Perspectives on the Use of Hyaluronidase from the Red King Crab Hepatopancreas in Treating Filler Complications: In Vitro Investigations

The kinetics of the hydrolysis of hyaluronic acid (HA) of cosmetic fillers using thehomogenate of the red king crab hepatopancreas was studied for the first time. Turbidimetricanalysis of the reaction mixture revealed a bell–shaped time dependence of aggregation formation. The HA fillers were examined by atomic force microscopy (AFM) and it was found that they wererepresented by spherical–like structures. These structures were disrupted under the action of thehomogenate of the red king crab hepatopancreas. It was shown that the prepared homogenate hasthe activity which is similar to that observed in the commercially available hyaluronidase products.The preparation with hyaluronidase activity obtained from the red king crab hepatopancreas couldbe used as potentially safe product for treating filler complications.

Download Full-text

Perspectives on the Use of Hyaluronidase from the Red King Crab Hepatopancreas in Treating Filler Complications: In Vitro Investigations

10.20944/preprints201909.0001.v2 ◽

2019 ◽

Author(s):

Tatyana Ponomareva ◽

Dmitrii Sliadovskii ◽

Maria Timchenko ◽

Maxim Molchanov ◽

Alexander Timchenko ◽

...

Keyword(s):

Red King Crab ◽

King Crab ◽

Force Microscopy ◽

Atomic Force ◽

Safe Product ◽

Crab Hepatopancreas ◽

Aggregation Formation ◽

Turbidimetric Analysis ◽

Filler Complications

This study focused on hydrolysis of cosmetic fillers hyaluronic acid (HA) and kinetics of the HA hydrolysis using the homogenate of the red king crab hepatopancreas. Turbidimetric analysis of the reaction mixture revealed a bell-shaped time dependence of aggregation formation. It was shown that the obtained homogenate has the similar activity to the commercially available hyaluronidase. The atomic force microscopy (AFM) examination found that the HA fillers were represented by spherical-like structures. These structures were destroyed under the action of the homogenate of the red king crab hepatopancreas. NMR of the reaction mixture showed that HA degradation lasts for some days, but a maximum rate of the reaction is detected in the first hours of incubation. The preparation with hyaluronidase activity obtained from the red king crab hepatopancreas could be used as potentially safe product for treating filler complications.

Download Full-text

The effect of hepatopancreas homogenate of the Red king crab on HA-based filler

PeerJ ◽

10.7717/peerj.8579 ◽

2020 ◽

Vol 8 ◽

pp. e8579 ◽

Cited By ~ 1

Author(s):

Tatyana Ponomareva ◽

Dmitrii Sliadovskii ◽

Maria Timchenko ◽

Maxim Molchanov ◽

Alexander Timchenko ◽

...

Keyword(s):

Red King Crab ◽

King Crab ◽

Force Microscopy ◽

Atomic Force ◽

Hydrolysis Process ◽

Crab Hepatopancreas ◽

Turbidimetric Analysis ◽

Hydrolysis Of ◽

Microscopy Images ◽

Filler Complications

In this study, several methods were used to analyze the hydrolysis of hyaluronic acid (HA)-based cosmetic fillers by the hepatopancreas homogenate of the Red king crab. The results show that the homogenate and commercially available hyaluronidases have similar hydrolysis activities on the fillers. Atomic force microscopy images reveal that the HA fillers consist mainly of spherical-like particles, which are converted into filamentous structures as a result of hydrolysis by the Red king crab hepatopancreas homogenate. Turbidimetric analysis of the hydrolysis process shows that HA aggregation with acidic albumin exhibits a bell-shaped dependence on reaction time. Analysis of the hydrolysis process by nuclear magnetic resonance shows that HA degradation lasts several days. The maximum rate of the reaction is detected in the 1st h of incubation. The data confirm that the purified homogenate of the Red king crab hepatopancreas exerts hyaluronidase activity on HA-based cosmetic fillers; therefore, it may be considered as a potential therapeutic agent for treating filler complications.

Download Full-text

Exploring the action of RGDV-gemcitabine on tumor metastasis, tumor growth and possible action pathway

Scientific Reports ◽

10.1038/s41598-020-72824-8 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Xiaoyi Zhang ◽

Jinhuan Zhang ◽

Wenchao Liu ◽

Yaonan Wang ◽

Jianhui Wu ◽

...

Keyword(s):

Tumor Metastasis ◽

A549 Cells ◽

Force Microscopy ◽

Atomic Force ◽

Tnf Α ◽

First Time ◽

Inhibit Tumor Metastasis ◽

Necrosis Factor

Abstract The coupling of Arg-Gly-Asp-Val (RGDV) and gemcitabine led to a hypothesis that the conjugate (RGDV-gemcitabine) could inhibit tumor metastasis. To confirm this hypothesis the activities of RGDV-gemcitabine inhibiting tumor metastasis in vitro and in vivo were presented for the first time. AFM (atomic force microscopy) imaged that RGDV-gemcitabine was able to adhere onto the surface of serum-starved A549 cells, to block the extending of the pseudopodia. Thereby RGDV-gemcitabine was able to inhibit the invasion, migration and adhesion of serum-starved A549 cells in vitro. On C57BL/6 mouse model RGDV-gemcitabine dose dependently inhibited the metastasis of planted tumor towards the lung and the minimal dose was 0.084 µmol/kg/3 days. The decrease of serum TNF-α (tumor necrosis factor), IL-8 (interleukin-8), MMP-2 (matrix metalloprotein-2) and MMP-9 (matrix metalloprotein-9) of the treated C57BL/6 mice was correlated with the action pathway of RGDV-gemcitabine inhibiting the metastasis of the planted tumor towards lung.

Download Full-text

Mycobacterium tuberculosis and Paracoccidioides brasiliensis Formation and Treatment of Mixed Biofilm In Vitro

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.681131 ◽

2021 ◽

Vol 11 ◽

Author(s):

Kaila Petronila Medina-Alarcón ◽

Iara Pengo Tobias da Silva ◽

Giovana Garcia Ferin ◽

Marcelo A. Pereira-da-Silva ◽

Caroline Maria Marcos ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Paracoccidioides Brasiliensis ◽

Public Health Problem ◽

Extracellular Polysaccharide ◽

Force Microscopy ◽

Atomic Force ◽

High Degree ◽

First Time

Co-infection of Mycobacterium tuberculosis and Paracoccidioides brasiliensis, present in 20% in Latin America, is a public health problem due to a lack of adequate diagnosis. These microorganisms are capable of forming biofilms, mainly in immunocompromised patients, which can lead to death due to the lack of effective treatment for both diseases. The present research aims to show for the first time the formation of mixed biofilms of M. tuberculosis and P. brasiliensis (Pb18) in vitro, as well as to evaluate the action of 3’hydroxychalcone (3’chalc) -loaded nanoemulsion (NE) (NE3’chalc) against monospecies and mixed biofilms, the formation of mixed biofilms of M. tuberculosis H37Rv (ATCC 27294), 40Rv (clinical strains) and P. brasiliensis (Pb18) (ATCC 32069), and the first condition of formation (H37Rv +Pb18) and (40Rv + Pb18) and second condition of formation (Pb18 + H37Rv) with 45 days of total formation time under both conditions. The results of mixed biofilms (H37Rv + Pb18) and (40Rv + Pb18), showed an organized network of M. tuberculosis bacilli in which P. brasiliensis yeasts are connected with a highly extracellular polysaccharide matrix. The (Pb18 + H37Rv) showed a dense biofilm with an apparent predominance of P. brasiliensis and fragments of M. tuberculosis. PCR assays confirmed the presence of the microorganisms involved in this formation. The characterization of NE and NE3’chalc displayed sizes from 145.00 ± 1.05 and 151.25 ± 0.60, a polydispersity index (PDI) from 0.20± 0.01 to 0.16± 0.01, and zeta potential -58.20 ± 0.92 mV and -56.10 ± 0.71 mV, respectively. The atomic force microscopy (AFM) results showed lamellar structures characteristic of NE. The minimum inhibitory concentration (MIC) values of 3’hidroxychalcone (3’chalc) range from 0.97- 7.8 µg/mL and NE3’chalc from 0.24 - 3.9 µg/mL improved the antibacterial activity when compared with 3’chalc-free, no cytotoxicity. Antibiofilm assays proved the efficacy of 3’chalc-free incorporation in NE. These findings contribute to a greater understanding of the formation of M. tuberculosis and P. brasiliensis in the mixed biofilm. In addition, the findings present a new possible NE3’chalc treatment alternative for the mixed biofilms of these microorganisms, with a high degree of relevance due to the lack of other treatments for these comorbidities.

Download Full-text

Engineered Sumoylation-Deficient Prdx6 Mutant Protein-Loaded Nanoparticles Provide Increased Cellular Defense and Prevent Lens Opacity

Antioxidants ◽

10.3390/antiox10081245 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1245

Author(s):

Bhavana Chhunchha ◽

Eri Kubo ◽

Uday B. Kompella ◽

Dhirendra P. Singh

Keyword(s):

Protective Efficacy ◽

Plga Nanoparticles ◽

Lens Opacity ◽

Sustained Delivery ◽

Force Microscopy ◽

Cellular Defense ◽

Atomic Force ◽

Protein Dysfunction ◽

First Time

Aberrant Sumoylation-mediated protein dysfunction is involved in a variety of oxidative and aging pathologies. We previously reported that Sumoylation-deficient Prdx6K(lysine)122/142R(Arginine) linked to the TAT-transduction domain gained stability and protective efficacy. In the present study, we formulated wild-type TAT-HA-Prdx6WT and Sumoylation-deficient Prdx6-loaded poly-lactic-co-glycolic acid (PLGA) nanoparticles (NPs) to further enhance stability, protective activities, and sustained delivery. We found that in vitro and subconjuctival delivery of Sumoylation-deficient Prdx6-NPs provided a greater protection of lens epithelial cells (LECs) derived from human and Prdx6−/−-deficient mouse lenses against oxidative stress, and it also delayed the lens opacity in Shumiya cataract rats (SCRs) than TAT-HA-Prdx6WT-NPs. The encapsulation efficiencies of TAT-HA-Prdx6-NPs were ≈56%–62%. Dynamic light scattering (DLS) and atomic force microscopy (AFM) analyses showed that the NPs were spherical, with a size of 50–250 nm and a negative zeta potential (≈23 mV). TAT-HA-Prdx6 analog-NPs released bioactive TAT-HA-Prdx6 (6%–7%) within 24 h. Sumoylation-deficient TAT-HA-Prdx6-NPs provided 35% more protection by reducing the oxidative load of LECs exposed to H2O2 compared to TAT-HA-Prdx6WT-NPs. A subconjuctival delivery of TAT-HA-Prdx6 analog-NPs demonstrated that released TAT-HA-Prdx6K122/142R could reduce lens opacity by ≈60% in SCRs. Collectively, our results demonstrate for the first time that the subconjuctival delivery of Sumoylation-deficient Prdx6-NPs is efficiently cytoprotective and provide a proof of concept for potential use to delay cataract and oxidative-related pathobiology in general.

Download Full-text

BF0801, a novel adenine derivative, inhibits platelet activation via phosphodiesterase inhibition and P2Y12 antagonism

Thrombosis and Haemostasis ◽

10.1160/th10-05-0285 ◽

2010 ◽

Vol 104 (10) ◽

pp. 845-857 ◽

Cited By ~ 16

Author(s):

Si Zhang ◽

Liang Hu ◽

Hongguang Du ◽

Yan Guo ◽

Yan Zhang ◽

...

Keyword(s):

Platelet Aggregation ◽

Shape Change ◽

Atp Release ◽

Antiplatelet Drugs ◽

Force Microscopy ◽

Atomic Force ◽

Pde Inhibition ◽

First Time ◽

Adenine Derivative

SummaryThough antiplatelet drugs are proven beneficial to patients with coronary heart disease and stroke, more effective and safer antiplatelet drugs are still needed. In this study we report the antiplatelet effects and mechanism of BF0801, a novel adenine derivative. BF0801 dramatically inhibited platelet aggregation and ATP release induced by ADP, 2MeSADP, AYPGKF, SFLLRN or convulxin without affecting shape change in vitro. It also potentiated the inhibitory effects of adenosine-based P2Y12 antagonist AR-C69931MX or phosphodiesterase (PDE) inhibitor IBMX on platelet aggregation. The cAMP levels in both resting and forskolin-stimulated platelets were increased by BF0801 suggesting its PDE inhibitor activity, which is further confirmed by the concentration-dependent suppression of BF0801 on the native and recombinant PDE. Similar to AR-C69931MX, BF0801 drastically inhibited 2MeSADP-induced adenylyl cyclase inhibition in platelets indicating its P2Y12 antagonism activity, which is substantiated by the inhibition of BF0801 on the interaction between ADP and P2Y12 receptor expressed in CHO-K1 cells measured by atomic force microscopy. Moreover, we confirmed the antiplatelet effects of BF0801 using platelets from rats intravenously given BF0801. In summary, for the first time we developed a novel adenine derivative bearing dual activities of PDE inhibition and P2Y12 antagonism, which may have therapeutic advantage as a potential antithrombotic drug.

Download Full-text

Rapid, Refined, and Robust Method for Expression, Purification, and Characterization of Recombinant Human Amyloid-beta M1-42

10.26434/chemrxiv.7725002.v1 ◽

2019 ◽

Author(s):

Priya Prakash ◽

Travis Lantz ◽

Krupal P. Jethava ◽

Gaurav Chopra

Keyword(s):

Amyloid Beta ◽

Western Blots ◽

Force Microscopy ◽

E Coli ◽

Atomic Force ◽

Set Up ◽

Short Time

Amyloid plaques found in the brains of Alzheimer’s disease (AD) patients primarily consists of amyloid beta 1-42 (Ab42). Commercially, Ab42 is synthetized using peptide synthesizers. We describe a robust methodology for expression of recombinant human Ab(M1-42) in Rosetta(DE3)pLysS and BL21(DE3)pLysS competent E. coli with refined and rapid analytical purification techniques. The peptide is isolated and purified from the transformed cells using an optimized set-up for reverse-phase HPLC protocol, using commonly available C18 columns, yielding high amounts of peptide (~15-20 mg per 1 L culture) in a short time. The recombinant Ab(M1-42) forms characteristic aggregates similar to synthetic Ab42 aggregates as verified by western blots and atomic force microscopy to warrant future biological use. Our rapid, refined, and robust technique to purify human Ab(M1-42) can be used to synthesize chemical probes for several downstream in vitro and in vivo assays to facilitate AD research.

Download Full-text

Targeting Breast Cancer Cells with G4 PAMAM Dendrimers and Valproic Acid Derivative Complexes

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200423073812 ◽

2020 ◽

Vol 20 (15) ◽

pp. 1857-1872

Author(s):

Alberto M. Muñoz ◽

Manuel J. Fragoso-Vázquez ◽

Berenice P. Martel ◽

Alma Chávez-Blanco ◽

Alfonso Dueñas-González ◽

...

Keyword(s):

Valproic Acid ◽

Cell Lines ◽

Water Solubility ◽

Md Simulations ◽

Pamam Dendrimer ◽

Pamam Dendrimers ◽

Force Microscopy ◽

Micromolar Concentration ◽

Atomic Force

Background: Our research group has developed some Valproic Acid (VPA) derivatives employed as anti-proliferative compounds targeting the HDAC8 enzyme. However, some of these compounds are poorly soluble in water. Objective: Employed the four generations of Polyamidoamine (G4 PAMAM) dendrimers as drug carriers of these compounds to increase their water solubility for further in vitro evaluation. Methods: VPA derivatives were subjected to Docking and Molecular Dynamics (MD) simulations to evaluate their affinity on G4 PAMAM. Then, HPLC-UV/VIS, 1H NMR, MALDI-TOF and atomic force microscopy were employed to establish the formation of the drug-G4 PAMAM complexes. Results: The docking results showed that the amide groups of VPA derivatives make polar interactions with G4 PAMAM, whereas MD simulations corroborated the stability of the complexes. HPLC UV/VIS experiments showed an increase in the drug water solubility which was found to be directly proportional to the amount of G4 PAMAM. 1H NMR showed a disappearance of the proton amine group signals, correlating with docking results. MALDI-TOF and atomic force microscopy suggested the drug-G4 PAMAM dendrimer complexes formation. Discussion: In vitro studies showed that G4 PAMAM has toxicity in the micromolar concentration in MDAMB- 231, MCF7, and 3T3-L1 cell lines. VPA CF-G4 PAMAM dendrimer complex showed anti-proliferative properties in the micromolar concentration in MCF-7 and 3T3-L1, and in the milimolar concentration in MDAMB- 231, whereas VPA MF-G4 PAMAM dendrimer complex didn’t show effects on the three cell lines employed. Conclusion: These results demonstrate that G4 PAMAM dendrimers are capableof transporting poorly watersoluble aryl-VPA derivate compounds to increase its cytotoxic activity against neoplastic cell lines.

Download Full-text

High resolution nanoscale chemical analysis of bitumen surface microstructures

Scientific Reports ◽

10.1038/s41598-021-92835-3 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ayse N. Koyun ◽

Julia Zakel ◽

Sven Kayser ◽

Hartmut Stadler ◽

Frank N. Keutsch ◽

...

Keyword(s):

Secondary Ion Mass Spectrometry ◽

Force Microscopy ◽

Photo Oxidation ◽

Atomic Force ◽

Para Phase ◽

Key Sites ◽

Respective Surface ◽

Surface Domains ◽

Surface Microstructures ◽

First Time

AbstractSurface microstructures of bitumen are key sites in atmospheric photo-oxidation leading to changes in the mechanical properties and finally resulting in cracking and rutting of the material. Investigations at the nanoscale remain challenging. Conventional combination of optical microscopy and spectroscopy cannot resolve the submicrostructures due to the Abbe restriction. For the first time, we report here respective surface domains, namely catana, peri and para phases, correlated to distinct molecules using combinations of atomic force microscopy with infrared spectroscopy and with correlative time of flight—secondary ion mass spectrometry. Chemical heterogeneities on the surface lead to selective oxidation due to their varying susceptibility to photo-oxidation. It was found, that highly oxidized compounds, are preferentially situated in the para phase, which are mainly asphaltenes, emphasising their high oxidizability. This is an impressive example how chemical visualization allows elucidation of the submicrostructures and explains their response to reactive oxygen species from the atmosphere.

Download Full-text

Preparation and Characterization of Chitosan-Casein Phosphopeptides Biocomposite Coatings on the Medical Titanium Alloy

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.480-481.1065 ◽

2011 ◽

Vol 480-481 ◽

pp. 1065-1069

Author(s):

Bin Liu ◽

Lin Wang ◽

Yin Zhong Bu ◽

Sheng Rong Yang ◽

Jin Qing Wang

Keyword(s):

Photoelectron Spectroscopy ◽

Attenuated Total Reflectance ◽

Ti Alloy ◽

Force Microscopy ◽

Implant Materials ◽

Atomic Force ◽

Casein Phosphopeptides ◽

Potential Applications ◽

First Time

Titanium (Ti) and its alloys have been applied in orthopedics as one of the most popular biomedical metallic implant materials. In this work, to enhance the bioactivity, the surface of Ti alloy pre-modified by silane coupling agent and glutaraldehyde was covalently grafted with chitosan (CS) via biochemical multistep self-assembled method. Then, for the first time, the achieved surface was further immobilized with casein phosphopeptides (CPP), which are one group of bioactive peptides released from caseins in the digestive tract and can facilitate the calcium adsorption and usage, to form CS-CPP biocomposite coatings. The structure and composition of the fabricated coatings were characterized by X-ray photoelectron spectroscopy (XPS), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and atomic force microscopy (AFM). As the experimental results indicated, multi-step assembly was successfully performed, and the CS and CPP were assembled onto the Ti alloy surface orderly. It is anticipated that the Ti alloys modified by CS-CPP biocomposite coatings will find potential applications as implant materials in biomedical fields.

Download Full-text