Exacerbation Of Cytomegalovirus Infection Research Articles

ПРОЦЕССЫ ЛИПОПЕРОКСИДАЦИИ И СИСТЕМА АНТИОКСИДАНТНОЙ ЗАЩИТЫ В КРОВИ ПУПОВИНЫ НОВОРОЖДЕННЫХ ПРИ ЦИТОМЕГАЛОВИРУСНОЙ ИНФЕКЦИИ В ПЕРИОД БЕРЕМЕННОСТИ

The aim of the work is to study the activity of lipoperoxidation processes and antioxidant protection system in the umbilical cord blood of newborns from CMV-seropositive women with exacerbation of infection in the third trimester of pregnancy. Lipid peroxidation activity and antioxidant status in 30 newborns from CMV-seropositive women with exacerbation of infection in the third trimester of pregnancy and in 25 from CMV-seronegative women were studied. Erythrocytes were used to determine the products of oxidative modification of lipids; antioxidant status was evaluated in the blood plasma of the umbilical vein. The content of TBK-active products was determined by fluorometric method, lysophosphatidylcholine by thin-layer chromatography, arachidonic acid by gas-liquid chromatography. Antioxidant status was assessed by α-tocopherol content and superoxide dismutase activity. Registration of optical densities and fluorescence was carried out using a spectrofluorometer. The study found an increase in the content of red blood cells of the umbilical cord of newborns from CMV-seropositive women with an exacerbation of infection in the third trimester of pregnancy of TAC-active products in 2.1 times (p˂0.001), of lysophosphatidylcholine in 1.9 times (p˂0.001) and of arachidonic acid in 1.5 times (p˂0.001). The antioxidant status of umbilical cord blood was characterized by 1.5 times decrease in α-tocopherol (p<0.01) and 1.3 times decrease in superoxide dismutase activity (p˂0.001) compared with similar indices in the group of newborns from CMV-seronegative women. The results of the study show that the exacerbation of cytomegalovirus infection in the third trimester of pregnancy is accompanied by the development of oxidative stress and tension in the system of antioxidant protection in the blood of the umbilical cord of newborns due to increased consumption of α-tocopherol and suppression of superoxide dismutase activity.

ПРОЦЕССЫ ПЕРОКСИДАЦИИ ЛИПИДОВ В ЭНДОТЕЛИИ АРТЕРИИ ПУПОВИНЫ ПРИ ЦИТОМЕГАЛОВИРУСНОЙ ИНФЕКЦИИ

The purpose of the work is to study the processes of peroxidation of lipids and their influence on the structure of the endothelium of the umbilical artery of newborns from mothers who had experienced exacerbation of cytomegalovirus (CMV) infection in the third trimester of pregnancy. For the object of the study there were taken 45 umbilical cord samples during childbirth from women who had acute CMV infection in the third trimester of pregnancy (main group), and 25 umbilical cord samples from healthy women (control group). Clinical diagnosis of primary CMV infection was established by the presence of IgM antibodies to CMV and low-avid IgG in the peripheral blood (avidity index <50%), as well as CMV DNA detected by polymerase chain reaction in blood or urine; exacerbation of chronic CMV infection by the presence of IgM to CMV, high-avid IgG (avidity index >65%), and CMV DNA in scrapes taken from buccal epithelium. Lipid peroxides in the endothelium of the umbilical artery were detected by the Winkler-Schulze method. Morphological examination of the umbilical cord was performed by histochemical methods. The results of histochemical studies indicate that the exacerbation of CMV infection in the third trimester of pregnancy is associated with oxidative stress that causes the depletion of the protective systems and oxidant potential, which leads to the development of significant oxidative destruction of the endothelium of the umbilical cord artery of newborns. The revealed morphohistochemical changes in umbilical cord tissues in CMV infection can affect the functional state of this organ and be a morphological substrate for the appearance of its dysfunction, mainly the basis for the violation of fetoplacental blood circulation causing the development of hypoxia.

DISTURBANCE OF THE PROGESTERONE AND ITS METABOLITES SYNTHESIS IN THE DEVELOPMENT OF NEUROLOGICAL DISORDERS IN CHILDREN AFTER CYTOMEGALOVIRUS INFECTION DURING PREGNANCY

Cytomegalovirus (CMV) infection is one of the causes of congenital neurological disorders and the virus itself is the most common viral agent causing an imbalance in the production of placenta progesterone and its neuroactive metabolites – allopregnenolone and 5α-dihydroprogesterone. The aim was to evaluate the concentration of progesterone and its metabolites – 5α-dihydroprogesterone and allopregnenolone – in placenta during exacerbation of CMV infection in the first trimester of pregnancy, and the impact of these disturbances on the development of neurological disorders in children. We examined 30 pregnant women with exacerbation of CMV infection in the first trimester of pregnancy and 30 pregnant women with latent disease; and later their newborns. The enzyme immunoassay was used to determine concentration of progesterone in placenta; the histochemical method – to determine 5α-dihydroprogesterone and allopregnenolone. Newborns underwent neurosonography studies. Exacerbation of CMV infection in the first trimester of pregnancy decreased progesterone in placenta by 1.3 times, 5α-dihydroprogesterone – by 1.73 times and allopregnenolone – by 2 times. Ultrasound examination of the brain showed ventriculomegaly, periventricular ischemia, and pseudocysts in newborns up to one year from mothers with exacerbation of CMV during pregnancy. Later, minimal brain dysfunctions were manifested by motor disorders, increased general, vegetative excitability, and a tendency to digestive and sleep disorders. The data obtained indicate that the exacerbation of CMV infection in the first trimester of pregnancy is interrelated with a decrease in the concentration of progesterone and its metabolites (5α-dihydroprogesterone, allopregnenolone) in the placenta and development of neurological dysfunction in newborns.

FREE-RADICAL OXIDATION AND OXYGENATION OF HEMOGLOBIN AT EXACERBATION OF CYTOMEGALOVIRUS INFECTION IN THE SECOND TRIMESTER OF PREGNANCY

The aim of the study was to assess free radical processes and their effect on oxygenation of hemoglobin in the blood of 18-21 weeks pregnant women with exacerbation of cytomegalovirus infection. We examined 40 pregnant women with exacerbation of cytomegalovirus infection and various levels of specific antibodies (IgM) at 18-21 weeks and 30 pregnant women without cytomegalovirus. The spectrophotometric method was used to determine the content of oxyhemoglobin, methemoglobin, thiobarbituric acid (TBA)-active products, superoxide dismutase in blood erythrocytes; thin layer chromatography – phosphotidylcholine, lysophosphatidylcholine; gas-liquid chromatography – arachidonic acid. Exacerbation of cytomegalovirus infection changes the activity of free radical oxidation processes, the severity of which is determined by the level of IgM antibodies and is manifested by a decrease in superoxide dismutase (p = 0.000) and phosphatidylcholine (p = 0.000), an increase in phospholipase A2 (p = 0.000), lysophosphatidylcholine (p = 0.000), arachidonic acid (p = 0.000) and TBA-active products (p = 0.000). Accumulation of superoxide anion radical and products of lipoperoxidation in erythrocytes decreases oxyhemoglobin (p = 0.000) and increases methemoglobin (p = 0.000). Exacerbation of cytomegalovirus infection at 18-21 weeks is associated with the enhancement of free radical lipid oxidation and a deficiency in the antioxidant activity of superoxide dismutase. It decreases oxygen transport properties and increases phagocytosis by red blood monocytes; and pregnant women develop further hemic hypoxia.

ИЗМЕНЕНИЕ ПОКАЗАТЕЛЕЙ СВОБОДНО-РАДИКАЛЬНОГО СТАТУСА, АНТИОКСИДАНТНОЙ ЗАЩИТЫ И МОРФОЛОГИЧЕСКИЕ ИЗМЕНЕНИЯ ЭРИТРОЦИТОВ ПЕРИФЕРИЧЕСКОЙ КРОВИ БЕРЕМЕННЫХ ПЕРВОГО ТРИМЕСТРА С ЦИТОМЕГАЛОВИРУСНОЙ ИНФЕКЦИЕЙ

The aim is to study the features of changes of free-radical status, antioxidant protection and morphologic changes of erythrocytes of the peripheral blood of pregnant women of the first trimester with exacerbation of cytomegalovirus infection (CMVI). 35 pregnant women with exacerbation of CMVI in the first trimester of gestation (the main group) and 35 pregnant women without this pathology of the same age and term as the patients of the main group were examined. Erythrocytes of the venous blood stabilized by heparine as well as blood plasma became the material of the study. There has been studied the condition of lipid peroxidation (LP) by the contents of diene conjugates and active products reacting with thiobarbituric acid; the state of nonenzymatic antioxidant protection by the contents of α-tocopherol as well as of the enzyme of erythrocytes (superoxide dismutase); morphologic changes of erythrocytes. It was found out that the exacerbation of CMVI in the first trimester of pregnancy is associated with activation of LP processes which include modification of not only primary, reverse stages of this process – the increase of the concentration of diene conjugates (p<0.001), but they lead to an increased formation of secondary active products reacting with thiobarbituric acid (p<0.001), intensifying the destructive action on the membrane of erythrocytes. Primary and secondary products of LP against the decrease of concentration of erythrocyte superoxide dismutase (p<0.001) and α-tocopherol (p<0.001) are accumulated in increasing concentrations and become potential factors of erythrocytes damage causing their functional and metabolic inferiority. Metabolic disturbances and damages of cellular membranes lead to the increase of population of echinocytes, target-like and degenerative forms of erythrocytes, which can be accompanied by a high probability of intravascular hemolysis and contribute to the development of hemic hypoxia complicating the infectious process and the course of pregnancy. Within the present hyperoxidation and suppression of antioxidant protection that lead to the worsening of the rheology of blood, women with exacerbation of CMVI in the first trimester of pregnancy should be prescribed the medicine stabilizing the lipid exchange.

АРХИТЕКТОНИКА ВЕНОЗНОГО РУСЛА ПЛАЦЕНТЫ У ЖЕНЩИН, ПЕРЕНЕСШИХ ОБОСТРЕНИЕ ХРОНИЧЕСКОЙ ЦИТОМЕГАЛОВИРУСНОЙ ИНФЕКЦИИ НА ПЕРВОМ ТРИМЕСТРЕ БЕРЕМЕННОСТИ

The aim of the study was to assess the architectonics of the venous bed of the fetus part of placenta from women with the exacerbation of cytomegalovirus (CMV) infection in the first trimester of pregnancy. There were examined 75 patients after the term birth, among them there were 25 seropositive women with the exacerbation of CMV infection in the first trimester of pregnancy without clinical signs of threatened miscarriage; 23 seropositive women with the exacerbation of CMV infection in the first trimester of pregnancy and clinical signs of threatened miscarriage; 27 seronegative women without threatened miscarriage. The architectonics of the venous bed of the fetus part was studied with the pressing of contrast (red lead oxide in the linseed oil) through the umbilical vein into the vessels and placenta cotyledons. The roentgen shots were done at the device RUM-20 M “Sapphire”. During the research of the venous bed of the fetus part of placenta from women with the exacerbation of CMV infection and threatened miscarriage in the first trimester of pregnancy there was identified a velamentous attachment of the umbilical cord with the segmentary narrowing and venous varices as well as with the decrease of the outer diameter of veins of preconfluent zone of the fetus part of the organ. There is an increase in the number of asymmetrically located biconfluent venous vessels, the quantity of marginally located and blindly finishing curved vessels of veins and venous roots, which does not provide the effective blood flow and leads to the ischemia of the peripheral (edge) part of placenta. This reduces the delivery of oxygen and metabolites to the fetus, is accompanied with the development of plethora, stasis, thrombosis and inflammation in veins and arteries, as well as the increase of the risk of partial non-progressive separation of normally placed placenta and its partial intimate attachment to the uterus.

ВЛИЯНИЕ ЦИТОМЕГАЛОВИРУСНОЙ ИНФЕКЦИИ НА ПРЕОБРАЗОВАНИЕ ПРОГЕСТЕРОНА В МЕТАБОЛИТ 5α-ПРЕГНАН-3,20-ДИОН В ПЛАЦЕНТЕ РАННИХ СРОКОВ БЕРЕМЕННОСТИ

The aim was to study the activity of progesterone and 5α-pregnan-3.20-dione conversion in the placenta at the exacerbation of cytomegalovirus infection (CMVI) during the first trimester of pregnancy. The study material included 50 villous chorions taken during medical abortions in the period of 8-10 weeks from women with exacerbation of chronic CMVI during pregnancy. The control group had 35 villous chorions of pregnant women of the same gestation period with chronic CMVI in the latent stage. Diagnosis of CMVI was performed taking into account the activity of CMVI with the immune-enzyme method by the level of antibodies of IgM and antibody titers of IgG in the paired serums in dynamics in 10 days. The progesterone content was determined at the “Stat Fax-2100ˮ spectrophotometer. The activity of 4-pregnene-3.20-dione dehydrogenase and 5α-pregnane-3.20-dione dehydrogenase was assessed by the histochemical method. Quantitative evaluation of the reaction products was carried out on sections under the microscope “Meiji Technoˮ connected to the software-hardware complex “SCION Corporationˮ. It was found out that progesterone content in villous chorions of women with reactivation of CMVI during pregnancy was reduced to 21.5±2.73 nmol/L in comparison with the control group (53.4±3.70 nmol/L, p<0.001). The disturbance of progesterone inversion intensiveness in placenta was proved by the decrease of enzyme activity: of 4-pregnene-3.20-dione dehydrogenase till 15.9±0.47 standard units (in control it was 28.6±0.58 standard units, p<0.001) and of 5α-pregnan-3.20-dione till 16.2±1.11 standard units (in control it was 25.0±1.91 standard units, p<0.001). Reduction in the intensity of progesterone conversion into 5α-dihydroprogesterone (5α-pregnan-3.20-dione) in the placenta with the exacerbation of CMVI can negatively affect the formation and functioning of the fetal nervous system.

ОЦЕНКА ПОКАЗАТЕЛЕЙ ЛИПИДНОГО МЕТАБОЛИЗМА У РОЖЕНИЦ И ИХ НОВОРОЖДЕННЫХ ПРИ ЦИТОМЕГАЛОВИРУСНОЙ ИНФЕКЦИИ

In order to study the lipid metabolism of a mother–newborn system, there were examined 35 women in labor who had the exacerbation of chronic cytomegalovirus infection with the titer of IgG antibodies to cytomegalovirus 1:1600 in the III trimester of gestation and the same number of newborns (the main group). The control group consisted of 35 healthy pregnant women and their newborns. In maternal blood plasma and neonatal umbilical cord blood, total cholesterol content, triglyceride, lipoproteid cholesterol of high (HDL) and low (LDL) density were studied; in the membrane of erythrocytes the concentration of arachidonic acid and lysophosphatidylcholine was examined. It was found that in comparison with the control group at exacerbation of cytomegalovirus infection during gestation there was a significant transformation of lipid metabolism not only in the mother´s blood, but in the blood of the umbilical cord of newborn children, which was revealed through the reduction of the content of total cholesterol (in mother it was by 21%, in the newborn it was by 35%, p<0.001) and cholesterol of LDL (in mother by 24%, in the newborn by 36%, p<0.001); through an increase in the concentration of triglycerides (in mother by 28%, in newborn by 21%, p<0.001); cholesterol of HDL (in mother by 24%, in the newborn by 46%, p<0.001) and chaotropic fractions of phospholipid bilayer of membranes of red blood cells – lysophosphatidylcholine (in mother by 69%, in the newborn by 76%, p<0.001); arachidonic acid (in mother by 62%, in the newborn by 34%, p<0.001). The disturbance of the lipid composition contributes to pathogenesis of CMV infection and allows further conceptual approaches to prevention and treatment of viral infection during pregnancy. The study results suggest the feasibility and necessity of including drugs with antioxidant, antihypoxic action and the ability to regulate lipid metabolism in the complex therapy of pregnant women.

Energy Metabolism in the Placenta and the Role of Disturbances in the Development of Placental Insufficiency at an Exacerbation of Cytomegalovirus Infection

Determine the characteristics of placental energy metabolism and to establish its role in the development of placental insufficiency at an exacerbation of cytomegalovirus (CMV) infection in 25–28 weeks of gestation. In a prospective study of the case-control type included pregnant, delivery on term of 37–38 weeks. The sample of 50 pregnant women, including 25 CMV-seropositive with exacerbation of CMV infection at 25–28 weeks of gestation and with the titer of IgG antibodies to CMV 1: 1600 at the time of the study and 25 CMV-seronegative women the same pregnancy. The study was conducted at the obstetric department of pathology of pregnancy and laboratory «Etiopathogenesis mechanisms and recovery processes with non-specific lung diseases» Far Eastern Scientific Center of Physiology and Pathology of Respiration together with the urban maternity ward at City Hospital in the period from 2014 to 2015. The activity of pyruvate dehydrogenase, α-ketoglutarate dehydrogenase and a dehydrogenase lipoic acid was determined by histochemical methods on cryostat sections of fresh frozen tissue placenta by the method of R. Lilly. Evaluation of the intensity of histochemical reactions carried out by the program cytophotometry Scion. The morphology of the placenta was studied in paraffin sections stained with hematoxylin Böhmer-eosin, van Gieson’s picrofuchsin and alcian blue by Steedman. Exacerbation of CMV infection at 25–28 weeks of gestation leads to a decrease in the intensity of the histochemical reaction of pyruvate dehydrogenase in 2.4 times, lipoic acid dehydrogenase — in 2.9 times, and α-ketoglutarate dehydrogenase — in 1.5 times in the syncytiotrophoblast villous placenta. The placental morphological structure study showed villi in a state of death or necrotic changes, as well as increasing the number of avascular immature villi. In the maternal part of the placenta were marked constriction clearances, hypertrophy of muscle and connective tissue layers blood vessels. The findings suggest that the exacerbation of CMV infection at 25–28 weeks of pregnancy causes a decrease in the intensity of energy metabolism in the placenta by suppressing the activity of the enzymes α-ketoglutarate dehydrogenase and pyruvate dehydrogenase complex, which is accompanied by disturbances of the morphological structure of the placental barrier, the development of placental insufficiency.

Effect of Annexin V on Development of Apoptosis in Placental Syncytiotrophoblast Nuclei during Exacerbation of Cytomegalovirus Infection in the Third Trimester.

During exacerbation of cytomegalovirus infection during pregnancy, cytomegalovirus proteins penetrate across the external membrane of placental villous syncytiotrophoblast and suppress activities of ATPase and Bcl-2 protein. Simultaneous increase in the content of annexin V and caspase 3 leads to structural disturbances in DNA in syncytiotrophoblast nuclei and stimulates their apoptosis. Structural abnormalities in the syncytiotrophoblast during exacerbation of cytomegalovirus infection are dangerous for fetal development.

A Method for Detection of Cholesterol Synthesis in the Placenta during Exacerbation of Cytomegalovirus Infection.

Histochemical analysis showed that exacerbation of cytomegalovirus infection even at early terms of gestation (weeks 5-6) suppresses synthesis of mevalonic acid, squalene, and 7α-dehydrocholesterol dehydrogenase in the placenta. Inhibition of cholesterol biosynthesis in the placenta at the early stages of gestation decreases production of sex hormones, reduces cholesterol concentration in the maternal peripheral blood, and impairs cholesterol supply to the fetus, which is associated with the risk of placental insufficiency.

Activity of histidine in peripheral blood erythrocytes of pregnant women during exacerbation of cytomegalovirus infection.

We studied the effect of active cytomegalovirus infection on histidine content in peripheral blood erythrocytes of pregnant women at gestation weeks 20-22 and its involvement into hemoglobin oxygenation. Using the histochemical technique developed by us, we studied the distribution of products of specific reaction for histidine in peripheral blood erythrocytes of pregnant women. The percentage of histidine-positive erythrocytes and their area were evaluated. The relationship between the distribution of the products of the reaction for histidine in peripheral blood erythrocytes of pregnant women and the titer of anti-cytomegalovirus IgG was revealed. The histidine content in peripheral blood erythrocytes of pregnant women with active cytomegalovirus infection was reduced, which impaired heme binding to globin and decreased the formation of oxyhemoglobin.

Morphofunctional characteristics of the glutathione cycle in peripheral blood erythrocytes of pregnant women with a history of cytomegalovirus infection exacerbation during gestation.

We studied factors influencing peripheral blood erythrocytes of pregnant women with exacerbation of cytomegalovirus infection during gestation. Cytomegalovirus infection was associated with a decrease in the content of reduced glutathione in erythrocytes and glutathione peroxidase, which leads to H2O2 accumulation in the blood. Exacerbation of cytomegalovirus infection was associated with reduction of glutathione reductase and SOD activities. Erythrocyte antioxidant function and the peripheral blood discocyte count decreased. Accumulation of degenerative erythrocytes is fraught with the risk of anemia in pregnant women.