Biological Modification Research Articles

StratosPHere 2: study protocol for a response-adaptive randomised placebo-controlled phase II trial to evaluate hydroxychloroquine and phenylbutyrate in pulmonary arterial hypertension caused by mutations in BMPR2

BackgroundPulmonary arterial hypertension is a life-threatening progressive disorder characterised by high blood pressure (hypertension) in the arteries of the lungs (pulmonary artery). Although treatable, there is no known cure for this rare disorder, and its exact cause is unknown. Mutations in the bone morphogenetic protein receptor type-2 (BMPR2) are the most common genetic cause of familial pulmonary arterial hypertension. This study represents the first-ever trial of treatments aimed at directly rescuing the BMPR2 pathway, repurposing two drugs that have shown promise at restoring levels of BMPR2 signalling: hydroxychloroquine and phenylbutyrate.MethodsThis three-armed phase II precision medicine study will investigate BMPR2 target engagement and explore the efficacy of two repurposed therapies in pulmonary arterial hypertension patients with BMPR2 mutations. Patients will be stratified based on two BMPR2 mutation classes: missense and haploinsufficient mutations. Eligible subjects will be randomised to one of the three arms (two active therapy arms and a placebo arm, all plus standard of care) following a Bayesian response-adaptive design implemented independently in each stratum and updated in response to a novel panel of primary biomarkers designed to assess biological modification of the disease.DiscussionThe results of this trial will provide the first randomised evidence of the efficacy of these therapies to rescue BMPR2 function and will efficiently explore the potential for a differential response of these therapies per mutation class to address causes rather than consequences of this rare disease.Trial registrationThe study has been registered with ISRCTN (ISRCTN10304915, 22/09/2023).

Escherichia coli on colorectal cancer: A two-edged sword.

Escherichia coli (E. coli) is a ubiquitous symbiotic bacterium in the gut, and the diversity of E. coli genes determines the diversity of its functions. In this review, the two-edged sword theory was innovatively proposed. For the question 'how can we harness the ambivalent nature of E. coli to screen and treat CRC?', in terms of CRC screening, the variations in the abundance and subtypes of E. coli across different populations present an opportunity to utilise it as a biomarker, while in terms of CRC treatment, the natural beneficial effect of E. coli on CRC may be limited, and engineered E. coli, particularly certain subtypes with probiotic potential, can indeed play a significant role in CRC treatment. It seems that the favourable role of E. coli as a genetic tool lies not in its direct impact on CRC but its potential as a research platform that can be integrated with various technologies such as nanoparticles, imaging methods, and synthetic biology modification. The relationship between gut microflora and CRC remains unclear due to the complex diversity and interaction of gut microflora. Therefore, the application of E. coli should be based on the 'One Health' view and take the interactions between E. coli and other microorganisms, host, and environmental factors, as well as its own changes into account. In this paper, the two-edged sword role of E. coli in CRC is emphasised to realise the great potential of E. coli in CRC screening and treatment.

Integrative approaches to improve litchi (Litchi chinensis Sonn.) plant health using bio-transformations and entomopathogenic fungi

Bio-transformations refer to the chemical modifications made by an organism on a chemical compound that often involves the interaction of plants with microbes to alter the chemical composition of soil or plant. Integrating bio-transformations and entomopathogenic fungi into litchi cultivation can enhance symbiotic relationships, microbial enzymatic activity in rhizosphere, disease suppression and promote overall plant health. The integration of biological formulations and entomopathogenic fungi can significantly influence growth, nutrient dynamics, physiology, and rhizosphere microbiome of air-layered litchi (Litchi chinensis Sonn.) saplings. Biological modifications included, K-mobilizers, AM fungi, Pseudomonas florescence and Azotobacter chroococcum along with Metarhizium, entomopathogenic fungi have been used. The treatments included, T1-Litchi orchard soil + sand (1:1); T2-Sand + AM fungi + Azotobacter chroococcum (1:2:1); T3-Sand + Pseudomonas florecence + K-mobilizer (1:1:1); T4- AM fungi + K-mobilizers (1:1); T5, P. Florecence + A. chroococcum + K-mobilizer (1:1:1); T6-Sand + P. florecence (1:2) and T7-Uninoculated control for field performance. Treatments T4-T6 were further uniformly amended with drenching of Metarrhizium in rhizosphere. T2 application significantly increased resident microbe survival, total chlorophyll content and root soil ratio in seedlings. A. chroococcum, Pseudomonas, K-mobilizers and AM fungi increased in microbial biomass of 2.59, 3.39, 2.42 and 2.77 times, respectively. Acidic phosphatases, dehydrogenases and alkaline phosphatases were increased in rhizosphere. Leaf nutrients reflected through DOP were considerably altered by T2 treatment. Based on Eigen value, PCA-induced changes at biological modifications showed maximum total variance. The study inferred that the bio-transformations through microbial inoculants and entomopathogenic fungi could be an encouraging strategy to enhance the growth of plants, health and productivity. Such practices align well with the goals of sustainable agriculture through biological means by reducing dependency on chemical inputs. By delving into these aspects, the research gaps including microbial processes, competitive and symbiotic relationships, resistance in microbes and how complex interactions among bio-transformations, entomopathogenic fungi and microbes can significantly impact the health and productivity of litchi. Understanding and harnessing these interactions can lead to more effective and sustainable farming practices.

Heterologous expression of bacterial tyrosinase and its applications in biological hair dyeing and DOPA modification of hydrolyzed silk fibroin

Tyrosinase is a copper-containing polyphenol oxidase widely applied in the food, cosmetics, pharmaceutical, and other industries. Currently, the production of commercial tyrosinase primarily relies on extraction from fungi, which has high costs, low purity, low specific activity, and poor stability. The objective of this study is to obtain highly expressed bacterial tyrosinase with potential for industrial applications. The bacterial tyrosinases from five different sources were heterologously expressed in Escherichia coli BL21(DE3), and the tyrosinases TyrBm and TyrVs derived from Bacillus megaterium and Verrucomicrobium spinosum were obtained with the enzyme activities of (16.1±0.2) U/mL and (48.6±0.9) U/mL, respectively. After protein purification, we compared the enzymatic properties of TyrBm and TyrVs, which revealed that TyrVs exhibited better thermal stability and higher substrate specificity than TyrBm. On the basis of characterizing TyrVs with high catalytic performance, we established a biological hair dyeing system based on TyrVs catalysis to achieve in-situ catalytic hair dyeing. The color washing fastness test measured the ∆E value less than 7.38±0.64 after simulated 14-day cleaning. To facilitate the rapid separation of catalytic products and enzymes, we successfully constructed an immobilized enzyme TyrVs-CipA dependent on self-assembly label CipA and applied this enzyme in the DOPA modification of hydrolyzed silk fibroin (HSF). The immobilized enzyme continuously catalyzed HSF for more than seven cycles, resulting in a single DOPA modification degree exceeding 70.00%. Further investigations demonstrated that DOPA modification enhances the scavenging activity of HSF towards DPPH and O2- radicals by 507.80% and 78.23%, respectively. This study provides a technical foundation for the development of environmentally friendly biological hair dye based on tyrosinase and biomaterials for tissue engineering.

In-depth Investigation into Supervised and Unsupervised Models within Artificial Neural Networks

An Artificial Neural Network (ANN) is an information-handling worldview that is jazzed up by exactly how natural nerves, like the mind, procedure details. The essential element of this particular worldview is the initial Building of the information dealing with the unit. A neural network is a difficult development that comprises a gathering of interconnected nerve cells, which offers a remarkably electrifying possibility unlike complicated essential reasoning and also various other treatments which may think a notable component in today's software engineering industry, thus professionals from different fields are organizing the synthetic neural networks to tackle the concerns of trend awareness, prophecy, optimization, associative moment and command. It is constructed of an enormous lot of specially connected taking care of elements (neurons) functioning as one to address explicit issues. ANNs, identical to individuals, innovation as an aesthetic presentation. An ANN is developed for a specific app, for example, pattern recognition or records classification, using developing knowledge. Discovering in biological units features modifications following the synaptic associations between neurons.

Progress in the Development of Antifouling Electrochemical Biosensors

Electrochemical biosensors a subclass of biosensors, consisting of a biosensing element and an electrochemical transducer, have been widely used in various fields due to their excellent performance and portable device. However, in complex actual samples, non-specific adsorption of proteins and solid particles, and adhesion of cells and bacteria will lead to problems such as reduced sensor sensitivity, prolonged response time, and expanded detection errors. Therefore, constructing antifouling sensing platforms to effectively resist the bioadhesion of non-targets is crucial for the performance of biosensors. This study first introduces the commonly used classifications of electrochemical biosensors and their main contaminants. It also provides a comprehensive overview of the construction methods and application research of electrochemical antifouling sensors using different strategies, including the construction of physical, chemical and biological modification interfaces. In addition, the research progress on antifouling and antibacterial dual-action coatings for electrochemical detection is also reviewed. Finally, the challenges and future development trends of various methods are summarized, providing clues for better practical applications of electrochemical biosensors.

Emulsifying properties of egg proteins: Influencing factors, modification techniques, and applications.

As an essential food ingredient with good nutritional and functional properties and health benefits, eggs are widely utilized in food formulations. In particular, egg proteins have good emulsification properties and can be commonly used in various food products, such as mayonnaise and baked goods. Egg protein particles can act as stabilizers for Pickering emulsions because they can effectively adsorb at the oil-water interface, reduce interfacial tension, and form a stable physical barrier. Due to their emulsifying properties, biocompatibility, controlled release capabilities, and ability to protect bioactive substances, egg proteins have become ideal carriers for encapsulating and delivering functional substances. The focus of this review is to summarize current advances in using egg proteins as emulsifiers. The effects of influencing factors (temperature, pH, and ionic strength) and various modification methods (physical, chemical, and biological modification) on the emulsifying properties of egg proteins are discussed. In addition, the application of egg proteins as emulsifiers in food products is presented. Through in-depth research on the emulsifying properties of egg proteins, the optimization of their applications in food, biomedical, and other fields can be achieved.

A novel m6A reader RBFOX2 expression is increased in oral squamous cell carcinoma and promotes tumorigenesis

Background and ObjectiveOral squamous cell carcinoma (OSCC) is a significant global health concern due to its aggressive nature and poor prognosis. Recent research has highlighted the important role of RNA modifications in cancer biology, especially N6-methyladenosine (m6A) modifications, which are controlled by a complex interplay of m6A regulators. This study specifically investigates RBFOX2, a new m6A reader, and its involvement in OSCC. MethodsOur study primarily utilized OSCC tissue, adjacent normal tissue samples, OSCC cell lines, and normal healthy oral keratinocytes to validate RBFOX2 mRNA expression. Additionally, we used the TCGA-HNSCC dataset for large cohort analysis and clinicopathological characterization. Furthermore, we visualized the RBFOX2 network to identify the primary functions of the protein. ResultsOur research shows a noticeable increase in RBFOX2 expression in OSCC tissues compared to adjacent non-tumorous tissues, as determined by quantitative PCR and immunohistochemistry analyses. Functional pathway enrichment analysis revealed that RBFOX2 is involved in the receptor tyrosine kinase signaling pathway and the Hippo signaling pathway, which plays a critical role in oral cancer development and progression. Clinically, elevated RBFOX2 expression correlated with advanced tumor stages and poorer patient outcomes, demonstrating its prognostic value. These findings indicate that RBFOX2 acts as an oncogenic driver in OSCC as an m6A reader, facilitating the expression of m6A-modified oncogenes. ConclusionOur study identifies RBFOX2 as a critical player in OSCC pathogenesis and opens avenues for novel therapeutic strategies targeting the m6A regulatory machinery in this malignancy.

CRISPR/Cas9-based genome engineering in the filamentous fungus Rhizopus oryzae and its application to L-lactic acid production.

The filamentous fungus Rhizopus oryzae is one of the main industrial strains for the production of a series of important chemicals such as ethanol, lactic acid, and fumaric acid. However, the lack of efficient gene editing tools suitable for R. oryzae makes it difficult to apply technical methods such as metabolic engineering regulation and synthetic biology modification. A CRISPR-Cas9 system suitable for efficient genome editing in R. oryzae was developed. Firstly, four endogenous U6 promoters of R. oryzae were identified and screened with the highest transcriptional activity for application to sgRNA transcription. It was then determined that the U6 promoter mediated CRISPR/Cas9 system has the ability to efficiently edit the genome of R. oryzae through NHEJ and HDR-mediated events. Furthermore, the newly constructed CRISPR-Cas9 dual sgRNAs system can simultaneously disrupt or insert different fragments of the R. oryzae genome. Finally, this CRISPR-Cas9 system was applied to the genome editing of R. oryzae by knocking out pyruvate carboxylase gene (PYC) and pyruvate decarboxylase gene (pdcA) and knocking in phosphofructokinase (pfkB) from Escherichia coli and L-lactate dehydrogenase (L-LDH) from Heyndrickxia coagulans, which resulted in a substantial increase in L-LA production. In summary, this study showed that the CRISPR/Cas9-based genome editing tool is efficient for manipulating genes in R. oryzae.

Research on electron beam irradiation in the multiscale structure of starch and its related applications: A review.

Electron beam irradiation (EBI), as a typical "green" emerging technology, can effectively alter the functional properties of starch by influencing its microstructure. This alteration enables starch to meet the current demands of consumers and the market for "health food." This paper reviews studies on modifying various starches using EBI and describes the changes in microstructure, physicochemical properties, and functional properties induced by this method. Additionally, the effects of EBI on starch-containing food products are discussed, along with issues to be addressed and research gaps in the synergistic treatment of modified starch. It is noted that the source, irradiation dose, and irradiation time all influence the effectiveness of starch modification. Given the characteristics of EBI technology, integrating physical, chemical, and biological modification methods can optimize the modification process and enhance efficiency. This technology can potentially diversify modified starch varieties and expand their applications. Furthermore, there remains significant research potential in producing modified starch using EBI technology and applying it to the food industry.

What happens when the insecticide does not kill? A review of sublethal toxicology and insecticide resistance in triatomines.

Chagas disease is considered one of the most important human parasitosis in the United States. This disease is mainly transmitted by insects of the subfamily Triatominae. The chemical vector control is the main tool for reducing the incidence of the disease. However, the presence of triatomines after pyrethroids spraying has been reported in some regions, as in the case of Triatoma infestans in Argentina and Bolivia. The presence of insects can be explained by the colonization from neighbouring areas, the reduction of insecticide dose to sublethal levels due to environmental factors, and/or by the evolution of insecticide resistance. In the last two scenarios, a proportion of the insects is not killed by insecticide and gives rise to residual populations. This article focuses on the toxicological processes associated with these scenarios in triatomines. Sublethal doses may have different effects on insect biology, that is, sublethal effects, which may contribute to the control. In addition, for insect disease vectors, sublethal doses could have negative effects on disease transmission. The study of sublethal effects in triatomines has focused primarily on the sequence of symptoms associated with nervous intoxication. However, the effects of sublethal doses on excretion, reproduction and morphology have also been studied. Rhodnius prolixus and T. infestans and pyrethroids insecticides were the triatomine species and insecticides, respectively, mainly studied. Insecticide resistance is an evolutionary phenomenon in which the insecticide acts as a selective force, concentrating on the insect population's pre-existing traits that confer resistance. This leads to a reduction in the susceptibility to the insecticide, which was previously effective in controlling this species. The evolution of resistance in triatomines received little attention before the 2000s, but after the detection of the first focus of resistance associated with chemical control failures in T. infestans from Argentina in 2002, the study of resistance increased remarkably. A significant number of works have studied the geographical distribution, the resistance mechanisms, the biological modifications associated with resistance, the environmental influences and the genetic of T. infestans resistant to pyrethroid insecticides. Currently, studies of insecticide resistance are gradually being extended to other areas and other species. The aim of this article was to review the knowledge on both phenomena (sublethal effects and insecticide resistance) in triatomines. For a better understanding of this article, some concepts and processes related to insect-insecticide interactions, individual and population toxicology and evolutionary biology are briefly reviewed. Finally, possible future lines of research in triatomine toxicology are discussed.

An Integrated Photonic Biosensing Platform for Pathogen Detection in Aquaculture.

Aquaculture is expected to play a vital role in solving the challenge of sustainably providing the growing world population with healthy and nutritious food. Pathogen outbreaks are a major risk for the sector, so early detection and a timely response are crucial. This can be enabled by monitoring the pathogen levels in aquaculture facilities. This paper describes a photonic biosensing platform based on silicon nitride waveguide technology with integrated active components, which could be used for such applications. Compared to the state of the art, the current system presents improvements in terms of miniaturization of the Photonic Integrated Circuit (PIC) and the development of wafer-level processes for hybrid integration of active components and for material-selective chemical and biological surface modification. Furthermore, scalable processes for integrating the PIC in a microfluidic cartridge were developed, as well as a prototype desktop readout instrument. Three bacterial aquaculture pathogens (Aeromonas salmonicida, Vagococcus salmoninarum, and Yersinia ruckeri) were selected for assay development. DNA biomarkers were identified, corresponding primer-probe sets designed, and qPCR assays developed. The biomarker for Aeromonas was also detected using the hybrid PIC platform. This is the first successful demonstration of biosensing on the hybrid PIC platform.

Epitranscriptomic m5C methylation of SARS-CoV-2 RNA regulates viral replication and the virulence of progeny viruses in the new infection.

While the significance of N6-methyladenosine (m6A) in viral regulation has been extensively studied, the functions of 5-methylcytosine (m5C) modification in viral biology remain largely unexplored. In this study, we demonstrate that m5C is more abundant than m6A in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and provide a comprehensive profile of the m5C landscape of SARS-CoV-2 RNA. Knockout of NSUN2 reduces m5C levels in SARS-CoV-2 virion RNA and enhances viral replication. Nsun2 deficiency mice exhibited higher viral burden and more severe lung tissue damages. Combined RNA-Bis-seq and m5C-MeRIP-seq identified the NSUN2-dependent m5C-methylated cytosines across the positive-sense genomic RNA of SARS-CoV-2, and the mutations of these cytosines enhance RNA stability. The progeny SARS-CoV-2 virions from Nsun2 deficiency mice with low levels of m5C modification exhibited a stronger replication ability. Overall, our findings uncover the vital role played by NSUN2-mediated m5C modification during SARS-CoV-2 replication and propose a host antiviral strategy via epitranscriptomic addition of m5C methylation to SARS-CoV-2 RNA.

DeepPGD: A Deep Learning Model for DNA Methylation Prediction Using Temporal Convolution, BiLSTM, and Attention Mechanism.

As part of the field of DNA methylation identification, this study tackles the challenge of enhancing recognition performance by introducing a specialized deep learning framework called DeepPGD. DNA methylation, a crucial biological modification, plays a vital role in gene expression analyses, cellular differentiation, and the study of disease progression. However, accurately and efficiently identifying DNA methylation sites remains a pivotal concern in the field of bioinformatics. The issue addressed in this paper is the presence of methylation in DNA, which is a binary classification problem. To address this, our research aimed to develop a deep learning algorithm capable of more precisely identifying these sites. The DeepPGD framework combined a dual residual structure involving Temporal convolutional networks (TCNs) and bidirectional long short-term memory (BiLSTM) networks to effectively extract intricate DNA structural and sequence features. Additionally, to meet the practical requirements of DNA methylation identification, extensive experiments were conducted across a variety of biological species. The experimental results highlighted DeepPGD's exceptional performance across multiple evaluation metrics, including accuracy, Matthews' correlation coefficient (MCC), and the area under the curve (AUC). In comparison to other algorithms in the same domain, DeepPGD demonstrated superior classification and predictive capabilities across various biological species datasets. This significant advancement in algorithmic prowess not only offers substantial technical support, but also holds potential for research and practical implementation within the DNA methylation identification domain. Moreover, the DeepPGD framework shows potential for application in genomics research, biomedicine, and disease diagnostics, among other fields.

Current trends in additive manufacturing of selective laser melting for biomedical implant applications

Additive manufacturing (AM) has obtained great attentions in the productions of customizable implants with complex shapes and good mechanical properties. AM fabricated implants of Ti alloy (Ti), stainless steel (SS), cobalt chromium (CoCr), iron (Fe) have been revolutionized in the productions of medical instruments, such as in artificial organs, including heart stents, drug delivery, scaffolds and orthopedic implants. However, high Young's modulus, superior mechanical characteristic, lack of surface integration, high corrosions, failed bone remodeling, and antibacterial performance are still incomparable to be used in anatomical structure. This review studied the issue of metallic implants designed by powder bed fusion of selective laser melting (SLM) techniques on the surface morphology, stress shielding effects, osseointegration, and antibacterial capabilities owing to implant failure. The rate of degradation and corrosion resistance of the metallic SLM implant, which would have an impact on cell proliferation and bone remodeling between the surrounding tissue, were discussed. The current review article also highlighted the various SLM parameters, like mechanical, chemical, and biological modifications, that might enhance the implant. Finally, applications, challenges, opportunities, and future developments of metallic implants using SLM techniques were addressed.

Bacterial imaging in tumour diagnosis.

Some bacteria, such as Escherichia coli (E. coli) and Salmonella typhimurium (S. typhimurium), have an inherent ability to locate solid tumours, making them a versatile platform that can be combined with other tools to improve the tumour diagnosis and treatment. In anti-cancer therapy, bacteria function by carrying drugs directly or expressing exogenous therapeutic genes. The application of bacterial imaging in tumour diagnosis, a novel and promising research area, can indeed provide dynamic and real-time monitoring in both pre-treatment assessment and post-treatment detection. Different imaging techniques, including optical technology, acoustic imaging, magnetic resonance imaging (MRI) and nuclear medicine imaging, allow us to observe and track tumour-associated bacteria. Optical imaging, including bioluminescence and fluorescence, provides high-sensitivity and high-resolution imaging. Acoustic imaging is a real-time and non-invasive imaging technique with good penetration depth and spatial resolution. MRI provides high spatial resolution and radiation-free imaging. Nuclear medicine imaging, including positron emission tomography (PET) and single photon emission computed tomography (SPECT) can provide information on the distribution and dynamics of bacterial population. Moreover, strategies of synthetic biology modification and nanomaterial engineering modification can improve the viability and localization ability of bacteria while maintaining their autonomy and vitality, thus aiding the visualization of gut bacteria. However, there are some challenges, such as the relatively low bacterial abundance and heterogeneously distribution within the tumour, the high dimensionality of spatial datasets and the limitations of imaging labeling tools. In summary, with the continuous development of imaging technology and nanotechnology, it is expected to further make in-depth study on tumour-associated bacteria and develop new bacterial imaging methods for tumour diagnosis.

Exposure-response relationship of guselkumab and the potential of serum proteomics in identifying predictive biomarker candidates in psoriasis.

Response to biologics in psoriasis varies in real-world settings. Serum biomarkers could aid biologic selection and dose modifications to improve patient outcomes while encouraging cost-effective care. To explore the exposure-response relationship for guselkumab (GUS), to define a GUS concentration target for optimal response and to evaluate the potential of serum protein levels as predictive biomarker candidates. This is a prospective, multicentric, cohort study in psoriasis patients treated with GUS. Serum GUS trough concentrations (TCs) collected at multiple timepoints were measured using an in-house immunoassay. Next, proximity extension assay technology (Target 96 Inflammation Panel Olink®) was used to measure serum protein levels in a subcohort including 38 GUS patients (week 0 and week 4), six psoriasis patients naive for systemic treatment and four healthy controls. Seventy-five patients participated and 400 samples were collected. Guselkumab TCs and clinical response were correlated at week 4, week 12 and in steady-state (≥20 weeks). Optimal responders (Psoriasis Area and Severity Index [PASI] ≤ 2) had significantly higher TCs than suboptimal responders from week 4 onwards in treatment. An optimal steady-state TC of 1.6 μg/mL was defined. Although TC and absolute PASI were lower and worse, respectively, in patients weighing ≥90 kg, clinical outcomes referred to desirable to excellent PASI ranges. Therefore, we do not recommend systematically higher GUS doses in obese patients. We could not reveal early differentially expressed proteins to distinguish future optimal from suboptimal responders. We demonstrated an exposure-response relationship for GUS and an optimal steady-state TC of 1.6 μg/mL in real-world psoriasis patients. Hereby, we deliver more evidence that therapeutic drug monitoring poses a promising strategy in optimizing GUS treatment. No biomarker candidates were identified through serum proteomics. We propose protein screening should be repeated in larger cohorts to continue the quest for predictive biomarkers.

Unraveling the influence of nitration on pore formation time in electroporation of cell membranes: a molecular dynamics simulation approach

Electroporation, the transient permeabilization of cell membranes induced by electric fields, is an essential technique in biomedicine, facilitating gene delivery, drug transport, and cancer therapy. Despite its wide application, the influence of nitration, a biological modification involving the addition of nitro groups to phospholipids, on electroporation dynamics remains understudied. Here, we employ molecular dynamics simulations to investigate the impact of nitration on pore formation during electroporation. By systematically varying nitration levels and electric field strengths, we explore the nuanced interplay between nitration and electroporation kinetics. Our simulations reveal that increasing nitration levels significantly accelerate pore formation, with notable reductions in pore formation times observed at higher nitration percentages and stronger electric fields. This phenomenon underscores the modulatory role of nitration in altering the dynamics of electroporation. Additionally, our study sheds light on the intricate mechanisms underlying this process, providing essential insights for optimizing electroporation protocols in gene therapy, drug delivery, plasma cancer treatment and related biomedical applications. These findings illuminate the synergistic relationship between nitration and electroporation, paving the way for future advancements in this vital field.

Label/immobilization-free Cas12a-based electrochemiluminescence biosensor for sensitive DNA detection

Electrochemiluminescence (ECL) is one of the most sensitive techniques in the field of diagnostics. However, they typically require luminescent labeling and electrode surface biological modification, which is a time-consuming and laborious process involving multiple steps and may also lead to low reaction efficiency. Fabricating label/modification-free biosensors has become one of the most attractive parts for simplifying the ECL assays. In this work, the ECL luminophores carbon dots (CDs) were encapsulated in DNA hydrogel in situ by a simple rolling circle amplification (RCA) reaction. Upon binding of the target DNA, active Cas12a induces a collateral cleavage of the hydrogel's ssDNA backbone, resulting in a programmable degradation of the hydrogel and the release of CDs. By directly measuring the released CDs ECL, a simple and rapid label/modification-free detection of the target HPV-16 was realized. It is noted that this method allowed for 0.63 pM HPV-16 DNA detection without any amplification step, and it could take only ∼60 min for a fast test of a human serum sample. These results showed that our label/modification-free ECL biosensor has great potential for use in simple, rapid, and sensitive point-of-care (POC) detection.

The biological modifications of milk are linked to mental health of mothers of infants affected by bronchiolitis

IntroductionBreast milk is a dynamic type of nourishment that changes based on the needs of the child. An increasing amount of data suggests that mental health may be an important factor in such modulation. In addition, breast milk contains extracellular vesicles (EVs), which are currently considered an important dynamic system of communication between cells, even of different individuals.ObjectivesPurpose of this article is to investigate whether changes in breast milk in terms of EVs concentrations are related to maternal mental health.MethodsThis is a case-control study for which we enrolled mothers of infants with bronchiolitis (N=33) and mothers of healthy infants (N=13). Breast milk samples were taken and EVs concentrations were quantified. Maternal mental health was assessed by administration of five different psychometric scales: Edinburgh Postnatal Depression Scale (EPDS), State Trait Anxiety Inventory (STAI-S, STAI-T), Barkin Index of Maternal Functioning (BIMF), The Connor-Davidson Resilience Scale 10 items (CD-RISC). Subsequently, scale scores were related to evs concentrations by negative binomial regressions adjusted for case-control.ResultsAs maternal resilience increases, the EVs of neutrophilic origin (p=0.0447) and those of endothelial origin (p=0.0078) decrease¹. In contrast, an increased EPDS score is associated with higher levels of B-lymphocyte EVs (p=0.0376). Scores on the STAI-S scale impact many more populations of EVs²: we observed an increased Incidence Rate Ratio (IRR) of neutrophil-derived EVs (p<0.0001), T-lymphocyte- derived EVs (p=0.0214), NK-cell-derived EVs (p=0.0202), T-reg CD4+ CD25+ (p=0.0141) and endothelial marked EVs (p=0.0180). An increase in STAI-T scale scores also was associated with a significant increase in CD177+ neutrophil-derived EVs (p=0.0028) and endothelial-derived EVs (p=0.0111)³.Image:Image 2:Image 3:ConclusionsEVs concentrations in breast milk are associated with maternal mental health. Specifically, stress and related severity of anxiety is able to increase the concentrations of EVs derived from inflammatory cells, which suggests an increase in their number and activity. Further research is needed to confirm these preliminary findings.Disclosure of InterestNone Declared