Animal Tissue Samples Research Articles

Direct and cost-effective method for histone isolation from cultured mammalian cells

Histones are an essential part of nucleosomes that regulate chromatin structure and function. Histone exchanges and modifications represent a scaffold for DNA transcription, repair, and replication. Studying histones and histone code is an important and fast-developing branch of epigenetic science. Here we propose a fast, efficient, and versatile assay for nucleosomal histone isolation from mammalian cells, without the use of acids or high salt solutions which are common for other histone isolation techniques. All components used in the protocol are common and inexpensive laboratory chemicals. The protocol has been evaluated on six commonly used cell lines and two animal tissue samples. The mild extraction conditions preserve delicate histone epigenetic changes, allowing its downstream analyses. We have demonstrated the assays’ successful application during changes in the transcriptional activity of histone genes, cell cycle transitions, and DNA-damaging conditions. Histone fractions, obtained by the protocol, can be used for further applications, such as electrophoresis, immunoblot, and mass spectrometry. Therefore, the new proposed nucleosomal histone isolation method is sensitive, specific, and suitable for downstream applications of various kinds.

Rapid quantification of 50 fatty acids in small amounts of biological samples for population molecular phenotyping.

Efficient quantification of fatty-acid (FA) composition (fatty-acidome) in biological samples is crucial for understanding physiology and pathophysiology in large population cohorts. Here, we report a rapid GC-FID/MS method for simultaneous quantification of all FAs in numerous biological matrices. Within eight minutes, this method enabled simultaneous quantification of 50 FAs as fatty-acid methyl esters (FAMEs) in femtomole levels following the efficient transformation of FAs in all lipids including FFAs, cholesterol-esters, glycerides, phospholipids and sphingolipids. The method showed satisfactory inter-day and intra-day precision, stability and linearity (R2 > 0.994) within a concentration range of 2-3 orders of magnitude. FAs were then quantified in typical multiple biological matrices including human biofluids (urine, plasma) and cells, animal intestinal content and tissue samples. We also established a quantitative structure-retention relationship (QSRR) for analytes to accurately predict their retention time and aid their reliable identification. We further developed a novel no-additive retention index (NARI) with endogenous FAMEs reducing inter-batch variations to 15 seconds; such NARI performed better than the alkanes-based classical RI, making meta-analysis possible for data obtained from different batches and platforms. Collectively, this provides an inexpensive high-throughput analytical system for quantitative phenotyping of all FAs in 8-minutes multiple biological matrices in large cohort studies of pathophysiological effects.

Comparative Study of Various Procedures for Extracting Doxorubicin from Animal Tissue Samples

This article presents a comparative study of selected deproteinization-, liquid–liquid-extraction- (LLE), and solid-phase-extraction (SPE)-based procedures for the isolation of doxorubicin (DOX) and daunorubicin (DAU) as an internal standard (IS) from rat tissue samples. During the experiments, all samples were analyzed via liquid chromatography coupled with fluorescence detection (LC-FL), with analytes being monitored at excitation and emission wavelengths of 487 and 555 nm, respectively. The absolute recoveries of the sample-preparation procedure were then calculated and compared, and the advantages and disadvantages of each approach were considered in depth. Ultimately, SPE with hydrophilic–lipophilic balanced (HLB) sorbents was selected as the most effective extraction procedure as it enabled the absolute recovery of DOX from tissue samples at a level of 91.6 ± 5.1%. Next, the selected HLB-SPE protocol was coupled with LC-FL separation and the resultant method was validated according to FDA and ICH requirements. The validation data confirmed that the developed procedure met all required criteria for bioanalytical methods, with a limit of detection (LOD) and limit of quantification (LOQ) of 0.005 µg/g and 0.01 µg/g, respectively. Finally, the developed protocol was successfully tested on various rat tissues enriched with DOX, confirming its potential as an interesting alternative to previously reported protocols for pharmacokinetic studies and clinical investigations aimed at analysis of the level and biodistribution of DOX in tissue samples after systemic administration of this drug.

The Effect of the Inclusion of Different Concentrates in Feed Rations on the Contents of Tocopherols, β-Carotene and Retinol in the Livers and Longissimus dorsi Muscles of Farm-Raised Fallow Deer (Dama dama L.).

This study analyzed the chemical composition and contents of β-carotene, tocopherols and retinol in the longissimus dorsi (LD) muscles and livers of fallow deer (Dama dama L.) fed diets based on hay containing different concentrates (hay and oats—HO; hay, oats and protein concentrate—HOP; hay and pellets—HP). The provitamin and lipophilic vitamin contents in the samples of feed and animal tissues were determined by high-performance liquid chromatography (HPLC). The contents of retinol were nearly two-fold higher (p < 0.002) in the livers and many-fold higher in the LD muscles of fallow deer fed the HOP and HP diets. β-carotene was not identified in the livers or LD muscles of fallow deer. The concentrations of α-tocopherol and total tocopherols were higher in the livers and LD muscles of the animals that received the HO and HP diets, but the noted differences were not significant. The inclusion of various concentrates in the feed rations had no influence on the proximate chemical compositions of the livers or LD muscles of fallow deer.

Effects of Molybdenosis on Antioxidant Capacity in Endangered Przewalski's Gazelles in the Qinghai Lake National Nature Reserve in the Northwestern China.

The purpose of this study is to investigate the effects of molybdenosis on antioxidant capacity in endangered Przewalski's gazelles (Procapra przewalskii) in the animal rescue center in the Qinghai Lake National Nature Reserve in Northwestern China. Ten P. przewalskii in molybdenosis were selected and treated orally with copper sulfate (CuSO4) at a dose of 20g/animal/5days for 20days. Ten healthy P. przewalskii were also selected and allocated to healthy pastures for 20days. Samples of soil, forage, and animal tissue were collected. The values of mineral and hematological parameters were measured. Results showed levels of molybdenum (Mo) in soil and forage were significantly higher than those in healthy ranches (P < 0.01). The Mo content of blood and hair in gazelles from the rescue center was also significantly higher than those in the healthy ranches animals (P < 0.01). The copper (Cu) contents in blood and hair from the rescue center were significantly lower than those from the healthy pasture (P < 0.01). Hemoglobin (Hb) and red cell volume (PCV) in the gazelles from the animal rescue center were significantly lower than those from animals in healthy ranches (P < 0.01). Aspartate aminotransferase (AST), lactate dehydrogenase (LDH), phosphocreatine kinase (CPK), and alkaline phosphatase (ALP) were significantly higher than those from gazelles in healthy ranches (P < 0.01). The antioxidant capacity in gazelles from the animal rescue center was significantly lower than that of gazelles in healthy ranches. Supplementing CuSO4 significantly decreased the Mo content of blood and cured molybdenosis in gazelles. In summary, the Mo content of soil and forage was very excessive for gazelles in the animal rescue center. The antioxidant capacity of P. przewalskii has been seriously affected by molybdenosis.

MiR-450b promotes cell migration and invasion by inhibiting SERPINB2 in oral squamous cell carcinoma.

microRNA-450b (miR-450b) plays an important role in cancer progression; however, its function in oral squamous cell carcinoma (OSCC) remains largely unknown. This study aimed to investigate the action mechanisms of miR-450b in OSCC. OSCC animal model was established via continuous induction with single-drug 7, 12-dimethylbenzo[a]anthracene (DMBA). Animal tissue samples were pathologically typed using haematoxylin-eosin (HE) staining. The Cancer Genome Atlas (TCGA) database was used to predict miR-450b and SERPINB2 expression in head and neck squamous cell carcinoma (HNSCC). qRT-PCR and Western blotting were used to detect gene and protein expression in OSCC tissue and cells, respectively. OSCC cell proliferation, growth, migration and invasion were detected using CCK-8, colony formation, transwell migration and matrigel invasion assays, respectively. Bioinformatic tools were used to predict miR-450b target genes. Dual-luciferase reporter assay was used to verify targeting between miR-450b and SERPINB2. Finally, small interfering RNA (siRNA) was used to reduce SERPINB2 expression to detect its effect on tumourigenesis. Four stages of OSCC carcinogenesis (normal oral epithelium, simple epithelial hyperplasia, dysplasia and OSCC) were identified. miR-450b was found to be overexpressed in OSCC animal samples, HNSCC samples and human OSCC cells. Upregulation of miR-450b significantly promoted OSCC cell proliferation, colony formation, migration and invasion, while its downregulation had the opposite effect. SERPINB2 was found to be a miR-450b target gene, and its expression was negatively correlated with miR-450b expression. Altering SERPINB2 expression effectively inhibited OSCC cell invasion, metastasis and epithelial-mesenchymal transition (EMT). miR-450b plays a key role in OSCC tumourigenesis by regulating OSCC cell migration, invasion and EMT via SERPINB2.

Development of a Highly Specific Fluoroimmunoassay for the Detection of Doxycycline Residues in Water Environmental and Animal Tissue Samples.

Doxycycline (DOX) and its metabolite residues in food and the environment pose a serious threat to human health and the ecological environment. In this work, a novel method, termed competitive fluoroimmunoassays (cFIA), based on monoclonal antibody (mAb) bio-conjugated CdSe/ZnS core-shell quantum dots (QDs), was developed for sensitive and rapid bioanalyses of DOX in natural water and commercial meats. After the optimization of the experimental conditions, 1 μg mL-1 of coating antigen and 0.5 μg mL-1 of QD-labeled mAb were used for the establishment of the cFIA. With this assay, the 50% inhibition concentration was found to be 0.35 ng mL-1 of DOX in phosphate-buffered saline samples, and the limit of detection was 0.039 ng mL-1 with minor cross-reactivity to other tetracycline members. The recoveries from natural water and commercial meats spiked with DOX concentrations of 10-600 ng mL-1 were 81.3-109.8%, and standard deviation were all below 12%. Levels measured with the QD-cFIA for thirty authentic samples were confirmed by high-performance liquid chromatography with good correlations. These results indicate that QD-cFIA is sultable for the rapid and quantitative detection of DOX residue in environmental and food samples.

Isotopic signature in isolated south-western populations of European brown bear (Ursus arctos)

Stable isotope analysis of animal tissue samples is increasingly used to study the trophic ecology of target species. The isotopic signatures respond to the type of diet, but also to the environmental conditions of their habitat. In the case of omnivorous, seasonal or opportunistic feeding species, the interpretation of isotopic values is more complex, as it is largely determined by food selection, either due to individual choice or because of availability. We analysed C and N isotopes in brown bear (Ursus arctos) hair from four isolated populations of south-western Europe (Cantabrian, Pyrenees, Central Apennines and Alpine) accounting for the geographical and climatic differences among the four areas. We found inter-population differences in isotopic signatures that cannot be attributed to climatic differences alone, indicating that at least some bears from relatively higher altitude populations experiencing higher precipitation (Pyrenees) show a greater consumption of animal foods than those from lower altitudes (Cantabrian and Apennines). The quantification of isotopic niche space using Layman’s metrics identified significant similarities between the Cantabrian and Central Apennine samples that markedly differ from the Pyrenean and Alpine. Our study provides a baseline to allow further comparisons in isotopic niche spaces in a broad ranged omnivorous mammal, whose European distribution requires further conservation attention especially for southern isolated populations.

Evaluation of a single dose of intra-testicular insulin treatment in heat-stressed mice model.

Insulin plays important role in testicular functions such as germ cell proliferation and steroidogenesis, despite its conventional role as a hypoglycaemic agent. It is also well known that testicular activity is severely get affected by heat stress and heat stress induces testicular pathogenesis. The effect of insulin on heat-induced testicular impairment has not been investigated. Thus, it is hypothesized that insulin might modulate testicular activity in a heat-stressed model. Experimental mice were separated into 4 groups; the first group was the normal control (CN), and the second group was subjected to heat stress (HS) by submerging the lower body part in a thermostatically controlled water bath maintained at 43°C for 15 min. The third and fourth groups were treated with a single dose of intra-testicular insulin (0.6IU/mice) before and after heat stress. Animal tissue samples were collected after 14 days of heat treatment. Insulin treatment did not improve the sperm parameters; however, both insulin pre and post-treatment improved the markers of spermatogenesis such as Johnsen score, germinal epithelium height and the number of stages VII/VIII. The histoarchitecture of testis also showed amelioration from heat-induced pathogenesis in the insulin-treated groups. Insulin treatment has also increased the proliferation of germ cells (increased PCNA and GCN), survival (Bcl2), and decreased apoptosis (active caspase-3). Furthermore, insulin treatment decreased MDA levels, without pronounced effects on the activities of antioxidant enzymes. Heat stress also decreased the circulating testosterone and oestrogen levels, and insulin treatment significantly increased oestrogen levels only. Although testosterone showed an increasing trend, it was insignificant. The expression of aromatase, AR, ER-α, and ER-β was down regulated by heat-stress and insulin treatment up regulated these markers. In conclusion, our results showed the amelioration of heat-induced testicular impairment by pre and post-intra-testicular insulin treatments. Insulin-associated improvements in the pre-and post-treatment groups suggested a preventive mechanism of insulin against heat stress in the testis.

P436: TARGETED DELIVERY OF T22-PE24-H6 TO CXCR4 POSITIVE CELLS FOR ACUTE MYELOID LEUKEMIA TREATMENT

Background: Acute myeloid leukemia (AML) is a malignant heterogeneous group of hematological diseases that originates from a hematopoietic progenitor with altered maturation capacity. Recently, considerable advances have been made in the development of targeted therapies for distinct molecularly defined subtypes. Although many AML patients initially respond to current treatment, the majority of them relapse. Therefore, new targeted therapies are needed to overcome drug resistance and improve treatment effectiveness. CXCR4 is a chemokine receptor that is expressed in over 20 cancer types, both hematologic cancers and solid tumors. CXCR4 receptor and its ligand CXCL12 are key mediators of the interactions between AML cells and the bone marrow (BM) microenvironment. Moreover, CXCR4 is highly expressed in around 50% of AML patients. Thus, we have generated a nanoparticle (T22-PE24-H6) that selectively delivers the exotoxin A from Pseudomonas aeruginosa to CXCR4+ leukemic cells. Aims: To assess the selectivity of T22-PE24-H6 cytotoxicity in CXCR4+ AML cell lines as well as in BM samples from AML patients. In addition, we evaluated the antineoplastic effect of the nanoparticle in a CXCR4+ AML disseminated mouse model. Methods: OCI-AML-3, MONO-MAC-6 and HEL cell lines were characterized for CXCR4 expression using flow cytometry and immunohistochemistry (IHC). The in vitro antineoplastic effect of T22-PE24-H6 was determined by cell viability assays. Competition assays with the CXCR4 antagonist AMD3100 were performed to demonstrate the CXCR4-dependent cytotoxicity of the nanoparticle. In vivo effect was assessed in NSG mice intravenously injected with 1x106 luminescent MONO-MAC-6 cells. Animals were treated with 5 μg T22-PE24-H6 or Buffer (166 mM NaCO3H pH 8) daily for 10 doses. Bioluminescence signal (BLI) was monitored to quantitatively measure and evaluate in vivo disease progression. Animal tissue samples were collected after mice euthanasia and used to analyze AML dissemination and toxicity. CXCR4 expression levels and T22-PE24-H6 effect were also determined in BM samples from 10 newly-diagnosed AML patients. Results: T22-PE24-H6 nanoparticle demonstrated a potent in vitro anticancer effect in MONO-MAC-6 cell line. The specific cytotoxic activity of the nanoparticle was CXCR4 dependent as demonstrated by performing competition assays with AMD3100. Furthermore, T22-PE24-H6 intravenous administrations showed significant BLI reduction in a CXCR4+ AML disseminated mouse model compared to buffer-treated mice. In addition, no differences between groups in mouse body weight, biochemical parameters or histopathological changes in normal tissues were detected. Finally, T22-PE24-H6 causes a significant cell viability reduction in AML patient samples with high CXCR4 expression. In contrast, the nanoparticle has no effect in AML patient samples with low expression of CXCR4. Summary/Conclusion: T22-PE24-H6 selectively kills AML cell lines and BM samples from AML patients with CXCR4 overexpression. The nanoparticle also induces a high antineoplastic effect in a CXCR4+ AML disseminated mouse model without detectable systemic toxicity. These data strongly support that T22-PE24-H6 may obtain therapeutic benefit for AML patients with high CXCR4 expression.

Measurement of Tetrahydrobiopterin in Animal Tissue Samples by HPLC with Electrochemical Detection-Protocol Optimization and Pitfalls.

Tetrahydrobiopterin (BH4) is an essential cofactor of all nitric oxide synthase isoforms, thus determination of BH4 levels can provide important mechanistic insight into diseases. We established a protocol for high-performance liquid chromatography/electrochemical detection (HPLC/ECD)-based determination of BH4 in tissue samples. We first determined the optimal storage and work-up conditions for authentic BH4 and its oxidation product dihydrobiopterin (BH2) under various conditions (pH, temperature, presence of antioxidants, metal chelators, and storage time). We then applied optimized protocols for detection of BH4 in tissues of septic (induced by lipopolysaccharide [LPS]) rats. BH4 standards in HCl are stabilized by addition of 1,4-dithioerythritol (DTE) and diethylenetriaminepentaacetic acid (DTPA), while HCl was sufficient for BH2 standard stabilization. Overnight storage of BH4 standard solutions at room temperature in HCl without antioxidants caused complete loss of BH4 and the formation of BH2. We further optimized the protocol to separate ascorbate and the BH4 tissue sample and found a significant increase in BH4 in the heart and kidney as well as higher BH4 levels by trend in the brain of septic rats compared to control rats. These findings correspond to reports on augmented nitric oxide and BH4 levels in both animals and patients with septic shock.

Dual-wavelength multimodal multiphoton microscope with SMA-based depth scanning.

We report on a multimodal multiphoton microscopy (MPM) system with depth scanning. The multimodal capability is realized by an Er-doped femtosecond fiber laser with dual output wavelengths of 1580 nm and 790 nm that are responsible for three-photon and two-photon excitation, respectively. A shape-memory-alloy (SMA) actuated miniaturized objective enables the depth scanning capability. Image stacks combined with two-photon excitation fluorescence (TPEF), second harmonic generation (SHG), and third harmonic generation (THG) signals have been acquired from animal, fungus, and plant tissue samples with a maximum depth range over 200 µm.

Perception of cultured meat among Sri Lankan community

Production of cultured meat involves synthesizing meat from animal tissue samples in a controlled environment using biotechnology. This study was directed to understand the perception of Sri Lankan consumer on cultured meat consumption. A structural survey was conducted in Sri Lanka using a probability sample of 14200 people from 11 districts and it was selected proportionately to the population in selected divisional secretariat divisions of these districts. The result of the survey revealed that 75% of the population was interested in accepting the cultured meat. The study showed that the meat consumers usually considered the health benefits mostly followed by nutritional properties, sensory properties especially the flavor and texture and meat price. Participants have understood the advantages of cultured meat in terms of animal welfare, environmental aspects, and food security of the country. Therefore, it can be concluded that there will be a substantial market for cultured meat, when it is introduced to the market. Educating people about intrinsic properties of cultured meat will also be important. In addition, the regression analysis showed that the acceptance level of cultured meat increased with the education level of consumers.

Protective Effects of Quercetin on Clothianidin-Induced Liver Damage in the Rat Model.

Clothianidin (CTD) is a member of the neonicotinoid group of insecticides. This study was performed to determine the effect of quercetin on clothianidin-induced liver injury (CTD) in rats. Rats were randomly assigned to a normal control (saline), a CTD control-treated group (20 mg/kg) every 3 days for 21 days, and CTD + quercetin-treated groups (2.5, 5, and 10 mg/kg) for 35 days intraperitoneally. Enzyme activity, including alanine aminotransferase (ALT) and aspartate aminotransferase (AST), was measured by spectrophotometry in serum samples by an automatic biochemical analyzer using commercial kits. Total antioxidant capacity (TAC), malondialdehyde (MDA), and nitrate-nitrite were measured in homogeneous liver tissue samples of animals. A significant increase in ALT and AST enzyme activity was observed in the CTD group in comparison with that of the control groups. In the clothianidin + quercetin (10 mg/kg) group, the ALT and AST enzyme levels decreased compared to the clothianidin control group significantly (P < 0.05). The MDA value of the liver increased in the clothianidin-treated group compared to that of the control groups (P < 0.05). Decreased tissue TAC level was observed in the CTD-treated group in comparison with that of the control groups (P < 0.05). The MDA level of the liver decreased in the clothianidin + quercetin (10 mg/kg) group compared to that of the CTD control group (P < 0.05). Quercetin significantly raised the level of TAC in the liver tissue of the clothianidin + quercetin (10 mg/kg) treated group compared to that of the clothianidin control group (P < 0.05). Liver nitrate-nitrite measurement showed a significant increase in the clothianidin group compared to that of the normal control group (P < 0.05). Nitrate-nitrite level in the liver was decreased in clothianidin + quercetin (10 mg/kg) compared to that of the clothianidin control group significantly (P < 0.05). Histopathological investigation revealed that contact to the CTD induced tissue disorganization and inflammatory cell infiltration, but minor histopathological alterations in the liver tissues of rats treated with CTD and quercetin (10 mg/kg) were detected.

ОПРЕДЕЛЕНИЕ ОСТАТОЧНЫХ КОЛИЧЕСТВ ХЛОРОФОСА ПОСЛЕ НАРУЖНОГО ПРИМЕНЕНИЯ ПРЕПАРАТА «АНТИОВОД»

The article represents data on adaptation and development of method for chlorophos analysis (active substance of agent «Antiovod») in organs and tissues of calves and cow milk by gas-liquid chromatography method. The method is based on extraction of chlorophos from samples of animal organs, tissues, organs and milk by acetone with addition of 1% acetic acid solution, evaporation of acetone-aqueous extract followed by reextraction into chloroform, chloroform evaporation, dissolution of residue in acetone and analysis of samples using gasliquid chromatography. The sensitivity of the method is 0.005 mg/kg (or mg/l). Residues of chlorophos in tissues and organs of calves were determined on days 5; 10; 15 and 21 post topical treatment with agent «Antiovod». The highest agent concentration was found in the first period of studies, i.e. at day 5. At day 21 post topical application of agent «Antiovod» chlorophos was not detected in any organ of calves. It was found that there was no release of chlorophos with milk in dairy cows treated with agent «Antiovod» at all tested time points, i.e. at days 1; 3; 5 and 10 post application. Based on the obtained data it is recommended to slaughter cattle for meat at day 21 post treatment with «Antiovod». It is recommended to use milk from cows treated with agent «Antiovod» without restrictions.

Impact of tick-borne Anaplasma phagocytophilum infections in calves of moose (Alces alces) in southern Norway.

The Gram-negative, obligate intracellular tick-transmitted pathogen Anaplasma phagocytophilum can cause acute febrile diseases in humans and domestic animals. The expansion of the tick Ixodes ricinus (Linnaeus, 1758) in northern Europe due to climate change is of serious concern for animal and human health. The aim of the present study was to investigate the impact of A. phagocytophilum infection in moose Alces alces (Linnaeus) calves by evaluating the carcass weights of infected and non-infected animals and examining animal tissues samples for co-infections with either species of Babesia Starcovici, 1893 or bacteria of the genus Bartonella. The carcasses of 68 free-ranging moose calves were weighed by hunters during the hunting seasons from 2014 to 2017 in two regions in southern Norway and spleen samples were collected. Anaplasma phagocytophilum was detected in moose sampled from locations infected with ticks with a prevalence of 82% (n = 46). The carcass weights of A. phagocytophilum-infected calves (n = 46) and non-infected (n = 22) calves were compared. Although the average weight of infected calves (45.6 kg) was lower than that of non-infected calves (46.5 kg), the difference was not statistically significant. Three different variants of the bacterium 16S rRNA gene were identified. The average weight of animals infected with variant I was 49.9 kg, whereas that of animals infected with variant III was 42.0 kg, but the difference was not statistically significant (p = 0.077). Co-infections of A. phagocytophilum with Bartonella spp. or with Babesia spp. were found in 20 and two calves, respectively. A triple infection was found in two calves. Sequence analysis of the 18S rRNA gene of Babesia-positive samples revealed the presence of Babesia cf. odocoilei (Emerson et Wright, 1970). Strains of Bartonella closely related to Bartonella bovis (Bermond, Boulouis, Heller, Laere, Monteil, Chomel, Sander, Dehio et Piemont, 2002) were identified based on phylogenetic analysis of the gltA and rpoB genes. The loss of body mass in moose calves in the tick-infected site was probably influenced by multiple factors.

Time-gated Raman spectroscopy for biomedical application under ambient or strong background light conditions

Many biomedical applications require measurements of Raman spectra of tissue under ambient lighting conditions. However, the background light often swamps the weaker Raman signal. The use of time-gated (TG) Raman spectroscopy based on a single photon avalanche diode (SPAD) operating in time-correlated single photon counting and near-infrared laser excitation was investigated for acquisition of Raman spectra and spectral images of biological tissue. The results obtained using animal tissue samples (adipose tissue and muscle) show that the time gating modality enables measurement of Raman spectra under background light conditions of similar quality as conventional continuous wave Raman spectroscopy in the absence of background light. Optimal suppression of the background light was observed for time gate widths of 300–1000 ps. The results also showed that TG Raman spectroscopy was able to detect subtle spectral differences required for medical diagnostics, such as differences in Raman spectra of cancer and normal tissue. While the current instrument required scanning of the grating in order to obtain full Raman spectra, leading to impractical times for multi-wavenumber Raman mapping, imaging time could be drastically reduced by spectral multiplexing (compressed detection) using digital micromirror devices or by using SPAD arrays.

Evaluation of lead (Pb) concentration in animal tissue, soil, and water samples from grazing pasture near mining industry in southern part of Mongolia

Lead (Pb) is a substance that can cause serious damage to the brain and kidneys, soften bones, adverse effect to the reproductive system, and can be fatal. Long-term exposure to the substance can damage not only children, but also adults' nervous systems. Lead does not belong into the category of toxic substances due to its effects on the human body. It is considered as a harmful substance. In this study, we analyzed lead concentration in liver and kidney of cattle, horse, sheep, and goat from the mining areas located in the Ulaanbadrakh, Zuunbayan, and Airag soums of Dornogovi province. As a result, the level of lead in the liver of sheep and goat Zuunbayan soum of Dornogovi province was slightly higher than in other soums. The content of heavy metals in water and soil samples did not exceed the international standard average.

The Novotroitsk tailing dump influence on the arsenic and heavy metals accumulation in living organisms

To determine the Novotroitsk gold-arsenic deposit dumps influence on the ecological situation, soil samples were taken from 4 wells drilled in the tailing dump territory. The Southern Urals species plant material typical Samples were collected within a 2 m radius from the wells. Wool, muscle tissue, liver, and kidney samples were taken from the European moles caught on the dump. The comparison objects were plants and animal tissues samples from background areas at a distance of > 15 km from the tailing dump. In the samples, the elements’ contents were determined, incl. arsenic and heavy metals by inductively coupled plasma mass spectrometry. Heavy metals increased content in the soil was found, and the average content of arsenic exceeded the maximum permissible concentration in soils by 1256 times. As and Hg large amounts were also found in plant samples. In the mole wool and tissues, the As accumulation was noted 181.3 - 273.9 times higher than the background values, Hg - 9.9 - 70.8 times and somewhat less pronounced - copper, lead and zinc. The research results are recommended to be taken into account when organizing reclamation and reclamation measures, as well as mining waste storage and disposal.

Effects of Se-Yeast on Immune and Antioxidant in the Se-Deprived Pishan Red Sheep.

The experiments were to study the effects of Se-yeast on immune and antioxidant in Selenium(Se)-deprived Pishan red sheep in Southern Xinjiang, China. The samples of soil, forage, and animal tissues were collected, and used for measuring mineral content, physiological parameter, and biochemical values. These findings showed that the Se contents in affected soil and forage were markedly lower than those from unaffected soil and forage (P < 0.01). Se in affected blood and wool were also extremely lower than those from healthy Pishan red sheep (P < 0.01). The hemoglobin, packed cell volume, platelet count, Glutathione peroxidase, and total antioxidant capacity in the affected Pishan red sheep were markedly lower than those from healthy ones too (P < 0.01). The levels of malondialdehyde, total nitric oxide synthase, and lipid peroxide in Pishan red sheep from affected pastures were extremely higher than those from healthy ones (P < 0.01). The levels of interleukin (IL)-1β, Interleukin-2, tumor necrosis factor-α and interleukin-6 from serum were markedly decreased in affected Pishan red sheep(P < 0.01). The Pishan red sheep in Se-deprived pasture were treated by orally with Se-yeast, the amount of Se in the blood markedly increased in treated animals. Meanwhile, the immune and antioxidant indicator was returned to the healthy values. Consequently, our findings were indicated that Se-deprived forage caused oxidative damage, and a serious threat to the immune function in animals. The Se-yeast is more effective in the Se-deficient Pishan red sheep including blood Se content, immune function and the antioxidant capacity.