility to PaCa, stratifying participants into quintiles. Higher PRS quintiles were significantly associated with increased PaCa risk which emphasized the genetic component’s importance in risk prediction.

The authors employed a random forest model to identify the most influential risk factors for PaCa. The model was trained on 85% of the dataset and tested on the remaining 15% which ensured robust internal validation through 10-fold cross-validation. The optimal parameters for the random forest model were determined using RandomizedSearchCV and GridSearchCV functions in the Scikit-learn package. The model revealed that the top five influential features were age, PRS, pancreatitis, DM, and smoking. Other significant variables included alcohol consumption, gallbladder-related diseases, BMI, physical activity, and gender. The researchers developed also a multivariable logistic regression model to complement the random forest model using stepwise selection methods which quantified the odds ratios (ORs) for each risk factor and provided a clear interpretation of their contributions to PaCa risk. The logistic regression model identified nine significant risk factors: male gender (OR = 1.17), age (OR = 1.10 per year), non-O blood type (OR = 1.29), higher PRS quintile (Q5 vs. Q1, OR = 2.03), current smoking (OR = 1.82), higher alcohol consumption (OR = 1.27), pancreatitis (OR = 3.99), DM (OR = 2.57), and gallbladder-related diseases (OR = 2.07). Moreover, the authors created visual nomograms based on the logistic regression model which made the findings accessible and actionable and allowed users to calculate the probability of developing PaCa by summing weighted point values for each risk factor. Additionally, they developed dynamic, web-based nomogram to provide an interactive tool for immediate risk assessment in clinical and community settings. The nomogram visualization highlighted the relative importance of each risk factor, with age, pancreatitis, DM, and PRS being the most influential.  The online availability of the dynamic further enhanced the model’s usability and enabled healthcare providers and individuals to easily assess PaCa risk and implement targeted prevention strategies. In conclusion, the authors’ approach of combining the results from both models provided a comprehensive understanding of PaCa risk factors where the random forest model identified the most critical risk variables and the logistic regression model quantified their impacts. Such dual model approach ensured a robust risk prediction framework capable of integrating genetic predisposition, lifestyle factors, and medical history. Moreover, the new dynamic nomograms allow for personalized risk assessment, making it easier for healthcare providers to tailor prevention and early detection strategies to individual patients which can potentially lead to earlier diagnosis and better outcomes for patients at high risk of PaCa. Furthermore, the visual and dynamic nomograms provide an intuitive tool for clinicians to assess and easily communicate risk to patients which can enhance patient understanding and engagement.

Acknowledgment

The research leading to the results presented in this paper has received funding from the European Union’s funded Project iHELP under grant agreement no 10101744. The iHELP Project focuses on developing and utilizing AI-driven learning and decision-support technology to identify and mitigate risks associated with pancreatic cancer at an early stage. For more information about the iHelp Project, please visit their website: https://ihelp-project.eu/.

Point-of-care testing (POCT) is a type of diagnostic testing that is performed outside of the traditional laboratory setting, often at or near the site of patient care. POCT can provide rapid, real-time cost-effective results that can be used to guide clinical decision-making and improve patient outcomes. POCT provides results quickly, often within minutes, allowing for timely intervention and treatment. This is particularly important in emergency or critical care settings, where rapid diagnosis and treatment can be life-saving. Moreover, POCT can help healthcare providers make more informed clinical decisions, leading to improved patient outcomes. For example, rapid diagnosis of a bacterial infection can lead to prompt administration of antibiotics, reducing the risk of complications and improving the patient’s prognosis. Furthermore, POCT allows for testing to be performed at or near the point of care, reducing the need for transportation of specimens to a central laboratory and minimizing the time between specimen collection and test results. It can also be used in settings where traditional laboratory testing may not be available or feasible, such as in remote or resource-limited areas. The technologies used for POCT have seen significant advancements in recent years, with the development of microfluidic devices that use paper or paper-like materials to perform diagnostic tests. These devices are particularly useful in low-resource settings, where access to specialized equipment and trained personnel is limited. However, with the advent of microfluidics in POCT, alternative detection formats are required. This is where fluorescence polarization immunoassay (FPIA) comes into play. FPIA is a widely used technique for measuring the levels of various analytes in a sample. It works by measuring the degree of polarization of a fluorescent signal, which is inversely proportional to the amount of analyte in the sample. The traditional method of performing FPIA is in micro-cuvettes, such as microtiter plates.

In a new research published in the peer-reviewed journal Analytica Chimica Acta, scientists from the School of Engineering of the University of Applied Sciences Western Switzerland: Dr. E.-Diana Bojescu, Dipl.-Ing. Denis Prim, Dr. Marc E. Pfeifer and Dr. Jean-Manuel Segura, developed a new method for measuring the levels of the antibiotic tobramycin in a patient’s blood using fluorescence polarization immunoassay (FPIA) which was performed within small chambers made of glass fibers.

The authors selected tobramycin to demonstrate the power of their new technology. Tobramycin is an antibiotic medication that is commonly used in medicine to treat a variety of bacterial infections. It belongs to the class of aminoglycoside antibiotics and works by inhibiting bacterial protein synthesis, ultimately leading to bacterial cell death. Monitoring the blood concentration of tobramycin is important because it is a medication that has a narrow therapeutic index, meaning that there is a small difference between the therapeutic dose and the toxic dose. Therefore, maintaining the appropriate blood concentration of tobramycin is crucial to ensure that the drug is effective at treating the bacterial infection while avoiding toxic side effects. When tobramycin is administered, it is distributed throughout the body, including in the kidneys, where it is eliminated from the body. However, in some patients, the drug may not be effectively eliminated, resulting in an accumulation of the drug in the body. This can lead to toxicity, particularly in the kidneys and ears. By monitoring the blood concentration of tobramycin, healthcare professionals can determine whether the drug is being effectively eliminated from the body and adjust the dose accordingly to avoid toxicity. Blood concentration monitoring is typically done by measuring the drug level in the blood at specific intervals after administration, usually every 24 hours.

The research team showed that FPIA can be accurately performed within glass fiber micro-chambers. The use of glass fiber micro-chambers offers several benefits over traditional assay platforms. Firstly, the small size of the chambers increases sensitivity, allowing for the detection of the analyte in minute amounts of blood. Secondly, the micro-chambers can be used to separate plasma from whole blood before quantification, which could prove to be an important asset in the development of solutions for POCT. Lastly, the glass fiber micro-chambers are easy to fabricate and can be incorporated into a small portable device, making it suitable for use in low-resource settings.

The authors demonstrated that this novel combination of glass-fiber chambers with FPIA makes it possible to develop a POCT device for therapeutic drug monitoring using a single drop of blood. The assay could be further simplified by incorporating and drying the reagents within the micro-chambers. Furthermore, the measurement imprecision is very low (CV = 1.2%), which means that the overall analytical performance could be further improved to meet the CLIA criteria and ensure adequate dosage modifications by stabilizing temperature and environmental conditions inside a dedicated FP reader and automating the measurement process. Although POCT devices usually do not reach the precision and accuracy of commercial clinical analyzers, this could be potentially compensated by performing multiple concentration measurements after the drug administration owing to the ease of handling, the rapid turnaround time and the very low blood volume requirements of the FP assay. Thereby more accurate Bayesian estimates of the area under the concentration-time curve (AUC) could be obtained and accurate dose adjustments would be ensured.

In conclusion, this study demonstrates that the use of glass fiber micro-chambers with FPIA is a promising new approach for POCT. The technique is simple, fast, and accurate, and has the potential to be used in therapeutic drug monitoring at the point of care, particularly in low-resource settings. This technique will help healthcare professionals to monitor blood concentration of tobramycin for example and detect early signs of toxicity and intervene as necessary.

Cell spheroids are crucial three-dimensional models resembling tissue structures. They closely mimic the physiological conditions found in tissues and tumors in vivo, making them indispensable in cancer research, tissue engineering, and regenerative medicine. Spheroids exhibit unique mechanical properties due to their heterogeneous structure with an outer layer of proliferating cells, a middle region of quiescent cells, and an inner necrotic core. Understanding these mechanical properties – such as elasticity, viscoelasticity, and responses to mechanical stress – provides valuable insights into cellular interactions, structural integrity, and the influence of the microenvironment on cell behavior.

Conventional methods to study these properties, like atomic force microscopy, micro-indentation, optical tweezers, and cavitation rheology, often require specialized high-cost equipment and technical expertise which are not accessible to many researchers. To this end, a recent study by Dr. Rosalia Ferraro, Professor Sergio Caserta and Professor Stefano Guido from the University of Naples Federico II in Italy, published in Advanced Materials Technologies, introduces an innovative Rheo-optical compression assay. This method leverages standard laboratory equipment commonly found in cell culture facilities. It involves applying a load to cell spheroids using microscope coverslips and capturing the resulting deformation with an optical microscope or a smartphone equipped with magnification lenses. By analyzing images to correlate applied loads with observed deformations, researchers can accurately determine key mechanical properties such as the Young modulus.

Initially, researchers validated their Rheo-optical compression assay using spherical agarose gel particles to demonstrate its feasibility before applying it to cell spheroids; the choice of agarose is related to its status as a biomimetic gel commonly utilized in literature, as it effectively mimics the mechanical properties of tissues. Gel particles were prepared by immersing droplets of hot agarose solution into silicone oil. These particles were then transferred to cell culture plates filled with silicone oil for compression tests. Using microscope coverslips as compressive loads, researchers applied sequential loads to the agarose particles and observed their deformation under a light microscope. The stress-strain outcomes obtained from these experiments at the steady-state regime showed a linear relationship, allowing the calculation of the Young modulus. Comparing these results with values obtained using conventional rheometry, researchers found the Young modulus determined by their method to be consistent, validating the accuracy and reliability of the assay.

Afterward, the researchers applied the Rheo-optical compression assay to cell spheroids derived from PANC-1 (tumor) and NIH/3T3 (non-tumor, referred as control) cell lines. Spheroids were formed using the liquid overlay technique, which involves seeding cells in wells pre-coated with non-adhesive agarose. This setup promoted cell aggregation and spheroid formation over 5-10 days, depending on the cell type. The formed spheroids were then transferred to multiwell plates filled with culture medium. Compression was applied using microscope coverslips positioned within a 3D printed frame in order to avoid asymmetric load, and the morphological response of the spheroids to varying applied stresses was observed using an inverted microscope.

Spheroids deformed progressively with increasing loads, exhibiting distinct mechanical behaviors between the two cell lines. Specifically, PANC-1 tumor spheroids showed greater deformation compared to NIH/3T3 spheroids under the same applied stress. The researchers observed that the stress-strain data followed a linear trend up to a certain strain, beyond which the response became nonlinear. By applying Hooke’s law to the linear portion of the data, the researchers calculated the Young modulus for both cell lines and found that the PANC-1 spheroids had a significantly lower Young modulus compared to NIH/3T3 spheroids, indicating that tumor cells are softer than their non-tumor counterparts. This difference measured using the new assay can be used to distinguish between cell types based on their mechanical properties.

Furthermore, the authors employed a phenomenological model developed for foams to understand the mechanical behavior of cell spheroids beyond the linear elastic regime. The model provided a good fit for data from both NIH/3T3 and PANC-1 spheroids, suggesting that the mechanical behavior of cell spheroids can be described using principles similar to those proposed to describe foams viscoelastic response.

In addition to steady-state measurements, the researchers investigated the transient deformation of cell spheroids following the application of a load. They observed time-dependent deformation behaviors, which provided valuable insights into the viscosity of the spheroids, encompassing not only elastic but also viscous characteristics. In a more recent publication, the transient deformation was also analyzed using a model-free approach. This tool enabled the derivation of viscoelastic properties directly from the time-dependent stress and strain curves of the samples.

In conclusion, Dr. Rosalia Ferraro, Professor Sergio Caserta, and Professor Stefano Guido have successfully developed the Rheo-optical compression assay, providing a straightforward and cost-effective alternative that is readily deployable in standard cell biology laboratories. This democratization of technology expands the horizons of high-throughput mechanobiology research. Specifically, by measuring the mechanical properties of tumor spheroids, this assay enables deeper insights into the mechanical aspects of cancer progression and metastasis. It also facilitates the identification of mechanical biomarkers crucial for cancer diagnosis, prognosis, and the development of new therapeutics, with potential implementation also on clinical studies.

Triple-negative breast cancer (TNBC) accounts of 20% of all breast cancer cases is a highly aggressive subtype of breast cancer characterized by the absence of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 (HER2). A unique and perplexing aspect of TNBC is its increased incidence among women in the post-partum period. While parity is generally protective against breast cancer in the long term, recent studies have found a paradoxical transient increase in breast cancer risk immediately following childbirth. This post-partum breast cancer which is often of the triple-negative subtype is associated with worse survival outcomes and a higher propensity for metastasis. However, the biological mechanisms driving this increased risk is poorly understood which complicates efforts to develop effective preventive and therapeutic strategies. Even after extensive research, it is still a challenge to find specific molecular drivers in TNBC that could serve as therapeutic targets. Moreover, it has been difficult to find genetic mutations that drives TNBC which led researchers to investigate other potential contributors to TNBC pathogenesis, including alterations in gene expression and signaling pathways that may not involve traditional oncogenic mutations. To this account, RRAS2 which is a member of the RAS-related GTPase family has emerged as a gene of interest. RRAS2 is rarely mutated not like the classical RAS oncogenes (KRAS, NRAS, HRAS), which are frequently mutated in various cancers. However, preliminary evidence suggests that its overexpression may play a significant role in breast cancer development, particularly in TNBC. Given the gaps in understanding the molecular basis of post-partum TNBC and the urgent need for new therapeutic targets, the study aimed to investigate the role of RRAS2 overexpression in breast cancer initiation and progression, as well as its potential as a biomarker for identifying high-risk patients. New study published in Molecular Cancer and conducted by Dr. Claudia Cifuentes, Clara Oeste, Isabel Fernández-Pisonero, Alejandro Hortal, Carmen García-Macías, Jeanne Hochart, Regina Rubira, Lydia Horndler, Carlos Horndler, Xosé Bustelo, and led by Professor Balbino Alarcón from  the Consejo Superior de Investigaciones Científicas de Madrid in Spain explored the hypothesis that unmutated RRAS2, when overexpressed could act as a key oncogenic driver in TNBC especially in cases associated with recent pregnancy.

The authors generated a conditional knock-in mouse model that overexpressed wild-type human RRAS2 specifically in mammary epithelial cells and observed that all female mice overexpressing RRAS2 developed TNBC, but intriguingly this development was dependent on pregnancy. Furthermore, they showed that overexpression of RRAS2 led to the formation of breast ductal adenocarcinomas which highlights RRAS2 as a potent oncogenic driver in mammary tissue under specific physiological conditions, such as those induced by pregnancy.

The team also investigated the underlying mechanisms by which RRAS2 overexpression contributes to tumorigenesis. They conducted histological analyses of the mammary glands from RRAS2-overexpressing mice at various stages of their reproductive cycle and found that during pregnancy there was a marked increase in the proliferation of alveolar buds, however, after weaning, these structures did not regress as they normally would but instead the mammary tissue exhibited sustained hyperproliferation and delayed involution. This abnormal persistence of alveolar buds and the associated hyperproliferation likely create a microenvironment conductive to tumor development. To investigate further the molecular changes associated with RRAS2 overexpression, the authors performed RNA sequencing on tumors from the RRAS2-overexpressing mice and found significant upregulation of pathways involved in cell proliferation, extracellular matrix remodeling, and survival while apoptosis pathways were downregulated. Moreover, the researchers also identified increased activity in pro-oncogenic signaling pathways including the PI3K/Akt/mTOR and Wnt/β-catenin pathways which are known to play critical roles in cancer development and progression.

In parallel with their experimental mouse model studies, the authors performed a cohort of human breast cancer sample analysis and found that RRAS2 was overexpressed in 68% of breast cancer samples with the highest expression observed in TNBC cases and in young women who had recently given birth. Further analysis of the human breast cancer samples showed that a single-nucleotide polymorphism (SNP) in the 3′ untranslated region of the RRAS2 gene known as rs8570 was associated with increased RRAS2 expression. This SNP was more frequently found in breast cancer samples compared to blood from healthy donors. In addition, the authors found a significant number of patients who had the RRAS2 gene amplified, i.e. higher number of copies of the gene, not only in the breast tumors but also in their blood. The blood in breast cancer patients is a non-tumoral tissue. The increased frequency of the SNP allele and the pre-existing amplification of the gene in blood  indicate a potential genetic predisposition to breast cancer associated with RRAS2. According to the authors, the presence of rs8570 SNP together with RRAS2 amplification supported their hypothesis that genetic factors combined with physiological changes during and after pregnancy contribute to the development of TNBC..

In conclusion, Professor Balbino Alarcón and colleagues successfully demonstrated the correlation of the RRAS2 overexpression as a key driver of TNBC post-partum and provided a new molecular target for therapeutic intervention. They suggested that targeting RRAS2 or its downstream signaling pathways such as the PI3K/Akt/mTOR and Wnt/β-catenin pathways could provide new avenues for the development of targeted therapies potentially improve patient outcome. Moreover, the authors highlighted the importance of considering physiological changes associated with pregnancy in breast cancer’s risk and development which could lead to more personalized screening strategies and early interventions for women at risk potentially improving early detection and treatment outcomes. Additionally, the identification of a specific SNP in the RRAS2 gene associated with increased expression suggests a potential genetic biomarker for breast cancer susceptibility that could be used in the future to identify individuals at higher risk specially in populations with a high prevalence of TNBC. In a statement to Medicine Innovates, Professor Alarcon said “We hope that this research will be soon translated into useful biomarkers for risk assessment and therapy guidance“.

Intrapulmonary arteriovenous anastomoses (IPAVA) are vascular conduits in the lungs that are bypasses between the arterial and venous systems, which traditionally are not considered to be significant in healthy adults at rest. However, during exercise or hypoxia, these channels can open up and impact pulmonary gas exchange and systemic oxygen delivery. Previous studies have shown that the flow through these anastomoses, and quantified as blood flow through IPAVA, increases during exercise and hypoxia but might be suppressed in hyperoxic conditions, which suggests a potential adaptive response to altered oxygen levels. Better understanding of the IPAVA regulation is essential because it may relate directly to clinical conditions such as exercise intolerance and diseases involving pulmonary shunts, such as pulmonary arteriovenous malformations. It also has implications in patients requiring supplemental oxygen, where the impact on pulmonary blood flow and gas exchange efficiency needs careful consideration. To this end, a new study published in American Journal of Physiology-Regulatory, Integrative and Comparative Physiology and conducted by Assistant professor James Davis from Indiana University School of Medicine alongside Assistant Professor Jonathan Elliott from Oregon Health & Science University, Associate Professor Joseph Duke from Northern Arizona University, Doctor Alberto Cristobal and Professor Andrew Lovering from University of Oregon, the researchers investigated the dynamics of IPAVA during varied oxygenation states in exercise conditions. They focused on the stability of saline contrast microbubbles under different oxygen conditions and how this relates to the presence of a patent foramen ovale (PFO), a common intracardiac shunt.

In their study, the team included 32 participants and split them into three groups based on the presence of a PFO where 16 participants without PFO, 8 with PFO, and 8 who exhibited late-appearing left-sided contrast but without PFO. The inclusion of individuals with and without PFO enriches the study’s applicability to a broader population. They also included both males and females, and exercise loads were adjusted accordingly (males: 250 W, females: 175 W). They participants underwent five 4-minute bouts of constant-load cycle ergometer exercise under different fractions of inspired oxygen (F_IO₂ levels: 0.21, 0.40, 0.60, 0.80, and 1.00), using a balanced Latin Squares design to control for order effects. Afterward, the researchers used transthoracic saline contrast echocardiography to assess IPAVA flow both at rest and during exercise. The precision in the measurement techniques, including constant-load cycle ergometer challenges calibrated for gender and continuous monitoring, underscores the robustness of the experimental setup. This involved the injection of saline contrast microbubbles and scoring their appearance in the left heart as a measure of shunt activity. Moreover, they developed a bubble scoring system where they scored bubbles from 0 (no microbubbles) to 5 (extensive microbubble presence), with particular attention to changes in scores across different oxygen levels.

The authors found that at lower F_IO₂ levels (0.21, 0.40, and 0.60), bubble scores were relatively unchanged and indicated active IPAVA flow during exercise.  They observed significant reductions in bubble scores at higher oxygen levels (F_IO₂ = 0.80 and 1.00), particularly in participants without PFO, which suggests a decrease in IPAVA flow under hyperoxic conditions. To investigate the effects of PFO, participants with PFO showed higher bubble scores at an F_IO₂ of 1.00 compared to those without PFO, indicating that the presence of PFO might maintain some IPAVA flow even under conditions that would typically reduce it in non-PFO individuals.

The stability of microbubbles, as indicated by their presence across increasing F_IO₂ levels, suggests that hyperoxia leads to a functional rather than a physical closure of IPAVAs. This is inferred from the consistent decrease in bubble scores without complete disappearance, indicating that while the anastomoses remain patent, their functionality or the stability of the microbubbles is compromised. According to the authors, the decrease in IPAVA flow during hyperoxia might be due to oxygen-induced vasoconstriction or other regulatory mechanisms affecting vascular tone. This mirrors mechanisms observed in systemic circulatory adaptations, like the ductus arteriosus closure after birth.

Overall, the new study by professor James Davis and colleagues demonstrated a clear influence of hyperoxia on IPAVA function during exercise, marked by a reduction in flow that is not necessarily accompanied by physical closure. The presence of a PFO modulates the response, which suggests a complex interaction between intracardiac shunts and pulmonary vascular responses. These authors’ findings enhance our understanding of pulmonary vascular physiology, especially in response to varying oxygen levels, with significant implications for clinical management of oxygen therapy in various diseases. Additionally, future studies could investigate the molecular and cellular mechanisms underlying the oxygen sensitivity of IPAVAs and the impact of chronic exposure to different oxygen levels which could improve chronic lung disease management or adaptations to high altitude.

Extracellular vesicles (EVs) are small particles released by cells into the extracellular environment. They are encased in a lipid bilayer and can range in size from 30 to 1000 nanometers. EVs are produced by a variety of cell types and can be found in many bodily fluids, including blood, urine, saliva, and cerebrospinal fluid. These vesicles carry a cargo that includes proteins, lipids, RNA, and DNA, reflecting the physiological state of their cell of origin. EVs play a significant role in cell-to-cell communication and have been implicated in various physiological and pathological processes, including immune responses, angiogenesis, and the spread of cancer. Due to their role in intercellular communication and their presence in easily accessible bodily fluids, EVs have gained significant interest as potential biomarkers for disease diagnosis, prognosis, and therapeutic monitoring. The use of EVs as biomarkers is based on their content, which can change depending on the state of the originating cell. For example, in cancer, tumor-derived EVs can carry specific proteins, DNA mutations, or RNA species that are characteristic of the tumor cells from which they originate. By analyzing the content of EVs isolated from bodily fluids, researchers can potentially identify markers indicative of disease. For clinical applications, methods for isolating and analyzing EVs are being developed and refined. Techniques such as ultracentrifugation, size-exclusion chromatography, and immunoaffinity capture are used to isolate EVs, while tools like flow cytometry, nanoparticle tracking analysis, and various molecular biology techniques are used to analyze their content. Despite the promising potential of EVs as biomarkers, there are challenges to their widespread clinical adoption. These include the need for standardized methods for EV isolation and analysis, as well as further validation of specific EV-based markers for different diseases. As research in this area continues to advance, it is likely that the use of EVs as biomarkers will become more prevalent in clinical diagnostics and personalized medicine. A new study published in Molecular Psychiatry led by Assistant Professor Lena Smirnova, Dr. Sergio Modafferi, Dr. Charlotte Schlett, Dr. Lauren M. Osborne, Dr. Jennifer Payne & Assistant Professor Dr. Sarven Sabunciyan from Johns Hopkins University, the authors addressed a critical gap in psychiatric care: the lack of non-invasive tests for monitoring brain status. Their innovative approach focused on utilizing extracellular vesicles (EVs) and their mRNA contents as potential biomarkers for inferring the state of the brain, particularly in the context of psychiatric disorders.

The researchers developed a bioinformatic approach to identify mRNAs in EVs that are specifically expressed in certain tissues, using tissue-specific gene expression profiles. This method aimed to determine the origin of EV mRNAs found in peripheral blood. The placenta was chosen as a model to validate their approach, where they successfully identified 26 placenta-specific mRNAs in EVs circulating in maternal blood during pregnancy. This confirmed that tissue-specific mRNAs could be detected in blood EVs and used to infer their tissue of origin. Applying the same bioinformatic strategy, the team identified 181 brain-specific mRNAs in EVs from maternal blood, demonstrating the feasibility of detecting brain-derived mRNAs in peripheral blood. A significant finding was the association of 13 brain-specific EV mRNAs with postpartum depression, suggesting the potential of these mRNAs as biomarkers for psychiatric conditions. To further validate their findings, the researchers compared mRNA profiles from cells and EVs within an iPSC-derived brain model, showing a correlation between cellular and extracellular mRNA levels.

The study established the feasibility of using tissue-specific gene expression to identify the origin of EV mRNAs in blood, opening new avenues for biomarker development in psychiatric care. The discovery of 181 brain-specific mRNAs in maternal blood EVs underscores the potential of these mRNAs to reflect brain activity and their implication in psychiatric disorders. The association of specific EV mRNAs with postpartum depression highlights the potential of these biomarkers in diagnosing and monitoring psychiatric conditions. The correlation found between cellular and EV mRNA levels in the brain model suggests the possibility of using peripheral blood EV mRNAs to infer changes in the brain’s transcriptional activity. This research brings to light the potential of EV mRNAs as non-invasive biomarkers for real-time monitoring of psychiatric diseases, facilitating early diagnosis and intervention. The study by Smirnova and her colleagues represents a significant advancement in psychiatric biomarker research. By harnessing the power of EV mRNAs, they have opened a new window into the brain’s state, offering promising prospects for non-invasive, real-time monitoring of psychiatric conditions. This research lays the groundwork for future studies aimed at validating and expanding the use of EV mRNAs as biomarkers in a broader range of psychiatric disorders, moving closer to the goal of precision psychiatry.

Glutamate is the primary excitatory neurotransmitter in the central nervous system and binds to postsynaptic glutamate receptors, including N-methyl-D-aspartate (NMDA) receptors. NMDA receptors play a crucial role in synaptic plasticity, which is important for learning and memory. However, in the setting of ischemia (stroke) or brain injury, overactivation of these receptors can lead to excitotoxicity, a process that damages or kills neurons. Although excitotoxicity represents an important therapeutic target to improve outcomes in patients suffering a traumatic or ischemic brain injury, prior clinical trials have been limited by the on-target adverse effects of non-selective NMDA antagonists.  NP10679 selectively inhibits the GluN2B subunit of NMDA receptor, but has no effect on GluN2A, GluN2C, or GluN2D subunits. This selectivity is beneficial because it provides effective neuroprotection and avoids the side effects associated with non-selective inhibition of all NMDA receptors regardless of subunit composition. Furthermore, extrasynaptic NMDA receptors that contain the GluN2B subunit may be involved in processes that can lead to excitotoxicity under pathological conditions, and selectively targeting this subset of receptors would mitigate those potentially harmful effects.

The unique pH-sensitive nature of NP10679 means that it becomes more active under acidic conditions, which is relevant in the context of brain injury or disease because localized regions undergoing injury develop local tissue acidosis. Thus, NP10679 can potentially be used to prevent or minimize neuronal damage caused by abnormally high concentrations of extracellular glutamate that arise in injured tissue. This is important in conditions such as stroke, traumatic brain injury, subarachnoid hemorrhage, and certain neurodegenerative diseases, in which extracellular fluid becomes acidic due to anaerobic metabolism and failure to remove CO₂. The resulting NMDA receptor-mediated excitotoxicity is a major mechanism contributing to secondary neuronal damage. The selective and pH-sensitive characteristics of NP10679 make it a promising candidate for treating various neurological conditions because it is designed to be active in regions of injury without causing strong NMDA receptor inhibition (and thus side effects) in adjacent healthy brain tissue. Ongoing research is exploring its efficacy and safety in different disease contexts, such as the treatment of acute brain injuries, epilepsy, and chronic neurodegenerative diseases, as well as prophylactic use in at-risk populations, such as individuals who suffered subarachnoid hemorrhage, an initial stroke, or patients undergoing cardiac bypass surgery. As with any drug, the safety and efficacy of NP10679 is of primary concern. Clinical trials and rigorous testing are necessary to determine its suitability for use in humans, including understanding any potential side effects or interactions with other medications.

In a new study published in the peer-reviewed journal Clinical Pharmacology in Drug Development by Drs. Robert Zaczek, Ray Dingledine, George Koszalka, Stephen Traynelis and led by Professor Daniel Laskowitz from Duke University, the authors conducted a Phase 1 clinical trial of NP10679 that implemented a comprehensive and methodical approach to evaluate this new neuroprotective agent’s pharmacokinetic profile, safety, and tolerability.

The authors divided the trial into two phases: Single Ascending Dose (SAD) and Multiple Ascending Dose (MAD). Each phase was designed to assess different aspects of the drug’s profile in healthy volunteers. For the SAD Phase, their objective was to assess the safety, tolerability, and pharmacokinetics of a single dose of NP10679 delivered by intravenous infusion. This was a randomized, double-blind, placebo-controlled single-dose, dose-escalation trial. The study included six escalating dosing cohorts, with dosages ranging from 5 mg to 200 mg. Sentinel Dosing Adaptive Design was used, where the first two subjects (1 active, 1 placebo) were dosed and observed for 48 hours before dosing the remaining subjects in the cohort. The authors reviewed safety/tolerability and pharmacokinetic data before escalating to the next dose level.

The second (MAD) phase aimed to evaluate the safety and pharmacokinetics of NP10679 on repeated dosing until a steady state blood level was reached. Similar to the SAD study, this phase was also randomized, double-blind, and placebo-controlled. The doses included 25 mg, 50 mg, and 100 mg per day. Subjects received single 75 mL IV infusions of NP10679 or placebo over 30 minutes for five consecutive days. Subjects were observed in the clinic through 48 hours post-dose on the last day, with follow-up visits thereafter. The authors assessed treatment-emergent adverse events (TEAEs), physical examinations, infusion site examinations, laboratory findings, neuropsychiatric assessments, vital signs, and subject-reported tolerability. They also monitored neurobehavioral effects associated with NMDA  antagonism, using standardized assessments, including the Hamilton Depression Rating Scale, Mini-Mental Status Examination, as well as assessments of anxiety or dissociative states. The authors coded all adverse events and tabulated them by dose level, system organ class, and preferred term. The data revealed that the most common treatment-emergent adverse event was mild somnolence, particularly on day 1 at higher dose levels, that was readily reversed by, for example, simply speaking the subject’s name. Other TEAEs such as dizziness, headache, and tremor were noted but deemed not to impact subject safety. To estimate the pharmacokinetic profile, blood samples were drawn at various intervals post-dose in both the SAD and MAD studies and liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays were used to quantify NP10679 in plasma samples. Descriptive pharmacokinetic parameters were calculated based on plasma concentrations of NP10679 using noncompartmental analysis.

In conclusion, the outcomes of the first NP10679 trial in humans are significant for several reasons. Firstly, the drug’s safety profile was remarkably favorable. Adverse effects were minimal, with only mild somnolence observed at higher doses. This finding is important, considering the well-known side effects associated with NMDA receptor antagonists, such as cognitive dysfunction and dissociative phenomena. Moreover, the pharmacokinetic data revealed a promising half-life that supports once-daily dosing, which is a crucial factor for enhancing patient compliance and therapeutic effectiveness. Notably, the study hints at NP10679’s potential as a prophylactic agent, particularly for conditions like aneurysmal subarachnoid hemorrhage (aSAH), where it could offer neuroprotection during the vulnerable period following an aneurysm rupture. Beyond its immediate clinical applications, the trial’s findings have broader implications. The success of NP10679 underscores the feasibility of developing targeted neuroprotective therapies with minimized systemic side effects. Its pH-dependent mechanism of action opens new avenues for drug development, suggesting that similar strategies could be employed in other neurological disorders. This represents an innovative approach in the field of neuroprotection, offering a targeted and potentially safer method to mitigate the harmful effects of excitotoxicity in various neurological disorders. Further research and clinical trials on NP10679 will be essential to fully understand its potential and limitations.

Cardiovascular disease develops mostly as a result of endothelial dysfunction. Endothelial regeneration may benefit greatly from the presence of circulating endothelial progenitor cells (cEPCs). Low or undetectable thyroid stimulating hormones and normal thyroid hormones are the hallmarks of subclinical thyrotoxicosis (SCT). Despite being linked to increased cardiovascular risk (CVR) and death, the course of treatment for SCT is unknown. Circulating angiogenic cells (CACs) and endothelial progenitor cells (cEPCs) have been found to be diminished in CVR-positive circumstances.

Significant knowledge gaps exist concerning the precise molecular and biochemical mechanisms governing the effects of abnormalities in thyroid function in patients and risk of cardiovascular disease. In a new study published in Frontiers in Endocrinology, Newcastle University researchers Jason Phowira, Dr. Anuradha Doddaballapur and led by Dr. Jolanta Weaver together with Sherin Bakhashab at King Abdulaziz University investigated wether SCT was linked to abnormally high levels of cEPCs or CACs that would contribute to endothelial dysfunction and ultimately elevated CVR. The authors cultured CACs from peripheral blood mononuclear cells of SCT patients and cEPCs were quantified by flow cytometry and results was compared with controls with matched ages and sex volunteers. Moreover, the research team looked in depth at the molecular mechanisms involved by assessing CAC apoptosis and eNOS expression in response to high T3 concentrations, endothelial function, plasma asymmetric dimethylarginine levels, the in-vitro paracrine function of CACs, and plasma asymmetric dimethylarginine levels.

According to the authors, SCT is linked to a decreased cEPC count and less FMD. Due to the prevalence of SCT in this group of participants, they evaluated SCT subjects in the context of cardiovascular risk factors. It was not possible to grow pro-angiogenic cells (PACs) in-vitro from the patients they studied since they used middle-aged senior participants with CVD risk factors. In order to explore the paradigm of subclinical thyrotoxicosis, PAC cultures were established from healthy persons, while researchers solely looked at CACs and cEPCs from those patients. The authors found that compared to controls, CD34+ cells were dramatically decreased in SCT. After accounting for CVD risk, the SCT state uniquely predicted the decline in cEPCs, demonstrating that the SCT state can be categorised as having an elevated CVD risk. In comparison to controls, the number of CD34+VEGFR+2 cells was considerably lower in SCT group.

Jolanta Weaver and her colleagues for the first time demonstrated the expression of thyroid hormone receptors in PACs, indicating an important function for thyroid hormones in PACs. To simulate in vivo model of SCT, they used an in vitro model of T3 high concentration. They found that eNOS expression was decreased in PACs at higher T3 levels, indicating that greater tissue T3 levels have an impact on eNOS expression in PACs. Moreover, an increase in T3 concentration promoted PAC apoptosis, which may help to explain how cEPC levels are decreased. Their, in-vitro SCT model has also demonstrated that NO may as a role in both the mobilization of cEPC from the bone marrow and endothelial dysfunction. After accounting for age differences, their study found that FMD was considerably lower in SCT individuals (9 subjects in each group) than controls.

In conclusion, decreased cEPC count and FMD are linked to SCT, supporting higher CVR in SCT. Future trials on outcomes are necessary to see whether treating subclinical hyperactive state enhances cardiovascular outcomes. Furthermore, poor vascular health is evident in patients with SCT which is consistent with an elevated risk of cardiovascular disease. Randomized controlled interventional trials are recommended to determine whether the high risk can be reduced.