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Lenvatinib plus Pembrolizumab or Everolimus for Advanced Renal Cell Carcinoma
Lenvatinib plus either pembrolizumab or everolimus was compared with sunitinib as first-line therapy for advanced renal cell cancer. Progression-free survival was significantly longer with lenvatinib plus pembrolizumab than with sunitinib. Lenvatinib plus everolimus was also more effective than sunitinib, but the difference was smaller.
Journal Article
NTP security : a quick-start guide
\"Learn the risks associated with Network Time Protocol (NTP) security and how to minimize those risks in daily deployment. Disruption of NTP services can interrupt communication between servers on the network and take an entire network offline. Beyond disrupting communication, flaws in the NTP daemon itself can make servers vulnerable to external attack--attacks that often go unnoticed. NTP is being used more frequently in Distributed Denial of Service (DDoS) attacks. It is a User Datagram Protocol (UDP) with encryption schemes that are not often used or are poorly implemented, making it susceptible to spoofing. Despite all of the security challenges, the fact is that NTP is critical to most modern networks. It is one of those \"set it and forget it\" protocols that network administrators and even security professionals don't understand in depth. However, an attacker who does understand the security flaws can wreak havoc on an insecure network. NTP Security: A Quick-Start Guide provides a deeper understanding of the protocol itself and how to deploy a strategy using the protocol throughout a network in a secure manner. Your security team will be able to provide better guidance to the system and network teams who will then be able to better manage the day-to-day implementation. This succinct resource offers practical guidance to an underserved topic (actually, not served at all). Coverage includes: an understanding of NTP and the importance of time synchronization in modern networks; issues in NTP security, including an analysis of NTP traffic; a review of the vulnerabilities and flaws in the protocol; practical solutions for securing NTP and building a robust infrastructure; effective alternatives to NTP\"--Back cover.
Book
I-TEP: A Simple and Affordable Method to Measure Permeability in Reconstructed Tissues Combined with DAMO–TSC-Based Urea Assay
Trans-epithelial permeability is a critical functional parameter for reconstructed tissues, particularly in genitourinary tissue engineering, where urine leakage must be avoided. Although Franz diffusion cells are considered the gold standard for permeability measurements, their cost and limited accessibility restrict their widespread use. In parallel, the reliable quantification of urea in culture media remains challenging due to protein interference and assay cost. The Inexpensive Trans-Epithelial Permeability (I-TEP) test is a simple and a low-cost Franz-like permeability system which can be combined with an optimized diacetyl monoxime–thiosemicarbazide (DAMO–TSC) colorimetric urea assay. I-TEP system relies on readily available laboratory components to create physically separate donor and receiver compartments, with the tissue acting as the sole diffusion interface. The DAMO–TSC assay was optimized through systematic evaluation of deproteinization, incubation time, storage conditions, and serum interference. The I-TEP test showed a strong correlation with conventional Franz diffusion cells when testing similar tissue samples. Deproteinization was identified as a mandatory step for accurate urea quantification in serum-containing media. The combined approach was successfully applied on engineered genitourinary tissues, demonstrating sensitivity to tissue maturation and cellular composition. This protocol provides a proof of concept for an affordable, robust, and autonomous method for routine permeability assessment, bridging the gap between costly commercial systems and high-throughput experimental needs.
Journal Article
Collection and Lipidomic Analysis of Murine Knee Synovium and Infrapatellar Fat Pad
Intra-articular soft connective tissues such as synovium and adipose tissue play a crucial role in governing joint homeostasis and disease progression in various forms of arthritis. In the knee, like many synovial joints, adipose tissue forms an integrated anatomic and functional unit with the joint-lining synovium, and the most prominent adipose depot is the infrapatellar fat pad (IFP). With growing evidence that lipid profiles in the synovium–IFP unit shift during progression of joint diseases like osteoarthritis (OA), there is strong impetus for consistent tissue collection approaches and reproducible subsequent lipid characterization. Here, we present a standardized dissection and low-input untargeted lipidomics workflow optimized for mouse knee synovium and IFP, to enable comprehensive lipid profiling. Synovium/IFP from multiple joints are pooled to increase input mass and guarantee robust lipid yield, followed by lipid extraction and high-resolution liquid chromatography-mass spectrometry (LC–MS) acquisition for global, untargeted lipidomic profiling. The analysis workflow encompasses robust feature detection, accurate lipid annotation, data transformation and normalization. These steps enhance comparability across samples, particularly those with low input amounts, while minimizing technical variance and batch effects. Using this approach, we detect a broad spectrum of lipid species spanning the major lipid categories. As expected for untargeted discovery, a subset of non-lipid species is also observed. This protocol provides a practical framework for robust, reproducible lipidomics in murine intra-articular soft tissues to support future disease-specific biomarker and drug target discovery in OA and other joint diseases.
Journal Article
A practical guide for characterization of novel CRISPR-Cas systems with Pro-CRISPR factors
The emergence of advanced genome editing technologies has revolutionized research in life sciences, offering an unprecedented way to uncover unknown biological functions and innovative therapeutic strategies. Among all genome editing tools, CRISPR-Cas-based technologies play a pivotal role in this revolution, particularly Class 2 effectors such as Cas9 and Cas12, owing to their high efficacy and ease of programmability. With the advancements in genome sequencing and metagenomics, an increasing number of novel CRISPR-Cas systems have been discovered, including those found in extreme environments and viruses. Furthermore, recent studies have revealed an unexpected role of non-Cas accessory genes, such as the Tn7-like transposon and Pro-CRISPR factors (Pcr), in conferring additional functionalities to the CRISPR system, providing new insights into the understanding of CRISPR-mediated bacterial immunity and advancing the development of genome editing technologies. Therefore, it is essential to develop comprehensive methods for characterizing the Cas proteins and Pro-CRISPR factors with a growing diversity. In this protocol, we provide a method encompassing protein purification, biochemical characterization, validation of protein-protein interactions, and preliminary
functional assays in bacteria for Cas nuclease and its associated Pro-CRISPR factor. We hope this protocol will not only assist in the characterization of the CRISPR-Cas system, but also provide valuable guidance for the characterization of other nucleases or nucleic acid modification systems.
Journal Article
The stack
\"An accessible, cutting-edge introduction to the \"stack,\" the foundation beneath everything that's Internet-connected\"-- Provided by publisher.
BOOK
Rapid identification of antigen-specific TCRs for cancer immunotherapy
T cell receptors (TCRs) can recognize peptides presented by major histocompatibility complex (MHC) molecules, referred to as HLA in humans, which enables the targeted eradication of tumor cells expressing specific antigens. In recent years, TCR-engineered T cell (TCR-T) cell therapy has demonstrated substantial advancements in clinical trials targeting solid tumors. Notably, in August 2024, the U.S. FDA approved the first TCR-T drug for the treatment of advanced synovial sarcoma, representing a pivotal milestone in the field. For the development of TCR-T therapy, identifying tumor-associated antigen epitopes and high-functional TCRs are critical. Here, we present a comprehensive protocol outlining the process of identification of immunogenic epitopes and the efficient screening of antigen-specific TCRs from HLA transgenic mice. Additionally, the protocol encompasses methodologies for TCR-T cell preparation and their functional evaluation
. These approaches provide a robust framework for advancing the development of tumor-specific TCRs and fostering the clinical translation of TCR-T therapies.
Journal Article