Chrono- and Immunocorrection of Inflammatory Disorders of Internal Reproductive Organs in Women of Reproductive Age.

We compared the effectiveness of immunomodulators used in the treatment of patients with chronic salpingitis and oophoritis with or without changes in succinate dehydrogenase (SDH) activity in blood lymphocytes at incubation with the drug. Diurnal variations in individual reaction of SDH in blood lymphocytes to thymalin or ridostin were revealed. In the groups of women receiving ridostin or thymalin during the reaction of lymphocyte SDH to it, improvement of clinical laboratory and immunological parameters was observed in the majority of the patients and no effect was found in a lesser group of patients than in the groups treated with drugs during the absence of lymphocyte SDH reaction thereto. The timing of the presence of SDH reaction to drugs in the immunocompetent cells makes it possible to set the optimal daily regime of their application and to select a drug that would be most effective in each particular case.

Hypouricemic effects of novel concentrative nucleoside transporter 2 inhibitors through suppressing intestinal absorption of purine nucleosides.

We have developed concentrative nucleoside transporter 2 (CNT2) inhibitors as a novel pharmacological approach for improving hyperuricemia by inhibiting intestinal absorption of purines. Dietary purine nucleosides are absorbed in the small intestines by CNTs expressed in the apical membrane. In humans, the absorbed purine nucleosides are rapidly degraded to their final end product, uric acid, by xanthine oxidase. Based on the expression profile of human CNTs in digestive tract tissues, we established a working hypothesis that mainly CNT2 contributes to the intestinal absorption of purine nucleosides. In order to confirm this possibility, we developed CNT2 inhibitors and found that (2R,3R,4S,5R)-2-(6-amino-8-{[3'-(3-aminopropoxy)-biphenyl-4-ylmethyl]-amino}-9H-purin-9-yl)-5-hydroxymethyl-tetrahydrofuran-3,4-diol (KGO-2142) and 1-[3-(5-{[1-((2R,3R,4S,5R)-3,4-dihydroxy-5-hydroxymethyl-tetrahydrofuran-2-yl)-1H-benzimidazol-2-ylamino]-methyl}-2-ethoxyphenoxy)-propyl]-piperidine-4-carboxylic acid amide (KGO-2173) were inhibitory. These CNT2 inhibitors had potent inhibitory activity against inosine uptake via human CNT2, but they did not potently interfere with nucleoside uptake via human CNT1, CNT3 or equilibrative nucleoside transporters (ENTs) in vitro. After oral administration of KGO-2173 along with [(14)C]-inosine, KGO-2173 significantly decreased the urinary excretion of radioactivity at 6 and 24h in rats. Since dietary purine nucleosides are not utilized in the body and are excreted into the urine rapidly, this decrease in radioactivity in the urine represented the inhibitory activity of KGO-2173 toward the absorption of [(14)C]-inosine in the small intestines. KGO-2142 almost completely inhibited dietary RNA-induced hyperuricemia and the increase in urinary excretion of uric acid in cebus monkeys. These novel CNT2 inhibitors, KGO-2142 and KGO-2173, could be useful therapeutic options for the treatment of hyperuricemia.

[Efficacy of combined administration of ridostin and oseltamivir (tamiflu) for therapy and prophylaxis of experimental infection induced by influenza virus A (H5N1) in mice].

AIM: Study of possibility of treatment-prophylaxis effect increase during combined administration of ridostin and tamiflu in experiments in mice infected with highly pathogenic influenza virus strain A/chicken/Kurgan/05/2005 (H5N1). MATERIALS AND METHODS: Balb/c line mice infected intranasally with influenza virus at 100 and 10 LD50 doses received ridostin and tamiflu as monopreparation or the combined variant before or after the infection. The mice were observed for 16 days, lethality rate, protection coefficient and average life span were evaluated. Virus concentration in lungs was determined by using titration in MDCK cell line. RESULTS: Combined administration ofridostin and tamiflu after the infection increased survivability of the animals when compared with the control group, and reduced influenza virus concentration in lungs. CONCLUSION: Treatment effect during combined administration of ridostin and tamiflu after influenza virus infection increased.

[Biological properties of a prolonged action interferon inductor].

AIM: Evaluation of composite formulation of yeast double stranded RNA with polyglucinum (dsRNA-PG) effect on non-specific antiviral resistance factors in mice in comparison with commercial formulation Ridostin. MATERIALS AND METHODS: dsRNA and Ridostin formulations were injected intramuscularly once at the dose of 5 mg/ml, polyglucinum--at the dose of 3.75 mg/ml. 3, 5, 24, 48 and 72 hours after the injection serum interferon levels, neutrophil oxidation-reduction activity parameters, peritoneal macrophage phagocyte activity levels were analyzed in mice blood samples. RESULTS: New dsRNA and polyglucinum containing composite formulation is a non-specific resistance system stimulator. dsRNA-PG effect on interferon synthesis and mice phagocyte activity was higher than with Ridostin and developed earlier. Neutrophil function activation by the formulation had a prolonged effect. A possible explanation for increased activity of dsRNA and polyglucinum composite formulation is a modulating effect by the polysaccharide component. CONCLUSION: The new formulation may have a more intensive and prolonged protective effect against influenza virus in comparison with Ridostin.

[Estimation of possible use of interferon inductors for prevention of postvaccional reactions to TEOVac in animal models].

The experiments on guinea pigs showed that arbidol administered orally in a single dose 24 hours prior vaccination with TEOVAC and ridostin administered in a single dose intranasally on the 4th day after the vaccination lowered the vaccine virus accumulation in the animal organs and tissue without any effect on the vaccine immunogeneity. The results are someway indicative of the possible use of the interferon inductors for prevention of postvaccinal reactions to TEOVAC.

[Allergenic and immunotropic properties of yeast RNA-tilorone hydrochloride molecular complex].

Immunotropic properties of the interferon-inducing molecular complex (MC) yeast RNA--tilorone hydrochloride have been under study. MC was experimentally studied in vivo to establish its influence on the amount of antibody-forming cells and the level of antibody formation. The influence of MC on the oxygen-generating activity of spleen macrophages was established in the HCT test. MC in a dose of 1.25 mg/kg was shown to considerably activate immunocompetent cells, thus producing pronounced influence on humoral immunity. In addition, the study showed the dose dependence of the influence of MC on individual elements of the immune system as well as differences in the dynamics of immunomodulation caused by the use of high and low doses of MC. The data thus obtained made it possible to regard MC as a promising immunomodulator.

Carry-over effects of dietary yeast RNA as a source of nucleotides on lymphoid organs and immune responses in Leghorn-type chickens.

1. The carry-over effects of supplementing Leghorn-type chickens with yeast RNA as a dietary source of nucleotides for 4 weeks on growth, lymphoid organ weights and immune responses were assessed in a 12-week study. 2. A commercial starter feed supplemented with 0 (control), 5 (LR) or 10 (HR) g yeast RNA/kg was offered to 1-d-old male ISA Brown chicks for 4 weeks, and then all birds were given a commercial pullet grower feed for another 8 weeks. Growth performance, antibody responses to sheep red blood cells (SRBC) and cutaneous reactivity of toe webs to phytohaemagglutinin (PHA)-M were measured at 4-week intervals. 3. Growth rates, feed intake and feed efficiency were not affected by dietary yeast RNA during the supplementary period, but birds previously offered the HR diet grew faster than control birds during weeks 4 to 8.4. LR-fed birds had a higher spleen weight relative to body weight (BW) than control birds at week 4, but this effect was not detected at other times. 5. Serum primary antibody levels against SRBC were not affected by dietary yeast RNA at any time. 6. The toe-web PHA response was significantly higher at week 8 in control birds than in birds previously given the LR diet, although no difference among dietary treatments was observed at other times. 7. It is concluded that the addition of yeast RNA as a source of nucleotides to a commercial diet selectively stimulated the development of the spleen in young birds, but this effect did not persist into a later stage of the bird's life.

[An intensification of antiviral and interferon effects of ridostin (an interferon inducer) by cationic liposomes in vitro and in intranasal administration]. / Usilenie kationnymi liposomami protivovirusnogo i interferonogennogo deistviia induktora interferona ridostina in vitro i pri intranazal'nom primenenii.

Combined application of ridostine with catonic liposomes was shown to essentially enhance the interferon-inducing and antiviral activity of the former in experiments with cell cultures L-929, which is apparently related with an improved efficiency of intracellular delivery of dsRNA. A comparative study demonstrated that ridostine, when combined with liposomes, is needed by 10(3)-10(4) times less as when it is used alone. A pretreatment of the cellular monolayer by cationic liposomes contributes also to enhancing the activity of ridostine, which can be explained by an enhanced permeability of cells for dsRNA holding on-for as long as 30 minutes after the removal of liposomes from the liquid culture. A separate successive administration of, first, liposomes and, then, of ridostine in BALB/c mice (20 mg/kg) leads to a more intensified induction of interferon in the upper respiratory tract tissues as compared with the administration of ridostine alone.

Purine derivative excretion in dairy cows: endogenous excretion and the effect of exogenous nucleic acid supply.

An experiment was conducted with dairy cows to study the partitioning of excreted purine derivatives between urine and milk and to quantify the endogenous contribution following the isotopic labeling of microbial purine bases. Three lactating cows in their second lactation that had been cannulated in the rumen and the duodenum were fed a mixed diet (48:52, roughage/concentrate ratio) distributed in equal fractions every 2 h, and duodenal flow of purine bases was determined by the dual-phase marker system. Nitrogen-15 was infused continuously into the rumen to label microbial purine bases, and the endogenous fraction was determined from the isotopic dilution in urinary purine derivatives. Urinary and milk recovery of duodenal purine bases were estimated at early (wk 10) and late (wk 33) lactation by the duodenal infusion of incremental doses (75 and 150 mmol purine bases/d) of RNA from Torula yeast. Each period was 6 d, with RNA being infused during the last 4 d, followed by measurement of the flow of purine bases to the duodenum. The isotope dilution of purine derivatives in urine samples confirmed the presence of an endogenous fraction (512 +/- 36.43 micromol/W0.75 or 56.86 mmol/d) amounting to 26 +/- 3.8% of total renal excretion. Total excretion of purine derivatives in urine plus milk was linearly related to the duodenal input of purine bases, but the slopes differed (P < 0.005) between lactation stages resulting in a lower equimolar recovery in early (y = 58.86 (+/-3.89) +0.56 (+/-0.0164) x; r = 0.90) than late lactation (y = 58.86 (+/-3.89) + 0.70 (+/-0.046) x; r = 0.80). Excretion of purine derivatives through milk represented a minimum fraction of total excretion but responded significantly to the duodenal input of purine bases. No differences between lactation stages were detected, and variations in milk yield did modify significantly the amount of purine derivatives excreted through the milk.

Correlations between in vitro effects of preparations of interferon and its inducers on blood cells in patients with multiple sclerosis.

We studied in vitro production of interferon-alpha and interferon-gamma by peripheral blood leukocytes from 15 patients with multiple sclerosis. The priming effects of interferon preparations weakly correlated with interferon-alpha production by leukocytes from patients with multiple sclerosis, but negatively correlated with interferon-gamma production. The effects of interferon inducers in most cases positively correlated with its spontaneous production. We found a weak positive correlation between the priming effect of natural interferon-alpha and the effect of recombinant interferons. There were positive or strong positive correlations between the effects of recombinant interferons on leukocytes from patients with multiple sclerosis. The relationship between the effects of medicinal interferon inducers and interferon preparation varied from negative to strong positive correlations. These data suggest that correlation analysis can be used for dynamic control and elaboration of methods for combined immunotherapy of multiple sclerosis with various interferon preparations or interferon and its inducers.

[Some aspects of interferon formation in white mice]. / Nekotorye aspekty interferonoobrazovaniia v organizme belykh myshei.

White mice weighing 14-16 g were intranasally infected with LD50 of influenza virus (A/Aichi/2/68 strain). High levels both of virus and interferon were detected in the lung. Sufficient virus accumulation in the nasal cavity occurred with low interferon induction. At the same time high blood interferon levels corresponded to sporadic low viremia. Intraperitoneal injection of the interferon inducer ridostin (a pharmacological formulation of dsRNA) to BALB/c mice (18-20 g) in a dose of 5 mg/kg induced intensive blood accumulation of interferon with its peak at 8 hours postadministration (2560 U/0.2 ml), but interferon was not detected in the respiratory tract and brain of these mice. Intranasal (15 mg/kg) and aerogenic (0.4-0.6 mg/kg) administration of ridostin induced interferon mainly in the upper respiratory tract and lung. The regularities found are in agreement with the data on interferon induction by other dsRNA preparations, which makes it necessary to design dosage forms of interferon inducers for respiratory application in influenza.

[Dynamics of interferon induction in albino mice by interferon inducer ridostin administered by various routes]. / Izuchenie dinamiki interferonoobrazovaniia v organizme belykh myshei pri razlichnykh putiakh vvedeniia induktora interferona ridostina.

The time dependence of interferon production in blood, tissues of the respiratory tract, brain and olfactory tract of mice BALB/c was investigated after administration of the interferon inductor ridostin by various routes. Intraperitoneal injection of ridostin in a dose of 5 mg/kg induced intensive accumulation of interferon in the blood serum with the peak in 8 hours (2560 U/0.2 ml) while no interferon was detected in the tissues of the respiratory tract and brain of the animals. Intracerebral injection of ridostin in the same dose induced accumulation of interferon in both the tissues of the brain (maximum 160 U/0.2 ml in 24 hours) and the blood serum (maximum 1280 U/0.2 ml in 8 hours). After respiratory administration of ridostin interferon was detected only in the site of the administration in the tissues of the upper respiratory tract and lungs of the mice.

[Features of formation of antiviral resistance upon local application of the interferon inducer ridostin]. / Osobennosti formirovaniia protivovirusnoi ustoichivosti pri mestnom primenenii induktora interferona ridostina.

Antiviral potency of an ointment dosage form of interferon (IF) inducer ridostin has been demonstrated in guinea pigs with genital herpes. Early beginning of therapy led to a 72% decrease of the intensity of clinical signs with cure 12-13 days earlier than in the control. Application of ridostin ointment onto the skin of intact animals caused an increase of IF level in the blood and of the phagocytic activity of peritoneal exudation macrophages. Ridostin effect on the total-systems factors of nonspecific defense appears to be one mechanism in the formation of antiviral resistance upon external application of this IF inducer.

[The antitumor action of double-stranded interferon inducers (larifan, ridostin) and reaferon in an experiment]. / Protivoopukholevoe deistvie dvuspiral'nykh induktorov interferona (larifana, ridostina) i reaferona v éksperimente.

Interferon inducers larifan and rhidostin, and reaferon were shown to exert an inhibiting antitumor effect manifested in the prolongation of the incubation period, decrease of the size of tumors, and longer survival of the animals. The maximal anti-tumor and immunomodulating effect was obtained by combined use of preimmunization with tumor cells and simultaneous administration of reaferon or interferon inducers, larifan and rhidostin. Larifan was also shown to have a greater antitumor activity than rhidostin. Larifan, however, was maximally active only in combination combination with vaccination using syngeneic cells of the virus-induced tumor. In this case levels of alpha and gamma interferons were 2-4 times higher than normally.

Immunohemopoietic effects of dietary nucleotide restriction in mice.

The influence of dietary sources of nucleotides on host in vivo and in vitro immuno-hematologic responses in BALB/c (NCI) mice was studied. Adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) were measured in popliteal lymph nodes undergoing proliferative response to syngeneic and allogeneic in vivo stimulation. Supplementation of a nucleotide-free (NF) diet with yeast RNA (NFR) or uracil (NFU) significantly enhanced the host PLN immune response as compared with NF and NF supplemented with adenine (NFA) diets. Levels of ADA and PNP enzymes in the PLNs increased with the alloimmune PLN response of host, and immunosuppression was associated with decreased ADA and PNP activities in lymphocytes following antigenic stimulation. The induction of these enzymes during immune response appears to require dietary sources of certain nucleotides. When bone marrow cells from control chow fed animals were cultured with supernatants (sups) from mitogen activated splenocytes of animals on each dietary group, NF sups significantly decreased (P less than 0.05) the BM proliferative response compared with the response observed with NFR sups, and similar to NFA or NFU sups. When stimulated with purified IL-3, NFR BM cells had higher levels of Thy1.2 or Lyt 1 surface markers as compared with other test groups. In the in vivo splenic colony formation-CFUs assay, spleens from NFR- and NFU-fed animals had a significantly higher number of colonies than spleens from NF- or NFA-fed mice. Thus, NF diet decreases both in vivo lymphoproliferation response to alloantigen and hemopoietic growth factor production, rendering the host splenic environment deficient for stem cell growth. These adverse effects are reversed by RNA supplementation of NF diet. These nutritional studies demonstrate a critical and regulatory role for dietary nucleotides in immunohemopoiesis.