Photodynamic therapy induces selective extravasation of macromolecules: Insights using intravital microscopy

Photodynamic therapy (PDT) with Visudyne^® acts by direct cellular phototoxicity and/or by an indirect vascular-mediated effect. Here, we demonstrate that the vessel integrity interruption by PDT can promote the extravasation of a macromolecular agent in normal tissue. To obtain extravasation in normal tissue PDT conditions were one order of magnitude more intensive than the ones in tissue containing neovessels reported in the literature.Fluorescein isothiocyanate dextran (FITC-D, 2000 kDa), a macromolecular agent, was intravenously injected 10 min before (LK0 group, n = 14) or 2 h (LK2 group, n = 16) after Visudyne^®-mediated PDT in nude mice bearing a dorsal skin fold chamber. Control animals had no PDT (CTRL group, n = 8). The extravasation of FITC-D from blood vessels in striated muscle tissue was observed in both groups in real-time for up to 2500 s after injection. We also monitored PDT-induced leukocyte rolling in vivo and assessed, by histology, the corresponding inflammatory reaction score in the dorsal skin fold chambers.In all animals, at the applied PDT conditions, FITC-D extravasation was significantly enhanced in the PDT-treated areas as compared to the surrounding non-treated areas (p < 0.0001). There was no FITC-D leakage in the control animals. Animals from the LK0 group had significantly less FITC-D extravasation than those from the LK2 group (p = 0.0002). In the LK0 group FITC-D leakage correlated significantly with the inflammation (p < 0.001).At the selected conditions, Visudyne^®-mediated PDT promotes vascular leakage and FITC-D extravasation into the interstitial space of normal tissue. The intensity of vascular leakage depends on the time interval between PDT and FITC-D injection. This concept could be used to locally modulate the delivery of macromolecules in vivo.     

Positron annihilation study of proton-irradiated reactor pressure vessel steels

The microstructures, irradiation-induced defects and changes of mechanical property of Chinese domestic A508-3 steels after proton irradiation were investigated by TEM, positron lifetime, slow positron beam Doppler broadening spectroscopy and hardness measurements. The defects were induced by 240 keV proton irradiation with fluences of 1.25×10¹⁷ ions cm^?2 (0.26 dpa), 2.5×10¹⁷ ions cm^?2 (0.5 dpa), and 5.0×10¹⁷ ions cm^?2 (1.0 dpa). The TEM observation revealed that the as-received steel had typical bainitic–ferritic microstructures. It was also observed that Doppler broadening S-parameter and average lifetime increased with dose level owing to the formation of defects and voids induced by proton irradiation. The correlation between positron parameters and hardness was found.     

Microstructural modifications in swift ion irradiated PET

Polyethylene terephthalte (PET) was irradiated with carbon (70 MeV) and copper (120 MeV) ions to analyze the induced modifications with respect to optical, structural and thermal properties. In the present investigation, the fluence for carbon irradiation was varied from 1×10¹¹ to 1×10¹⁴ ions cm⁻², while that for copper beam was kept in the range of 1×10¹¹ to 1×10¹³ ions cm⁻². UV–vis, FTIR, XRD and DSC techniques were utilized to study the induced changes. The analysis of UV–vis absorption studies reveals that there is decrease of optical energy gap up to 10% on carbon ion irradiation (at 1×10¹⁴ ions cm⁻²), whereas the copper beam (at 1×10¹³ ions cm⁻²) leads to a decrease of 49%. FTIR analysis indicated the formation of alkyne end groups along with the overall degradation of polymer with copper ion irradiation. X-ray diffraction analysis revealed that the semi-crystalline PET losses its crystallinity on swift ion irradiation. It was found that the carbon beam (1×10¹⁴ ions cm⁻²) decreased the crystallite size by 16% whereas this decrease is of 12% in case of the copper ion irradiated PET at 1×10¹³ ions cm⁻². The loss in crystallinity on irradiation has been supported by DSC thermograms.     

Electronic and vibrational spectra of tourmaline – The impact of electron beam irradiation and heat treatment

Irradiation and heat treatment were performed on tourmalines of various colors from Antandrokomby, Madagascar. The samples were irradiated with 10 MeV electrons to fluencies of 2 ×10¹⁷ cm⁻² for 1 h and were heated at 550 °C for 3 h in air. Their electronic and vibrational spectra were investigated by UV–vis, mid-infrared, and WD-XRF spectroscopy for comparison to pristine samples. Changes in the Mn³⁺ ions after irradiation resulted in darker pink tourmalines, which had absorption peaks at 390 and 520 nm. These samples became colorless after subsequent heat treatment. After irradiation, colorless, light blue and yellow tourmalines displayed a new absorption band at 365 nm. Alteration of the stretching absorption bands and wavenumber after irradiation could be explained by the following reactions:OH⁻ + e⁻ beam irradiation → O⁻ + H°,Mn²⁺ + e⁻ beam irradiation → Mn³⁺ + e⁻ andFe²⁺ + e⁻ beam irradiation → Fe³⁺ + e⁻.Stretching vibration of the BO₃ structure occurred at 1330 cm⁻¹, while the SiO vibration absorption bands were assigned to around 1100 cm⁻¹. Colorless, green, and yellow tourmalines showed high-intensity peaks around 3608 and 3505 cm⁻¹ after irradiation. Pink and dark green tourmalines showed low-intensity peaks at 3605 and 3585 cm⁻¹, respectively. The combination modes of stretching and bending in the range of 4600–4300 cm⁻¹ were split after irradiation and heat treatment, and different color changes occurred after irradiation.     

Proton (3 MeV) and copper (120 MeV) ion irradiation effects in low-density polyethylene (LDPE)

Low-density polyethylene (LDPE) was irradiated with proton (3 MeV) and copper (120 MeV) ions to analyze the induced modifications with respect to optical and structural properties. In the present investigation, the fluence for proton irradiation was varied up to 2×10¹⁵ protons cm⁻², while that for copper beam was kept in the range of 1×10¹ to 1×10¹³ ions cm⁻² to study the swift heavy ion-induced modifications in LDPE. Ultraviolet–visible (UV–vis), FTIR and X-ray diffraction (XRD) techniques were utilized to study the induced changes. The analysis of UV–vis absorption studies reveals that there is decrease of optical energy gap up to 43% on proton irradiation (at 2×10¹⁵ ions cm⁻²), whereas the copper beam (at 1×10¹³ ions cm⁻²) leads to a decrease of 51%. FTIR analysis indicated the presence of unsaturations due to vinyl end groups in the irradiated sample. The formation of OH and CO groups has also been observed. XRD analysis revealed that the semi-crystalline LDPE losses its crystallinity on swift ion irradiation. It was found that the proton beam (2×10¹⁵ ions cm⁻²) decreased the crystallite size by 23% whereas this decrease is of 31% in case of the copper ion-irradiated LDPE at 1×10¹³ ions cm⁻².     

Radiation induced in-situ generation of conductivity in the blends of polyaniline-base with chlorinated-polyisoprene

Radiation induced acid doping of PANI to generate electrical conductivity was achieved by radiation induced HCl release from chlorinated-polyisoprene (ClPIP). Blends of PANI with ClPIP were prepared by mechanical mixing/grinding in the composition range of 9–43% ClPIP by weight and pelletized under 10 t press. The pellets were irradiated in ⁶⁰Co Gammacell in air at room temperature to doses up to 300 kGy. The maximum electrical conductivity increase was observed for the blend PANI43 which changed from 10^?10 to 10^?4 S cm^?1 when it was irradiated to 300 kGy dose. Radiation induced changes on the blends were also studied by UV–vis spectroscopy using reflection technique and FTIR spectroscopy. The broad absorption band in the visible range (630 nm) increased by increasing irradiation dose. The band (1110 cm^?1) in the IR spectra which is indicative of conductivity showed linear correlation with irradiation dose.     

Application of non-aggressive sample preparation and electrothermal atomic absorption spectrometry to quantify platinum in biological matrices after cisplatin nanoparticle administration

A reproducible, simple, rapid and sensitive electrothermal atomic absorption spectrometry (ETAAS) method using low volume of sample (< 50 μL) was described for the quantitative analysis of cis-diamminedichloroplatinum (II) (cisplatin) in several biological matrices after PLGA [Poly (d,l-lactic-co-glycolic acid)] cisplatin nanoparticles (NP) administration to tumor-bearing mice. The validation parameters such as specificity, linearity, accuracy and precision obtained in this method were according to FDA guidelines and are presented in the text, as well as the limit of detection and quantification. Two applications were carried out with this method. In-vitro application was used to measure the loading efficacy of cisplatin in the formulation, and to characterize the drug kinetic release in culture cell medium. For in-vivo application, three groups of nu/nu mice injected with tumor cells were treated with 5 mg kg^? 1 dose of free cisplatin by intravenous and intraperitoneal routes, respectively, and with NP by intraperitoneal route. Blood samples were collected at different times, to measure cisplatin plasma concentrations, and at the end of the study, different organs were removed from each animal to quantify drug distribution. Additionally, the relationship between cisplatin levels and its apoptotic activity in each tumor sample, could be also characterized. This simple and sensitive method without aggressive manipulation allowed the quantification of cisplatin in different biological matrices with independence of the formulation used. Therefore, these pre-clinical results show the possibility to include this method for clinical applications.     

Analysis and identification of irradiated Spirulina powder by a three-step infrared macro-fingerprint spectroscopy

A three-step infrared (IR) macro-fingerprint method combining conventional IR spectra, and the secondary derivative spectra with two-dimensional infrared correlation spectroscopy (2D-IR), was developed to analyze Spirulina powder before and after gamma irradiation. In the IR spectra, most of the absorption peaks of samples irradiated at 1, 2.7, 6, and 10.4 kGy had lower intensities than the non-irradiated ones, whereas peaks at 1152, 1078, and 1051 cm⁻¹ were slightly enhanced with irradiation at 2.7, 6, and 10.4 kGy. Their second derivative spectra amplified the differences and revealed that irradiation affected the C=O band of carboxylic acid and esters, and the N–H band of proteins. The peaks at 1746 and 1741 cm⁻¹, and those at 1730 and 1725  cm⁻¹ became two broad peaks. Meanwhile, the three sharp peaks at 1548 cm⁻¹, 1544 cm⁻¹ and 1536 cm⁻¹ changed to two broad peaks at around 1547 and 1534 cm⁻¹ after irradiation at doses higher than 1 kGy. The characteristic IR bands from 1700 cm⁻¹ to 1600 cm⁻¹, which represent the C=O band in proteins, also have different shapes and intensities after irradiation. The finding indicated that irradiation affected the secondary structures of protein which was confirmed by curve fitting results. During the process of increasing the temperature from 50 to 210 °C, the ratio of amide I to II in absorption intensities in the 2D-IR spectra of the irradiated samples varied with different response for different samples. Saccharides in Spirulina powder had a higher thermostability than proteins, but the autopeaks of irradiated samples did show differences from the non-irradiated sample. The intensity of autopeaks at 1012 cm⁻¹ increased dramatically in the irradiated samples while that of peaks at 1053, 1071, and 1083 cm⁻¹ decreased after irradiation. Based on the three-step IR macro-fingerprint method, irradiated Spirulina powder samples were successfully and fast identified and discriminated.     

Effect of swift heavy ion irradiation on ethylene–chlorotrifluoroethylene copolymer

The swift heavy irradiation induced changes taking place in ethylene–chlorotrifluoroethylene (E–CTFE) copolymer films were investigated in correlation with the applied doses. Samples were irradiated in vacuum at room temperature by lithium (50 MeV), carbon (85 MeV), nickel (120 MeV) and silver (120 MeV) ions with the fluence in the range of 1×10¹¹–3×10¹² ions cm^?2. Structural and thermal properties of the irradiated as well as pristine E–CTFE films were studied using FTIR, UV–visible, TGA, DSC and XRD techniques. Swift heavy ion irradiation was found to induce changes in E–CTFE depending upon the applied doses.     

Effect of Steeped Black Tea (Camellia sinensis (L.) var. assamica) on Immunoglobulin Titer and Lymphocyte Proliferation in Responses to Hepatitis B Vaccine in Mice

Effect of steeped black tea (Camellia sinensis (L.) var. assamica) on immunoglobulin titer (IgM and IgG) and lymphocyte proliferation in responses to hepatitis B vaccine inBALB/c mice has been investigated. The study was divided into two stages i.e. the determination of immunoglobulin titer and the test of lymphocytes proliferation. In the first stage mice were divided into 5 groups each consisting of 5 mice. Group I, II and III was given steeped black tea respectively with a dose of 600 mg/kg bw; 1.2 g/kg bw and 2.4 g/kg bw. Group IV was given Stimuno^® with a dose of 6.5 mg/kg bw, and group V was given aquadest as negative control. All groups were induced by hepatitis B vaccine on day-0 (after 7 days of acclimatization). Serum was taken on day-14 and 21 for measurement of IgM titer and IgG, respectively. In the second stage, the mice were grouped as in the first stage, then all groups were induced by hepatitis B vaccine at day-0 and day-7. On day-27 lymphocyte was isolated and then tested for the growth and proliferation of lymphocytes. The results of this study showed that the steeped black tea has an effect in increasingIgM and IgG titer of BALB/cmice induced by hepatitis B vaccine, where the most effective dose was 1.2 g/kg bw. Steeped black tea also could increase lymphocytes proliferation in mice BALB/cinduced by hepatitis B vaccine, where the most effective dose was 1.2 g/kg bw.     

Photocatalytic degradation of paraquat and genotoxicity of its intermediate products

The photocatalytic degradation of paraquat (1,1-dimethyl-4,4′-bipyridylium dichloride) aqueous solutions in the presence of polycrystalline TiO₂ Degussa P25 irradiated by near-UV light was investigated. The substrate and total organic carbon concentrations were monitored by UV spectroscopy and TOC measurements, respectively: the complete photocatalytic mineralization of paraquat (20 ppm) was achieved after ca. 3 h of irradiation by using 0.4 g l⁻¹ of catalyst amount at natural pH (ca 5.8). On the contrary no significant photodegradation of paraquat was observed in the absence of TiO₂ under similar experimental conditions. To evaluate the genotoxicity of paraquat and its intermediates produced during heterogeneous photocatalytic treatment, in vitro tests such as Ames test, with and without rat liver microsomal fractions (S9 mix), and micronucleus test, were used. Results obtained with Salmonella typhimurium (strain TA100) showed that paraquat and photocatalytic products were unable to induce gene mutations when photocatalysis was used in the presence of the optimum amount of TiO₂, i.e. 0.4 g l⁻¹, whereas an increase of revertants his⁺ per plate was observed after 300 min irradiation in the presence of very low amount of TiO₂ (0.04 g l⁻¹). The negative results from micronucleus test suggest that mutagenic, but non-clastogenic, late intermediates of paraquat photo-oxidation were formed when the photocatalytic runs of paraquat degradation were carried out by using 0.04 g l⁻¹ of photocatalyst.     

Effects of gamma and electron beam irradiation on the survival of pathogens inoculated into sliced and pizza cheeses

The objective of this study was to identify the efficacy of gamma and electron beam irradiation of the food-borne pathogens (Listeria monocytogenes and Staphylococcus aureus) in sliced and pizza cheeses commercially available in the Korean market. Total aerobic bacteria and yeast/mold in the cheeses ranged from 10² to 10³ Log CFU/g. Irradiation of 1 kGy for sliced cheese and 3 kGy for pizza cheese were sufficient to lower the total aerobic bacteria to undetectable levels (10¹ CFU/g). Pathogen inoculation test revealed that gamma irradiation was more effective than electron beam irradiation at the same absorbed dose, and the ranges of the D₁₀ values were from 0.84 to 0.93 kGy for L. monocytogenes and from 0.60 to 0.63 kGy for S. aureus. Results suggest that a low dose irradiation can improve significantly the microbial quality and reduce the risk of contamination of sliced and pizza cheeses by the food-borne pathogens which can potentially occur during processing.     

Temporal evolution study of the plasma induced by CO2 pulsed laser on targets of titanium oxides

This paper reports studies on time-resolved laser induced breakdown spectroscopy (LIBS) of plasmas induced by IR nanosecond laser pulses on the titanium oxides TiO and TiO₂ (anatase). LIBS excitation was performed using a CO₂ laser. The laser-induced plasma was found strongly ionized yielding Ti⁺, O⁺, Ti^2 +, O^2 +, Ti^3 +, and Ti^4 + species and rich in neutral titanium and oxygen atoms. The temporal behavior of specific emission lines of Ti, Ti⁺, Ti^2 + and Ti^3 + was characterized. The results show a faster decay of Ti^3 + and Ti^2 + ionic species than that of Ti⁺ and neutral Ti atoms. Spectroscopic diagnostics were used to determine the time-resolved electron density and excitation temperatures. Laser irradiation of TiO₂-anatase induces on the surface sample the polymorphic transformation to TiO₂-rutile. The dependence on fluence and number of irradiation pulses of this transformation was studied by micro-Raman spectroscopy.     

In vivo anticancer and histopathology studies of Schiff bases on Ehrlich ascitic carcinoma cells: 1st Cancer Update

Three Schiff bases in two different concentrations were evaluated for their anti-tumor activity against Ehrlich ascites carcinoma (EAC) bearing Swiss albino mice. The in vivo anti-tumor potency of Schiff bases was assessed by measuring the increase in mean survival time of the drug treated over untreated control mice and treated standard (cisplatin) mice. Their toxicity was assessed in vivo in normal, standard, and EAC-bearing mice by measuring the drug-induced changes in biochemical as well as hematological parameters. The histopathology studies to assess the toxicity of these compounds on vital organs also have been studied. Among the three Schiff bases studied, 4-({[3-(4-fluorophenyl)-1H-pyrazol-4-yl]methylene}amino)-5-[(2-methylphenoxy)methyl]-1,2,4-triazole-3-thiol (SB-3) at an optimal dose of 100 mg/kg body weight was found to enhance the mean survival time of infected mice. Deviated hematological parameters and mean survival time in tumor bearing mice were found to be significantly restored towards normal after treatment with SB-3 100 mg/kg body weight of mice. The ALP and SGOT values were found to approach the normal range. A:G ratios also did not deviate from normal on treatment with SB-3. The histopathology studies revealed only mild hepatotoxicity and nephrotoxicity when compared to the normal and standard. The splenic cellularity also did not show much variation from normal. SB-3 at a prime dose of 100 mg has shown promising anticancer activity in vivo against EAC when compared to standard drug with minimum toxic effects.     

Structural modifications of swift heavy ion irradiated PEN probed by optical and thermal measurements

The effects of swift heavy ion irradiation on the structural characteristics of Polyethylene naphthalate (PEN) were studied. Samples were irradiated in vacuum at room temperature by lithium (50 MeV), carbon (85 MeV), nickel (120 MeV) and silver (120 MeV) ions with the fluence in the range of 1×10¹¹–3×10¹²  ions cm⁻². Ion induced changes were analyzed using X-ray diffraction (XRD), Fourier transform infra red (FT-IR), UV–visible spectroscopy, thermo-gravimetric analysis (TGA) and differential scanning calorimetry (DSC) techniques. Cross-linking was observed at lower doses resulting in modification of structural properties, however higher doses lead to the degradation of the investigated polymeric samples.     

Reinforcement of natural rubber/high density polyethylene blends with electron beam irradiated liquid natural rubber-coated rice husk

Coating of rice husk (RH) surface with liquid natural rubber (LNR) and exposure to electron beam irradiation in air were studied. FTIR analysis on the LNR-coated RH (RHR) exposed to electron beam (EB) showed a decrease in the double bonds and an increase in hydroxyl and hydrogen bonded carbonyl groups arising from the chemical interaction between the active groups on RH surface with LNR. The scanning electron micrograph showed that the LNR formed a coating on the RH particles which transformed to a fine and clear fibrous layer at 20 kGy irradiation. The LNR film appeared as patches at 50 kGy irradiation due to degradation of rubber. Composites of natural rubber (NR)/high density polyethylene (HDPE)/RHR showed an optimum at 20–30 kGy dosage with the maximum stress, tensile modulus and impact strength of 6.5, 79 and 13.2 kJ/m², respectively. The interfacial interaction between the modified RH and TPNR matrix had improved on exposure of RHR to e-beam at 20–30 kGy dosage.     

Effect of high-dose irradiation and autoclave treatment on microbial safety and quality of ready-to-eat Bulgogi sauce

In Korea, commercialized sauce for ready-to-eat (RTE) Bulgogi is usually manufactured using heat treatment to ensure that it has a long shelf-life. However, heat treatment may adversely affect the taste and flavor of the sauce, thus, the development of suitable sterilizing methods for RTE sauces is necessary to preserve the quality of the sauce during long storage periods. In this study, total bacterial growth, the viscosity, and the sensory properties of Bulgogi sauce were compared between sterilization with gamma irradiation (0–40 kGy) and autoclave treatment during storage at 35 °C for 90 days. No bacterial growth was observed following irradiation at more than 10 kGy or after autoclave treatment. However, the viscosity and sensory properties of samples gamma-irradiated at above 10 kGy or autoclave-treated were significantly changed, even though autoclave treatment induced a burnt taste and flavor. Therefore, a gamma irradiation of 10 kGy was effective to prepare ready-to-eat Bulgogi sauce with microbial safety and original sensory qualities.     

Radiolytic stabilization of poly(hydroxybutyrate)

Stabilization against gamma irradiation effects on Brazilian poly(hydroxybutyrate) (PHB) films was investigated by using commercial additives. With the most effective additive the G value of main chain scissions decreased from 8.6 to 1.5 scissions/100 eV in the absorbed dose range up to 35 kGy. Irradiation increased the biodegradability of the samples, whereas the additives-induced reduction in biodegradability. Irradiation treatment did not influence the degree of crystallinity. Protonic nuclear magnetic resonance (NMR-¹H) analysis revealed formation of hydroxyl-terminal groups upon irradiation.     

Combined effect of γ-irradiation and bacterial-fermented dextrose on microbiological quality of refrigerated pork sausages

The objective of this study was to evaluate the effect of a concentrated fermented dextrose (FD), a natural antimicrobial product, combined with low dose γ-irradiation (1.5 kGy) on the microbiological quality of fresh pork sausages. Fresh pork sausages containing the FD (0.25%, 0.5% and 0.75%) were prepared in a meat pilot plant and were irradiated using a UC-15A irradiator equipped with a ⁶⁰Cobalt source. The γ-irradiation treatment alone was able to reduce the initial psychrophilic and mesophilic bacteria by more than 2 log CFU/g and kept the lactobacillus population under the detection limit (100 CFU/g). Results also showed that the FD alone was able to extend the shelf life of the sausages from 5 days up to 13 days. At day 13, the FD or irradiation alone showed 2 log CFU/g less mesophilic bacteria than the control. After combining FD and irradiation another reduction of the microbial count of 1 log CFU/g was observed. When combining the irradiation treatment with the FD results it showed a reduced growth rate of the psychrophilic and mesophilic bacteria compared to both treatments alone. This study demonstrated that FD with low dose gamma irradiation act in synergy to reduce the multiplication of the total bacterial flora in fresh sausages.