Application of Spectroscopic Methods for the Identification of Superoxide Dismutases in Cyanobacteria.

Superoxide dismutases (SODs) belong to the group of metalloenzymes that remove superoxide anion radicals and they have been identified in three domains of life: Bacteria, Archaea and Eucarya. SODs in Synechocystis sp. PCC 6803, Gloeobacter violaceus CCALA 979, and Geitlerinema sp. ZHR1A were investigated. We hypothesized that iron (FeSOD) and/or manganese (MnSOD) dominate as active forms in these cyanobacteria. Activity staining and three different spectroscopic methods of SOD activity bands excised from the gels were used to identify a suitable metal in the separated samples. FeSODs or enzymes belonging to the Fe-MnSOD superfamily were detected. The spectroscopic analyses showed that only Fe is present in the SOD activity bands. We found FeSOD in Synechocystis sp. PCC 6803 while two forms in G. violaceus and Geitlerinema sp. ZHR1A: FeSOD1 and FeSOD2 were present. However, no active Cu/ZnSODs were identified in G. violaceus and Geitlerinema sp. ZHR1A. We have shown that selected spectroscopic techniques can be complementary to the commonly used method of staining for SOD activity in a gel. Furthermore, the occurrence of active SODs in the cyanobacteria studied is also discussed in the context of SOD evolution in oxyphotrophs.

The Influence of Biostimulants Used in Sustainable Agriculture for Antifungal Protection on the Chemical Composition of Winter Wheat Grain.

Field studies were conducted from 2016 to 2019 (south-eastern Poland; 49°58'40.6″ N 22°33'11.3″ E) with the aim to identify the chemical composition of winter wheat grain upon foliar application of biostimulants, of which PlanTonic BIO (containing nettle and willow extracts) showed antifungal activity. The main chemical compositions and their spatial distribution in wheat grain were characterized by Raman spectroscopy technique. It was established that applied biostimulants and hydro-thermal conditions changed the chemical composition of the grain during all the studied years. A similar chemical composition of the grain was achieved in plants treated with synthetic preparations, including both intensive and extensive variants. The second group, in terms of an increase in fatty acid content, consists of grains of plants treated with biostimulants PlanTonic BIO, PlanTonic BIO + Natural Crop and PlanTonic BIO + Biofol Plex. The future of using biostimulants in crop production, including those containing salicylic acid and nettle extracts, appears to be a promising alternative to synthetic crop protection products.

Spectrochemical and biochemical assay comparison study of the healing effect of the Aloe vera and Hypericum perforatum loaded nanofiber dressings on diabetic wound.

Diabetic wounds have a slow healing process and easy to be infected. In addition to current drug treatments, supportive approaches are needed for diabetic wound treatment. In this study, we aimed to load Aloe Vera (AV) and Hypericum perforatum oil (HPO) with PCL/Ge (Poly (ɛ-caprolactone)/Gelatine) polymeric biodegradable by electrospinning method into nanofiber dressings on an experimental diabetic wound model to compare the diabetic wound healing effect. Changes in the amount and chemical structure of phospholipids, proteins, and lipids were investigated in the blood and serum samples of the animals using Fourier transform infrared (FTIR) analysis. To evaluate biological events associated with the wound repair process in inflammatory phase we used oxidant and antioxidant status to determine the healing status of wounds such as Total antioxidant status (TAS), Total oxidant level (TOS) and tumor necrosis factor alpha (TNF-α) levels. TOS level increased in DM groups and decreased in the AV and HPO group. Oxidative stress index decreased and TNF-α level increased in the HPO group. FTIR spectra showed changes in the phospholipids, proteins, and carbon chain of lipids in the whole blood as well as serum of DM rats. FTIR spectra combined with Principal component analysis (PCA) showed, that treated DM rats by AV and HPO caused return chemical structure of blood and serum to this observed in control group. Higher similarity with control group for HPO rats was observed. HPO is better than AV in the alternative for healing on diabetic wound. Thus, we have demonstrated that IR spectroscopy and multivariate data analysis and biochemical assays are consistent and correlative with each other.

Treating honey bees with an extremely low frequency electromagnetic field and pesticides: Impact on the rate of disappearance of azoxystrobin and λ-cyhalothrin and the structure of some functional groups of the probabilistic molecules.

The purpose of these laboratory tests was to assess the impact of 50 Hz EMF (electromagnetic field) on the disappearance of azoxystrobin (active ingredient (AI) of Amistar 250 SC) and λ-cyhalothrin (AI of Karate Zeon 050 CS) in the body of honey bees (Apis mellifera) and the structure of some functional groups of the probabilistic molecules in their organisms. Amistar 250 SC (an azoxystrobin-based fungicide; ABF) and Karate Zeon 050 CS (a λ-cyhalothrin-based insecticide; CBI) are plant protection products (PPPs) applied to bee-pollinated-crops. Chromatographic methods were used to assess the rate of AI disappearance. EMF affected the rate of disappearance of azoxystrobin and λ-cyhalothrin in bees within 6 h of intoxication. When these substances were used separately their disappearance in the presence of EMF slowed from 12.6% to 10.5% h-1 and from 9.2% to 4.8% h-1, respectively, and accelerated when used in a mixture, from 14.1% to 14.7% h-1 and from 9.3% to 11.5% h-1 respectively. Fourier Transform Infrared (FTIR) spectroscopy was used to analyze changes in the functional groups of the probabilistic molecules of the tested bees. To obtain the information about the spectra variations we used the Principal Component Analysis. It has been shown, that EMF statistically significantly interferes with amide I and II, symmetric PO32- group from DNA, RNA and phospholipids vibrations. It also increased the number of changes of functional groups of the probabilistic molecules caused by ABF, but at the same time limited the changes in the functional groups studied in bees treated with CBI and a mixture containing both of them. In addition, exposure to EMF in bees treated with fungicide or insecticide, separately, and with both preparations caused differences (p < 0.05) in the secondary structure of proteins compared to controls. The obtained results indicate that EMF may affect the rate of metabolism and the detoxification process of pesticides in bees, depending on the AI of PPPs, applied individually or together. However, further detailed research is required to explain the mechanism of EMF as a detoxification modulator.

Control of Arms of Au Stars Size and its Dependent Cytotoxicity and Photosensitizer Effects in Photothermal Anticancer Therapy.

Gold nanostars (AuS NPs) are a very attractive nanomaterial, which is characterized by high effective transduction of the electromagnetic radiation into heat energy. Therefore, AuS NPs can be used as photosensitizers in photothermal therapy (PTT). However, understanding the photothermal conversion efficiency in nanostars is very important to select the most appropriate shape and size of AuS NPs. Therefore, in this article, the synthesis of AuS NPs with different lengths of star arms for potential application in PTT was investigated. Moreover, the formation mechanism of these AuS NPs depending on the reducer concentration is proposed. Transmission electron microscopy (TEM) with selected area diffraction (SEAD) and X-ray diffraction (X-Ray) showed that all the obtained AuS NPs are crystalline and have cores with similar values of the diagonal (parameter d), from 140 nm to 146 nm. However, the widths of the star arm edges (parameter c) and the lengths of the arms (parameter a) vary between 3.75 nm and 193 nm for AuS1 NPs to 6.25 nm and 356 nm for AuS4 NPs. Ultraviolet-visible (UV-Vis) spectra revealed that, with increasing edge widths and lengths of the star arms, the surface plasmon resonance (SPR) peak is shifted to the higher wavelengths, from 640 nm for AuS1 NPs to 770 nm for AuS4 NPs. Moreover, the increase of temperature in the AuS NPs solutions as well as the values of calculated photothermal efficiency grew with the elongation of the star arms. The potential application of AuS NPs in the PTT showed that the highest decrease of viability, around 75%, of cells cultured with AuS NPs and irradiated by lasers was noticed for AuS4 NPs with the longest arms, while the smallest changes were visible for gold nanostars with the shortest arms. The present study shows that photothermal properties of AuS NPs depend on edge widths and lengths of the star arms and the values of photothermal efficiency are higher with the increase of the arm lengths, which is correlated with the reducer concentration.