[New Zwitter-Ionic Oligonucleotides: Preparation and Complementary Binding].

New zwitter-ionic oligonucleotide derivatives containing 1,2,3,4-tetrahydroisoquinoline-7-sulfonyl phosphoramidate group are described. Automated synthesis of these compounds was carried out according to the ß-cyanoethyl phosphoramidite scheme via the Staudinger reaction between 2-trifluoroacetyl-1,2,3,4-tetrahydroisoquinoline-7-sulfonyl azide and phosphite triester within oligonucleotide grafted to polymer support. 1,2,3,4-Tetrahydroisoquinoline-7-sulfonyl phosphoramidate group (THIQ) was stable under the conditions of standard oligonucleotide synthesis, including the removal of protective groups and cleavage of the oligonucleotide from the polymer support by treatment with a mixture of concentrated aqueous solutions of ammonia and methylamine (1 : 1) at 55°C. Oligonucleotides modified by one to five THIQ groups in various positions were obtained. The zwitter-ionic character of the obtained derivatives was reflected in their varying mobility under conditions of denaturing PAGE. The thermal stability of the duplexes of oligodeoxynucleotides containing THIQ groups with complementary DNA and RNA only slightly differed from that of natural DNA:DNA and DNA:RNA duplexes. The results reported suggest that oligonucleotides modified with zwitter-ionic THIQ groups as antisense therapeutic agents.

[Application of autofluorescence microscopy and laser Induced fluorescence methods to study the dynamics of the demineralization primary teeth process in vitro]. / Primenenie autofluorestsentnoi mikroskopii i lazernoi fluometrii dlya izucheniya protsessa demineralizatsii vremennykh zubov in vitro.

OBJECTIVE: Improvement of methods for studying the processes of demineralization of hard tissues of temporary teeth. MATERIAL AND METHODS: The study included primaries second molars (n=11). Samples of primary teeth were placed in a test tube with a demineralizing solution for - 1, 4, 8, 21 and 31 days. The of primary teeth samples were examined using methods - laser induced fluorescence (LIF) and autofluorescence microscopy (AFM). Assessment of the degree of demineralization of samples of temporary teeth was carried out according to the score scale developed by us. RESULTS: The enamel of the samples is demineralized slowly and evenly for up to 8 days with minimal objective signs, starting from the 8th day of the experiment, there is a significant increase in demineralization indicators. By the 21st day, the peak of demineralization is reached with partial dissolution of the enamel, an increase in the fluorescence effect to 80 UE, and reaches a maximum of 4 points on the evaluation scale. Dentin's hard tissues are demineralized gradually without "sudden jumps" in the fluorescence effect and at the same rate throughout the experiment, reaching a maximum on 31 days (30 UE - LIF). Dentin demineralization is characterized by less dissolution, however, the phenomenon of delamination is determined by the type of exfoliation of the organic dentin matrix, starting from the 21st day of the experiment. CONCLUSION: Enamel and dentin of deciduous teeth demineralize at different rates and have a characteristic specificity of morphological changes. Logistic regression analysis showed the consistency of the classifier for the predictive accuracy of each unit of the proposed scale for assessing the degree of demineralization of temporary teeth samples.

[Ultrasound anatomy of masticatory muscles. (Experimental and clinical study)]. / Ul'trazvukovaya anatomiya zhevatel'nykh myshts. (Eksperimental'no-klinicheskoe issledovanie).

PURPOSE: To describe the ultrasound anatomy of the masticatory muscles based on a comparison of the results of macroscopic and ultrasound studies of the masticatory muscles in vitro. MATERIAL AND METHODS: In the experimental part, a macropreparation of the masticatory muscle was studied; in a clinical example, an ultrasound of the masticatory muscles was performed on a patient with normal occlusion using the author's methods for analyzing echograms. RESULTS: The ultrasound anatomy of the masticatory muscles is described based on a comparison of data from the study of a macroscopic specimen and ultrasound images of the masticatory muscle of an experimental animal. Using a clinical example of a patient with normal occlusion, the results demonstrate the usage of the authors' developments in describing ultrasound images of the masticatory muscles. CONCLUSION: The study resulted in an algorithm for describing ultrasound images of masticatory muscles, including methods for qualitative and quantitative assessment of ultrasound images using proprietary developments involving elements of artificial intelligence technologies.

Mesyl phosphoramidate antisense oligonucleotides as an alternative to phosphorothioates with improved biochemical and biological properties.

Here we describe a DNA analog in which the mesyl (methanesulfonyl) phosphoramidate group is substituted for the natural phosphodiester group at each internucleotidic position. The oligomers show significant advantages over the often-used DNA phosphorothioates in RNA-binding affinity, nuclease stability, and specificity of their antisense action, which involves activation of cellular RNase H enzyme for hybridization-directed RNA cleavage. Biological activity of the oligonucleotide analog was demonstrated with respect to pro-oncogenic miR-21. A 22-nt anti-miR-21 mesyl phosphoramidate oligodeoxynucleotide specifically decreased the miR-21 level in melanoma B16 cells, induced apoptosis, reduced proliferation, and impeded migration of tumor cells, showing superiority over isosequential phosphorothioate oligodeoxynucleotide in the specificity of its biological effect. Lower overall toxicity compared with phosphorothioate and more efficient activation of RNase H are the key advantages of mesyl phosphoramidate oligonucleotides, which may represent a promising group of antisense therapeutic agents.

[Endoscopic treatment of choledocholithiasis in patients over 80 years old]. / Vozmozhnosti endoskopicheskogo lecheniya kholedokholitiaza u patsientov 80 let i starshe.

OBJECTIVE: To analyze endoscopic treatment of choledocholithiasis in patients over 80 years old. MATERIAL AND METHODS: A single-center retrospective study included 90 consecutive patients aged ≥80 years and 58 patients aged 60-79 years. Early outcomes including efficacy of calculus removal, incidence of complications and their risk factors were evaluated. RESULTS: In 75 patients aged ≥80 years (83.3%), endoscopic treatment was effective definitive single procedure. Between-group differences were insignificant (p=0.163). Patients aged ≥80 years were characterized by higher percentage of ASA classes 2-4 (p<0.001), age-adjusted mean Charlson comorbidity index (p=0.004), and ≥2 calculi (64.4% vs. 32.8%, p<0.001). Postoperative morbidity was similar (7.8% vs. 6.9%, p=0.842). Multivariate logistic regression analysis of potential risk factors did not reveal significant correlations: Chi-square test was 14.94 at 15 degrees of freedom (p=0.463). CONCLUSION: Similar postoperative morbidity determines safety of endoscopic lithoextraction in patients aged over 80 years. We should emphasize higher percentage of advanced age patients with «difficult¼ choledocholithiasis that requires appropriate training of specialists, adequate equipment of hospitals and routing of patients.

Phosphate-modif ied CpG oligonucleotides induce in vitro maturation of human myeloid dendritic cells.

Myeloid dendritic cells (DCs) play an important role in the immune response; therefore, the search for compounds that can effectively activate DCs is a needful goal. This study was aimed to investigate the effect of synthetic CpG oligodeoxynucleotides (CpG-ODN) on the maturation and allostimulatory activity of myeloid DCs in comparison with other PAMP and DAMP molecules. For the research, we synthesized known CpG-ODN class C (SD-101 and D-SL03) containing thiophosphate internucleotide groups, and their original phosphate-modified analogues (SD-101M and D- SL03M) with mesylphosphoramide internucleotide groups (M = µ-modification). The effects of CpG-ODN and other activators were evaluated on DCs generated from blood monocytes in the presence of GM-CSF and IFN-α (IFN-DC) or IL-4 (IL4-DC). Evaluation of the intracellular TLR-9 expression showed that both types of DCs (IFN-DC and IL4-DC) contained on average 52 and 80 % of TLR-9-positive cells, respectively. The CpG-ODNs studied enhanced the allostimulatory activity of IFN-DCs, and the effect of µ-modified CpG-ODNs was higher than that of CpG-ODNs with thiophosphate groups. The stimulating effect of CpG-ODN at a dose of 1.0 µg/ml was comparable (for D-SL03, D-SL03M, SD-101) with or exceeded (for SD-101M) the effect of LPS at a dose of 10 µg/ml. At the same time, IFN-DCs were characterized by greater sensitivity to the action of CpG-ODNs than IL4-DCs. The enhancement of DC allostimulatory activity in the presence of CpG-ODNs was associated with the induction of final DC maturation, which was confirmed by a significant decrease in the number of CD14+DC, an increase in mature CD83+DC and a trend towards an increase in CD86+DC. Interestingly, the characteristic ability of LPS to enhance the expression of the co-stimulatory molecule OX40L on DCs was revealed only for the µ-analogue SD-101M. In addition, CpG-ODNs (SD-101 and SD-101M) had a stimulatory effect on IFN-γ production comparable to the action of LPS. The data obtained indicate a stimulating effect of CpG-ODN on the maturation and allostimulatory activity of human myeloid DCs, which is more pronounced for µ-modified analogs.

Two-component 10-23 DNA enzymes.

A new strategy for engineering of catalytic two-component constructions based on 10-23 DNAzyme was proposed. The using of a combination of shortened DNAzyme with 2'-O-methyl oligomers as effectors significantly increased the catalytic activity of this DNAzyme.

Effects of enkephalins and their analogs on gastric secretion.

The effects of enkephalin analogs on gastric secretion were studied. It was found that gastric secretion stimulated by histamine was significantly increased under the influence of the enkephalin analog Tyr-D-Ala-Gly-Phe-(NO2)NH2. The enkephalin analog Tyr-Gly-Gly-Phe-Leu-D-Arg increased the amount of gastric juice and the secretion of pepsin, but the hydrochloric acid debit was lower than in experiments with perfusion of histamine alone. Another enkephalin analog, Tyr-D-Ala-Gly-Phe-D-Leu-Arg, introduced simultaneously with histamine, significantly lowered the secretion of gastric juice only 90-105 min after the beginning of perfusion.

There are receptors for neurotensin in the caudate nucleus and posterior hypothalamus.

It was established in chronic experiments on dogs that the introduction of neurotensin in a dose of 3 or 10 micrograms in 1.5 microliters into the caudate nucleus changes the parameters of the conditioned and unconditioned food reflexes: a 31% shortening of the latent period and a 56% increase in the magnitude of the reflex. Microapplication of neurotensin in the same doses in the posterior region of the hypothalamus enhanced the secretory function of the gland caused by the introduction of histamine. It was concluded that the caudate nucleus and hypothalamus contain cells possessing receptors for neurotensin, which participate in mechanisms of formation of the conditioned reflex and central influence on the organs of the digestive system.

[Concentration of gastrin in serum, gastric and duodenal tissues and gastric argentaffin cells following administration of pancreozymin and glucagon]. / Soderzhanie gastrina v syvorotke krovi, tkaniakh zheludka, dvenadtsatiperstnoi kishki krys i argentaffinnye kletki zheludka posle vozdeistviia pankreoziminom i gliukagonom.

In acute experiments on rats, pancreozimine and glycagon increased 2-3 fold the gastrin contents in the blood serum and duodenum tissues. Within 15 min of glycagon administration the gastrin contents increased 2 fold and then reduced in the mucosa of the stomach antral portion. Administration of pancreozimine elicited the opposite effect. After the action of these hormones the amount of argentaffine cells obviously decreased in the stomach antral portion.

[Action of enkephalins and their analogs on gastric secretion]. / Deistvie énkefalinov i ikh analogov na zheludochnuiu sekretsiiu.

The gastric secretion induced by histamine increased under the influence of enkephalin analog Tyr--D--Ala--Gly--Phe(NO2)NH2. The enkephalin analog Tyr--Gly--Gly--Phe--Leu--D--Arg increased the amount of gastric juice and pepsin, but the debit of hydrochloric acid was lower than in perfusion of histamine alone. Another analog of enkephalin Tyr--D--Ala--Gly--Phe--D--Leu--Arg administered together with histamine decreased the gastric juice secretion only within 90--105 min after the beginning of the perfusion.

[Effect of neurotensin and (DTr11)-neurotensin on the secretory function of the pancreas]. / Deistvie neirotenzina i (DTr11)-neirotenzina na sekretornuiu funktsiiu podzheludochnoi zhelezy.

In chronic experiments on dogs, administration of neurotensin into cerebral structures (hypothalamus, caudate nucleus) or into the blood flow exerted the same effect: enhancement of the pancreas secretory function, depending on the dosage of the i.v. administration. In contrast, peripheral administration of neurotensin analogue: the DTr11-neurotensin inhibited the pancreas secretory function. Administration of the DTr11-neurotensin into the cerebral structures increased the secretory activity of the pancreas. The hypothalamus and the caudate nucleus seem to have receptors sensitive to neurotensin and DTr11-neurotensin.

[Existence of neurotensin receptors in the caudate nucleus and posterior hypothalamus]. / V khvostatom iadre i zadnem gipotalamuse est' retseptory k neirotenzinu.

Chronic experiments in dogs revealed that administration of neurotensin (3 and 10 mkg in 1.5 mkl) into the caudate nucleus altered the parameters of conditioned and unconditioned food reflexes: shortened the latency by 31% and augmented the reflexes by 56%. The microapplication of neurotensin in the same dosage into the posterior hypothalamus increased the stomach secretory function induced by histamine administration. The caudate nucleus and the hypothalamus seem to contain cells which have the receptors for neurotensin and take part in the mechanisms of conditioning and central influences on the digestive system's organs.

[Role of antral gastrin in regulating gastric secretion and bile flow]. / Rol' antral'nogo gastrina v reguliatsii zheludochnoi sekretsii i zhelchevydelenii.

In chronic experiments on dogs with removed mucosa of the antral portion of the stomach, administration of the liver extract into the stomach or intestine decreased the output of hydrochloric acid as compared with intract animals. The liver extract administration caused no raise of gastrin level in the blood serum of the operated animals.