Role and mechanism of Glut-1 and H+/K+-ATPase expression in pepsin-induced development of vocal cord leukoplakia

Purpose We investigated the role of Glut-1 and H+/K+-ATPase expression in pepsin-induced development of human vocal cord leukoplakia cells (HVCLCs). Next, we analyzed the relationship between Glut-1 and H+/K+-ATPase expression with the clinicopathological features of laryngeal carcinoma. Methods Glut-1 and H+/K+-ATPase expression levels in HVCLCs were determined after treatment with artificial gastric juice containing pepsin and laryngeal carcinoma tissues. Results Exposure to pepsin-containing artificial gastric juice significantly enhanced the migration and proliferation of VSCLCs in a time-dependent manner. The apoptotic rate of VSCLCs decreased over time after exposure to pepsin and reached a nadir on day 7 (p < 0.01). With increasing duration of exposure to pepsin, the proportion of VSCLCs in G0/G1 phase decreased and the proportions in the S and G2/M phases significantly increased (p < 0.05). After treatment with pepsin-containing artificial gastric juice, RT-PCR and Western blotting showed that the expression of Glut-1 and H+/K+-ATPase α, β significantly increased in HVCLCs compared to in the absence of pepsin (p < 0.05). The expression of Glut-1 and H+/K+-ATPase α, β gradually increased from vocal cord leukoplakia (VLC) to laryngeal carcinoma (p < 0.05). Lentivirus-mediated inhibition of Glut-1 expression in VCL significantly inhibited the cells’ migration and proliferation (p < 0.05) but enhanced their apoptosis (p < 0.05). Also, inhibition of Glut-1 expression resulted in an increased proportion of cells in G0/G1 phase and a significantly decreased proportion in G2/M phase (p < 0.05). Conclusions Elevated Glut-1 expression may promote the development of VCL by upregulating laryngeal H+/K+-ATPase expression to reactivate absorbed pepsin, thus damaging the laryngeal mucosa.

Trichinelloscopy of domestic and wild animal carcasses

The purpose of the research is analyze the localization of Trichinella sp. in animals muscle and to evaluate the methods of intravital and post-mortem diagnosis of trichinellosis in domestic, wild and game animals.Materials and methods. In order to prevent trichinellosis in human population and animals, life-time and post-mortem diagnosis methods for trichinellosis are widely used. Life-time diagnosis of trichinellosis is based on detection of specific antibodies in blood serum of sick animals. Modern immunological assays allow detecting specific antibodies (immunoglobulins J and M) at 10–12 days after infection. Enzyme-linked immunosorbent assay (ELISA) with fractionated antigen has the greatest real possibility of application for individual and mass seroepizootic studies of pigs and horses; ELISA is a highly sensitive and specific test. Veterinary and sanitary examination is conducted by methods of compressor trichinelloscopy and peptolysis (muscle tissue digested in artificial gastric juice). For the compression research method, in particular for pig carcasses, 2 samples of the diaphragmatic peduncles of 60 g each are taken. It is also possible to study samples from masticatory muscles, tongue, intercostal space or esophagus. Twelve sections are made from each sample (24 in total). A more sensitive and productive method is the digestion of muscle tissue using a set of diagnostic devices and instruments such as AVT. The method is based on peptolysis of crushed muscle tissue in technological reactors. Diagnosis of trichinellosis using such devices makes it possible to automate and mechanize all processes associated with the isolation of Trichinella larvae. The main application areas of the devices are meat processing factories, fur farms or veterinary and sanitary examination laboratories in the markets.Results and discussion. We presented data on the role of veterinary and sanitary examination for trichinellosis in susceptible animals as the core measure in the system of measures to prevent this infection. We analyzed indicators of diagnostic efficiency, performance and usability of methods of compressor trichinelloscopy and digestion of muscle tissue in artificial gastric juice. Factors of diagnostic efficiency, performance and usability of methods of compressor trichinelloscopy and digestion of muscle tissue in artificial gastric juice were analyzed. Information was given on localization of Trichinella larvae in various species of domestic and wild animals, optimal sampling sites and volumes of muscle tissue samples with the existing methods of examination for trichinellosis. To study fresh carcasses for trichinellosis at meat processing factories, the peptolysis method is recommended.

TRICHINELLOSIS OF THE STONE MARTEN (MARTES FOINA) IN CENTRAL RUSSIA

Over the past 5 years, research has been carried out in natural biocenosis on the stone marten (Martes foina) infected with the pathogen of trichinellosis in Central Russia. Biological material for research was collected in hunting farms, reserves and "green zones" of the Central Region of Russia (the Vladimir, Moscow, Nizhny Novgorod, Tula, Ryazan, Oryol and Bryansk regions) during the opening of amateur sports hunting for fur animals from 2017 to 2021. Diagnostics and isolation of helminth larvae from animal muscle tissue samples were performed by the compressor trichinelloscopy and digestion in artificial gastric juice. The degree of host invasion with helminth and invasion intensity were determined. A total of 35 animals were studied. As a result of studies, the infection rate of helminthozoonosis in animals was from 8.3 to 40.0%. The intensity of invasion was recorded between 2 and 39 larvae in 1 g of diaphragm muscle tissue. Trichinella invasion among martens was observed equally in both males and females. The largest percentage of the invasive form of the helminthiasis pathogen occurred in animals older than 2 years. The adverse situation of helminthozoonosis among fur animals was noted in the Ryazan region.

STUDY OF TRICHINELLA INFECTION IN WILD MAMMALS IN THE KURSK REGION

In the territory of the Kursk region, according to official statistics, trichinosis in humans is extremely rare, mainly in the form of imported cases. However, sometimes cases of the disease are detected among the population, which are associated with trichinella infection when eating meat of wild mammals living in the natural biocenoses of the Kursk region. In this regard, the purpose of our study was to study the infection of wild animals with Trichinella in the Kursk region. Trichinella larvae were identified by the method of compressor trichinelloscopy and the method of digestion of muscle tissue in artificial gastric juice. Capsule-forming larvae of Trichinella (Trichinella spp.) were found in the muscles of animals. Trichinella was found in the stone marten (10,5%), polecat (6,7%), red fox (5,9%), raccoon dog (22,2%) and white-breasted hedgehog (5,3%). Earlier, we conducted studies that showed that in the Kursk region, Trichinella parasitize not only in carnivores and insectivores, but also among rodents and wild boars. High rates of infection in predators indicate a wide spread of Trichinella in natural biocenoses of the Kursk region. This creates an actual risk for people to be infected with Trichinella. This risk is especially significant for family members of hunters, their relatives and friends.

Lactobacillus plantarum PMO 08 as a Probiotic Starter Culture for Plant-Based Fermented Beverages

Lactobacillus plantarum PMO 08 was evaluated as a starter culture for plant-based probiotic beverages. Its viability under various culture conditions and acidification ability in standardized tomato medium, fermentation parameters, and beverage properties were assessed. Lactobacillus plantarum PMO 08 could grow under various culture conditions; there was a high correlation between the incubation time to reach the optimal conditions and the inoculation concentration of lactic acid bacteria (LAB) (r2 = 0.997). Acidity (0.958 ± 0.002%) and LAB count (9.78 ± 0.14 Log10 CFU/mL) were significantly higher when fermented with L. plantarum than with the yogurt starter culture. A survival rate of 96% and 95% in artificial gastric juice and artificial intestinal juice, respectively, indicated that the probiotic requirements were met. The total polyphenol and glutamine content, and antioxidant activity increased after fermentation. The proline content significantly increased in L. plantarum PMO 08- fermented beverage. Thus, L. plantarum PMO 08 is an effective starter culture for non-dairy probiotic beverages whose functional quality may be improved by fermentation.

Enhanced Water Solubility and Oral Bioavailability of Paclitaxel Crystal Powders through an Innovative Antisolvent Precipitation Process: Antisolvent Crystallization Using Ionic Liquids as Solvent

Paclitaxel (PTX) is a poor water-soluble antineoplastic drug with significant antitumor activity. However, its low bioavailability is a major obstacle for its biomedical applications. Thus, this experiment is designed to prepare PTX crystal powders through an antisolvent precipitation process using 1-hexyl-3-methylimidazolium bromide (HMImBr) as solvent and water as an antisolvent. The factors influencing saturation solubility of PTX crystal powders in water in water were optimized using a single-factor design. The optimum conditions for the antisolvent precipitation process were as follows: 50 mg/mL concentration of the PTX solution, 25 °C temperature, and 1:7 solvent-to-antisolvent ratio. The PTX crystal powders were characterized via scanning electron microscopy, Fourier transform infrared spectroscopy, high-performance liquid chromatography–mass spectrometry, X-ray diffraction, differential scanning calorimetry, thermogravimetric analysis, Raman spectroscopy, solid-state nuclear magnetic resonance, and dissolution and oral bioavailability studies. Results showed that the chemical structure of PTX crystal powders were unchanged; however, precipitation of the crystalline structure changed. The dissolution test showed that the dissolution rate and solubility of PTX crystal powders were nearly 3.21-folds higher compared to raw PTX in water, and 1.27 times higher in artificial gastric juice. Meanwhile, the bioavailability of PTX crystal increased 10.88 times than raw PTX. These results suggested that PTX crystal powders might have potential value to become a new oral PTX formulation with high bioavailability.

Probiotic Properties and Antioxidant Activities of Pediococcus pentosaceus SC28 and Levilactobacillus brevis KU15151 in Fermented Black Gamju

Black gamju is Korean traditional beverage fermented with molds. The aim of this study was to assess the probiotic properties and antioxidant activities of novel Pediococcus pentosaceus SC28 and Levilactobacillus brevis KU15151 to develop black gamju with bioactive properties for health. Tolerance against artificial gastric juice and bile salts, adhesion ability on HT-29 cells of strains, and antibiotics susceptibility were evaluated as probiotics, and various enzyme productions were detected. The 2,2-diphenyl-1-picrylhydrazyl assay, 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonate, and β-carotene bleaching assay were used for antioxidant activity of samples. The tolerance of both strains to artificial gastric juice and bile salts (Oxgall) was more than 90%. Additionally, both strains did not produce β-glucuronidase and were resistant to gentamicin, kanamycin, streptomycin, and ciprofloxacin. After fermentation of black gamju with each strain, the number of viable lactic acid bacteria increased to 8.25–8.95 log colony forming unit/mL, but the pH value of fermented samples decreased more (to pH 3.33–3.41) than that of control (pH 4.37). L. brevis KU15151 showed higher adhesion activity to HT-29 cells and antioxidant effects than P. pentosaceus SC28 in three antioxidant assays.

Fibrous g-C3N4@Tio2 Nanocomposites-Based Dispersive Micro-Solid Phase Extraction for Chromium Speciation in Cow Milk by ICP-MS after Digestion Treatment with Artificial Gastric Juice

Background Chromium is an interesting element because its toxicity depends on its speciation. Thus, knowledge of Cr speciation in cow milk is essential to human health. Objective This study aims to achieve real bioaccessible species in cow milk, including Cr(III), Cr(VI), residual, digestible, and total Cr. Methods Samples were treated with artificial gastric juice, followed by dispersive micro-solid phase extraction (DMSPE) combined with ICP-MS for Cr speciation. Fibrous g-C3N4@TiO2 nanocomposites (FGCTNCs) were used as a novel adsorbent for DMSPE. Results The method detection limits were 110 pg/g (Cr(III)) and 260 pg/g (Cr(VI)) for milk powder (0.1 g), and 5.1 pg/g (Cr(III)) and 13 pg/g (Cr(VI)) for liquid cow milk (2 mL). The relative standard deviations (RSDs), obtained by analyzing the standard solutions containing 1.0 ng/mL of the analytes in sequence for nine times, were 4.3% and 5.1% for Cr(III) and Cr(VI), respectively. Linearity was observed over the range of 4 magnitude orders with correlation coefficients better than 0.9961. The enrichment factor of 100 was obtained. The majority of Cr in the samples was transferred into digestion solution. The content of Cr(III) is much higher than that of Cr(VI) in the digestion solution. Conclusions This method has the advantages of reduced solvent consumption, less adsorbent dosage, and high extraction efficiency. It may become a valuable strategy for elemental species in food samples. Highlights The samples were treated with artificial gastric juice to avoid the inter-conversion of species. FGCTNCs exhibit the merits of N-rich functional groups and selective adsorption for the analytes.