TROPHY registry - status report.

INTRODUCTION: The TROPHY registry has been established to conduct an international multicenter prospective data collection on the surgical management of neonatal intraventricular hemorrhage (IVH)-related hydrocephalus to possibly contribute to future guidelines. The registry allows comparing the techniques established to treat hydrocephalus, such as external ventricular drainage (EVD), ventricular access device (VAD), ventricular subgaleal shunt (VSGS), and neuroendoscopic lavage (NEL). This first status report of the registry presents the results of the standard of care survey of participating centers assessed upon online registration. METHODS: On the standard of treatment forms, each center indicated the institutional protocol of interventions performed for neonatal post-hemorrhagic hydrocephalus (nPHH) for a time period of 2 years (Y1 and Y2) before starting the active participation in the registry. In addition, the amount of patients enrolled so far and allocated to a treatment approach are reported. RESULTS: According to the standard of treatment forms completed by 56 registered centers, fewer EVDs (Y1 55% Y2 46%) were used while more centers have implemented NEL (Y1 39%; Y2 52%) to treat nPHH. VAD (Y1 66%; Y2 66%) and VSGS (Y1 42%; Y2 41%) were used at a consistent rate during the 2 years. The majority of the centers used at least two different techniques to treat nPHH (43%), while 27% used only one technique, 21% used three, and 7% used even four different techniques. Patient data of 110 infants treated surgically between 9/2018 and 2/2021 (13% EVD, 15% VAD, 30% VSGS, and 43% NEL) were contributed by 29 centers. CONCLUSIONS: Our results emphasize the varying strategies used for the treatment of nPHH. The international TROPHY registry has entered into a phase of growing patient recruitment. Further evaluation will be performed and published according to the registry protocol.

[Downregulation of human CCR5 receptor gene expression using artificial microRNAs].

Chemokine receptor CCR5 is essential for human immunodeficiency virus (HIV) entry into the sensitive cells. The CCR5 inactivation is believed to be one of the promising approaches in HIV therapy, including gene therapy. A powerful mechanism that enables to regulate gene expression is RNA interference which could be exploited to knockdown CCR5 gene. Here, three artificial microRNAs directed against the human CCR5 receptor gene were generated and their silencing activity in indicator cells developed on the basis of the HT1080 cell line was evaluated. Multiplexing of two or more artificial microRNAs in one transcript has been demonstrated to enhance the gene silencing. A 95% reduction of the CCR5 expression has been achieved using the most efficient microRNA combination.

[Safety of retroviral vectors in gene therapy].

Retroviral vectors are widely used in gene therapy and found to be an effective tool for the delivery of genetic constructs into cells. A unique feature of these vectors is the ability to incorporate therapeutic genes into a chromosome that ensures its passage to all progeny cells and enables to cure the diseases requiring genetic correction of dividing cells such as hematopoietic cells or skin cells. Retroviral vectors have been successfully used in gene therapy clinical trials for the treatment of 2 forms of severe combined immunodeficiencies and some other hereditary blood disorders. However, the integration of the vector into the chromosome was accompanied by genotoxicity and caused development of hematologic malignancies in several patients. Later it was shown that genotoxicity is not a general feature of retroviral vectors but it depends on many factors. In the present article we discuss safety issues concerning the use of different retroviral vectors in gene therapy. The description of modern vectors which designed to avoid the genotoxicity and other possible side effects are given.

[Treatment of HIV-infection by means of gene therapy].

Current methods of HIV treatment can contain a progression of the disease; however they do not lead to a cure. Lifelong antiretroviral therapy is therefore necessary, leading to problems of cost and toxicity of chemical drugs. The recent advances in science have allowed a new approach to the HIV-treatment - gene therapy. In the present publication we focus on one strategy of the gene therapy called "intracellular immunization". The strategy is based on the introducing of antiviral genes into the HIV-sensitive cells. We highlight the mechanisms of action of various antiviral genetic agents and discuss some issues concerning target cells and genes delivery. Finally we summarize the results of certain gene therapy clinical trials.

[Progress in gene therapy].

Recent progress in gene therapy, current status of investigations in this area of experimental medicine are reviewed. Much attention is given to gene-therapeutic approaches the efficacy of which is proved in clinical trials.

[Protease and reverse transcriptase genetic polymorphism in HIV type 1 subtype A variants predominating in cis countries].

To define frequencies of drug resistance mutations among HIV-1 variants circulating within the territory of Russia, subtype A HIV-1 nucleotide sequences encoding protease and reverse transcriptase were analyzed. The analysis was carried out in 141 antiretroviral-naive individuals. Low frequency (less than 1%) of primary drug resistance mutations was shown. However, high frequencies of secondary mutations V77I in protease and A62V in RT (67% H 63%, respectively) linked to each other in most cases were observed. The HIV-1 isolates bearing both substitutions (MutV77I/A62V) were also characterized by the presence of several synonymous mutations, suggesting common origin for these viruses. HIV Biochip Hybridization microarray and/or Restriction fragment-length polymorphism analyses were performed to characterize gene pol polymorphism in additional 178 subtype A HIV-1 isolates. Among total 319 samples studied, Mutv77IA62V variant accounted for 56%, and was found to predominate in Russia in terms of both its geographical distribution and number of cases caused. Moreover, these viruses were prevalent in the regions known to have highest incidence of HIV-1 infection (Irkutsk, Samara, and Moscow regions). In addition, three other variants were found: viruses not containing the substitutions V77I or A62V, and variants bearing only one of them. Evolutional relationships between all four HIV-1 variants, as well as potential impact of the gene pol polymorphism on HIV-1 replicative fitness and drug resistance development are discussed.

[Comparative evaluation of the antitumor, cytogenetic and immunodepressive effects of dimetinur when used perorally and parenterally]. / Sravnitel'naia otsenka protivoopukholevogo, tsitogeneticheskogo i immunodepressivnogo éffektov dimetinura pri peroral'nom i parenteral'nom primenenii.

The dimethylnitrosourea action after oral and parenteral administration was comparatively evaluated on the basis of criteria for the antitumour and cytogenetic activity, as well as for the immune (T-cell) reactivity of tumour-bearing and intact animals. A considerable antitumour effect and the induction of the overload chromosomal aberrations in tumour cells with the complete preservation of bone marrow cells were observed during the oral drug application. Dimethyl nitrosourea-induced T-cell depression in murine spleen was transitory and reversible. Thus the oral administration of the drug was shown to be optimal for realization of its therapeutical activity with the least toxic side effect on the host normal hemopoietic and immunocompetent cells.

[Dynamics of the chromosomal damages to Ehrlich ascitic cancer and bone marrow cells of mice by the antitumor preparation dimetinur]. / Dinamika povrezhdenii khromosom kletok astsitnogo raka Erlikha i kostnogo mozga myshei protivoopukholevym preparatom dimetinur.

The dimetinur effect upon chromosomes of the Ehrlich ascite carcinoma and bone marrow cell populations was analyzed by the cytogenetic method for 10 days after i.p. and i. v. drug administration in doses from 50 to 150 mg/kg. Kinetic regularities of changes in a fraction of cells with chromosome breakages and in the number of broken chromosomes per cell as well as "dose-effect" relations are determined. A linear correlation is established between the level of residual chromosome damages in a population of tumour cells and the coefficient of activity chi* characterizing the antitumour effect of dimetinur. Selectivity of the dimetinur mutagenic action expressed as a more deep and prolonged damage of the genetic system of tumour cells as compared to bone marrow cells of tumour-bearing animals is shown under conditions of a pronounced therapeutic activity.

[A trial of the efficacy of using the synthetic antioxidant dibunol in the chemotherapy of experimental tumors]. / Opyt éffektivnosti primeneniia sinteticheskogo antioksidanta dibunola pri khimioterapii éksperimental'nykh opukholei.

We studied the therapeutic efficiency of the synthetic antioxidant Dibunol alone and in combination with cytostatic drugs on animals with experimental tumors after simultaneous and successive administration. Introduction of Dibunol to animals (tumor carriers) soon after the transplantation of tumor cells had adverse consequences, stimulated tumor growth and reduced the life span of animals. Dibunol exerted a therapeutic effect only in cases of developed tumors. Simultaneous introduction of cytostatic drugs (cyclophosphamide, dimethylnitrosourea, ADEKO) and Dibunol to tumor carriers was efficient and increased the mean life span of animals. The antitumor effect of the cyclophosphamide-Dibunol combination did not, in practice, depend on the interval between introductions of these drugs. However, the mean life span and survival of animals were somewhat higher the intervals between introductions or simultaneous application of the drugs were short.

[Antiretroviral therapy of acute HIV-infection]. / Antiretrovirusnaia terapiia ostroi VICh-infektsii.

AIM: To study efficacy of a short course of highly active antiretroviral therapy (HAART). MATERIAL AND METHODS: Acute HIV infection was verified in 16 infected subjected basing either on seroconversion (by immunoblotting) or a documented negative result 6-12 months before registration of a positive result for HIV antibodies. RESULTS: HAART was given to 12 patients (combivir + nevirapin). The number of CD+ lymphocytes (by median) 1 month after the treatment increased by 185 cells/mcl, 3 months after the treatment--by 215 cells/mcl. After 1-month therapy viral load (median) diminished by 2.02 log10 copy/ml, after 3 months--by 2.31 log10 copy/ml. 71% patients had HIV RNA under 400 copy/ml. Untreated patients showed changes neither in CD4+ lymphocytes number nor in viral load. The study continues. CONCLUSION: HAART is used at the stage of acute infection in the presence of psychic trauma provoked by establishment of HIV-infection diagnosis. Therefore, it is necessary to consult the patients for preparing them for treatment and to maintain compliance.

[Efficacy of cytoprotective agent Mexicor in urgent cardiology]. / éffektivnost' tsitoprotektora Meksikora v neotlozhnoi kardiologii.

AIM: To study efficacy of cytoprotector mexicor in patients with unstable angina (UA), acute myocardial infarction (MI), hypertensive crises (HC) in combined therapy with conventional drugs. MATERIAL AND METHODS: An open randomized study included 338 patients with acute forms of ischemic heart disease (IHD) and arterial hypertension running with crises. Combined therapy of 20 patients with UA, 90 patients with MI and 43 patients with HC (study groups) was supplemented with mexicor in a dose 6-9 mg/kg/day. The control matched patients (20, 86 and 79 patients, respectively) received conventional treatment alone. The effects of the treatments were assessed by ultrasound investigation of the heart in M-, B- and Doppler modes, by ECG and arterial pressure 24-h monitoring, by activity of lipid peroxidation (LPO). RESULTS: Adjuvant therapy of urgent cardiological conditions with mexicor diminished oxidant stress, left ventricular dysfunction. In MI patients mexicor promoted reduction of the akinesia zones, recovery of disturbed segmentary contractility. In UA patients mexicor contributed to more pronounced decrease in the frequency, duration and severity of myocardial ischemia, enhanced stabilization of angina. In HC patients mexicor promoted earlier normalization of a 24-h AP profile and variability of cardiac rhythm, recurrence rate of HC decreased 2-fold. CONCLUSION: The addition of mexicor to conventional therapy of UA, MI, HC improves clinical course of these diseases, reduces oxidant stress, accelerates recovery of cardiac contractility and left ventricular diastolic function, normalization of central hemodynamics.

[Determination of HIV RNA concentration for evaluation of the efficacy of combined therapy of HIV infection]. / Opredelenie kontsentratsii RNK VICh dlia otsenkii éffektivnosti kombinirovannoi terapii VICh-infektsii.

Plasma HIV RNA (viral load) and count of CD4+ T cells were evaluated in 23 patients with HIV infection treated with invirase (1800 mg/day), zidovudine (600 mg/day), and zalcitabine (2.25 mg/day) for 6 months in order to evaluate the efficiency of antiretroviral therapy. Viral load was measured by AMPLICOR HIV-1 Monitor test. The reproducibility of HIV RNA measurements was in line with reported data (CV 15-41%), allowing highly accurate (15%) evaluation of RNA in a standard control sample provided by National Institute for Biological Standards and Control, Great Britain. Plasma HIV RNA concentration decreased to an undetectable level (below 400 RNA copies/ml plasma) after 6 months of treatment in 52.2% patients. In 17.4% the therapy failed, and in 30.4% it resulted in a reduction of viral load to > 1 lg, although HIV RNA was still detected in the plasma after 6-month therapy. The count of CD4+ T cells increased by 9.5%. Changes in the viral load outstripped changes in CD4+ cells. Viral load was in high correlation with the count of CD4+ lymphocytes: -0.53, p = 0.01 before treatment and -0.61, p = 0.002 after 6-month treatment.