Engineering of CRISPR-Cas PAM recognition using deep learning of vast evolutionary data.

CRISPR-Cas enzymes must recognize a protospacer-adjacent motif (PAM) to edit a genomic site, significantly limiting the range of targetable sequences in a genome. Machine learning-based protein engineering provides a powerful solution to efficiently generate Cas protein variants tailored to recognize specific PAMs. Here, we present Protein2PAM, an evolution-informed deep learning model trained on a dataset of over 45,000 CRISPR-Cas PAMs.

Bifunctional hierarchical WO nanorods@Br-doped g-CN Z-scheme heterojunctions for efficient tetracycline photodegradation: Optimization of key parameters, toxicological assessment, and electrocatalytic hydrogen evolution reaction.

In this study, graphitic carbon nitride (CN) and tungsten trioxide (WO) were successfully incorporated into bromine (Br)-doped graphitic carbon nitride (BCN) using an in-situ hydrothermal method. The photocatalytic efficiency of the resulting WO/Br-doped CN (WBCN) composites for the removal of tetracycline (TC) antibiotics under sunlight irradiation was evaluated. The mass ratio of WO to Br-doped CN (BCN) significantly influenced TC adsorption and photocatalytic degradation, with an optimal ratio of 9:1.

Pathological In Vivo Analysis of Helicobacter DNA Infection in Stomach Cells Using Carbon Nanotube Microsensor.

The WHO has classified Helicobacter pylori as a group 1 carcinogen for stomach cancer since early 1994. However, despite the high prevalence of Helicobacter pylori infection, only about 3% of infected people eventually develop gastric cancer.Biomolecular detections of Helicobacter pylori(HP) were compared using specially modified sensors and fluorine immobilized on a carbon nanotube (HFCNT) electrode, which yielded sensitive results.

Maximizing the clinical utility and performance of cytology samples for comprehensive genetic profiling.

Comprehensive molecular profiling by next-generation sequencing has revolutionized tumor classification and biomarker evaluation. However, routine implementation is challenged by the scant nature of diagnostic material obtained through minimally invasive procedures. Here, we describe our long-term experience in profiling cytology samples with an in-depth assessment of the performance, quality metrics, biomarker identification capabilities, and potential pitfalls.

N-functionalized calcium alginate encapsulated with microalgae for efficient Pb removal: Mechanisms and performance in batch and column operations.

In this study, a novel adsorbent called Ca@SP was developed by immobilizing microalgae protein (Spirulina platensis, SP) in an alginate matrix for enhanced Pb²⁺ removal from aqueous solutions. Synthesized via in situ crosslinking, Ca@SP leverages the synergistic effects of alginate's gel-forming ability and SP's N-rich biomass. Characterization of Ca@SP revealed a green spherical hydrogel with a BET specific surface area of 159.

Iodide Enhances the Production of Pseurotin D over Pseurotin A by Inverting the Preference for the S2 versus the S2' Product in the Final Nonenzymatic Step.

Nonenzymatic reactions, though critical in natural product biosynthesis, are significantly challenging to control. Adding 3% NaI to the culture medium of significantly increased pseurotin D () production and decreased pseurotin A () production. Previously, and were suggested to be produced via a nonenzymatic reaction, where the epoxide at C-10 undergoes S2 () or S2' () reactions.

Co-immunization with spike and nucleocapsid based DNA vaccines for long-term protective immunity against SARS-CoV-2 Omicron.

The continuing evolution of SARS-CoV-2 variants challenges the durability of existing spike (S)-based COVID-19 vaccines. We hypothesized that vaccines composed of both S and nucleocapsid (N) antigens would increase the durability of protection by strengthening and broadening cellular immunity compared with S-based vaccines. To test this, we examined the immunogenicity and efficacy of wild-type SARS-CoV-2 S- and N-based DNA vaccines administered individually or together to K18-hACE2 mice.

Microengineered in vitro CAR T cell screens and assays.

Established and emergent microengineered in vitro systems enable the evaluation of chimeric antigen receptor (CAR) T cell product purity, avidity, and functionality. Here, we describe such systems and how they have been used to optimize CAR T cell products by selecting highly viable cells, eliminating off-target cells, and tailoring avidity to balance efficacy and safety. The future of CAR T cell therapy development and manufacturing is expected to be anchored in a cyclical process that integrates multiple high-throughput and patient-centered techniques for identifying, enriching, and evaluating T cell subtypes.

Immunogenic recombinant Mayaro virus-like particles present natively assembled glycoprotein.

Virus-like particles (VLPs) are an established vaccine platform and can be strong immunogens capable of eliciting both humoral and cellular immune responses against a range of pathogens. Here, we show by cryo-electron microscopy that VLPs of Mayaro virus, which contain envelope glycoproteins E1-E2 and capsid, exhibit an architecture that closely resembles native virus. In contrast to monomeric and soluble envelope 2 (E2) glycoprotein, both VLPs as well as the adenovirus and modified vaccinia virus Ankara (MVA) vaccine platforms expressing the equivalent envelope glycoproteins E1-E2, and capsid induced highly neutralising antibodies after immunisation.

Efficacy of ETB-F01, Heat-Killed Strain EB-AMDK19, in Patients with Respiratory Symptoms: A Multicenter Clinical Trial.

Respiratory symptoms are prevalent in the general population, and they are associated with a decline in lung function and increased mortality. The gut-lung connection suggests intestinal dysbiosis may impact lung diseases, with showing promise in regulating extraintestinal diseases. However, its application in patients with respiratory symptoms lacks clinical trial evidence.

MIBiG 4.0: advancing biosynthetic gene cluster curation through global collaboration.

Specialized or secondary metabolites are small molecules of biological origin, often showing potent biological activities with applications in agriculture, engineering and medicine. Usually, the biosynthesis of these natural products is governed by sets of co-regulated and physically clustered genes known as biosynthetic gene clusters (BGCs). To share information about BGCs in a standardized and machine-readable way, the Minimum Information about a Biosynthetic Gene cluster (MIBiG) data standard and repository was initiated in 2015.

Polygenic Prediction of Recurrent Events After Early-Onset Myocardial Infarction.

Background: Myocardial infarction (MI) is a complex disease caused by both lifestyle and genetic factors. This study aims to investigate the predictive value of genetic risk, in addition to traditional cardiovascular risk factors, for recurrent events following early-onset MI.

Methods: The Italian Genetic Study of Early-Onset Myocardial Infarction is a cohort study enrolling patients with MI before 45 years.

Long-Term Outcome of Rescue Stenting for Acute Intracranial Atherosclerotic Stenosis Related Large Vessel Occlusion in Anterior Circulation.

Background And Purpose: Rescue stent (RS) is an accepted rescue option after failed mechanical thrombectomy (MT) for acute ischemic stroke due to intracranial atherosclerotic stenosis (ICAS)-related large vessel occlusion (LVO). However, the long-term outcomes (≥ 12 months) of RS have not yet been elucidated.

Materials And Methods: We retrospectively analyzed the data of 154 patients with RS for ICAS-related LVO, which were identified from prospectively maintained multicenter database of RS after MT failure, to assess good outcome (mRS 0-2), mortality, stroke recurrence, symptomatic intracranial hemorrhage (SICH) and stent patency.

Integrated molecular and functional characterization of the intrinsic apoptotic machinery identifies therapeutic vulnerabilities in glioma.

Genomic profiling often fails to predict therapeutic outcomes in cancer. This failure is, in part, due to a myriad of genetic alterations and the plasticity of cancer signaling networks. Functional profiling, which ascertains signaling dynamics, is an alternative method to anticipate drug responses.

Possibilities and Limitations in Substituting anti-Drug Antibody Titers with Signal-to-Noise Ratios: A Comprehensive Comparison Using Two Clinical Trial Datasets of Adalimumab.

Immunogenicity assessment is vital in clinical trials and is measured through a multi-tiered approach (screening, confirmatory and titer assays). However, recent studies have suggested that titer results could be reported from ADA signal-to-noise ratios (S/N ratios=sample mean signal/negative control mean signal). More data analysis using two clinical trials of adalimumab: SB5-1003 (single-dose, healthy participants) and SB5-4001 (multiple-dose, interchangeability study, patients with plaque psoriasis), therefore, is indispensable whether substituting ADA S/N ratio as an alternative way of reporting titer results has no impact on interpretation on clinical outcome.

Small-molecule disruption of androgen receptor-dependent chromatin clusters.

Sustained androgen receptor (AR) signaling during relapse is a central driver of metastatic castration-resistant prostate cancer (mCRPC). Current AR antagonists, such as enzalutamide, fail to provide long-term benefit for the mCRPC patients who have dramatic increases in AR expression. Here, we report AR antagonists with efficacy in AR-overexpressing models.

Thinking politically about intersectoral action: Ideas, Interests and Institutions shaping political dimensions of governing during COVID-19.

Our paper examines the political considerations in the intersectoral action that was evident during the SAR-COV-2 virus (COVID-19) pandemic through case studies of political and institutional responses in 16 nations (Australia, Belgium, Brazil, Ethiopia, India, New Zealand, Nigeria, Peru, South Africa, South Korea, Spain, Taiwan, Thailand, Vietnam, UK, and USA). Our qualitative case study approach involved an iterative process of data gathering and interpretation through the three Is (institutions, ideas and interests) lens, which we used to shape our understanding of political and intersectoral factors affecting pandemic responses. The institutional factors examined were: national economic and political context; influence of the global economic order; structural inequities; and public health structures and legislation, including intersectoral action.

Protein Biomarkers of Adverse Clinical Features and Events in Sarcomeric Hypertrophic Cardiomyopathy.

Background: Hypertrophic cardiomyopathy (HCM) is a heterogeneous condition that can lead to atrial fibrillation, heart failure, and sudden cardiac death in many individuals but mild clinical impact in others. The mechanisms underlying this phenotypic heterogeneity are not well defined. The aim of this study was to use plasma proteomic profiling to help illuminate biomarkers that reflect or inform the heterogeneity observed in HCM.

Immunostimulatory Activity of a Mixture of , , , and in RAW264.7 Macrophages.

In this study, a mixture of , , , and (PPCRE) was investigated for their immuno-enhancing effects, as well as the molecular mechanism of PPCRE in RAW264.7 cells. PPCRE dramatically increased nitric oxide (NO) and prostaglandin E (PGE) generation depending on the concentration while exhibiting no cytotoxicity.

Visual and patient reported outcomes provided by a refractive multifocal intraocular lens based on continuous transitional focus.

Purpose: To analyze the quality of vision of patients implanted bilaterally with the multifocal Precizon Presbyopic intraocular lens (IOL), as well as to evaluate the visual performance provided by the lens.

Setting: Vissum Miranza Alicante.

Design: Prospective multicenter study.

Clinical validity and precision of deep learning-based cone-beam computed tomography automatic landmarking algorithm.

Purpose: This study was performed to assess the clinical validity and accuracy of a deep learning-based automatic landmarking algorithm for cone-beam computed tomography (CBCT). Three-dimensional (3D) CBCT head measurements obtained through manual and automatic landmarking were compared.

Materials And Methods: A total of 80 CBCT scans were divided into 3 groups: non-surgical (39 cases); surgical without hardware, namely surgical plates and mini-screws (9 cases); and surgical with hardware (32 cases).

Adjuvant Pembrolizumab versus Observation in Muscle-Invasive Urothelial Carcinoma.

Background: Muscle-invasive urothelial carcinoma is an aggressive disease with high rates of relapse. Whether pembrolizumab as adjuvant therapy would be effective in patients with high-risk muscle-invasive urothelial carcinoma after radical surgery is unknown.

Methods: In this phase 3 trial, we randomly assigned patients, in a 1:1 ratio, to receive pembrolizumab at a dose of 200 mg every 3 weeks for 1 year or to undergo observation.