Confocal Laser Fluorescence Microscopy Research Articles

Transparent silicone-free shampoo containing proteins with strong hair conditioning properties.

With the constant upgrading of healthcare concepts, silicone-free hair products have attracted more attention among consumers. In the present study, transparent silicone-free shampoo containing proteins was successfully fabricated by mixing mild non-sulphate surfactants, compound cationic conditioners, natural silicone oil substitutes, protein conditioners, thickeners, and other auxiliary ingredients. The effects of the type of surfactants, hair conditioners and thickeners, the type and content of proteins, and the mass ratio of compound proteins on the hair foaming performance, hair grooming performance, and penetration performance were investigated. The basic formulation framework for transparent silicone-free shampoo was established at first. Then, various hydrolyzed proteins were further added to the basic formulation in the form of single use or compound use to prepare transparent silicone-free shampoo containing different proteins. The morphology of hair samples and penetration of protein in hair were evaluated with a scanning electron microscope and laser confocal fluorescence microscopy, respectively. And the hair grooming performance was also determined by a dynamic combing tester. The compound proteins of Croquat WKP PE-LQ-WD and Gluadin® Kera-PLM with mass ratios of 1:9 and 1:1 at 2 wt% total protein content added to the silicone-free shampoo brought a higher contact angle and a lower frictional coefficient than commercial silicone-free shampoo without proteins. In addition, the compound proteins also have the dual effect of adsorbing the hair surface and penetrating deep into the interior of the hair. The combination of cationic modified hydrolyzed protein (e.g., Croquat WKP PE-LQ-WD) and hydrolyzed protein with low molecular weight (e.g., Gluadin® Kera-P LM) at an appropriate mass ratio exhibited a strong synergistic effect on hair conditioning properties. It could provide a significant reference for developing silicone-free hair products with more benefits.

Transdermal codelivery system of resveratrol nanocrystals and fluorouracil@ HP-β-CD by dissolving microneedles for cutaneous melanoma treatment

Cutaneous melanoma is a serious malignant tumor. In recent years, the morbidity and mortality rate of this disease have been continuously and rapidly increasing. Codelivery of natural product resveratrol (RES) and chemical synthetic drug fluorouracil (Fu) is beneficial to achieve reduced toxicity and increased efficacy effects for melanoma therapeutic. However, the poor solubility and bioavailability of the two drugs resulted in deficient effective and systemic toxicity due to high dose administration. Furthermore, these shortcomings also lead to drug resistance and unsatisfactory patient compliance. In this study, a codelivery dissolvable microneedle (DMN) patch was designed by carrying RES nanocrystals and Fu@hydroxypropyl-beta-cyclodextrin (HP-β-CD) for a synergistic effect. The RES nanocrystals exhibited satisfied solubility and dissolution properties in vitro, and the Fu inclusion compound with HP-β-CD resulted in satisfactory drug loading. Laser confocal fluorescence microscopy demonstrated uniform distribution of both drugs in the DMN. Mechanical tests and in vitro and in vivo puncture tests of the DMN exhibited great mechanical and insertion properties. The DMN in the Franz diffusion cell test released 84.13 ± 5.19 % of the RES and 90.65 ± 18.23 % of the Fu at 12 h. Additionally, for in vivo release, the DMN released 81.24 ± 5.23 % of the RES and 72.48 ± 7.82 % of the Fu. All excipients of the DMN presented high biosafety. In conclusion, codelivery of RES nanocrystals and Fu@HP-β-CD by the DMN presented an excellent pharmaceutical and biological properties and satisfied potential clinical application prospects for cutaneous melanoma treatment.

Rhizoid-associated cyanobacteria are the main contributors to biological nitrogen fixation in restored moss crust

Biological soil crusts (biocrusts) are associations of highly specialized communities and topsoil particles and play important ecosystem functions in drylands. Biotic interactions affected the assemblage of biocrust communities and thus their ecological function and adaption abilities. As the dominant organism in the moss crust, moss plays a significant role in modifying the microenvironment and releasing carbon, and this may affect the distribution and metabolic activity of diazotrophic cyanobacteria, which are the dominant diazotrophs. In this study, we examined the contribution of diazotrophic cyanobacteria to nitrogenase activity (NA) by investigating the NA under different light conditions and estimated the proportion of diazotrophic cyanobacteria from total diazotrophs. We tested the spatial distribution of cyanobacteria using laser confocal fluorescence microscopy (CLSM). Additionally, we separated diazotrophs in moss crust into free-living and moss-associated diazotrophs and compared the NA, nifH gene abundances and their transcripts, and the community composition of free-living and moss-associated diazotrophs. The light-dependent characteristic of NA and dominance of heterocystous cyanobacteria (accounting for over 90% of total nifH sequences at both the DNA and transcript levels) suggested a significant contribution of heterocystous cyanobacteria to NA. Cyanobacteria were mainly distributed in a few millimeters of subsoil in which abundant cyanobacteria were associated with moss rhizoids. The NA rates of moss-associated diazotrophs were comparable to those of moss crust and approximately 4–8 times higher than those of free-living diazotrophs, indicating the major role of moss-associated diazotrophs in nitrogen fixation. The community composition of moss-associated diazotrophs was similar to that of free-living diazotrophs. Moss-associated diazotrophs have a higher NA per copy of the nifH gene and transcript than free-living diazotrophs, proving the synergistic association of moss and cyanobacteria with NA.

The Impact of Al2O3 Particles from Grit-Blasted Ti6Al7Nb (Alloy) Implant Surfaces on Biocompatibility, Aseptic Loosening, and Infection.

For the improvement of surface roughness, titanium joint arthroplasty (TJA) components are grit-blasted with Al2O3 (corundum) particles during manufacturing. There is an acute concern, particularly with uncemented implants, about polymeric, metallic, and corundum debris generation and accumulation in TJA, and its association with osteolysis and implant loosening. The surface morphology, chemistry, phase analysis, and surface chemistry of retrieved and new Al2O3 grit-blasted titanium alloy were determined with scanning electron microscopy (SEM), X-ray energy-dispersive spectroscopy (EDS), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and confocal laser fluorescence microscopy, respectively. Peri-prosthetic soft tissue was studied with histopathology. Blasted retrieved and new stems were exposed to human mesenchymal stromal stem cells (BMSCs) for 7 days to test biocompatibility and cytotoxicity. We found metallic particles in the peri-prosthetic soft tissue. Ti6Al7Nb with the residual Al2O3 particles exhibited a low cytotoxic effect while polished titanium and ceramic disks exhibited no cytotoxic effect. None of the tested materials caused cell death or even a zone of inhibition. Our results indicate a possible biological effect of the blasting debris; however, we found no significant toxicity with these materials. Further studies on the optimal size and properties of the blasting particles are indicated for minimizing their adverse biological effects.

Surfactin molecules with a cone-like structure promote the formation of membrane domains with negative spontaneous curvature and induce membrane invaginations

Surfactin uniquely influences lipid bilayer structure by initially inducing membrane invaginations before solubilization. In this study, we exposed DOPC giant vesicles to various surfactin concentrations at different temperatures and observed surfactin-induced membrane invaginations by using differential interference contrast and confocal laser fluorescence microscopy. These invaginations were stable at room temperature but not at higher temperatures. Surfactin molecules induce membrane nanodomains with negative spontaneous curvature and membrane invaginations despite their intrinsic conical shape and intrinsic positive curvature. Considering the experimentally observed capacity of surfactin to fluidize lipid acyl chains and induce partial dehydration of lipid headgroups, we propose that the resulting surfactin-lipid complexes exhibit a net negative spontaneous curvature. We further conducted 3D numerical Monte Carlo (MC) simulations to investigate the behaviour of vesicles containing negative curvature nanodomains within their membrane at varying temperatures. MC simulations demonstrated strong agreement with experimental results, revealing that invaginations are preferentially formed at low temperatures, while being less pronounced at elevated temperatures. Our findings go beyond the expectations of the Israelachvili molecular shape and packing concepts analysis. These concepts do not take into account the influence of specific interactions between neighboring molecules on the inherent shapes of molecules and their arrangement within curved membrane nanodomains. Our work contributes to a more comprehensive understanding of the complex factors governing vesicle morphology and membrane organization and provides insight into the role of detergent-lipid interactions in modulating vesicle morphology.

Optimizing the accessibility of zeolite Y on FCC catalyst to improve heavy oil conversion capacity

Optimizing the accessibility of zeolite Y, the main active component in fluid catalytic cracking (FCC) catalyst, is a crucial strategy to improve the heavy oil cracking performance of FCC catalysts. Four FCC catalysts with different Y zeolite content were prepared via in-situ crystallization method by adjusting the crystallization time. The distribution characteristics of the Y zeolite in in-situ crystallized catalysts have been characterized by scanning electron microscopy (SEM), laser confocal fluorescence microscopy and high-resolution field emission scanning electron microscopy (HR-FESEM). A unique structural feature of the FCC catalysts has been achieved, with the Y zeolite component mainly distributing in the outer layer of microspheres. The results of adsorption rate of macromolecule probes reveal that the appropriate zeolite layer thickness and topology were beneficial to the mass transfer performance and acid accessibility of FCC catalyst. The effective utilization ratios of zeolite in the novel FCC catalysts for heavy oil cracking on advanced catalyst equipment (ACE) is in good correspondence with the mass transfer performance of macromolecules and the accessibility of acid sites. It could be confirmed that the excellent heavy oil conversion performance of a FCC catalyst does not depend on the content of zeolite, but rather on the distribution characteristics are in favor of the accessibility of zeolite components.

The association of BRAF V600E gene mutation with proliferative activity and histopathological characteristics of congenital melanocytic nevi in children

BackgroundA lot of congenital melanocytic nevi (CMN) carry the somatic mutation in the oncogene BRAF V600E. But the detailed histopathologic characteristics and the proliferative activity of CMN with BRAF V600E gene mutation have not been systematically documented. ObjectiveTo identify the proliferative activity and histopathological features correlating them with BRAF V600E gene mutation status in CMN. MethodsCMN were retrospectively identified from the laboratory reporting system. Mutations were determined by Sanger sequencing. The CMN were divided into a mutant group and control group according to whether there was BRAF gene mutation and were strictly matched according to gender, age, nevus size, and location. Histopathological analysis, analysis of Ki67 expression by immunohistochemistry and laser confocal fluorescence microscopy were performed. ResultsThe differences in Ki67 index, the depth of nevus cell involvement and the number of nevus cell nests between the mutant group and the control group was statistically significant, with p-values of 0.041, 0.002 and 0.007, respectively. Compared with BRAF V600E negative nevi, BRAF V600E positive nevi often exhibited predominantly nested intraepidermal melanocytes, and larger junctional nests, but the difference in this data sets were not statistically significant. The number of nests (p = 0.001) was positively correlated with the proportion of Ki67 positive cells. Study limitationsA small sample of patients were included and there was no follow-up. ConclusionsBRAF V600E gene mutations were associated with high proliferative activity and distinct histopathological features in congenital melanocytic nevi.

Evaluation of the effects of asphalt binder aging degree on the curing, compatibility, and mechanical behaviors of epoxy asphalt binders

The application of epoxy asphalt binder in hot mix plant recycling for reclaimed asphalt pavement (RAP) can significantly increase the utilization rate of waste material on the premise of ensuring the service quality of the asphalt pavement. However, the development of epoxy asphalt binder modified RAP is hindered by the limited understanding of the effect of aging degree and types variability of asphalt binder in RAP on the properties of epoxy asphalt binder. Therefore, short-, long- and ultra-long- term aging tests of base asphalt and SBS modified asphalt binder were conducted through rolling thin film oven (RTFO), pressure aging vessel (20 h and 40 h) tests to obtain the asphalt binders with different aging extent, and then the binders were mixed with two epoxy systems to prepare epoxy asphalt binders, respectively. Next, the effects of asphalt binders aging degree on curing, compatibility, and mechanical behaviors characteristics of epoxy asphalt binders were investigated from chemical, microscopic, mechanical, and rheological perspectives based on Brookfield viscosity, Fourier transform infrared (FTIR), laser confocal fluorescence microscopy (LSCM), Shore hardness, direct tensile, strain sweep, and frequency sweep tests. The viscosity results indicated that a higher asphalt binder aging degree brought about a faster increase in the viscosity of epoxy asphalt binders. The construction allowable time of type-A epoxy asphalt binder of at least 3.5 h was higher than that of type-B. The conversion degree model constructed with the 917 cm−1 peak area as a parameter showed that the epoxy asphalt binder prepared by the asphalt binder with a more severe aging degree presented a higher conversion degree within the same curing time since the reaction space between epoxy and curing agent reactive groups was compressed by aggregated aged asphalt binder. The LSCM and Shore hardness results suggested that the aging asphalt binders induced the phase separation between asphalt and epoxy resin. With the asphalt binders aging, the distribution of asphalt phase in the epoxy system changed from black dots (12–15 um) to island-shaped structures (50–98 um), and eventually transferred a neuron-shaped network structure to connect island-shaped synaptosomes to block the continuity of three-dimensional network structure of epoxy resin. The rheological tests showed that the linear viscoelastic (LEV) region of the epoxy asphalt binders changed with the asphalt binders aging degree, and a shear strain of 0.01% ensured that specimens was within the LEV region. The effects of asphalt binders aging degree on the complex modulus and phase angle master curves of type-A epoxy asphalt binders mainly reflected the low frequency region (10−4 to 10−1 Hz), while the effects on type-B epoxy asphalt binders were in the whole frequency range (10−4 to 103 Hz).

Wattle-Bark-Tannin-Derived Carbon Quantum Dots as Multi-Functional Nanomaterials for Intelligent Detection of Cr6+ Ions, Bio-Imaging, and Fluorescent Ink Applications

Using natural wattle bark tannin, a kind of stable, nontoxic, fluorescent carbon quantum dots (CQDs) was hydrothermally fabricated and applied as a multifunctional fluorescent nanomaterial for the determination of heavy metal ions (Cr6+ and Co2+), bio-imaging, and fluorescent ink applications. The CQDs could be assembled with polyvinyl alcohol (PVA) through hydrogen bonding to yield a composite fluorescent hydrogel that can be integrated with a smartphone to constitute a novel and convenient intelligent detection system for the rapid (10 s) real-time monitoring of Cr6+. Both the CQDs (with 1.37 μM of LOD (limit of detection)) and CQDs-PVA hydrogel (with 3.36 mg/L of LOD) sensing systems produced fast, sensitive, and selective response to Cr6+ based on the internal filtering effect and electron-transfer effect. The practicability of the CQDs-PVA fluorescence sensor was demonstrated by determining tap water and tanning wastewater samples. Based on their satisfactory fluorescence stability and solubility, the CQDs were further used for HeLa-cell imaging and as fluorescent ink for information encryption. When HeLa cells were incubated for 24 h in CQDs at a high concentration of 200 mg/L, the cell survival remained as high as 90%, and a clear fluorescence image was observed under a laser confocal fluorescence microscope. The CQD solution could be written directly as fluorescent ink on TLC (thin layer chromatography) paper, and the handwritings were invisible after drying, confirming their application potential in the field of information encryption. In summary, the present CQDs derived from wattle bark tannin exhibited excellent stability, sensitivity, and specificity in heavy metal ion sensing and also had great potential in bio-imaging and information encryption applications.

Effects of potassium channel knockdown on peripheral blood T lymphocytes and NFAT signaling pathway in Xinjiang Kazak patients with hypertension

ABSTRACT Backgroud and aim T lymphocytes are involved in the occurrence and development of essential hypertension, and potassium channels are thought to be critical for lymphocyte activation. This study is to examine the roles of the voltage-gated potassium channels (Kv1.3) and calcium-activated potassium channels (KCa3.1) in peripheral blood T lymphocytes in Kazakh hypertensive patients of Xinjiang, China, mainly focusing on the effects of these channels on nuclear factor of activated T cells (NFAT) and inflammatory cytokines of T lymphocytes. Method Kv1.3 and KCa3.1 gene silencing were performed in cultured T lymphocytes from Kazakh patients with severe hypertension. T cell proliferation after gene silencing was measured using CCK-8. The mRNA and protein expression levels were measured using RT-qPCR and Western blot analysis, respectively. Nuclear translocation of NFAT was observed using laser confocal fluorescence microscopy. Inflammatory cytokine levels were detected with ELISA. Results Compared with control group, gene silencing of Kv1.3 and KCa3.1 respectively inhibited the proliferation of T cells. Moreover, compared with the control group, the mRNA expression levels of NFAT, IL-6 and IFN-γ were significantly decreased after gene silencing. Furthermore, the NFAT protein expression level was significantly down-regulated. In addition, the levels of IFN-γ and IL-6 in the cell culture supernatant were significantly decreased. Conclusion Both Kv1.3 and KCa3.1 potassium channels activated T lymphocytes and enhanced the cytokine secretion possibly through CaN/NFAT signaling pathway, which may in turn induce micro-inflammatory responses and trigger the occurrence and progression of hypertension.

Influence of bioceramic intracanal medication on the bond strength of bioceramic root canal sealer.

To investigate the influence of the remaining volume of a new intracanal medication based on bioceramic compounds on the bond strength (BS) and formation of an adhesive interface between calcium silicate-based and epoxy resin-based root canal sealers. For this purpose, the specimens were distributed according to the intracanal medication (n = 26): Bio-C Temp (BCT) and Ultracal XS (UXS). The roots were scanned in microCT, and after 7 days, the medication was removed. Then a new scan was performed to evaluate the volume of medication remaining. Subsequently, 40 specimens were redistributed into 2 subgroups (n = 10) and filled according to the sealer used: AH Plus (AHP) and Bio-C Sealer (BCS), to assess the bond strength by using the push-out test, and the adhesive interface by confocal laser fluorescence microscopy (CLSM) and scanning electron microscopy (SEM). The t test showed a smaller remainder of BCT (1.77 ± 0.86) compared with UXS (10.47 ± 5.78), irrespective of the root third evaluated. The BS showed that teeth with BCT + BCS had higher bond strength values (3.70 ± 1.22) when compared to the other groups: BCT + AHP (2.15 ± 1.07), UXS + BCS (3.18 ± 1.09) and UXS + AHP (2.11 ± 1.02) (p<0.001). The cervical third had higher BS when compared with the middle and apical thirds (p < 0.001), and higher number of adhesive failures. The adhesive interface in SEM and CLSM images showed better adaptation for the association between BCT + BCS. Intracanal medication and silicate-based endodontic sealer appeared to interact chemically by forming a biomineralizing layer, allowing for an increase in the bond strength and forming an adhesive interface between the materials, with no or less gap formation.

Fabrication of hydroxyapatite porous body with connective pores using the self-networking property of rice starch powder by heat treatment

• Porous hydroxyapatite ceramics was fabricated. • Gelatinization-retrogradation phenomena of starch was used as pore-networking agent. • Randomly-developed open pores were introduced into the hydroxyapatite matrix. • Pore structure was characterized by confocal scanning laser fluorescence microscopy. Hydroxyapatite porous ceramics were fabricated utilizing the property called gelatinization-retrogradation in which starch particles used as a pore-forming agent form a self-network structure by heat treatment. The pore structures in the hydroxyapatite samples were observed by combining the immersion transmission method and a confocal scanning laser fluorescence microscope. The three-dimensional image created by stacking the cross-sectional observation images showed that randomly-developed open pores were introduced into the matrix.

Prodigiosin from Serratia Marcescens in Cockroach Inhibits the Proliferation of Hepatocellular Carcinoma Cells through Endoplasmic Reticulum Stress-Induced Apoptosis.

Hepatocellular carcinoma (HCC) is the most common primary liver malignant tumor, and the targeted therapy for HCC is very limited. Our previous study demonstrated that prodigiosin(PG), a secondary metabolite from Serratia marcescens found in the intestinal flora of cockroaches, inhibits the proliferation of HCC and increases the expression of CHOP, a marker protein for endoplasmic reticulum stress (ERS)-mediated apoptosis, in a dose-dependent manner. However, the mechanisms underlying the activity of PG in vivo and in vitro are unclear. This study explored the molecular mechanisms of PG-induced ERS against liver cancer in vitro and in vivo. The apoptosis of hepatocellular carcinoma cells induced by PG through endoplasmic reticulum stress was observed by flow cytometry, colony formation assay, cell viability assay, immunoblot analysis, and TUNEL assay. The localization of PG in cells was observed using laser confocal fluorescence microscopy. Flow cytometry was used to detect the intracellular Ca2+ concentration after PG treatment. We found that PG could promote apoptosis and inhibit the proliferation of HCC. It was localized in the endoplasmic reticulum of HepG2 cells, where it induces the release of Ca2+. PG also upregulated the expression of key unfolded response proteins, including PERK, IRE1α, Bip, and CHOP, and related apoptotic proteins, including caspase3, caspase9, and Bax, but down-regulated the expression of anti-apoptotic protein Bcl-2 in liver cancer. Alleviating ERS reversed the above phenomenon. PG had no obvious negative effects on the functioning of the liver, kidney, and other main organs in nude mice, but the growth of liver cancer cells was inhibited by inducing ERS in vivo. The findings of this study showed that PG promotes apoptosis of HCC by inducing ERS.

Analysis of the influencing mechanism of the freeze–thawing cycles on in vitro chicken meat digestion based on protein structural changes

The effect of freeze–thaw (F–T) cycles on the digestive and protein structural properties of chicken breasts during in vitro digestion was investigated. With F–T cycles increased, the hardiness, chewiness, and shear force of chicken breasts increased, whereas the digestibility decreased, and particle size of digestive samples increased was evidenced by laser particle size analyzer and confocal fluorescence microscopy. The digestibility of the fifth F–T cycle samples in pepsin and pepsin/trypsin decreased by 25.99% and 11.82% compared to fresh samples, respectively. During F–T cycles, the disruption of protein structure was confirmed by the α-Helix decrease, β-sheet increase, the intrinsic tryptophan intensities decrease and a redshift in the maximum value. Therefore, F–T cycles destroyed protein structure and induced the protein aggregation, resulting in chicken that was difficult to digest in the simulated digestion. The phenomenon became more acute as the number of F–T cycles increased.

Toward quantitative experiment using hydrothermal diamond anvil cell: Solubility of sylvite up to 1.6 GPa

Potassium chloride is a common component in geofluids and sylvite is a broadly observed daughter mineral in fluid inclusions along with halite, especially in skarn- and porphyry-type deposits. Interpretation of fluid inclusion microthermometric results requires knowledge of high pressure and temperature (P-T) solubility of salts, which is, however, incomprehensive for sylvite in the KCl-H2O system. In this study, the high P-T solubility of sylvite is determined within the compositional (salinity) range of 38–62 wt% KCl, using a new method that applies the three-dimensional reconstruction function of confocal laser fluorescence microscopy (CLFM) to hydrothermal diamond anvil cell (HDAC) experiments. This method is used to retrieve the volumes of phases (i.e., sylvite, aqueous solution and bubble) in the sample chamber of HDAC and thus to achieve quantitative analysis of sylvite solubility at elevated P-T. Combining experimental results in this study with those published in previous literature, the P-T dependence of the solubility of sylvite is retrieved and applied to data interpretation of microthermometric results. For inclusions containing both halite and sylvite daughter minerals, the commonly used approach for evaluating fluid salinity and P is to simplify the inclusions as the binary NaCl-H2O system. This simplification would lead to underestimation of fluid salinities by up to ~34% and homogenization P by up to 0.25 GPa, for fluid inclusions having relatively high T of daughter mineral dissolution, which are typical of magmatic-hydrothermal ore deposits.

A highly selective and sensitive “turn-on” fluorescent probe for Fe2+ and its applications

The determination of Fe2+ in living systems is of great significance owing to its crucial roles in the biological systems. Herein, a turn-on fluorescent probe based on N-oxide for detecting Fe2+ is designed and synthesized. The probe shows excellent sensitivity to Fe2+ with the detection limit of 0.21 μM. The coexistence interfering species have little influence on the fluorescence signal of the probe for detection of Fe2+. This turn-on response is accomplished via a redox-involved reaction triggered by Fe2+. The probe showed excellent cell membrane permeability and has been successfully used to monitor Fe2+ in living cell and zebrafish with a laser confocal fluorescence microscope and detect Fe2+ in real water samples.

Transcytosis mechanisms of cell-penetrating peptides: Cation-independent CC12 and cationic penetratin.

Cell-penetrating peptides (CPPs) can aid in intracellular and in vivo drug delivery. However, the mechanisms of CPP-mediated penetration remain unclear, limiting the development and further application of CPPs. Flow cytometry and laser confocal fluorescence microscopy were performed to detect the effects of different endocytosis inhibitors on the internalization of CC12 and penetratin in ARPE-19 cells. The co-localization of CPPs with the lysosome and macropinosome was detected via an endocytosis tracing experiment. The flow cytometry results showed that chlorpromazine, wortmannin, cytochalasin D, and the ATP inhibitor oligomycin had dose-dependent endocytosis-inhibitory effects on CC12. The laser confocal fluorescence results showed that oligomycin had the most significant inhibitory effect on CC12 uptake; CC12 was co-located with the lysosome, but not with the macropinosome. For penetratin, cytochalasin D and oligomycin had obvious inhibitory effects. The laser confocal fluorescence results indicated that oligomycin had the most significant inhibitory effect on penetratin uptake; the co-localization of penetratin with the lysosome was higher than that with the macropinosome. Cation-independent CC12 and cationic penetratin may be internalized into cells primarily through caveolae and clathrin-mediated endocytosis, and they are typically dependent on ATP. The transport of penetratin could be partly achieved through the direct transmembrane pathway, as the positive charge of penetratin interacts with the negative charge of the cell membrane, and partly through the endocytic pathway.

Photodynamic evaluation of triazine appended porphyrins as anti-leishmanial and anti-tumor agents

Many recent advancements in porphyrin based photodynamic therapy are being treated as revolutionary in the field of medicinal chemistry, as they offer effective and viable solutions for many serious diseases including cancer. Motivated by the exceptional bioactivity of triazine moieties, we designed 4,6-diamino-1,3,5-triazine appended 5,10,15,20-tetrakis(4-hydroxyphenyl)porphyrin (THP) and it's metal derivatives; adopting a mild and effective microwave assisted protocol. All the synthesized derivatives were evaluated in-vitro for their anti-proliferative activity against human malignant melanoma (A375), human breast cancer (MCF-7) and promastigote form of Leishmania donovaniby MTT assay in the presence of light. The study revealed better inhibitory activity of the derivatives under illumination, compared to the parent compound. The compounds were found to be non-toxic to normal cells at their respective concentrations of anti-proliferative action, in both dark and light. The cellular uptake of the compounds in tumor cell was confirmed using laser confocal fluorescence microscopy. In order to establish the mechanism of action, the singlet oxygen generation efficiency was also investigated. The superior inhibitory activity of the zinc derivative has been indicated by its highest singlet oxygen generation efficiency (ΦΔ = 0.69).

Fabrication of protein-coated titanium dioxide nanoparticles for cellular uptake fluorescence imaging and treatment of colorectal cancer

Titanium dioxide (TiO2) coated with different proteins has exhibited exceptional bio-compatibility, leading to various biomedical engineering applications. With the use of green and chemical reduction methods, we fabricated Titanium dioxide nanoparticles that were protein-coated. Bovine serum albumin (BSA), lysozyme proteins, zein, and collagen have been used to coat titanium dioxide-aryl nanoparticles of the form TiO2-NPs. However, in both cases, no catalysts or other stabilizing agents were used. These images of TiO2-NPs fabricated using the green method show high crystallinity. It is a malignant colorectal tumour with dysfunctional cellular processes that cause colorectal cancer cells. It is hoped that studies employing SW1417 cells would give mechanistic ideas on the specifics of the amplification in cancers. This was done by flow cytometry utilizing and laser confocal fluorescence microscopy (LCFM) on the SW1417 colorectal cell line. Of the protein-coated Titanium dioxide nanoparticles fabricated green methods, BSA@TiO2-NPs were the most readily absorbed. Of all TiO2-NPs, lysozyme@TiO2-NPs fabricated by the chemical reduction technique were the most effectively internalized by SW1417 cells out of TiO2-NPs types. However, TiO2-NPs fabricated by the green methodology were coated with zein and lysozyme and tiny. A hydrophobic covering is also on the two nanoparticles. There is a possibility that the variation in hydrophobicity and charge affected the internalization process. Colorectal diagnostic and therapeutic compounds might be synthesized from those coated nanoparticles that were effectively internalized.

Hydroclimate response in Texas and Gulf of Mexico to rapid warming during the last deglacial: High-resolution speleothem proxy and monitoring evidence

Cave calcite (i.e., speleothem) proxy records that span abrupt warming events, such as those of the last deglacial, may prologue regional responses of hydroclimate to 21st century warming. Proxy reliability is advanced by an understanding of the modern hydrologic system and controls on calcite growth. Here we integrate monitoring of Cave CWN in central Texas with high-resolution imaging and δ18O analysis of Stalagmite CWN-4 to understand how regional hydroclimate changed during a rapid deglacial warming event - the transition from Heinrich Stadial 1 (HS1) to the Bølling-Allerød (BA) warm period. This event is recorded in CWN-4 as a rapid negative δ18O excursion of 1.7‰ over ∼210 years (HS1-BA δ18O excursion = ‘HBO’), followed by a 6-fold growth rate increase.Monitoring demonstrates that modern calcite grows during cool seasons, growth rates follow drip rate, and drip water and calcite δ18O reflect the δ18O of recharge. Because drip waters have interannual epikarst residence times, their δ18O values are well-mixed averages of their Gulf of Mexico (GoM) moisture source δ18O values, which rapidly decreased during Laurentide Ice Sheet (LIS) melting events. Confocal laser fluorescence microscopy imaging indicates that growth lamina are seasonal within U-series age constraints. The timing of the HBO (14.75 ± 0.08 to 14.54 ± 0.08 ka), corresponds with 1) the onset of Bølling warming in Greenland (14.64 ka), 2) AMOC reinvigoration (∼14.61–14.25 ka), 3) Meltwater Pulse 1a (14.65–14.31 ka), and 4) Meltwater Flood 3 (MWF-3) - the most intense episode of LIS meltwater discharge into the GoM (14.97–14.46 ka). The rapid increase in growth rates following the HBO indicates an abrupt transition to a wetter hydroclimate in central Texas following these regional and global events.The HBO δ18O excursion occurred monotonically, consistent with meltwater discharge being continuous. This excursion also corresponds with climate change recognized across the HS1-BA transition in marine and terrestrial records influenced by changes in North Atlantic climate, AMOC, and ITCZ migrations. We infer that central Texas deglacial hydroclimate largely followed changes in AMOC, linked via meltwater input to the GoM. Periods of fast (slow) speleothem growth coincide with warmer (cooler) North Atlantic episodes, with the exception of the HS1 to BA transition. For this transition, slow growth rates during MWF-3 may reflect protracted cooling of GoM SST and corresponding atmospheric moisture reduction. The common timing and duration of the HBO with shifts toward wetter conditions across the HS1-BA transition in many northern Hemisphere climate records supports common ocean-atmosphere teleconnections related to AMOC intensity.