Budbreak date in grapevine is strictly dependent on temperature, and the correct simulation of its occurrence is of great interest since it may have major consequences on the final yield and quality. In this study, we evaluated the reliability for budbreak simulation of two modeling approaches, the chilling-forcing (CF), which describes the entire dormancy period (endo- and eco-dormancy) and the forcing approach (F), which only describes the eco-dormancy. For this, we selected six phenological models that apply CF and F in different ways, which were tested on budbreak simulation of eight grapevine varieties cultivated at different latitudes in Europe. Although none of the compared models showed a clear supremacy over the others, models based on CF showed a generally higher estimation accuracy than F where fixed starting dates were adopted. In the latter models, the accurate simulation of budbreak was dependent on the selection of the starting date for forcing accumulation that changes according to the latitude, whereas CF models were independent. Indeed, distinct thermal requirements were found for the grapevine varieties cultivated in Northern and Southern Europe. This implies the need to improve modeling of the dormancy period to avoid under- or over-estimations of budbreak date under different environmental conditions. Full article

The obligate biotroph Plasmodiophora brassicae causes clubroot disease in oilseeds and vegetables of the Brassicaceae family, and cytokinins play a vital role in clubroot formation. In this study, we examined the expression patterns of 17 cytokinin-related genes involved in the biosynthesis, signaling, and degradation in Chinese cabbage inoculated with the Korean pathotype group 4 isolate of P. brassicae, Seosan. This isolate produced the most severe clubroot symptoms in Chinese cabbage cultivar “Bullam-3-ho” compared to three other Korean geographical isolates investigated. BrIPT1, a cytokinin biosynthesis gene, was induced on Day 1 and Day 28 in infected root tissues and the upregulation of this biosynthetic gene coincided with the higher expression of the response regulators BrRR1, on both Days and BrRR6 on Day 1 and 3. BrRR3 and 4 genes were also induced during gall enlargement on Day 35 in leaf tissues. The BrRR4 gene, which positively interact with phytochrome B, was consistently induced in leaf tissues on Day 1, 3, and 14 in the inoculated plants. The cytokinin degrading gene BrCKX3-6 were induced on Day 14, before gall initiation. BrCKX2,3,6 were induced until Day 28 and their expression was downregulated on Day 35. This insight improves our current understanding of the role of cytokinin signaling genes in clubroot disease development. Full article

The effect of doping the ferrite alloy Fe-16Cr by the oversized impurities Sb and Au on the mechanism of the short-range ordering induced by “warm” severe plastic deformation was studied using the method of Mössbauer spectroscopy. A comparison between the results obtained and the positron annihilation data on the evolution of the defects of vacancy type stabilized by the impurities Sb and Au was performed. It has been established that the impurities Sb and Au entail a shift of the temperature region of short-range ordering realization in conditions of applying pressure torsion towards greater temperatures by 250 and 100 К, respectively. Full article

In patients with severe heart failure, the surgical reconstruction of the damaged myocardium with regenerative biological grafts is an innovative therapeutic option. However, natural patch materials are often too delicate for a full wall repair of the left ventricle. A degradable magnesium scaffold could provide temporary mechanical stability until the sufficient physiological remodeling of such grafts. An autologous vascularised gastric patch was employed for the reconstruction of the left ventricular myocardium in a porcine model. Magnesium alloy (LA63) scaffolds were fixed over the biological patch. The function of the implant was assessed via magnetic resonance imaging. Angiography was carried out to detect a connection between the gastric and coronary vasculature. The explants were examined via µ-computer tomography and light microscopy. All the test animals survived. The prostheses integrated biologically and functionally into the myocardium. No rupture of the prostheses occurred. An anastomosis of the gastric and coronary vasculature had developed. The magnesium scaffolds degraded, on average, to 30.9% of their original volume. This novel technique responds to the increasing demand for regenerative myocardial grafts. The magnesium scaffolds’ biocompatibility and degradation kinetics, as well as their stabilizing effects, indicate their applicability in the surgical treatment of terminal heart failure. Full article

Previous studies revealed the potential of Labrenzia aggregata USBA 371 to produce cytotoxic metabolites. This study explores its metabolic diversity and compounds involved in its cytotoxic activity. Extracts from the extracellular fraction of strain USBA 371 showed high levels of cytotoxic activity associated with the production of diketopiperazines (DKPs). We purified two compounds and a mixture of two other compounds from this fraction. Their structures were characterized by 1D and 2D nuclear magnetic resonance (NMR). The purified compounds were evaluated for additional cytotoxic activities. Compound 1 (cyclo (l-Pro-l-Tyr)) showed cytotoxicity to the following cancer cell lines: breast cancer 4T1 (IC₅₀ 57.09 ± 2.11 µM), 4T1H17 (IC₅₀ 40.38 ± 1.94), MCF-7 (IC₅₀ 87.74 ± 2.32 µM), murine melanoma B16 (IC₅₀ 80.87 ± 3.67), human uterus sarcoma MES-SA/Dx5 P-pg (−) (IC50 291.32 ± 5.64) and MES-SA/Dx5 P-pg (+) (IC₅₀ 225.28 ± 1.23), and murine colon MCA 38 (IC50 29.85 ± 1.55). In order to elucidate the biosynthetic route of the production of DKPs and other secondary metabolites, we sequenced the genome of L. aggregata USBA 371. We found no evidence for biosynthetic pathways associated with cyclodipeptide synthases (CDPSs) or non-ribosomal peptides (NRPS), but based on proteogenomic analysis we suggest that they are produced by proteolytic enzymes. This is the first report in which the cytotoxic effect of cyclo (l-Pro-l-Tyr) produced by an organism of the genus Labrenzia has been evaluated against several cancer cell lines. Full article

A polystyrene (PS)-based fibrous color dosimeter, comprising a color former based on 2-(phenylamino)-6-(dipentylamino)-3-methylspiro[9H-xanthene-9,3′-phthalide] (Black305) fluoran leuco dye and a 2-(4-methoxystyryl)-4,6-bis(trichloromethyl)-1,3,5-triazine (MBTT) photoacid generator, was developed for visual detection of X-ray doses of 15 Gy and higher. The composite fiber was produced by using a centrifugal spinning method, and the obtained composite fiber exhibited a stable and uniform morphology with a fiber diameter of 10 μm or less and had sufficient mechanical strength. As an example of practical application, we successfully processed the composite fiber into an apron and clearly and visually confirmed that the color change from yellow to black occurs on the surface of the fabric under X-ray exposure. Full article

A “Well-to-Propeller” Life Cycle Assessment of maritime transport was performed with a European geographical focus. Four typical types of vessels with specific operational profiles were assessed: a container vessel and a tanker (both with 2-stroke engines), a passenger roll-on/roll-off (Ro-Pax) and a cruise vessel (both with 4-stroke engines). All main engines were dual fuel operated with Heavy Fuel Oil (HFO) or Liquefied Natural Gas (LNG). Alternative onshore and offshore fuel supply chains were considered. Primary energy use and greenhouse gas emissions were assessed. Raw material extraction was found to be the most impactful life cycle stage (~90% of total energy use). Regarding greenhouse gases, liquefaction was the key issue. When transitioning from HFO to LNG, the systems were mainly influenced by a reduction in cargo capacity due to bunkering requirements and methane slip, which depends on the fuel supply chain (onshore has 64% more slip than offshore) and the engine type (4-stroke engines have 20% more slip than 2-stroke engines). The combination of alternative fuel supply chains and specific operational profiles allowed for a complete system assessment. The results demonstrated that multiple opposing drivers affect the environmental performance of maritime transport, a useful insight towards establishing emission abatement strategies. Full article

The majority of municipal Wastewater Treatment Plants (WWTPs) in Sweden produce biogas from sewage sludge. In order to increase the methane production, co-digestion of internal sludge with Organic Fraction of Municipal Solid Waste (OFMSW) might be feasible in the future. The objective of this study was therefore to find a beneficial solution for the utilization of OFMSW at the WWTP in Varberg, Sweden. The effects of co-digesting primary sludge (PS) and OFMSW collected in the municipality, in different mixing ratios, were investigated by semi-continuous anaerobic digestion assays. Furthermore, the effects of the addition of a commercial trace elements mixture solution (CTES), available on the market in Sweden, were also examined. Co-digestion of OFMSW and PS resulted in specific methane yields of 404, 392, and 375 Nml CH₄/g volatile solids (VS), obtained during semi-continuous operations of 301, 357 and 385 days, for the reactors fed with OMFSW:PS ratio of 4:1, 3:1, and 1:1, and at maximum organic loading rates (OLRs) achieved of 4.0, 4.0 and 5.0 gVS/L/d, respectively. Furthermore, mono-digestion of OFMSW failed already at OLR of 1.0 gVS/L/d, however, an OLR of 4.0 gVS/L/d could be achieved with addition of 14 µL/g VS Commercial Trace Element Solutions (CTES) leading to 363 mL CH₄/g VS methane production. These experiments were running during 411 days. Hence, higher process efficiency was obtained when using co-digestion of OFMSW and PS compared to that of OFMSW in mono-digestion. Co-digestion is a more feasible option where a balanced Carbon/Nitrogen (C/N) ratio and nutrient supply can be maintained. Full article

Intracellular concentration of reactive oxygen species (e.g., H₂O₂) in cancer cells is elevated over 10-fold as compared to normal cells. This feature has been used by us and several other research groups to design cancer specific prodrugs, for example, N-alkylaminoferrocene (NAAF)-based prodrugs. Further improvement of the efficacy of these prodrugs can be achieved by their targeting to intracellular organelles containing elevated reactive oxygen species (ROS) amounts. For example, we have previously demonstrated that lysosome-targeted NAAF-prodrugs exhibit higher anticancer activity in cell cultures, in primary cells and in vivo (Angew. Chem. Int. Ed. 2017, 56, 15545). Mitochondrion is an organelle, where electrons can leak from the respiratory chain. These electrons can combine with O₂, generating O₂^−• that is followed by dismutation with the formation of H₂O₂. Thus, ROS can be generated in excess in mitochondria and targeting of ROS-sensitive prodrugs to these organelles could be a sensible possibility for enhancing their efficacy. We have previously reported on NAAF-prodrugs, which after their activation in cells, are accumulated in mitochondria (Angew. Chem. Int. Ed. 2018, 57, 11943). Now we prepared two hybrid NAAF-prodrugs directly accumulated in mitochondria and activated in these organelles. We studied their anticancer activity and mode of action. Based on these data, we concluded that ROS produced by mitochondria is not available in sufficient quantities for activation of the ROS-responsive prodrugs. The reason for this can be efficient scavenging of ROS by antioxidants. Our data are important for the understanding of the mechanism of action of ROS-activatable prodrugs and will facilitate their further development. Full article

To avoid the lacquerware of the Nanhai No. 1 shipwreck from being corroded by microorganisms and to improve the knowledge on microbial ecology of the wood lacquers, we conducted a series of tests on the two water samples storing the lacquerware and colonies on the surface of the lacquerware. The high-throughput sequencing detected dominant fungal communities. After that, the fungal strains were isolated and then identified by amplification of ITS- 18S rRNA. Then the activity of ligninolytic and cellulolytic enzymes was detected on potato dextrose agar (PDA) plates with 0.04% (v/v) guaiacol and carboxymethyl cellulose (CMC) agar plates. Finally, we tested the biocide susceptibility of these fungi. Penicillium chrysogenum (NK-NH3) and Fusarium solani (NK- NH1) were the dominant fungi in the sample collected in April 2016 and June 2017. What is more, both showed activity of ligninolytic and cellulolytic enzymes. Four biocidal products (Preventol^® D7, P91, BIT 20N, and Euxyl^® K100) inhibited the growth of the fungal species in vitro effectively. In further research, the microbial community and environmental parameters in the museum should be monitored to assess the changes in the community and to detect potential microbial outbreaks. Full article

Survival rates for Ewing sarcoma (ES) patients with metastatic disease have not improved in over 20 years. Tumor growth and metastasis are dependent on tumor vasculature expansion; therefore, identifying the regulators that control this process in ES may provide new therapeutic opportunities. ES expresses high levels of repressor element 1 silencing transcription factor (REST), which is regulated by the EWS-FLI-1 fusion gene. However, the role of REST in ES growth and the regulation of the tumor vasculature have not been elucidated. To study this role, we established REST-knockout human TC71 ES cell lines through CRISPR/Cas9 recombination. While knockout of REST did not alter tumor cell proliferation in vitro, REST knockout reduced tumor growth and metastasis to the lung in vivo and altered tumor vascular morphology and function. Tumor vessels in the REST-knockout tumors had a punctate appearance with significantly decreased tumor vascular pericytes, decreased perfusion, and increased permeability. REST-knockout tumors also showed increased apoptosis and hypoxia. These results indicate that REST plays a critical role in ES vascular function, which in turn impacts the ability of ES tumors to grow and metastasize. These findings therefore provide a basis for the targeting of REST as a novel therapeutic approach in ES. Full article

Colorectal cancer (CRC) is the third most common cancer worldwide and the leading cause of cancer-related deaths. Recently, several studies have demonstrated that gut microbiota can alter CRC susceptibility and progression by modulating mechanisms such as inflammation and DNA damage, and by producing metabolites involved in tumor progression or suppression. Dysbiosis of gut microbiota has been observed in patients with CRC, with a decrease in commensal bacterial species (butyrate-producing bacteria) and an enrichment of detrimental bacterial populations (pro-inflammatory opportunistic pathogens). CRC is characterized by altered production of bacterial metabolites directly involved in cancer metabolism including short-chain fatty acids and polyamines. Emerging evidence suggests that diet has an important impact on the risk of CRC development. The intake of high-fiber diets and the supplementation of diet with polyunsaturated fatty acids, polyphenols and probiotics, which are known to regulate gut microbiota, could be not only a potential mechanism for the reduction of CRC risk in a primary prevention setting, but may also be important to enhance the response to cancer therapy when used as adjuvant to conventional treatment for CRC. Therefore, a personalized modulation of the pattern of gut microbiome by diet may be a promising approach to prevent the development and progression of CRC and to improve the efficacy of antitumoral therapy. Full article

Chitin is a structural polysaccharide of the cell walls of fungi and exoskeletons of insects and crustaceans. In this study, chitin was extracted, for the first time in our knowledge, from the Cicada orni sloughs of the south-eastern French Mediterranean basin by treatment with 1 M HCl for demineralization, 1 M NaOH for deproteinization, and 1% NaClO for decolorization. The different steps of extraction were investigated by Fourier Transform Infrared Spectroscopy (FTIR), X-Ray Diffraction (XRD), Thermogravimetric Analysis (TGA), and Scanning Electron Microscopy (SEM). Results demonstrated that the extraction process was efficiently performed and that Cicada orni sloughs of the south-eastern French Mediterranean basin have a high content of chitin (42.8%) in the α-form with a high degree of acetylation of 96% ± 3.4%. These results make Cicada orni of the south-eastern French Mediterranean basin a new and promising source of chitin. Furthermore, we showed that each step of the extraction present specific characteristics (for example FTIR and XRD spectra and, consequently, distinct absorbance peaks and values of crystallinity as well as defined values of maximum degradation temperatures identifiable by TGA analysis) that could be used to verify the effectiveness of the treatments, and could be favorably compared with other natural chitin sources. Full article

In order to investigate the effects of metallic nanoparticles (NPs) on the performance of in vitro bioassay, zinc oxide NP (ZnO NP), aluminum oxide NP (Al₂O₃ NP), bare silver NP (Ag NP), and Ag NP capped with citrate (Ag^cit NP) were evaluated with yeast (Saccharomyces cerevisiae Y190) two-hybrid system (YES assay), carrying Japanese medaka estrogen receptors (mERs) in the presence of 17β-estradiol (E2, 10⁻⁶ M), a reference chemical for estrogenic activity. The distribution of NPs in the yeast was also examined by field-emission transmission electron microscopy (FE-TEM). The results show that TEM analysis revealed that NPs were present inside the yeast and accumulated deep inside the cell organelles, suggesting that cell death was caused by NPs. However, despite no significant change of mortality, the E2 estrogenic activities in yeast exposed to ZnO NP and Al₂O₃ NP were dose-dependently reduced. For Ag NP and Ag^cit NP, such phenomenon observed in the exposure of ZnO NP and Al₂O₃ NP did not occur. From the observations, we found that ZnO NP and Al₂O₃ NP in the environmental media could result in underestimated estrogenicity of endocrine-disrupting compounds when evaluated by YES assay. Full article

Lipases are among the most utilized enzymes in biocatalysis. In many instances, the main reason for their use is their high specificity or selectivity. However, when full modification of a multifunctional and heterogeneous substrate is pursued, enzyme selectivity and specificity become a problem. This is the case of hydrolysis of oils and fats to produce free fatty acids or their alcoholysis to produce biodiesel, which can be considered cascade reactions. In these cases, to the original heterogeneity of the substrate, the presence of intermediate products, such as diglycerides or monoglycerides, can be an additional drawback. Using these heterogeneous substrates, enzyme specificity can promote that some substrates (initial substrates or intermediate products) may not be recognized as such (in the worst case scenario they may be acting as inhibitors) by the enzyme, causing yields and reaction rates to drop. To solve this situation, a mixture of lipases with different specificity, selectivity and differently affected by the reaction conditions can offer much better results than the use of a single lipase exhibiting a very high initial activity or even the best global reaction course. This mixture of lipases from different sources has been called “combilipases” and is becoming increasingly popular. They include the use of liquid lipase formulations or immobilized lipases. In some instances, the lipases have been coimmobilized. Some discussion is offered regarding the problems that this coimmobilization may give rise to, and some strategies to solve some of these problems are proposed. The use of combilipases in the future may be extended to other processes and enzymes. Full article

The concept of using ibuprofen as an albumin-binding entity was recently demonstrated by the development of [¹⁷⁷Lu]Lu-Ibu-PSMA-01. In the present study, we designed a novel ibuprofen-containing radioligand (Ibu-PSMA-02) with subtle structural changes regarding the linker entity in order to investigate a potential impact on the in vitro and in vivo properties. Ibu-PSMA-02 was prepared using solid-phase synthesis techniques and labeled with lutetium-177. [¹⁷⁷Lu]Lu-Ibu-PSMA-02 was evaluated in vitro with regard to its plasma protein-binding properties, PSMA affinity and uptake into PSMA-expressing PC-3 PIP tumor cells. The tissue distribution profile of [¹⁷⁷Lu]Lu-Ibu-PSMA-02 was assessed in tumor-bearing mice and dose estimations were performed. The in vitro characteristics of [¹⁷⁷Lu]Lu-Ibu-PSMA-02 were similar to those previously obtained for [¹⁷⁷Lu]Lu-Ibu-PSMA-01 with respect to plasma protein-binding, PSMA affinity and tumor cell uptake. The in vivo studies revealed, however, an unprecedentedly high uptake of [¹⁷⁷Lu]Lu-Ibu-PSMA-02 in PC-3 PIP tumors, resulting in an increased absorbed tumor dose of 7.7 Gy/MBq as compared to 5.1 Gy/MBq calculated for [¹⁷⁷Lu]Lu-Ibu-PSMA-01. As a consequence of the high tumor accumulation, [¹⁷⁷Lu]Lu-Ibu-PSMA-02 showed higher tumor-to-background ratios than [¹⁷⁷Lu]Lu-Ibu-PSMA-01. This study exemplified that smallest structural changes in the linker entity of PSMA radioligands may have a significant impact on their pharmacokinetic profiles and, thus, may be applied as a means for ligand design optimization. Full article