[Late neurosyphilis in a psychiatric hospital]. / Pozdnii neirosifilis v psikhiatricheskom statsionare.

AIM: The problem of detection and diagnosis of late forms of neurosyphilis remains relevant and to a large extent concerns the field of psychiatry. The autors study of professional activity of psychiatric hospital in this field with use of clinical and statistical data. MATERIAL AND METHODS: The article presents modern statistical data on the detection of these forms of syphilis in psychiatric hospitals of the Moscow region in comparison with the indicators of 2008-2010. RESULTS AND CONCLUSION: There were 2 clinical cases of late neurosyphilis identified and received specific therapy in a psychiatric hospital in 2016. It is concluded that there is a need for a cerebrospinal fluid analysis of all seropositive patients in psychiatric hospitals, including those who received treatment for various forms of syphilis.

Three cases of late syphilitic hepatitis.

The article presents literature data for the last 10 years on specific liver damage. There are three own clinical cases of syphilitic hepatitis, one of them - with a lethal outcome. Attention is drawn to the importance of early diagnosis of syphilitic hepatitis. The main criteria for the diagnosis of this pathology.

Validation and development of MTH1 inhibitors for treatment of cancer.

BACKGROUND: Previously, we showed cancer cells rely on the MTH1 protein to prevent incorporation of otherwise deadly oxidised nucleotides into DNA and we developed MTH1 inhibitors which selectively kill cancer cells. Recently, several new and potent inhibitors of MTH1 were demonstrated to be non-toxic to cancer cells, challenging the utility of MTH1 inhibition as a target for cancer treatment. MATERIAL AND METHODS: Human cancer cell lines were exposed in vitro to MTH1 inhibitors or depleted of MTH1 by siRNA or shRNA. 8-oxodG was measured by immunostaining and modified comet assay. Thermal Proteome profiling, proteomics, cellular thermal shift assays, kinase and CEREP panel were used for target engagement, mode of action and selectivity investigations of MTH1 inhibitors. Effect of MTH1 inhibition on tumour growth was explored in BRAF V600E-mutated malignant melanoma patient derived xenograft and human colon cancer SW480 and HCT116 xenograft models. RESULTS: Here, we demonstrate that recently described MTH1 inhibitors, which fail to kill cancer cells, also fail to introduce the toxic oxidized nucleotides into DNA. We also describe a new MTH1 inhibitor TH1579, (Karonudib), an analogue of TH588, which is a potent, selective MTH1 inhibitor with good oral availability and demonstrates excellent pharmacokinetic and anti-cancer properties in vivo. CONCLUSION: We demonstrate that in order to kill cancer cells MTH1 inhibitors must also introduce oxidized nucleotides into DNA. Furthermore, we describe TH1579 as a best-in-class MTH1 inhibitor, which we expect to be useful in order to further validate the MTH1 inhibitor concept.

[Awareness of knowledge of faces of the preinduction age on problems of sexually-transmitted infections].

The necessity for examination of the level of youth awareness about the ways of transmission, safety products and prevention of sexually-transmitted infections is approved. It is recommended to begin the sexual education of scholars since 11-12 years. Development of the educational programs must be based on results of analysis of systematic questionnaire of scholars.

Structure of Cry3A delta-endotoxin within phospholipid membranes.

Interaction of delta-endotoxin and its proteolytic fragments with phospholipid vesicles was studied using electron microscopy, scanning microcalorimetry, and limited proteolysis. It was shown that native protein destroys liposomes. The removal of 4 N-terminal alpha-helices and the extreme 56 C-terminal amino acid residues did not affect this ability. The results obtained by limited proteolysis of delta-endotoxin bound to lipid vesicles show essential conformational changes in three or four N-terminal helices and in the C-terminal region. The calorimetric method used in this study provides a unique possibility for the validation of existing models of protein binding and for a more accurate determination of the regions where conformational changes take place. It was found that the binding of the protein to model liposomes does not alter its structure in the regions starting with the fourth alpha-helix of domain I. This can be concluded from the fact that the activation energy of denaturation of the protein remains unchanged upon its binding to the phospholipid membranes. A new structural model has been proposed which agrees with the data obtained.

[Characteristic of mutants of Pseudomonas putida IPM-36 recombinant strains, selected by anticipating growth on a media containing an inducer of cry3A gene expression]. / Kharakteristika mutantov rekombinantnogo shtamma Pseudomonas putida IPM-36, otobrannykh po priznaku operezhaiushchego rosta na srede s induktorom ékspressii cry3A-gena.

Induction of the expression of the delta-endotoxin gene from Bacillus thuringiensis var. tenebrionis in the recombinant strain Pseudomonas putida IPM-36 negatively affected the viability and the growth rate of the culture. In order to optimize the insecticide production by the recombinant strain, mutant clones exhibiting anticipating growth on an inducer-containing medium were selected and studied. These clones differed in such aspects as the localization of mutations (either in plasmid pBTN11, carrying the cry3A gene, or in the chromosome), growth rate, or the level of delta-endotoxin synthesis after induction. Several mutants obtained proved much superior to P. putida IPM-36 in their structural and segregation stability, although they were as efficient as the original strain with respect to the production of the insecticide (protei Cry3A).

Transition state of the rate-limiting step of heat denaturation of Cry3A delta-endotoxin.

Heat denaturation of Cry3A delta-endotoxin from Bacillus thuringiensis var. tenebrionis and its 55 kDa fragment was studied by differential scanning microcalorimetry at low pH. Analysis of the calorimetric data has shown that denaturation of Cry3A delta-endotoxin is a nonequilibrium process at heating rates from 0. 125 to 2 K/min. This means that the stability of delta-endotoxin (the apparent temperature of denaturation Tm) under these conditions is under kinetic control rather than under thermodynamic control. It has been shown that heat denaturation of this protein is a one-step kinetic process. The enthalpy of the process and its activation energy were measured as functions of temperature. The data obtained allow confirmation of the fact that the conformation of delta-endotoxin at the transition state only slightly differs from its native conformation with respect to compactness and extent of hydration. The comparison of the activation energy for intact delta-endotoxin and the 55 kDa fragment showed that the transition of the molecule to a transition state does not cause any changes in the conformation of three N-terminal alpha-helices. Complete removal of the N-terminal domain of delta-endotoxin and 40 amino acids from the C-terminus beta-sheet domain III causes an irreversible loss of the tertiary structure. Thus, during protein folding the nucleation core determining protein stability does not involve its three initial alpha-helices but may include the remaining alpha-helices of the N-terminal domain. The functional significance of peculiarities of structure arrangement of the delta-endotoxin molecule is discussed.

[Features of the production of CryIIIA delta-endotoxin from Bacillus thuringiensis in a recombinant strain of Pseudomonas putida under control of Pm and xylS regulatory elements]. / Osobennosti produktsii CryIIIA delta-éndotoksina Bacillus thuringiensis v rekombinantnom shtamme Pseudomonas putida pod kontrolem reguliatornykh élementov Pm i xylS.

The addition of 3-methyl benzoate to the culture of the recombinant strain Pseudomonas putida IPM-36 (bearing the cryIIIA gene of B. thuringiensis subsp, tenebrionis under the control of the Pm promoter and the regulator gene xylS) slowed down the growth rate of the recombinant strain and increased, under non-selective conditions, the number of plasmid-free cells. Intense synthesis of the Coleoptera-specific delta-endotoxin encoded by the cryIIIA gene began 6-8 h after the addition of the inducer 3-methyl benzoate, no matter whether it was added in the early or late logarithmic phase. Maximal production of endotoxin (0.5-0.6 g/l) was observed when the inducer was added in the early logarithmic phase (3 h of growth). Overproduction of delta-endotoxin impaired cell division, so that filamentous cells became predominant in the culture. delta-Endotoxin accumulated in overproducing cells as irregular crystalloid inclusions.

Application of free-flow electrophoresis for isolation and purification of proteins and peptides.

Free-flow electrophoresis (FFE) has been applied to the separation and purification of a variety of proteins and polypeptides: bee venom, tumor necrosis factor, interleukin-1beta, interferon-gamma and superoxide dismutase. FFE at constant pH and conductivity of the carrying buffer is shown to be efficient at various separation schemes. In some cases, the method allows us to obtain proteins with a purity of more than 90% at a productivity of 20-30 mg/h. An electrophoretic apparatus with a new, multi-sectional construction of the electrophoretic chamber and a system for cross-displacement of carrying buffer in the chamber is described.

[Thermodynamic analysis of domain organization of Bacillus thuringiensis toxins]. / Termodinamicheskii analiz domennoi organizatsii toksinov Bacillus thuringiensis.

The structure of delta-endotoxins CryIA(c) and CryIIIA from Bacillus thuringiensis was studied by differential scanning microcalorimerty. The analysis of molecular melting showed that the N- and C-terminal halves of the CryIA(c) protoxin from B. thuringiensis subspecies kurstaki HD-73 are thermodynamically independent subunits, with the C-terminal fragment being denatured at a much lower temperature than the N-terminal fragment. The tertiary structure of the N-terminal fragment undergoes no changes during the protoxin-toxin transition. The melting of the native structure of CryIA(c) at pH 9.7-11.0 suggests that it consists of two domains. In CryIIIA from B. thuringiensis subspecies tenebrionis, the transition from the native to denatured state under alkaline conditions (pH 9.7-11.0) proceeds by the "two-state" principle; i.e., the protein melts as one cooperative domain. The melting of the CryIIIA toxin at pH 2.2-3.5 is described by two transitions overlapping by temperatures, indicating the presence of two domains.

[Treatment efficacy in patients with secondary recurrent and latent early syphilis based on a new abbreviated method]. / Effektivnost' lecheniia bol'nykh vtorichnym retsidivnym i skrytym rannim sifilisom po novomu ukorochennomu metodu.

The authors analyze the findings of clinical and serological follow-up of 370 patients with secondary relapsing and early latent syphilis treated with a new rapid method. This method consists in intramuscular injections of water-soluble penicillin in high daily doses (4,000,000, 6,000,000, 8,000,000 U) made for 3-4 weeks. The follow-up period of 1-6 years has confirmed a high therapeutic efficacy of this method.

Correlation of the character of intramolecular melting with digestibility by pepsin in precipitating and non-precipitating pig anti-Dnp antibodies.

Precipitating and non-precipitating pig anti-2,4-dinitrophenyl group (Dnp) antibodies were investigated by differential adiabatic scanning microcalorimetry in the pH range 3.7-4.5. The partial heat capacity functions obtained revealed a notable difference between the two antibody types when the analysis was done at pH 4.5 or 4.0. The transition observed at temps around 50 degrees C in a non-precipitating antibody was absent in a precipitating antibody. Under analogous conditions, pH 4.5, the precipitating antibody was fully resistant to pepsin, while the non-precipitating antibody yielded appropriate F(ab')2 and pFc'fragments. At pH 3.7 no substantial difference in the partial heat capacity function could be observed between the two antibody types. Below pH 4.0 the precipitating antibody became susceptible to peptic cleavage and yielded fragments of the same general character as the non-precipitating antibody. This finding lends support to the view that the structural block in immunoglobulin G that melts first, i.e. at temps near 50 degrees C, is the CH2 domain or a part of it.