In Vivo DYSF Gene Viral Delivery Provides a Histoprotective Effect in Skeletal Muscle Tissue in Dysferlin-Deficient Mice.

We studied the effects of a dual-vector DYSF gene delivery system based on adeno-associated virus serotype 9 capsids on pathological manifestations of dysferlinopathy in skeletal muscles of Bla/J mice lacking DYSF expression. The mice received intravenous injection of 3×1013 genomic copies of the virus containing the dual-vector system. M. gastrocnemius, m. psoas major, m. vastus lateralis, and m. gluteus superficialis were isolated for histological examination in 3, 6, and 12 weeks after treatment. Healthy wild-type (C57BL/6) mice served as positive control and were sacrificed 3 weeks after injection of 150 µl of 0.9% NaCl into the caudal vein. To detect dysferlin in muscle cryosections, immunohistochemical analysis with diagnostic antibodies was performed; paraffin sections were stained with hematoxylin and eosin for morphometric analysis. After administration of gene-therapeutic constructs, muscle fibers with membrane or cytoplasmic dysferlin location were detected in all examined muscles. The proportion of necrotic muscle fibers decreased, the number of muscle fibers with central location of the nucleus increased, and the mean cross-section area of the muscle fibers decreased.

[Microbiome of the external auditory canal and preoperative preparation of patients in otosurgery (literature review)]. / Mikrobiom naruzhnogo slukhovogo prokhoda i predoperatsionnaya podgotovka patsientov v otokhirurgii (obzor literatury).

OBJECTIVE: To analyze the effectiveness of methods of antiseptic and hygienic preoperative preparation of the operating field in otosurgery. MATERIAL AND METHODS: A review and an expert assessment of the corresponding research objectives of scientific publications contained in the PubMed and Google Scholar databases have been carried out. RESULTS: Various studies indicate a lower ototoxicity of 5-10% of the drug povidone-iodine compared to others. Drugs, such as Miramistin, Okomistin, Dioxidine, are actively used in otiatrics in Russia, but no data on their ototoxicity have been noted. The collected data point to the relevance and prospects of studying the microbiome of the external auditory canal, assessing its effect on the course of the postoperative period, depending on various methods of preparation (antiseptic, removal of earwax and hair depilation) of the surgical field for otosurgery.

[The main steps in treatment of the children with otitis media with effusion]. / Osnovy okazaniya meditsinskoi pomoshchi pri ekssudativnom srednem otite.

THE AIM: Of the investigation was to establish the standard and improve the treatment of otitis media with effusion (OME) in children. 361 children at age from 11 months to 18 years were inspected after tympanostomy during 2013-2018 years. The main diagnostic methods were: otoscopy, tympanometry, endoscopy, CT. MATERIAL AND METHODS: Treatment takes into consideration the reveal of OME: surgical initially. The tympanostomy preferable place is anterior-inferior quadrant. RESULTS: In cases with cleft palate or reccurence OME long-term tubes and balloonisation of ET are preferable. CONCLUSION: Authors received normalization of the hearing thresholds in 97.6% cases, but after surgery the patients have to be followed-up during 12-24 months.

[Obliteration of the middle ear cavity in children: the current state of the problem and our first experience]. / Obliteratsiya polostei srednego ukha u detei: sovremennoe sostoyanie problemy i nash pervyi opyt.

Chronic otitis media is one of the most common ear pathologies among children. This article provides a short overview of publications on surgical techniques. Currently, the most effective treatment is the surgical sanitation of the middle ear cavities. There are two fundamentally different approaches of operations in relation to the posterior wall of the external auditory canal - «open¼ and «closed¼ types. Both methods have their advantages and disadvantages, which are especially noticeable in pediatric revision otosurgery. Surgical obliteration of the sanitized middle ear cavities is an effective compromise option. However, the techniques of bone obliteration, which are actively applied among adults, are difficult if we discuss children ear. A way out of this situation may be the use of modern osteoplastic materials for filling large trepanation cavities. The article describes in detail the technique of revision sanitizing surgery on the middle ear using osteoplastic granules. Purpose of the study. The analysis of the first results of separate obliteration of paratympanic cavities in children using osteoplastic materials. MATERIAL AND METHODS: In the period from May 2018 to November 2020, on the basis of the Federal State Autonomous Institution «National Medical Research Center of Children's Health¼ of the Ministry of Health of Russia, 28 children aged from 6 to 17-year-old with chronic otitis media and cholesteatoma were operated using osteoplastic obliteration, who had previously sanitizing operation on the middle ear. All children on admission to the hospital and 6-12 months after the operation underwent a complex examination. The condition of each patient was assessed using the OMO-22 quality of life questionnaire, to which the parents answered the questions twice - before and 1 year after surgery. RESULTS: During the postoperative period none of the patients had clinical signs of recurrence of cholesteatoma, which was confirmed by CT scan. When assessing the quality of life of children using a questionnaire, the average score before the operation was 130.2±27.7, after the operation - 61.5±21.1. The indicator of the bone-air interval before surgery in patients averaged 29.8±9.7 dB, 1 year after surgery - 13.0±10.9 dB (p>0.05). FINDINGS: The first experience of using osteoplastic materials for obliterating the paratympanic spaces in children has shown high efficiency, ease of use and safety.

[Treatment of exudative (secretory) otitis media in young children taking into account the anatomical features of the auditory tube]. / Lechenie ékssudativnogo srednego otita u detei rannego vozrasta s uchetom anatomicheskikh osobennostei slukhovoi truby.

AIM: To study the efficacy and safety of treatment of exudative (secretory) otitis media (EOM) in children of early age taking into account the anatomical features of the auditory tube. MATERIAL AND METHODS: We examined and treated 30 children (51 ears) by balloon dilatation (BD) at the age of 1 to 3 years, with recurrent EOM (more than 6 months). The 21st child was diagnosed with a 2-sided process, and 9 with a one-sided process. All patients have previously unsuccessfully used conservative methods of treatment. According to the results of the examination and treatment, the children were divided into 2 groups. In group 1 (16 children, 30 ears) included children suffering from ESR in combination with hypertrophy of adenoids 2-3 art. with a block of pharyngeal mouths of auditory tubes, which at stage 1 of treatment was carried out endoscopic adenotomy and shunting of the tympanic cavities. In follow-up, this group of children after shunt loss was diagnosed with relapse of EOM (type B was registered on tympanometry). In this group, balloon dilation of auditory tubes was performed in the second stage after adenotomy and bypass surgery. In group 2 (14 children, 21 ear) included children with relapsing course of EOM, the adenoid vegetation 1 degree without block the pharyngeal orifices of the auditory tubes, which in the first phase was conducted on bypass the tympanic cavity. After the loss of shunts, during the control examination, a relapse of EOM was diagnosed, in connection with which, the children underwent balloon dilation of the auditory tubes. RESULTS: During tympanometry in 1 month. complete restoration of middle ear function in 17 children (56.7%, 28 ears) (tympanograms type A) was revealed. 4 children (13.3%, 6 ears) were diagnosed with partial recovery of middle ear function (tympanograms type C). 9 children (30%, 17 ears) retained exudate in the tympanic cavity, these patients underwent a second course of conservative therapy, after which the function of the middle ear was restored. 9 children after BD with persistent exudate in the middle ear cavities underwent tympanostomy (in this case, long-term wearing shunts were used). CONCLUSION: Balloon dilation of the cartilaginous part of the auditory tube is minimally invasive, effective in 70% of cases and a safe approach to the treatment of children suffering from a recurrent course of EOM.

[Surgical treatment of chronic otitis media in children using three-dimensional imaging]. / Khirurgicheskoe lechenie khronicheskogo srednego otita u detei s ispol'zovaniem trekhmernoi vizualizatsii.

The introduction of three-dimensional exoscopy has opened a new stage in microsurgery in recent years. Researchers from various specialties give mostly positive reviews. There are very few works on the use of three-dimensional imaging in otorhinolaryngology, and there are practically no such works on otosurgery. This article presents a short review of publications and our own clinical examples, which are the first detailed description of VITOM 3D KARL STORZ exoscope use for cholesteatoma surgery in children, published in the available world literature. The first experience allowed us to draw preliminary conclusions about the comparability of this method of intraoperative imaging with a microscope. In addition, we have identified additional benefits of the exoscope. Further research will make it possible to better adapt the technique to the peculiarities of otosurgery, and it should take its place in the arsenal of ENT surgeons, along with microscopy.

[Recist 1.1 Criteria in the Assessment of Lung Tumors].

The paper considers the main provisions of the 2009 RECIST guidelines (version 1.1.) criteria for a standard approach to measuring lung tumor lesions and assessing the dynamics of the tumor process during treatment. This paper contains a list of conditions for carrying out multislice spiral computed tomography, under which the latter becomes maximally reproducible for the same patient, as well as an assessment of the course of the disease does optimal.

[Ribonucleolytic activity of mycoplasmas].

Mycoplasmas are incapable of de novo synthesis of nucleotides and must therefore secrete nucleases in order to replenish the pool of nucleic acid precursors. The nucleolytic activity of mycoplasmas is an important factor in their pathogenicity. Bacterial ribonucleases (RNases) may produce a broad spectrum of biological effects, including antiviral and antitumor activity. Mycoplasma RNases are therefore of interest. In the present work, capacity of Acholeplasma laidlawii and Mycoplasma hominis for RNase synthesis and secretion was studied. During the stationary growth phase, these organisms were found to synthesize Mg(2+)-dependent RNases, with their highest activity detected outside the cells. Localization of A. laidlawii RNases was determined: almost 90% of the RNase activity was found to be associated with the membrane vesicles. Bioinformational analysis revealed homology between the nucleotide sequences of 14 Bacillus subtilis genes encoding the products with RNase activity and the genes of the mycoplasmas under study. Amino acid sequences of 4 A. laidlawii proteins with ribonuclease activity and the Bsn RNase was also established.

[B-cell lymphoma in a woman with chronic hepatitis C and mixed cryoglobulinemia of type 2].

A report is presented of a rare case of B-cell non-Hodgkin's lymphoma and severe exacerbation of cryoglobulinic vasculitis which developed in a female patient with chronic hepatitis C four years after achievement of a persistent virusological response to antivirus treatment. Causes of a specific course of the disease and development of the tumor in the absence of HCV in blood serum are discussed: latent HCV-infection in immune cells with persistent antigenic stimulation of B-lymphocytes, possibility of HCV-independent lymphoproliferation.

[Prognosis factors in imatinib mesilate therapy in patients with a chronic phase of Ph-positive chronic myeloid leukemia: data from a multicenter non-randomized trial in Russia].

AIM: To reveal prognostically significant factors affecting efficacy of glivek therapy in untreated (duration of the disease < or = 6 months) and pretreated (duration of the disease > 6 months) patients with chronic myeloid leukemia (CML) in a chronic phase. MATERIAL AND METHODS: A total of 338 patients (64 untreated and 274 pretreated) with a chronic-phase CML on glivek therapy entered the trial. RESULTS: Five-year survival on glivek was high (89, 98 and 88% in untreated and pretreated patients, respectively). Incidence of transformation in the acceleration phase and blast crisis was low both in untreated and pretreated patients (1.6 and 11%, respectively) and correlated with the rate of a complete cytogenetic response (CCR). Untreated patients had no factors affecting treatment efficacy negatively, CCR probability was 96%. Blastemia, thrombocytosis and splenomegaly reduced CCR probability significantly in pretreated patients. Slow reduction of the tumor mass, late achievement of a complete hematological response and a cytogenetic response decreased probability of CCR. CONCLUSION: Glivek is a drug of choice for patients with chronic-phase CML. High probability of CCR both in untreated and pretreated patients lowers the risk of the disease transformation into the phase of acceleration/blast crisis and raises overall survival in both groups.

[Adaptive reactions of mycoplasmas in vitro: "viable but unculturable forms" and nanocells of Acholeplasma laidlawii].

The adaptation of Acholeplasma laidlawii to conditions unfavorable for growth has been found to be accompanied by cell transformation into special morphological structures known as ultramicroforms (nanocells). The ratio of the cells of the two morphological types in the population depended on the growth conditions. Nanocells retained viability for a long time under conditions unfavorable for growth and showed resistance to stressors. Reduction in the cell size occurred due to unequal division, which involved the loss of cytoplasmic material. A. laidlawii ultramicroforms (nanocells) were able to restore proliferative activity and to revert to their initial vegetative form; they measured less than 0.2 microm and are the smallest cells known at present. Nanocells formed in vitro under exposure to abiogenic stressors may correspond to the A. laidlawii minibodies observed in infected plants upon exposure to biogenic stressors. The transformation of A. laidlawii cells into ultramicroforms was accompanied by condensation of the nucleoid, a change in the polypeptide spectrum, and a change in the availability of rRNA operons for in vitro amplification. All these changes are indicative of reorganization of the genetic and metabolic systems of mycoplasmas.

Rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography.

High-performance liquid chromatography (HPLC) is a recently introduced high-capacity automated method for detecting unknown mutations (denaturing HPLC) or for sizing DNA fragments under nondenaturing conditions. We have adapted the HPLC method for detection of loss of heterozygosity (LOH) and used glial tumours as a model to evaluate its sensitivity and specificity in comparison to conventional denaturing polyacrylamide gel electrophoresis. A total of 20 oligodendrogliomas (grades II and III), and five astrocytic tumours (grades III and IV) were analysed using 14 polymorphic microsatellite markers mapping to regions on chromosomes 1p, 19q, and 10q using both DNA-HPLC and denaturing gel electrophoresis. This study demonstrated complete concordance between both methods. However, unlike gel electrophoresis, HPLC is automated, does not require post-PCR processing, and does not require hazardous radioactive or expensive fluorescent labelling. Our data suggest that HPLC is a reliable and sensitive method for detection of allelic losses in tumour samples and it is a favourable alternative for high-sensitivity LOH detection in both research and diagnostic environments.

Molecular characterization of the breakpoint region associated with a constitutional t(2;15)(q34;q26) in a patient with multiple myeloma.

The molecular cloning of the translocation breakpoints from constitutional chromosome rearrangements in patients with a variety of human diseases has consistently led to the isolation of genes important in the development of the phenotype. We used fluorescence in situ hybridization (FISH) to analyze the breakpoint region of a constitutional chromosome translocation involving regions 2q34 and 15q26 observed in a patient with multiple myeloma (MM), a malignant disorder of plasma cells secreting monoclonal immunoglobulin. FISH analysis of this rearrangement showed that the chromosome 2-specific yeast artificial chromosome (YAC) 914E7 and the chromosome 15-specific YAC 757H6 span the translocation breakpoints, respectively. In order to characterize the location of the breakpoints further, somatic cell hybrids were constructed between mouse NIH3T3 cells and t(2;15)-bearing lymphoblastoid cells. Using these somatic cell hybrids, we have shown that the breakpoint on chromosome 2 lies between D2S3007 and D2S3004 and the chromosome 15 breakpoint lies between D15S107 and WI5967 (D15S836). YAC fragmentation has been used to define a 350 kb region containing the 15q26 breakpoint.

A novel member of the WD-repeat gene family, WDR11, maps to the 10q26 region and is disrupted by a chromosome translocation in human glioblastoma cells.

Allelic deletions of 10q25-26 and 19q13.3-13.4 are the most common genetic alterations in glial tumors. We have identified a balanced t(10;19) reciprocal translocation in the A172 glioblastoma cell line which involves both critical regions on chromosomes 10 and 19. In addition, loss of an entire copy of chromosome 10 has occurred in this cell line suggesting that the translocation event may provide a highly specific critical inactivating event in a gene responsible for tumorigenesis. Positional cloning of this translocation breakpoint resulted in the identification of a novel chromosome 10 gene, WDR11, which is a member of the WD-repeat gene family. The WDR11 gene is ubiquitously expressed, including normal brain and glial tumors. WDR11 is composed of 29 exons distributed over 58 kilobases and oriented towards the telomere. The translocation resulted in deletion of exon 5 and consequently fusion of intron 4 of WDR11 to the 3' untranslated region of a novel member, ZNF320, of the Krüppel-like zinc finger gene family. Since ZNF320 is oriented toward the centromere of chromosome 19, both genes appeared on the same derivative chromosome der(10). The chimeric transcript encodes the WDR11 polypeptide, which is truncated after the second of six WD-repeats. ZNF320 is also expressed in A172 cells, although it is not clear if the translocation affects the expression of the altered gene because of the presence of another unrearranged gene on chromosome 19. We suggest that, because of its localization in a region frequently showing LOH and the observation of inactivation of this gene in glioblastoma cells, WDR11 is a candidate gene for the frequently proposed tumor suppressor gene in 10q25-26 which is involved in tumorigenesis of glial and other tumors showing frequent alterations in the distal 10q region.

Identification of the promoter, genomic structure, and mouse ortholog of LGI1.

The human LGI1 gene is a leucine-rich, repeat-containing gene that was cloned from the t(10:19) breakpoint of the T98G glioblastoma cell line. The LGI1 gene maps to 10q24, a region of peak LOH in malignant gliomas, and is inactivated during the transition from low to high-grade brain tumors. Here we report detailed studies of the genomic structure of the LGI1 gene and its promoter. We have also cloned and characterized the mouse lgil gene, which is 97% homologous to the human gene at the amino acid level and 91% homologous at the nucleotide level. LGI1 contains 8 exons, where each of the four leucine-rich repeat units is contained in an individual 72-bp exon. The cysteine-rich regions flanking the LRR and the single trans-membrane domain do not occupy individual exons. Approximately 5-kb of the genomic region 5' to LGI1 was sequenced, but conventional CAAT and TATA motifs were not present within this sequence. A 597-bp fragment of this 5' sequence was cloned upstream of a promoterless luciferase gene and was shown to be sufficient to drive transcription. SSCP analysis of the coding region of LGI1 in 20 glioblastomas and five cell lines did not reveal any mutations. Because LGI1 expression is considerably downregulated in gliomas, we also investigated whether this was owing to changes in the methylation status of the promoter. Southern blot analysis and 5-azacytidine treatment did not show any appreciable difference in methylation status between normal brain and glioblastomas.

Radiation-induced meningioma: a distinct molecular genetic pattern?

Radiation-induced meningiomas arise after low-dose irradiation treatment of certain medical conditions and are recognized as clinically separate from sporadic meningioma. These tumors are often aggressive or malignant, they are likely to be multiple, and they have a high recurrence rate following treatment compared with sporadic meningiomas. To understand the molecular mechanism by which radiation-induced meningioma (RIM) arise, we compared genetic changes in 7 RIM and 8 sporadic meningioma (SM) samples. The presence of mutations in the 17 exons of the neurofibromatosis type 2 (NF2) gene, which has been shown to be inactivated in sporadic meningiomas, was analyzed in RIM and SM using single-strand conformation polymorphism (SSCP) and DNA sequencing. In contrast to SM, which showed NF2 mutations in 50% of specimens, no mutations were found in RIM. In addition, Western blot analysis of schwannomin/merlin protein, the NF2 gene product, demonstrated protein levels comparable to normal brain in 4/4 RIM tumor samples analyzed. Loss of heterozygosity (LOH) of genomic regions, which were reported for SM, was also analyzed in all cases of RIM using 22 polymorphic DNA markers. Allele losses were found on chromosomes 1p (4/7), 9p (2/7), 19q (2/7), 22q (2/7), and 18q (1/7). From these observations we conclude that unlike sporadic meningiomas, NF2 gene inactivation and chromosome 22q deletions are far less frequent in RIM, and their role in meningioma development following low dose irradiation is less significant. Other chromosomal lesions, especially loss of 1p, possibly induced by irradiation, may be more important in the development of these tumors.

[Interaction of oxytocin, laser and electromagnetic radiation on the persistence properties of Staphylococcus aureus]. / Vzaimodeistvie oksitotsina, lazernogo i élektromagnitnogo izulucheniia na persistentnye svoistva Staphylococcus aureus.

The suppressive action of oxytocin, heliumneon radiation and ultrahigh-frequency electromagnetic waves (UHF-therapy) on the persistence properties of S. aureus has been experimentally established. The effectiveness of the therapeutic actions under study in the treatment of patients with the prognosticated unfavorable course of purulent inflammatory diseases of soft tissues has been shown.

[Biological importance of the anti-carnosine activity of bacteria]. / Biologicheskoe znachenie antikarnozinovoi aktivnosti bakterii.

The anticarnosine activity has been found to be widely spread among bacteria, the degree of its manifestation depending on the source of isolation. The fact that anticarnosine activity plays a certain role in the phenomenon of persistence has been proved on the model of experimental staphylococcal infection in rats.