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Results for “RNA quantity”

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At least 19 recordsLinked to original sources
PubMed1977

[Trace changes in the quantity of RNA in neurons and neuroglia of the hypothalamic nuclei of the brain following short-term intermittent exposure to cold].

Adult albino rats were kept for 2 minutes at --20 degrees C, then for 5 minutes at 25 degrees C, this intermittent cooling being repeated 15 times. One hour after the end of the cooling, the RNA quantity per cell was determined by means of visible cytospectrophotometry to find the increase in the cytoplasm of medial preoptic area neurons and in the nuclei of their glial satellite cells as well as in the nuclei of the perineuronal glial cells of mammillary bodies. Two days after the rats had stayed at the room temperature, these RNA changey bodies the RNA content decreased. 3 days later, the RNA content in the whole neuron--neuroglia unit of the preoptic area returned to normal, while in the neurons and neuroglia of mammillary bodies augmented markedly: this augmentation was found as late as 15 days after the cessation of the cooling. 30 days after the end of the cooling, the RNA quantity in the mammillary body neurons returned to normal, whiel in the neuroglia of this hypothalamic nucleus decreased even lower than the normal level. The authors discuss the problem of a so-called "vegetative memory" and of the involvement of hypothalamic neuron--neuroglia units in the metabolism tranformation dealing with a consolidation of this memory.

Adaptation, Physiological↗
PubMed2026

Optimized Hot Phenol-Based RNA Extraction from Mycobacteria: A Robust Approach for Reliable Gene Expression Analysis.

Mycobacterium tuberculosis (Mtb) remains a major global health threat, underscoring the need for reliable transcriptomic studies to understand its biology and drug resistance mechanisms. Such analyses depend on obtaining high-quality, high-yield RNA. Although several RNA extraction methods are available, many require expensive reagents, large culture volumes, or specialized equipment, limiting their suitability for large-scale studies, particularly in resource-constrained settings. Here, an optimized Hot Phenol based RNA extraction method specifically tailored for mycobacteria is presented. The method uses minimal culture volume and commonly available reagents to consistently yield high-quality RNA suitable for high-throughput transcriptomic applications. RNA quantity and integrity were assessed by gel electrophoresis and RNA integrity analysis (RIN), and its suitability for downstream applications was confirmed by qPCR and Qubit 4. To benchmark the performance of the optimized method, a parallel RNA extraction using TRIzol and RNeasy under identical experimental conditions was carried out, including the same Mycobacterium species, culture volume, growth phase (logarithmic and stationary), and lysis conditions. This allowed a direct comparison of yield, quality, feasibility, and cost. The optimized Hot Phenol method demonstrated comparable or improved RNA yield and quality while significantly reducing reagent cost and dependence on specialized equipment. Owing to its efficiency, reproducibility, and affordability, this protocol provides a practical alternative for large-scale gene expression and transcriptomic studies in Mtb and other mycobacterial species.

RNA, Bacterial↗
PubMed1976

[Method of localization and evaluation of DNA quantities in bull spermatozoa].

The author attempted to evaluate the DNA quantities by means of a spectrophotometric method of UV light absorption. In a first publication data have been reported on the RNA quantities obtained by this method. This one concerns the data obtained on DNA. By this method, the manipulations on the spermatozoon are more dimple and sources of error are so discarded. It was so possible to get individual data on each spermatozoon, what is possible with chimical methods which give global quantities for a whole sample of semen. It is moreover possible to delimite the localization of DNA and to measure the amounts in each localization. The scale of the obtained data is large enough for information to several workers. The founded quantities and variations in data will be discussed in a following paper.

Animals↗
PubMedDate not supplied

[Participation of the genetic apparatus of the hippocampal and neocortical cells in the mechanisms of action of desglycine-arginine vasopressin on learning and memory processes in rats].

The paper deals with the influence of synthetic vasopressin analogue--desglycine-arginine vasopressin (DG-AVP) on the content of RNA and fractional composition of chromatin proteins in the tissue of neocortex and hippocampus of intact white rats and after establishing of two-way avoidance reflex. Administration of the peptide alone significantly increased RNA content in hippocampal tissue, injection of the peptide 10 min before conditioning did not lead to significant changes in RNA quantity as compared to that in animals in which the conditioned reflex was established against the background of saline administration. In neocortical tissue neither learning itself nor administration of DG-AVP alone was accompanied by significant changes in RNA content, while learning against the background of peptide injection significantly increased RNA in that structure. In hippocampal and neocortical tissues quantitative changes were observed in certain fractions of chromatin proteins in all animal groups studied.

Animals↗
PubMed1978

[Changes in the RNA concentration in neurons of the superior olivary complex in cats following monaural and binaural stimulation].

The nature of binaurally converging connections on the level of the superior olive nuclei was studied in cats anesthetized by the method of quantitative cytospectrophotometry. Significant increase in cytoplasmic RNA content in neurons of ipsi- and contralateral medial and lateral nuclei was observed after 2 hrs monaural stimulation nad 1.5 hrs binaural stimulation by rhythmical noise signals. The volume of functioning neurons of the nuclei studied was observed either to increase or not to differ from the control. The increase in RNA quantity both in ipsi-, and contralateral medial and lateral nuclei permits making the conclusion on the uniform distribution of binaurally converging connections on the neurons of these nuclei. Moreover, the data obtained enable us to assume that the accumulation of cytoplasmatic RNA occurs not only during excitation but also during inhibition.

Animals↗
PubMed1978

Chemical and conformational changes in chromosome regions being actively transcribed.

U.V. microspectrophotometry has been used to calculate quantities of nucleic acids and proteins of complete polytene chromosomal sets and specific regions of these chromosomes. It has been found that in chromosomes the ratio of DNA to proteins is approximately 1:4. This ratio however changes when specific regions are compared. The average ratio of DNA to proteins in a puffed region (2-48B4C5) increases to 1:16 in contrast to 1:6 from the same region but in non puffed state. At the same time the RNA quantity increases by a factor of 2. thermal denaturation profiles of formaldehyde fixed chromosomes show that the Tm of this region in puffed and non puffed state differ by 10 degrees C. Moreover these profiles suggest that a large fraction of histone-bound DNA is destabilized during puffing.

Animals↗
PubMed1991

Ponderable indices in the offspring and nucleic acids quantity in endocrine glands of rats exposed to real and sham VDU screens.

Adult female and male Imp: DAK rats were exposed beneath a real VDU screen (Erl groups) loaded with light-green characteristics due to computation of trigonometric functions in a loop program from a microcomputer. The control animals were exposed beneath a sham screen (Esh groups), being the front glass of vacuum devoid cathode ray tube covered with a transparent celluloid sheet printed with light-green characteristics. The difference in body weight of foetuses from autopsied Erl and Esh females was due mainly to a one-day delay in the fecundation of Esh females. In other groups of Erl females, a prolongation of the oestrous cycle was found in comparison to that of the Esh females. In both Erl females and Erl males, a nonsignificant decrease of weight of anterior pituitary and adrenals was found. RNA quantity was augmented in the anterior pituitary of Erl females and DNA quantity was diminished in the hypothalamus of Erl males. The obtained results suggest the suppression of hypothalamo-hypophyseal function in rats exposed to a real VDU screen.

Animals↗
PubMed2025

Systematic comparison of quantity and quality of RNA recovered with commercial FFPE tissue extraction kits.

BACKGROUND: FFPE tissue samples are commonly used in biomedical research and are a valuable source for next-generation sequencing in oncology, however, extracting RNA from these samples can be difficult the quantity and quality achieved can impact the downstream analysis. This study compared the effectiveness of seven different commercially available RNA extraction kits specifically designed for use with FFPE samples in terms of the quantity and quality of RNA recovered. METHODS: This study used 9 samples of FFPE tissue from three different types of tissue (Tonsil, Appendix and lymph node of B-cell lymphoma) to evaluate RNA extraction methods. Three sections of 20 µm of each sample were combined per sample. The slices were distributed in a systematic manner to prevent any biases. Each of the 7 commercially available RNA extraction kits were used according to manufacturer's instructions, with each sample being tested in triplicate resulting in a total of 189 extractions. The concentration, RNA quality score (RQS) and DV200 of each extraction was analysed using a nucleic acid analyser to determine the quantity and quality of the recovered RNA. RESULTS: This study found that despite processing the FFPE samples in the same standardized way, there were disparities in the quantity and quality of RNA recovered across the different tissue types. Additionally, the study found notable differences in the quantity of RNA recovered when using different extraction kits. In terms of quality, three of the kits performed better than the other four in terms of RQS and DV200 values. CONCLUSION: Though many laboratories have developed their own protocols for specific tissue types, using commercially available kits is still a popular option. Although these kits use similar processes and extraction procedures, the amount and quality of RNA obtained can vary greatly between kits. In this study, among the kits tested, while the Roche kit, provided a nearly systematic better-quality recovery than other kits, the ReliaPrep FFPE Total RNA miniprep from Promega yielded the best ratio of both quantity and quality on the tested tissue samples.

Paraffin Embedding↗
PubMed1990

Amplified RNA synthesized from limited quantities of heterogeneous cDNA.

The heterogeneity of neural gene expression and the spatially limited expression of many low-abundance messenger RNAs in the brain has made cloning and analysis of such messages difficult. To generate amounts of nucleic acids sufficient for use in standard cloning strategies, we have devised a method for producing amplified heterogeneous populations of RNA from limited quantities of cDNA. Whole cerebellar RNA was primed with a synthetic oligonucleotide containing the T7 RNA polymerase promoter sequence 5' to a polythymidylate region. After second-strand cDNA synthesis, T7 RNA polymerase was used to generate amplified antisense RNA (aRNA). Up to 80-fold molar amplification has been achieved from nanogram quantities of cDNA. The amplified material is similar in size distribution to the parent cDNA and shows sequence heterogeneity as assessed by Southern and Northern blot analysis. Specific messages for moderate-abundance mRNAs for actin and guanine nucleotide-binding protein (G-protein) alpha subunits have been detected in the amplified material. By using in situ transcription to generate cDNA, sequences for cyclophilin have been detected in aRNA derived from single cerebellar tissue sections. cDNA derived from a single cerebellar Purkinje cell also has been amplified and yields material that hybridizes to cognate whole RNA and mRNA but not to Escherichia coli RNA.

Actins↗
PubMed1979

Replication cycle of Newcastle disease virus in three host cells of different permissiveness.

Various degrees of permissiveness for NDV have been described in different cell lines. In the present work three systems were investigated: bovine kidney (MDBK) cells, chick embryo (CE) cells and mouse L cells producing 50 to 100 p.f.u., 2 to 10 p.f.u. and less than 0.1 p.f.u./cell, respectively. Analysis of the radioactive virus messenger RNAs (vmRNAs) accumulating in actinomycin D-treated cells throughout the infection cycle revealed that, except for the absence of one 18S vmRNA species in L cells, all vmRNA components were formed in the three cell types, although variations occurred in their total and relative amounts from one cell type to another. Kinetic studies of vmRNA synthesis confirmed the absence of one 18S vmRNA species in L cells and also showed that the labelling rate of this vmRNA component is higher in CE cells. Virus proteins synthesized in the infected cells were labelled with 14C-amino acids and analysed by polyacrylamide gel electrophoresis. All the major NDV polypeptides were formed as expected in both CE and MDBK cells but only traces were detected in L cells. In contrast in a cell-free translation system from wheat germ, all of the NDV major proteins were synthesized using RNA extracted from the three infected cell types. Moreover, the total radioactivity incorporated into NDV proteins was twice as great with CE cell RNA and five times as great with L cell RNA than with equivalent quantities of RNA from MDBK cells.

Animals↗