Effect of interlok, remantadine and biotop on normal monocyte functional activity and in acute viral hepatitis

The influence of interlok, biotop and remantadine on the functional activity of monocytes was determined with the use of the NBT test permitting the quantitative evaluation of this characteristic. In this study interlok was found to give promising results in the treatment of acute viral hepatitis, due both to its antiviral action and to the stimulation of the functional activity of monocytes. biotop proved to render a sharply pronounced stimulating effect on the functional activity of monocytes, thus enhancing antiviral resistance. Remantadine suppressed the functional activity of monocytes and increased their lesion by the virus, thus creating favorable conditions for virus persistence in monocytes.     

Effect of tactivin in vitro on lymphocytes from patients with acute hepatitis B

In vitro experiments have revealed that Tactivin increases the number of lymphocytes and their functional activity in patients with acute virus hepatitis B. Under the action of Tactivin the increase of theophylline-resistant and theophylline-sensitive T-lymphocytes occurs in patients with acute virus hepatitis B in a moderate or severe form. In a mild form of acute virus hepatitis B and in healthy persons Tactivin produces no in vitro changes in T-lymphocyte subpopulations.     

Identification and properties of complexes formed by myeloperoxidase with lipoproteins and ceruloplasmin

The first evidence of multi-component complexes formed by myeloperoxidase (MPO), ceruloplasmin (CP), and very low/low density lipoproteins (VLDL/LDL) obtained by electrophoresis, gel filtration, and photon-correlation spectroscopy (PCS) is presented in this paper. Complexes were observed when isolated MPO, CP, and VLDL/LDL were mixed and/or when MPO was added to the blood plasma. Complex LDL–MPO–CP was detected in 44 of 100 plasma samples taken from patients with atherosclerosis, and 33 of 44 samples also contained the VLDL–MPO–CP complex. MPO concentration in these patients’ plasma exceeded 800 ng/ml. Interaction of MPO with high density lipoproteins (HDL) was not revealed, as well as binding of CP to lipoproteins in the absence of MPO. Adding antibodies against apoB-100 to VLDL–MPO–CP and LDL–MPO–CP complexes results in release of lipoproteins. Using PCS the diameters of complexes under study were evaluated. By comparing concentrations of the components in complexes formed by MPO, CP, and lipoproteins their stoichiometry was assessed as 2VLDL:1MPO:2CP and 1LDL:1MPO:2CP. Lipoproteins affected the inhibition of MPO peroxidase activity by CP. The affinity of lipoproteins to MPO–CP complex was assessed using apparent dissociation constants determined as ～0.3 nM for VLDL and ～0.14 nM for LDL.     

The comparative characteristics of the indices of lymphocyte and neutrophil functional activity in patients with HIV infection and chronic viral hepatitis B]

In 34 patients with human immunodeficiency virus (HIV) infection at the asymptomatic stage and 29 patients with chronic viral hepatitis B at the period of exacerbation (of these 14 patients had chronic persistent hepatitis and 15 patients had chronic active hepatitis) the complex study of the functional activity of lymphocytes and neutrophils was carried out by cytochemical methods with the simultaneous determination of the content of immunoregulating lymphocyte subpopulations. In patients with chronic active hepatitis a decrease in the percentage and the absolute number of helper T-lymphocytes and the ratio of CD4/8 in comparison with those in patients with HIV infection were revealed. At the same time patients with HIV infection exhibited more pronounced decrease in the activity of all lymphocytic enzymes under study (neutrophil esterase, acidic phosphatase and succinate dehydrogenase in lymphocytes), as well as in the activity of myeloperoxidase and the content of cation proteins and glycogen in neutrophils in comparison with patients having chronic active hepatitis.     

The Effect of Magnetite Nanoparticles and Bacteria on the Activity of NADPH-Oxidase and Myeloperoxidase in Neutrophils of Human Blood

Differences in the oxygen-dependent reactions of neutrophil granulocytes (NGs) depending on the nature of the agent affecting the cells were revealed. In vitro magnetite nanoparticles (MNPs) cause suppression of the NADPH–oxidase activity of NGs, which manifests itself in falling rates of reactions (NBT test) both with the effect of MNPs on NGs alone and a combined effect (MNPs and zymosan), as well as in the reduction of the index of activation (IA) and functional reserve of neutrophils (FRN). However, the introduction of MNPs dose-dependently stimulates the activity of myeloperoxidase (MPO). Gram-positive (S. aureus 2879 M) and gram-negative (E. coli 321) bacteria caused a respiratory burst of neutrophils, which manifested itself in a significant increase in the number of NBT-positive cells in single and combined influences (bacteria and zymosan). The lack of differences in the reaction of cells on opsonized and nonopsonized bacteria and the decrease in IA and FRN suggest that NGs are at the maximum level of functionality. Both strains of bacteria caused activation of the MPO.     

The role of neutrophil myeloperoxidase in the development of inflammation induced by thermal skin burns

Luminol-dependent chemiluminescence (CL) of blood neutrophils stimulated with phorbol-12- myristate-13-acetate (PMA) and myeloperoxidase (MPO) activity of neutrophils and plasma have been investigated in children (n = 16) during the early period (1?7 days) after thermal skin burns exceeding 20% of total body surface. The CL level of stimulated neutrophils was higher in burn patients than in healthy children of the reference group (p lt; 0.01). Increased neutrophil MPO activity was found in 40% of patients, while increased plasma MPO activity was detected in 57% of patients. The albumin fraction isolated from plasma of burn patients increased the PMA-stimulated CL response of blood from healthy donors. These results suggest that the acute inflammatory response to the thermal burn causes neutrophil activation and MPO release into plasma. MPO-mediated modification of plasma proteins, particularly albumin, may stimulate neutrophil activation and provoke further inflammatory response of the body to the thermal injury.     

Characteristics of monocyte functional activity in acute viral hepatitis

The functional activity of monocytes in acute viral hepatitis has been studied. As revealed by the NBT test, circulating monocytes are damaged by viruses and/or virus-containing complexes. Proceeding from this fact, the percentage of monocytes containing virus inclusions should be considered as the percentage of monocytes damaged by the virus, and the coefficient of the inclusion of viral particles should be considered as the quantitative characteristic of the viral damage of monocytes. The decrease of the percentage of functionally active monocytes creates suitable conditions for the persistence of the virus in monocytes and for the disease to become chronic.     

The photoprotein pholasin as a luminescence substrate for detection of superoxide anion radicals and myeloperoxidase activity in stimulated neutrophils

Pholasin, the photoprotein of the common piddock Pholas dactylus, emits an intense luminescence upon oxidation. The contribution of superoxide anion radicals and myeloperoxidase (MPO) to Pholasin luminescence in stimulated neutrophils was investigated. Data on Pholasin luminescence were compared with results of superoxide anion radical generation detected by the cytochrome c test as well as with the release of elastase and MPO. In N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulated neutrophils, most of the luminescence is caused by superoxide anion radicals, whereas MPO shows only a small effect as shown by coincubation with superoxide dismutase (SOD) as well as potassium cyanide (KCN), an inhibitor of MPO. However, both, O₂^- and MPO contribute to light emission in fMLP/cytochalasin B and phorbol myristoyl acetate (PMA) stimulated cells. Thus, the kinetics of O₂^- generation and MPO release can be very well detected by Pholasin luminescence in stimulated neutrophils.

Degranulation of azurophilic granules was assessed using an ELISA test kit for released MPO or detection of elastase activity with MeO-Suc-Ala-Ala-Pro-Val-p-nitroanilide in the supernatant of stimulated cells. Both approaches revealed concurrently similar results concerning the amount and kinetics of enzyme release with data of Pholasin luminescence. Both, cytochrome c measurements and Pholasin luminescence indicate that fMLP/cytochalasin B and PMA stimulated neutrophils produce more O₂^- than fMLP stimulated cells. Thus, Pholasin luminescence can be used to detect, sensitively and specifically, O₂^- production and MPO release from stimulated neutrophils.     

Interrelation between the activity of hepatitis, liver fibrosis and immune status in children with chronic hepatitis B and C

The lesion of the liver in viral hepatitis was found to depend on the state of the immune system. Relationship between the content of lymphocyte subpopulations (CD3+, CD4+, CD8+, CD20+) in the blood and immunoglobulins (IgG, IgM, IgA) with parameters of semi-quantitative evaluation of the activity of hepatitis and the stage of liver fibrosis in children with chronic virus hepatitis B, C, B + C was studied. The characteristic feature of all hepatitis was a decrease in the number of T lymphocytes CD4+ below the normal level and an increase in the content of B lymphocytes. The correlation between the morphological activity of hepatitis and the amount of T lymphocytes CD8+ was established only in chronic hepatitis B. In chronic hepatitis B and B + C the absolute amount of blood lymphocytes decreased with the increase of the age of the patients, but in chronic hepatitis B this was accompanied by the decrease of the morphological activity of hepatitis and in hepatitis B + C by its increase. The amount of lymphocytes CD4+ rose with the increase of liver fibrosis in chronic hepatitis B. In children with chronic hepatitis C and B + C the amount of blood lymphocytes was found to be unrelated to the morphological activity of hepatitis.     

Cytochemical indices of leukocytes in patients with diabetes mellitus

Cytochemical indices of leukocytes were determined in 16 patients with diabetes mellitus in the period of unbalancing and balancing. The following tests were made: content of glycogen and lipids, acid phosphatase (AP), alkaline phosphatase (AIP), myeloperoxidase (MPO) and nonspecific alpha-naphtol acetate esterase (NANAE) activity. In unbalanced diabetics an evident decrease in the activity of AP and MPO could be noted as well as a decrease of glycogen content and an increase of lipid content. An insignificant decrease could be observed in the activity of ALP and NANAE in granulocytes. A slight increase in the activity of NANAE in monocytes would be found. Balancing this disease induced the increase of all parameters in granulocytes except MPO activity. It is interesting to note that balancing diabetes mellitus deepened the observed changes in the decrease or increase of tested parameters. The presented findings clearly indicate the role of metabolic disorders in diabetes mellitus on the activity of some neutrophilic enzymes and the glycogen and the content of lipids in neutrophils.     

The prevalence of hepatitis B and C markers among narcotic users

611 patients with acute parenteral virus hepatitis (VH) were studied with a view to find out markers indicating the presence of hepatitis B and hepatitis C virus infection (HB, HC, HB + C, HC + HBsAg). Of these, 166 patients (27.2%) systematically used narcotic drugs intravenously. Essential differences between drug users and VH patients without drug addiction were established regarding the distribution of patients by age and sex, the etiological structure and severity of the disease. Thus, in the group of drug users the prevalence of males, young people (15-29 years of age) and the mixed form of hepatitis B + C was noted. In VH patients using drug the disease took a more severe course than in such patients without drug addiction. The highest proportion of intravenously drug users with a severe and moderate course of the disease was found among patients with HB + C and HB.     

Age-related characteristics of the activity of phagocytes in the normal state and in infections

Activity of peroxidase (AP) and maintenance of lysosomal cation proteins (CP) were studied during phagocytic reaction of polymorphonuclear leukocytes of healthy persons of different age and those with acute respiratory disease and chronic tonsillitis. It is found that during acute virus and chronic bacterial infection similar changes in AP and CP dynamics are observed, which is correlated with a decrease of phagocytic digesting function.     

Neutrophil activation in vitro

Kinetics of spontaneous and induced reactions of Nitroblue tetrazolium (NBT) restoration by neutrophils was investigated in vitro. The index of activated neutrophils (IAN), commonly characterizing cell activation degree depending on the reaction time (t), was described by the equation: IAN = [NBT0](1-exp[-k1t]) + k0t; where (NBT0) is intracellular concentration of NBT, which reflects membrane permeability, k1 and k0--speed constants of the first and zero order reactions, characterizing, respectively, stimulation and spontaneous activities of enzymatic systems involved in NBT restoration. We detected the ability of heparin (2.0-7.5 mkg/ml) or chondrotin sulphate (0.25-2.5 mkg/ml) to activate neutrophils and presumably to maintain in vivo their spontaneous activity. The values of kinetics equation parameters enable us to speculate about relative hyper-, hypo- and normal function of neutrophils, related to the functional state of the whole organism. The value of [NBT0] changes from 8 to 103 conventional units, and a high correlation exists between this index and the reaction speed constants, differing in stimulated and spontaneously activated cells. The latter are represented by neutrophils only, which, as shown before, produce active forms of oxygen, used, probably, not for phagocytosis needs. The share of such cells makes approximately 30-40% of the neutrophil recirculating pool, and positively correlates with [NBT0], (r = 0.58; P < 0.05). This is determined by a very high membrane permeability of this neutrophil subpopulation. The detected different spontaneous and stimulated ability, various membrane permeability and enzyme system activity confirm the neutrophil functional unequality, which defines their activation peculiarity.     

The photoprotein pholasin as a luminescence substrate for detection of superoxide anion radicals and myeloperoxidase activity in stimulated neutrophils

Pholasin, the photoprotein of the common piddock Pholas dactylus, emits an intense luminescence upon oxidation. The contribution of superoxide anion radicals and myeloperoxidase (MPO) to Pholasin luminescence in stimulated neutrophils was investigated. Data on Pholasin luminescence were compared with results of superoxide anion radical generation detected by the cytochrome c test as well as with the release of elastase and MPO. In N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulated neutrophils, most of the luminescence is caused by superoxide anion radicals, whereas MPO shows only a small effect as shown by coincubation with superoxide dismutase (SOD) as well as potassium cyanide (KCN), an inhibitor of MPO. However, both, O₂^- and MPO contribute to light emission in fMLP/cytochalasin B and phorbol myristoyl acetate (PMA) stimulated cells. Thus, the kinetics of O₂^- generation and MPO release can be very well detected by Pholasin luminescence in stimulated neutrophils.

Degranulation of azurophilic granules was assessed using an ELISA test kit for released MPO or detection of elastase activity with MeO-Suc-Ala-Ala-Pro-Val-p-nitroanilide in the supernatant of stimulated cells. Both approaches revealed concurrently similar results concerning the amount and kinetics of enzyme release with data of Pholasin luminescence. Both, cytochrome c measurements and Pholasin luminescence indicate that fMLP/cytochalasin B and PMA stimulated neutrophils produce more O₂^- than fMLP stimulated cells. Thus, Pholasin luminescence can be used to detect, sensitively and specifically, O₂^- production and MPO release from stimulated neutrophils.     

寡核苷酸芯片法、实时PCR 和测序法进行乙肝病毒 基因分型的比较

目的:比较寡核苷酸芯片法、实时荧光PCR和测序法在对慢性乙肝患者病毒基因分型的比较和方法学评价。方法:对126例不同基因型的慢性乙肝患者的血清样本分别用寡核苷酸芯片法、实时荧光PCR法和测序法进行基因分型,并评价各种方法的临床表现、所需时间和检测成本。结果:寡核苷酸芯片法、实时荧光PCR分别能检测到1%和0.1%比例的基因型。在126例慢性乙肝患者的临床样本中,寡核苷酸芯片法、实时荧光PCR和测序法分别检测出41(33%)、41(33%)和45(36%)例为B型,76(60%)、76(60%)、81(64%)例为C型。寡核苷酸芯片法、实时荧光PCR均检出9例B、C混合基因型。在三种检测方法中实时荧光PCR是最快速和廉价的。结论:寡核苷酸芯片法、实时荧光PCR能检出B、C混合基因型,而测序法只能检测出样本的主导基因型。     

Functional properties of lymphocyte subpopulations in hepatitis B virus infection. I. Suppressor cell control of T lymphocyte responsiveness

Hepatocellular injury in hepatitis B virus infection may be produced by an autoaggressive hepatocytotoxic immune response. To test the hypothesis that acquired suppressor cell defects may participate in such a response, we assessed the functional integrity of 2 suppressor cell populations in patients with type B viral hepatitis. Spontaneous suppression of the 1-way mixed lymphocyte response by radiation-resistant, adherent peripheral blood mononuclear cells decreases during the acute phase of disease, returns towards normal with clinical recovery, but remains depressed in patients with chronic hepatitis. The degree of spontaneous suppressor cell dysfunction correlates inversely with at least 1 biochemical parameter of hepatocellular injury (SGPT). The functional integrity of this suppressor cell fluctuates during chronic hepatitis and may reflect currently undefined biologic variables in this disease. Mitogen-induced suppression on lymphocyte activation by radiation resistant, nonadherent suppressor cells is also depressed in acute and chronic hepatitis, but it does not correlate with biochemical evidence of hepatocellular injury on an individual-patient basis. Documentation of these generalized defects of nonspecific suppressor cell function establishes a basis for the possible existence of specific anomalies of immuno-regulation that may permit the expression of normally suppressed auoaggressive hepatocytotoxic immune mechanisms in viral hepatitis.     

Prevalence of Hepatitis δ (Delta) Virus Infection A Seroepidemiologic Study

In assessing the prevalence of hepatitis δ (delta) virus (HDV) infection in 358 patients with acute hepatitis B seen in Los Angeles between 1983 and 1985 and in 196 patients with chronic hepatitis B followed between 1980 and 1985, we found that 23% of patients with chronic and 5% of patients with acute hepatitis B were infected with HDV. Among patients with chronic hepatitis B, the prevalence of HDV infection was 73% in intravenous drug users and 14% in homosexual men. Acute coinfection with the hepatitis B virus was also more frequent in drug users (8%) than in other groups. δ-Hepatitis is a common infection in hepatitis B virus carriers in Los Angeles, particularly in drug addicts, but also in homosexual men who do not abuse drugs intravenously.     

Characteristics of the functional activity of human blood neutrophils using the nitroblue tetrazolium reduction test]

A study was made of the test of restoration of the nitroblue tetrazolium (NBT) by the intact blood neutrophils and the neutrophils stimulated with various endotoxin doses of Gram-negative bacteria in 70 healthy persons. A high sensitivity of the test with the NBT for the quantitative assessment of the neutrophil response under conditions stimulating the phagocytic activity was demonstrated. A functional nonhomogeneity of the neutrophil population, the most distinct with the minimal stimulating action, was noted.     

Effect of concurrent acute infection with hepatitis C virus on acute hepatitis B virus infection.

OBJECTIVE--To investigate the possible interference with acute hepatitis B virus infection by co-infection with hepatitis C virus. DESIGN--Analysis of stored sera collected for transfusion transmitted viruses study in 1970s. SETTING--Four major medical centres in the United States. PATIENTS--12 recipients of blood infected with hepatitis B virus. MAIN OUTCOME MEASURES--In 1970s, presence of antibodies in hepatitis B virus and raised serum alanine aminotransferase concentration; detection of antibodies to hepatitis C virus with new enzyme linked immunoassays. RESULTS--Five of the 12 patients were coinfected with hepatitis C virus. Hepatitis B surface antigen was first detected at day 59 in patients infected with hepatitis B virus alone and at day 97 in those coinfected with hepatitis C virus (p = 0.01); median durations of antigenaemia were 83 and 21 days respectively (p = 0.05), and the antigen concentration was lower in the coinfected patients. Alanine aminotransferase patterns were uniphasic when hepatitis B virus infection occurred alone (range 479-2465 IU/l) and biphasic in patients with combined acute infection (no value > 380 IU/l; p = 0.0025). Four coinfected recipients developed chronic hepatitis C virus infection. The fifth patient was followed for only four months. CONCLUSIONS--Acute coinfection with hepatitis C virus and hepatitis B virus inhibits hepatitis B virus infection in humans, and onset of hepatitis B may reduce the severity of hepatitis C virus infection but not frequency of chronicity. Alanine aminotransferase concentration showed a biphasic pattern in dual infection.     

Changes in the activity of the natural killers under the influence of genetically engineered alpha 2-interferon in patients with viral hepatitis B

The functional activity of natural killer cells (NKC) in 90 patients with acute viral hepatitis B was studied. As a result, the importance of this characteristic as a criterion of effectiveness of alpha 2-interferon obtained by gene engineering techniques was shown. The study revealed the presence of inverse relationship between the level of the functional activity of NKC and the severity of acute viral hepatitis B at the peak of the disease. The character of the influence of alpha 2-interferon on the cytotoxicity of NKC depended on the time of the use of the preparation. Administration of reaferon till day 7 of jaundice promoted a significant increase of the initially low activity of NKC in comparison with that in patients receiving common therapy. This was accompanied by rapid changes of the clinical signs of the disease and accelerated elimination of the virus from the blood. When the preparation was administered after day 6 of jaundice the activity of NKC increased slowly and only at the period of convalescence. These results recommend measurements of NKC activity as a criterion for the evaluation of the effectiveness of alpha 2-interferon.