Cellular Disruption Research Articles

Lipoma management with a minimally invasive 1,444 nm Nd:YAG laser technique.

Lipoma is the most common benign mesenchymal tumor that is composed of mature fat cells. Subdermal laser lipoma treatment may be recommended as an alternative to surgery for its removal. The purpose of the study was to investigate the efficacy of the 1,444 nm Nd:YAG laser subcutaneous intralesional application as a treatment option for lipoma. On 60 patients (37 women and 23 men) with lipomas localized above the muscle and lipomatosis in various regions, a subcutaneous, micro-pulsed 1,444 nm Nd:YAG laser procedure was executed. Before treatment, an ultrasound was performed and the lipomas were measured. The same lighting setup and photographic tools were used to take pictures of each patient. The lipoma reduced or completely disappeared in all cases at the last follow-up, and no infections, burns, skin lesions, episodes of severe bleeding, or other serious adverse effects were reported. The most common transient side effects were ecchymosis and edema. Partial lesion reduction refers to rare cases of lipomatosis in which the lipomas were so small that suction and accurate positioning of the capsular membrane contours were impossible. Lipoma treatment with a 1,444 nm Nd:YAG laser is a safe and effective minimally invasive procedure without risk of scarring. For cellular disruption, laser treatment is an effective and safe option.

An Optimised Monophasic Faecal Extraction Method for LC-MS Analysis and Its Application in Gastrointestinal Disease.

Liquid chromatography coupled with mass spectrometry (LC-MS) metabolomic approaches are widely used to investigate underlying pathogenesis of gastrointestinal disease and mechanism of action of treatments. However, there is an unmet requirement to assess faecal metabolite extraction methods for large-scale metabolomics studies. Current methods often rely on biphasic extractions using harmful halogenated solvents, making automation and large-scale studies challenging. The present study reports an optimised monophasic faecal extraction protocol that is suitable for untargeted and targeted LC-MS analyses. The impact of several experimental parameters, including sample weight, extraction solvent, cellular disruption method, and sample-to-solvent ratio, were investigated. It is suggested that a 50 mg freeze-dried faecal sample should be used in a methanol extraction (1:20) using bead beating as the means of cell disruption. This is revealed by a significant increase in number of metabolites detected, improved signal intensity, and wide metabolic coverage given by each of the above extraction parameters. Finally, we addressed the applicability of the method on faecal samples from patients with Crohn's disease (CD) and coeliac disease (CoD), two distinct chronic gastrointestinal diseases involving metabolic perturbations. Untargeted and targeted metabolomic analysis demonstrated the ability of the developed method to detect and stratify metabolites extracted from patient groups and healthy controls (HC), highlighting characteristic changes in the faecal metabolome according to disease. The method developed is, therefore, suitable for the analysis of patients with gastrointestinal disease and can be used to detect and distinguish differences in the metabolomes of CD, CoD, and HC.

Cysteamine capped silver nanoclusters: A potential antimicrobial agent for antibiotic-resistant bacteria

The world has witnessed a phenomenal increase in the frequent use of antibiotics and antimicrobial resistance for pathogens, which now are a great challenge for scientists and researchers. Consequently, there is an urgent need for novel antibacterial agents to combat pathogenic bacterial infections. This work sheds light on the relatively scarce application of silver nanoclusters (AgNCs) for the development of novel antimicrobial agents for antibiotic-resistant bacteria. We have prepared noble metal NCs (comprising 5 Ag atoms per cluster) by using a small scaffold,viz. cysteamine (CyA). Various spectroscopic and microscopic techniques were used to characterize the synthesized luminescent AgNCs. The composition of the NCs has been substantiated by mass spectral analyses which were well-supported by the Jellium model. The as-prepared NCs display efficacious antibacterial activity against common bacteria (both Gram-negative and Gram-positive) and also demonstrate promising bactericidal effects against ampicillin and kanamycin-resistant transformed strains of Escherichia coli bacteria. From in-depth systematic studies, the bactericidal activity was found due to the generation of reactive oxygen species (ROS) and cellular membrane disruption in the presence of AgNCs which resulted in the leakage of intracellular components, like proteins. Our study suggests that cysteamine-capped AgNCs might be used as effective antimicrobial agents in eliminating antibiotic-resistant bacteria for the development of a new kind of nanobiotics to combat multidrug-resistant deadly bacteria.

THROMBOCYTOPENIA, HYPOGAMMAGLOBULINEMIA, AND POOR VACCINE RESPONSE IN A PATIENT WITH TAKENOUCHI-KOSAKI SYNDROME

<h3>Introduction</h3> Takenouchi-Kosaki Syndrome (TKS) is a rare congenital developmental disorder characterized by thrombocytopenia, facial abnormalities, and varied multisystemic anomalies including immunodeficiency. Heterozygous deficiency of CDC42, an intracellular signaling GTPase, has been found in all TKS patients described to date. The immunological phenotypes of TKS patients are heterogeneous: recurrent infections have been noted in some patient series but with variable penetrance. Herein we describe a case of TKS causing hypogammaglobulinemia, B-cell lymphopenia, and poor vaccine response. <h3>Case Description</h3> A fifteen-year-old girl with history of developmental delay, multiple congenital defects, and lymphedema was referred to the immunology clinic for evaluation of recurrent ear infections that began at 3 months of age requiring tympanostomy tube placement at 18 months. Her physical exam was notable for abnormal facies, scoliosis, strabismus, abnormal thumbs, and unilateral lymphedema. Her laboratory evaluation revealed macrothrombocytopenia, mild leukopenia with low T-cells and markedly low B-cells by flow cytometry, and notably low IgA (11.7, RR 69 – 309) and IgG (423, RR 613 – 1295). She had a poor response to a full course of PCV7 with PPSV23 booster (9 of 23 > 1.3). Whole exome sequencing identified a heterozygous p.Y64C mutation in CDC42 consistent with TKS. <h3>Discussion</h3> The Y64C mutation in CDC42 is a TKS Group I lesion causing ineffective intracellular signaling, resulting in <i>in vitro</i> defects including abnormal cellular proliferation and disruption of cell polarity. Actin remodeling disorders such as Wiskott-Aldrich Syndrome and TKS should be considered in patients with immunodeficiency and congenital thrombocytopenia.

Reversible and Irreversible Effects of Electroporation on Contractility and Calcium Homeostasis in Isolated Cardiac Ventricular Myocytes.

Irreversible electroporation is an energy form utilizing high-voltage pulsed electric field, leading to cellular homeostasis disruption and cell death. Recently, irreversible electroporation has shown promising results for the treatment of cardiac arrhythmias. However, reversible and irreversible effects of pulsed electric field on cardiac myocytes remain poorly understood. Here, we evaluated the influence of a monophasic single electric pulse (EP) on the contractility, Ca2+ homeostasis and recovery of cardiac myocytes. Isolated rat left ventricular myocytes were electroporated using single monophasic EP of different durations and voltages. Sarcomere length and intracellular Ca2+ were simultaneously monitored for up to 20 minutes after EP application in Fura-2 loaded left ventricular myocytes. Lethal voltage thresholds were determined using 100 µs and 10 ms pulses and by discriminating cell orientation with respect to the electric field. Electroporation led to an immediate increase in intracellular Ca2+ which was dependent upon the voltage delivered to the cell. Intermediate-voltage EP (140 V, 100 µs) increased sarcomere shortening, Ca2+ transient amplitude, and diastolic Ca2+ level measured 1 minute post-EP. Although sarcomere shortening returned to pre-EP level within 5 minutes, Ca2+ transient amplitude decreased further below pre-EP level and diastolic Ca2+ level remained elevated within 20 minutes post-EP. Spontaneous contractions were observed after sublethal EP application but their frequency decreased progressively within 20 minutes. Lethal EP voltage threshold was lower in myocytes oriented perpendicular than parallel to the electric field using 100 µs pulses while an opposite effect was found using 10 ms pulses. Sublethal EP affected rat left ventricular myocytes contractility and disrupted Ca2+ homeostasis as a function of the EP voltage. Moreover, EP-induced lethality was preceded by a large increase in intracellular Ca2+ and was dependent upon the EP duration, amplitude and left ventricular myocytes orientation with respect to the electric field. These findings provide new insights into the effect of pulsed electric field on cardiac myocytes.

Poststroke Seizure and Epilepsy: A Review of Incidence, Risk Factors, Diagnosis, Pathophysiology, and Pharmacological Therapies

Stroke is the most common cause of epilepsy and ultimately leads to a decrease in the quality of life of those affected. Ischemic and hemorrhagic strokes can both lead to poststroke epilepsy (PSE). Significant risk factors for PSE include age < 65age less than 65 years, stroke severity measured by the National Institutes of Health Stroke Scale (NIHSS), cortical involvement, and genetic factors such as TRPM6 polymorphism. The diagnosis of PSE is made by using imaging modalities, blood biomarkers, and prognostic criteria. Electroencephalography (EEG) is currently the gold standard to diagnose PSE, while new combinations of modalities are being tested to increase diagnostic specificity. This literature review uncovers a newly found mechanism for the pathology of poststroke epilepsy. The pathogenesis of early-onset and late-onset is characterized by sequelae of neuronal cellular hypoxia and disruption of the blood-brain barrier, respectively. Interleukin-6 is responsible for increasing the activity of glial cells, causing gliosis and hyperexcitability of neurons. Epinephrine, high-mobility group protein B1, downregulation of CD32, and upregulation of HLA-DR impact the pathology of poststroke epilepsy by inhibiting the normal neuronal immune response. Decreased levels of neuropeptide Y, a neurotransmitter, act through multiple unique mechanisms, such as inhibiting intracellular Ca2+ accumulation and acting as an anti-inflammatory, also implemented in the worsening progression of poststroke epilepsy. Additionally, CA1 hippocampal resonant neurons that increase theta oscillation are associated with poststroke epilepsy. Hypertensive small vessel disease may also have an implication in the temporal lobe epilepsy by causing occult microinfarctions. Furthermore, this review highlights the potential use of statins as primary prophylaxis against PSE, with multiple studies demonstrating a reduction in incidence using statins alone, statins in combination with antiepileptic drugs (AEDs), and statins with aspirin. The evidence strongly suggests that the second generation AEDs are a superior treatment method for PSE. Data from numerous studies demonstrate their relative lack of significant drug interactions, increased tolerability, and potential superiority in maintaining seizure-free status.

Enzymatic Hydrolysis of Pacific White Shrimp Residue (Litopenaeus vannamei) with Ultrasound Aid

ABSTRACT Several methods are used to recover compounds from these residues. Among them, ultrasound extraction has gained prominence due to its cellular disruption power. This work aimed to verify the action of ultrasound in increasing the yield of protein hydrolysate extraction from Pacific white shrimp (Litopenaeus vannamei) by-products. Proximal composition, hydrolysis degree, total content of carotenoids, and antioxidant activity (DPPH, ABTS, and reducing power) of the obtained hydrolysate was determined. The study found that the use of ultrasound concentrated more carotenoids, containing high antioxidant activity, increased the degree of hydrolysis, and accelerated the hydrolytic reaction rate.

Antitumoral effect of novel synthetic 8-hydroxy-2-((4-nitrophenyl)thio)naphthalene-1,4-dione (CNN16) via ROS-mediated DNA damage, apoptosis and anti-migratory effect in colon cancer cell line

Colorectal cancer (CRC) is estimated as the third most incident cancer and second in mortality worldwide. Moreover, CRC metastasis reduces patients' survival rates. Thus, the study and identification of new compounds with anticancer activity selectively to tumor cells are encouraged in the CRC treatment. Naphtoquinones are compounds with several pharmacologic activities, including antitumoral properties. Therefore, this study aimed to investigate the anticancer mechanism of synthetic 8-Hydroxy-2-(P-Nitrothiophenol)-1,4-Naphthoquinone (CNN16) in colon cancer cell line HCT-116. CNN16 showed an IC50 of 5.32 μM in HCT-116, and 9.36, 10.77, and 24.57 μM in the non-cancerous cells MRC-5, MNP-01, and PMBC, respectively, evaluated by the MTT assay. CNN16 showed an anticlonogenic effect in HCT-116 and induced cell fragmentation identified by flow cytometry analysis. Furthermore, we observed that CNN16 presented genotoxicity and induces reactive oxygen species (ROS) after 3 h of treatment visualized by alkaline comet assay and DCFH-DA dye fluorescence, respectively. Furthermore, CNN16 caused cellular membrane disruption, reduction in the mitochondrial membrane polarization, and the presence of apoptotic bodies and chromatin condensation was visualized by differential stained (HO/FD/PI) in fluorescent microscopy along with PARP1, TP53, BCL-2, and BAX analyzed by RT-qPCR. Results also evidenced inhibition in the migratory process analyzed by wound healing assay. Therefore, CNN16 can be considered as a potential new leader molecule for CRC treatment, although further studies are still necessary to comprehend the effects of CNN16 in in vivo models to evaluate the anti-migratory effect, and toxicology and assure compound safety and selectively.

HMGA2 mediates Cr (VI)-induced metabolic reprogramming through binding to mitochondrial D-Loop region.

Hexavalent chromium [Cr (VI)] exists environmentally and occupationally. It has been shown to pose a carcinogenic hazard in certain occupations. This study was to investigate the role of high mobility group A2 (HMGA2) in Cr (VI)-induced metabolism reprogramming from oxidative phosphorylation (OXPHOS) to glycolysis in A549 and HELF cells. First, knockdown of HMGA2 by siHMGA2 significantly attenuated Cr (VI)-reduced expression of OXPHOS-related proteins (COX IV and ND1) and mitochondrial mass, indicating that HMGA2 was involved in Cr (VI)-reduced OXPHOS. Overexpression of HMGA2 by transfection of HMGA2-DNA plasmids reduced the expression of COX IV, ND1 and mitochondrial mass, suggesting the negative role of HMGA2 in OXPHOS. Secondly, both CCCP, the inhibitor of mitochondrial function, and the ER stress inhibitor, 4-phenylbutyric acid (4-PBA), decreased the level of HMGA2, indicating that the interaction of mitochondrial dysfunction and ER stress resulted in Cr (VI)-induced HMGA2 expression. Further study demonstrated that ER stress/HMGA2 axis mediated the metabolism rewiring from OXPHOS to aerobic glycolysis. Notably, Cr (VI) induced the accumulation of HMGA2 proteins in mitochondria and ChIP assay demonstrated that HMGA2 proteins could bind to D-loop region of mitochondrial DNA (mtDNA), which provided the proof for HMGA2-modulating OXPHOS. Taken together, our results suggested that the interaction of mitochondria and ER stress-enhanced HMGA2 played an important role in Cr (VI)-induced metabolic reprogramming from OXPHOS to glycolysis by binding directly to D-loop region of mtDNA. This work informs on the potential mode of action for Cr (VI)-induced tumors and builds on growing evidence regarding the contribution of cellular metabolic disruption contributing to carcinogenicity.

Unravelling soluble immune checkpoints in chronic lymphocytic leukemia: Physiological immunomodulators or immune dysfunction.

Chronic lymphocytic leukemia (CLL) is a lymphoid neoplasm characterized by the accumulation of mature B cells. The diagnosis is established by the detection of monoclonal B lymphocytes in peripheral blood, even in early stages [monoclonal B-cell lymphocytosis (MBLhi)], and its clinical course is highly heterogeneous. In fact, there are well-characterized multiple prognostic factors that are also related to the observed genetic heterogenicity, such as immunoglobulin heavy chain variable region (IGHV) mutational status, del17p, and TP53 mutations, among others. Moreover, a dysregulation of the immune system (innate and adaptive immunity) has been observed in CLL patients, with strong impact on immune surveillance and consequently on the onset, evolution, and therapy response. In addition, the tumor microenvironment is highly complex and heterogeneous (i.e., matrix, fibroblast, endothelial cells, and immune cells), playing a critical role in the evolution of CLL. In this study, a quantitative profile of 103 proteins (cytokines, chemokines, growth/regulatory factors, immune checkpoints, and soluble receptors) in 67 serum samples (57 CLL and 10 MBLhi) has been systematically evaluated. Also, differential profiles of soluble immune factors that discriminate between MBLhi and CLL (sCD47, sCD27, sTIMD-4, sIL-2R, and sULBP-1), disease progression (sCD48, sCD27, sArginase-1, sLAG-3, IL-4, and sIL-2R), or among profiles correlated with other prognostic factors, such as IGHV mutational status (CXCL11/I-TAC, CXCL10/IP-10, sHEVM, and sLAG-3), were deciphered. These results pave the way to explore the role of soluble immune checkpoints as a promising source of biomarkers in CLL, to provide novel insights into the immune suppression process and/or dysfunction, mostly on T cells, in combination with cellular balance disruption and microenvironment polarization leading to tumor escape.

Ecofriendly phytosynthesized zirconium oxide nanoparticles as antibiofilm and quorum quenching agents against Acinetobacter baumannii

The worldwide increase of multi-drug resistance has directed the researchers to focus on ecofriendly ways of nanoparticles synthesis with effective antivirulence properties. Here, we report the antibiofilm and quorum quenching (QQ) potential of zirconium oxide nanoparticles (ZrO2 NPs) synthesized from aqueous ginger extract against multi-drug resistant (MDR) Acinetobacter baumannii. The results indicated that ZrO2 NPs were of tetragonal shape with average diameter of 16 nm. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values for A. baumannii were 15.6 and 62.5 µg/ml, respectively, as revealed by broth microdilution assay. Exposure of bacterial cells to ZrO2 NPs resulted in reactive oxygen species (ROS) generation which in turn led to cellular membrane disruption as observed by an increase in leakage of cellular contents, such as proteins, sugars, and DNA. The antibiofilm activity was evaluated by microtiter plate assay and the results revealed that the percentage inhibition of biofilm was found to be 14.3–80.6%. ZrO2 NPs also obstructed the chemical composition of biofilms matrix by reducing the proteins and carbohydrate contents. Molecular docking studies of ZrO2 NPs with four proteins (2NAZ, 4HKG, 5D6H, and 5HM6) involved in biofilm formation of A. baumannii revealed the interaction of zirconium with target proteins. These findings suggested the in vitro efficacy of phytosynthesized ZrO2 NPs as antibiofilm and QQ agents that can be exploited in the development of alternative therapeutic options against MDR A. baumannii.

Novel Triterpenic Acid-Benzotriazole Esters Act as Pro-Apoptotic Antimelanoma Agents.

Pentacyclic triterpenes, such as betulinic, ursolic, and oleanolic acids are efficient and selective anticancer agents whose underlying mechanisms of action have been widely investigated. The introduction of N-bearing heterocycles (e.g., triazoles) into the structures of natural compounds (particularly pentacyclic triterpenes) has yielded semisynthetic derivatives with increased antiproliferative potential as opposed to unmodified starting compounds. In this work, we report the synthesis and biological assessment of benzotriazole esters of betulinic acid (BA), oleanolic acid (OA), and ursolic acid (UA) (compounds 1–3). The esters were obtained in moderate yields (28–42%). All three compounds showed dose-dependent reductions in cell viability against A375 melanoma cells and no cytotoxic effects against healthy human keratinocytes. The morphology analysis of treated cells showed characteristic apoptotic changes consisting of nuclear shrinkage, condensation, fragmentation, and cellular membrane disruption. rtPCR analysis reinforced the proapoptotic evidence, showing a reduction in anti-apoptotic Bcl-2 expression and upregulation of the pro-apoptotic Bax. High-resolution respirometry studies showed that all three compounds were able to significantly inhibit mitochondrial function. Molecular docking showed that compounds 1–3 showed an increase in binding affinity against Bcl-2 as opposed to BA, OA, and UA and similar binding patterns compared to known Bcl-2 inhibitors.

Abstract 007: Hypertension-induced Neurovascular Dysfunction At Single-cell Resolution

Hypertension (HTN) disrupts vital neurovascular control mechanisms, thereby increasing the brain’s susceptibility to vascular insufficiency, white matter lesions, and cognitive impairment. Yet, the distinct vascular, neuronal, and glial cell types targeted by HTN, as well as the ensuing cellular network disruption driving the neurovascular and cognitive deficits remain undefined. Here we sought to uncover transcriptomic changes in neurons and vascular cells using unbiased, single-cell RNA sequencing on the neocortex of 10-week old C57BL/6 male mice with angiotensin II (AngII) HTN. Vehicle or AngII (600ng/kg/min s.c.) were administered for 3 days, when blood-brain barrier (BBB) permeability start to increase, or 42 days, when neurovascular and cognitive dysfunction are fully developed (n=3/group). We analyzed 39,451 single-cell transcriptomes comprising 26 cell types. Surprisingly, 3 days of AngII induced significantly greater transcriptional changes in venular ECs compared to arteriolar ECs (EdgeR; pval&lt; 0.05, logFC&gt; 2), supporting the notion that venular ECs are uniquely sensitive to the early effects of HTN (Hypertension 76:795, 2020). Gene ontology analysis of differentially expressed genes prominently implicated altered venular immune signaling, BBB dysfunction, and, notably, a secretory phenotype characteristic of senescence (pval &lt;0.05). Moreover, unbiased ligand-receptor interaction analysis (CellChat) demonstrated that senescent venular ECs strongly communicate with oligodendrocyte precursors, and NPY-expressing interneurons, pointing to a previously unrecognized early disruption in the oligo-vascular niche, essential for maintaining white matter integrity, and neuronal network stability. Furthermore, at 42 days of AngII we observed an overrepresentation of aging and neurodegeneration-linked genes in oligos and NPY interneurons, relating to myelin disruption, synaptic dysfunction, and metabolic dysregulation. The data reveal a novel endothelial-oligo-interneuron crosstalk and transcriptomic alterations underlying the impact of HTN on the brain. Future studies will establish how these transcriptomic changes are linked to neurovascular dysfunction, white matter damage and cognitive impairment.

Current Challenges in Image-Guided Magnetic Hyperthermia Therapy for Liver Cancer.

For patients diagnosed with advanced and unresectable hepatocellular carcinoma (HCC), liver transplantation remains the best option to extend life. Challenges with organ supply often preclude liver transplantation, making palliative non-surgical options the default front-line treatments for many patients. Even with imaging guidance, success following treatment remains inconsistent and below expectations, so new approaches are needed. Imaging-guided thermal therapy interventions have emerged as attractive procedures that offer individualized tumor targeting with the potential for the selective targeting of tumor nodules without impairing liver function. Furthermore, imaging-guided thermal therapy with added standard-of-care chemotherapies targeted to the liver tumor can directly reduce the overall dose and limit toxicities commonly seen with systemic administration. Effectiveness of non-ablative thermal therapy (hyperthermia) depends on the achieved thermal dose, defined as time-at-temperature, and leads to molecular dysfunction, cellular disruption, and eventual tissue destruction with vascular collapse. Hyperthermia therapy requires controlled heat transfer to the target either by in situ generation of the energy or its on-target conversion from an external radiative source. Magnetic hyperthermia (MHT) is a nanotechnology-based thermal therapy that exploits energy dissipation (heat) from the forced magnetic hysteresis of a magnetic colloid. MHT with magnetic nanoparticles (MNPs) and alternating magnetic fields (AMFs) requires the targeted deposition of MNPs into the tumor, followed by exposure of the region to an AMF. Emerging modalities such as magnetic particle imaging (MPI) offer additional prospects to develop fully integrated (theranostic) systems that are capable of providing diagnostic imaging, treatment planning, therapy execution, and post-treatment follow-up on a single platform. In this review, we focus on recent advances in image-guided MHT applications specific to liver cancer

Synthetic oleanane triterpenoid derivative CDDO-Me disrupts cellular bioenergetics to suppress pancreatic ductal adenocarcinoma via targeting SLC1A5.

To investigate the potential antitumor activity of synthetic triterpenoid, methyl-2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oate(CDDO-Me) in pancreatic ductal adenocarcinoma (PDAC), MTT cytotoxicity assay, and xenograft nude mice assay were performed to evaluate tumor growth in vitro and in vivo. Seahorse XFe96 bioenergetics analyzer was applied to determine aerobic glycolysis and mitochondrial respiration. Western blot and quantitative reverse transcription-polymerase chain reactions areused to detect protein and messenger RNA transcripts of SLC1A5 and metabolic enzymes. We confirmed the strong antitumor activity of CDDO-Me in suppressing PDAC growth. Mechanistically, we demonstrated CDDO-Me induced mitochondrial respiration and aerobic glycolysis dysfunction. We also verified CDDO-Me downregulated glutamine transporter SLC1A5, resulting in excessive reactive oxygen species(ROS) levels that suppressed tumor growth. Moreover, we confirmed thatSLC1A5 depletion reduced the ratio of glutathione/oxidized glutathione.We also found CDDO-Me could inhibit N-linked glycosylation of SLC1A5, which promotes protease-mediated degradation. Finally, we confirmed SLC1A5 was significantly overexpressed in PDAC and closely correlated with the poor prognosis of PDAC patients. Our work uncovers CDDO-Me is effective at suppressing PDAC cell growth in vitro and in vivo and illuminates CDDO-Me caused excessive ROS and cellular bioenergetics disruption which contributed to CDDO-Me inhibited PDAC growth. Our data highlights CDDO-Me could be considered a potential compound for PDAC therapy, and SLC1A5 could be a novel biomarker for PDAC patients.

Liposomes encapsulating novel antimicrobial peptide Omiganan: Characterization and its pharmacodynamic evaluation in atopic dermatitis and psoriasis mice model

Omiganan is a novel 12 amino acid synthetic cationic peptide from the cathelicidin family. Omiganan possesses antimicrobial action against a wide range of microbes, including gram-positive and gram-negative bacteria and fungi. Omiganan mainly acts by depolarizing the cytoplasmic membrane, resulting in cellular disruption and death. Apart from its antimicrobial effect, Omiganan also has anti-inflammatory activity. The present investigation aimed to evaluate and compare the efficacy of Omiganan liposomal gel with conventional formulations (Omiganan gel and lotion) in atopic dermatitis (AD) and psoriasis mice animal models. Liposomes encapsulating Omiganan were prepared using the reverse-phase evaporation technique and incorporated into Carbopol 934P gel. The optimized Omiganan liposomes were then characterized for various physicochemical parameters such as vesicle size, shape and surface morphology, zeta-potential, rheological parameters, in-vitro drug release, ex-vivo skin permeation/deposition, in-vitro antimicrobial activity, proteolytic stability, and cellular toxicity and uptake studies. Liposomes exhibited 72 % encapsulation with 7.8 % loading efficacy, a vesicle size, and zeta potential of 120 nm and − 17.2 mv, respectively. Moreover, Omiganan liposomal gel demonstrated controlled release and a better permeation profile than conventional formulations. A substantial reduction in levels of pro-inflammatory cytokines and improvement in AD and psoriatic lesions were achieved by Omiganan liposomal gel compared to Omiganan gel and lotion-based formulations. The present study confirms that Omiganan liposomal formulation can be an effective, safe, and novel alternative treatment approach in atopic dermatitis and psoriasis.

Catechol-Based Polymers with High Efficacy in Cytosolic Protein Delivery

Catechol-Based Polymers with High Efficacy in Cytosolic Protein Delivery

O-191 Endometriosis is associated with a lowered cumulative live birth rate by alters the macrophage polarization and the cytokine concentrations in follicular fluid

Abstract Study question The impact of endometriosis on the outcomes of ART is still ambiguous and the mechanism by which endometriosis impacts fertility has not been fully elucidated Summary answer Endometriosis was associated with low CLBR in IVF, which might be due to the change of follicular microenvironment and compromising the quality of embryos. What is known already Despite the great advantages of IVF treatment in endometriosis-linked infertility, the impact of endometriosis on outcomes of infertility management with ART is controversial and few studies focused on the association between endometriosis and CLBR. Numerous mechanisms have been proposed in an effort to delineate the multifaceted pathophysiology that induces impairment of reproductive dynamics in patients with endometriosis. Reactive oxygen species, dysregulation of the immune system and cellular architectural disruption constitute the crucial mechanisms that detrimentally affect oocyte and embryo developmental potential. Study design, size, duration This study retrospectively included 433 patients with endometriosis and 1299 patients with tuber factor infertility to evaluate the impact of endometriosis on IVF pregnancy outcome between January 2016 and December 2018.The basic study prospectively recruited 30 patients with ovarian endometriosis and 35 controls with tubal factor infertility to analyze polarization stage of macrophages in their follicular fluid, and another 20 females with ovarian endometriosis and 30 controls to detect cytokines in their FF. Participants/materials, setting, methods Data on all women undergoing fresh or frozen IVF treatment cycles were analysed to compare the CLBR between endometriosis and tuber factor infertility patients. A cytometry panel of 4 antibodies (CD45, CD3, CD80, CD163) was designed to enumerate the numbers of MI and MII macrophages from the cell sediment from FF samples. Quantibody® array was utilized to determine the concentration of 10 cytokines in FF, including IFN-γ, IL-1α, IL-1β, IL-10,IL-13,IL-4, IL-6, IL-8, MCP-1 and TNF-α. Main results and the role of chance The results showed that patients with endometriosis were associated with noticeably fewer retrievable oocytes, a lower oocyte maturity rate, decreased numbers of available and high-quality embryos (all p &amp;lt; 0.001) in comparison with the control group. The clinical pregnancy and live birth rate of the endometriosis group were lower in the frozen-thawed embryo transfer cycles (p = 0.028 and p = 0.008, respectively), which leading to a declined cumulative live birth rate (CLBR) (p = 0.001). Logistic regression analysis indicated a close association between endometriosis and low CLBR (p = 0.002). Furthermore, the numbers of type I and type II macrophages in follicular fluid (FF) of patients with ovarian endometriosis were significantly increased compared with the control group (p &amp;lt; 0.001). The expressions of IL-1α, IL-1β, TNF-α, IL-6, IL-13, and IL-10 in FF were higher in endometrioma group(p &amp;lt; 0.05). The increased expression of cytokines was negatively correlated with embryo outcomes, including the numbers of total oocytes retrieved, mature oocytes, and fertilized oocytes, and the numbers of high-quality blastocysts and embryos. Additionally, IL-6 and IL-8 were positively correlated with AMH, rate of available blastocysts, number of blastocysts formed and available blastocysts, but were negatively associated with rate of MII oocytes. Limitations, reasons for caution The present study being of a retrospective and monocentric study, may not come to a very convincing conclusion, and some unknown biases might still exist due to possible underestimation of some confounders. Wider implications of the findings Collectively, our results indicate that endometriosis does adversely affect pregnancy outcomes of ART, as women with endometriosis produce a lower quantity of oocytes and embryos. Moreover, endometriosis might play a role in oocyte or embryo outcomes via regulating the production of cytokines or the number of immune cells in FF. Trial registration number no

What is the pro-inflammatory dietary model associated with?

Introduction. Cellular potential disruption leads to several polyplatoligias due to increased systemic inflammation, as a consequence of the visualized multivariate appearance. Objective. To highlight the relevance of the pathomechanism of oxidative stress and oxidative potential about changes in metabolic mechanisms at the cellular level, predisposing to polypatoligia and the development of destructive transformations visualized in outward appearance targeted by a pro-inflammatory nutritional model. Material and methods. A sample of 226 subjects was selected by random selection of respondents. The author's questionnaire concerned metric data, level of physical exercise, diagnosed disease entity, inflammatory changes within the body dermis, and level of frequency of consumption of selected food groups. The obtained results were verified by statistical analysis with a significance level of α &lt;0.05. Results. The subject was characterized by a diagnosed multimorbidity at the level of (approximately 20%) about the individual clinical course of the respondent. The anthropometric parameter BMI without sex division (due to predominance of female subjects) among 55% of the whole group was within the normal range, constituting 24.4 kg/m2, showed a significant correlation directed at the pathomechanism of multimorbidity at the level of 78% in physical appearance. Conclusions. The pathomechanism of developing polyposis is significantly associated with the appearance of changes within the tissues translated into the shell system and anthropometric measurements related to the value of body mass index. Multivariable within the common coat is significantly dependent on the implemented phytotherapeutics - bioactive components derived from oregano, to obtain high antioxidant potential.

Surface-enhanced Raman spectroscopy enabled evaluation of bacterial inactivation

An improved understanding of bacterial inactivation mechanisms will provide useful insights for infectious disease control and prevention. We evaluated bacterial response to several inactivation methods using surface-enhanced Raman spectroscopy (SERS). The results indicate that changes in the SERS signal are highly related to cellular disruption and that cellular changes arising after cell inactivation cannot be ignored. The membrane integrity of heat and the combination of UV254 and free chlorine (UV254/chlorine) treated Pseudomonas syringae (P. syringae) cells were severely disrupted, leading to significantly increased peak intensities. Conversely, ethanol treated bacteria exhibited intact cell morphologies and the SERS spectra remained virtually unchanged. On the basis of time dependent SERS signals, we extracted dominant SERS patterns. Peaks related to nucleic acids accounted for the main changes observed during heat, UV254, and UV254/chlorine treatment, likely due to their outward diffusion from the cell cytoplasm. For free chlorine treated P. syringae, carbohydrates and proteins on the cell membrane were denatured or lost, resulting in a decrease in related peak intensities. The nucleobases were likely oxidized when treated with UV254 and chlorine, thus leading to shifts in the related peaks. The generality of the method was verified using two additional bacterial strains: Escherichia coli and Bacillus subtilis as well as in different water matrices. The results suggest that SERS spectral analysis is a promising means to examine bacterial stress response at the molecular level and has applicability in diverse environmental implementations.