Screening Techniques Research Articles

Management of cytomegalovirus infection after liver transplantation.

Cytomegalovirus (CMV) infection is one of the primary causes of morbidity and mortality following liver transplantation (LT). Based on current worldwide guidelines, the most effective strategies for avoiding post-transplant CMV infection are antiviral prophylaxis and pre-emptive treatment. CMV- IgG serology is the established technique for pretransplant screening of both donors and recipients. The clinical presentation of CMV infection and disease exhibits variability, prompting clinicians to consistently consider this possibility, particularly within the first year post-transplantation or subsequent to heightened immunosuppression. At annual symposia to discuss CMV prevention and how treatment outcomes can be improved, evidence on the incorporation of immune functional tests into clinical practice is presented, and the results of studies with new antiviral treatments are evaluated. Although there are ongoing studies on the use of letermovir and maribavir in solid organ transplantation, a consensus reflected in the guidelines has not been formed. Determining the most appropriate strategy at the individual level appears to be the key to enhancing outcomes. Although prevention strategies reduce the risk of CMV disease, the disease can still occur in up to 50% of high-risk patients. A balance between the risk of infection and disease development and the use of immunosuppressants must be considered when talking about the proper management of CMV in solid organ transplant recipients. The objective of this study was to establish a comprehensive framework for the management of CMV in patients who have had LT.

Advances in Photoacoustic Endoscopic Imaging Technology for Prostate Cancer Detection

The rapid progress in biomedical imaging technology has generated considerable interest in new non-invasive photoacoustic endoscopy imaging techniques. This emerging technology offers significant benefits, including high spectral specificity, strong tissue penetration, and real-time multidimensional high-resolution imaging capabilities, which enhance clinical diagnosis and treatment of prostate cancer. This paper delivers a thorough review of current prostate cancer screening techniques, the core principles of photoacoustic endoscopy imaging, and the latest research on its use in detecting prostate cancer. Additionally, the limitations of this technology in prostate cancer detection are discussed, and future development trends are anticipated.

Exploring Co-Amorphous Formulations Of Nevirapine: Insights From Computational, Thermal, And Solubility Analyses.

This study aimed to assess the formation of nevirapine (NVP) co-amorphs systems (CAM) with different co-formers (lamivudine-3TC, citric acid-CAc, and urea) through combined screening techniques as computational and thermal studies, solubility studies; in addition to develop and characterize suitable NVP-CAM. NVP-CAM were obtained using the quench-cooling method, and characterized by differential scanning calorimetry (DSC), X-ray diffractometry (XRD), Fourier Transform Infrared Spectroscopy (FTIR), and polarized light microscopy (PLM), in addition to in vitro dissolution in pH 6.8. The screening results indicated intermolecular interactions occurring between NVP and 3TC; NVP and CAc, where shifts in the melting temperature of NVP were verified. The presence of CAc impacted the NVP equilibrium solubility, due to hydrogen bonds. DSC thermograms evidenced the reduction and shifting of the endothermic peaks of NVP in the presence of its co-formers, suggesting partial miscibility of the compounds. Amorphization was proven by XRD and PLM assays. In vitro dissolution study exhibited a significant increase in solubility and dissolution efficiency of NVP-CAM compared to free NVP. Combined use of screening studies was useful for the development of stable and amorphous NVP-CAM, with increased NVP solubility, making CAM promising systems for combined antiretroviral therapy.

ANTICIPATING BIOSECURITY HAZARDS: DISTRIBUTION AND RECOGNITION OF BIOLOGICAL THREATS

Understanding the potential biological weapons that can be created, the technologies involved, and the entities capable of producing them is crucial for biosecurity. Equally important is comprehending the various targets a biological weapon might be aimed at, the deployment methods, and available detection strategies to counteract illegal biological threats. This chapter delves into these aspects, starting with methods of deployment. The Center for Disease Control and Prevention (CDC) categorizes disease transmission into direct and indirect modes. Direct transmission involves contact with an infected person or environmental reservoir, including droplet spread, proven highly effective in densely populated areas. Indirect transmission occurs through contaminated objects or vectors. Vehicle-based transmission involves contaminating inanimate objects like food, water, and biological products. Vector-based transmission involves organisms carrying and transmitting pathogens. Airborne transmission, receiving significant attention for its potential in biological attacks, involves pathogens suspended in the air, posing significant risks, especially in crowded areas. Biological weapons could target humans, agriculture, technology, and the environment. Attacks on humans, often the focus of biosecurity efforts, pose catastrophic outcomes. State-sponsored programs often focus on human pathogens like Bacillus anthracis and Yersinia pestis. Agroterrorism, targeting agriculture and livestock, can lead to economic losses and social instability. Attacks on technology, utilizing synthetic biology, have shown the potential to manipulate DNA to store and deploy malicious code, highlighting vulnerabilities in sequencing systems. While attacks on the environment remain theoretical, advancements in gene drives raise concerns about manipulating ecosystems. Effective biosecurity involves pre- and post-deployment screening techniques. Pre-deployment screening, particularly gene synthesis, focuses on preventing the acquisition of DNA fragments for weapon production. Current measures involve background checks and screening against pathogen lists, yet gaps exist, including inadequate international policies and the potential to hide malicious sequences within longer ones. Post-deployment detection relies on traditional molecular diagnostic methods, but synthetic pathogens may evade detection, necessitating ongoing advancements in detection techniques. Real-time sensors and biosensors offer promising avenues for quick and accurate pathogen recognition, although challenges remain in detecting novel pathogens against a natural background. In conclusion, understanding the range of biological weapons, their potential targets, deployment methods, and detection strategies is essential for effective biosecurity. Continued research and advancements in detection technologies are crucial to staying ahead of evolving biological threats, ensuring a safer and more secure future.

Stable isotope composition of long and short term carbon pools can screen drought tolerance in cassava

Leaf carbon isotope composition (δ13C) is a promising trait for assessing genotypic differences in responses to environmental stress, particularly drought. Despite its potential, comprehensive evaluations of δ13C across various carbohydrate pools and leaf positions, in comparison with other traits, are still lacking for cassava (Manihot esculenta). This study aimed to estimate the genotypic effect size and the correlation with yield for δ13C and other commonly assessed traits in cassava. We compared the δ13C values at bulk root initiation stage of fourteen genotypes grown in the wet (1450 mm annual rainfall) and dry Caribbean (950 mm annual rainfall). We focused on both non-structural (soluble sugars) as well as structural (cellulose) carbohydrate pools across multiple leaf positions. Our results revealed significant genotypic effects across nearly all pools and leaf positions, which were equal or larger than other commonly measured traits. Interestingly, correlations between isotope composition and root yield were only observed in the dry location, with upper leaves exhibiting weaker correlations across all carbohydrate pools. Notably, while soluble sugars displayed stronger correlations with yield, they also demonstrated higher susceptibility to within-field variations. Furthermore, complementary information was found when combining δ13C with other traits such as leaf retention. Combining isotope compositions across various carbohydrate pools may reveal insights into source-sink dynamics. These results underscore the potential of δ13C as a tool for improving drought tolerance in cassava and provide a basis for its integration into cassava breeding programs, when used under dry conditions. To enhance the practical utility of δ13C as a screening technique, further research is advised to validate these findings across multiple locations, growing seasons and growth stages.

Intracellular zinc protects tumours from T cell-mediated cytotoxicity

Tumour immune evasion presents a significant challenge to the effectiveness of cancer immunotherapies. Recent advances in high-throughput screening techniques have uncovered that loss of antigen presentation and cytokine signalling pathways are central mechanisms by which tumours evade T cell immunity. To uncover additional vulnerabilities in tumour cells beyond the well-recognized antigen presentation pathway, we conducted a genome-wide CRISPR/Cas9 screen to identify genes that mediate resistance to chimeric-antigen receptor (CAR)-T cells, which function independently of classical antigen presentation. Our study revealed that loss of core-binding factor subunit beta (CBFβ) enhances tumour cell resistance to T cell killing, mediated through T cell-derived TNF. Mechanistically, RNA-sequencing and elemental analyses revealed that deletion of CBFβ disrupts numerous pathways including those involved in zinc homoeostasis. Moreover, we demonstrated that modulation of cellular zinc, achieved by supplementation or chelation, significantly altered tumour cell susceptibility to TNF by regulating the levels of inhibitor of apoptosis proteins. Consistent with this, treatment of tumour cells with a membrane-permeable zinc chelator had no impact on tumour cell viability alone, but significantly increased tumour cell lysis by CD8+ T cells in a TNF-dependent but perforin-independent manner. These results underscore the crucial role of intracellular zinc in regulating tumour cell susceptibility to T cell-mediated killing, revealing a novel vulnerability in tumour cells that might be exploited for the development of future cancer immunotherapeutics.

Toxicity of Dibutyl phthalate (DBP) toward isolated human blood lymphocytes: Apoptosis initiated from intracellular calcium enhancement and mitochondrial/lysosomal cross talk

Dibutyl phthalate (DBP) is a phthalate ester with wide application in industrial products, so human exposure can happen in workplaces and environment. Conflicting results have been acquired in researches which measured the influences of phthalates contact on immune responses in laboratory animals. Nevertheless, the straight influence of DBP on human lymphocytes and entire mechanisms of its effect against these cells continue to be unexplored. The major purpose of present research was to evaluate the mechanisms which lead to the DBP toxicity on human lymphocytes using accelerated cytotoxicity mechanisms screening (ACMS) technique. Cell viability was determined following12h incubation of lymphocytes with 0.05–1 mM DBP, and mechanistic parameters were assessed after 2, 4 and 6 h of lymphocyte treatment with ½ the IC5012h (0.3 mM), the IC5012h (0.6 mM) and twice the IC5012h (1.2 mM) of DBP. The IC5012 h of a chemical/toxicant is defined as concentration that kills 50 % of cells after 12 h of exposure. The results indicate that DBP exerts toxic effects on isolated human lymphocytes, probably through mitochondrial and lysosomal damage induced by glutathione depletion and oxidative stress. In this study, suppression of cytokines (IL2, INF-gamma and TNF-alpha) production and increase in intracellular calcium were also related to DBP induced lymphocyte toxicity.

Alias‐informed model selection (AIMS) for 7 and 8 factor no‐confounding 16‐run fractional factorial designs

AbstractNonregular fractional factorial designs are a preferable alternative to regular resolution IV designs because they avoid confounding 2‐factor interactions. As a result, nonregular designs can estimate and identify a few active 2‐factor interactions. However, due to the sometimes complex alias structure of nonregular designs, standard factor screening strategies can fail to identify all active effects. We report on a screening technique that takes advantage of the alias structure of these nonregular designs. This alias‐informed‐model‐selection (AIMS) technique has been used previously for a specific 6‐factor nonregular design. We show how the AIMS technique can be applied to 7‐ and 8‐factor nonregular designs, completing the exposition of this method for all 16‐run 2‐level designs that are viable alternatives to standard Resolution IV fractional factorial designs. We compare AIMS to three other standard analysis methods for nonregular designs, stepwise regression, the lasso, and the Dantzig selector. AIMS consistently outperforms these methods in identifying the set of active factors.

Functional Modules Map of Unexplored Chemical Space: Guiding the Discovery of Giant Birefringent Materials

AbstractFinding birefringent material with giant optical anisotropy is urgent for photonic applications. However, the application of existing birefringent materials is limited by restricted transparency wavelength or optical anisotropy. The process of trial and error is time‐consuming and inefficient in terms of resources, especially when compared to the vast design spaces for materials. The immense chemical space necessitates a high‐throughput approach for giant optical anisotropy materials design. Here, a full‐applied wavelength map of birefringent materials is constructed, from conventional visible expanding to ultraviolet and deep‐ultraviolet region by high‐throughput screening methods. And highlighted 579 materials birefringence exceeds the one of available commercial birefringent materials in corresponding regions. Significantly, 54 materials possess giant birefringence above 0.5 @1064 nm. Via developing a high‐throughput screening technique for microscopic structures with given coordination number, the study further characterizes a comprehensive map of functional modules featuring birefringent activeness. According to the macroscopic and microscopic dual‐functional maps, and a novel crystal Li2(HC3N3S3)∙5H2O with birefringence difference 0.532 @546 nm is designed and synthesized successfully. This study identifies desired materials with high performance in unheeded chemical spaces.

Detection of malignant breast tissue using SAR observation with microwave imaging and convolutional neural network

The research paper introduces a deep learning approach for distinguishing between benign and malignant breast tissues using Ultra Wide Band (UWB) microwave imaging. A multi-resonant monopole microstrip patch antenna was crafted and placed on a female breast phantom model containing a tumor. The phantom’s dielectric properties were adjusted to mimic those of benign and malignant tissues. The Specific Absorption Rate (SAR) was analyzed by rotating the antenna at various angles throughout the phantom’s trajectory. Images capturing the SAR distribution over the breast phantom at different antenna resonances were obtained. The popular Convolutional Neural Network (CNN) based AlexNet model was employed for autonomous feature learning from SAR images. To address the challenge of overfitting with a limited dataset, the study utilized image augmentation and transfer learning methods. The proposed model achieved an impressive 98.59% accuracy in classifying benign and malignant breast phantom images, surpassing the performance of three other CNN models—VggNet16, GoogleNet, and ResNet. Notably, this microwave imaging system based on SAR images detected tumors as small as 1 mm with an accuracy of 97.08% outperforming existing malignancy screening techniques.

Optimized liquid chromatography-tandem mass spectrometry protocol for enhanced detection of 45 pesticides in water and soil samples.

The development of analytical screening techniques for pesticides is crucial for preventing and mitigating environmental contamination. Mass spectrometry-based screening methods differ based on the complexity of the sample matrix and the diversity of the target compounds. One of the major challenges is balancing cost reduction in the extraction process with the optimization of analytical results. This protocol introduces a universal and efficient scheme for the qualitative and quantitative schemes for 45 pesticides within a single analytical run. Water samples were extracted using an SPE column, with the pH adjusted to 7. Soil samples were processed using a modified QuEChERS method. The pretreatment for water samples emphasized selecting appropriate SPE columns and optimizing pH, while for soil samples, the focus was on choosing suitable extraction solvents and extraction salt packages. The enriched samples were then analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The method was evaluated for accuracy, precision, detection limits, and matrix effects. The method enabled the simultaneous detection of 45 pesticides within a 15-minute analysis period. SPE recoveries ranged from 56.1% to 118.8%. Instrumental detection limits varied between 0.02 and 1pg, while method detection limits extended from 0.05 to 18.47 ng/lin soil and water matrices. The approach was successfully applied to water and soil samples, with the pesticide concentration ranging from 0.1ng/L to 38 μg/L. The protocol substantially enhances the characterization and quantification of 45 pesticides in environmental samples, achieving a remarkable reduction in detection limits by an order of magnitude compared to previous research. This method enables the simultaneous detection of pesticides in both water and soil matrices using a single system, addressing the challenges of using separate systems for different environmental media. Furthermore, this protocol provides a crucial theoretical foundation for managing and safeguarding against pesticide pollution.

Advances in Understanding and Managing Gestational Diabetes Mellitus: A Comprehensive Review

Gestational Diabetes Mellitus (GDM) is a significant and growing concern in prenatal care, impacting maternal and neonatal health. This comprehensive review highlights recent advancements in the understanding and management of GDM. The pathophysiology of GDM involves complex interactions between genetic, environmental, and hormonal factors that disrupt glucose homeostasis during pregnancy. Improved screening techniques and diagnostic criteria have led to earlier and more accurate detection. Management strategies have evolved to include not only traditional lifestyle interventions such as diet and exercise but also novel pharmacological treatments and continuous glucose monitoring technologies. The role of multidisciplinary care teams in optimizing pregnancy outcomes is emphasized, ensuring tailored care that addresses the unique needs of each patient. Additionally, the review explores emerging research on the long-term implications of GDM for both mother and child, underscoring the importance of postpartum follow-up and preventive measures. These advancements collectively aim to enhance maternal and fetal outcomes, reduce complications, and provide a framework for future research and clinical practice in GDM.

Correlation of Hypothyroidism with Age and Comorbidities Among Women: A Cross-Sectional Study

Background Hypothyroidism, an endocrine disorder caused by thyroid dysfunction found to be more prevalent globally in females than in males. Young females are more prone to subclinical hypothyroidism (SCH). The study aimed to estimate the different types of hypothyroidism based on the clinical profile of thyroid hormone and to detect various comorbidities in correlation with age in women. In female patients with hypothyroidism, there is a significant association between age groups and the type of hypothyroidism (subclinical, clinical, overt), with younger age groups showing a higher prevalence of SCH. Additionally, comorbidities, such as polycystic ovarian disorder (PCOD), are more prevalent in hypothyroidism patients, particularly in those with SCH. Materials and Methods A prospective cross-sectional study executed in a tertiary care hospital for one year with an inclusion of female hypothyroid patients above 18 years of age. An informed written consent form was received from each patient with clarification of the protocol. Results A total of 44.6% women were in the 18–28 years of age group (young females) and 42.8% of women with SCH. PCOD (38.2%) was a predominant comorbid condition among hypothyroidism women. In the age group of 18–28 years, there were significantly more patients 114 (81.4%) with SCH ( p < 0.01) than those with clinical 22 (18.4%) and overt hypothyroidism 10 (14.7%). The hypothyroidism of women with PCOD 95 (67.8%) was significantly ( p < 0.01) more in subclinical than clinical 28 (23.5%) and overt hypothyroidism 2 (2.9%). Conclusion Thyroid dysfunction is uncertain in females, bulges to complications of maternal and fetal outcomes, and is associated with other comorbidities, leading to an increase in the mortality rate. Hypothyroidism is a growing burden for other comorbidities. Pharmaceutical care by healthcare professionals is to be adopted with prompt diagnostic and screening techniques.

Mycobacterial Targets for Thiourea Derivatives: Opportunities for Virtual Screening in Tuberculosis Drug Discovery.

Tuberculosis (TB) remains a primary global health concern, necessitating the discovery and development of new anti-TB drugs, mainly to combat drug-resistant strains. In this context, thiourea derivatives have emerged as promising candidates in TB drug discovery due to their diverse chemical structures and pharmacological properties. This review aimed to explore this potential, identifying and exploring molecular targets for thiourea derivatives in Mycobacterium tuberculosis (Mtb) and the potential application of virtual screening techniques in drug discovery. We have compiled a comprehensive list of possible molecular targets of thiourea derivatives in Mtb. The enzymes are primarily involved in the biosynthesis of various cell wall components, including mycolic acids, peptidoglycans, and arabinans, or targets in the branched-chain amino acid biosynthesis (BCAA) pathway and detoxification mechanisms. We discuss the potential of these targets as critical constituents for the design of novel anti-TB drugs. Besides, we highlight the opportunities that virtual screening methodologies present in identifying potential thiourea derivatives that can interact with these molecular targets. The presented findings contribute to the ongoing efforts in TB drug discovery and lay the foundation for further research in designing and developing more effective treatments against this devastating disease.

Community Pharmacists' Readiness for Breast Cancer Mammogram Promotion: A National Survey from Jordan.

Mammography is the gold standard screening technique for early detection of breast cancer. This study aimed to assess the knowledge of community pharmacists of different aspects emphasized by the JBCP programs. This study also identifies the attitudes and barriers towards promoting early detection services. This study was a cross-sectional survey of community pharmacists in Jordan. Pharmacists were randomly selected and asked to complete an electronic questionnaire. Inclusion criteria: a pharmacist with a bachelor's degree or higher and registered at the JPA working in a community pharmacy. The questionnaire included demographic and socioeconomic information, knowledge, attitudes towards breast cancer screening mammography services, and barriers towards participation in the promotion of these services. A total of 1,088 community pharmacists were approached, 1,000 (91.8%) completed the questionnaire. Participants had an average age of 34 years ± 10.8 and average experience of 9.1 ± 9.5 years. Only 48 (37.8%) of the female pharmacists aged 40 years or older underwent a mammogram. Knowledge of symptoms of breast cancer was the highest with a score of 755, followed by knowledge of risk factors (670) and finally early detection of breast cancer (540). Many barriers were reported by the community pharmacists including lack of educational materials and time constraints. Pharmacists with higher educational levels (p<0.001), of female gender (p<0.001), attended continuous cancer-related education (p<0.001), encountered a higher percentage of female customers (p<0.001), in a certain geographic location (p=0.003), underwent mammography (p=0.014), and encountered high frequency of inquiries on mammogram by the customers (p<0.001) were all associated with higher knowledge scores. Despite the reported barriers and insufficient knowledge in certain aspects of early detection of breast cancer, community pharmacists have positive attitudes and can be a valuable asset for awareness-raising efforts.

Addressing the Global Challenge: Strategies for Cervical Prevention and Early Detection within WHO's 90-70-90 Framework

Cervical cancer represents a substantial global health issue, standing as the third most prevalent cancer globally and the principal cause of cancer-associated mortality in women, with an uneven burden shouldered by less developed countries. Despite being the most preventable malignancy, with nearly all cases attributed to HPV infection, which has a lifetime cumulative risk of 80% through sexual transmission, cervical cancer remains a pressing issue. National cervical cytology screening programs have been implemented to identify cervical abnormalities, lesions, and early malignancies in eligible women. However, the utilization of screening techniques is declining, and due to resource constraints, screening schemes have not been adopted in less-developed economies. Several nations have embraced alternative screening techniques, such as Visual Inspection with Acetic Acid (VIA), which have shown impacts on mortality and morbidity rates. With the rising accessibility of efficacious HPV screening and self-testing, there will be a necessity to reconfigure existing cervical screening programs. Furthermore, preventative HPV vaccination is an integral part of the World Health Organization’s (WHO) Global Strategy to Expedite the Eradication of Cervical Cancer as a Public Health Issue, necessitating the restructuring of vaccination endeavors. This review article evaluates potential techniques for detecting precancerous lesions within the context of the WHO's 90-70-90 global policy for cervical cancer prevention and early diagnosis. The incidence and mortality rates are examined, highlighting the effectiveness of cervical cancer prevention efforts.

Policy brief Belgian EBCP mirror group ‘prevention’ and ‘early detection & screening’ in cancer

The availability and linkage of comprehensive data on cancer mortality, incidence, screening and vaccination (HPV/HB) are essential for monitoring the process and impact of all screening and vaccination programmes, and to generate evidence to improve prevention policies. Linkage of data registries will allow better exploitation of real-world data. Simplification of data sharing for epidemiological research is of utmost importance. Human, as well as technical/technological and legal/regulatory barriers need to be tackled in Belgium to aid actors involved in screening, to facilitate organised linkage of data. Rapid advances in technologies for early detection and screening for a broad range of cancer types provide significant opportunities. Such tests are not yet ready for adoption in screening programmes. Emerging evidence should be monitored to ensure that promising innovations can transition to implementation studies in a timely manner. More and sustained research is needed to generate the necessary scientific relevance for new screening techniques in relevant cancer types.

Questionnaire-Based Study on the Knowledge of Cervical Cancer and Attitude and Perception Toward Self-Sampling Method of Screening, Among Women in Sikkim, India.

Cervical cancer (CC) is the second most common cancer among women in India and in Sikkim. Screening helps in prevention and early detection of the disease; however, pelvic examination for smear collection is not preferred by many. Self-sampling method can be a good alternative for collection of smears, thus increasing screening uptake. Therefore, a study was conducted to evaluate perception about self-sampling method of screening among women in Sikkim. A total of 210 women who consented for this study were screened for cervical cancer using Pap smear and HPV DNA. Women were taught to collect their self-sample for HPV DNA test. Questionnaires were used to assess their attitude toward self-sampling prior to collection of samples and perception, post-collection. Seventy-percent women knew that CC is the most common cancer among women in Sikkim. 73% knew that it could be prevented, but only 57% had knowledge about screening facilities in Sikkim. 50% women knew about HPV vaccination program in Sikkim. 96% women had never done a self-sampling, while 67% had never heard about it. There was a significant change (p value < 0.00001) in perception toward self-sampling after the procedure. Comparative analysis showed that women found self-collection easier than expected. If they had a choice, they would prefer self-sampling over physician sampling; however, due to their trust in physicians they still believed physician collected sample would be better. Self-sampling is a socially acceptable screening technique and can increase compliance among socially shy community.

Fused in silico and bioactivity evaluation method for drug discovery: T001-10027877 was identified as an antiproliferative agent that targets EGFRT790M/C797S/L858R and EGFRT790M/L858R

BackgroundFacing the significant challenge of overcoming drug resistance in cancer treatment, particularly resistance caused by mutations in epidermal growth factor receptor (EGFR), the aim of our study was to identify potent EGFR inhibitors effective against the T790M/C797S/L858R mutant, a key player in resistance mechanisms.MethodsOur integrated in silico approach harnessed machine learning, virtual screening, and activity evaluation techniques to screen 5105 compounds from three libraries, aiming to find candidates capable of overcoming the resistance conferred by the T790M and C797S mutations within EGFR. This methodical process narrowed the search down to six promising compounds for further examination.ResultsKinase assays identified three compounds to which the T790M/C797S/L858R mutant exhibited increased sensitivity compared to the T790M/L858R mutant, highlighting the potential efficacy of these compounds against resistance mechanisms. Among them, T001-10027877 exhibited dual inhibitory effects, with IC50 values of 4.34 µM against EGFRT790M/C797S/L858R and 1.27 µM against EGFRT790M/L858R. Further investigations into the antiproliferative effects in H1975, A549, H460 and Ba/F3-EGFRL858/T790M/C797S cancer cells revealed that T001-10027877 was the most potent anticancer agent among the tested compounds. Additionally, the induction of H1975 cell apoptosis and cell cycle arrest by T001-10027877 were confirmed, elucidating its mechanism of action.ConclusionsThis study highlights the efficacy of combining computational techniques with bioactivity assessments in the quest for novel antiproliferative agents targeting complex EGFR mutations. In particular, T001-10027877 has great potential for overcoming EGFR-mediated resistance and merits further in vivo exploration. Our findings contribute valuable insights into the development of next-generation anticancer therapies, demonstrating the power of an integrated drug discovery approach.

Genome-Wide Screening Approaches for Biochemical Reactions Independent of Cell Growth.

Genome-wide screening is a potent approach for comprehensively understanding the molecular mechanisms of biological phenomena. However, despite its widespread use in the past decades across various biological targets, its application to biochemical reactions with temporal and reversible biological outputs remains a formidable challenge. To uncover the molecular machinery underlying various biochemical reactions, we have recently developed the revival screening method, which combines flow cytometry-based cell sorting with library reconstruction from collected cells. Our refinements to the traditional genome-wide screening technique have proven successful in revealing the molecular machinery of biochemical reactions of interest. In this article, we elucidate the technical basis of revival screening, focusing on its application to CRISPR-Cas9 single guide RNA (sgRNA) library and complementary DNA (cDNA) library screening. Finally, we also discuss the future of genome-wide screening while describing recent achievements from in vitro and in vivo screening.