Protoporphyrin IX-mediated sonodynamic action induces apoptosis of K562 cells

Objectives

The present study aims to investigate apoptosis of human leukemia K562 cells induced by protoporphyrin IX (PpIX)-mediated sonodynamic therapy (PpIX-SDT).

Methods

The uptakes of intracellular PpIX in K562 cells were detected by flow cytometry. The sub-cellular localization of PpIX was imaged by confocal microscope. The cytotoxic effect of PpIX-SDT was assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenylter-trazolium bromide tetrazolium) assay. Apoptosis was evaluated by chromatin condensation with DAPI (4′-6-diamidino-2-phenylindole) staining, decrease of mitochondria membrane potential (MMP), re-distribution of Bax, and the expression changes of the key apoptosis-associated protein (Caspase-3 and polypeptide poly (ADP-robose) polymerase). The possible mechanism of SDT-induced apoptosis was investigated by detecting by intracellular ROS (reactive oxygen species) generation and effect of ROS scavenger-NAC (N-acetylcysteine) on SDT induced apoptosis.

Results

The intracellular PpIX increased quickly within 2 h after PpIX administration and PpIX mainly localized in the mitochondria. Compared with PpIX alone and ultrasound alone groups, the synergistic cytotoxicity of PpIX plus ultrasound was significantly boosted. In addition, the ultrasound induced some extent of chromatin condensation and MMP loss was greatly enhanced by the presence of 2 μg/ml PpIX, where PpIX alone treatment showed no or only slight effect. Time-dependent Bax translocation, caspase-3 activation and PARP cleavage were detected in SDT treatment groups. Besides, intracellular ROS production was significantly enhanced after SDT, and the general ROS scavenger NAC could obviously alleviate the SDT-caused cell viability loss, MMP loss, Bax redistribution and nuclear changes.

Conclusions

These results indicated that PpIX-mediated sonodynamic action could induce apoptosis on K562 cells, and the intracellular ROS was involved in the PpIX-SDT induced apoptosis.     

Hypocrellin B-mediated sonodynamic action induces apoptosis of hepatocellular carcinoma cells

Objective

The present study aims to investigate apoptosis of hepatocellular carcinoma cells induced by hypocrellin B-mediated sonodynamic action.

Methods

The hypocrellin B concentration was kept constant at 2.5 μM and cells from the hepatocellular carcinoma HepG2 cell line were exposed to ultrasound with an intensity of 0.46 W/cm² for 8 s. Cell cytotoxicity was quantified using an MTT assay 24 h after sonodynamic therapy (SDT) of hypocrellin B. Apoptosis was investigated using a flow cytometry with Annexin V-FITC and propidium iodine staining. Intracellular reactive oxygen species (ROS) levels were detected using a flow cytometry with 2,7-dichlorodihydrofluorecein diacetate (DCFH-DA) staining.

Results

The cytotoxicity of hypocrellin B-mediated sonodynamic action on HepG2 cells was significantly higher than those of other treatments including ultrasound alone, hypocrellin B alone and sham treatment. Flow cytometry showed that hypocrellin B-induced sonodynamic action markedly enhanced the apoptotic rate of HepG2 cells. Increased ROS was observed in HepG2 cells after being treated with hypocrellin B-mediated sonodynamic action.

Conclusions

Our data demonstrated that hypocrellin B-mediated sonodynamic action remarkably induced apoptosis of HepG2 cells, suggesting that apoptosis is an important mechanism of cell death induced by hypocrellin B-mediated SDT.     

Sonodynamically induced anti-tumor effect with protoporphyrin IX on hepatoma-22 solid tumor

Objective

The purpose of this study was to evaluate sonodynamically induced anti-tumor effect of protoporphyrin IX (PPIX) in mice bearing hepatoma-22 (H-22) solid tumors, and the possible in vivo cell damage mechanism was also investigated.

Methods

The pharmacokinetics of PPIX was analyzed in plasma, skin, muscle and tumor of H-22 bearing mice. Tumors were irradiated with ultrasound (1.43 MHz, I_SATA 3 W/cm², 3 min) for three times at 8, 12 and 24 h after 5.0 mg/kg PPIX administration, respectively. The anti-tumor effects of sonodynamic therapy (SDT) were estimated by the tumor inhibition ratio (volume and weight). The bio-effects of SDT were evaluated by hematoxylin and eosin (H&E) staining, transmission electron microscope (TEM) observation, lipid peroxidation (LPO) measurement and anti-oxidative enzymes (glutathione peroxidase (GSH-PX), catalase (CAT) and superoxide dismutase (SOD)) assay.

Results

A significant anti-tumor effect was obtained by PPIX-mediated sonodynamic therapy (PPIX-SDT). At the fifteenth day after PPIX-SDT, the tumor growth and tumor weight inhibition ratios were 53.84% and 45.86%, respectively. In addition, the structure of tumor tissues and the anti-oxidative enzymes were obviously destroyed after SDT treatment.

Conclusions

A biochemical mechanism was involved in PPIX-SDT in vivo, and the free radicals produced by the synergistic treatment destroying the anti-oxidative system of tumor cells in vivo may play an important role in this action. Also, the thermal effect could not be excluded in inducing damage of cellular structures, like membrane disruption and chromatin condensation under current evaluation in this paper.     

Apoptosis of ovarian cancer cells induced by methylene blue-mediated sonodynamic action

Objective

The present study aims to investigate apoptosis of ovarian cancer cells induced by methylene blue (MB)-mediated sonodynamic therapy (SDT).

Methods

The MB concentration was kept constant at 100 μM and ovarian cancer HO-8910 cells were exposed to ultrasound therapy for 5 s with an intensity of 0.46 W/cm². The cytotoxicity was investigated 24 h after MB-mediated sonodynamic action. Apoptosis was analyzed using a flow cytometer with Annexin V-FITC and propidium iodine (PI) staining as well as fluorescence microscopy with Hoechst 33258 staining. Intracellular reactive oxygen species (ROS) level was measured by flow cytometer with 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) staining.

Results

The cytotoxicity of MB-mediated SDT on HO-8910 cells after MB-mediated SDT was significantly higher than those of other treatments including ultrasound alone, MB alone and sham treatment. Flow cytometric analysis showed a significant increase in the early and late apoptotic cell populations by MB-mediated SDT of HO-8910 cells. Nuclear condensation and increased ROS levels were also found in HO-8910 cells treated by MB-mediated SDT.

Conclusions

Our findings demonstrated that MB-mediated sonodynamic action significantly induced apoptosis of HO-8910 cells and an increase in intracellular ROS level. This indicates that apoptosis is an important mechanism of cell death induced by MB-mediated SDT. Thus, MB-mediated SDT might be a potential therapeutic strategy for combating ovarian cancer.     

Augmentation of the Porcine Vocal Fold Using Autologous Composite Cervical Fascia and Fat Graft. Comparison Between the Transmuscular and Submuscular Approaches

Objectives

To assess the integration of an autologous composite fascia and fat graft implanted into the lamina propria of a porcine vocal fold using two different approaches.

Study Design

An experimental prospective study on the porcine larynx was conducted at a tertiary research institution.

Methods

An external cervical approach was used to expose the thyroid cartilage of 24 healthy minipigs under general anesthesia. The composite fascia/fat graft was implanted through two distinct approaches, transmuscular and submuscular. Animals were sacrificed at 7, 30, 90, and 180 days for macroscopic and histological study of the larynx. Graft integration and local inflammatory response were studied.

Results

The survival rate of the experimental model was 100% and all animals had local inflammatory response to the surgical procedure. Only 41.7% of the grafts placed inside the thyroarytenoid (TA) muscle fibers were identified postmortem, whereas 83.3% of the submuscular grafts remained intact.

Conclusions

Graft incorporation was better and there was less inflammation when the architecture of the TA muscle fibers was preserved. Graft extrusion was observed in the most of the cases where it was placed inside the muscle.     

Photodynamic Therapy for Cancer Cells Using a Flash Wave Light Xenon Lamp

We determined photodynamic therapy (PDT) efficacy using a flash wave (FW) and a continuous wave (CW) light, of which the fluence rate was 70 W/cm², for murine thymic lymphoma cells (EL-4) cultivated in vitro. The irradiation frequency and the pulse width of the FW light were in the range of 1–32 Hz and less than one millisecond, respectively. 5-Aminolevulinic acid-induced protoporphyrin IX (ALA-PpIX) was used as a photosensitizer. When EL-4 with ALA administration was irradiated by the light for 4 h (irradiation fluence: 1.0J/cm²), the survival rate of EL-4 by the FW light was lower than that by the CW light, except for the FW light with irradiation frequency of 32 Hz, and decreased gradually with decreasing irradiation frequency. Moreover, the FW light, especially at lower irradiation frequency, was superior to the CW light for the generation of singlet oxygen in an aqueous PpIX solution. Therefore, thehigher PDT efficacy for EL-4 of the FW light would be caused by the greater generation of singlet oxygen in the cells.     

The Corolla Polynomial for Spontaneously Broken Gauge Theories

In Kreimer and Yeats (Electr. J. Comb. 41–41, 2013), Kreimer et al. (Annals Phys. 336, 180–222, 2013) and Sars (2015) the Corolla Polynomial \( \mathcal C ({\Gamma }) \in \mathbb C [a_{h_{1}}, \ldots , a_{h_{\left \vert {\Gamma }^{[1/2]} \right \vert }}]\) was introduced as a graph polynomial in half-edge variables \(\{a_{h}\}_{h \in {\Gamma }^{[1/2]}}\) over a 3-regular scalar quantum field theory (QFT) Feynman graph Γ. It allows for a covariant quantization of pure Yang-Mills theory without the need for introducing ghost fields, clarifies the relation between quantum gauge theory and scalar QFT with cubic interaction and translates back the problem of renormalizing quantum gauge theory to the problem of renormalizing scalar QFT with cubic interaction (which is super renormalizable in 4 dimensions of spacetime). Furthermore, it is, as we believe, useful for computer calculations. In Prinz (2015) on which this paper is based the formulation of Kreimer and Yeats (Electr. J. Comb. 41–41, 2013), Kreimer et al. (Annals Phys. 336, 180–222, 2013) and Sars (2015) gets slightly altered in a fashion specialized in the case of the Feynman gauge. It is then formulated as a graph polynomial \(\mathcal C ({\Gamma } ) \in \mathbb C [a_{h_{1 \pm }}, \ldots , a_{h_{\left \vert {\Gamma }^{[1/2]} \right \vert } \vphantom {h}_{\pm }}, b_{h_{1}}, \ldots , b_{h_{\left \vert {\Gamma }^{[1/2]} \right \vert }}] \) in three different types of half-edge variables \( \{a_{h_{+}} , a_{h_{-}} , b_{h}\}_{h \in {\Gamma }^{[1/2]}} \). This formulation is also suitable for the generalization to the case of spontaneously broken gauge theories (in particular all bosons from the Standard Model), as was first worked out in Prinz (2015) and gets reviewed here.     

Mathematical modelling and numerical simulation of the morphological development of neurons

Background

The morphological development of neurons is a very complex process involving both genetic and environmental components. Mathematical modelling and numerical simulation are valuable tools in helping us unravel particular aspects of how individual neurons grow their characteristic morphologies and eventually form appropriate networks with each other.

Methods

A variety of mathematical models that consider (1) neurite initiation (2) neurite elongation (3) axon pathfinding, and (4) neurite branching and dendritic shape formation are reviewed. The different mathematical techniques employed are also described.

Results

Some comparison of modelling results with experimental data is made. A critique of different modelling techniques is given, leading to a proposal for a unified modelling environment for models of neuronal development.

Conclusion

A unified mathematical and numerical simulation framework should lead to an expansion of work on models of neuronal development, as has occurred with compartmental models of neuronal electrical activity.

     

Pitch Characteristics of Homosexual Males

Objective

To investigate the common stereotype that homosexual males show pitch patterns that mirror those of heterosexual females.

Study Design

Static group comparison.

Method

Comparison of speaking fundamental frequency and pitch variation of 30 homosexual males, 56 heterosexual age-matched males, and 54 age-matched heterosexual females as demonstrated in a sample of read speech.

Results

In the homosexual males, average fundamental frequency and pitch variation were significantly higher than in the heterosexual males but also significantly lower than in the heterosexual females.

Conclusions

Results do not confirm the stereotype that gay male speech mirrors the patterns of women’s speech with respect to pitch characteristics. It would seem that the pitch patterns of gay male speakers constitute an example of sociophonetic variation.     

4<Emphasis Type="Italic">p</Emphasis><Superscript>5</Superscript>4<Emphasis Type="Italic">d</Emphasis><Superscript>2</Superscript> configuration in the spectra of rubidium-like Ru VIII and Rh IX ions

The spectra of ruthenium and rhodium excited in a low-inductive vacuum spark have been obtained using a vacuum grazing-incidence spectrograph with a grating of 3600 lines/mm 3 m in radius in the range of 180–350 Å. The bidium-like spectra of Ru VIII and Rh IX were analyzed. Ten spectral lines of the 4p ⁶4d-4p ⁵4d ² transitions are identified and six energy levels in the 4p ⁵4d ² configuration with an excited internal electron were found for each ion for the first time.     

Pharmacokinetics of gadoversetamide injection, a gadolinium-based contrast agent, in pediatric patients

Objective

The pharmacokinetics of gadoversetamide were examined in pediatric patients scheduled to undergo contrast-enhanced MRI of the central nervous system.

Materials and Methods

One hundred patients received an intravenous injection of gadoversetamide at a dose of 0.1 mmol/kg for a contrast-enhanced MRI procedure. A subpopulation of 30 patients were enrolled to evaluate the pharmacokinetics of gadoversetamide in patients 2–11 and 12–18 years of age. Serial blood and urine samples were collected before and after the administration of gadoversetamide.

Results

The terminal half-life, initial concentration and area under the curve assessments for gadoversetamide showed no significant (P>.05) differences between the age groups or the sexes. Although no sex-related differences occurred in the volume of distribution or clearance, significant (P<.05) age-related differences were found, but once corrected for body mass or surface area the differences were no longer significant.

Conclusions

The pharmacokinetic behavior of gadoversetamide was not significantly altered by differences in age or sex in pediatric patients from 2 to 18 years of age. Although significant differences in volumes of distribution, and clearance occurred between the age groups, these differences appeared to depend on body size rather than on age in pediatric patients between 2 and 18 years of age.     

Calculation of Spectra of Neutrons and Charged Particles Produced in a Target of a Neutron Generator

An algorithm for calculating the spectra of neutrons and associated charged particles produced in the target of a neutron generator is detailed. The products of four nuclear reactions ³H(d,n)⁴He, ²H(d,n)³He, ²H(d,p)³H, and ³He(d,p)⁴He are analyzed. The results of calculations are presented in the form of neutron spectra for several emission angles and spectra of associated charged particles emitted at an angle of 180° for a deuteron initial energy of 0.13 MeV.     

Effect of visual target blurring on accommodation under distance viewing

Purpose

To examine the effect of visual target blurring on accommodation.

Methods

We evaluated the objective refraction values when the visual target (asterisk; 8°) was changed from the state without Gaussian blur (15 s) to the state with Gaussian blur adapted [0(without blur)?→?10, 0?→?50, 0?→?100: 15 s each].

Results

In Gaussian blur 10, when blurring of the target occurred, refraction value did not change significantly. In Gaussian blur 50 and 100, when blurring of the target occurred, the refraction value became significantly myopic.

Conclusion

Blurring of the distant visual target results in intervention of accommodation.

     

The effects of Ce6-mediated sono-photodynamic therapy on cell migration,apoptosis and autophagy in mouse mammary 4T1 cell line

Purpose

Sono-Photodynamic therapy (SPDT) is an alternative therapy which claims to enhance the anti-cancer effects by combining sonodynamic therapy (SDT) with photodynamic therapy (PDT). In the present study, we investigated the effects of chlorin e6 (Ce6) mediated SPDT on migration, apoptosis and autophagy in mouse mammary 4T1 cancer cells, and its underlying mechanisms.

Materials

Cell migration was determined by wound healing assay. Apoptosis was analyzed using annexin V-PE/7-ADD staining as well as Hoechst 33342 staining. Changes of mitochondria membrane potential (MMP) was evaluated by flow cytometry. Formation of acidic vesicular organelles (AVOs) during autophagy was observed with fluorescence microscope by MDC staining. Immunofluorescence assays were performed to detect the co-localization of LC3 and Lamp2. Western blotting was employed to analyze the activity of the apoptosis related proteins Caspase-3, PARP, Bax and Bcl-2, as well as the autophagy associated processing of LC3-I to LC3-II and Beclin-1 expression.

Results

Ce6 mediated SPDT further enhanced cell migration inhibition, significantly triggered cell apoptosis, nuclear condensation and MMP drop. Cleaved Caspase-3 and PARP increased dramatically after Ce6-SPDT, accompanied by decreased Bcl-2 expression, while the expression of Bax remained stable. Additionally, AVOs formation, co-localization of LC3 and Lamp2 occurred following Ce6-SPDT and simultaneously accompanied by LC3-II processing and increased Beclin-1 expression.

Conclusions

Ce6-SPDT could enhance cell migration inhibition, and induce mitochondria-dependent apoptosis as well as autophagy in 4T1 cells.     

Conventional and microwave-assisted synthesis of new indole-tethered benzimidazole-based 1,2,3-triazoles and evaluation of their antimycobacterial,antioxidant and antimicrobial activities

A new series of triheterocycles containing indole–benzimidazole-based 1,2,3-triazole hybrids have been synthesized in good yields via a microwave-assisted click reaction. All the compounds were characterized by IR, \(^{1}\hbox {H}\) NMR, \(^{13}\hbox {C}\) NMR and mass spectroscopy and were evaluated for their in vitro antitubercular activity against the Mycobacterium tuberculosis H37Rv strain. Compounds 4b, 4h and 4i displayed highly potent antitubercular activity with MIC 3.125–6.25 \(\upmu \hbox {g}/\hbox {mL}\). The antioxidant potential was evaluated using 2,2-diphenyl-1-picryl hydrazine and \(\hbox {H}_{2}\hbox {O}_{2}\) radical scavenging activity, and compounds 4e,4f and 4g showed excellent radical scavenging activity with \(\hbox {IC}_{50}\) values in the range of 08.50–10.05 \(\upmu \hbox {g}/\hbox {mL}\). Furthermore, the compounds were evaluated for antimicrobial activity against numerous bacterial and fungal strains, and compounds 4b, 4c and 4h were found to be the most promising potential antimicrobial molecules with MIC 3.125–6.25 \(\upmu \hbox {g}/\hbox {mL}\).     

Ni(II) and Zn(II) Complexes Containing Alkynyl Functionalized Salicylaldimine Ligand and Heterocyclic Coligand: Synthesis,Characterization and Luminescence Properties

Some nickel(II) and zinc(II) complexes of the type [Ni(L)(phen/bipy)]X (1a–6a) and [Zn(L) (phen/bipy)]X (1b–6b) (where L = 2-{(E)-[(4-trimethylsilylethynylphenyl)imino]methyl}-4-(4-nitro phenylethynyl)phenol; phen = 1, 10-phenanthroline, bipy = 2, 2´-bipyridine; X = ClO₄ ^?, BF₄ ^?, PF₆ ^?) have been prepared and characterized on the basis of elemental analyses, FTIR, ¹H NMR and mass spectral studies. The molecular structure of L was determined by single crystal X-ray diffraction studies. The electrochemical behaviour of the Ni(II) complexes indicate that the phen complexes appears at more positive potential as compared to those for bipy complexes, as a consequence of its strong π-acidic character. TGA was carried out to study the thermal behavior of the complexes. Room temperature luminescence is observed for all complexes corresponds to π → π* ILCT transition. The size of the counter anion and heterocyclic coligands phen and bipy shows marked effect on emission properties of the complexes.