35 results on '"Grzegorczyk-Karolak I"'
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2. The Influence of Basal Medium on Polyphenol Accumulation in Shoot Cultures of Clerodendrum trichotomum and Clerodendrum colebrookianum .
- Author
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Gomulski J, Krzemińska M, Jochymek M, Kiss AK, and Grzegorczyk-Karolak I
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- Biomass, Phytochemicals chemistry, Phytochemicals metabolism, Clerodendrum chemistry, Clerodendrum metabolism, Polyphenols metabolism, Culture Media chemistry, Plant Shoots metabolism, Plant Shoots growth & development, Plant Shoots chemistry
- Abstract
Plants of the Clerodendrum genus, known for their rich phytochemical profiles, are used in traditional Chinese, Korean, Japanese, and Indian medicine to treat various ailments, including inflammation, hypertension, diabetes, hyperlipidemia, and cancer. Due to the limited natural availability of these plants, there is a growing interest in utilizing in vitro culture techniques to produce their bioactive compounds sustainably. In this study, the effects are compared of Murashige and Skoog (MS), Woody Plant medium (WP), Gamborg B5 (B5), and Schenk and Hildebrandt (SH) basal media on growth, biomass accumulation, and polyphenolic compound production in shoot cultures of Clerodendrum colebrookianum and Clerodendrum trichotomum . The composition of the culture medium significantly influenced the growth and metabolic profiles of both species. C. trichotomum exhibited the highest proliferation potential on WP and SH media, while C. colebrookianum was similar on WP, SH, and B5 media (multiplication factor of about 20). Dry weight accumulation was highest in C. trichotomum grown on SH medium (0.292 g/culture), while C. colebrookianum achieved a comparable biomass on SH and WP media (0.240 g/culture and 0.228 g/culture, respectively). The chemical analysis showed similar secondary metabolite profiles between the two Clerodendrum species with phenylethanoids such as acteoside being the predominant bioactive compounds in hydromethanolic extracts. WP medium was the most favorable for polyphenol accumulation in C. colebrookianum (64.5 mg/g DW), while the SH medium yielded the highest total polyphenol content in C. trichotomum (36.6 mg/g DW). In this study, the importance is underscored of basal medium selection in optimizing the in vitro production of bioactive polyphenolic compounds in Clerodendrum species, providing a foundation for the sustainable and scalable production of these pharmacologically significant metabolites.
- Published
- 2024
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3. Polyphenol production and gene expression in sage shoot cultures exposed to light-emitting diodes.
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Grzegorczyk-Karolak I, Gawęda-Walerych K, Ejsmont W, Owczarek-Januszkiewicz A, Olszewska M, Grąbkowska R, and Krzemińska M
- Subjects
- Gene Expression Regulation, Plant radiation effects, Artemisia metabolism, Artemisia radiation effects, Artemisia genetics, Rosmarinic Acid, Depsides metabolism, Cinnamates metabolism, Biomass, Polyphenols metabolism, Plant Shoots radiation effects, Plant Shoots metabolism, Plant Shoots growth & development, Light
- Abstract
Sages and their beneficial secondary metabolites have been used in conventional and traditional medicine in many countries, and are extensively studied for their health effects. However, to achieve high production levels, it is crucial to optimize the cultivation conditions. The aim of our study was to determine the optimal light-emitting diode (LED) treatment strategy for promoting plant growth and polyphenol biosynthesis in S. atropatana and S. bulleyana in vitro cultures. Shoots of both species were grown under red, blue, mixed (70 % red and 30 % blue), or white (control) light. The lighting conditions affected not only culture growth and proliferation potential, but also the accumulation of polyphenols and the expression of the genes involved in their biosynthesis (PAL, TAT, RAS). The highest proliferation rates (6.21 for S. atropatana and 4.26 for S. bulleyana) were achieved under white LEDs. In contrast, the highest biomass production was observed under white and mixed red/blue light (both species), although a similar effect was revealed for the blue light treatment for S. bulleyana. The dominant polyphenol in both species was rosmarinic acid: its level was highest in S. atropatana shoots exposed to red light (20.86 mg/g dry weight, DW) and S. bulleyana under white light (19.72 mg/g DW). The effects of the light treatments on gene expression varied between plant species and the analyzed gene; for example, mixed light stimulated RAS expression in S. bulleyana shoots and inhibited it in S. atropatana shoots. Principal component analysis found that gene expression did not always translate directly into rosmarinic acid production. In summary, our findings indicate that optimized lighting conditions have a significant effect on the production of polyphenolic compounds in sage shoot cultures. However, further research is needed to find the relationship between light treatment and plant biosynthetic pathway., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)
- Published
- 2025
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4. Effect of Ag + and Cd 2+ Elicitation on Polyphenol Production in Shoot Culture of Dracocephalum ruyschiana L.
- Author
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Weremczuk-Jeżyna I, Gomulski J, Kiss AK, and Grzegorczyk-Karolak I
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- Silver chemistry, Silver pharmacology, Plant Extracts pharmacology, Plant Extracts chemistry, Flavonoids pharmacology, Flavonoids biosynthesis, Flavonoids metabolism, Antioxidants pharmacology, Antioxidants metabolism, Hydroxybenzoates metabolism, Hydroxybenzoates pharmacology, Chlorophyll metabolism, Polyphenols pharmacology, Polyphenols metabolism, Cadmium metabolism, Plant Shoots drug effects, Plant Shoots metabolism, Plant Shoots growth & development, Lamiaceae metabolism, Lamiaceae chemistry, Lamiaceae growth & development, Lamiaceae drug effects
- Abstract
Abiotic elicitation with heavy metals has demonstrated considerable potential to stimulate the production of industrially important secondary metabolites in plant in vitro cultures. The present study investigates the effect of exogenous silver nitrate and cadmium chloride supplementation on flavonoid and phenolic acid production, as well as other indicators of oxidative stress, in shoot cultures of Dracocephalum ruyschiana L. Owing to the presence of bioactive polyphenolic compounds, this Mongolian medicinal plant is traditionally used as an anti-inflammatory, antibacterial and antipyretic agent. The shoots were cultured for three weeks, and then, cadmium (Cd
2+ ) and silver (Ag+ ) ions (50 or 100 µM) were added to the medium. The maximum proliferation rate was observed in the presence of 100 µM Ag+ (almost 5), the highest chlorophyll content in the presence of 100 µM Cd2+ (0.6 mg/g FW) and the highest biomass was observed with both these treatments (73.4-75.7 g FW and 7.53-7.72 g DW). UPLC-PDA-ESI-MS analysis revealed four phenolic acids and five flavonoid derivatives in the hydromethanolic extract of D. ruyschiana shoots. All treatments stimulated the production of rosmarinic acid (RA), which was the dominant compound in the analyzed culture; the highest level of RA, i.e., about three times higher than the control, was noted in shoots exposed to 50 µM Cd2+ (14.72 mg/g DW), whereas the level of most flavonoids in the culture increased most significantly when exposed to Cd2+ at a concentration of 100 µM. Moreover, the shoots grown in the presence of 100 µM Cd2+ exhibited significantly higher antioxidant potential in comparison to the control. Our findings indicate that heavy metals are able to stimulate phenolic compound biosynthesis in Dracocephalum shoots without any negative impact on their growth. These results could be of significant importance for the medical, nutraceutical and agronomic industries.- Published
- 2024
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5. Clerodendrum trichotomum Thunberg-An Ornamental Shrub with Medical Properties.
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Gomulski J and Grzegorczyk-Karolak I
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- Humans, Phytochemicals chemistry, Phytochemicals pharmacology, Animals, Anti-Inflammatory Agents pharmacology, Anti-Inflammatory Agents chemistry, Clerodendrum chemistry, Plant Extracts chemistry, Plant Extracts pharmacology
- Abstract
Harlequin glory bower ( Clerodendrum trichotomum ) is a shrub or small tree belonging to the Lamiaceae family, native to Japan, Korea, and eastern China. It has esthetic value and in Europe, it is cultivated as an ornamental plant. Its sweet-smelling flowers have a white or pink crown. The calyx turns from green to pink-purple over time, providing an especially decorative touch around surrounding the ripe deep-blue fruits that persist until winter. In the areas of its natural occurrence, the leaves and young shoots of C. trichotomum , and sometimes the roots, flowers and fruits, are used in folk medicine due to its anti-inflammatory, analgesic, anticancer, sedative, and hypotensive effects. Products based on Harlequin glory are also used in the treatment of rheumatoid arthritis, joint pain, skin inflammation, or asthma. Preliminary research on the composition of raw material suggests that its health-promoting effect is associated with the presence of numerous secondary metabolites, including phenylpropanoids, flavonoids, lignans, terpenoids, steroids, alkaloids, and anthraquinones. This work reviews the current state of knowledge about the phytochemistry and in vitro and in vivo pharmacological properties of the extracts and main active components isolated from C. trichotomum . It also indicates that before it can be used in modern medicine, further research is necessary regarding the safety and efficacy of the raw material, its mechanisms of action, and dosage.
- Published
- 2024
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6. Improvement of Bioactive Polyphenol Accumulation in Callus of Salvia atropatana Bunge.
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Grzegorczyk-Karolak I, Ejsmont W, Kiss AK, Tabaka P, Starbała W, and Krzemińska M
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- Antioxidants metabolism, Antioxidants chemistry, Chromatography, High Pressure Liquid, Cinnamates metabolism, Cinnamates chemistry, Rosmarinic Acid, Depsides metabolism, Cotyledon metabolism, Cotyledon chemistry, Naphthaleneacetic Acids pharmacology, Naphthaleneacetic Acids chemistry, Naphthaleneacetic Acids metabolism, Plant Growth Regulators pharmacology, Plant Growth Regulators metabolism, Seedlings metabolism, Seedlings growth & development, Seedlings drug effects, Polyphenols metabolism, Salvia metabolism, Salvia chemistry
- Abstract
Callus cultures of the Iranian medicinal plant Salvia atropatana were initiated from three-week-old seedlings on Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA) and various cytokinins. Although all tested hormonal variants of the medium and explant enabled callus induction, the most promising growth was noted for N -(2-chloro-4-pyridyl)- N '-phenylurea (CPPU)-induced calli. Three lines obtained on this medium (cotyledon line-CL, hypocotyl line-HL, and root line-RL) were preselected for further studies. Phenolic compounds in the callus tissues were identified using UPLC-MS (ultra-performance liquid chromatography-mass spectrometry) and quantified with HPLC (high-performance liquid chromatography). All lines exhibited intensive growth and contained twelve phenolic acid derivatives, with rosmarinic acid predominating. The cotyledon-derived callus line displayed the highest growth index values and polyphenol content; this was exposed to different light-emitting diodes (LED) for improving biomass accumulation and secondary metabolite yield. Under LED treatments, all callus lines exhibited enhanced RA and total phenolic content compared to fluorescent light, with the highest levels observed for white (48.5-50.2 mg/g dry weight) and blue (51.4-53.9 mg/g dry weight) LEDs. The selected callus demonstrated strong antioxidant potential in vitro based on the 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) tests. Our findings confirm that the S. atropatana callus system is suitable for enhanced rosmarinic acid production; the selected optimized culture provide high-quality plant-derived products.
- Published
- 2024
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7. Accumulation of Polyphenols and Associated Gene Expression in Hairy Roots of Salvia viridis Exposed to Methyl Jasmonate.
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Grzegorczyk-Karolak I, Krzemińska M, Grąbkowska R, Gomulski J, Żekanowski C, and Gaweda-Walerych K
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- Chromatography, High Pressure Liquid, Gene Expression, Acetates pharmacology, Biotechnology, Cyclopentanes, Oxylipins
- Abstract
Methyl jasmonate (MJA), a signaling molecule in stress pathways, can be used to induce secondary metabolite synthesis in plants. The present study examines its effects on the growth of Salvia viridis hairy roots, and the accumulation of bioactive compounds, and correlates it with the expression of genes involved in the phenylpropanoid pathway. To our knowledge, this study represents the first exploration of elicitation in S. viridis culture and the first comprehensive analysis of MJA's influence on such a wide array of genes within the polyphenol metabolic pathway in the Salvia genus. Plants were treated with 50 and 100 µM MJA, and samples were collected at intervals of one, three, five, and seven days post-elicitation. HPLC analysis revealed that MJA stimulated the accumulation of all tested compounds, with a 30% increase (38.65 mg/g dry weight) in total polyphenol content (TPC) on day five. Quantitative real-time polymerase chain reaction (RT-PCR) analysis demonstrated a significant increase in the expression of the phenylpropanoid pathway genes- TAT (tyrosine aminotransferase), HPPR (4-hydroxyphenylpyruvate reductase), PAL (phenylalanine ammonia-lyase), C4H (cinnamic acid 4-hydroxylase), 4CL (4-coumarate-CoA ligase), and RAS (rosmarinic acid synthase)-following MJA treatment. For the majority of the genes, this increase was observed after the first day of treatment. Importantly, our present results confirm strong correlations of the analyzed gene expression with polyphenol biosynthesis. These findings support the notion that hairy roots provide a promising biotechnological framework for augmenting polyphenol production. Additionally, the combination of elicitor treatment and transgenic technology emerges as a viable strategy to enhance the biosynthesis of these valuable metabolites.
- Published
- 2024
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8. Antioxidant and Anti-Inflammatory Activities of Phenolic Acid-Rich Extract from Hairy Roots of Dracocephalum moldavica .
- Author
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Weremczuk-Jeżyna I, Gonciarz W, and Grzegorczyk-Karolak I
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- Anti-Inflammatory Agents pharmacology, Anti-Inflammatory Agents chemistry, NF-kappa B, Escherichia coli, Lipopolysaccharides pharmacology, Ether, Lamiaceae, Plant Extracts pharmacology, Plant Extracts chemistry, Antioxidants pharmacology, Antioxidants chemistry
- Abstract
This study evaluates the antioxidant properties and anti-inflammatory potential of polyphenolic acid-rich fractions of 80% methanolic extract from the hairy roots of Dracocephalum moldavica . The fractionation of the crude extract yielded the following: a diethyl ether fraction rich in caffeic acid (DM1) (25.85 mg/g DWE), an n-butyl fraction rich in rosmarinic acid (DM3) (43.94 mg/g DWE) and a water residue rich in salvianolic acid B (DM4) (51.46 mg/g DWE). The content of these compounds was determined using high-performance liquid chromatography (HPLC). Their antioxidant activity was evaluated based on DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt) and FRAP assays. The anti-inflammatory activity of the fractions was determined by their effect on nuclear factor kappa-B (NF-κB) activation and interleukin-1 β (IL-1 β ) production in LPS E. coli stimulated monocytes. The level of pro-inflammatory IL-1 β in cells was measured using ELISA. The activation of NF-κB in THP1-Blue™ cells, resulting in the secretion of SEAP (secreted embryonic alkaline phosphatase), was detected spectrophotometrically using Quanti-Blue reagent. Among the tested fractions, the diethyl ether fraction (DM1) showed the highest antioxidant potential, with an EC
50 value of 15.41 µg/mL in the DPPH assay and 11.47 µg/mL in ABTS and a reduction potential of 10.9 mM Fe(II)/g DWE in FRAP. DM1 at a concentration of 10 mg/mL also efficiently reduced LPS-induced SEAP secretion (53% inhibition) and IL-1 β production (47% inhibition) without affecting the normal growth of L929 fibroblast cells.- Published
- 2023
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9. Effect of Light Conditions on Polyphenol Production in Transformed Shoot Culture of Salvia bulleyana Diels.
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Krzemińska M, Hnatuszko-Konka K, Weremczuk-Jeżyna I, Owczarek-Januszkiewicz A, Ejsmont W, Olszewska MA, and Grzegorczyk-Karolak I
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- Depsides metabolism, Cinnamates metabolism, Antioxidants analysis, Plant Roots chemistry, Rosmarinic Acid, Polyphenols analysis, Salvia chemistry
- Abstract
Various strategies have been used to increase the efficiency of secondary metabolite production in Salvia plants. This report is the first to examine the spontaneous development of Salvia bulleyana shoots transformed by Agrobacterium rhizogenes on hairy roots and the influence of light conditions on the phytochemical profile of this shoot culture. The transformed shoots were cultivated on solid MS medium with 0.1 mg/L of IAA (indole-3-acetic acid) and 1 mg/L of m-Top (meta-topolin), and their transgenic characteristic was confirmed by PCR-based detection of the rol B and rol C genes in the target plant genome. This study assessed the phytochemical, morphological, and physiological responses of the shoot culture under stimulation by light-emitting diodes (LEDs) with different wavelengths (white, WL; blue, B; red, RL; and red/blue, ML) and under fluorescent lamps (FL, control). Eleven polyphenols identified as phenolic acids and their derivatives were detected via ultrahigh-performance liquid chromatography with diode-array detection coupled to electrospray ionization tandem mass spectrometry (UPLC-DAD/ESI-MS) in the plant material, and their content was determined using high-performance liquid chromatography (HPLC). Rosmarinic acid was the predominant compound in the analyzed extracts. The mixed red and blue LEDs gave the highest levels of polyphenol and rosmarinic acid accumulation (respectively, 24.3 mg/g of DW and 20.0 mg/g of DW), reaching two times greater concentrations of polyphenols and three times greater rosmarinic acid levels compared to the aerial parts of two-year-old intact plants. Similar to WL, ML also stimulated regeneration ability and biomass accumulation effectively. However, the highest total photosynthetic pigment production (1.13 mg/g of DW for total chlorophyll and 0.231 mg/g of DW for carotenoids) was found in the shoots cultivated under RL followed by BL, while the culture exposed to BL was characterized as having the highest antioxidant enzyme activities.
- Published
- 2023
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10. Role of Phytohormones in Biomass and Polyphenol Accumulation in Salvia bulleyana In Vitro Culture.
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Grzegorczyk-Karolak I, Krzemińska M, Kiss AK, Owczarek-Januszkiewicz A, and Olszewska MA
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- Polyphenols, Biomass, Plant Shoots, China, Plant Growth Regulators, Salvia
- Abstract
Salvia bulleyana is a plant native to the Chinese Yunnan Province. This species has been used in traditional Chinese medicine as a substitute for Danshen (the roots of Salvia miltiorrhiza ). The aim of our study was to establish an effective system for propagating S. bulleyana shoots to obtain large amounts of material rich in bioactive compounds. Phytohormones were used to regulate shoot growth and regeneration potential and influence plant secondary metabolism. The shoot tips were incubated on a Murashige and Skoog agar medium supplemented with 0.1 or 0.5 mg/L IAA (indole-3-acetic acid) and the cytokinins benzylaminopurine (BAP), meta-topoline (M-T), 6-benzylaminopurine riboside (RBAP), N-benzyl-9-(2-tetrahydropyranyl)-adenine (BPA) or kinetin, (K) at concentrations of 0.5, 1 or 2 mg/L. It was observed that the type and concentration of growth regulator significantly influenced the regeneration potential of S. bulleyana shoots. The highest multiplication rate was obtained when 0.1 mg/L IAA and 2 mg/L BPA were used. Under these conditions, 100% of shoot tips formed buds and almost seven buds/shoot per explant were obtained after five weeks. Meanwhile, the highest biomass was found for shoots growing on a medium supplemented with 0.1 mg/L IAA and 1 mg/L M-T: 1.2 g of fresh weight and 0.17 g of dry weight. However, a medium with 0.1 mg/L IAA and 2 mg/L RBAP was most favorable for bioactive phenolic acid content, with a total polyphenol level (37.7 mg/g dw) 4.5 times higher than in shoots grown on medium without growth regulators (8.23 mg/g dw). Finally, optimal conditions were selected by TOPSIS (technique for order of preference by similarity to the ideal solution); the culture of S. bulleyana grown on an MS medium containing 0.1 mg/L IAA and 1 mg/L M-T was found to be the most efficient for polyphenol accumulation and can be used for the production of medicinally relevant compounds., Competing Interests: The authors declare no conflict of interest.
- Published
- 2023
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11. The Effect of the Stress-Signalling Mediator Triacontanol on Biochemical and Physiological Modifications in Dracocephalum forrestii Culture.
- Author
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Weremczuk-Jeżyna I, Hnatuszko-Konka K, Lebelt L, Piotrowska DG, and Grzegorczyk-Karolak I
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- Signal Transduction, Chlorophyll, Antioxidants pharmacology, Lamiaceae, Fatty Alcohols pharmacology, Plant Growth Regulators pharmacology
- Abstract
Triacontanol (TRIA) has been reported to influence signal transduction in the crosstalk triggered by various stress factors. As a signal player, it is also known to affect many physiological processes, including enhancing the biosynthesis of secondary metabolites. Such knowledge can be used to direct or boost the production of bioactive secondary compounds without stress induction. Therefore, the aim of this study is to evaluate the use of TRIA as a factor stimulating the growth and production of bioactive compounds in the shoot culture of Dracocephalum forrestii . TRIA was applied at three concentrations (2.5, 5, and 10 µM), alone or in combination with phytohormones (6-benzylaminopurine and indole-3-acetic acid). After five weeks, growth and physiochemical parameters (chlorophyll content, antioxidant enzyme activity, and phenolic acid level) were determined. The results indicate that TRIA application significantly increased shoot dry weight, chlorophyll content, antioxidant enzyme activities (superoxide dismutase, peroxidase, and catalase), and total polyphenol level; it also influenced the multiplication ratio in combination with growth regulators. The greatest antioxidant enzyme activity was observed for 5 µM TRIA in hormone-free medium, while the most significant secondary metabolite production was obtained for phytohormone-containing medium supplemented with 10 µM TRIA: total phenolic acid content (19.4 mg/g dry weight) was twice that of the control. Hence, the TRIA application appears to be a valuable biotechnology technique for modifying plant metabolite production.
- Published
- 2022
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12. Evaluation of Patient Experiences Regarding Pharmacist-Administrated Vaccination and Attitude towards Future Additional Pharmacy Services in Poland.
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Grzegorczyk-Karolak I, Zglińska-Pietrzak A, Weremczuk-Jeżyna I, and Kałucka S
- Abstract
In order to increase the number of vaccinations performed during the COVID-19 pandemic in Poland, a significant change was introduced in the legislation allowing for the vaccination to also be performed in pharmacies. A cross-sectional survey was conducted among those who chose a pharmacy as a vaccination point during the pandemic COVID-19. The aim of the study was to determine the overall level of patient satisfaction with pharmacist-administered vaccination in pharmacies and to examine patient opinions regarding the further expansion of pharmacy services. A patient survey was conducted immediately after vaccination in the period from January to March 2022. A total of 398 questionnaires were completed. The respondents reported high satisfaction with pharmacist-administered vaccinations (94.5%). In addition, the majority of consumers felt safe during the vaccination procedure (98.5%), and 88.4% declared they would come for other vaccinations at the pharmacy. The two main reasons for choosing this vaccination place were easy access regarding location (94.2%) and proposed vaccination hours (95.2%). The participants reported high levels of satisfaction concerning the pre-vaccination interview (91.2%), information level before immunization (91.5%), injection technique (96.7%), adjusting the premises to the service (95%) and general care in pharmacy after with vaccination (87.7%). The majority of respondents supported the future expansion of pharmacist-administered services, although the support rate ranged from 52% to 83% depending on the type of service offered. The majority of patients supported the continuation of ongoing prescriptions, blood pressure and blood glucose measurement services by pharmacists. Our findings indicate that the involvement of pharmacies in vaccination programs and other public health services met high levels of patient acceptance.
- Published
- 2022
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13. In Vitro Strategy for the Enhancement of the Production of Bioactive Polyphenols in Transformed Roots of Salvia bulleyana .
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Krzemińska M, Owczarek A, Olszewska MA, and Grzegorczyk-Karolak I
- Subjects
- Phenols, Plant Roots, Polyphenols, Sucrose, Vitamins, Salvia
- Abstract
The underground parts of Salvia bulleyana , a rare Chinese plant species, have long been used in traditional Chinese medicine. The Rhizobium rhizogenes -transformed root culture obtained from this plant might be a promising novel source of valuable phenolics, including rosmarinic acid. The present study identifies for the first time, the optimal growth conditions of S. bulleyana hairy roots regarding production efficiency. The comprehensive optimization comprised cultivation in different basal media (B5, SH, MS, and WP) with full- and half-strength macro- and microelements, different vitamin contents (full, half, one-quarter part, and without) and sucrose concentrations (2, 3, 4, 5%), and under different light conditions: in dark, under blue LED (λ = 430 nm), red LED (λ = 670 nm), mixed blue and red LED (30%:70%), and white LED (390-670 nm). Hairy root growth and bioactive compound accumulation were also detailed every five days over the 50-day culture cycle. The optimal conditions were determined using a technique for order preference by similarity to the ideal solution (TOPSIS). The most efficient combination for root growth and polyphenol content was found to be ½SH liquid medium with half vitamin concentration and 3% sucrose when grown in the dark. The biomass yield during the growth cycle was 6.1 g (fresh weight-FW) and 0.92 g (dry weight-DW) on one Erlenmeyer flask: a 14.3-fold increase in FW and 16.1-fold increase in DW in relation to the inoculum. The highest mean total phenolic content was 93.6 mg/g DW including about 70 mg/g DW rosmarinic acid, reached on day 40 of culture; compared to roots of two-year-old plants grown under field conditions, the total phenolic acid content was four times higher and rosmarinic acid eight times higher. The obtained results place the investigated culture among the best hair root cultures for rosmarinic acid production.
- Published
- 2022
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14. A Retrospective Cross-Sectional Study on the Risk of Getting Sick with COVID-19, the Course of the Disease, and the Impact of the National Vaccination Program against SARS-CoV-2 on Vaccination among Health Professionals in Poland.
- Author
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Kałucka S, Kusideł E, and Grzegorczyk-Karolak I
- Subjects
- COVID-19 Vaccines therapeutic use, Cross-Sectional Studies, Health Personnel, Humans, Poland epidemiology, Retrospective Studies, SARS-CoV-2, Vaccination, COVID-19 epidemiology, COVID-19 prevention & control, Students, Medical
- Abstract
Six months after starting the National Vaccination Program against COVID-19, a cross-sectional retrospective study was conducted among 1200 salaried and non-salaried healthcare workers (HCWs) in Poland. Its aim was to assess factors including the risk of exposure to COVID-19, experiences with COVID-19, the trust in different sources of knowledge about the pandemic and SARS-CoV-2 vaccines, and the government campaign on vaccination as predictors of vaccination acceptance. The strongest awareness of a high risk of work-associated infection was demonstrated by doctors (D) (72.6%) and nurses and midwives (N) (64.8%); however, almost half of the medical students (MS) and nursing and midwifery students (NS) did not identify as a risk group. Out of several dozen variables related to sociodemographic characteristics and personal experience of COVID-19, only occupation, previous COVID-19 infection, and high stress seemed to significantly influence vaccination acceptance. Interestingly, only 6.7% of respondents admitted that the government campaign impacted their decision to vaccinate. This result is not surprising considering that the vast majority of respondents (87.8%) learned about vaccinations from sources such as academic lectures (29.9%), health professionals (29.0%), or the internet (28.9%). Those who gained information about vaccination from traditional media (radio, television, and daily press), a popular platform of the government campaign, had a lower propensity to vaccinate (OR = 0.16, p < 0.001). Additionally, almost twice as many considered the information provided in the campaign to be unreliable. Our findings, from this retrospective study, do not confirm that the government campaign was effective for healthcare professionals. Therefore, in this group, other forms of vaccination incentives should be sought. However, the vaccinated respondents were significantly more likely to support compulsory vaccination against COVID-19 among health professionals.
- Published
- 2022
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15. Pre-Vaccination Stress, Post-Vaccination Adverse Reactions, and Attitudes towards Vaccination after Receiving the COVID-19 Vaccine among Health Care Workers.
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Kałucka S, Kusideł E, Głowacka A, Oczoś P, and Grzegorczyk-Karolak I
- Abstract
The vaccines against COVID-19 are the best treatment for limiting the spread of the epidemic, and from an individual point of view, for avoiding getting sick. A cross-sectional retrospective survey was conducted from 15 May to 15 July 2021 among healthcare workers, including doctors, nurses, midwives, and students at the Medical University of Lodz (nursing, midwifery and medical students) in Poland. Data were obtained from 1080 participants. The aim of the study was to evaluate vaccination coverage against COVID-19 among healthcare workers (HCWs) in Poland, and to analyze their attitude towards the available vaccines, stress before taking, and side effects after administrating them, and motivation towards continuing vaccination in the future (if necessary). The survey also estimated the frequency and quality of adverse post-vaccination reactions after two doses of BioNTech/Pfizer and two doses of AstraZeneca vaccines. The present study revealed that the vaccination ratio after 6 months from the start of vaccination against COVID-19 in Poland among HCWs was very high at 91.2%. However, doctors and medical students were more likely to be vaccinated than nurses and midwives, and nursing and midwifery students (94.8%, 98.3% vs. 78.9% and 86.3%, respectively). The main reasons that HCWs reported receiving a vaccination were to protect the health of themselves and their families, while the main reasons for avoiding a vaccination were fear of side effects, doubts about effectiveness, and an expedited clinical trial process of vaccines. Furthermore, more than two-thirds of those vaccinated reported side effects after receiving at least one dose of the vaccine. Most of the side effects were short-term symptoms with only slight and moderate intensification. The univariate and multivariate logistic regressions showed that the type of vaccine used had a significant impact on the occurrence of adverse post-vaccination effects and the severity and duration of vaccination symptoms. In addition, chronic disease and fear of vaccination also had some influence. Despite this, most participants (more often older than younger participants; p < 0.001) were in favor of compulsory vaccination against COVID-19 for HCWs.
- Published
- 2022
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16. The Antioxidant, Cytotoxic and Antimicrobial Potential of Phenolic Acids-Enriched Extract of Elicited Hairy Roots of Salvia bulleyana .
- Author
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Krzemińska M, Owczarek A, Gonciarz W, Chmiela M, Olszewska MA, and Grzegorczyk-Karolak I
- Subjects
- HeLa Cells, Humans, Plant Extracts chemistry, Anti-Infective Agents pharmacology, Antineoplastic Agents, Phytogenic pharmacology, Antioxidants pharmacology, Hydroxybenzoates chemistry, Plant Extracts pharmacology, Plant Roots chemistry, Salvia chemistry
- Abstract
Hairy root cultures are valuable sources of a range of phytochemicals. Among them, Salvia bulleyana root culture is a promising source of polyphenols, especially rosmarinic acid (RA), a phenolic acid depside with pleiotropic activity and a wide application in medicine and cosmetology. The aim of the study was to enhance the culture productivity by finding suitable elicitation protocol and to determine its biological potential in terms of antioxidant, anticancer and antimicrobial properties. The total content of phenols and the levels of particular constituents in root extracts were analyzed using HPLC-PDA. Among four elicitors tested (yeast extract; methyl jasmonate, MJA; trans-anethol; and cadmium chloride), MJA was found to be the most effective. The greatest boost in phenolic production (up to 124.4 mg/g dry weight) was observed after three-day treatment with MJA at 100 µM, with an almost 100% improvement compared to the controls (non-treated root culture). The hydromethanolic extract from the elicited culture exhibited strong antioxidant activity with IC
50 values of 11.1 µg/mL, 6.5 µg/mL and 69.5 µg/mL for DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid)) and superoxide anion radical, respectively. Moreover, in concentrations of 0.5-5 mg/mL the extract inhibited the growth of LoVo, AGS and HeLa cell lines, but was safe for the L929 cells up to the concentration of 5 mg/mL. The extract also exhibited moderate antimicrobial activity. Thus, the results confirmed that elicitation can be a beneficial strategy for increase the phenolic acid biosynthesis in hairy roots of S. bulleyana , and that such a highly productive culture can show significant biological potential.- Published
- 2022
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17. The effect of different light treatments on morphogenesis, phenolic compound accumulation and antioxidant potential of Dracocephalum forrestii transformed shoots cultured in vitro.
- Author
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Weremczuk-Jeżyna I, Kuźma Ł, and Grzegorczyk-Karolak I
- Subjects
- Flavonoids metabolism, Lamiaceae growth & development, Lamiaceae metabolism, Morphogenesis radiation effects, Photosynthesis, Pigments, Biological metabolism, Antioxidants metabolism, Lamiaceae radiation effects, Light, Phenols metabolism, Plant Shoots metabolism
- Abstract
The aim of this study was to evaluate the effects of wavelengths of light emitted from LEDs on cultured in vitro transformed shoots of Dracocephalum forrestii. The shoots were grown on MS agar medium with 0.5 mg/l BPA (N-benzyl-9-(tetrahydropyranyl)-adenine) and 0.2 mg/l IAA (indole-3-acetic acid) under four light environments: blue, red, red/blue (7:3) and white (control). After four weeks of culture, shoot multiplication rate, biomass and morphology were evaluated, as well as bioactive phenolic content, antioxidant capacities and antioxidant enzyme activities. The hydromethanolic extracts from shoots were analyzed using UHPLC method, and antioxidant potential was evaluated using radical scavenging (1,1-diphenyl-2-picrohydrazyl and superoxide anion), and ferric reducing antioxidant power (FRAP), and enzymatic methods, i.e. sodium dismutase (SOD), catalase (CAT) and peroxidase (POD) activity. It was found that the blue and red/blue light had the strongest effect on morphogenesis and shoot propagation; in these conditions, more than five new shoots were obtained per explant. The blue light cultures demonstrated the highest fresh (0.41 g/tube FW) and dry weights (0.045 g/tube DW), the highest levels of polyphenols (99.7 mg/g DW), i.e. almost three times greater than under white light (35.4 mg/g DW), as well as the highest antioxidant potential. Therefore, LED culture appears to be a beneficial strategy for enhancing the production of the medicinal value of transformed D. forrestii shoot culture., (Copyright © 2021 Elsevier B.V. All rights reserved.)
- Published
- 2021
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18. Cytokinin-Based Tissue Cultures for Stable Medicinal Plant Production: Regeneration and Phytochemical Profiling of Salvia bulleyana Shoots.
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Grzegorczyk-Karolak I, Hnatuszko-Konka K, Krzemińska M, Olszewska MA, and Owczarek A
- Subjects
- Benzyl Compounds pharmacology, Culture Media chemistry, Culture Media pharmacology, Humans, Medicine, Chinese Traditional, Plant Growth Regulators genetics, Plant Leaves drug effects, Plant Leaves growth & development, Plant Shoots drug effects, Plant Shoots growth & development, Plants, Medicinal chemistry, Purines pharmacology, Regeneration drug effects, Salvia growth & development, Cytokinins pharmacology, Plants, Medicinal growth & development, Salvia chemistry, Tissue Culture Techniques
- Abstract
Salvia bulleyana is a rare Chinese medicinal plant that due to the presence of polyphenols lowers the risk of some chronic diseases especially those related to the cardiovascular system. The present study examines the organogenic competence of various combinations and concentrations of plant growth regulators to develop an efficient protocol for in vitro regeneration of S. bulleyana via leaf explants, maintaining the high production of active constituents. The purpose of the study was also to assess the possibilities of using a cytokinin-based regeneration to effectively produce therapeutic compounds. The adventitious shoot formation was observed through direct organogenesis on media with purine derivatives (meta-topolin, mT and benzylaminopurine, BAP), and through indirect organogenesis on media with urea derivatives (tidiazuron, TDZ and forchlorfenuron, CPPU). The highest regeneration frequency (95%) with 5.2 shoots per explant was obtained on leaves cultured on Murashige and Skoog (MS) medium containing 0.1 mg/L naphthalene-1-acetic acid (NAA) and 2 mg/L BAP. Following inter simple sequence repeat (ISSR) marker-based profiling, the obtained organogenic shoot lines revealed a similar banding pattern to the mother line, with total variability of 4.2-13.7%, indicating high level of genetic stability. The similar genetic profile of the studied lines translated into similar growth parameters. Moreover, HPLC analysis revealed no qualitative differences in the profile of bioactive metabolites; also, the total polyphenol content was similar for different lines, with the exception of the shoots obtained in the presence of CPPU that produced higher level of bioactive compounds. This is the first report of an effective and rapid in vitro organogenesis protocol for S. bulleyana , which can be efficiently employed for obtaining stable cultures rich in bioactive metabolites.
- Published
- 2021
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19. The Protective Function and Modification of Secondary Metabolite Accumulation in Response to Light Stress in Dracocephalum forrestii Shoots.
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Weremczuk-Jeżyna I, Hnatuszko-Konka K, Lebelt L, and Grzegorczyk-Karolak I
- Subjects
- Lamiaceae metabolism, Light, Plant Cells metabolism, Plant Shoots metabolism, Secondary Metabolism, Stress, Physiological
- Abstract
The aim of this work was to determine the effect of stress conditions caused by different light sources, i.e., blue LED (λ = 430 nm), red LED (λ = 670 nm), blue and red LED (70%:30%) and white LED (430-670 nm) on the growth and morphology of cultivated in vitro Dracocephalum forrestii shoot culture. It also examines the effects on bioactive phenolic compound production and photosynthetic pigment content, as well as on antioxidant enzyme activity (CAT, SOD, POD) and antioxidant properties. The most beneficial proliferation effect was observed under white LEDs (7.1 ± 2.1 shoots per explant). The white and blue lights stimulated the highest fresh weight gain, while red light induced the highest dry weight gain. The total phenolic acid content ranged from 13.824 ± 1.181 to 20.018 ± 801 mg g DW
-1 depending on light conditions. The highest content of rosmarinic acid was found in the control shoots (cultivated under fluorescent lamps), followed by culture grown under red light. All LED treatments, especially red and blue, increased salvianolic acid B content, and blue increased apigenin p -coumarylrhamnoside biosynthesis. The greatest ferric reduction activity was observed in shoots cultivated under red light, followed by blue; this is associated with the presence of the highest total phenol content, especially phenolic acids. Similarly, the highest DPPH radical scavenging potential was observed under red light followed by blue. This study proves that LEDs have emerged as significant support for directed in vitro propagation, taking advantage of specific stress responses on various light spectra. This study also showed how stress induced by different LED light spectra increases in Dracocephalum forrestii the synthesis of pharmacologically-active compounds. Hence, light stress may turn out to be a simpler alternative to metabolic engineering for improving the production of secondary metabolites of therapeutic value.- Published
- 2021
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20. Barriers Associated with the Uptake Ratio of Seasonal Flu Vaccine and Ways to Improve Influenza Vaccination Coverage among Young Health Care Workers in Poland.
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Kałucka S and Grzegorczyk-Karolak I
- Abstract
Despite not being full-time health care workers, annual flu vaccination is nevertheless an important consideration for medical students. This study examined the reasons for refusing flu vaccination among medical students, a group characterized by low vaccination coverage, despite the fact that the flu vaccine is arguably the most effective way of preventing serious flu complications. A cross-sectional survey was performed of 1313 students at the Medical University of Lodz. The findings indicate that the main sites of vaccination were primary care centers, and main source of information about influenza vaccination (about 90% of cases) was the general practitioner (GP). The most common motivations for vaccination were a recommendation by the family doctor or the belief that it was an important factor for protection against influenza. Most students reported various adverse effects after vaccination, usually mild pain at the site of vaccination, malaise, or fever. The main reasons for rejecting influenza vaccination were the apparent low risk of disease, the need for annual vaccination, the need to pay for it, lack of time or opportunity, lack of vaccination promotion, negative attitudes toward the flu vaccine, or the belief that there are other methods of preventing flu. To increase long-term vaccine acceptance and increase the vaccination rate among medical students and qualified health care workers, there is a need to adapt the health system and to initiate ongoing promotion programs at university to raise consciousness, promote vaccinations, and develop clinical skills for immunization.
- Published
- 2021
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21. Knowledge, Beliefs and Attitudes towards the Influenza Vaccine among Future Healthcare Workers in Poland.
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Kałucka S, Głowacka A, Dziankowska-Zaborszczyk E, and Grzegorczyk-Karolak I
- Subjects
- Attitude of Health Personnel, Female, Health Knowledge, Attitudes, Practice, Health Personnel, Humans, Poland, Pregnancy, Surveys and Questionnaires, Vaccination, Influenza Vaccines, Influenza, Human prevention & control
- Abstract
The flu vaccine is the best treatment for avoiding the flu and its complications. The aim of the study was to evaluate the knowledge of the flu vaccine and attitude towards the influenza vaccine among medical students in four majors of study (Nursing, Midwifery, Pharmacy, and Public health) in all years of study. A total number of 1137 subjects took part in the study. Most of the vaccinated students assessed the flu vaccine positively (78.5%, 73.7%, 60.7%, and 65.1%, according to their respective majors) and reported that they did not get the flu during the period of vaccination (90.4%, 92.1%, 87.4%, and 97.7%, respectively). Therefore, 65% of the students of Pharmacy, 78% of Midwifery, and 83% of Nursing who were vaccinated once in the last three years recommended the influenza vaccination, and 100% of all students received a regular vaccination every year. The univariate and multivariate logistic regressions showed that a maximum of four factors had a significant impact on the students' knowledge of the influenza vaccine. Knowledge about the flu vaccine was the highest among Pharmacy students and lowest among Public health students. Final-year students answered the questions better than the younger ones ( p < 0.05). Their place of residence and flu vaccination status also appeared to influence their answers. Although all students demonstrated good knowledge of the flu vaccine and demonstrated positive attitudes towards the vaccine, their rate of immunization was low. Therefore, health promotion programs are needed to improve immunization coverage among medical students who are future healthcare workers.
- Published
- 2021
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22. Cytokinin Signaling and De Novo Shoot Organogenesis.
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Hnatuszko-Konka K, Gerszberg A, Weremczuk-Jeżyna I, and Grzegorczyk-Karolak I
- Subjects
- Arabidopsis genetics, Arabidopsis growth & development, Gene Expression Regulation, Plant genetics, Plant Growth Regulators genetics, Plant Shoots growth & development, Signal Transduction genetics, Arabidopsis Proteins genetics, Cytokinins genetics, Organogenesis, Plant genetics, Plant Shoots genetics
- Abstract
The ability to restore or replace injured tissues can be undoubtedly named among the most spectacular achievements of plant organisms. One of such regeneration pathways is organogenesis, the formation of individual organs from nonmeristematic tissue sections. The process can be triggered in vitro by incubation on medium supplemented with phytohormones. Cytokinins are a class of phytohormones demonstrating pleiotropic effects and a powerful network of molecular interactions. The present study reviews existing knowledge on the possible sequence of molecular and genetic events behind de novo shoot organogenesis initiated by cytokinins. Overall, the review aims to collect reactions encompassed by cytokinin primary responses, starting from phytohormone perception by the dedicated receptors, to transcriptional reprogramming of cell fate by the last module of multistep-phosphorelays. It also includes a brief reminder of other control mechanisms, such as epigenetic reprogramming.
- Published
- 2021
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23. Phytochemical Profile and Antioxidant Activity of Aerial and Underground Parts of Salvia bulleyana Diels. Plants.
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Grzegorczyk-Karolak I, Krzemińska M, Kiss AK, Olszewska MA, and Owczarek A
- Abstract
Plants have been used for medical purposes since ancient times. However, a detailed analysis of their biological properties and their associated active compounds is needed to justify their therapeutic use in modern medicine. The aim of the study was to identify and quantify the phenolics present in hydromethanolic extracts of the roots and shoots of the Chinese Salvia species, Salvia bulleyana . The qualitative and quantitative analyses were carried out by ultrahigh-performance liquid chromatography with electrospray ionization mass spectrometry detection (UHPLC-PDA-ESI-MS), and high-performance liquid chromatography with photodiode array (HPLC-PDA) detection. The extracts of S. bulleyana were also screened for their antioxidant activity using ferric ion (Fe
3+ ) reducing antioxidant power (FRAP), 1,1-diphenyl-2-picrylhydrazyl (DPPH), diammonium 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) cation (ABTS), superoxide radical anion (O2 • - ), and inhibition of lipid peroxidation assays. The S. bulleyana extracts were found to contain 38 substances, of which 36 were phenols, with a total level of 14.4 mg/g DW (dry weight) in shoots, and 23.1 mg/g DW in roots. Twenty-eight phenols were polyphenolic acids or their derivatives, the most abundant in shoots being rosmarinic acid, and in roots, salvianolic acid K followed by rosmarinic acid. The other major phenolic acids were caffeic acid, caffeoyl-threonic acids, isomers of lithospermic acid, salvianolic acid F, salvianolic acid B, and yunnaneic acid E. In addition to polyphenolic acids, nine flavonoids were detected in the shoot extract. While both extracts showed significant antioxidant activity, the shoot extract, containing both polyphenolic acids and flavonoids, demonstrated a slightly greater antioxidant potential in some of the anti-radical tests than the roots. However, the root extract proved to be slightly more effective in the lipid peroxidation inhibition test. Thus, S. bulleyana was demonstrated as a promising source of antioxidants, and worthy of further more detailed studies.- Published
- 2020
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24. Transformed Shoots of Dracocephalum forrestii W.W. Smith from Different Bioreactor Systems as a Rich Source of Natural Phenolic Compounds.
- Author
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Weremczuk-Jeżyna I, Lisiecki P, Gonciarz W, Kuźma Ł, Szemraj M, Chmiela M, and Grzegorczyk-Karolak I
- Subjects
- Animals, Anti-Bacterial Agents pharmacology, Antioxidants analysis, Bacteria drug effects, Cell Death drug effects, Cell Line, Fungi drug effects, Lamiaceae growth & development, Mice, Microbial Sensitivity Tests, Phytochemicals analysis, Plant Extracts pharmacology, Bioreactors, Lamiaceae chemistry, Phenols analysis, Plant Shoots chemistry, Transformation, Genetic
- Abstract
Transformed shoots of the Tibetan medicinal plant Dracocephalum forrestii were cultured in temporary immersion bioreactors (RITA and Plantform) and in nutrient sprinkle bioreactor (NSB) for 3 weeks in MS (Murashige and Skoog) liquid medium with 0.5 mg/L BPA ( N -benzyl-9-(2-tetrahydropyranyl)-adenine) and 0.2 mg/L IAA (indole-3-acetic acid). The greatest biomass growth index (GI = 52.06 fresh weight (FW) and 55.67 dry weight (DW)) was observed for shoots in the RITA bioreactor, while the highest multiplication rate was found in the NSB (838 shoots per bioreactor). The levels of three phenolic acids and five flavonoid derivatives in the shoot hydromethanolic extract were evaluated using UHPLC (ultra-high performance liquid chromatography). The predominant metabolite was rosmarinic acid (RA)-the highest RA level (18.35 mg/g DW) and total evaluated phenol content (24.15 mg/g DW) were observed in shoots grown in NSB. The NSB culture, i.e., the most productive one, was evaluated for its antioxidant activity on the basis of reduction of ferric ions (ferric reducing antioxidant power, FRAP) and two scavenging radical (O
2 • - and DPPH, 1,1-diphenyl-2-picrylhydrazyl radical) assays; its antibacterial, antifungal, and antiproliative potential against L929 cells was also tested (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) test). The plant material revealed moderate antioxidant and antimicrobial activities and demonstrated high safety in the MTT test-no cytotoxicity at concentrations up to 50 mg/mL was found, and less than a 20% decrease in L929 cell viability was observed at this concentration.- Published
- 2020
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25. A Comparison of the Attitudes to Influenza Vaccination Held by Nursing, Midwifery, Pharmacy, and Public Health Students and Their Knowledge of Viral Infections.
- Author
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Kałucka S, Dziankowska-Zaborszczyk E, Grzegorczyk-Karolak I, and Głowacka A
- Abstract
Influenza is a viral vaccine-preventable disease. The present study aims to explore the attitude to influenza immunization and the knowledge of influenza among students in Poland. A cross-sectional survey was conducted at the end of 2019 and the beginning of 2020 among students of Nursing, Midwifery, Pharmacy, and Public health in all years of study. Data was obtained from 1137 students (90.7% female, 9.3% male), mean age 21.3 ± 1.62 years. The urban students were more likely to be vaccinated against seasonal influenza than rural students (OR: 1.52; 95% CI [1.10-2.10], p = 0.010362). The students of Public health were more likely to be vaccinated against influenza (48.9%, regularly annually 1.1%) than Pharmacy (31%, regularly annually 2.5%), Nursing (30.7%, regularly annually 1.3%) or Midwifery (25.1%, regularly annually 2.4%). First-year and second-year students were vaccinated more often (OR: 2.75; 95% CI [1.99-3.82], p = 0.00000; OR: 1.84; 95% CI [1.32-2.59], p = 0.0004, respectively) than later-year students. All students reported the main reasons for vaccination to be their own protection and parental decision. Concluded, present findings demonstrate a low prevalence of flu vaccination among medical students. Therefore, strategies are needed to increase the uptake of influenza vaccine in students, especially considering the future contact between this group of future health care workers and higher risk groups.
- Published
- 2020
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26. Establishment of hairy root cultures of Salvia bulleyana Diels for production of polyphenolic compounds.
- Author
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Wojciechowska M, Owczarek A, Kiss AK, Grąbkowska R, Olszewska MA, and Grzegorczyk-Karolak I
- Subjects
- Agrobacterium genetics, Cell Culture Techniques, Plant Extracts chemistry, Plant Roots chemistry, Plant Roots genetics, Plants, Genetically Modified, Polyphenols chemistry, Transformation, Genetic, Plant Roots growth & development, Plant Roots metabolism, Polyphenols biosynthesis, Salvia
- Abstract
This study was to obtain stable transformed roots of Salvia bulleyana using A. rhizogenes strain A4 and then evaluate their phytochemical profile and selected the most productive clone. Our results indicated that the type of explant and medium used for bacterium and explant incubation had an influence on the frequency of hairy root formation. The best response was obtained on leaves infected with bacteria cultivated on YMB medium supplemented with acetosyringone. Of the four selected transformed root clones, after five-week cultivation in Woody Plant (WP) medium, the highest growth indexes were demonstrated for line C1: i.e. 13 for fresh and 15 for dry weight (81.4 and 8.2 g/l fresh and dry weight, respectively). The qualitative analysis of hydromethanolic extracts of hairy roots of S. bulleyana using UPLC-PDA-ESI-MS/MS method showed the presence of 10 polyphenolic compounds including predominant rosmarinic acid (RA), its derivatives (hexoside and methyl rosmarinate), caffeic acid, its derivatives and several salvianolic acids: K, E and F. Their production varied among the four root clones studied; the highest RA (39.6 mg/g dry weight) and total polyphenol (48.9 mg/g dry weight) level were found in the roots of C4 clone. These values were significantly higher than those of the roots of plants grown for several years under field conditions. The transformation of the obtained root cultures was confirmed by polymerase chain reaction using aux1, aux2, rolB, rolC and rolD primers., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
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27. The Stimulatory Effect of Purine-Type Cytokinins on Proliferation and Polyphenolic Compound Accumulation in Shoot Culture of Salvia Viridis .
- Author
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Grzegorczyk-Karolak I, Hnatuszko-Konka K, Zarzycka M, and Kuźma Ł
- Subjects
- Cinnamates metabolism, Depsides metabolism, Hydroxybenzoates metabolism, Plant Cells drug effects, Plant Shoots drug effects, Plant Shoots growth & development, Polyphenols biosynthesis, Salvia growth & development, Rosmarinic Acid, Cell Proliferation drug effects, Cytokinins pharmacology, Purines pharmacology, Salvia drug effects
- Abstract
The present study demonstrates hormonal control of Salvia viridis growth and development using four different purine-type cytokinins at different concentrations. The addition of cytokinins significantly increased biomass of cultures, proliferation rate, and, interestingly, secondary metabolite production. The best response in terms of multiplication ratio was recorded on Murashige and Skoog medium supplemented with 0.5 mg/L BPA (N-benzylotetrahydropyranyl adenine), while the greatest biomass accumulation was achieved when supplemented with 1 mg/L m -T (meta-topoline). Quantitative UPLC-DAD analysis of the hydromethanolic extract from S. viridis culture revealed the presence of 12 polyphenols: seven phenolic acids and five phenylethanoids. The highest total content of polyphenolic compounds was found in shoots cultivated on medium with 2 mg/L BPA (18.66 mg/g DW): almost twice that of control shoots. The medium was also the most optimal for the biosynthesis of rosmarinic acid, the predominant phenolic acid. However, the greater phenylethanoid accumulation was stimulated by 1 mg/L m -T: the metabolite content was above three times higher than that found in shoots grown on the control medium (8.03 mg/g DW vs. 2.37 mg/g DW). Hence, it was demonstrated that phytohormones are capable of influencing not only vital physiological processes, but therapeutic potential of plants as well. Therefore, the cytokinin-based sage cultures may be also considered as the alternative sources of bioactive compounds., Competing Interests: The authors declare that there are no conflict of interest.
- Published
- 2020
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28. The effect of purine-type cytokinin on the proliferation and production of phenolic compounds in transformed shoots of Dracocephalum forrestii.
- Author
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Weremczuk-Jeżyna I, Skała E, Kuźma Ł, Kiss AK, and Grzegorczyk-Karolak I
- Subjects
- Agrobacterium genetics, Cinnamates metabolism, Culture Media, Cytokinins chemistry, DNA, Bacterial genetics, Depsides metabolism, Lamiaceae growth & development, Lamiaceae metabolism, Phenols chemistry, Plant Extracts chemistry, Plant Extracts metabolism, Plant Roots genetics, Plant Roots growth & development, Plant Shoots drug effects, Plant Shoots genetics, Plant Shoots growth & development, Plant Shoots metabolism, Plants, Genetically Modified, Transformation, Genetic, Rosmarinic Acid, Cytokinins pharmacology, Lamiaceae drug effects, Lamiaceae genetics, Phenols metabolism, Plant Growth Regulators pharmacology
- Abstract
Dracocephalum forrestii is a perennial, endemic to China plant with a number of pharmaceutical properties. Transformed shoots of the species spontaneously regenerated from hairy roots induced by Agrobacterium rhizogenes. The transgenic nature of the shoots was confirmed by polymerase chain reaction (PCR). The shoot culture was multiplied on Murashige and Skoog (MS) medium with 0.2 mg/l IAA and 0.2, 0.5, 1.0, 2.0 or 5.0 mg/l purine-type cytokinins (mT, BAR, BPA or BAP). The highest multiplication rate (about thirteen shoot or buds per explant) was obtained on MS medium with 0.2 mg/l mT after four weeks of culture. The phenolic compounds present in the hydromethanolic extracts from the D. forrestii transgenic shoots were characterized using UPLC-PDA-ESI-MS. The shoots were found to biosynthesize three phenolic acids and five flavonoid glycosides. UHPLC analysis of the hydromethanolic extracts found the predominant phenolic acid to be rosmarinic acid, with its highest content observed in shoots cultivated with 5.0 mg/l BPA. In contrast, the greatest production of flavonoid derivatives (especially acacetin derivatives) was observed in the medium supplemented with 2 mg/l BPA., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2019
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29. Regeneration of Phaseolus vulgaris from epicotyls and hypocotyls via direct organogenesis.
- Author
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Hnatuszko-Konka K, Kowalczyk T, Gerszberg A, Glińska S, and Grzegorczyk-Karolak I
- Subjects
- Charcoal, Culture Media chemistry, Culture Media pharmacology, Hypocotyl drug effects, Organogenesis, Plant drug effects, Phaseolus drug effects, Plant Growth Regulators pharmacology, Silver Nitrate pharmacology, Hypocotyl growth & development, Organogenesis, Plant physiology, Phaseolus physiology, Tissue Culture Techniques methods
- Abstract
The tissue culture of Phaseolus vulgaris has always been considered difficult. Its regenerative capacity and response to culture conditions are highly genotype-dependent and hamper the application of genetic engineering. The objective of this study was to develop a repeatable technique for organogenic bud induction from selected explants of the common bean. Epicotyls and hypocotyls of six cultivars were investigated to determine the effect of the genotype, and four variants of two basal media (Murashige-Skoog and Gamborg) were tested. The composition of these medium variants was based on the published data suggesting the most universal medium compounds that show the advantage of being applicable to different cultivars. As a result, the common bean epicotyls showed undisputed regeneration superiority over the hypocotyls. Moreover, a well-known observation was confirmed, namely that common bean regeneration is cultivar-specific or at least specific to the cluster of related cultivars. However, efficient regeneration was achieved most often when the epicotyls were incubated on the MS or B5 media amended with AgNO
3 and BAP. Additionally, the positive synergistic influence of activated charcoal and silver nitrate on bud formation was demonstrated. The highest values of the epicotyl in vitro response for the common bean cultivars could be presented as follows: Czerwona (70.00%) > Goldpantera (58.89%) and Ibiza (58.89%) > Plus (55.56%) > Laponia (50.56%) > Złota Saxa (46.11%).- Published
- 2019
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30. Evaluation of antioxidant activity of extracts from the roots and shoots of Scutellaria alpina L. and S. altissima L. in selected blood cells.
- Author
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Grzegorczyk-Karolak I, Kontek B, Kontek R, Wysokińska H, and Olas B
- Subjects
- Antioxidants analysis, Blood Platelets metabolism, Humans, Hydrogen Peroxide pharmacology, Lipid Peroxidation drug effects, Oxidative Stress physiology, Plant Extracts analysis, Plant Roots chemistry, Plant Shoots chemistry, Scutellaria baicalensis, Antioxidants pharmacology, Blood Platelets drug effects, Oxidative Stress drug effects, Plant Extracts pharmacology, Scutellaria chemistry
- Abstract
Background: It is widely known that reactive oxygen species (ROS) can cause oxidative damage in cells and have been linked to the pathogenesis of oxidative diseases, such as atherosclerosis, ischemia, neurodegenerative disease, diabetes, or cancer. Recently, much attention has been focused on preventive strategies for oxidative stress and related diseases. Plants represent a source of bioactive compounds whose antioxidant activity may be useful in protecting against pro-oxidative reactions., Objectives: The study determines the in vitro biological activity of the ethanolic extracts from the shoots and roots of Scutellaria species (S. altissima and S. alpina) in selected blood cells (blood platelets and lymphocytes)., Material and Methods: Platelet activity, both resting and after thrombin stimulation, was used to indicate the ability of the plant extracts to inhibit the production of superoxide anion radicals (O2 •-) and platelet lipid peroxidation. The generation of superoxide anion radicals was measured by cytochrome c reduction. Lipid peroxidation in blood platelets was measured by the level of thiobarbituric acid reactive substances (TBARS). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay was used to determine the protective effect of Scutellaria extracts on lymphocyte cells against oxidative damage induced by hydroxyl radicals., Results: Extracts (5-50 μg/mL) containing phenolic compounds from both Scutellaria species distinctly reduced nonenzymatic lipid peroxidation and arachidonic acid metabolism by blood platelets in vitro. When given at the tested concentration, the extracts reduced the generation of O2 •- in resting blood platelets and platelets activated by thrombin in vitro. All Scutellaria extracts (10 μg/mL) containing phenolic compounds also protected human lymphocytes against oxidative stress induced by hydrogen peroxide (H2O2)., Conclusions: The present study suggests that the natural extracts from S. altissima and S. alpina have antioxidant properties and, therefore, may be beneficial in the prevention of diseases in which blood platelets and lymphocytes are involved, i.e., cancer or inflammatory and infective diseases.
- Published
- 2019
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31. Determination of the Phenolic Profile and Antioxidant Properties of Salvia viridis L. Shoots: A Comparison of Aqueous and Hydroethanolic Extracts.
- Author
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Grzegorczyk-Karolak I and Kiss AK
- Subjects
- Flavonoids isolation & purification, Molecular Structure, Plant Components, Aerial chemistry, Plant Extracts chemistry, Plant Shoots chemistry, Antioxidants analysis, Polyphenols analysis, Salvia chemistry
- Abstract
Salvia viridis L. is an annual herb used in Mediterranean medicine. The purpose of this study was to determine the polyphenol profile of aqueous (decoction and infusion) and hydroethanolic extracts of aerial parts of field-grown S. viridis and to evaluate their antioxidant activity. The polyphenol profiling was performed via UPLC-DAD/ESI-MS. Additionally, the total polyphenol content in extracts tested were determined by UV-Vis spectrophotometry using the Folin-Ciocalteu assay. The antioxidant effect was evaluated by the FRAP, DPPH, ABTS, O₂
•− scavenging and TBARS methods. The hydroethanolic extract gave the highest content of total phenolic compounds, followed by the infusion. The UPLC-DAD/ESI-MS analysis of extracts showed a total of 19 phenolic compounds identified as flavonoids (four compounds), phenylethanoids (eight compounds) and phenolic acids (seven compounds). Rosmarinic acid was the predominant phenolic acid, verbascoside was the predominant phenylethanoid, while apigenin glucuronide or methylluteolin glucuronide, depending on the sample, were the predominant flavonoids in the analyzed extracts. The presence of a high polyphenol level indicated a high antioxidant activity of both the infusion and the hydroalcoholic extract. These results indicate that S. viridis is a rich resource of phenolic compounds and can be used in dietary applications with the potential to reduce oxidative stress.- Published
- 2018
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32. An Untapped Resource in the Spotlight of Medicinal Biotechnology: The Genus Scutellaria.
- Author
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Grzegorczyk-Karolak I, Wiktorek-Smagur A, and Hnatuszko-Konka K
- Subjects
- Humans, Plant Extracts pharmacology, Biotechnology methods, Phytotherapy, Scutellaria chemistry, Scutellaria genetics, Scutellaria growth & development, Scutellaria metabolism
- Abstract
Background: This review is intended to draw the attention of pharmaceutical and biotechnological communities to the untapped potential of the Scutellaria genus. Skullcaps, as they are more widely known, are found in one of the oldest materia medica in the world, that of ancient Chinese pharmacology, and their numerous wide range of medicinal bioactivities have been studied both in vivo and in vitro. For thousands of years, chemical compounds from the Scutellaria species have been safely used as antitumor, antibacterial, antiviral, anti-inflammatory, antioxidant or hepatoprotective factors., Objective: As these effects are well known, reflected in the presence of Scutellaria plants in national pharmacopoeias, it is clear that the plant has yet enormous unexploited potential. The European pharmacological market has turned to the resources of Scutellaria only in the last two decades, and although the construction and clinical processing of a new drug is a long process, the general impression is that very few medical products in pharmacies have been inspired by the phytochemistry of skullcaps., Conclusion: This paper presents the current state of knowledge on the wealth of Scutellaria chemical compounds with treatment applications, its tissue culture and biotechnological achievements, especially in the context of the production of secondary metabolites., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)
- Published
- 2018
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33. Inhibition of Advanced Glycation End-Product Formation and Antioxidant Activity by Extracts and Polyphenols from Scutellaria alpina L. and S. altissima L.
- Author
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Grzegorczyk-Karolak I, Gołąb K, Gburek J, Wysokińska H, and Matkowski A
- Subjects
- Animals, Antioxidants chemistry, Antioxidants therapeutic use, Cattle, Flavonoids chemistry, Flavonoids pharmacology, Lipid Peroxidation drug effects, Plant Extracts pharmacology, Plant Roots chemistry, Polyphenols pharmacology, Scutellaria chemistry, Serum Albumin, Bovine chemistry, Glycation End Products, Advanced antagonists & inhibitors, Hyperglycemia drug therapy, Plant Extracts chemistry, Polyphenols chemistry
- Abstract
Methanolic extracts from the aerial parts and roots of two Scutellaria species, S. alpina and S. altissima, and five polyphenols from these plants demonstrated a significant ability to inhibit the formation of advanced glycation end-products (AGE) in vitro. S. alpina, which is richer in polyphenolic compounds, had strong antiglycation properties. These extracts demonstrated also high activity in the FRAP (ferric-reducing antioxidant power), antiradical (DPPH) and lipid peroxidation inhibition assays. Among the pure compounds, baicalin was the strongest glycation inhibitor (90.4% inhibition at 100 μg/mL), followed by luteolin (85.4%). Two other flavone glycosides had about half of this activity. Verbascoside was similar to the reference drug aminoguanidine (71.2% and 75.9%, respectively). The strong correlation observed between AGE inhibition and total flavonoid content indicated that flavonoids contribute significantly to antiglycation properties. A positive correlation was also observed between antiglycative and antioxidant activities. The studied skullcap species can be considered as a potential source of therapeutic agents for hyperglycemia-related disorders.
- Published
- 2016
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34. ANTIOXIDANT ACTIVITY OF TISSUE CULTURE-RAISED BALLOTA NIGRA L. PLANTS GROWN EX VITRO.
- Author
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Makowczyńska J, Grzegorczyk-KAROLAK I, and Wysokińska H
- Subjects
- Flavonoids analysis, Phenols analysis, Tissue Culture Techniques, Antioxidants pharmacology, Ballota chemistry, Plant Extracts pharmacology
- Abstract
Antioxidant properties and total phenolic and flavonoid contents were evaluated in methanolic extracts of shoots from Ballota nigra plants initiated in vitro (from nodal explants) and in vivo (from seeds). The plants were grown in greenhouse and in the field, and were analyzed at the vegetative and flowering stages. The shoot extract of wild-grown plants of B. nigra was also investigated. The results indicate that antioxidant potential of the B. nigra extracts seems to be due to their scavenging of free radicals (DPPH assay) and metal reducing (FRAP test), while they were less effective at the prevention of linoleic acid peroxidation (LPO test). The extracts from shoots of in vitro derived plants were found to exhibit the greatest antioxidant properties. The extracts were also characterized by the highest content of phenolic compounds and their level was affected by plant developmental stage. The extracts of shoots collected at the flowering period exhibited higher amounts of phenolics and flavonoids than in the extracts of immature plants. A close correlation between the total phenolic content and flavonoid content and antioxidant activity using the DPPH and FRAP assays was obtained. The results of the present study suggest the use in vitro-derived plants of B. nigra instead of using wild plants for pharmaceutical purposes.
- Published
- 2015
35. Studies on the antioxidant properties of extracts from the roots and shoots of two Scutellaria species in human blood plasma.
- Author
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Grzegorczyk-Karolak I, Wysokińska H, and Olas B
- Subjects
- Antioxidants analysis, Blood drug effects, Flavanones analysis, Flavonoids analysis, Glucosides analysis, Glucosides pharmacology, Humans, Hydrogen Peroxide pharmacology, Lipid Peroxidation drug effects, Molecular Structure, Oxidative Stress drug effects, Phenols analysis, Phenols pharmacology, Plant Extracts analysis, Plant Roots chemistry, Plant Shoots chemistry, Antioxidants pharmacology, Plant Extracts pharmacology, Scutellaria chemistry
- Abstract
We determined the in vitro antioxidant activity of methanolic extracts from the shoots and roots of Scutellaria species (S. altissima and S. alpina) against the action of strong oxidants: hydrogen peroxide (H2O2) and H2O2+Fe(2+) (donor of hydroxyl radicals) on plasma proteins and lipids. Lipid peroxidation in human plasma was measured by the level of thiobarbituric acid reactive species (TBARS). Protein oxidation was measured by quantitation of thiol group. We observed that the extracts (5-50 µg ml(-1)) containing phenolic compounds from both Scutellaria species distinctly reduced oxidation of lipids and proteins in human plasma treated with H2O2. These results also indicated that the extracts have a protective effect against oxidative damage to the human plasma lipids and proteins by induced hydroxyl radical. The main components of the plant materials analysed were flavonoids, present as aglycones (luteolin) or glycosides (cynaroside, baicalin, wogonoside). In all of the extracts, the phenylethanoid verbascoside was also found. The properties of the tested plant extracts were also compared with the action of a well characterised commercial antioxidative polyphenolic extract from the berries of Aronia melanocarpa (Aronox(®)). The comparative studies indicated that the analysed plant extracts were comparable to or even more effective in reducing the oxidation processes than the A. melanocarpa extract. The present study suggests that natural extracts from S. altissima and S. alpina have antioxidant activities and, therefore, may be beneficial in the prevention of diseases related to oxidant stress, such as cancer, cardiovascular, and inflammatory diseases.
- Published
- 2015
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