A New Chimeric Antibody against the HIV-1 Fusion Inhibitory Peptide MT-C34 with a High Affinity and Fc-Mediated Cellular Cytotoxicity.

Peptides from heptad repeat (HR1 and HR2) regions of gp41 are effective inhibitors of HIV-1 entry that block the fusion of viral and cellular membranes, but the generation of antibodies highly specific for these peptides is challenging. We have previously described a mouse hybridoma that recognizes MT-C34-related peptides derived from HR2. It was used for the selection of HIV-1-resistant CD4 lymphocytes engineered to express the MT-C34 peptide via a CRISPR/Cas9-mediated knock-in into the locus.

Photo- and X-ray Induced Cytotoxicity of CeF-YF-TbF Nanoparticle-Polyvinylpyrrolidone-"Radachlorin" Composites for Combined Photodynamic Therapy.

The CeYTbF nanoparticles with a CeF hexagonal structure were synthesized using the co-precipitation technique. The average nanoparticle diameter was 14 ± 1 nm. The luminescence decay curves of the CeYTbF nanoparticles (λ = 541 nm, D-F transition of Tb) conjugated with Radachlorin using polyvinylpyrrolidone coating as well as without Radachlorin were detected.

Evaluation of the Effectiveness of Bone Grafting in Alveolar Ridge Augmentation Using the Two-Stage Splitting Technique.

Stimulation of neoosteogenesis is the main mechanism of osseointegration during installation of dental implants, bone tissue recession, and alveolar process augmentation in adentia. In experiments on miniature pigs, we used the technology of two-stage splitting of the ridge of the alveolar process of the mandible in combination with a xenograft that was placed between the fragments of the split bone plate. The morphology of the reparative process and the distribution of osteogenic differentiation markers in the compact and trabecular bone of the alveolar crest were studied.

BMP-2 and IL-1β as Markers of Nasal Mucosa Inflammation in Rhinosinusitis with Nasal Polyps.

Rhinosinusitis with nasal polyps is characterized by chronic inflammation and hyperplasia of the nasal mucosa. The key mechanism for polyp formation is the expression of molecules that regulate proliferation and inflammation. We studied immunolocalization of bone morphogenetic protein-2 (BMP-2) and IL-1β in the nasal mucosa in patients aged 35-70 years (n=70, mean age 57.

Neurotoxic and cytoprotective mechanisms in the ischemic neocortex.

Neuronal damage in ischemic stroke occurs due to permanent imbalance between the metabolic needs of the brain and the ability of the blood-vascular system to maintain glucose delivery and adequate gas exchange. Oxidative stress and excitotoxicity trigger complex processes of neuroinflammation, necrosis, and apoptosis of both neurons and glial cells. This review summarizes data on the structural and chemical changes in the neocortex and main cytoprotective effects induced by focal ischemic stroke.

On-Bone Fixation of Free Gingival Graft Induces the Expression of Osteogenic Differentiation Markers in the Mandible of Miniature Pigs.

Purpose: To study the distribution of markers of mesenchymal stem cells (clusters of differentiation-29 and -44), osteoblasts (osteocalcin), and transforming growth factor-β1 in the mandibular tissue of miniature pigs after on-bone fixation of a free gingival graft.

Materials And Methods: Six outbred male miniature pigs 2 years of age were used in the study. In the external surface of the mandible, the periosteum was excised from all over the free gingival graft fixation area.

Topography of neurotrophins in the rat neocortex and their role in neuron apoptosis after experimental ischemic stroke.

Neuronal loss due to apoptosis after ischemic injury depends on the trophic support of neurons and cytoprotective effects of neurotrophins (NTs). Different NTs may activate both pro- and antiapoptotic factors. Their distribution in the ischemic core (IC) and penumbra (IP) has been poorly studied.

Localization of VEGF, TGF-β1, BMP-2, and Apoptosis Factors in Hypertrophic Nonunion of Human Tubular Bones.

We studied localization of VEGF, TGF-β1, BMP-2, caspase-3, Bcl-2, and TNFα in the callus samples obtained from 5 patients (4 women and 1 man) aged 41-53 years during planned surgery for nonunion and pseudarthrosis of the clavicle (n=1), ulna (n=1), femur (n=1), and tibia (n=2) bones. Two control groups included material of hypertrophied callus (n=3) with consolidated fractures of long bones and samples of intact bones (n=3) obtained by postmortem autopsy of subjects without pathology of the musculoskeletal system. A nonuniform distribution of the studied markers was revealed.

Engineering T-Cell Resistance to HIV-1 Infection via Knock-In of Peptides from the Heptad Repeat 2 Domain of gp41.

Previous studies suggest that short peptides from the heptad repeat 2 (HR2) domain of gp41 expressed on the cell surface are more potent inhibitors of HIV-1 entry than soluble analogs. However, their therapeutic potential has only been examined using lentiviral vectors. Here, we aimed to develop CRISPR/Cas9-based fusion inhibitory peptide knock-in (KI) technology for the generation and selection of HIV-1-resistant T cells.

HIV-1 and HTLV-1 Transmission Modes: Mechanisms and Importance for Virus Spread.

So far, only two retroviruses, human immunodeficiency virus (HIV) (type 1 and 2) and human T-cell lymphotropic virus type 1 (HTLV-1), have been recognized as pathogenic for humans. Both viruses mainly infect CD4+ T lymphocytes. HIV replication induces the apoptosis of CD4 lymphocytes, leading to the development of acquired immunodeficiency syndrome (AIDS).

On-Bone Fixation of Free Gingival Graft Induces an Osteoinductive Effect in Human Alveolar Bone.

We examined alveolar bone samples in the area of on-bone fixation of a free gingival graft performed during surgery in patients aged 37-55 years with a diagnosis of secondary partial adentia of the upper and lower jaws. Six months after fixation of the graft in the alveolar bone, foci of neoosteogenesis were found in the contact zone. They were characterized by the appearance of appositional lines, cords of basophilic osteoblasts, and growing osteons.

Dynamics of Renewal of Cell Populations of the Bone Tissue on the Surface of Titanium Implants with Bioactive Coating during Fracture Modeling in Rats.

Localization of PCNA, CD44, osteocalcin, Mdm2, p53, and caspase-3 on the surface of implant with calcium phosphate and hydroxyapatite coating was studied by immunocytochemical method in a model of femur fracture in rats. PCNA, Ost, CD44, and Mdm cells were found in the periosteum, in the layer of the outer surrounding plates, and in the connective tissue of the Haversian canals. Cell density increased on day 7 after fracture and then decreased by day 30.

[Combination of ARE and HRE cis- Regulatory Elements Elevates the Activity of Tumor-Specific hTERT Promoter].

Tumor-specific promoters and cis-regulatory genetic elements are used for transcriptional control of therapeutic transgene expression in cancer gene therapy. HRE (hypoxia response element) and ARE (anti-oxidant response elements) cis-regulatory elements are targets for HIF1 and Nrf2 transcriptional factors, respectively, and mediate activation of gene transcription in a response to hypoxia and oxidative stress, characteristic of most solid tumors. Due to these features HREs and AREs are used in genetic constructs for cancer gene therapy to provide tumor-specific therapeutic transgene expression or replication of oncolytic adenovi-ruses.

Osteogenic and Regenerative Potential of Free Gingival Graft.

We studied immunolocalization of CD29, CD44, osteocalcin, and TGF-β1 in the bone tissue of the mandible of miniature pigs with extra-bone fixation of a free gingival graft. Three months after surgery, neoosteogenesis foci with high expression of the studied markers were found in the contact area of the free gingival graft with the alveolar bone. The markers were localized in the layer of external circumferential lamellae, on the surface of concentric lamellae of the growing osteons, and in the connective tissue of the Haversian canals.

The effect of calcium phosphate biodegradable coatings of titanium implants on cell differentiation and apoptosis in rat bone tissue after experimental fracture.

Background: The effectiveness of bone repair is determined by the balance of proliferative and destructive factors in the fracture union site. It can be enhanced by using various nanostructured materials possessing osteoinductive properties, in particular titanium implants with biodegradable calcium phosphate coatings. The effects of these coatings on the state of stem cells, their differentiation and distribution in the repair zone is unknown.

Brain-Derived Neurotrophic Factor (BDNF) As a Regulator of Apoptosis under Conditions of Focal Experimental Stroke.

The immunolocalization of apoptotic factors in rat neocortex was studied on the model of permanent occlusion of the middle cerebral artery with administration of exogenous BDNF. We revealed heterogeneous distribution of pro- and anti-apoptotic factors in the stroke area and in the surrounding penumbra, where caspase-3 and p53 cells were found. Their number was maximum on day 3 of ischemia.

Study of anti-virus actions of metabolites of lactobacteria.

Objective: The aim: of the work was to study the antiviral activity of the metabolites of the probiotic strain Lactobacillus rhamnosus GG (LGG or ATCC 53103) regarding clinical strains of enteroviruses (Coxsackie B-5, ECNO21) isolated from the feces of intestinal infections.

Patients And Methods: Materials and methods: The object of the study was substrate-dependent cell cultures of HeLa, Vero, Hep-2 lines. The titer of the virus was determined by the presence of a clear cytopathic action (CPA) in the monolayer infected cells of the virus.

[Expression of pro- and anti-apoptotic molecules in the mucous membrane of the nasal cavity with polypous rhinosinusitis].

Objective: To study the expression of proapoptotic (p53, p21) and antiapoptotic (MDM2) factors, as well as the distribution of proliferating PCNA-immunoreactive cells in the nasal mucosa in various types of polyposis rhinosinusitis (PRS).

Material And Methods: We studied the immunolocalization of proapoptotic (p53, p21) and antiapoptotic (MDM2) factors, as well as the distribution of proliferating PCNA immunoreactive cells in the mucous membrane of the nasal cavity for various types of polypous rhinosinusitis.

Results And Discussion: Comparing with the control group, increased expression of all factors is detected.

Structural and chemical changes in glial cells in the rat neocortex induced by constant occlusion of the middle cerebral artery.

Stroke-induced changes in neuroglia determine the basic conditions for the survival and damage of neurons in the ischemic core. Here, we studied the immunolocalization of glial cell line-derived neurotrophic factor (GDNF), glial fibrillary acidic protein (GFAP), ionized calcium-binding adaptor molecule 1 (Iba-1), and S-100β in the rat parietal cortex after constant occlusion of the middle cerebral artery. These cytoplasmic proteins are specific for different glial cell types.

Xenobiotic Response Elements (XREs) from Human CYP1A1 Gene Enhance the hTERT Promoter Activity.

A hybrid 6XRE-hTERT promoter consisting of the hTERT tumor-specific promoter and six copies of the XRE element from the CYP1A1 human gene promoter was created. Using a human lung cancer cells as a model, we showed that XRE elements in the hybrid promoter greatly increase the activity of the hTERT promoter and ensure the reporter gene transcriptional activation in response to the treatment of the cells with the AhR ligand benzo(a)pyrene. However, similar effects were also observed in normal human bronchial epithelial cells HBEpC, which indicates the loss of the tumor-specific activity by the 6XRE-hTERT hybrid promoter.

[Human Antithrombin III Minigene with an Optimized Splicing Pattern].

Antithrombin III (AT3) belongs to the superfamily of serine protease inhibitors (serpins) and is a major anticoagulant in physiological conditions. Based on SERPINC1 gene, a minigene coding for human AT3, which is valuable for medicine and biotechnology, was constructed by minimizing the size of lengthy introns and preserving the splicing site-flanking sequences. An analysis of the minigene splicing pattern identified one correct AT3 transcript and two alternatively spliced transcripts, which formed either due to minigene exons 2 and 3 skipping or an aberrant exon insertion via splicing at cryptic splicing sites in intron 1 of the minigene.

Gaseous transmitters in human retinogenesis.

Endogenous gaseous transmitters (nitric oxide, carbon monoxide, and hydrogen sulphide) form a special neuromodulation system mediating the development and modification of nerve centers. Here, we examined the localization of key gaseous transmitter enzymes: cystathionine β-synthetase (CBS), cystathionine γ-lyase (CSE), heme oxygenase 2 (HO-2), and constitutive NO synthase (nNOS) in the fetal human retina at different stages of development. The number of CBS- and CSE-positive photoreceptors and intermediate retinal neurons was high in trimester I and gradually decreased to the end of trimester III.

The topography and proliferative activity of cells immunoreactive to various growth factors in rat femoral bone tissues after experimental fracture and implantation of titanium implants with bioactive biodegradable coatings.

Background: Biodegradable implant coatings promote proliferation and expression of BMP-2, VEGF, and TGF-β2 genes and enhance BMP-2, VEGF, and TGF-β2 regulatory effects at different stages of reparative osteogenesis.

Objective: To study the topography and ratio of PCNA-, VEGF-, BMP-2-, and TGF-β2-immunoreactive cells in rat femoral bone after closed fracture and implantation of titanium implants with biodegradable calcium phosphate and hydroxyapatite coatings.

Methods: Standard titanium implant screws and similar implants with bioactive coatings were used.

Production of Recombinant Proteins in the Milk of Transgenic Animals: Current State and Prospects.

The use of transgenic animals as bioreactors for the synthesis of the recombinant proteins secreted into milk is a current trend in the development of biotechnologies. Advances in genetic engineering, in particular the emergence of targeted genome editing technologies, have provided new opportunities and significantly improved efficiency in the generation of animals that produce recombinant proteins in milk, including economically important animals. Here, we present a retrospective review of technologies for generating transgenic animals, with emphasis on the creation of animals that produce recombinant proteins in milk.

[Insertion of Multiple Artificial Introns of Universal Design into cDNA during Minigene Construction Assures Correct Transgene Splicing].

The presence of introns is often required for efficient transgene expression. The use of full-length genes for transgenesis is associated with technical difficulties due to the large size of the genetic construct. To solve this problem, we recently suggested a universal design of small artificial introns that ensures efficient splicing.