About "neutrophilic twins"

The term "neutrophilic twins" ("neutrophileZwillinge") V. Schilling designates neutrophilic leukocytes, which have a double size against the norm and two separate nuclei, similar in structure and shape. Castillo (Vireh. Arch. F. Path. An., Bd. 247, H. 1, 1923) found them, almost as a rule, with significant neutrophilic hyperleukocytosis, especially often in infectious diseases, in particular, pneumonia and scarlet fever. They are often also found in processes when an increased neoplasm of neutrophils occurs, as, for example, with myeloid leukemia, pernicious anemia, ulcerative-chronic endocarditis, etc.

PRODUCTIVE LONGEVITY AND REPRODUCTIVE QUALITY OF HOLSTEIN COWS OF FOREIGN BREEDING

The aim of this work is to determine the productive longevity of Holstein cows of different breedings. The results of the research showed that Finnish animals were superior in life expectancy and productive longevity, exceeding German breeding cows by 74 days (P<0.05) and Hungarian cows by 24 days, and by 0.06 and 0.12 lactations in the productive period respectively. At the same time, according to the level of average productivity per lactation and lifelong milk yield, the highest rates were established for the group of Hungarian cows. So, their average milk yield per lactation was higher: compared to other groups by 529.2 kg and 561.8 kg or 7.6 % and 8.1%; lifelong milk yield by 1305.5 kg and 996 kg (5.6 % and 4.2%); per one day of calving interval - by 0.64 kg and 0.8 kg (3.6 % and 4.6%) and per one day of life - by 0.56 kg and 0.60 kg or by 3.2 % and 5.6%. In the context of the group of cows of the German and Finnish breeding, no significant differences were noted by these indicators. The duration of pregnancy in experimental cows ranged from 285-289 days. The analysis of the obtained data indicates that the cellular protection indicators of Holstein cows are within the physiological norm. So, phagocytic activity, which expresses the percentage of active leukocytes participating in phagocytosis to the total number of counted neutrophilic leukocytes, is higher in animals of the German breeding compared with Hungarian animals - by 13.2% and with Finnish ones - by 13.4%. The phagocytic index is defined by the average number of phagocytosed microorganisms per active leukocyte and characterizes the intensity of phago-cytosis. In our experiment, the intensity of phagocytosis was higher in the German cows and amounted to 8.42, which exceeds this indicator by 11.4% compared with animals of the Hungarian breeding and by 3.3% compared to the Finnish animals.

NONSPECIFIC FACTORS OF ORGANISM RESISTANCE IN PATIENTS WITH ERYSIPELAS

Summary. Aim. The article is aimed to assess the nonspecific factors of the body’s resistance in patients with erysipelas and in the development of sepsis in these patients. Materials and methods. 114 case histories of patients who underwent inpatient treatment in the center of purulent-septic surgery in the Non-profit municipal enterprise “City Hospital № 3” in Zaporizhzhia for the period 2019-2020 were analyzed. According to the forms of the disease, the patients were distributed as follows: erythematous — 24 (21.0 %), bullous — 28 (24.6 %), phlegmonous — 48 (42.1 %), necrotic — 14 (12.3 %). Recurrent cases of the disease were noted in 21 patients. From them: with erythematous form — 4 (16.7 %), bullous — 5 (17.9 %), phlegmonous — 9 (18.7 %), necrotic — 3 (21.4 %). Among 10 patients, the disease was complicated by sepsis. 6 patients died, mortality was 60 %. Results and their discussion. Depending on the detected disorders, the patients were divided into three groups: with uniformly activated immune status; with a suppressive type of immune response; with a mixed type of immune status, where with a normal or suppressive cellular link of immunity, activation of some indicators of the humoral link against the background of a normal or reduced level of complement is determined, which indicates the sensitization of T-cell populations with an antigen and the development of autoimmune processes. Conclusions. An increase in complement levels is an indicator of active antigen-antibody binding reactions in patients with erysipelas. Weak NBT-reaction in patients with sepsis indicates depletion of the enzymological activity of neutrophilic leukocytes and, to some extent, can serve as a predictor of a lethal outcome. An increase in the rates of phagocytosis is a prognostically favorable sign indicating the effective removal of the antigenic material of immune complexes from the patient’s body.

MORPHOLOGICAL FEATURES OF CELLULAR INFILTRATION IN THE MUCOSA OF LARGE INTESTINE IN ULCERATIVE COLITIS AND IRRITABLE BOWEL SYNDROME

The aim is to improve morphological diagnostics, including differential, of UC and IBS, identifying morphological features of cellular infiltration in the mucous membrane ofthe large intestine. Material and methods: Autopsy and biopsy material – fragments of the mucous membrane of the large intestine was used in this study. All the material was divided into 5 groups. Group 1 included autopsy material from 6 cases, in which no general pathological processes in the gastrointestinal tract were detected during autopsy and microscopic examination. Group 2 included biopsy material from 34 patients with diagnosed UC of the 1st activity degree. Group 3 included the biopsy material of 27 patients with UC of the 2nd degree of activity. Group 4 included biopsy material from 19 patients, diagnosed with UC of the 3rd degree of activity. Group 5 included biopsy material from 82 patients with clinically diagnosed IBS. Histological, morphometrical, immunohistochemical and statistical methods of investigation were used. Results: The mucous membrane of the large intestine in patients with ulcerative colitis of varying degrees of activity, compared with the physiological norm, has pronounced infiltration by plasma cells, T-lymphocytes, B-lymphocytes, macrophages, mast cells, eosinophilic and neutrophilic leukocytes in the superficial parts of the epithelium, crypts,lamina propria. There is also an increase in the number and size of lymphoid follicles in the lamina propria. Predominant cellular elements in the infiltrate are plasma cells, T-lymphocytes, eosinophilic and neutrophilic leukocytes. The growth of ulcerative colitis activity leads to an increase the inflammatory cell infiltration in the mucous membrane of the colon, as evidenced an increase the density of cellular infiltrate; the severity of inflammatory changes in crypts and an increase in the number of crypt abscesses; a decrease the number of cases with focal infiltration in the lamina propria and an increase the number of cases with diffuse infiltration; the spread of inflammatory cell infiltration from the superficial parts of the lamina propria to its deep parts with the subsequent involvement of its entire thickness; an increase the central trends of the indexes of the severity of all cellular infiltration, infiltration by plasma cells, T-lymphocytes, macrophages, neutrophilic leukocytes. The mucous membrane of the large intestine in patients with irritable bowel syndrome has moderately pronounced cellular infiltration in the superficial epithelium andlamina propria, in comparison with the physiological norm. The number and size of lymphoid follicles increase. Inflammatory cell infiltration often spreads to the upper onethird or two thirds of the thickness of the lamina propria, characterized by the presence of plasma cells, T-lymphocytes, B-lymphocytes, macrophages, mast cells, eosinophilic and neutrophilic leukocytes. In this case, plasma cells, T-lymphocytes, mast cells and macrophages dominate. The indexes of the severity of all cellular infiltration, as well as infiltration by plasma cells, T-lymphocytes, B-lymphocytes, macrophages, mast cells, eosinophilic and neutrophilic leukocytes, increases in the mucous membrane of the large intestine in irritable bowel syndrome in comparison with the norm. In the mucous membrane of the large intestine in irritable bowel syndrome compared with ulcerative colitis of varying degrees of activity inflammatory cell infiltration is less pronounced. It often extends to one third or two thirds of the thickness of the lamina propria. There are fewer lymphoid follicles, cryptitis and crypt abscesses are not determined. The indexes of the severity of all cellular infiltration, as well as infiltration by plasma cells, T-lymphocytes, eosinophilic and neutrophilic leukocytes are lower. Conclusions: The revealed features of cellular infiltration in the mucous membrane of the large intestine make it possible to improve morphological diagnostics, including differential, of ulcerative colitis of varying degrees of activity and irritable bowel syndrome.

AB0135 THE MITOCHONDRIAL-RETICULAR NETWORK (MRN) OF NEUTROPHILIC LEUKOCYTES OF SYNOVIAL FLUID (SF) OF PATIENTS WITH SLE AND RA

Background:Objectives:It has been established that in cells, in particular in neutrophilic leukocytes of SF, mitochondria form a mitochondrial-reticular dynamic spatial network (MRN). MRN is the epicenter of apoptosis, reflecting structural and functional changes in the immuno-complex pathology in SLE and RA.Methods:SF was analyzed in patients: 10 SLE (43 ± 2.3 years), 13 RA (45 ± 1.6 years) and 8 donors (42 ± 3.7 years, postmortem). Neutrophilic leukocytes from the SF were isolated by standard methods and resuspended in a composition medium: 70 mM NaCl; 140 mM sucrose; 5.6 mM KCl; 10 mM pyruvate; 8 mM MOPS; pH = 7.4. The cell suspension was centrifuged for 5 min at 800 g. MRN was isolated by centrifuging the resulting supernatant for 15 min at 12 000 g. The resulting MRN fragments were resuspended in citrate-phosphate buffer (pH = 7.4) and used in experiments. The activity of adenosine monophosphate-activated protein kinase (AMPK) was evaluated by Western blotting. Quantitative determination of cytochrome C (Cyt C) was carried out by enzyme immunoassay method using the Human Cytochrome c Platinum ELISA kit (eBioscience, USA). Active forms of oxygen free radicals (AFRF) were registered by EPR. The swelling rate of MRN fragments was determined spectrophotometrically at 540 nm. The electrophoretic mobility (EM) of MRN fragments was determined by the automatic microscope “Parmoquant-2”.Results:MRN of neutrophilic leukocytes of the SF undergoes significant adaptive rearrangements during the development of SLE and RA (tab.1). On average, the expression of biochemical indicators of autophagy (AMPK), apoptosis (Сyt. C), necrosis (level of oxygen free radicals, low-amplitude swelling rate) increases by 2-3 times compared with the conventional norm. Particular attention should be paid to pathological changes in the electrokinetic potential of MRN, which determines the functional state of the SF as a whole as a colloidal system. Obviously, in SLE and RA, depletion of the energy of MRN (a sharp increase in the activity of AMRK), activation of free radical processes, disruption of intracellular ion homeostasis due to an increase in the rate of swelling of MRN as a manifestation of a compensatory-adaptive reaction. It ultimately leads to a decrease in electrokinetic properties of MRN. Thus EM is an integral indicator of physico-chemical properties and architectonics of MRN pointihg to the development of autoimmune pathology.Table 1.EXPRESSION OF INDUCTORS OF AUTOPHAGY, APOPTOSIS, NECROSIS AND ELECTROPHORETIC MOBILITY OF MRN FRAGMENTS OF NEUTROPHILIC LEUKOCYTES OF SF IN SLE AND RAExperience TermsAMPK,cond.unit/mg proteinCyt C,ng/mlAFRF, unit/mg proteinSwelling rate of MRN,/min ·mg proteinEM,m/v · secDonor (8)0,51±0,0523,7±5,47,3±2,40,177± 0,0041,58± 0,07SLE (10)1,73±0,04**49,3±6,5*21,3±5,1**0,435±0,005***0,35±0,05***RA (13)1,25±0,07**47,8±4.8*15,7±4,3*0,410±0,007***0,41±0,07***Notes: differences with the control norm: * - p <0.05; ** - p <0.01; *** - p <0.001.Conclusion:Endoplasmic stress occurs in SF cells during the development of SLE and RA, blocking of autophagy and apoptosis leads to a breakdown of neutrophilic leukocyte MRN, accumulation of high molecular products of tissue decay - phlogogens in the intercellular space, among which the expression in the context is characterized by proteins - chaperones Hsp 60-100. These processes are accompanied by a shift in the bioelectric homeostasis of MRN neutrophilic leukocytes, an increase in their swelling rate and a significant decrease in their electrokinetic potential. The described MRN reactions of neutrophilic leukocytes of the SF should be taken into account when developing pharmacologically induced apoptosis as a new approach in the treatment of autoimmune diseasesReferences:[1]Shishkin V. I. et al. Ann Rheum Dis 2017; 76: No 6, p.1077-1078; DOI:10.1136/annrheumdis-2017-eular5364Disclosure of Interests:None declared

Reactions of Subcutaneous Cellular Tissue in Response to the Implantation of a Polypropylene Endoprosthesis Modified with a Polycaprolactone Solution by Electrospinning

Objective: to study the morphological features of subcutaneous tissue in response to the implantation of a polypropylene mesh endoprosthesis (PSE) modified with a polycaprolactone solution by the method of electrospinning. Material and methods. The object of the study was modified polypropylene mesh endoprostheses implanted in experimental animals. The animals were removed from the experiment on the 3rd, 7th, 60th day of the experiment. Subcutaneous fatty tissue along with the implanted material and regional lymph nodes were fixed in a 10 % solution of neutral formalin, followed by histological posting according to the standard technique and filled with paraffin. Histological sections 3-4 μm thick were stained with hematoxylin and eosin. The morphological assessment of reactions to the implantation of the materials was carried out by means of the calculation of the cellular elements of inflammatory and connective tissue origin along the periphery of the implants (neutrophilic leukocytes, phagocytic neutrophilic leukocytes, degenerating neutrophilic leukocytes, eosinophilic leukocytes, lymphocytes, plasmacytes, histiocytes, macrophages, fibrocytes, fibroblasts, multinucleated giant cells).Results. The study of the cellular composition has showed a statistically significant increase in the content of fibrocytes (p = 0.002) and fibroblasts (p < 0.001) on the 7th day of observation around the modified PSE. On the 60th day, the number of neutrophilic leukocytes at the periphery of the modified PSE was lower than that of the control sample (p = 0.004). The enhancement of the fibroplastic properties of the samples modified with polycaprolactone by electrospinning was characterized by a less pronounced inflammatory reaction of the peripheral tissues.Conclusion. The obtained data are indicative of the fact that the modification of PSE with polycaprolactone reduces the reactive properties of endoprostheses and make it possible to recommend to introduce this method of PSE modification in clinical practice.