Conventional Screening Methods Research Articles

High-throughput single-cell antibody secretion quantification and enrichment using droplet microfluidics-based FRET assay

SummaryHigh-throughput screening and enrichment of antibody-producing cells have many important applications. Herein, we present a droplet microfluidic approach for high-throughput screening and sorting of antibody-secreting cells using a Förster resonance electron transfer (FRET)-based assay. The FRET signal is mediated by the specific binding of the secreted antibody to two fluorescently labeled probes supplied within a droplet. Functional hybridoma cells expressing either membrane-bound or secreted monoclonal antibodies (mAbs), or both, were efficiently differentiated in less than 30 min. The antibody secretion rate by individual hybridoma cells was recorded in the range of 14,000 Abs/min, while the density of membrane-bound fraction was approximately 100 Abs/μm2. Combining the FRET assay with droplet-based single-cell sorting, an 800-fold enrichment of antigen-specific cells was achieved after one round of sorting. The presented system overcomes several key limitations observed in conventional FACS-based screening methods and should be applicable to assaying various other secreted proteins.

Acceptability of Human Papilloma Virus Self-Sampling for Cervical Cancer Screening in a Cohort of Patients from Romania (Stage 2).

(1) Background: Low patient’s adherence to conventional cervical cancer screening methods determined the need to take into consideration alternative approaches, and vaginal HPV self-sampling is one of them. We aimed to evaluate, using an online survey, the Romanian women’s acceptability of vaginal HPV self-sampling. (2) Methods: A 13-questions online survey was distributed on three Facebook groups, and the results were summarized. (3) Results: Despite of good educational background, 10.8% (n = 60) of the respondents did not know what a Pap smear is, and 33% (n = 183) were not informed about the free national cervical cancer screening program. Multivariate analysis revealed an increased likelihood of vaginal self-sampling acceptance among respondents who did not know about Pap test (OR: 7.80; 95%CI: 1.062–57.431; p = 0.021), national cervical cancer screening program (OR: 1.96; 95%CI: 1.010–3.806; p = 0.02), HPV infection (OR: 7.35; 95%CI: 3.099–17.449; p< 0.001) or HPV test (OR: 1.67; 95%CI: 0.950–2.948; p = 0.03). Moreover, women who did not previously undergo a cervical cancer screening program were more likely to accept the new screening method (OR: 1.62; 95%CI: 0.878–3.015; p = 0.04). (4) Conclusions: Our results showed high acceptability rates of vaginal HPV self-sampling among participants.

Health Supervision for Children and Adolescents With Down Syndrome.

Health Supervision for Children and Adolescents With Down Syndrome.

In silico Drug Screening Approach Using L1000-Based Connectivity Map and Its Application to COVID-19.

Conventional drug screening methods search for a limited number of small molecules that directly interact with the target protein. This process can be slow, cumbersome and has driven the need for developing new drug screening approaches to counter rapidly emerging diseases such as COVID-19. We propose a pipeline for drug repurposing combining in silico drug candidate identification followed by in vitro characterization of these candidates. We first identified a gene target of interest, the entry receptor for the SARS-CoV-2 virus, angiotensin converting enzyme 2 (ACE2). Next, we employed a gene expression profile database, L1000-based Connectivity Map to query gene expression patterns in lung epithelial cells, which act as the primary site of SARS-CoV-2 infection. Using gene expression profiles from 5 different lung epithelial cell lines, we computationally identified 17 small molecules that were predicted to decrease ACE2 expression. We further performed a streamlined validation in the normal human epithelial cell line BEAS-2B to demonstrate that these compounds can indeed decrease ACE2 surface expression and to profile cell health and viability upon drug treatment. This proposed pipeline combining in silico drug compound identification and in vitro expression and viability characterization in relevant cell types can aid in the repurposing of FDA-approved drugs to combat rapidly emerging diseases.

HPV and DNA Methylation Testing in Urine for Cervical Intraepithelial Neoplasia and Cervical Cancer Detection.

Biomarker detection in urine offers a potential solution to increase effectiveness of cervical cancer screening programs by attracting nonresponders. In this prospective study, the presence of high-risk human papillomavirus (hrHPV) DNA and the performance of DNA methylation analysis was determined for the detection of cervical cancer and high-grade cervical intraepithelial neoplasia (CIN2/3) in urine, and compared with paired cervicovaginal self-samples and clinician-taken cervical scrapes. A total of 587 samples were included from 113 women with cervical cancer, 92 women with CIN2/3, and 64 controls. Samples were tested for hrHPV DNA and five methylation markers. Univariate and multivariate logistic regression and leave-one-out cross-validation were used to determine the methylation marker performance for CIN3 and cervical cancer (CIN3+) detection in urine. Agreement between samples was determined using Cohen kappa statistics and the Spearman correlation coefficients. HrHPV presence was high in all sample types, 79% to 92%. Methylation levels of all markers in urine significantly increased with increasing severity of disease. The optimal marker panel (ASCL1/LHX8) resulted in an AUC of 0.84 for CIN3+ detection in urine, corresponding to an 86% sensitivity at a 70% predefined specificity. At this threshold 96% (109/113) of cervical cancers, 68% (46/64) of CIN3, and 58% (14/24) of CIN2 were detected. Between paired samples, a strong agreement for HPV16/18 genotyping and a fair to strong correlation for methylation was found. HrHPV DNA and DNA methylation testing in urine offers a promising solution to detect cervical cancer and CIN2/3 lesions, especially for women currently unreached by conventional screening methods.

OP015: Multi-omic analysis of circulating tumour DNA for the early detection of cancer in patients with Li-Fraumeni syndrome

Patients with Li-Fraumeni Syndrome (LFS) harbor germline pathogenic variants in the TP53 gene causing predisposition to many cancers from childhood through late adulthood and can accelerate the onset of cancer by >30 years. The management of LFS patients involves regular surveillance including MRI of the brain, breast, and whole body. This aggressive clinical surveillance for early tumour detection is associated with improved survival but is disruptive to quality of life. For some cancers, conventional screening methods may not be entirely effective.

Host status of Bambara groundnut accessions to Meloidogyne incognita using conventional screenhouse and novel seedling pouch methods

Summary Bambara groundnut (Vigna subterranea) is an underexploited legume indigenous to sub-Saharan Africa with high protein content. Production constraints of Bambara groundnut include the root-knot nematodes (Meloidogyne spp.), which causes substantial losses that present an additional burden that growers need to manage. In this study we: i) assessed genetic resistance against Meloidogyne incognita in Bambara groundnut; and ii) evaluated an improved, more efficient screening method for Bambara. Fifty accessions were evaluated in two screenhouse trials. Following this, the use of seedling pouches was assessed for a high throughput screening method, using two accessions identified from each of three host status categories (resistant, tolerant and susceptible). Variability in host response was observed between the trials but five accessions demonstrated good resistance to M. incognita. A general reduction in plant growth parameters was observed on inoculated plants, especially for chlorophyll content at 11 weeks after planting. The pouch screening system confirmed the screenhouse pot results but appears to be more sensitive. From a range of inocula no differences in gall index were observed over the 28 days of assessment in pouches but the number of egg masses proved a useful parameter and differed between inoculum levels, with 500 juveniles/eggs determined as the optimum inoculum level. The use of seedling pouches reduced time, space and cost to screen Bambara groundnuts for resistance against M. incognita. Consequently, it is recommended as an efficient, high throughput, rapid screening protocol for Bambara groundnut compared to conventional pot screening methods.

Automated Segmentation of Mass Regions in DBT Images Using a Dilated DCNN Approach.

To overcome the limitations of conventional breast screening methods based on digital mammography, a quasi-3D imaging technique, digital breast tomosynthesis (DBT) has been developed in the field of breast cancer screening in recent years. In this work, a computer-aided architecture for mass regions segmentation in DBT images using a dilated deep convolutional neural network (DCNN) is developed. First, to improve the low contrast of breast tumour candidate regions and depress the background tissue noise in the DBT image effectively, the constraint matrix is established after top-hat transformation and multiplied with the DBT image. Second, input image patches are generated, and the data augmentation technique is performed to create the training data set for training a dilated DCNN architecture. Then the mass regions in DBT images are preliminarily segmented; each pixel is divided into two different kinds of labels. Finally, the postprocessing procedure removes all false-positives regions with less than 50 voxels. The final segmentation results are obtained by smoothing the boundaries of the mass regions with a median filter. The testing accuracy (ACC), sensitivity (SEN), and the area under the receiver operating curve (AUC) are adopted as the evaluation metrics, and the ACC, SEN, as well as AUC are 86.3%, 85.6%, and 0.852 for segmenting the mass regions in DBT images on the entire data set, respectively. The experimental results indicate that our proposed approach achieves promising results compared with other classical CAD-based frameworks.

Second-look Surgery Detects Low Volume Recurrent Disease Following Cytoreductive Surgery for Peritoneal Carcinomatosis.

Formation of stoma during cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) for peritoneal carcinomatosis (PC) is often performed to reduce the risk of anastomotic leak. Subsequent stoma reversal provides a unique opportunity for second-look surgery to detect early peritoneal recurrence. Current surveillance methods often fail to detect disease early, including imaging and biochemical markers. In our study, we examined the safety and efficacy of second-look surgery for detection and treatment of disease recurrence. We performed a retrospective analysis of prospectively collected data from 35 patients undergoing stoma reversal from 2015 to 2019 with negative pre-operative imaging. A total of 37% of cases had disease recurrence, with a median peritoneal cancer index of 4. Complete cytoreduction was achieved in all patients. The majority of patients (77%) suffered minor complications only. Median length of hospital stay was 12 days. Second-look surgery detects early disease recurrence and is a safe alternative to conventional screening methods post primary CRS/HIPEC for PC. Long-term, routine second-look surgery can improve survival.

RNA-Seq of Single Fish Cells - Seeking Out the Leukocytes Mediating Immunity in Teleost Fishes.

The immune system is a complex and sophisticated biological system, spanning multiple levels of complexity, from the molecular level to that of tissue. Our current understanding of its function and complexity, of the heterogeneity of leukocytes, is a result of decades of concentrated efforts to delineate cellular markers using conventional methods of antibody screening and antigen identification. In mammalian models, this led to in-depth understanding of individual leukocyte subsets, their phenotypes, and their roles in health and disease. The field was further propelled forward by the development of single-cell (sc) RNA-seq technologies, offering an even broader and more integrated view of how cells work together to generate a particular response. Consequently, the adoption of scRNA-seq revealed the unexpected plasticity and heterogeneity of leukocyte populations and shifted several long-standing paradigms of immunology. This review article highlights the unprecedented opportunities offered by scRNA-seq technology to unveil the individual contributions of leukocyte subsets and their crosstalk in generating the overall immune responses in bony fishes. Single-cell transcriptomics allow identifying unseen relationships, and formulating novel hypotheses tailored for teleost species, without the need to rely on the limited number of fish-specific antibodies and pre-selected markers. Several recent studies on single-cell transcriptomes of fish have already identified previously unnoticed expression signatures and provided astonishing insights into the diversity of teleost leukocytes and the evolution of vertebrate immunity. Without a doubt, scRNA-seq in tandem with bioinformatics tools and state-of-the-art methods, will facilitate studying the teleost immune system by not only defining key markers, but also teaching us about lymphoid tissue organization, development/differentiation, cell-cell interactions, antigen receptor repertoires, states of health and disease, all across time and space in fishes. These advances will invite more researchers to develop the tools necessary to explore the immunology of fishes, which remain non-conventional animal models from which we have much to learn.

Methods of screening, monitoring and management of cardiac toxicity induced by chemotherapeutics

Cardiac toxicity is one of the most common side effects of anticancer drugs. Cardiac toxicity results dysfunction of heart including hypotension, heart failure, and even cause death in extreme cases. The potential risk of cardiotoxicity is a huge concern in chemotherapeutics mediated cancer treatment. The individual with any pre-existing cardiac issues are excluded from clinical trials due to the potential risk of cardiotoxicity. Because of the potential cardiotoxicity, there is an emerging need for alternatives of some very potent anticancer drugs (doxorubicin/DOX, 5-fluorouracil/5FU, trastuzumab). While a patient is being treated with anticancer drugs, early blood screening, biomarker detection, and careful monitoring of cardiac functions are necessary to be able to avoid any irreversible cardiac damage. Therefore, early detection methodology to monitor cardiotoxicity in real-time, and a drug formulation that prevent interaction between drug and cardiac cell, seemingly have potential to mitigate the risk. In this review, we have summarized the cardiotoxicity of the most used anticancer drugs, their pathophysiology and some of the conventional and newer screening methods available to manage an individual patient in clinic. We have also incorporated our perspective on how a rationale designing of biomolecules can be used to overcome the cardiotoxicity generated chemotherapeutics.

Biomarkers of coordinate metabolic reprogramming and the construction of a co-expression network in colorectal cancer.

Globally, the incidence and mortality of colorectal cancer (CRC) rank amongst the highest of all malignancies. Thus, research aimed at developing new screening strategies and biomarkers for the early detection of CRC is needed. At present, conventional screening methods have limitations; therefore, new testing strategies have been considered. Using metabolomics to explore the molecular changes in CRC tissue is a mainstream method for identifying potential biomarkers and key cancer factors. In the present study, 27 samples from nine CRC patients were used to analyze the metabolite differences between the tumor, paracancerous, and normal tissues. The metabolite differences in the various stages of CRC (stages IIA, IIB, and IIIC) were analyzed as well. Subsequently, principal component analysis (PCA), permutation, and trend analyses were performed. Weighted gene co-expression and metabolite-metabolite interaction networks were also constructed. A total of 5,834 metabolites were identified among the included samples. Permutation analysis showed a clear separation between the different tissues and different stages. Compared with normal tissues, tumor tissues exhibited 11, 233, and 25 up-regulated metabolites as well as one, 77, and zero down-regulated metabolites in stages IIA, IIB, and IIIC, respectively. Moreover, tumor tissues in stage IIB exhibited more differential metabolites (233 up-regulated and 77 down-regulated). Weighted Gene Correlation Network Analysis (WGCNA) clustered the 5,834 metabolites into 15 different modules, of which four modules were significantly correlated with tissue specificity. Notably, glycerophospholipid metabolism, fatty acid metabolism, and other pathways were enriched in these modules. Fatty acids and glycerophospholipids were significantly related to the development of CRC. This result is of great significance for future targeted screening of CRC biomarkers and further clarifying the nutrient metabolism of cancer cells.

A Novel Miniaturized mmWave Antenna Sensor for Breast Tumor Detection and 5G Communication

Breast cancer is one of the deadliest forms of disease that affects women worldwide. X-ray mammography, magnetic resonance imaging, and ultrasound are all helpful methods of medical imaging, but they have their own limitations due to low tissue contrast and side effects. Millimeter wave (mmWave) imaging has been offered as a viable alternative to the conventional screening methods due to their drawbacks. Without exposing the patient to ionizing radiations, malignant lesions can be detected by using the dielectric discontinuities in the tissues. The purpose of this work is to develop a simple, low-cost miniaturized mmWave imaging antenna sensor that can be used to detection of breast tumors or cancers in women by monitoring the changes in the antenna’s S11 parameter. The dimension of the proposed rectangular mmWave antenna is 5 mm × 5 mm × 0.578 mm. The proposed antenna sensor has an operating frequency band range of 32.626 GHz to 33.96 GHz with a peak gain of 6.65 dB and an efficiency of 91.46% in terms of radiation which makes it a suitable system for fifth generation (5G) communication. As well as the antenna sensor can detect very tiny malignant that size ≥1 mm inside the breast fantom.

EVALUATION OF THE VALIDITY OF HEMOCUE HB 201+ FOR PREDONATION ASSESSMENT OF LOW AND HIGH HAEMOGLOBIN LEVELS IN BLOOD DONORS: ONE YEAR PROSPECTIVE STUDY IN BLOOD BANK OF GMC DODA

Introduction: Predonation assessment of haemoglobin in blood donors is important to avoid unnecessary rejection of potential blood donors. Compared to the conventional screening methods for haemoglobin assessment the method to be used in blood banks needs to be rapid, sensitive and accurate for both low and high haemoglobin values. Materials and methods: This was a prospective study conducted in blood bank of GMC Doda w.e.f 1st April 2020 to 31st March 2021. Finger prick samples were taken from 1214 potential blood donors and hemoglobin was estimated using HemoCue Hb 201+. Also venous samples of potential blood donors were obtained and run on automated haematologyanalysers. The values were compared. Male donors with haemoglobin in the range of 12.5g/dl -18.5g/dl and females with hemoglobin in the range of 12.5g/dl -16.5g/dl were considered fit for donation. Results: We analyzed that haemoglobin levels measured by HemoCue Hb 201+ were higher than those measured by automated haematologyanalyser. We found a significant correlation between the standard method and HemoCue Hb 201+ for haemoglobin levels less than 12.5 g/dL. However the correlation was found to be less for high haemoglobin levels &gt;18.5g/dl. Sixty percent of donors, who had haemoglobin levels higher than upper limit were unnecessarily rejected accounting for high deferral rate. Conclusion: We concluded that HemoCue Hb 201+ was suitable for screening for low haemoglobin in blood donors. Male predominance among blood donors and topography of this particular area accounts for donors with haemoglobin values near upper limit which were falsely deferred on the basis of HemoCue Hb 201+. Also testing with this method was found to be expensive, so it was recommended that qualitative methods should be used for primary screening among blood donors and accurate quantitative methods to be used only when qualitative methods fail.

Preclinical Drug Response Metric Based on Cellular Response Phenotype Provides Better Pharmacogenomic Variables with Phenotype Relevance.

High-throughput screening of drug response in cultured cell lines is essential for studying therapeutic mechanisms and identifying molecular variants associated with sensitivity to drugs. Assessment of drug response is typically performed by constructing a dose-response curve of viability and summarizing it to a representative, such as IC50. However, this is limited by its dependency on the assay duration and lack of reflections regarding actual cellular response phenotypes. To address these limitations, we consider how each response-phenotype contributes to the overall growth behavior and propose an alternative method of drug response screening that takes into account the cellular response phenotype. In conventional drug response screening methods, the ranking of sensitivity depends on either the metric used to construct the dose-response curve or the representative factor used to summarize the curve. This ambiguity in conventional assessment methods is due to the fact that assessment methods are not consistent with the underlying principles of population dynamics. Instead, the suggested phenotype metrics provide all phenotypic rates of change that shape overall growth behavior at a given dose and better response classification, including the phenotypic mechanism of overall growth inhibition. This alternative high-throughput drug-response screening would improve preclinical pharmacogenomic analysis and the understanding of a therapeutic mechanism of action.

PP148 Liquid Biopsy For The Detection Of Ovarian Or Endometrial Cancer In Samples Taken From The Pap Smear: PapSEEK

IntroductionThe PapSEEK test is an emerging minimally invasive technique in which samples are collected from the endocervical or intrauterine cavity with the Papanicolaou (Pap) brush or the Tao brush to detect somatic mutations or aneuploidies indicating the presence of endometrial or ovarian cancer.MethodsWe systematically searched for articles published up to October 2020 in the following electronic databases: Medline, Embase, the Cochrane Library, and the Centre for Reviews and Dissemination. We included experimental studies, observational primary studies, and cost-effectiveness studies evaluating the safety, effectiveness, and cost effectiveness of the PapSEEK test for the early detection of ovarian or endometrial cancer. Relevant outcomes included sensitivity, specificity, the coefficient of variation, re-test rates, the incremental cost-effectiveness ratio, the incremental cost-utility ratio, and the cost of each alternative.ResultsA single relevant retrospective study was identified. In this study, samples from women with endometrial cancer (n = 656) and ovarian cancer (n = 254) were collected with the Pap brush and Tao brush and compared with samples from healthy women (n = 1,002). The diagnostic validity for somatic mutation or aneuploidies obtained with a Pap brush had a sensitivity of 81% for endometrial cancer and 33% for ovarian cancer, and a specificity of 99% for both conditions. When samples were collected from the intrauterine cavity with a Tao brush, the sensitivity increased to 93% for endometrial cancer and to 45% for ovarian cancer. The sensitivity of the PapSEEK test increased only for ovarian cancer when plasma samples to detect circulating tumor DNA were collected in addition to Pap smear samples. This strategy provided a diagnostic validity of 43%, which was higher in late-stage ovarian cancer (56% versus 35%), and a specificity of 100%.ConclusionsProspectively designed studies are required to assess the safety and effectiveness of the PapSEEK test in screening settings, as well as studies comparing the technology with conventional screening methods. No cost-effectiveness studies have been conducted for the PapSEEK test.

Improved phenotyping procedure for evaluating resistance in rice against gall midge (Orseolia oryzae, Wood-Mason)

BackgroundThe rice gall midge (RGM, Orseolia oryzae, Wood-Mason), an important stem-feeding pest worldwide, has caused serious production losses over the past decades. Rice production practices indicate that the most reliable method for managing RGM is the deployment of cultivars that incorporate host resistance. However, the conventional phenotypic screening method of rice resistance to RGM suggested by the International Rice Research Institute (IRRI) has been used for approximately 30 years, and only 12 rice varieties/lines (including controls) can be evaluated in one tray. It is not suitable for high-throughput phenotyping of rice germplasm. Moreover, a suitable method to prepare samples for molecular biological studies of rice resistance against RGM is imperative with the rapid development of modern molecular techniques.ResultsThe proper density of seedlings/RGM was determined for four seeding arrangements. A high-throughput phenotyping method (HTPM) for 60 lines/varieties infested with 36 female RGM adults in one tray, as described by method 4–3 (seeded 60 lines/varieties), was developed and verified using mutant screening. Furthermore, one RGM resistance gene flanked by markers 12RM28346 and 12RM28739 on chromosome 12 was simultaneously detected using method 2–2 (seeded 30 lines/varieties in one tray) treated with 24 RGM and analyzed using conventional and simplified grading systems. Genetic analysis of the RGM resistance gene was confirmed using a method identical to that suggested by IRRI. Finally, one bucket with 24 seedlings treated with at least five female RGM adults was efficacious and could offer adequate samples for insect development observation or molecular biological studies.ConclusionA highly efficient and reliable procedure for evaluation of resistance in rice to RGM was developed and improved, and was verified through mutant screening, gene mapping, genetic analysis, and insect growth and development observations.

Comparison of Time Taken to Assess Cognitive Function Using a Fully Immersive and Automated Virtual Reality System vs. the Montreal Cognitive Assessment.

Introduction: Dementia is increasingly prevalent globally. Existing questionnaire-based cognitive assessment tools may not comprehensively assess cognitive function and real-time task-performance across all cognitive domains. CAVIRE (Cognitive Assessment by VIrtual REality), a fully immersive virtual reality system incorporating automated audio-visual instructions and a scoring matrix was developed to assess the six cognitive domains, with potential to maintain consistency in execution of the testing environment and possibly time-saving in busy primary care practice.Aims: This is a feasibility study to compare the completion times of the questionnaire-based Montreal Cognitive Assessment (MoCA) and the CAVIRE in cognitively-healthy Asian adults aged between 35 and 74 years, overall, and in and across each 10-year age group (35–44; 45–54; 55–64; 65–74).Methods: A total of 100 participants with a MoCA score of 26 or more were recruited equally into the four 10-year age groups at a primary care clinic in Singapore. Completion time for the MoCA assessment for each participant was recorded. They were assessed using the CAVIRE, comprising 13 segments featuring common everyday activities assessing all six cognitive domains, and the completion time was also recorded through the embedded automated scoring and timing framework.Results: Completion time for CAVIRE as compared to MoCA was significantly (p < 0.01) shorter, overall (mean difference: 74.9 (SD) seconds) and in each age group. Younger, vs. older, participants completed both the MoCA and CAVIRE tasks in a shorter time. There was a greater variability in the completion time for the MoCA, most markedly in the oldest group, whereas completion time was less variable for the CAVIRE tasks in all age groups, with most consistency in the 45–54 year-age group.Conclusion: We demonstrate almost equivalent completion times for a VR and a questionnaire-based cognition assessment, with inter-age group variation in VR completion time synonymous to that in conventional screening methods. The CAVIRE has the potential to be an alternative screening modality for cognition in the primary care setting.

A Metagenomic Nanopore Sequence Analysis Combined with Conventional Screening and Spectroscopic Methods for Deciphering the Antimicrobial Metabolites Produced by Alcaligenes faecalis Soil Isolate MZ921504.

The continuous development of multidrug resistance pathogens with limited therapeutic options has become a great problem globally that impose sever health hazards. Accordingly, searching for of new antimicrobials became an urgent demand and great challenge. Soil significantly have been associated with several species that are antibiotic producers. In this study, combination of conventional screening methods with Liquid chromatography- Mass spectroscopy (LC/MS) and metagenomic nanopore sequence analysis have been conducted for the deciphering the active metabolites produced by soil isolate(s). Preliminary soil screening resulted in a Gram-negative isolate identified via 16S ribosomal RNA as Alcaligenes faecalis isolate MZ921504 with promising antimicrobial activities against wide range of MDR gram-positive and gram-negative pathogens. The LC/MS analysis of the metabolites of A. faecalis isolate MZ921504 confirmed the presence of ectoine, bacillibactin, quinolobactin and burkholderic acid. Metagenomics sequence analysis of the soil sample (NCBI GenBank accession PRJNA771993) revealed the presence of conserved biosynthetic gene clusters of ectoine, bacteriocin, bacillibactin, quinolobactin, terpene and burkholderic acid of A. faecalis. In conclusion, A. faecalis isolate MZ921504 is a promising source for antimicrobial metabolites. LC/MS spectral analysis and third generation sequencing tools followed by secondary metabolite gene clusters analysis are useful methods to predict the nature of the antimicrobial metabolites.

Anti-Cancer Drug Screening with Microfluidic Technology

The up-and-coming microfluidic technology is the most promising platform for designing anti-cancer drugs and new point-of-care diagnostics. Compared to conventional drug screening methods based on Petri dishes and animal studies, drug delivery in microfluidic systems has many advantages. For instance, these platforms offer high-throughput drug screening, require a small number of samples, provide an in vivo-like microenvironment for cells, and eliminate ethical issues associated with animal studies. Multiple cell cultures in microfluidic chips could better mimic the 3D tumor environment using low reagents consumption. The clinical experiments have shown that combinatorial drug treatments have a better therapeutic effect than monodrug therapy. Many attempts have been made in this field in the last decade. This review highlights the applications of microfluidic chips in anti-cancer drug screening and systematically categorizes these systems as a function of sample size and combination of drug screening. Finally, it provides a perspective on the future of the clinical applications of microfluidic systems for anti-cancer drug development.