Insights into Molecular Mechanisms of Anticancer Activity of Juniperus communis Essential Oil in HeLa and HCT 116 Cells.

As cancer remains a significant global health challenge, there is an increasing need for novel therapeutic approaches. We investigated the antitumor potential of Juniperus communis berry essential oil on cervical cancer HeLa and colorectal HCT 116 cells. Cytotoxicity was evaluated through the MTT assay, revealing concentration-dependent reductions in cell viability. A clonogenic assay demonstrated long-term cytotoxic effects. Apoptosis markers were assessed via flow cytometric analysis and showed an induction of the intrinsic pathway in both cell lines, demonstrated by the elevated levels of cleaved caspase-3, Bax/Bcl-2 ratio, JC-10 monomer formation, and cytochrome C migration to the cytosol. The treatment inhibited cell-survival pathways in HCT 116 cells and arrested HeLa cells in the S phase. An extensive molecular docking screening provided insight into the binding affinity and interaction patterns of the essential oil components with NADH ubiquinone oxidoreductase and superoxide dismutase enzymes, further confirming the induction of the intrinsic pathway of apoptosis. The obtained in silico and in vitro results indicated the anticancer potential of J. communis berry essential oil as it interferes with cancer cell molecular mechanisms. Our findings highlight J. communis berry essential oil as a promising natural agent with anticancer potential.

Enhancing the Shelf Life of Sous-Vide Red Deer Meat with Piper nigrum Essential Oil: A Study on Antimicrobial Efficacy against Listeria monocytogenes.

Using sous-vide technology in combination with essential oils offers the potential to extend the preservation of food items while preserving their original quality. This method aligns with the growing consumer demand for safer and healthier food production practices. This study aimed to assess the suitability of minimal processing of game meat and the effectiveness of vacuum packaging in combination with Piper nigrum essential oil (PNEO) treatment to preserve red deer meat samples inoculated with Listeria monocytogenes. Microbial analyses, including total viable count (TVC) for 48 h at 30 °C, coliform bacteria (CB) for 24 h at 37 °C, and L. monocytogenes count for 24 h at 37 °C, were conducted. The cooking temperature of the sous-vide was from 50 to 65 °C and the cooking time from 5 to 20 min. Additionally, the study monitored the representation of microorganism species identified through mass spectrometry. The microbiological quality of red deer meat processed using the sous-vide method was monitored over 14 days of storage at 4 °C. The results indicated that the TVC, CB, and L. monocytogenes counts decreased with the temperature and processing time of the sous-vide method. The lowest counts of individual microorganism groups were observed in samples treated with 1% PNEO. The analysis revealed that PNEO, in combination with the sous-vide method, effectively reduced L. monocytogenes counts and extended the shelf life of red deer meat. Kocuria salsicia, Pseudomonas taetrolens, and Pseudomonas fragi were the most frequently isolated microorganism species during the 14-day period of red deer meat storage prepared using the sous-vide method.

In Vitro Evaluation of Anti-Hemolytic and Cytotoxic Effects of Traditional Mexican Medicinal Plant Extracts on Human Erythrocytes and Cell Cultures.

Plant extracts of fifteen plants of ethnomedicinal use in Mexico were analyzed to provide scientific knowledge of their medicinal properties through the evaluation of different biological activities such as anti-hemolytic, antioxidant, and cytotoxic effects in normal cells. Therefore, methanolic extracts were obtained from each of the plants by the Soxhlet extraction. The hemolytic activity in human erythrocytes was evaluated, as was their potential to protect the erythrocyte membrane against the 2,2'-azobis (2-methylpropionamidine) dihydrochloride (AAPH) and 1,1-diphenyl-2-picryl hydrazyl (DPPH) radicals. Finally, the toxicity of the extracts in normal cell cultures of African green monkey kidney cells (Vero) and peripheral blood mononuclear cells (PBMC) was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction method. Most of the extracts showed low hemolytic activity and high anti-hemolytic activity as well as high selectivity indices (SI) and antioxidant effects. Extracts of H. inuloides, J. dioica, and J. spicigera induced cell proliferation of the Vero cells. K. daigremontiana, A. adstringens, S. mexicanum, J. spicigera, L. tridentata, and M. tenuiflora extracts showed PBMC cell proliferation. In the present study, it was observed that the evaluated extracts did not present hemolytic activity, and some presented low toxicity when Vero and PBMC cell cultures were exposed. In conclusion, traditionally used plants possess beneficial health properties, and it is hoped that this study will serve as a basis for understanding the biological effects of traditionally used plants and may complement future studies.

Research progress on plant stress-associated protein (SAP) family: Master regulators to deal with environmental stresses.

Every year, unfavorable environmental factors significantly affect crop productivity and threaten food security. Plants are sessile; they cannot move to escape unfavorable environmental conditions, and therefore, they activate a variety of defense pathways. Among them are processes regulated by stress-associated proteins (SAPs). SAPs have a specific zinc finger domain (A20) at the N-terminus and either AN1 or C2H2 at the C-terminus. SAP proteins are involved in many biological processes and in response to various abiotic or biotic constraints. Most SAPs play a role in conferring transgenic stress resistance and are stress-inducible. The emerging field of SAPs in abiotic or biotic stress response regulation has attracted the attention of researchers. Although SAPs interact with various proteins to perform their functions, the exact mechanisms of these interactions remain incompletely understood. This review aims to provide a comprehensive understanding of SAPs, covering their diversity, structure, expression, and subcellular localization. SAPs play a pivotal role in enabling crosstalk between abiotic and biotic stress signaling pathways, making them essential for developing stress-tolerant crops without yield penalties. Collectively, understanding the complex regulation of SAPs in stress responses can contribute to enhancing tolerance against various environmental stresses through several techniques such as transgenesis, classical breeding, or gene editing.

Enhancing Deer Sous Vide Meat Shelf Life and Safety with Eugenia caryophyllus Essential Oil against Salmonella enterica.

Modern lifestyles have increased the focus on food stability and human health due to evolving industrial goals and scientific advancements. Pathogenic microorganisms significantly challenge food quality, with Salmonella enterica and other planktonic cells capable of forming biofilms that make them more resistant to broad-spectrum antibiotics. This research examined the chemical composition and antibacterial and antibiofilm properties of the essential oil from Eugenia caryophyllus (ECEO) derived from dried fruits. GC-MS analyses identified eugenol as the dominant component at 82.7%. Additionally, the study aimed to extend the shelf life of sous vide deer meat by applying a plant essential oil and inoculating it with S. enterica for seven days at 4 °C. The essential oil demonstrated strong antibacterial activity against S. enterica. The ECEO showed significant antibiofilm activity, as indicated by the MBIC crystal violet test results. Data from MALDI-TOF MS analysis revealed that the ECEO altered the protein profiles of bacteria on glass and stainless-steel surfaces. Furthermore, the ECEO was found to have a beneficial antibacterial effect on S. enterica. In vacuum-packed sous vide red deer meat samples, the anti-Salmonella activity of the ECEO was slightly higher than that of the control samples. These findings underscore the potential of the ECEO's antibacterial and antibiofilm properties in food preservation and extending the shelf life of meat.

Effect of Essential Oils and Dried Herbs on the Shelf Life of Fresh Goat Lump Cheese.

In recent years, the use of natural preservatives in food products has gained significant attention due to their potential health benefits and effectiveness. A standardized microbiological analysis was conducted on Slovak farm-produced lump goat cheese samples to determine the antibacterial activity of dry herbs and essential oils added to vacuum-packed goat cheese. We employed five dried herbs and five essential oils derived from the same plants. The microbiological quality of 145 fresh and vacuum-packed goat cheese samples was assessed. The number of coliform bacteria, total viable count, lactic acid bacteria, and microscopic filamentous fungi were examined in raw cheese samples stored for 12 days at 4 °C. All cheese samples were vacuum-packed (control samples were packed without vacuum). This study evaluated the potential benefits of using essential oils and dried herbs from thyme (Thymus serpyllum L.), black pepper (Piper nigrum L.), clove (Eugenia caryophyllus Thunb.), mint (Mentha × piperita L.), and basil (Ocimum basilicum L.) as preservatives. The essential oils were obtained from Hanus Ltd., Nitra, Slovakia, and were applied at a concentration of 2%. The dried herbs were obtained from Popradský caj (Poprad, Slovakia) and Mäspoma Ltd. (Zvolen, Slovakia). The results showed that all microorganism groups were significantly reduced in cheese samples following the application of essential oils throughout the entire storage period. During the preservation of cheese samples in polyethylene bags used for vacuum packing food, Lactococcus garvieae, L. lactis, Enterobacter cloacae, and Serratia liquefaciens were the most frequently isolated microbiota. Essential oils and dried herbs demonstrated antimicrobial potential during the storage of vacuum-packed goat cheese.

Characterization of Selected Microalgae Species as Potential Sources of Nutrients and Antioxidants.

Microalgae are exceptional organisms from a nutritional perspective, boasting an array of bioactive compounds that have long justified their incorporation into human diets. In this study, we explored the potential of five microalgae species: Nannochloropsis sp., Tetraselmis chuii, Chaetoceros muelleri, Thalassiosira weissflogii, and Tisochrysis lutea. We conducted comprehensive analyses of their nutritional profiles, encompassing protein content, individual amino acid composition, mineral and trace element levels, fatty acid profiles (including saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs)), polyphenol compositions, and vitamin B content. The antioxidant activity of the ethanolic extracts was evaluated using two methods: ABTS and DPPH radical scavenging assay. The total protein content of the microalgae ranged from 34.09 ± 0.39% to 42.45 ± 0.18%, with the highest concentration observed in T. weissflogii. Essential amino acids such as histidine, threonine, lysine, valine, isoleucine, leucine, phenylalanine, and methionine were present in concentrations ranging from 0.53 ± 0.02 to 12.55 ± 2.21 g/16 g N. Glutamic acid emerged as the most abundant amino acid, with concentrations ranging from 6.73 ± 0.82 to 12.55 ± 2.21 g/16 g N. Among the microalgae species, T. chuii exhibited the highest concentrations of calcium (Ca) and manganese (Mn), while C. muelleri showed prominence in magnesium (Mg), sodium (Na), and iron (Fe). T. weissflogii stood out for its potassium (K) content, and T. lutea contained notable amounts of copper (Cu), zinc (Zn), and lead (Pb). Regarding fatty acid profiles, Nannochloropsis sp. and T. chuii were predominantly composed of SFA, while C. muelleri and T. weissflogii were rich in MUFA. PUFAs dominated the fatty acid profile of T. lutea, which also exhibited the most diverse range of polyphenolic substances. We also analyzed the B vitamin content, with T. lutea displaying the highest concentrations of niacin (B3) and riboflavin (B2). Antioxidant activity was confirmed for all microalgae tested using DPPH and ABTS radical IC50 (mg/mL) converted to Trolox equivalent (TEAC). These findings underscore the substantial potential of the examined microalgae species as sources of biologically valuable substances characterized by rapid growth and relatively undemanding cultivation conditions.

Light-responsive transcription factors VvHYH and VvGATA24 mediate wax terpenoid biosynthesis in Vitis vinifera.

The cuticular wax that covers the surfaces of plants is the first barrier against environmental stresses and increasingly accumulates with light exposure. However, the molecular basis of light-responsive wax biosynthesis remains elusive. In grape (Vitis vinifera), light exposure resulted in higher wax terpenoid content and lower decay and abscission rates than controls kept in darkness. Assay for transposase-accessible chromatin with high-throughput sequencing and RNA-seq data were integrated to draw the chromatin accessibility and cis-elements regulatory map to identify the potential action sites. Terpenoid synthase 12 (VvTPS12) and 3-hydroxy-3-methylglutaryl-CoA reductase 2 (VvHMGR2) were identified as grape wax biosynthesis targets, while VvHYH and VvGATA24 were identified as terpenoid biosynthesis activators, as more abundant wax crystals and higher wax terpenoid content were observed in transiently overexpressed grape berries and Nicotiana benthamiana leaves. The interaction between VvHYH and the open chromatin of VvTPS12 was confirmed qualitatively using a dual luciferase assay and quantitatively using surface plasma resonance, with an equilibrium dissociation constant of 2.81 nm identified via the latter approach. Molecular docking simulation implied the structural nature of this interaction, indicating that 24 amino acid residues of VvHYH, including Arg106A, could bind to the VvTPS12 G-box cis-element. VvGATA24 directly bound to the open chromatin of VvHMGR2, with an equilibrium dissociation constant of 8.59 nm. Twelve amino acid residues of VvGATA24, including Pro218B, interacted with the VvHMGR2 GATA-box cis-element. Our work characterizes the mechanism underlying light-mediated wax terpenoid biosynthesis and provides gene targets for future molecular breeding.

Rosmarinus officinalis L. essential oil enhances salt stress tolerance of durum wheat seedlings through ROS detoxification and stimulation of antioxidant defense.

Salt-induced stress poses a significant barrier to agricultural productivity by impeding crop growth. Presently, environmentalists are dedicated to safeguarding food security by enhancing agricultural yields in challenging environments. Biostimulants play a crucial role in mitigating abiotic stresses in crop production, and among these, plant essential oils (EOs) stand out as organic substances with diverse biological effects on living organisms. Among the natural promoters of plant growth, Rosmarinus officinalis L. essential oil (RoEO) has gained considerable attention. Although the manifold effects of essential oils (EOs) on plant growth have been extensively demonstrated, their impact on salt stress tolerance in durum wheat seedlings remains unexplored. This investigation was undertaken to evaluate the biostimulatory capabilities of RoEO on the durum wheat cultivar "Mahmoudi." The effects of three RoEO concentrations (1, 2.5, and 5 ppm) on seed germination, growth establishment, and the induction of salt resistance under salinity conditions (150 mM NaCl) were tested. At 5 ppm, RoEO enhanced seedlings' tolerance to salinity by improving growth and reducing membrane deterioration and oxidative stress-induced damage. The expression profile analyses of seven stress-related genes (TdNHX1, TdSOS1, TdSOD, TdCAT, TdGA20-ox1, TdNRT2.1, and TdGS) using RT-qPCR showed enhancement of several important genes in durum wheat seedlings treated with 5 ppm RoEO, even under control conditions, which may be related to salt stress tolerance. The results indicate that the application of RoEO suggests a possible alternative strategy to increase salt tolerance in durum wheat seedlings towards better growth quality, thus increasing ROS scavenging and activation of antioxidant defense.

Assessment of Anticancer Properties of Argemone mexicana L. and Berberine: A Comparative Study.

Argemone mexicana L. has been used in traditional Mexican medicine. Among its bioactive constituents, berberine (BER) has garnered attention for its cytotoxic properties against different tumor cell lines. This study investigates the in vitro toxicity against HEP-G2 (human hepatocellular carcinoma) and murine lymphoma (L5178Y-R) cells using the MTT assay of the methanol extract (AmexM), sub-partitions of A. mexicana, and BER. Selectivity indices (SIs) were determined by comparing their cytotoxic effects on VERO (monkey kidney epithelial) and PBMC (human peripheral blood mononuclear) non-tumoral cells. Additionally, the anti-hemolytic effect of these treatments was assessed using the AAPH method. The treatment with the most promising activity against tumor cells and anti-hemolytic efficacy underwent further evaluation for toxicity in Artemia salina and antioxidant activities using DPPH, ABTS, and FRAP assays. BER demonstrated an IC50 = 56.86 µg/mL in HEP-G2 cells and IC50 < 5.0 µg/mL in L5178Y-R cells, with SI values of 15.97 and >5.40 in VERO and PBMC cells, respectively. No significant hemolytic effects were observed, although AmexM and BER exhibited the highest anti-hemolytic activity. BER also demonstrated superior antioxidant efficacy, with lower toxicity in A. salina nauplii compared to the control. Additionally, BER significantly attenuated nitric oxide production. This study highlights the antiproliferative effects of A. mexicana, particularly BER, against HEP-G2 and L5178Y-R tumor cell lines, along with its selectivity towards normal cells. Furthermore, its anti-hemolytic and antioxidant potentials were demonstrated, suggesting that BER is a promising candidate for potent chemotherapeutic agents.

Biological Activity and Phytochemical Characteristics of Star Anise (Illicium verum) Essential Oil and Its Anti-Salmonella Activity on Sous Vide Pumpkin Model.

Illicium verum, commonly known as star anise, represents one of the notable botanical species and is recognized for its rich reservoir of diverse bioactive compounds. Beyond its culinary application as a spice, this plant has been extensively utilized in traditional medicine. Given the contemporary emphasis on incorporating natural resources into food production, particularly essential oils, to enhance sensory attributes and extend shelf life, our study seeks to elucidate the chemical composition and evaluate the antibacterial (in vitro, in situ) and insecticidal properties of Illicium verum essential oil (IVEO). Also, microbiological analyses of pumpkin sous vide treated with IVEO after inoculation of Salmonella enterica were evaluated after 1 and 7 days of study. GC/MS analysis revealed a significantly high amount of (E)-anethole (88.4%) in the investigated EO. The disc diffusion method shows that the antibacterial activity of the IVEO ranged from 5.33 (Streptococcus constellatus) to 10.33 mm (Citrobacter freundii). The lowest minimal inhibition concentration was found against E. coli and the minimum biofilm inhibition concertation was found against S. enterica. In the vapor phase, the best antimicrobial activity was found against E. coli in the pears model and against S. sonei in the beetroot model. The application of the sous vide method in combination with IVEO application decreased the number of microbial counts and eliminated the growth of S. enterica. The most isolated microbiota identified from the sous vide pumpkin were Bacillus amyloliquefaciens, B. cereus, B. licheniformis, and Ralstonia picketii. Modifications to the protein composition of biofilm-forming bacteria S. enterica were suggested by the MALDI TOF MS instigations. The IVEO showed insecticidal potential against Harmonia axyridis. Thanks to the properties of IVEO, our results suggest it can be used in the food industry as a natural supplement to extend the shelf life of foods and as a natural insecticide.

Effect of the Addition of Dandelion (Taraxacum officinale) on the Protein Profile, Antiradical Activity, and Microbiological Status of Raw-Ripening Pork Sausage.

The study evaluated the effect of adding dandelion extract on the characteristics of raw-ripening pork sausages while reducing the nitrite addition from 150 to 80 mg/kg. The sausages were made primarily from pork ham (80%) and pork jowl (20%). The process involved curing, preparing the meat stuffing, forming the links, and then subjecting the sausages to a 21-day ripening period. Physicochemical parameters such as pH, water activity, and oxidation-reduction potential were compared at the beginning of production and after the ripening process. The study also examined the impact of ripening on protein metabolism in pork sausages and compared the protein profiles of different sausage variants. The obtained research results indicate that dandelion-leaf extract (Taraxacum officinale) were rich in phenolic acids, flavonoids, coumarins, and their derivatives (LC-QTOF-MS method). Antiradical activity test against the ABTS+* and DPPH radical, and the TBARS index, demonstrated that addition of dandelion (0.5-1%) significantly improved the oxidative stability of raw-ripening sausages with nitrite content reduction to 80 mg/kg. A microbiological evaluation of the sausages was also carried out to assess the correctness of the ripening process. The total number of viable bacteria, lactic acid bacteria, and coliforms were evaluated and subsequently identified by mass spectrometry.

Phytochemical Characterization and Biological Activities of Essential Oil from Satureja montana L., a Medicinal Plant Grown under the Influence of Fertilization and Planting Dates.

The rising demand for safe plant compounds and herbal products that contribute positively to human health is in line with current market trends. Plants belonging to the Satureja genus, particularly the aromatic medicinal S. montana L. from the Lamiaceae family, are well suited to these trends as they serve as pharmaceutical raw materials. This research aimed to assess the influence of sowing date and fertilization doses, as well as their interaction, on the fresh weight, essential oil content, and composition of S. montana. Experimental cultivation involved varying nitrogen and phosphorus levels. The second cut had the highest fresh weight and oil production compared to the first cut. The highest total plant biomass was achieved with autumn sowing and fertilization at 55 kg N/ha and 37 kg P/ha, whereas Spring sowing exhibited higher essential oil production, with the maximum oil % with 74 kg P/ha and oil yield after applying 55 kg N/ha and 74 kg P/ha. The GC-MS analysis revealed that carvacrol was the predominant compound, with it being recommended to grow S. montana in Spring at doses of 55 kg N/ha and 74 kg P/ha for the superior oil yield. Additionally, S. montana essential oil demonstrated notable biological and antimicrobial activity, positioning it as a potential alternative to chemical food preservatives.

Kaempferol as an Alternative Cryosupplement for Bovine Spermatozoa: Cytoprotective and Membrane-Stabilizing Effects.

Kaempferol (KAE) is a natural flavonoid with powerful reactive oxygen species (ROS) scavenging properties and beneficial effects on ex vivo sperm functionality. In this paper, we studied the ability of KAE to prevent or ameliorate structural, functional or oxidative damage to frozen-thawed bovine spermatozoa. The analysis focused on conventional sperm quality characteristics prior to or following thermoresistance tests, namely the oxidative profile of semen alongside sperm capacitation patterns, and the levels of key proteins involved in capacitation signaling. Semen samples obtained from 30 stud bulls were frozen in the presence of 12.5, 25 or 50 µM KAE and compared to native ejaculates (negative control-CtrlN) as well as semen samples cryopreserved in the absence of KAE (positive control-CtrlC). A significant post-thermoresistance test maintenance of the sperm motility (p < 0.001), membrane (p < 0.001) and acrosome integrity (p < 0.001), mitochondrial activity (p < 0.001) and DNA integrity (p < 0.001) was observed following supplementation with all KAE doses in comparison to CtrlC. Experimental groups supplemented with all KAE doses presented a significantly lower proportion of prematurely capacitated spermatozoa (p < 0.001) when compared with CtrlC. A significant decrease in the levels of the superoxide radical was recorded following administration of 12.5 (p < 0.05) and 25 µM KAE (p < 0.01). At the same time, supplementation with 25 µM KAE in the cryopreservation medium led to a significant stabilization of the activity of Mg2+-ATPase (p < 0.05) and Na+/K+-ATPase (p < 0.0001) in comparison to CtrlC. Western blot analysis revealed that supplementation with 25 µM KAE in the cryopreservation medium prevented the loss of the protein kinase A (PKA) and protein kinase C (PKC), which are intricately involved in the process of sperm activation. In conclusion, we may speculate that KAE is particularly efficient in the protection of sperm metabolism during the cryopreservation process through its ability to promote energy synthesis while quenching excessive ROS and to protect enzymes involved in the process of sperm capacitation and hyperactivation. These properties may provide supplementary protection to spermatozoa undergoing the freeze-thaw process.

Changes in inoculated Salmonella enterica subsp. enterica Serovar Enteritidis and other microbiological qualities of vacuum-packed carrot slices after treatment with aqueous extract of Lobularia maritima.

After harvesting, pathogens can infect fresh vegetables in different ways. Pathogenic bacteria associated with fresh vegetables can cause widespread epidemics associated with foodborne illness. The aim of this study was to assess the microbiological quality of carrot slices after treatment with aqueous extracts of Lobularia maritima (AELm) at different concentrations AELm1 (10 mg/mL), AELm2 (5 mg/mL), AELm3 (2.5 mg/mL) and AELm4 (1.25 mg/mL), and Salmonella enterica subsp. enterica serovar Enteritidis, along with vacuum packaging and storage of carrots for 7 days at 4 °C. On days 1. and 7., total viable counts (TVC), and coliforms bacteria (CB), and Salmonella count were all analysed. Microorganisms that were obtained from carrots were identified using MALDI-TOF MS Biotyper Mass Spectrometry. The total viable, coliform bacteria and Salmonella counts were varied by the group of treatment. Higher counts were found in the control group on both days. The most isolated species of bacteria were Salmonella enterica and Pantoea agglomerans on the 1. day and Klebsiella oxytoca on the 7. day. The current study adds useful information for a better understanding of how Salmonella enterica reacts to the effect of AELm and its potential use as a sustainable washing method to eliminate bacteria from freshly cut carrots.

Citrus limon Essential Oil: Chemical Composition and Selected Biological Properties Focusing on the Antimicrobial (In Vitro, In Situ), Antibiofilm, Insecticidal Activity and Preservative Effect against Salmonella enterica Inoculated in Carrot.

New goals for industry and science have led to increased awareness of food safety and healthier living in the modern era. Here, one of the challenges in food quality assurance is the presence of pathogenic microorganisms. As planktonic cells can form biofilms and go into a sessile state, microorganisms are now more resistant to broad-spectrum antibiotics. Due to their proven antibacterial properties, essential oils represent a potential option to prevent food spoilage in the search for effective natural preservatives. In this study, the chemical profile of Citrus limon essential oil (CLEO) was evaluated. GC-MS analysis revealed that limonene (60.7%), ß-pinene (12.6%), and γ-terpinene (10.3%) are common constituents of CLEO, which prompted further research on antibacterial and antibiofilm properties. Minimum inhibitory concentration (MIC) values showed that CLEO generally exhibits acceptable antibacterial properties. In addition, in situ antimicrobial research revealed that vapour-phase CLEO can arrest the growth of Candida and Y. enterocolitica species on specific food models, indicating the potential of CLEO as a preservative. The antibiofilm properties of CLEO were evaluated by MIC assays, crystal violet assays, and MALDI-TOF MS analysis against S. enterica biofilm. The results of the MIC and crystal violet assays showed that CLEO has strong antibiofilm activity. In addition, the data obtained by MALDI-TOF MS investigation showed that CLEO altered the protein profiles of the bacteria studied on glass and stainless-steel surfaces. Our study also found a positive antimicrobial effect of CLEO against S. enterica. The anti-Salmonella activity of CLEO in vacuum-packed sous vide carrot samples was slightly stronger than in controls. These results highlight the advantages of the antibacterial and antibiofilm properties of CLEO, suggesting potential applications in food preservation.

Novel N-pyrocatechoyl and N-pyrogalloyl hydrazone antioxidants endowed with cytotoxic and antibacterial activity.

Over the years, pharmacological agents bearing antioxidant merits arose as beneficial in the prophylaxis and treatment of various health conditions. Hazardous effects of radical species hyperproduction disrupt normal cell functioning, thus increasing the possibility for the development of various oxidative stress-associated disorders, such as cancer. Contributing to the efforts for efficient antioxidant drug discovery, a thorough in vitro and in silico assessment of antioxidant properties of 14 newly synthesized N-pyrocatechoyl and N-pyrogalloyl hydrazones (N-PYRs) was accomplished. All compounds exhibited excellent antioxidant potency against the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. The extensive in silico analysis revealed multiple favorable features of N-PYRs to inactivate harmful radical species, which supported the obtained in vitro results. Also, in silico experiments provided insights into the preferable antioxidant pathways. Prompted by these findings, the cytotoxicity effects and the influence on the redox status of cancer HCT-116 cells and healthy fibroblasts MRC-5 were evaluated. These investigations exposed four analogs exhibiting both cytotoxicity and selectivity toward cancer cells. Furthermore, the frequently uncovered antimicrobial potency of hydrazone-type hybrids encouraged investigations on G+ and G- bacterial strains, which revealed the antibacterial potency of several N-PYRs. These findings highlighted the N-PYRs as excellent antioxidant agents endowed with cytotoxic and antibacterial features.

Anti-Salmonella Activity of Thymus serpyllum Essential Oil in Sous Vide Cook-Chill Rabbit Meat.

Food is generally prepared and vacuum-sealed in a water bath, then heated to a precise temperature and circulated in a sous vide machine. Due to its affordability and ease of use, this cooking method is becoming increasingly popular in homes and food service businesses. However, suggestions from manufacturers and chefs for long-term, low-temperature sous vide cooking raise questions about food safety in the media. In this study, heat treatment with different times and wild thyme essential oil (EO) in sous vide-processed rabbit longissimus dorsi muscle were found to inactivate Salmonella enterica. The rabbit meat samples were vacuum-packed in control groups, in the second group the rabbit meat samples were injected with S. enterica, and in the third group were meat samples infected with S. enterica with Thymus serpylum EO additive. The vacuum-packed samples were cooked sous vide for the prescribed time at 55, 60, and 65 °C. At 5, 15, 30, and 60 min, the quantities of S. enterica, total bacterial counts, and coliform bacteria were measured in groups of sous vide rabbit meat. Microbiological analyses of rabbit meat samples on days 1 and 7 were evaluated. In this study, total viable counts, coliforms bacteria, and number of Salmonella spp. were identified. After incubation, isolates from different groups of microorganisms were identified by the mass spectrometry technique. For each day measured, the test group exposed to a temperature of 55 °C for 5 min had a greater number of total microbiota. The most isolated microorganisms by MALDI-TOF MS Biotyper from the control and treated groups were Lactococcus garvieae and in the treated groups also S. enterica. Based on our analysis of sous vide rabbit meat samples, we discovered that adding 1% of thyme essential oil to the mixture reduced the amount of Salmonella cells and increased the overall and coliform bacterial counts. The microbiological quality of sous vide rabbit meat that was kept for seven days was positively impacted by the addition of thyme essential oil.

Mixture design of α-pinene, α-terpineol, and 1,8-cineole: A multiobjective response followed by chemometric approaches to optimize the antibacterial effect against various bacteria and antioxidant activity.

α-Pinene, α-terpineol, and 1,8-cineole are compounds naturally present in essential oils, although their amounts vary from oil to oil. Although several studies have reported their antibacterial and antioxidant effects, there are few reports on the synergistic or antagonistic effects of their combinations. The objective of this study was to investigate the combined antibacterial effect of these three compounds. To our knowledge, this is the first report on the prediction of their optimal combination using the mixture design approach. The experimental antibacterial activity of the α-pinene, α-terpineol, and 1,8-cineole mixtures depended on the proportion of each compound in the mixture and the target strain, with minimum inhibitory concentrations (MIC) ranging from 0.31 to 1.85 mg/mL. Using the increased simplex-centroid mixture design, the mixture containing 0.33% of each molecule proved to be the most effective against Bacillus cereus and had the lowest MIC values. In addition, α-pinene, α-terpineol, and 1,8-cineole showed significant antioxidant activity against 2,2-picryl-1-hydrazyl radical (DPPH), with IC50 values of 24.53 ± 0.05, 65.63 ± 0.71, and 63.58 ± 0.01 µg/mL, respectively. Statistical planning and the development of utility profiles of the substance mixtures can predict the optimal composition that will exhibit the highest antibacterial activity against B. cereus as well as antioxidant properties. Furthermore, the synergistic effect of the mixtures can contribute significantly to their successful use as natural preservatives in various applications.

Chemical profile, antioxidant and antimicrobial activity of Pinus heldreichii Christ. Distributed in Bulgaria.

Pinus heldreichii Christ. (Bosnian pine), a Tertiary relict and Balkan sub-endemic, has not been comprehensively studied for its essential oil (EO) profile and bioactivity of its different plant parts. This study aimed to determine the EO yield, composition and antimicrobial activity from different parts of P. heldreichii at three different populations (mountains) in Bulgaria. Furthermore, the study assessed the antioxidant activities of plant tissue, including leaves (needles), twigs wood, male and female cones. The EOs yield from different plant parts ranged from 0.09 % (leaves) to 0.74 % (wood of twigs), with monoterpenes being the predominant class. Limonene, α-pinene, ß-caryophyllene, germacrene D, ß-pinene, and ß-myrcene were detected in the EO extracted from all analyzed trees. However, these compounds were not found in the EO extracted from all plant parts of the same trees. Four chemical groups (chemotypes) were identified for EO from twigs, and three chemotypes were identified for EO from leaves. The chemotypes were based on the percent ratio of the main EO constituents (>5 %). Leaves tissue showed the highest values in terms of polyphenols and flavonoids, as well as higher ABTS radical scavenging activity, while the highest antimicrobial activity against Staphylococcus aureus subsp. aureus was seen in the EOs obtained from twigs. This is the first study to identify several chemotypes based on leaf and twigs EO of P. heldreichii distributed in Bulgarian flora. Furthermore, the EO of twigs tips (TT), male cones (MC), and wood of one-two-year-old twigs (WT) of the same trees were reported for the first time. The total polyphenol, flavonoid content, and radical scavenging activity of tissues of annual twigs wood and biennial twigs wood, leaf tissue, MC tissue, and the twigs tips tissue is also reported for the first time in the accessible literature. These findings highlight the potential of P. heldreichii to provide EOs with varying compositions and bioactivities, making them suitable for nutraceutical, pharmacological, and potentially food additive applications. Furthermore, the identification of chemotype accessions in this study suggests their selection for the development of new forest crop as a source for natural products with desirable composition and bioactivity.