Effect of sodium alginate block type on the physicochemical properties and curcumin release behavior of quaternized chitosan-oxidized sodium alginate Schiff base hydrogels.

Controlling bioactive ingredients release by modulating the 3D network structure of cross-linked hydrogels is important for functional food development. Hereby, oxidized sodium alginate (OSA) with varying aldehyde contents was formed by periodate oxidation of sodium alginate (SA) with different ß-d-mannuronic acid (M) and α-l-guluronic acid (G) ratios (M/G = 1:2, 1:1, and 2:1) and its structure was characterized. Moreover, hydrogels were prepared via Schiff base and electrostatic interactions between quaternized chitosan (QCS) and OSA. The properties of hydrogels such as microstructure, thermal stability, swelling and controlled release were investigated. The results showed that OSA with M/G = 1:2 had the highest content of aldehyde groups, and the hydrogel formed by it and QCS had higher thermal stability and a denser network structure with the lowest equilibrium swelling rate, which could better control the release of curcumin. Additionally, it had good self-healing and can recover rapidly after the rupture of its network structure.

Low-symmetry A3 B-type 6H-1,4-diazepinoporphyrazines with anti-Kasha effect as promising photosensitizers.

A series of tribenzo[g,l,q]-6H-1,4-diazepino[2,3-b]porphyrazines has been synthesized. A temperature-dependent steric effect was applied in the mixed Linstead macrocyclization of phthalonitrile and 5,7-bis(2'-arylethenyl)-6-propyl-6H-1,4-diazepine-2,3-dicarbonitrile to achieve high yield of low-symmetry A3 B-type Mg(II) tribenzo[g,l,q]-6H-1,4-diazepino[2,3-b]porphyrazinate. The analysis of photophysical and photochemical properties of the obtained complexes showed the anti-Kasha effect: the ultrafast spin changes successfully compete with the IC. TD-DFT calculations showed that the presence of 1,4-diazepine heterocycle in the porphyrazine structure leads to the formation of additional charge-transfer triplet state T2 . We propose, it could participate in the pumping of T1x state alongside with T1y state (these states are degenerate in D4h symmetry) and, therefore, increase singlet oxygen (1 Δg ) generation. Stable micellar nanoparticles have been obtained based on the tribenzo[g,l,q]-6H-1,4-diazepino[2,3-b]porphyrazine Mg(II) and Zn(II) complexes using polyvinylpyrrolidone. The nanoparticles effectively interact with model biological structures (FBS and brain homogenate), leading to disaggregation of the macrocycles. They also exhibit pronounced phototoxic effects in MCF-7 cells upon red light irradiation. We propose that enhancement in PDT activity could be explained by their increased resistance to aggregation due to the presence of n-propyl substituent directly attached to the C6 position of the 1,4-diazepine moiety. The demonstrated results show the promising potential of tribenzo-6H-1,4-diazepinoporphyrazines as heavy atom-free photosensitizers.

Targeted locus amplification to develop robust patient-specific assays for liquid biopsies in pediatric solid tumors.

Background: Liquid biopsies combine minimally invasive sample collection with sensitive detection of residual disease. Pediatric malignancies harbor tumor-driving copy number alterations or fusion genes, rather than recurrent point mutations. These regions contain tumor-specific DNA breakpoint sequences. We investigated the feasibility to use these breakpoints to design patient-specific markers to detect tumor-derived cell-free DNA (cfDNA) in plasma from patients with pediatric solid tumors. Materials and methods: Regions of interest (ROI) were identified through standard clinical diagnostic pipelines, using SNP array for CNAs, and FISH or RT-qPCR for fusion genes. Using targeted locus amplification (TLA) on tumor organoids grown from tumor material or targeted locus capture (TLC) on FFPE material, ROI-specific primers and probes were designed, which were used to design droplet digital PCR (ddPCR) assays. cfDNA from patient plasma at diagnosis and during therapy was analyzed. Results: TLA was performed on material from 2 rhabdomyosarcoma, 1 Ewing sarcoma and 3 neuroblastoma. FFPE-TLC was performed on 8 neuroblastoma tumors. For all patients, at least one patient-specific ddPCR was successfully designed and in all diagnostic plasma samples the patient-specific markers were detected. In the rhabdomyosarcoma and Ewing sarcoma patients, all samples after start of therapy were negative. In neuroblastoma patients, presence of patient-specific markers in cfDNA tracked tumor burden, decreasing during induction therapy, disappearing at complete remission and re-appearing at relapse. Conclusion: We demonstrate the feasibility to determine tumor-specific breakpoints using TLA/TLC in different pediatric solid tumors and use these for analysis of cfDNA from plasma. Considering the high prevalence of CNAs and fusion genes in pediatric solid tumors, this approach holds great promise and deserves further study in a larger cohort with standardized plasma sampling protocols.

Characterization of the effect of chromium salts on tropocollagen molecules and molecular aggregates.

Though the capability of chromium treatment to improve the stability and mechanical properties of collagen fibrils is well-known, the influence of different chromium salts on collagen molecules (tropocollagen) is not well characterized. In this study, the effect of Cr3+ treatment on the conformation and hydrodynamic properties of collagen was studied using atomic force microscopy (AFM) and dynamic light scattering (DLS). Statistical analysis of contours of adsorbed tropocollagen molecules using the two-dimensional worm-like chain model revealed a reduction of the persistence length (i.e., the increase of flexibility) from ≈72 nm in water to ≈56-57 nm in chromium (III) salt solutions. DLS studies demonstrated an increase of the hydrodynamic radius from ≈140 nm in water to ≈190 nm in chromium (III) salt solutions, which is associated with protein aggregation. The kinetics of collagen aggregation was shown to be ionic strength dependent. Collagen molecules treated with three different chromium (III) salts demonstrated similar properties such as flexibility, aggregation kinetics, and susceptibility to enzymatic cleavage. The observed effects are explained by a model that considers the formation of chromium-associated intra- and intermolecular crosslinks. The obtained results provide novel insights into the effect of chromium salts on the conformation and properties of tropocollagen molecules.

Sliding tracheoplasty of complete tracheal cartilage rings in children.

INTRODUCTION: Complete tracheal rings are a rare malformation that occurs in 1 out of 100,000 live births. It is rare, isolated tracheal or tracheobronchial anomaly developed due to abnormal cartilage growth with formation of complete ring and often resulting in airway stenosis. Slide tracheoplasty, as it was originally described by Tsang et al. and popularized by Grillo et al., overlaps stenotic segments of trachea, shortening trachea itself, thus, doubling the circumference and diameter of the stenotic area. MATERIALS AND METHODS: We have performed slide tracheoplasty in 12 children during the period of 2019-2021 in thoracic surgery department of our center. Median age was 15 ± 21,1 months (2 months-6 years),median weight - 8,04 ± 4,75 kg (3-20,7 kg),tracheal lumen varied from 2.5 to 3.0 mm, stenosis length - from 40 to 70% of the trachea length. RESULTS: Slide tracheoplasty was performed using central veno-arterial extracorporeal membrane oxygenation in 7 cases and using cardiopulmonary bypass in 5 cases. Concomitant heart disease was revealed in 5 children (pulmonary artery sling in 3 cases, ventricular septal defects - 1, aberrant subclavian artery -1). 5 children underwent one-stage correction of VSD: plastic VSD -1; left pulmonary artery reimplantation - 3; subclavian artery reimplantation - 1. All patients were on mechanical ventilation for 4,3 ± 2,78 days at postoperative period. Patients were discharged 16,3 ± 5,14 days after surgery. Satisfactory result of treatment in the form of respiratory failure relief was achieved in 10 patients. It was possible to increase the trachea lumen from 1.5 to 2 times in all cases. There were 2 (16,6%) fatal cases due to sepsis and multi-organ failure development. CONCLUSIONS: Children with complete tracheal rings are very complicated patients with various comorbidities. Despite the advances in medicine, sometimes it is impossible to save lives of these children. The use of extracorporeal circulation (ECMO and bypass) allows us to safely perform reconstructive surgery on the trachea and save the child from respiratory failure manifestations. If needed, simultaneous correction of heart and tracheal defects is possible. Slide tracheoplasty allows to increase trachea lumen at least in 1.5-2 times. Mechanical ventilation is an unfavorable predictive factor for the outcomes of congenital tracheal stenosis management. LEVEL OF EVIDENCE: III.

Experimental Studies of the Effect of Schisandrachinensis Extract on the State of Adaptive Capabilities of Rats under Chronic and General Exposure to Cold.

Currently, there is an objective need to create fortified food products that allow not only to provide the body with energy, but also to replenish the deficiency of essential nutrients. A generalization of the information published by Rospotrebnadzor and the Institute of Nutrition of the Russian Academy of Medical Sciences indicates a deficiency in the diet of Russians of vitamins C, group B and ß-carotene and minerals, including calcium and iron, regardless of the season of the year. The identified deviations lead to a violation of the immune status, a decrease in the body's resistance to infections, and other unfavorable environmental factors, leading to an increase in the level of morbidity and a decrease in working capacity. The main unfavorable climatic factor that the population of the Far Eastern region has to face is low freezing temperatures. Adaptation to cold exposure is a complex process that requires a long period and may be accompanied by functional disorders and morphological changes in body tissues. In connection with the above, the problem of increasing the adaptive capabilities of a person to unfavorable environmental factors by means of correcting daily nutrition, providing the body with essential macro- and micronutrients, which is important in the prevention of possible diseases, is of particular importance. This study is aimed at assessing the effect of Schisandrachinensis extract on the adaptive capacity of rats in conditions of chronic and general cold. It was found that the extracts obtained from the fruits of Schisandra chinensis are characterized by a high content of biologically active substances. In experiments with determining the duration of running on the treadmill, a distinct act-protective effect was observed with the introduction of Schisandra chinensis extracts at a dose of 150 mg/day, against the background of reduced resistance to physical activity due to cold exposure. It was found that exposure to cold significantly reduced the swimming resistance of rats on all days of the study. The introduction of Schisandra chinensis extract into the diet led to an increase in resistance to fatigue and an increase in the duration of swimming on all days of the experiment. Conclusions: in this experimental model, a gradually increasing effect of increasing the physical performance of rats was demonstrated with prolonged (28 days) intake of the developed drinks, which coincides with the literature data on a number of other adaptogens and indicates the presence of cumulative properties of biologically active substances of Schisandra extract.

Minimal residual disease (MRD) detection in acute lymphoblastic leukaemia based on fusion genes and genomic deletions: towards MRD for all.

Minimal residual disease (MRD) diagnostics are implemented in most clinical protocols for patients with acute lymphoblastic leukaemia (ALL) and are mostly performed using rearranged immunoglobulin (IG) and/or T-cell receptor (TR) gene rearrangements as molecular polymerase chain reaction targets. Unfortunately, in 5-10% of patients no or no sensitive IG/TR targets are available, and patients therefore cannot be stratified appropriately. In the present study, we used fusion genes and genomic deletions as alternative MRD targets in these patients, which retrospectively revealed appropriate MDR stratification in 79% of patients with no (sensitive) IG/TR target, and a different risk group stratification in more than half of the cases.

Sea Buckthorn and Rosehip Oils with Chokeberry Extract to Prevent Hypercholesterolemia in Mice Caused by a High-Fat Diet In Vivo.

Dietary supplementation based on sea buckthorn and rosehip oils with added chokeberry extract was studied. We added the dietary supplement to the feed mixtures for laboratory animals. The possible toxicological effects and hypocholesterolemic, hepatoprotective activity of the dietary supplement in vivo were studied. After the observation period (6 weeks), no significant changes were found in the mass of organs and blood serum of laboratory animals (p > 0.05). However, there was a decrease in hypercholesterolemic indicators. Regular consumption of sea buckthorn and rosehip oils with added chokeberry extract (dietary supplement "ESB-1") by laboratory animals inhibited the activity of liver enzymes and increased the antioxidant activity of blood serum (after the subcutaneous injection of sunflower oil/oil solution of carbon tetrachloride) but was not sufficient to bring them to physiological standards. The hypocholesterolemic and antioxidant properties of our dietary supplement already allow us to consider it a component of functional food products or a dietary supplement base. However, the full range of its biologically active properties, including the hepatoprotective function and regulation of metabolic disorders, has not been studied yet, which sets the direction of further research in vivo models and clinical practice to confirm its effectiveness in humans.

Blastic Plasmacytoid Dendritic Cell Neoplasm: Case Report and Literature Overview.

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a malignancy with high frequency of skin involvement. A 39-year-old Caucasian female was suffering from weakness, myalgia, and skin eruption, which appeared during treatment of chlamydiosis with antibiotics in July 2016. Based on clinical presentation, laboratory investigations, and histological examination of skin and bone marrow biopsy, a diagnosis of BPDCN with the involvement of skin, bone marrow, and central nervous system was made. The patient was put on acute lymphoblastic leukemia-like chemotherapy and achieved complete remission in November 2016, the eruption regressed. In January 2017, allogeneic bone marrow transplantation from matched sibling was performed. Since May 2017, the cutaneous relapse with loss of CD56 expression has developed. This clinical case demonstrates the importance of laboratory tests. Histological examination helps to clarify a diagnosis of cutaneous lymphoma; however, a specific type of lymphoma needs immunohistochemical analysis. In our case, BPDCN at the initial stage presented like a systemic vasculitis.

Ultrasonography patterns of atopic dermatitis in children.

BACKGROUND: Atopic dermatitis (AD) is one of the most common allergic diseases in children. The aim of the study was to evaluate the ultrasound picture of lesional and non-lesional skin in children with AD. MATERIALS AND METHODS: The study included a group of children with AD and a control group. Inclusion criteria were as follows: age 0-8 years and clinical diagnosis of AD. An ultrasound scanner with a 75 MHz transducer probe was used to produce B-mode skin images in lesions and non-lesional skin. The thickness and the echogenicity of epidermis, dermis, and subepidermal low-echogenic band (SLEB) were measured, and the ratio coefficient per body site was calculated. RESULTS: Ultrasonography of non-lesional skin in children with AD showed uneven epidermis contour, a tendency to increased epidermis and decreased dermis thickness, and the SLEB was observed in 77% of cases. In lesions, there was an increased thickness and a decreased echogenicity of epidermis and dermis, and epidermis had irregular contours in most cases. The SLEB was in all lesions, showing greater thickness and lower echogenicity compared with non-lesional skin. CONCLUSION: HF-USG of the skin allows visualizing the epidermal barrier disruption and inflammation in dermis in children with AD on the entire surface of the skin.

A Female Patient with FMR1 Premutation and Mosaic X Chromosome Aneuploidy and Two Sons with Intellectual Disability.

In this report, we describe a molecular cytogenetic study of a family burdened with intellectual disability (ID) and suicide. Our study revealed that the mother has a heterozygous premutation in the FMR1 gene and supernumerary X chromosomes as well as X-derived marker chromosomes. Both of her sons have ID and a normal chromosome number. One of the sons has fragile X syndrome, and the other has ID of an unclear nature.

Identification of a regulatory domain controlling the Nppa-Nppb gene cluster during heart development and stress.

The paralogous genes Nppa and Nppb are organized in an evolutionarily conserved cluster and provide a valuable model for studying co-regulation and regulatory landscape organization during heart development and disease. Here, we analyzed the chromatin conformation, epigenetic status and enhancer potential of sequences of the Nppa-Nppb cluster in vivo Our data indicate that the regulatory landscape of the cluster is present within a 60-kb domain centered around Nppb Both promoters and several potential regulatory elements interact with each other in a similar manner in different tissues and developmental stages. The distribution of H3K27ac and the association of Pol2 across the locus changed during cardiac hypertrophy, revealing their potential involvement in stress-mediated gene regulation. Functional analysis of double-reporter transgenic mice revealed that Nppa and Nppb share developmental, but not stress-response, enhancers, responsible for their co-regulation. Moreover, the Nppb promoter was required, but not sufficient, for hypertrophy-induced Nppa expression. In summary, the developmental regulation and stress response of the Nppa-Nppb cluster involve the concerted action of multiple enhancers and epigenetic changes distributed across a structurally rigid regulatory domain.

A transgenic mouse model for the simultaneous monitoring of ANF and BNP gene activity during heart development and disease.

AIM: The expression of Nppa (ANF) and Nppb (BNP) marks the chamber myocardium in the embryo, and both genes serve as early and accurate markers for hypertrophy and heart failure. Non-invasive visualization of Nppa-Nppb expression in living mice would enable to evaluate the disease state during the course of time in heart disease models. We sought to develop a method to assess the pattern and level of Nppa and Nppb expression within living mice. METHODS AND RESULTS: A modified bacterial artificial chromosome containing a genomic segment spanning the Nppa-Nppb locus was randomly integrated into the mouse genome. Firefly Luciferase was inserted into Nppa and the red fluorescent protein gene Katushka into Nppb. Both reporters precisely recapitulated the spatio-temporal patterns of Nppa and Nppb, respectively. In a hypertrophy model (transverse aortic constriction) and myocardial infarction model (left anterior descending coronary artery occlusion), the non-invasively measured bioluminescent signal from Luciferase correlated with Nppa expression, and the intensity of red fluorescence with levels of the expression of Katushka and Nppb. After myocardial infarction, the border zone of the infarct area was readily identified by an increased intensity of Katushka fluorescence. CONCLUSIONS: A genomic region sufficient to regulate the developmental pattern and stress response of Nppa and Nppb has been defined. The double reporter mice can be used for the functional imaging and investigation of cardiac hypertrophy and myocardial infarction in vivo.

Regulation of expression of atrial and brain natriuretic peptide, biomarkers for heart development and disease.

The mammalian heart expresses two closely related natriuretic peptide (NP) hormones, atrial natriuretic factor (ANF) and brain natriuretic peptide (BNP). The excretion of the NPs and the expression of their genes strongly respond to a variety of cardiovascular disorders. NPs act to increase natriuresis and decrease vascular resistance, thereby decreasing blood volume, systemic blood pressure and afterload. Plasma levels of BNP are used as diagnostic and prognostic markers for hypertrophy and heart failure (HF), and both ANF and BNP are widely used in biomedical research to assess the hypertrophic response in cell culture or the development of HF related diseases in animal models. Moreover, ANF and BNP are used as specific markers for the differentiating working myocardium in the developing heart, and the ANF promoter serves as platform to investigate gene regulatory networks during heart development and disease. However, despite decades of research, the mechanisms regulating the NP genes during development and disease are not well understood. Here we review current knowledge on the regulation of expression of the genes for ANF and BNP and their role as biomarkers, and give future directions to identify the in vivo regulatory mechanisms. This article is part of a Special Issue entitled: Heart failure pathogenesis and emerging diagnostic and therapeutic interventions.