Physiological Conditions Research Articles

Dithiothreitol prevents the spontaneous release of cortical granules in in vitro aged mouse oocytes by protecting regulatory proteins of cortical granules exocytosis and thickening the cortical actin cytoskeleton

In assisted fertility protocols, in vitro culture conditions mimic physiological conditions to preserve gametes in the best conditions. After collection, oocytes are maintained in a culture medium inside the incubator until in vitro fertilization (IVF) is performed. This time outside natural and physiological conditions exposes oocytes to an oxidative stress that renders in vitro aging. It has been described that in vitro aging produces a spontaneous cortical granule (CG) release decreasing the fertilization rate of oocytes. Nevertheless, this undesirable phenomenon has not been investigated, let alone prevented. In this work, we characterized the spontaneous CG secretion in in vitro aged oocytes. Using immunofluorescence indirect, quantification, and functional assays, we showed that the expression of regulatory proteins of CG exocytosis was affected. Our results demonstrated that in vitro oocyte aging by 4 and 8 h altered the expression and localization of alpha-SNAP and reduced the expression of NSF and Complexin. These alterations were prevented by supplementing culture medium with dithiothreitol (DTT), which in addition to having a protective effect on those proteins, also had an unexpected effect on the actin cytoskeleton. Indeed, DTT addition thickened the cortical layer of fibrillar actin. Both DTT effects, together, prevented the spontaneous secretion of CG and recovered the IVF rate in in vitro aged oocytes. We propose the use of DTT in culture media to avoid the spontaneous CG secretion and to improve the success rate of IVF protocols in in vitro aged oocytes.

Gelatin-based carbon quantum dot-molecularly imprinted polymer: Safe photoluminescent core-shell nano-carrier for the pH-responsive anticancer drug delivery

This study aims to synthesize a core-shell gelatin-based carbon quantum dot-molecularly imprinted polymer (MIP@g-CQD) via the precipitation free-radical polymerization process using methotrexate (MTX) as a model anticancer template. To investigate the efficiency of the prepared photoluminescent MIP@g-CQD as a pH-responsive nano-carrier, MTX was loaded into MIP@g-CQD by soaking in a drug solution and the release behavior of the loaded drug was evaluated in the necessary pH values (7.4, 5). The successful synthesis of materials was characterized using PL, TEM, FE-SEM, DLS, and FT-IR analyses. Interestingly, the created cavities in the core-shell nano-carriers can interact with the MTX molecules effectively, leading to an increase in the loading capacity. According to the obtained results from Langmuir adsorption isotherms, the imprinting factor was calculated (IF = 4.91). Also, the binding kinetics of MTX revealed the creation of particular recognition sites in the core-shell polymeric network. The MTX-loaded MIP@g-CQD displayed a low rate and limited release at the simulated physiological environment (pH 7.4, 37 °C), but it is increased at tumor tissue (pH 5, 41 °C) conditions, which can lead to long-term and sustained release of MTX in the desired target. This property of MIP@g-CQD could avoid the release of MTX in normal physiological conditions, decreasing the possible side effects of MTX drug. Owing to the existence of amide functional groups in the nano-carrier structure and its negatively charged nature, the MTT assay displayed desirable cytotoxicity against the breast cancer cell line (MCF-7) for the MTX-loaded nano-carrier. According to the obtained results, the prepared safe photoluminescent MIP@g-CQD with appropriate pH-responsivity has a high ability to be applied as an anticancer and bio-detection agent.

Preparation of bacterial cellulose/acrylic acid-based pH-responsive smart dressings by graft copolymerization method

Conventional wound dressings used in trauma treatment have a single function and insufficient adaptability to the wound environment, making it difficult to meet the complex demands of the healing process. Stimuli-responsive hydrogels can respond specifically to the particular environment of the wound area and realize on-demand responsive release by loading active substances, which can effectively promote wound healing. In this paper, BC/PAA-pH responsive hydrogels (BPPRHs) were prepared by graft copolymerization of acrylic acid (AA) to the end of the molecular chain of bacterial cellulose (BC) network structure. Antibacterial pH-responsive ‘smart’ dressings were prepared by loading curcumin (Cur) onto the hydrogels. Surface morphology, chemical groups, crystallinity, rheological, and mechanical properties of BPPRHs were analyzed by different characterization methods. The drug release behavior under different physiological conditions and bacteriostatic properties of BPPRH-Cur dressings were also investigated. The results of structural characterization and performance studies show that the hydrogel has a three-dimensional mesh structure and can respond to wound pH in a ‘smart’ drug release capacity. The drug release behavior of the BPPRH-Cur dressings under different environmental conditions conformed to the logistic and Weibull kinetic models. BPPRH-Cur displayed good antimicrobial activity against common pathogens of wound infections such as E. coli, S. aureus, and P. aeruginosa by destroying the cell membrane and lysing the bacterial cells. This study lays the foundation for the development of new pharmaceutical dressings with positive health, economic and social benefits.

Oncoviral Infections and Small Extracellular Vesicles.

Small extracellular vesicles (sEV) are small membrane-bound nanovesicles with a size range below 200 nm that are released by all types of cells. sEV carry a diverse cargo of proteins, lipids, glycans, and nucleic acids that mimic the content of producer cells. sEV mediate intercellular communication and play a key role in a broad variety of physiological and pathological conditions. Recently, numerous reports have emerged examining the role of sEV in viral infections. A significant number of similarities in the sEV biogenesis pathways and the replication cycles of viruses suggest that sEV might influence the course of viral infections in diverse ways. Besides directly modulating virus propagation by transporting the viral cargo (complete virions, proteins, RNA, and DNA), sEV can also modify the host antiviral response and increase the susceptibility of cells to infection. The network of mutual interactions is particularly complex in the case of oncogenic viruses, deserving special consideration because of its significance in cancer progression. This review summarizes the current knowledge of interactions between sEV and oncogenic viruses, focusing on sEV abilities to modulate the carcinogenic properties of oncoviruses.

Optimization of polymer coating thickness and strut width in drug Eluting stents using Magnetic field

The transport of drug/magnetic particle (MP) conjugates in the presence of a Magnetic Field (MF) in Drug Eluting Stents (DESs) is modeled numerically using the Finite Volume Method (FVM). The effects of physiological conditions corresponding to different degrees of calcification, drug particles sizes and hematocrit level, were analyzed by investigating the roles of the tissue permeability, its anisotropy and the plasma viscosity. It was found that both in the absence and presence of the MF, as the tissue permeability decreases or the plasma viscosity increases, the free-phase drug and Extracellular Matrix (ECM)-bound phase contents increase. Stronger tissue anisotropy leads to a decrease of the free-phase drug content and an increase of the ECM-bound phase content. Within the explored ranges, the Specific Receptor (SR)-bound phase of the drug was found to be insensitive to the tissue permeability and plasma viscosity, and to be larger in anisotropic tissues. The activation of the MF leads systematically to larger free-phase drug contents, with the increases most prominent at smaller tissue permeability, anisotropy and plasma viscosity. On the other hand, the effects on the ECM-bound phase content are found to be stronger at larger permeability, smaller plasma viscosity and lower tissue anisotropy. For an isotropic tissue, the MF induces a decrease of the ECM-bound phase content at early times, followed by an increase at later times. For the considered ranges of permeability and viscosity, the MF does not seem to have any noticeable effects on the SR-bound phase. However, this phase of the drug tends to increase with the activation of the MF in isotropic tissues and is unchanged in anisotropic ones. These reported effects of the MF hold promise for alleviating two factors contributing to In-Stent Restenosis, namely the polymer coating width and thickness. The study reveals that a narrower or thinner polymer layer, in combination with the MF, can mimic the drug release dynamics of a wider or thicker polymer layer in the absence of the MF. The corresponding width and thickness of the magnetized stents, that we referred to as the equivalent polymer width (EPW) and equivalent polymer thickness (EPT) were determined and their dependence on the tissue permeability, isotropy and the plasma viscosity, was investigated. The study shows that it is possible to achieve the same drug delivery with polymer coating of half the width or half the thickness of the non-magnetized stent when an electric intensity of 3A is used.

Biochar Improves Yield by Reducing Saline-Alkaline Stress, Enhancing Filling Rate of Rice in Soda Saline-Alkaline Paddy Fields.

Soda saline-alkaline stress significantly impedes the rice grain filling process and ultimately impacts rice yield. Biochar has been shown to mitigate the negative impacts of saline-alkaline stress on plants. However, the exact mechanism by which biochar influences the rice grain-filling rate in soda saline-alkaline soil is still not fully understood. A two-year field experiment was conducted with two nitrogen fertilizer levels (0 and 225 kg ha-1) and five biochar application rates [0% (B0), 0.5% (B1), 1.5% (B2), 3.0% (B3), and 4.5% (B4) biochar, w/w]. The results demonstrated that biochar had a significant impact on reducing the Na+ concentration and Na+/K+ ratio in rice grown in soda saline-alkaline lands, while also improving its stress physiological conditions. B1, B2, B3, and B4 showed a notable increase in the average grain-filling rate by 5.76%, 6.59%, 9.80%, and 10.79%, respectively, compared to B0; the time to reach the maximum grain-filling rate and the maximum grain weight saw increases ranging from 6.02% to 12.47% and from 7.85% to 14.68%, respectively. Meanwhile, biochar, particularly when used in conjunction with nitrogen fertilizer, notably enhanced the activities of sucrose synthase (SuSase), ADPG pyrophosphorylase (AGPase), starch synthase (StSase), and starch branching enzyme (SBE) of rice grains in soda saline-alkaline lands. Furthermore, rice yield increased by 11.95-42.74% in the B1, B2, B3, and B4 treatments compared to the B0 treatment. These findings showed that biochar improves yield by regulating ionic balance, physiological indicators, starch synthesis key enzyme activities, and the grain-filling rate in soda saline-alkaline paddy fields.

Experimental and computational perspectives on the interaction of nerve agent VX metabolite ethyl methylphosphonic acid with human serum albumin

This study employed electrochemical methods to investigate the interaction between ethyl methylphosphonic acid (EMPA), the primary hydrolysis product of the nerve agent VX, and human serum albumin (HSA). Utilizing cyclic voltammetry (CV) and differential pulse voltammetry (DPV) on the surface of a glassy carbon electrode (GCE). The interaction was analyzed based on changes in the electrochemical behavior of the K3Fe(CN)6/K4Fe(CN)6 redox species under physiological pH conditions. Electrochemical parameters of the redox probe in the presence of HSA, EMPA, and the EMPA-HSA complex were measured. The alteration of the redox couple responses indicates the formation of an electro-inactive complex between the protein and EMPA. The binding parameters of the EMPA-HSA interaction, including the binding constant and complex stoichiometry, were determined, revealing the formation of EMPA-HSA complex with a binding constant of 1.03 × 104M−1 and a binding number of 0.624, suggesting a 1:1 stoichiometry. Molecular modeling was also conducted to further investigate the binding mode of EMPA with HSA, where the binding energy of EMPA to HSA was calculated as −4.78 kcal/mol with hydrogen bonding and hydrophobic contacts being the main interactions behind the complex formation. This research offers valuable insights into the albumin-EMPA interaction, with implications for toxicology, pharmacokinetics, and environmental impact.

Dual drug-loaded tumor-targeted polymeric nanoparticles for enhancing therapeutic response in pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease where standard-of-care chemotherapeutic drugs have limited efficacy due to the development of drug resistance and poor drug delivery caused by a highly desmoplastic tumor microenvironment. Combining multiple drugs in a tumor-targeting carrier would be a favorable approach to overcome these limitations. Hence, a tumor-targeted peptide (TTP) conjugated amphiphilic tri-block copolymer was developed to make targeted polymer nanoparticles (TTP-PNPs) serving as a vehicle for carrying gemcitabine (Gem), paclitaxel (PTX), and their combination (Gem + PTX). The TTP-PNPs in the form of empty polymer (P), single drug-loaded [P(Gem) and P(PTX)], and dual drug-loaded [P(Gem + PTX)] polymer nanoformulations exhibited stable and homogenous spherical shapes with 110–160 nm size. These nanoformulations demonstrated excellent stability under in vitro physiological conditions and led to an efficient release of the drugs in the presence of reduced glutathione (GSH). The efficacy of these nanoparticles was thoroughly evaluated in vitro and in vivo, demonstrating a notable capacity to selectively target and restrict PDAC cells (PANC-1 and KPC) growth. The cellular uptake and biodistribution study showed a significantly higher tumor-targeting ability of TTP-PNPs than PNPs without TTP. Notably, P(Gem + PTX) exhibited the lowest IC50 compared to all other controls and showed heightened synergistic effects in both cell lines. Furthermore, P(Gem + PTX) showed a significantly better tumor reduction and median overall survival in mouse models than single drug-loaded TTP-PNPs or a combination of free drugs (Gem + PTX). In summary, our TTP-PNP system shows great promise as a novel platform for delivering Gem + PTX specifically to pancreatic cancer (PC), maximizing the therapeutic benefits with lower concentrations of the drugs and potentially reducing toxic side effects.

TNF Induces Laminin-332-Encoding Genes in Endothelial Cells and Laminin-332 Promotes an Atherogenic Endothelial Phenotype.

Laminins are essential components of the basement membranes, expressed in a tissue- and cell-specific manner under physiological conditions. During inflammatory circumstances, such as atherosclerosis, alterations in laminin composition within vessels have been observed. Our study aimed to assess the influence of tumor necrosis factor-alpha (TNF), a proinflammatory cytokine abundantly found in atherosclerotic lesions, on endothelial laminin gene expression and the effects of laminin-332 (LN332) on endothelial cells' behavior. We also evaluated the expression of LN332-encoding genes in human carotid atherosclerotic plaques. Our findings demonstrate that TNF induces upregulation of LAMB3 and LAMC2, which, along with LAMA3, encode the LN332 isoform. Endothelial cells cultured on recombinant LN332 exhibit decreased claudin-5 expression and display a loosely connected phenotype, with an elevated expression of chemokines and leukocyte adhesion molecules, enhancing their attractiveness and adhesion to leukocytes in vitro. Furthermore, LAMB3 and LAMC2 are upregulated in human carotid plaques and show a positive correlation with TNF expression. In summary, TNF stimulates the expression of LN332-encoding genes in human endothelial cells and LN332 promotes an endothelial phenotype characterized by compromised junctional integrity and increased leukocyte interaction. These findings highlight the importance of basement membrane proteins for endothelial integrity and the potential role of LN332 in atherosclerosis.

Engineered Carbon Nanocatalysts for Homocysteine Catabolism

Cystathionine-β-synthase (CBS) deficiency or homocystinuria (HCU) is a rare inherited disorder with main clinical features of osteoporosis, dislocation of the optic lenses, learning difficulties and thromboembolism. CBS is a key enzyme in methionine catabolism pathway. CBS deficiency impairs the conversion of homocysteine (Hcy) to cystathionine, which leads to extracellular buildup of Hcy in toxic concentrations. Patients with CBS deficiency have elevated plasma Hcy and methionine.Currently, there is no FDA approved drug for CBS deficiency. Clinical treatment options are low-protein diet and betaine for severely affected patients, and vitamin supplementation for mildly affected adults. Artificial protein-based enzymes for Hcy treatment are in clinical trials. However, protein-based artificial enzymes can be immunogenic, prone to intracellular modifications, and the treatment may not be viable after few months.We propose a novel nanotechnology-based approach for the treatment of CBS deficiency. Carbon nanomaterials with multiple unsaturated carbons provide an excellent scaffold for engineering enzyme mimicking catalysts. We have designed carbon nanomaterial based catalytic artificial enzymes (CAEs) by surface engineering to mimic metabolic enzymes that catabolize Hcy under physiological conditions. Using a combination of paper chromatography and H1-NMR, our lab has shown that CAE can breakdown homocysteine and produce hydrogen sulfide similar to CGL enzyme. The degree of surface functionalization as well as surface curvature has profound effect on specificity and catalysis. We have shown that CAE can enter cells and lower homocysteine levels in cellular models. In preliminary studies, CAE was able to lower serum homocysteine levels in a murine model. Further studies will involve safety assessments of CAE as a treatment for HCU.

(Invited) An in Vitro Model of Mechanical Strain Enhances Cellular Uptake of Single-Walled Carbon Nanotubes

Single-walled carbon nanotubes (SWCNTs) are ideal optical biosensors due to their intrinsic near-infrared (NIR) emission, which is highly sensitive, easily tunable, and indefinitely photostable. Due to such desirable properties coupled with appropriate functionalization strategies, SWCNTs have been used in numerous in vitro, live-cell, and in vivo applications. Depending on the type of functionalization as well as cell type, SWCNTs can be internalized by live cells via demonstrated energy-dependent endocytosis processes where they are retained within the endosomal pathway or are expelled by the cells. While this uptake and intracellular processing of SWCNTs has been recently examined, these studies are usually performed with standard cell culture techniques in petri dishes or 3D cultures. Unlike these cultures, cells in living organisms are continuously exposed to a wide variety of stimuli, e.g., periodic mechanical strain, during both normal and disease-related conditions. Thus, it is important to investigate how cells under physiological stretching compare to those grown in standard cultures. Herein, using a novel cell stretching platform to mimic physiological conditions, we investigate the uptake of SWCNTs into different human cancer cells.Our results indicate that cells exposed to uniform radial stretching display significantly higher rates of nanoparticle uptake, which can be correlated to their increased level of cellular activity. This approach recognizes the importance of studying cellular responses in environments that more closely resemble the complex conditions found in living organisms. It suggests a potential connection between mechanical stimuli and cellular processes related to SWCNT uptake. This finding has implications for understanding how nanotube uptake differs in different cellular stress environments and how cells respond to their dynamic and ever-changing microenvironment.Investigating how the mechanical strain influences the endocytosis processes and subsequent intracellular fate of SWCNTs can provide valuable insights into the interplay between nanomaterials and living cells under more realistic conditions. Further aiding us in the development of more accurate intracellular biosensors and effective drug delivery platforms using SWCNTs.

Electrochemical and Spectroscopic Investigations of Bismuth(III) Pharmaceuticals with L-Glutathione

Due to the importance of L-cysteine in the human system, the interaction of metals with this amino acid are worthy of investigation by various methods. In this laboratory1, we have investigated the interaction of bismuth(III) compounds with L-cysteine by both cyclic voltammetry and UV-VIS spectroscopy in an attempt to understand the role of bismuth in treating gastrointestinal maladies.2 The addition of bismuth(III) compounds to solutions of L-cysteine causes a negative potential shift for bismuth(III) reduction as an indication of complex formation between bismuth(III) and L-cysteine. This interaction is also evident by the appearance of a UV-VIS spectroscopic band at 340 nm (Bi-sulfur bond). Even at low pH values (pH 1.0), electrochemical studies show evidence of some kind of interaction even when there is no 340 nm band. These investigations have been conducted at pH 1.00 and 3.00 to simulate conditions in the human stomach and at pH 7.40 MOPS [(3-N-morpholino)propanesulfonic acid] to simulate general physiological conditions, in particular the intestinal tract. Recent work has involved extension of this work to L-glutathione, a tripeptide incorporating L-cysteine, under similar conditions. As might be expected, these interactions are considerably more complicated than those involving L-cysteine. As an example, there is a considerable positive potential shift for the reduction of bismuth(III) citrate in the presence of L-glutathione on the first and second voltammetric sweeps at gold and glassy carbon electrodes. This behavior evidently reflects the rather long (several seconds) time required for the re-formation of the bismuth complex with L-glutathione after its reductive ligand (L-glutathione) loss during bismuth(III) reduction on the first sweep. In addition, there is evidence of a film buildup on the electrode surface as observed by peak current increases during multiple potential sweeps. These observations are planned for inclusion in this symposium presentation.References T. Cheek and D. Peña, J. Electrochem. Soc., 167, 155522 (2020)Li, R. Wang, and H. Sun, Acc. Chem. Res., 52, 216 (2019).

Ion-channel degeneracy and heterogeneities in the emergence of signature physiological characteristics of dentate gyrus granule cells.

Complex systems are neither fully determined nor completely random. Biological complex systems, including single neurons, manifest intermediate regimes of randomness that recruit integration of specific combinations of functionally segregated subsystems. Such emergence of biological function provides the substrate for the expression of degeneracy, the ability of disparate combinations of subsystems to yield similar function. Here, we present evidence for the expression of degeneracy in morphologically realistic models of dentate gyrus granule cells (GC) through functional integration of disparate ion-channel combinations. We performed a 45-parameter randomized search spanning 16 active and passive ion channels, each biophysically constrained by their gating kinetics and localization profiles, to search for valid GC models. Valid models were those that satisfied 17 sub- and supra-threshold cellular-scale electrophysiological measurements from rat GCs. A vast majority (>99%) of the 15,000 random models were not electrophysiologically valid, demonstrating that arbitrarily random ion-channel combinations wouldn't yield GC functions. The 141 valid models (0.94% of 15,000) manifested heterogeneities in and cross-dependencies across local and propagating electrophysiological measurements, which matched with their respective biological counterparts. Importantly, these valid models were widespread throughout the parametric space and manifested weak cross-dependencies across different parameters. These observations together showed that GC physiology could neither be obtained by entirely random ion-channel combinations nor is there an entirely determined single parametric combination that satisfied all constraints. The complexity, the heterogeneities in measurement and parametric spaces, and degeneracy associated with GC physiology should be rigorously accounted for, while assessing GCs and their robustness under physiological and pathological conditions.

The Pivotal Role of the Key Angiogenic Factors in the Development of Endometrioid Pathologies of the Uterus and Ovary.

A characteristic feature of uterine pathologies is a specific change in cell metabolism, which predominantly manifests as a shift in the need for nutrients, thereby directing cells to engage in different angiogenic marker activities. Angiogenesis is one of the main signals supporting the survival and development of cells and tissues not only under physiological conditions. Therefore, it is necessary that we understand pathological hyperactivation in all uterine diseases, from endometriosis through ovarian endometrioid adenocarcinoma to malignant transformed cells of the uterine epithelium and body. This work presents the gene expression results of selected angiogenesis targets (VEGF-A, TGF-β1, ANG1/2, and HIF-1α), cell migration, and cell-cell interaction determined in vitro. Our results suggest that angiogenesis varies in the tested pathological conditions (ectopic endometriosis-12Z; ovarian endometrioid adenocarcinoma-A2780; tumors-SK-UT-1 and RL-95-2) compared to physiological angiogenesis (HME1). The differential expression of angiogenic factors may contribute (or is a contributing factor) to the observed differences to acknowledge an inherent variability in angiogenesis among cell lines. Determining the genomic phenomena responsible for processes associated with inadequate angiogenesis in the pelvic region could help us to develop individual treatment strategies and explain resistance to treatment.

Uncovering the mechanism of troglitazone-mediated idiosyncratic drug-induced liver injury with individual-centric models.

Idiosyncratic drug-induced liver injury is a rare and unpredictable event. Deciphering its initiating-mechanism is a hard task as its occurrence is individual dependent. Thus, studies that utilize models that are not individual-centric might drive to a general mechanistic conclusion that is not necessarily true. Here, we use the individual-centric spheroid model to analyze the initiating-mechanism of troglitazone-mediated iDILI risk. Individual-centric spheroid models were generated using a proprietary cell educating technology. These educated spheroids contain hepatocytes, hepatic stellate cells, activated monocyte-derived macrophages, and dendritic cells under physiological conditions. We show that phases 1 and 2 drug-metabolizing enzymes were induced in an individual-dependent manner. However, we did not observe any association of DEMs induction and troglitazone (TGZ)-mediated iDILI risk. We analyzed TGZ-mediated iDILI and found that a 44-year-old male showed iDILI risk that is associated with TGZ-mediated suppression of IL-12 expression by autologous macrophages and dendritic cells. We performed a rescue experiment and showed that treatment of spheroids from this 44-year-old male with TGZ and recombinant IL-12 suppressed iDILI risk. We confirmed the mechanism in another 31-year-old female with iDILI risk. We demonstrate here that individual-centric spheroid are versatile models that allow to predict iDILI risk and to analyze a direct effect of the drug on activated macrophages and dendritic cells to uncover the initiating-mechanism of iDILI occurrence. This model opens perspectives for a personalized strategy to mitigate iDILI risk.

Alternative drug delivery systems for pregnancy related disorders: A focused review

Objectives: The objective of this review was to summarize currently developed advanced drug delivery systems that can be used for pregnancy related disorders along with their advantages and clinical relevance. Methods: This process used a defined search strategy of indexed publications from databases like Google scholar, PubMed, Embase, Medline, Scopus and UTMB library from 2000 to 2020 and included methods to assess the quality of the evidence retrieved. Results: Pregnancy is the unique physiological condition where some of the drugs will be lifesaving to mother, but they can be toxic to fetal tissue. It is mandatory to research on the novel drug delivery systems. Although there was a substantial quantity of research has been carried out on the maternal -placental-fetal drug uptake, transfer, and toxicity, therapeutic evaluations of advanced drug delivery systems in pregnancy remain in their infancy. Conclusions: In this review, we highlighted the physiological considerations of the placenta and summarized recent research work in advanced drug delivery systems used in pregnancy as well as their potential applications in clinical reality.

Chronic administration of prebiotics and probiotics ameliorates pathophysiological hallmarks of Alzheimer’s disease in a APP/PS1 transgenic mouse model

Introduction: The gut microbiota (MB), although one of the main producers of Aβ in the body, in physiological conditions contributes to the maintainance of a healthy brain. Dysbiosis, the dysbalance between Gram-negative and Gram-positive bacteria in the MB increases Aβ production, contributing to the accumulation of Aβ plaques in the brain, the main histopathological hallmark of Alzheimer’s disease (AD). Administration of prebiotics and probiotics, maintaining or recovering gut-MB composition, could represent a nutraceutical strategy to prevent or reduce AD sympthomathology. Aim of this research was to evaluate whether treatment with pre- and probiotics could modify the histopathological signs of neurodegeneration in hippocampal CA1 and CA3 areas of a transgenic mouse model of AD (APP/PS1 mice). The hippocampus is one of the brain regions involved in AD.Methods: Tg mice and Wt littermates (Wt-T and Tg-T) were fed daily for 6 months from 2 months of age with a diet supplemented with prebiotics (a multi-extract of fibers and plant complexes, containing inulin/fruit-oligosaccharides) and probiotics (a 50%–50% mixture of Lactobacillus rhamnosus and Lactobacillus paracasei). Controls were Wt and Tg mice fed with a standard diet. Brain sections were immunostained for Aβ plaques, neurons, astrocytes, microglia, and inflammatory proteins that were evaluated qualitatively and quantitatively by immunofluorescence, confocal microscopy and digital imaging with ImageJ software.Results: Quantitative analyses demonstrated that: 1) The treatment with pre- and probiotics significantly decreased Aβ plaques in CA3, while in CA1 the reduction was not significant; 2) Neuronal damage in CA1 Stratum Pyramidalis was significantly prevented in Tg-T mice; no damage was found in CA3; 3) In both CA1 and CA3 the treatment significantly increased astrocytes density, and GFAP and IBA1 expression, especially around plaques; 4) Microglia reacted differently in CA1 and CA3: in CA3 of Tg-T mice there was a significant increase of CD68+ phagocytic microglia (ball-and-chain phenomic) and of CX3CR1 compared with CA1.Discussion: The higher microglia reactivity could be responsible for their more efficient scavenging activity towards Aβ plaques in CA3 in comparison to CA1. Treatment with pre- and probiotics, modifying many of the physiopathological hallmarks of AD, could be considered an effective nutraceutical strategy against AD symptomatology.

Redox Regulation of K+ channel: Role of Thioredoxin.

Potassium channels regulate the influx and efflux of K+ ions in various cell types that generate and propagate action potential associated with excitation, contraction and relaxation of various cell types. Although redox active cysteines are critically important for channel activity, redox regulation of K+ channels by thioredoxin has not been systematically reviewed. Redox regulation of K+ channel is now increasingly recognized as drug targets in pathological condition of several cardiovascular disease processes. The role of thioredoxin in regulation of these channels and its implication in pathological conditions have not been adequately reviewed. This review specifically focuses on the redox-regulatory role of thioredoxin on K+ channel structure and function in physiological and pathophysiological conditions. Ion channels including K+ channel have been implicated in the functioning of cardiomyocyte excitation-contraction coupling, vascular hyperpolarization, cellular proliferation, and neuronal stimulation in physiological and pathophysiological conditions. Although, oxidation-reduction of ion channels is critically important in their function, role of thioredoxin, redox regulatory protein in regulation of these channels and its implication in pathological conditions need to be studied to gain further insight into channel function. Future studies need to map all redox regulatory pathways in channel structure and function using novel mouse models, redox proteomic and signal transduction studies, which modulate various currents and altered excitability of relevant cells implicated in a pathological condition. We are yet at infancy of studies related to redox control of various K+ channels and structured and focused studies with novel animal models.

Structural and biophysical insights into the interactions of anisotropic gold nanoparticles with human telomeric G-quadruplex DNA: Spectroscopic and calorimetric approach

We are reporting curcumin-induced synthesis of anisotropic citrate capped Gold nanoparticles (ctGNPs). The techniques such as UV–visible spectroscopy, Raman spectroscopy, FT-IR, X-ray diffraction (XRD), and Transmission Electron Microscopy (TEM) were used to characterize the nanoparticles. The synthetic route shows the formation of an anisotropic gold nanostructure consisting of spherical, triangles, hexagonals, and a low-yield rod with an aspect ratio ranging from 4.2 to 8.5. Curcumin-derived ctGNPs shows stability at different physiological conditions and better biocompatibility. Synthesized nanoparticles are found non-toxic towards eukaryotic cells but more effective against the cancer cell lines HeLa and MCF-7. The interaction of these synthesized nanoparticles with human telomeric G-quadruplex (GQ) DNA was studied using different physiochemical methods. The spectroscopic studies show that synthesized nanoparticles have stronger binding affinity towards telomeric GQ as compared to ds DNA through Van der Waals and H-bonding interactions. Thermodynamic interpretation reveals that the formation of the complex between the telomeric GQ and ctGNPs are enthalpy driven and entropy unfavourable process, resulting in motion freezing and, eventually, AuNP aggregation. Thus, our study shows a new approach to understand the interaction of telomeric G-quadruplexes with gold nanoparticles generated via the green route.

Продуктивные особенности коров голштинской породы разных генетических линий

Abstract. The purpose of the study is to investigate the productive features of Holstein cows of different genetic lines. Methods. The research was carried out in two breeding enterprises of the Sverdlovsk region on the livestock of Holstein cows. Groups of animals were formed by the method of pairs of analogues, taking into account the age of animals in lactation, physiological condition, live weight, lines of breeding bulls. The coefficient of milk content was estimated by the calculation method. The lactose, fat and protein content in milk was determined using a “Bentley 150” device. Somatic cells were determined on the “Somatos-M” device. Bacterial contamination was studied using a reductase assay. Prolactin was determined by enzyme immunoassay, cortisol by immunochemiluminescent method. Results. In terms of milk yield for 305 days of lactation and protein content in milk, Montwick Chieftain cows had an advantage with a significant difference between the groups (P < 0.05). The animals of the Reflection Sovering line were the best in terms of fat content, the difference between the groups was insignificant. The lowest content of somatic cells in milk and bacterial contamination was found in animals of the Reflection Sovering and Vis Back Idial lines. The best indicators of the leukocyte profile were determined in animals of the Vis Back Idial and Montwick Chieftain lines. In terms of glucose and total protein content in the blood, cows of the Montwick Chieftain line had an advantage, in terms of the amount of urea, animals of the Montwick Chieftain and Vis Back Idial line affiliation. The highest level of total lipids in the blood is typical for cows of the Reflection Sovering line. Scientific novelty. The research results indicate the relationship of biological characteristics and milk productivity with different lines of breeding bulls. The leukocyte profile of this herd of cows of different genetic lines in breeding enterprises of the Sverdlovsk region was studied for the first time. Practical significance. The results of the research can be used in breeding enterprises in order to increase the milk productivity of cows.