HspBP1 in Complex with the Peptide of the Innate Immunity Protein Tag7 is Able to Lyse Tumor Cells Carrying TNFR1 Receptor.

The search for new cytotoxic agents capable of lysing tumor cells is an important task in the fight against cancer. Here we have shown that the HspBP1 protein, the chaperone of the heat shock protein Hsp70, is able to form a complex with the previously discovered peptide (17.1) of the innate immunity protein Tag7. Experiments using thermophoresis demonstrated that the affinity of the Tag7 protein peptide 17.1 to the HspBP1 molecule is 100 times higher than that of the full-sized Tag7 molecule. The addition of the 17.1-HspBP1 complex to tumor cells induces apoptosis and necroptosis in them. The results obtained in this work can be used to develop promising antitumor drugs.

Modular Nanotransporters Capable of Binding to SARS-CoV-2 Virus Nucleocapsid Protein in Target Cells.

On the basis of literature data, an antibody-like molecule, monobody, was selected that is capable of interacting with the nucleocapsid protein (N protein) of the SARS-CoV-2 virus with a high affinity (dissociation constant 6.7 nM). We have previously developed modular nanotransporters (MNTs) to deliver various molecules to a selected compartment of target cells. In this work, a monobody to the N protein of the SARS-CoV-2 virus was inserted in the MNT using genetic engineering methods. In this MNT, a site for the cleavage of the monobody from the MNT in endosomes was also inserted. It was shown by thermophoresis that the cleavage of this monobody from the MNT by the endosomal protease cathepsin B leads to a 12-fold increase in the affinity of the monobody for the N protein. Cellular thermal shift assay showed the ability of the obtained MNT to interact with the N protein in A431 cells transfected with the SARS-CoV-2 N protein fused to the mRuby3 fluorescent protein.

Intracellular Delivery of an Antibody-Like Molecule Capable of Inhibiting c-Myc.

A modular nanotransporter (MNT) carrying the sequence of an antibody-like molecule, anti-c-Myc nanobody, was synthesized and characterized. It was demonstrated that the created MNT is able to interact with the target protein, c-Myc oncogene, with a dissociation constant of 46 ± 14 nM, internalize into target cells, change Myc-dependent expression, and exert an antiproliferative effect.

Selection of an Amino Acid Site with One of the Fastest Cleavage Kinetics by the Endosomal Protease Cathepsin B for Potential Use in Drug Delivery Systems.

On the basis of known published data, six peptide sequences were selected that are potentially capable of being rapidly cleaved by the endosomal protease cathepsin B. For comparison, the cleavage of common linker sequences, polyglycine and polyglycine-serine, by cathepsin B was also studied. Different ends of these peptides were labeled with sulfoCyanine3 and sulfoCyanine5 fluorescent dyes, between which Förster resonant energy transfer (FRET) is possible. The kinetics of cleavage of peptides by cathepsin B was studied on a multimodal plate reader by FRET signal reduction. FKFL and FRRG cleavage sites have been shown to be the most suitable for potential use in various drug delivery systems. These sites are much more efficiently cleaved under slightly acidic conditions of endosomes than at neutral extracellular pH.

An unusual variant of the atlantomastoid muscle.

Knowledge of anatomical variations can be of use to clinicians and surgeons when, for example, viewing images of a patient or performing operations. Such knowledge can minimise the risk of iatrogenic complications. Herein, we present a case of a variant atlantomastoid muscle. The muscle was identified on the left side in an adult cadaver. The muscle's measurements and anatomical relationships are presented as well as a review of salient literature. We hope that increased knowledge of anatomical variants in the suboccipital region can improve patient care.

Unusual sensory innervation of the dorsal hand and why we should bear this variation in mind.

Detailed knowledge of the anatomy and different variations of the superficial branch of the radial nerve could be of great importance not only to anatomists but also to clinicians. A predominant radial nerve supply to the dorsum of the hand is rare. Herein, we present an unusual case of unilateral sensory innervation of the dorsal hand found during routine anatomical dissection of a 72-year-old at death male Caucasian cadaver. We also present a brief discussion of the reported variation and emphasize its potential clinical implications.

Duplication of the foramen rotundum: A rare case report.

Variations at the skull base can result in misinterpretation of radiological imaging and occasionally, iatrogenic injury. Here, we describe, to our knowledge, the second reported case of a duplicated foramen rotundum. The morphometrics of this finding are documented and the anatomy and potential clinical consequences of such an anatomical variation discussed. Such a finding is of archival value. Additionally, foramen rotundum duplication such as found in our case might also lead to complications while using, for example, transsphenoid approaches to the middle cranial fossa as well as various transfacial treatments for trigeminal neuralgia which rely on observing the foramina around the foramen ovale on fluoroscopy for correct positioning of needles and catheters.

Previously unreported variant of the rectus femoris muscle.

Although uncommon, variant muscular variations can occur related to the quadriceps femoris. Such variations might be encountered in the clinical setting so these should be documented. Here, we report three additional heads related to the rectus femoris muscle identified during routine dissection of the right thigh. To our knowledge, such a variation has not been previously reported.

A unique variation of a four-bellied digastric muscle named "real quadrigastric muscle": a case report and literature review.

From a topographical standpoint, the digastric muscle is key to the formation of several triangles of the neck, which are of the utmost clinical significance. Herein, we present a previously unrecognised variation of the digastric muscle: a quadrigastric muscle with two accessory bellies originating from the body and angle of the mandible and inserting to the intermediate tendon. Three new triangles are demarcated between the four bellies of the aberrant muscle. Detailed knowledge of variations of the digastric muscle, changing the borders and relationships of the topographic triangles, is paramount for radiologists and surgeons operating on the anterior region of the neck.

Variant origin of three main coronary ostia from the right sinus of Valsalva: report of a rare case.

Observing anomalies in the origin of the coronary arteries is a rare but recognised scenario during coronarography. All the major coronary arteries originating from the right sinus of Valsalva is an extremely rare anomaly, its reported incidence being 0.008% in angiographic studies. Most coronary artery variations are benign and are therefore found accidentally or postmortem. However, some anomalies in the origin of the coronary arteries are associated with myocardial ischaemia and a higher risk of sudden cardiac death. Herein, we report a sporadic case of anomalous origin of the coronary arteries, in which the right coronary artery, anterior interventricular artery and left circumflex artery arise separately from the right sinus of Valsalva, each originating from a separate ostium. Regardless of their low incidence rate, coronary artery anomalies can cause serious technical challenges during coronary angiography and percutaneous interventions because of the unusual location and course of the artery. Echocardiography, computed tomography, and magnetic resonance imaging can be useful in such cases.

Quantitative Description of the N-Protein of the SARS-CoV-2 Virus Degradation in Cells Stably Expressing It under the Influence of New Modular Nanotransporters.

Two eukaryotic cell lines, A549 and A431, with stable expression of the nucleocapsid protein (N-protein) of the SARS-CoV-2 virus fused with the red fluorescent protein mRuby3 were obtained. Using microscopy, the volumes of the cytoplasm and nucleus were determined for these cells. Using quantitative immunoblotting techniques, the concentrations of the N-mRuby3 fusion protein in their cytoplasm were assessed. They were 19 and 9 µM for A549 and A431 cells, respectively. Using these concentrations, the initial rate of N-protein degradation in the studied cells was estimated from the decrease in cell fluorescence. In A549 and A431 cells, it was the same (84 nM per hour). The approach of quantitatively describing the degradation process can be applied to analyze the effectiveness of a wide class of antiviral drugs that cause degradation of viral proteins.

Modular Nanotransporters Capable of Causing Intracellular Degradation of the N-Protein of the SARS-CoV-2 Virus in A549 Cells with Temporary Expression of This Protein Fused with a Fluorescent Protein mRuby3.

Modular nanotransporters (MNTs) containing an antibody-like molecule, monobody, to the N­protein of the SARS-CoV-2 virus, as well as an amino acid sequence that recruits the Keap1 E3 ligase (E3BP) were created. This MNT also included a site for cleavage of the E3BP monobody from the MNT in acidic endocytic compartments. It was shown that this cleavage by the endosomal protease cathepsin B leads to a 2.7-fold increase in the affinity of the E3BP monobody for the N-protein. Using A549 cells with transient expression of the N-protein fused with the fluorescent protein mRuby3, it was shown that incubation with MNT leads to a significant decrease in mRuby3 fluorescence. It is assumed that the developed MNTs can serve as a basis for the creation of new antiviral drugs against the SARS-CoV-2 virus.

Tag7-Mts1 Complex Activates Chemotaxis of Regulatory T Cells.

One of the basic features of immune system is the ability to sustain balance between activation and suppression of effector lymphocytes. In this process a key role belongs to the subpopulation of cells called regulatory T cells (Treg). Many cancer and autoimmune diseases are caused by malfunctions of Treg, and investigation of this subpopulation is important for development of new therapeutic approaches. In this study, we demonstrate that regulatory T cells can migrate along the concentration gradient of Tag7-Mts1 complex, and also they produce agents that induce blood cells migration.

Delivery of Antibody-Like Molecules, Monobodies, Capable of Binding with SARS-CoV-2 Virus Nucleocapsid Protein, into Target Cells.

Based on previous studies, two antibody-like molecules, monobodies, capable of high-affinity interaction with the SARS-CoV-2 nucleocapsid protein (dissociation constant of tens of nM) were selected. For delivery to target cells, genetically engineered constructs containing monobody and TAT peptide, placed either at the N- or C-terminus of the resulting polypeptide, were produced and expressed in E. coli. The construct with the highest affinity to the SARS-CoV-2 nucleocapsid protein was revealed with the use of thermophoresis technique. Cellular thermal shift assay demonstrated the ability of this construct to interact with the nucleocapsid protein within HEK293T cells transfected with the SARS-CoV-2 nucleocapsid protein fused to the mRuby3 fluorescent protein. Replacement of TAT peptide to S10 shuttle peptide, containing endosomolytic peptide, significantly improved the penetration of the construct into the target cells.

Nuclear moments of indium isotopes reveal abrupt change at magic number 82.

In spite of the high-density and strongly correlated nature of the atomic nucleus, experimental and theoretical evidence suggests that around particular 'magic' numbers of nucleons, nuclear properties are governed by a single unpaired nucleon1,2. A microscopic understanding of the extent of this behaviour and its evolution in neutron-rich nuclei remains an open question in nuclear physics3-5. The indium isotopes are considered a textbook example of this phenomenon6, in which the constancy of their electromagnetic properties indicated that a single unpaired proton hole can provide the identity of a complex many-nucleon system6,7. Here we present precision laser spectroscopy measurements performed to investigate the validity of this simple single-particle picture. Observation of an abrupt change in the dipole moment at N = 82 indicates that, whereas the single-particle picture indeed dominates at neutron magic number N = 82 (refs. 2,8), it does not for previously studied isotopes. To investigate the microscopic origin of these observations, our work provides a combined effort with developments in two complementary nuclear many-body methods: ab initio valence-space in-medium similarity renormalization group and density functional theory (DFT). We find that the inclusion of time-symmetry-breaking mean fields is essential for a correct description of nuclear magnetic properties, which were previously poorly constrained. These experimental and theoretical findings are key to understanding how seemingly simple single-particle phenomena naturally emerge from complex interactions among protons and neutrons.

Variations in the origin of the superior laryngeal artery and their clinical significance: a case report with a proposal for a new classification / Variaciones en el origen de la arteria laríngea superior y su significado clínico: reporte de un caso con una nueva propuesta de clasificación

SUMMARY: The superior laryngeal artery is the primary vessel providing the blood supply to the larynx. Commonly, it is derived from the superior thyroid artery. Different variations in the origin have been described in the current literature; knowledge of such variations is crucial for various surgical interventions of the larynx and surgical procedures in the lateral region of the neck regarding the carotid triangle. It should be noted that radiological studies, such as selective angiography of the thyroid gland, can also be misleading in cases of variations. Herein, we describe a case of bilateral superior laryngeal artery originating directly from the external carotid artery of the neck. The arteries at first have a transverse course and then pierce through the thyrohyoid membrane alongside internal laryngeal nerves. Moreover, we also review the known variations in the origin of the superior laryngeal artery and propose a new classification of all known variations.

RESUMEN: La arteria laríngea superior es el vaso principal que proporciona el suministro de sangre a la laringe. Comúnmente, se deriva de la arteria tiroidea superior. Han sido descritas diferentes variaciones en su origen y el conocimiento de éstas resulta crucial para las intervenciones quirúrgicas realizadas en la laringe, como también en los procedimientos quirúrgicos que se llevan a cabo en la región lateral del cuello, respecto al triángulo carotídeo. Cabe señalar que los estudios radiológicos, como la angiografía selectiva de la glándula tiroides, también pueden ser engañosos en casos de variaciones anatómicas. Aquí, describimos un caso de arteria laríngea superior bilateral que se originaba directamente de la arteria carótida externa. Las arterias al inicio tenían un curso transversal y luego atravezaban la membrana tirohioidea junto con los nervios laríngeos internos. Revisamos también las variaciones conocidas en el origen de la arteria laríngea superior y proponemos una nueva clasificación de todas las variaciones conocidas.

Among Antibody-Like Molecules, Monobodies, Able to Interact with Nucleocapsid Protein of SARS-CoV Virus, There Are Monobodies with High Affinity to Nucleocapsid Protein of SARS-CoV-2 Virus.

Seven amino acid sequences of antibody mimetics molecules, monobodies, capable of interacting with the nucleocapsid protein of the SARS-CoV virus, were taken from the literature. Nucleotide sequences of monobody genes were obtained by gene synthesis, which were expressed in E. coli and isolated using Ni-NTA chromatography. It was shown by thermophoresis that three of the seven selected antibody-like molecules can interact with high affinity (dissociation constant of tens of nM) with the nucleocapsid protein of the SARS-CoV-2 virus. For the remaining four monobodies, only low affinity binding with a dissociation constant of several µM was found.