Side Effects of In ovo Given Sunset Yellow FCF on the Embryonic Development of the Spleen by Means of Histological and Enzyme Histochemical Methods

Background: E110 is one of the food colorants which is widely used in dairy products, fast foods, jam and dry beverage powders, aqueous drug solutions, tablets, capsules, toothpastes mouthwashes, hair care products and cosmetics. Doubts have accumulated in recent years that food additives might cause allergic reactions in humans or increase these ailments. In this study, side effects of Sunset yellow FCF (E110) on the embryonic development of chicken spleen were evaluated by means of histological, histomorphometrical and enzyme histochemical methods. Methods: In the study, 250 fertilized broiler eggs obtained from a commercial broodstock were used. Eggs were divided into 5 groups each having 50 eggs. Control eggs were either nontreated or distilled water injected via air sac. The eggs in the experimental groups were injected with 100ng/egg, 500ng/egg and 1.000ng/egg E110 prior to incubation. Blood and spleen samples were taken from randomly selected 10 eggs of each group at 11th, 15th, 18th and 21st days of incubation. Leukocyte formula, alpha-naphtyl acetate esterase-positive and acid phosphatase-positive lymphocyte percentages were determined in the blood samples and embriyonic development of the spleen was assessed in the tissue sections. Results: In the 500ng/egg and 1.000ng/egg E110 injected experimental groups, embryonic development of the spleen retarded, alpha-naphtyl acetate esterase-positive and acid phosphatase-positive lymphocyte rates were significantly depressed. Conclusions: Significant disturbances in the immune system functions of the affected animals might occure at post-natal period of their life.

PATHOHISTOLOGICAL PECULIARITIES OF THE FORMATION OF IRON OVERLOAD SYNDROME IN PATIENTS WITH STEATOHEPATITIS DEPENDING ON ITS ETIOLOGY

The aim – to study some histopathological features of the iron overload syndromeformation in patients suffering from steatohepatitis depending on its etiology.Material and methods. Liver biopsies were analyzed in 30 patients with nonalcoholicsteatohepatitis (NASH) and 20 patients with alcoholic steatohepatitis (ASH). Themorphological examination of the liver was carried out according to the standardmethods. The histopathological features of the liver were established on the basis ofthe method of staining with hematoxylin and eosin, with the confirmation, in case ofnecessity, by the histochemical methods - staining for fat (the method with Sudan-III) andcollagen fibers (the method with chromotropic-water blue according to N.Z Slinchenko).Microspectrophotometric studies were conducted using a cytological analyzer withsoftware "VideoTest - Size 5.0" (2000).Results. The same regularities for both trivalent iron and twovalent iron were revealed asa whole in the liver cells at histological investigations, that is in ASH the optical densityis at the average 1.3 (p<0.05) times higher than in NASH. The staining on trivalent ironis less intensive, than on the twovalent iron. There is probable iron overload syndrome inthe alcoholic steatohepatitis patients with increased uptake of it both 2+- and 3+- valentforms in the liver cells – reticuloendotheliocytes and hepatocytesConclusions. Iron overload syndrome with increased uptake of it for both 2+- and3+-valent takes place in the liver cells – endotheliocytes and hepatocytes – in patientswith alcoholic steatohepatitis. At alcoholic steatohepatitis the optical density withhistochemical staining on iron 2+ in reticuloendotheliocytes of the liver is 1.2 times andin hepatocytes – 1.3 times higher than at non-alcoholic steatohepatitis irrespective ofthe presence or absence of anemia. At alcoholic steatohepatitis the optical density ofhistochemical iron staining 3+ in reticuloendotheliocytes of the liver and hepatocytesis 1.3 times higher than in non-alcoholic steatohepatitis, and is irrespective of anemiapresence as well.

Autophagy and Apoptosis in the Midgut Epithelium of Millipedes

The process of autophagy has been detected in the midgut epithelium of four millipede species: Julus scandinavius, Polyxenus lagurus, Archispirostreptus gigas, and Telodeinopus aoutii. It has been examined using transmission electron microscopy (TEM), which enabled differentiation of cells in the midgut epithelium, and some histochemical methods (light microscope and fluorescence microscope). While autophagy appeared in the cytoplasm of digestive, secretory, and regenerative cells in J. scandinavius and A. gigas, in the two other species, T. aoutii and P. lagurus, it was only detected in the digestive cells. Both types of macroautophagy, the selective and nonselective processes, are described using TEM. Phagophore formation appeared as the first step of autophagy. After its blind ends fusion, the autophagosomes were formed. The autophagosomes fused with lysosomes and were transformed into autolysosomes. As the final step of autophagy, the residual bodies were detected. Autophagic structures can be removed from the midgut epithelium via, e.g., atypical exocytosis. Additionally, in P. lagurus and J. scandinavius, it was observed as the neutralization of pathogens such as Rickettsia-like microorganisms. Autophagy and apoptosis ca be analyzed using TEM, while specific histochemical methods may confirm it.

MORPHOMETRIC AND HISTOCHEMICAL RESEARCH OF MELISSA OFFICINALIS L. HERB FROM THE COLLECTION OF NIKITSKY BOTANIC GARDEN

The article presents the results of morphological-anatomical, morphometric and histochemical studies of the aerial part of Melissa officinalis L., a selection variety sample from the collection of Nikitsky Botanical Garden – National Science Center (NBG-NSC) of the Russian Academy of Sciences, Republic of Crimea. The data on the morphology, histochemistry and morphometry of the main diagnostic microscopic signs of the Melissa officinalis L. herb, which are crucial for establishing the authenticity of medicinal plant materials, and also show the localization of secretion of metabolites and the heterogeneous nature of Melissa essential oil, have been obtained.The aim. Is to determine the microdiagnostic signs and biometric characteristics of the Melissa officinalis L. variety sample, introduced in Nikitsky Botanical Garden, to study the possibility of using histochemical reactions for the analysis of tissues and secretory structures of the studied species.Materials and methods. The plant material was provided by Nikitsky Botanical Garden. The study of microscopic signs, morphometric characteristics as well as histochemical tests, were performed according to the methods of the State Pharmacopoeia of the Russian Federation, XIII edition, using a Micromed-1 light microscope, a Micromed Micrometer 3 LUM fluorescent microscope and a Digital Eyepiece microscope camera MD 3.2 digital camera. The photos were edited in the program Paint. NET.3.5.11.Results. The main diagnostic microscopic signs of the herb of the Melissa officinalis L. selection variety sample can be called as follows: simple unicellular conical, nonbranching hairs, simple multicellular conical hairs, capitate hairs with a multicellular leg and a funnel-shaped head and with a unicellular leg and a spherical head, essential oil glands of a radial-type with 6-8 excretory cells. These signs have been diagnosed in the crushed raw materials. A quantitative assessment of the morphological and anatomical signs of the leaf (leaf thickness, height of the cells of the upper and lower epidermis, stoma density per 1 mm2 of the epidermis), stem, calyx and corolla was carried out. The size of cells, stoma, trichomes, as well as the frequency of the location of trichomes have been analyzed, too. The presence and localization of lipids, polysaccharides, phenolic compounds, flavonoids, essential oil, terpenoids, alkaloids in tissues and trichomes of the herb have been studied by histochemical methods using light and fluorescent microscopy.Conclusion. Morphological and anatomical study of the herb of the Melissa officinalis L. selection variety sample from the Nikitsky Botanical Garden collection has been carried out, diagnostically significant microscopic signs of raw materials have been revealed and their biometric characteristics have been established. For the first time the localization of characteristic secreted biologically active substances in the tissues and secretory structures of the herb has been studied by histochemical methods. The results of the experimental studies complement the scientific literature data and can be used to confirm the authenticity, identification and standardization of the aerial parts of this variety sample.

MICROMORPHOLOGICAL ANALYSIS OF THE AERIAN ORGANES OF MARRUBIUM ALYSSON L. FROM FLORA OF TUNISIA

The micromorphological investigations ofMarrubium alysson L.growing on the territory of the Tunisian Republic have been conducted.The aimof the study was to conduct comparative ecological and morphological researches ofMarrubium alyssonaerian parts, to study an anatomical structure of caulifoliary system elements including stems, petioles, leaf margins a structure of the epidermis.Materials and methods.As an object of the research, dried-up herb and herbarium specimens ofMarrubium alysson L., growing on the territory of the Tunisian Republic, were used. The micromorphological investigations were conducted by morphological and histochemical methods.Results and discussion.Comparative ecological and morphological researches have been conducted. The anatomic structure of the elements of the caulifoliary system including stems, petioles, leaf margins has been studied. The anatomic structure of the epidermis has been studied, too. The diacytic type of the stomatal apparatus, numerous branched trichomes with multicellular basis, located both on top and lower leaf parts have been found out. The conducted researches are a fragment of complex researches of perspective raw plants of Tunisian flora.Conclusion. Comparative eсological and micromorphological researches have been conducted, the anatomic structure of the stems, petioles, leaf margins, and also the epidermis structure of top and lower leaf parts has been also carried out. The diacytic type of the stomatal apparatus, trichomes with multicellular basis located both on top and lower leaf parts have been found out. The collenchyme is located in the subepidermal zone of a stem, petioles and in the zone of the central vein of a limb. The revealed signs can be included into product specification files on herbal raw materials

Needle muscle biopsy: technique validation and histological and histochemical methods for evaluating canine skeletal muscles

This study evaluated the needle muscle biopsy technique using a 6G Bergström percutaneous needle combined with histological and histochemical methods to analyze the skeletal muscle of dogs. There are few studies about canine skeletal muscles and a lack of reports in the literature about tissue collection and analysis for canine species. Evaluation of 32 German Shepherd samples collected from the gluteus medius, at a depth of 3 cm, was performed. The choice of gluteus medius and the 3-cm depth provided good quantity fragments with sufficient sizes (3–5 mm), which permitted optimal visualization of muscle fibers. Myosin ATPase, at pH 9.4, 4.6, and 4.3, and SDH reactions revealed that all muscle samples analyzed had fibers in the classic mosaic arrangement, enabling counting and typification. The mean percentages of fibers were 29.95% for type I and 70.05% for type II. On the basis of these results, we concluded that the percutaneous needle biopsy technique for canine skeletal muscles is a safe and easy procedure that obtains fragments of proper sizes, thereby enabling the study of muscle fibers. Standardization of the muscle of choice and the depth of muscle sample collection significantly contributed to this success. This is an important method to evaluate muscle fiber types of dogs and diagnose important diseases affecting the skeletal muscles.