CC BY
25
the anatomical and pathological changes in weight, size, color and texture compared with control group animals.
Furthermore, biochemical result of blood of test animals was significantly differed from control groups for altered indexes: it was decreased in 1.3 times of creatinine and in 1.7 times of associated bilirubin (P <0.05). Also there was observed a significant decline of such biochemical indicators ofblood tested animals: in 1.5 times of AST, 1.4 times of ALT, 1.4 times in urea and in 1.4 times of glucose relative to control (P<0.05).
Moreover, for biochemical indexes it was found that in the treated by GM product group, was increased as total protein value in 1.1-fold, as well the free bilirubin indices risen in 1.5 times relation to the control (P <0.05).
The result of blood hematological indexes shown that treated by GM soybeans group was reducing of hemoglobin in 1.2 times: 63.16 ± 7.06 g/L and 75.66 ± 8.46 g/l for test and control groups simultaneously; P <0.001). The number of leukocytes also was decreasing in 2.6 times (1.95 ± 0.36 x 10 9/L and 5.05 ± 3.59 x 10 9/L for test and control groups simultaneously; P<0.001).
At the same time into blood of rats from the test group was observed: a) the emerged in red blood cells of corpuscles Jolly (1.9±1.3%) and b) toxic granularity ofleukocytes (1.9 ± 1.3%), which were not observed in control group ofrats. The emergence of the toxic elements (corpuscles Jolly in erythrocytes and leukocytes toxic granularity) indicated the pathological process in animal body, and reduction of antitoxic characteristics of liver may be taken into account as evidence of decreasing in the immune resistance of organism.
For both groups was practically not found any traces of formation and/or development of tumor. In internal organs (heart, lungs) was not detected any abnormalities compared with control rats at their look and size as.
Conclusion: Certainly, GM products have great advantages over the original one by plan-ting. However, in this study we fed animals by GM soy additive as 0.003g per rat during 30 days and noticed two significantly effects on both: reproduction functions and immune system of treated by soybeans animals. The study confirms once again the statement of investigators that for each a new obtained by genetically engineered plant product must be subjected to careful investigations, which carried out by independent research groups [5; 7].
References:
1. Aripov A. N., Fesenko L. M. Clinical biochemistry. Methods. Tashkent, published by «Abu-Ali' Sino». - 2000. - 271 p.
2. Jarmuchamedova T.Yu. Semushina S. G. Pachomova I. A. et al. International rules on work with laboratory animals before the clinical researches//Toxicological Bulletin. - 2011. - 4 (109): 2-9. (I n Russin).
3. Karkishenko N. N. Gracheva S. V. Guidelines for experimental animals and alternative models biomedical researches. Moscow, published by «Profil», - 2010. - 241 p.
4. McCann M. C., LiuK., TrujilloW. A., Dogbert R. C. Glyphosate-tolerant soybeans remain com-positionally equivalent to conventional soybeans (Glycinemax L) during three years of yield-tes-ting//Journal of Agriculture and Food Chemistry. - 2005. - 53: 5331-5.
5. Kim S. H., Kim H. M., Ye Y. M. et al. Evaluating the allergic risk of genetically modified soy-bean//Yenisei Medical Journal. - 2006. -47: 505-12.
6. Yum H. Y., Lee S. Y., Lee K. E. et al. Genetically modified and wild soybeans: an immunological comparison//Allergy Asthma Proc. -2005. - 26: 210-6.
7. Cantani A. Benefits and concerns associated with biotechnology-derived foods: can additional researcher duce chi drumhead thirsts//Eur Rev ofMed Pharm Sci. - 2006. - 10: 197-206.
DOI: http://dx.doi.org/10.20534/ESR-17-1.2-95-98
Oripov Firdavs Suratovich, Samarkand State Medical Institute Chief of the Department of Histology, Cytology and Embryology,
Associate Professor, E-mail: [email protected]
Morphology of neuroendocrineimmune system of jejunum in early postnatal ontogenesis
Abstract: The morphology of the nervous, diffuse endocrine and immune apparatus of jejunum of 20 day old rabbits were comprehensively studied by using neurohistological, general histological and ultramicroscopic methods and found their morpho-functional improvements because of transition to new quality nutrition at this age. Keywords: jejunum, local regulatory apparatus.
Introduction. Study ontogenesis of the jejunum [2; 3; 4; 5] and its regulatory structures [1; 6] attracts attention ofthe investigators in the last years. However the complex study of regulatory structures of the jejunum wall in the period of the early postnatal ontogenesis which have a certain importance in order to elucidation their integrative role and interaction in the period of morphofunctional adaptation to the new existing and nutrition status have not enough studied.
Aim of investigation. The complex study of morphology of the local regulatory structures of the rabbits' jejunum in the period of adaptation for the qualitatively new nutrition method.
Materials and study design. Morphology of the intramural nerve, diffuse endocrine apparatus and immune structures of jejunum of 16 rabbits of 20 days of age have been studied. Fixation of material in formalin and embedding material in paraffin has been
carried out by the general accepted method. Paraffin sections were colored by the use of hematoxylin-eosin method. Cryostat sections of the material fixed in 12% of neutral formalin have been processing by silver nitrate according to the Bolshovskiy-Gross and Campos methods. Cryostat sections from wandering material have been processing by the solution of glyoxylic acid in order to determine adrenergic nerve structures and fluorescent endocrine cells. It is needed to mention that in specimens processed by Grimelius method immune structures are clear and very well determined. Specimens processed by glyoxylic acid have been examined and photographed on digital camera through luminescent microscope LUMAM-IZ with the use FS.1-6 and FS.1-4 filters. The structures' density was determined on Standard Square in the vision field of microscope.
The results of the own investigations. The study ofmorphology ofthe intramural nerve apparatus ofjejunum ofrabbits of20 days ofage showed that in this period of postnatal development nerve elements of
jejunum have been characterized by well apparent typological differentiation ofnerve cells. Increasing amount offunctionally "matured" nerve cells (they used to have the well-developed neurofibrillary apparatus, axons are differentiated, tigroid substance has clumpy appearance and nucleus is localized in the center of the cell) has been determined in the ganglions of intramural nerve plexuses. Number of neuroblasts in ganglions is decreased; however in each intramural ganglion it can be determined the certain amount of them. Nerve cells of the first Dogel's type (neurocytes with long axons) shows high sensitivity to silver nitrate. Hyper-impregnated and comparatively thick axon ofthese cells is often entered to the composition ofnerve column and until certain distance is sharply distinguished from the left nerve fibers ofganglion by thick caliber and intensive coloration. Dendrites are usually ramified inside intramural ganglion. Intramural ganglions are clearly limited and localized at the same direction as nerve column which also well impregnated and determined in the comparison with previous ages (figure 1).
¿Jiá/ "-¿s
Figure 1. Intramural nerve ganglion of subcutaneous plexus of jejunum of rabbit of 20 days of age. Fascicles of nerve fibers (A) and nerve cells (B) of the different degrees of impregnation can be seen. Bolshovskiy-Gross painted by aluminous carmine. Ob. 20, ok. 10
Some ganglions contain a little amount of nerve cells, but it is found ganglions containing several ten nerve cells. Effectors of nerve endings on the body of nerve cells in the form of pericellular apparatus around nerve cells or separated "rings" localizing on the
body of the nerve cell of the first type, sometimes in the interneuronal cavity can be repeatedly determined at this age. Sometimes transitory synapses on the body of neurons of the first type can be observed (figure 2).
A
B
Figure 2. Nerve ending in the form of "ring" (A) and transitoty synapse ending (B) on the long-axon neurocytes of jejunum of rabbit of 20 days of age. Impregnation by Campos. 0b.40, ok.10
Adrenergic nerve fibers as well as the other nerve structures at this age are clearly formed and as if it is happened morphologic "leap" in the structure and differentiation of these nerve structures, they are accurate and fluorescence degree is high (figure 3). During the analyses of the more light sections and thin areas of the total specimens we have determined phenomenon which should be taken into consideration, i. e. in fact that adrenergic perivascular plexus is accompanied
branches of arteries before their bifurcations and it is absent in capillaries. Fluorescent endocrine cells of the jejunum of the rabbits at this age have been clearly determined in the epithelium of crypts and fringes. Distribution density of these endocrine cells in the j ejunum of the rabbits of 20 days of age was on average 7,2+0,34. They often have rounded or oval with elongated apical part form reminding fluid drop. Process of cells' extrusion of secretion could be seen in some cells.
/
//r
Wji
Figure 3. Perivascular adrenergic plexus around the arteries of different calibers of jejunum of rabbit of 20 days of age. Total specimen. Processing by glyoxylic acid. Ob. 20, gomal 3
Therefore, to 20 days of age postnatal ontogenesis intramural nervous apparatus of jejunum is morphofunctional differentiated. The components of the adrenergic nervous apparatus provided enough neurotransmitter. Argyrophil endocrine cells of the jeju-
num in rabbits of this age are located in the epithelium of the crypts and villi. Electron microscopic study of endocrine cells showed that in this period in their cytoplasm secretory granules is clearly visible, which often found on basal parts endocrinocytes (figure 4).
Figure 4. Localization of major polymorphic granules on basal pole APUD cell (arrow). Electron microphotograph. Inc. 6000
Immune wall structure of the jejunum of rabbits of 20 day of age is presented in separate clusters of lymphoid tissue, diffusely located lymphocytes and intraepithelial lymphocytes. Submucosal wall of rabbits' jejunum of this age have separate clusters of lymphoid tissue, a part of which contains more mature lymphocytes, in contrast, from those structures of rabbits jejunum of previous age.
The central part of the lymphoid aggregates is lighter and contains lightly colored, relatively large cells. The peripheral part is painted basophilic. This testified about the appearance of reactive reproduction center in these clusters. However, lymphoid clusters have no clear boundaries. The average diameter of lymphoid clusters of jejunum of rabbits of 20 day of age on averaged was 273,0±2,94 mcm.
A
B
Figure 5. Lymphoid nodule with emerging light center (A) and high density of interstices lymphocytes in the stroma of the villi (B) of the mucous membrane of the jejunum of rabbit of 20 days of age. Staining by hematoxilin-eosin. 0к.10, оb.20
At the base of the villi in the submucosal basis, also there is a high density arrangement of interepithelial lymphocytes, unlike from parts of submucosal basis of crypts (figure 5). In rabbits of 20 days old, sharp increase in the amount of diffusely located lymphocytes has been observed. They are located in the lamina propria and in the submucosal basis of the jejunum. Intratissular lymphocytes was significantly more in the stroma of villi (12,3±0,16.), in contrast to the lamina propria of the region of crypts (7,06 ± 0,12).
The base of most of the villi because of the high density of intratissular lymphocytes consisting mainly of a large number of mature lymphocytes has dark basophilic coloration. These changes seem to be associated with a change in the rabbits' diet and participation villi in the digestion process.
In the epithelium of the mucous membrane of the jejunum rabbits of 20 days of age contain more intraepithelial lymphocytes. They are mainly found in the epithelium of the villi and are lo-
cated both above and on the level of mucosal epithelial nuclei. Intraepithelial lymphocytes mainly consist of mature lymphocytes with large basophilic colored nucleus of rounded shape. These data also indicate the active involvement of the villi epithelium than that of crypts in the digestion process of the food of a new character which differs in composition from the food of rabbits of 10 days old.
Conclusion. Studying the local immune apparatus jejunum of rabbits of 20 days of age of early postnatal ontogenesis, we can conclude that the immune structure of the jejunum of this age differ from previous ages, both in quantitative and qualitative terms, and they are more formed. This is apparently due to the transition of the rabbits of this age to the qualifiedly changed nature of the food (mixed feeding begins at this age). Thus, all structures of the neu-roimmuneendocrine regulatory apparatus ofjejunum of20 day-old rabbits are morphofunctionally formed.
References:
1. Karavaev P. G., Toymentsev V. V. Neuroimmune relationship.//International journal of applied and fundamental investigations. -2012. - No 1. - P. 65.
2. Sverdlova A. V. Influence of prenatal ethanol chronic intoxication for the development of the rats' generation in the early periods of postnatal ontogenesis.//Fundamental investigations. - 2008. - No 1. - P. 148-150.
3. Smekalina O. S., Brukhin G. V. Morphofunctional characteristic of endocrine cells of mucous membrane of gastro-intestinal tract of generation of female rats with chronic experimental injures of the liver.//Bulletin of South-Ural State University. - 2010. - No 37. - P. 51-56.
4. Chumakov VYu., Pazuk P. V. Morphofunctional features of lymph flow of jejunum of dogs on the stages of postnatal ontogenesis.//Bul-letin of KrasSAU. - 2013. - No 6. - P. 105-107.
5. Yuldashev A.Yu., Rakhmatova N.Kh., Nishonov A. A., Nugmanova U. T., Yuldasheva Z. Z., Chinieva M. I. Space-timed organization of the proliferation processes and migration of lymphoid cells of lymph nodes of Peyer's gland of jejunum.//Journal of theoretical and clinical medicine. - 2015. - No 5. - P. 28-32.
6. Yaglov V. V., Yaglova N. V. New conception of biology of diffuse endocrine system: outcomes and perspectives of its study.//Bulletin of Russian academy of medical science. - 2012. - No 4. - P. 74-81.
DOI: http://dx.doi.org/10.20534/ESR-17-1.2-98-100
Oripov Firdavs, Samarkand State Medical Institute, PhD in medicine, department of histology, embryology and cytology E-mail: [email protected]
Age morphology of immune structures of rabbit jejunum in the period of the early postnatal ontogenesis
Abstract: Based on general histological and histochemical methods of investigation we have studied the comparative morphology of immune structures of jejunum in young rabbits in the different periods of early postnatal ontogenesis. The study has determined presence of morphological and morphometric differences between them, although the general principles of their structure has been identical. The localization density of intra-tissue lymphocytes has demonstrated intensive increase until 20 days of postnatal ontogenesis.
Keywords: Jejunum, immune structures, age morphology.
Introduction. The functioning of the organs of digestive system after birth of the animals led to the some adaptation-morphologic changes of the immune structures, although the general appropriateness and principles of structural organizations were still without changes. In the realization of die immune reaction it is important to know the development lymphoid tissue in ontogenesis (Zavarin, 1985). There have some scientific works dedicated to the study of the immune structures of jejunum in the normal condition
(Novakovskaya, Archakova, and Gurin, 2004; Chava, 2004, 2008) and during the influence some factors (Azizova, Hasanov, and Tu-lemetov, 2004; Tuhtaev, Azizova, and Hasanov, 2004; Chava, 2006; and others). Although in die structure of the immune structures there have numerous parallelisms, perhaps in each class of vertebrates complication of this organization could be reached indepen-dendy (Gorishina and Chaga, 1990). Jejunum is the important part of the digestive tube where the final chemical processing of chime
