scholarly journals Anesthesia of laboratory animals in manufacturing of diagnostic and preventive biomedicines

2020 ◽  
Vol 10 (1) ◽  
pp. 83-89
Author(s):  
A. P. Semakova ◽  
V. G. Germanchuk ◽  
V. V. Rogozhin ◽  
N. Y. Shavina ◽  
M. V. Ovchinnikova ◽  
...  
Keyword(s):  
Specific Activity ◽  
Heart Beat ◽  
Raw Material ◽  
Laboratory Animals ◽  
Additional Stress ◽  
Rabbit Serum ◽  
Hypotensive Activity ◽  
Depth Of Anesthesia ◽  
Inhalation Anesthesia ◽  
Immunoglobulin Preparations

Preparations such as XilaVet, Zoletil 100 as well as Aeranne (Isoflurane) are successfully applied for animal anesthesia in veterinary practice. We assessed a possibility of using parenteral narcosis with Zoletil 100 in combination with muscle relaxant Xila for producer-rabbits involved in manufacturing of natural rabbit serum subsequently deployed for production of diagnostic serum and immunoglobulin preparations. Administration of preparations into auricular vein is easy to do, while animals are sedated immediately allowing for safe fixation on restraining table and causing no additional stress for biomodels. This type of narcosis provides for expected depth of anesthesia and its maintenance until the end of blood-letting procedure. Parameters characterizing the state of cardiovascular system due to anesthetic products remained within the permitted limits. These preparations do not reduce heart beat rate allowing for collecting sufficient blood volumes. Application of inhalation anesthesia with Aeranne in laboratory animals provides for the specified depth of anesthesia and its maintenance until the end of the whole procedure. However, it requires specialized equipment and highly trained personnel with appropriate skills. Usage of Xila as a mono narcosis is not recommended as exhibits weak analgesic effects and strong hypotensive activity by decreasing quantity of collected blood volume. It was found that anesthetics such as Xila, Zoletil 100, Aeranne did not affect specific activity of immune sera in case of total dehematizing procedure. Moreover, antibody titers were not declined throughout entire observation (12 months) period and complied with the requirements of regulatory documentation. In addition, a feasibility of replacing old-fashioned anesthesia method with diethyl ether for a combination of safer contemporary preparations of Zoletil 100 and Xila was demonstrated while manufacturing tableted chemical cholera vaccine in experimental series with suckling rabbits used at diverse stages of raw material verification during surgical interventions. Xila, Zoletil 100, and Aeranne examined by us had no impact on the amount of blood obtained from donor-animals, immunological properties of the sera and ready-to-use diagnostic preparations. Such drugs were safe for all-age animals that comply with the requirements to anesthesia of animal biomodels and producer-animals in manufacturing of immunobiological preparations. Thus, our study allowed to conduct experiments with laboratory animals in a more humane manner.

scholarly journals Investigating Molecular Mechanisms of Immunotoxicity and the Utility of ToxCast for Immunotoxicity Screening of Chemicals Added to Food

2021 ◽  
Vol 18 (7) ◽  
pp. 3332
Author(s):  
Olga V. Naidenko ◽  
David Q. Andrews ◽  
Alexis M. Temkin ◽  
Tasha Stoiber ◽  
Uloma Igara Uche ◽  
...  
Keyword(s):  
Immune System ◽  
High Throughput ◽  
Environmental Protection Agency ◽  
High Throughput Screening ◽  
Molecular Mechanisms ◽  
Specific Activity ◽  
Laboratory Animals ◽  
In Vitro Assays ◽  
Toxicological Studies

The development of high-throughput screening methodologies may decrease the need for laboratory animals for toxicity testing. Here, we investigate the potential of assessing immunotoxicity with high-throughput screening data from the U.S. Environmental Protection Agency ToxCast program. As case studies, we analyzed the most common chemicals added to food as well as per- and polyfluoroalkyl substances (PFAS) shown to migrate to food from packaging materials or processing equipment. The antioxidant preservative tert-butylhydroquinone (TBHQ) showed activity both in ToxCast assays and in classical immunological assays, suggesting that it may affect the immune response in people. From the PFAS group, we identified eight substances that can migrate from food contact materials and have ToxCast data. In epidemiological and toxicological studies, PFAS suppress the immune system and decrease the response to vaccination. However, most PFAS show weak or no activity in immune-related ToxCast assays. This lack of concordance between toxicological and high-throughput data for common PFAS indicates the current limitations of in vitro screening for analyzing immunotoxicity. High-throughput in vitro assays show promise for providing mechanistic data relevant for immune risk assessment. In contrast, the lack of immune-specific activity in the existing high-throughput assays cannot validate the safety of a chemical for the immune system.

scholarly journals EXTRACTION AND PURIFICATION OF VEGETABLE PEROXIDASE: A REVIEW

2019 ◽  
Vol 16 (31) ◽  
pp. 692-703
Author(s):  
Aline HAAS ◽  
Cleiton VAZ ◽  
Aniela Pinto KEMPKA
Keyword(s):  
Enzymatic Activity ◽  
Specific Activity ◽  
Extraction Methods ◽  
Raw Material ◽  
Partial Purification ◽  
Buffer Solution ◽  
Degradation Of Dyes ◽  
Extraction And Purification ◽  
Wide Range ◽  
Purification Methods

Peroxidases are enzymes that catalyze the oxidation of various substrates, maintaining their enzymatic activity in wide ranges of pH and temperatures. These enzymes are used in processes for the degradation of dyes and phenolic compounds. Peroxidases are present in the tissues of several plants, and the search for new sources of this enzyme is necessary. This literature review aims to compile information about the extraction and/or purification of peroxidases contained in different plant tissues, presenting extraction methods, purification processes, enzymatic activities and their increments, according to the chemical and physical processes applied. Several plant sources can be raw material to obtain these enzymes, through different forms of extraction, where the processes of comminution predominate in the presence of buffer solution. For partial purification, are used precipitation with solvents (acetone and ethanol) and salts (ammonium sulfate) and centrifugation. For purification, chromatographic processes are used, in which molecular exclusion and affinity chromatography are prominent. It is concluded that there is a wide range of possibilities for obtaining the enzyme peroxidase from plants, with variability in the enzymatic activity when different extraction methods are applied. The purification methods used provide increases in the specific activity of the peroxidases.

scholarly journals Direct Absorption Method and Liquid Scintillation Counting for Radiocarbon Measurements in Organic Carbon from Sediments

Radiocarbon ◽  
2010 ◽  
Vol 52 (2) ◽  
pp. 794-799 ◽  
Author(s):  
Ionut Faurescu ◽  
Carmen Varlam ◽  
Ioan Stefanescu ◽  
Stela Cuna ◽  
Irina Vagner ◽  
...  
Keyword(s):  
Organic Carbon ◽  
Liquid Scintillation ◽  
Specific Activity ◽  
Potassium Dichromate ◽  
Isotopic Fractionation ◽  
Raw Material ◽  
Total Carbon ◽  
Wet Oxidation ◽  
Direct Absorption ◽  
Absorption Method

In this paper, we investigate a procedure for radiocarbon determination in forest soil and slurry from lake sediments. The total carbon in these samples can be both inorganic and organic. Inorganic carbon can be analyzed in a straightforward manner using the direct absorption method by sample acidification and CO2 capture. For organic carbon, we investigate a hybrid method using the wet-oxidation of organic carbon followed by direct absorption. To evaluate the wet-oxidation processes with potassium dichromate (K2Cr2O7) and potassium permanganate (KMnO4), we performed several experiments using different quantities of soil and sediments in order to establish the quantity of CO2 for each type of sample. The 2 methods offer comparable results for 14C-specific activity (about 0.234 ± 0.024 Bq/g C), values that are expected for these kinds of samples. We also investigated the possibility of isotopic fractionation occurring during CO2 production from raw material by measuring δ13C levels from samples and obtained CO2.

scholarly journals CROSS-PROTECTIVE PROPERTIES OF AN INFLUENZA VACCINE BASED ON HBC4M2E RECOMBINANT PROTEIN

2018 ◽  
Vol 63 (2) ◽  
pp. 68-76
Author(s):  
L. M. Tsybalova ◽  
L. A. Stepanova ◽  
M. A. Shuklina ◽  
S. V. Petrov ◽  
A. A. Kovaleva ◽  
...  
Keyword(s):  
Clinical Trials ◽  
Influenza A ◽  
Clinical Symptoms ◽  
Specific Activity ◽  
High Specific Activity ◽  
Influenza Viruses ◽  
Candidate Vaccine ◽  
Laboratory Animals ◽  
Core Antigen ◽  
Influenza A Viruses

One of the main problems in the area of influenza prophylaxis and pandemic prevention is the development of cross-reactive vaccines, i.e. vaccines directed against all subtypes of human influenza viruses. Such vaccines are being developed in many countries for more than 10 years. A number of vaccines are presently undergoing clinical trials. We created Uniflu candidate vaccine based on recombinant HBc4M2e protein consisting of 4 tandem-connected copies of the highly conserved ectodomain of M2 protein of the influenza A virus. These 4 copies were genetically fused to the carrier protein, namely hepatitis B core antigen. Commercially available Derinat was used as adjuvant in the candidate vaccine. Preclinical studies on laboratory animals (mice, ferrets) demonstrated that immunization with Uniflu leads to significantly higher level of specific immunoglobulins in the blood and bronchoalveolar lavages. Moreover, it produces immunoglobulins belonging to subtype IgG2a that is the most important mediator of antibody-dependent cytotoxicity. The vaccine under review stimulates the proliferation of T-lymphocytes, as well as the formation of CD4+ and CD8+ T-cells synthesizing ɣ-IFN. When infected with the lethal doses (5 LD50) of influenza A viruses of the subtypes H1N1, H2N2, H3N2, and H1N1pdm09, immunized animals typically developed mild form of illness. This kept them alive in 90-100% of cases, which demonstrated almost complete protection from death. Replication of the virus in the lungs of immunized mice was reduced by 1.8-4.8 log10. High immunogenicity of the vaccine, and reduced clinical symptoms following experimental infection, were demonstrated in ferrets as well. The developed recombinant vaccine Uniflu has high specific activity and cross-protection. Uniflu can be proposed as pre-pandemic vaccine, provided that it passes clinical trials.

scholarly journals Electrocorticography based monitoring of anaesthetic depth in mice

2021 ◽  
Author(s):  
Dominik Schmidt ◽  
Gwendolyn English ◽  
Thomas Gent ◽  
Mehmet Fatih Yanik ◽  
Wolfger von der Behrens
Keyword(s):  
Feature Extraction ◽  
Data Quality ◽  
Animal Welfare ◽  
Laboratory Animals ◽  
Gradient Boosting ◽  
Depth Of Anesthesia ◽  
Depth Of Anaesthesia ◽  
Isoflurane Anaesthesia ◽  
Anaesthetic Depth ◽  
Real Time Tracking

To improve animal welfare and data quality and reproducibility during research conducted under anaesthesia, anaesthetic depth in laboratory animals must be precisely monitored and controlled. While a variety of methods have been developed to estimate the depth of anaesthesia in humans, such tools for monitoring anaesthetic depth in laboratory animals remain limited. Here we propose an epidural electrocorticogram-based monitoring system that accurately tracks the depth of anesthesia in mice receiving inhalable isoflurane anaesthesia. Several features of the electrocorticogram signals exhibit robust modulation by the concentration of the administered anesthetic, notably, corticocortical coherence serves as an excellent indicator of anaesthetic depth. We developed a gradient boosting regressor framework that utilizes the extracted features to accurately estimate the depth of anaesthesia. Our method for feature extraction and estimation is conducted with a latency of only ten seconds, establishing a system for the real-time tracking of anaesthetic depth in mice.

scholarly journals Study of toxicity of arsenic-containing compounds isolated from brown algae Saccharina japonica in laboratory animals

Trudy VNIRO ◽  
2020 ◽  
Vol 181 ◽  
pp. 223-234
Author(s):  
L.S. Abramova ◽  
◽  
V.V. Gershunskaya ◽  
A.V. Kozin ◽  
D.A. Bondarenko ◽  
...  
Keyword(s):  
Brown Algae ◽  
Inorganic Arsenic ◽  
Laminaria Japonica ◽  
Saccharina Japonica ◽  
Raw Material ◽  
Toxic Effects ◽  
Laboratory Animals ◽  
Arsenic Content ◽  
Total Arsenic ◽  
Arsenic Compounds

The ability of various marine organisms, especially algae and invertebrates, to accumulate arsenic in high concentrations can pose a threat to public health when consumed. It is known from the literature that inorganic arsenic compounds (arsenites and arsenates) are the most toxic, in comparison with methylated forms of the element, and especially with complex organic compounds (arsenobetain, arsenocholine, tetramethylarsonium, arsenoriboses), which are considered non-toxic for live organisms. Monitoring of safety indicators of aquatic biological resources in the main commercial basins of the Russian Federation has shown that the most common excess of total arsenic content is characteristic for algae. According to TR CU 021/2011, the total arsenic content in algae should be 5 mg / kg and the established norm without separation of organic and inorganic arsenic compounds creates a barrier to the rational use of seafood. In this regard, the justification of the norms for the content of inorganic arsenic in algae and the assessment of their toxicity is a very urgent problem. Study of the samples of commercial brown algae Saccharina (=Laminaria) japonica and its derivates with ICP-MS, HPLC–MS-ISP methods, the maximum permissible level of arsenic was found to be exceeded, but the most toxic inorganic forms made up from 6 to14 % of the total amount of arsenic in the raw material. Acute toxicity on laboratory animals (rats) was studied and the absence of toxic effects was shown when an oral suspension containing high doses of arsenic was administered. Repeated administration of the same substances to laboratory mice of the CD 1 line has shown no toxic effects even after multiple doses of arsenic isolated from algae.

Purification and Partial Characterization of Glyceraldehyde-Phosphate Dehydrogenase from Electric Organ of Electrophorus electricus (L.)

1998 ◽  
Vol 53 (5-6) ◽  
pp. 416-420 ◽  
Author(s):  
S. Giovanni-De-Simone ◽  
A. Hassón-Voloch ◽  
C. Batista-e-Silva ◽  
A. Nery-da-Matta
Keyword(s):  
Amino Acids ◽  
Specific Activity ◽  
Electric Organ ◽  
Fold Increase ◽  
Rabbit Serum ◽  
Rabbit Muscle ◽  
Electrophorus Electricus ◽  
Glyceraldehyde Phosphate Dehydrogenase ◽  
Sds Page ◽  
Final Yield

Abstract The glyceraldehyde-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) was purified to homogeneity from electric organ of Electrophorus electricus (L.) by a hydrophobic chroma­ tography method on deacetylcolchicine-Sepharose. The purification resulted in a 162 fold increase in specific activity of the GAPDH and final yield was approximately 37%. The purified enzyme showed a single band in SDS-PAGE, with an apparent molecular mass of 36 kDa. The purity of the colchicine-Sepharose isolated material was analysed by isoelectro-phocusing and immunoblotting using a heterologous rabbit serum anti-GAPDH. Sequence analysis of the 40-N-terminal amino acids, determined by Edman degradation, revealed its identity to other GAPDHs proteins being the largest number of identical amino acids to lobster (92.5%), rabbit muscle (85%) and human liver (80%) GAPDH.

scholarly journals Inhalation anesthesia equipment for rats with provision of simultaneous anesthetic and oxygen

2011 ◽  
Vol 26 (2) ◽  
pp. 140-143 ◽  
Author(s):  
Edinaldo Gonçalves de Miranda ◽  
Vinícius Pontes do Nascimento ◽  
Daniel Reis Waisberg ◽  
Manoel Wilkley Gomes de Sousa ◽  
Marcel Fernando Miranda Batista Lima ◽  
...  
Keyword(s):  
Rattus Norvegicus ◽  
Wistar Rats ◽  
Depth Of Anesthesia ◽  
Average Volume ◽  
Inhalation Anesthesia ◽  
Equipment Operation ◽  
Anesthesia Equipment ◽  
Corneal Reflex ◽  
Operation Duration ◽  
Entire Procedure

PURPOSE: To introduce a model of equipment for inhalation anesthesia in rats that offers better control of both flow and losses of ether during induction, maintenance, and recuperation. METHODS: The equipment consists of an air compressor with two outlets, a closed glass induction chamber, a glass reservoir for the anesthetic agent, a pediatric inhalation mask, a three-way stopcock, a Y-connector, and urinary catheters. Three hundred Wistar rats (Rattus norvegicus albinus) were given inhalation anesthesia. The evaluated parameters were equipment operation, duration of each phase of anesthesia, corneal reflex, muscular tonus, respiration during induction and maintenance, and volume of anesthesia. RESULTS: The average time taken for induction was 7.3 minutes; the average anesthetic recuperation time was 6.4 minutes. The amount of anesthetic used varied according to the weight of the animal, with the average volume of ether used being 6.5ml/hour. The availability of oxygen (room air) decreased the recuperation time and averted both respiratory depression and insufficient depth of anesthesia. CONCLUSION: The proposed equipment is practical, inexpensive, and allows for satisfactory control of anesthetic parameters during the entire procedure, making inhalation anesthesia in rats safe and essentially complication free.

scholarly journals Interleukin-6 mRNA and protein increase in vivo following induction of acute thrombocytopenia in mice

Blood ◽  
1991 ◽  
Vol 77 (2) ◽  
pp. 286-293 ◽  
Author(s):  
LH Cox ◽  
T Downs ◽  
K Dagg ◽  
J Henthorn ◽  
SA Burstein
Keyword(s):  
Specific Activity ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Experimental Animals ◽  
Steady State Levels ◽  
Phosphate Buffered Saline ◽  
Protein Serum ◽  
Significant Increment

Abstract Induction of experimental thrombocytopenia in rodents results in the enhancement of megakaryocytic growth and differentiation. Interleukin-3 (IL-3) and IL-6, cytokines with a broad spectrum of biologic activities, stimulate megakaryocytopoiesis in vitro. To determine if expression of these factors might increase in response to experimental thrombocytopenia, we measured steady-state levels of IL-3 and IL-6 mRNA following rabbit antiplatelet serum (APS) injection. Groups of mice were injected intravenously with 0.2 mL APS while control animals received rabbit antilymphocyte serum (ALS), normal rabbit serum (NRS), or phosphate-buffered saline (PBS). At various times up to 72 hours after injection mice were exsanguinated and splenectomized. Platelet counts in the experimental animals were less than 12% of controls. Splenic RNA was hybridized in solution to 32P-UTP-labeled cRNA probes for IL-3 and IL-6. RNase-resistant hybrids were resolved on denaturing gels and visualized autoradiographically. IL-3 hybrids were undetectable at all time points tested, irrespective of the film exposure time or specific activity of the probe. Conversely, IL-6 hybrids were easily visualized and showed peak expression at 1.5 to 2.0 hours. By 3 hours, IL-6 mRNA had returned almost to the level of the controls. Similar results were observed in the bone marrow, although maximal IL-6 mRNA in that tissue was observed 4 hours following APS administration. To determine if this mRNA increment was associated with a concomitant increase in bioactive protein, serum was tested for its ability to stimulate IL-6-dependent B9 cells. At 1.75 hours following injection, experimental animals showed a small but significant increment in IL-6 activity compared with controls (200 +/- 30 U/mL IL-6 compared with 129 +/- 17 U/mL in ALS-injected controls, 106 +/- 17 U/mL in NRS-injected controls and 84 +/- 17 U/mL in PBS-injected controls). The data show that IL-6 mRNA and bioactive protein increase in response to acute immunothrombocytopenia, while no increment in IL-3 is detectable. These results suggest that IL-6 may play a role in the physiologic response to acute immunothrombocytopenia.

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