This work contributes to the issue of Cd content in Hypericum perforatum and Thymus serpyllum grown in localities of Rtanj and Ozren, and implies the importance for systemic control of Cd content in Hypericum species in order to provide safety of their preparations. Furthermore, regarding Cd toxicity, maximal permissible level of Cd in plant material should be evaluated and established concerning national legislative frame.

Background: Understanding the molecular features associated with response to neoadjuvant chemoradiotherapy is an unmet clinical need in locally advanced rectal cancer (LARC). The aim of the study was to apply a high-sensitivity proteomic approach for in-depth characterization of the LARC proteome in search of patients who might have a good response to preoperative treatment and potentially be followed by a watch-and-wait strategy, rather than having immediate surgery, maximizing the therapeutic effect and quality of life. Methods: A total of 97 LARC patients treated at the Institute for Oncology and Radiology of Serbia in the period of 2018-2019 were included in the study. Patients were treated with long-course chemoradiotherapy (CRT): Radiotherapy (RT) was delivered with a total dose of 50.4 Gy in 28 fractions; concomitant chemotherapy (5-FU, 350 mg/m2 daily) and Leucovorin (25 mg/m2 daily) was administered during the first and the fifth week of RT. Patients were evaluated in week 6-8 after treatment completion with pelvic MRI scan and rigid proctoscopy. Pathohistological response after surgery was assessed according to tumor regression grading (TRG) categories by Mandard. Twenty biopsy samples taken at diagnosis were used for proteomic analysis, 9 responders (R, TRG 1-2), and 11 non-responders (NR, TRG 3-5), to achieve the maximum range of different molecular features potentially associated with response. Formalin-fixed paraffin-embedded (FFPE) biopsies were processed, and isolated proteins were digested with trypsin. The resulting peptides were analyzed by liquid chromatography coupled to a Q Exactive HF-X mass spectrometer operated in data independent mode (DIA-MS). Data analysis was performed with DIA-NN and Perseus. Data are available via ProteomeXchange with the identifier PXD040451. Results: The use of DIA-MS allowed the identification and quantification of more than 3,000 proteins per sample in general, a significant increase when compared to the 1,000 proteins previously identified by Data Dependent Acquisition-MS (DDA-MS) in LARC FFPE samples. In total, 4,849 proteins were identified in 20 rectal cancer FFPE samples. Principal Component Analysis (PCA) indicated that responders had a significantly different proteomic profile than non-responders. Statistical analysis of the two groups resulted in the identification of 915 differentially expressed proteins (DEPs) (215 in responders and 700 in non-responders, p<0.05), and 384 with more stringent criteria (p<0.01). Results indicate that some of the leading signaling pathways that correlate with response include the metabolism of RNA, MYC targets, neutrophil degranulation, cellular transport, and response to stimuli. Conclusions: The DIA-MS approach offered unprecedented proteome coverage for FFPE samples. The differentially expressed proteins and biological processes constitute interesting findings that hold the potential for improving LARC patient management.

BackgroundThe standard initial treatment for locally advanced rectal cancer (LARC) is neoadjuvant chemoradiotherapy (nCRT). In order to select patients who would benefit the most from nCRT, there is a strong need for predictive biomarkers. The aim of this study was to evaluate the role of clinical, pathological, radiological, inflammation-related genetic, and hematological parameters in the prediction of response after nCRT.MethodsIn silicoanalysis of published transcriptomics datasets was conducted to identify the best candidate genes, whose expression will be measured using quantitative Real Time PCR (qRT-PCR) in pretreatment formaline-fixed paraffin-embedded (FFPE) samples. In this study, 75 patients with LARC, between June 2020 and January 2022, were prospectively included. Patients were assessed for tumor response in the 8th week after nCRT completion with pelvic MRI scan and rigid proctoscopy. For patients with a clinical complete response (cCR) and initially distant located tumor no immediate surgery was suggested (“watch and wait” approach). The response after surgery was assessed using histopathological tumor regression grading (TRG) categories from postoperative specimens by Mandard. Responders (R) were defined as patients with cCR without operative treatment, and those with TRG 1 and TRG 2 postoperative categories. Non-responders (NR) were patients classified as TRG 3-5.ResultsResponders group comprised 35 patients (46.6%) and NR group included 53.4% of patients. Analysis of published transcriptomics data identified genes that could predict response to treatment and their significance was assessed in our cohort by qRT-PCR. When comparison was made in the subgroup of patients who were operated (TRG1 vs. TRG4), the expression of IDO1 was significantly deregulated (p<0.05). Among hematological parameters between R and NR a significant difference in the response was detected for neutrophil-to-monocyte ratio (NMR), initial basophil, eosinophil and monocyte counts (p<0.01). According to MRI findings, non-responders were more often presented with extramural vascular invasion (p<0.05).ConclusionBased on logistic regression model, factors associated with favorable response to nCRT were found to be tumor morphology as well as hematological parameters which can be easily and routinely derived from initial laboratory results (NMR, eosinophil, basophil and monocyte counts) in a minimally invasive manner. Using various metrics, an aggregated score of the initial eosinophil, basophil, and monocyte counts demonstrated the best predictive performance.

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Maize is one of the most important crop species worldwide, but also extremely susceptible to the effects of increasingly higher temperatures and drought during the summer and its flowering and grain filling stage. Different strategies are being utilized to ensure a satisfying yield potential and quality even in the extremely unfavourable environmental conditions, which are the result of climate change. Some of them are cropping pattern changes and sowing alterations, including earlier sowing. Since this implies the exposure to suboptimal temperatures during early developmental stages, it leads to a demand for developing maize lines tolerant to low temperatures during these stages. This research focuses on the first phase of germination, imbibition. Maize tolerance to low temperatures is a complex trait that includes different mechanisms and strategies that all work together to ensure adaptation and survival, such as cell membrane changes, antioxidative system activation, etc. Additionally, the low temperature response of different maize inbreds varies substantially, so recognizing the different ways in which they respond to low temperatures during imbibition and other earlier stages of development is crucial. To accomplish this, seeds of two maize inbred lines of contrasting susceptibility to low temperatures were selected and exposed to control (20? C) and low temperature (8?C) conditions during the first 24h of imbition and then further analyzed to assess their response. This included germination and tetrazolium assays; ascertaining membrane integrity by evaluating cell leakage and lipid peroxidation; determining the antioxidative capacity by assessing superoxide dismutase (SOD) and catalase (CAT) activity; and expression analysis of four genes included in the low temperature response (gibberellin insensitive dwarf 1 gibberellin receptor, gid1; fatty acid desaturases 2 and 6, fad2 and fad6; plastid-lipid-associated 2 protein, pap2). The results showed that, while there is not a significant difference in their germination rate, they differ in their survival rate, with more seeds of the tolerant genotype surviving the low temperature period. Significant differences between them were found in cell leakage (p<0,01), as well as gid1(p<0,05) and fad6 (p<0,05) gene expression assays. The present research brings light to our understanding of the effect of low temperatures on the first germination stage, - imbibition. It highlights the importance of choosing the right inbreds for earlier sowing and points to certain routes that could be taken for improving and accelerating the breeding process for low temperature tolerance.