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Modified method for combined DNA and RNA isolation from peanut and other oil seeds.

Isolation of good quality RNA and DNA from seeds is difficult due to high levels of polysaccharides, polyphenols, and lipids that can degrade or co-precipitate with nucleic acids. Standard RNA extraction methods utilizing guanidinium-phenol-chloroform extraction has not shown to be successful. RNA isolation from plant seeds is a prerequisite for many seed specific gene expression studies and DNA is necessary in marker-assisted selection and other genetic studies. We describe a modified method to isolate both RNA and DNA from the same seed tissue and have been successful with several oil seeds …

A nucleotide-independent nitroxide probe reports on site-specific stereomeric environment in DNA.

In this report, stereospecific structural and dynamic features in DNA are studied using the site-directed spin labeling technique. A stable … ations in local DNA motions. This establishes R(p)- and S(p)-R5a as unique probes for investigating nucleic acids in a site- and stereospecific manner, which may aid studies of stereospecific DNA/protein interactions. In addition, weighted averages of individual R(p) and S(p) spectra match those of R5a attached to mixed diastereomers. This suggests that R5a lin …

Overexpression of transketolase-like gene 1 is associated with cell proliferation in uterine cervix cancer.

BACKGROUND: Tumor cells need large energy and nucleic acids to proliferate and grow. For most of their energy needs, cancer cells depend more on glycolysis. For most of their nucleic acids needs, cancer cells depend more on the nonoxidative pathway of the pentose phosphate pathway. Transketolase(TKT) is a crucial enzyme in the nonoxidative pathway of the PPP. METHODS: The real-time quantity PCR was used to determine the expression of transketolase gene family in uterine cervix cancer. Transketolase activity of cell was determined by using enzyme-linked method. Cell proliferation was detected …

Glomerular expression of toll-like receptor-9 in lupus nephritis but not in normal kidneys: implications for the amplification of the inflammatory response.

Toll-like receptors recognising self-derived nucleic acids may participate in the pathogenesis of autoimmune diseases. Following the description of an enhanced population of toll-like receptor-9 (TLR-9) expressing auto-antibody producing B lymphocytes in active lupus, we explored the expression of TLR-9 in the renal tissue of patients with lupus. TLR-9 expression was studied in the kidneys of 12 lupus and 10 control samples from macroscopically unaffected areas of patients with renal adenocarcinoma by immunohistochemistry. A semiquantitative score was assigned separately for tubular, interstit…

Mapping population immunity: How protease and antibody fragmentation shape wastewater findings.

Wastewater-based infection surveillance relies on nucleic acids detection, however incorporating antibody detection would allow deeper insights into population immunity. Antibodies serve as biomarkers of past infections and offer a longer timeline for surveillance compared to nucleic acids. However, the complex composition of wastewater presents challenges for the sensitivity of ELISA-based immunoassays. In our study, antibody profiles in stool and upstream wastewater samples were analyzed to evaluate the feasibility of detecting antigen-specific antibodies through immunoassays. In wastewater …

Accelerated CRISPR/Cas12a-Based Point-of-Care Diagnostics Through Critical Coupling Distance Control.

Prompt pathogen detection in resource-limited settings remains constrained by energy-intensive instrumentation and a shortage of trained personnel. The CRISPR/Cas12a-based diagnostic technology, despite its robustness as a promising tool, is constrained by suboptimal detection speed and sensitivity. Here we designed triblock DNA-mediated spherical nucleic acids (tSNA) that acts as a spatially confined reporter with critical coupling distances between substrates, enabling Cas12a protein to rapidly identify concentrated and stretched single-stranded substrates with size-matching intervals. Preci…

Cation Accumulation Drives the Preferential Partitioning of DNA in an Aqueous Two-Phase System.

Mixtures of polyethylene glycol (PEG) and dextran (Dex) represent a widely used class of aqueous two-phase systems (ATPS), with applications ranging from the purification of various biomolecules, such as nucleic acids, to the synthesis of protocells. A key feature underlying these applications is the selective accumulation of biomolecules within Dex-rich droplets in an aqueous PEG phase, but the physical origin of this partitioning remains unclear. Entropic interactions were long assumed to be the primary driving force; however, our systematic experiments using DNA of different lengths indicat…

Advances in nanomaterial-mediated CRISPR/Cas delivery: from lipid nanoparticles to vesicle-derived systems.

Gene and genome editing therapies are increasingly connected with nanomaterials, which protect and transport fragile nucleic acids and CRISPR/Cas systems through biological barriers safely and accurately. This review discusses how different nanocarriers, including lipid-based, polymeric, inorganic, and vesicle-derived systems, can improve delivery efficiency, cell targeting, endosomal escape, and intracellular movement for gene and genome editing. It summarizes findings from early clinical and preclinical studies, comparing several carrier types such as ionizable lipid nanoparticles, polymeric…

Reflections on the Principles of Fixation in Histochemistry and Cytochemistry: With a Special Focus on Immunohistochemistry and In Situ Hybridization.

In the field of histochemistry and cytochemistry (histocytochemistry), fixation is a critical process for preserving biological structures and enabling accurate analysis. Fixation methods, broadly categorized into precipitating and cross-linking techniques, stabilize biomolecules such as proteins, sugars (carbohydrates) and nucleic acids, although lipids often require specific handling due to the loss during a routine procedure. Traditional staining methods have evolved into advanced techniques like immunohistochemistry (IHC) and in situ hybridization (ISH), which allow for precise analysis of…

Development and characterization of an efficient, organic solvent-free capsular polysaccharide purification process from highly invasive Streptococcus pneumoniae serotype 22F.

Streptococcus pneumoniae serotype 22F is a significant concern in causing pneumococcal disease, and it is associated with increased rates of … -interaction chromatography (HIC) using Octyl Sepharose to effectively remove major impurities viz. nucleic acids, proteins, and cell wall common polysaccharides (C-Ps). This optimized process yielded high capsular polysaccharide recovery of approximately 65 % with molecular weight 840 kDa. The final CPS structural identity was confirmed by 1H NMR and it meets …

Evaluation of a Commercial Multiplex Real-Time PCR with Melting Curve Analysis for the Detection of Mycobacterium tuberculosis Complex and Five Nontuberculous Mycobacterial Species.

BACKGROUND: Accurate and timely diagnosis of mycobacterial infections, including Mycobacterium tuberculosis complex (MTBC) and nontuberculou … ollected from the Korea University Anam Hospital. RESULTS: The NeoPlex assay successfully detected nucleic acids in 87 of the 91 isolates (95.6%). Notably, it identified additional mycobacterial nucleic acids not detected by the LPA in eight isolates. These findings were confirmed via DNA sequencing. The assay had 100% sensitivity and specificity for M. intrac …

Establishment of a Direct PCR Assay for Simultaneous Differential Diagnosis of African Swine Fever and Classical Swine Fever Using Crude Tissue Samples.

African swine fever (ASF) and classical swine fever (CSF) are contagious swine diseases that are clinically indistinguishable from each other; hence, reliable test methods for accurate diagnosis and differentiation are highly demanded. By employing a buffer system suitable for crude extraction of nucleic acids together with an impurity-tolerant enzyme, we established a multiplex assay of real-time reverse-transcription polymerase chain reaction (rRT-PCR) for simultaneous detection of ASF virus (ASFV), CSF virus (CSFV) and swine internal control derived genes in a sample without the need for pr…

Иммуногенность вирусов Варичелла-Зостер Гликопротеин E, сформулированных с липидными нанопицерами и нуклеиновыми иммуностимуляторами в мышах.

Теоретически, вакцина от герпеса-зостера ShingrixTM могла бы использоваться в качестве вакцины от варицеллы, которая избегала бы риска разработки шинглов от вакцинации, однако стратегии смешивания в постели и ограниченное предложение адъютантного компонента QS21 сделали ее применение экономически непрактичным. С использованием липидных наночастиц (ЛНП), которые были утверждены УЛХ в качестве векторов для вакцин от острого острого дыхательного синдрома коронавируса 2, мы разработали серию вакцин, эффективно окапсулированных вирусом варицелла-зостер, гликпротеином Е (VZV-gE) и нуклеиновыми кисло

Совпадение в аннотации оригинала
Theoretically, the subunit herpes zoster vaccine ShingrixTM could be used as a varicella vaccine that avoids the risk of developing shingles … series of vaccines efficiently encapsulated with varicella-zoster virus glycoprotein E (VZV-gE) and nucleic acids including polyinosinic-polycytidylic acid (Poly I:C) and the natural phosphodiester CpG oligodeoxynucleotide (CpG ODN), which was approved by the FDA as an immunostimulator in a hepatitis B vaccine. Preclinical trial in mice showed that these LNP va …

The insufficiency of circulating miRNA and DNA as diagnostic tools or as biomarkers of treatment efficacy for Onchocerca volvulus.

Skin snip evaluation for onchocerciasis has insufficient sensitivity when skin microfilarial (mf) densities are low, such as following ivermectin treatment. Mf density is suitable for assessing microfilaricidal efficacy but only serves as an indirect indicator of macrofilaricidal activity. We assessed circulating nucleic acids from Onchocerca volvulus as an alternative to skin snips. We screened a plasma sample set of infected individuals followed up at four, 12 and 21 months after microfilaricidal (ivermectin, n = four), macrofilaricidal (doxycycline, n = nine), or combination treatment (n = …

Endosomal Toll-like receptors in clinically overt and silent autoimmunity.

Toll-like receptors (TLRs), first identified as pattern recognition receptors, are now recognized to serve as a key interface between innate and adaptive immunity. Systemic lupus erythematosus (SLE) is characterized by both continuous and cyclic stimulation of the innate and adaptive immune system by endogenous nucleic acids released from apoptotic or necrotic cells. TLR7 and TLR9 function as innate sensors of viral infection as their ligands are ssRNA and dsDNA, respectively. Recognition of self nucleic acids by endosomal TLRs in B cells and pDCs is thought to be an important step in the path…

Use of ethidium monoazide and propidium monoazide to determine viral infectivity upon inactivation by heat, UV- exposure and chlorine.

Despite the great sensitivity of PCR in monitoring enteric viruses in an aquatic environment, PCR detects viral nucleic acids of both infectious and noninfectious viruses, limiting the conclusions regarding significance for public health. Ethidium monoazide (EMA) and propidium monoazide (PMA) are closely related membrane impermeant dyes that selectively penetrate cells with compromised membranes. Inside the cells, the dye can intercalate into nucleic acids and inhibit PCR amplification. To assess whether EMA and PMA pretreatment is a suitable approach to inhibit DNA amplification from noninfec…

Mitochondrial DNA from archived tissue samples kept in formalin for forensic odontology studies.

BACKGROUND: Samples used for DNA isolation to be used for forensic odontology studies are often limited. The possibility to use tissue samples stored in formalin for a prolonged period, which contains nucleic acids of questionable quality, opens exciting possibilities for genetic and molecular biology studies useful in speciality of forensic odontology. AIM: The present study defines substantial modification of existing protocols for total genomic isolation including mitochondrial DNA and proves the utility of such obtained mitochondrial DNA in microsatellite analyses. METHODS: 50 dental tis…

Translational research in pediatrics II: blood collection, processing, shipping, and storage.

Translational research often involves tissue sampling and analysis. Blood is by far the most common tissue collected. Due to the many difficulties encountered with blood procurement from children, it is imperative to maximize the quality and stability of the collected samples to optimize research results. Collected blood can remain whole or be fractionated into serum, plasma, or cell concentrates such as red blood cells, leukocytes, or platelets. Serum and plasma can be used for analyte studies, including proteins, lipids, and small molecules, and as a source of cell-free nucleic acids. Cell c…

Development of a magnetic electrochemical bar code array for point mutation detection in the H5N1 neuraminidase gene.

Since its first official detection in the Guangdong province of China in 1996, the highly pathogenic avian influenza virus of H5N1 subtype ( … ce. Suggested experimental procedure allows obtaining further information from the redox signals of nucleic acids. Moreover, the used biosensor exhibits sequence specificity and low limits of detection of subnanogram quantities of target nucleic acids.

Protein arginine methylation rewires molecular interactions.

Protein arginine methyltransferases (PRMT1-9) catalyze monomethylation and dimethylation of arginine residues in RNA- and DNA-associated proteins. Protein arginine methylation can weaken or strengthen nucleic acid binding, recruit or exclude protein partners, and promote or suppress higher-order assembly, making its molecular consequences highly context dependent. Methylation preserves the cationic character of the guanidinium group, while altering hydrogen-bond donor topology, steric accessibility, and interaction geometry. These effects are particularly consequential in intrinsically disorde