Accumulation of Polyphenols and Associated Gene Expression in Hairy Roots of Salvia viridis Exposed to Methyl Jasmonate.

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.

Antioxidant and Anti-Inflammatory Activities of Phenolic Acid-Rich Extract from Hairy Roots of Dracocephalum moldavica.

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-1ß (IL-1ß) production in LPS E. coli stimulated monocytes. The level of pro-inflammatory IL-1ß 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 EC50 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-1ß production (47% inhibition) without affecting the normal growth of L929 fibroblast cells.

Effect of Light Conditions on Polyphenol Production in Transformed Shoot Culture of Salvia bulleyana Diels.

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 rolB and rolC 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.

Role of Phytohormones in Biomass and Polyphenol Accumulation in Salvia bulleyana In Vitro Culture.

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.

The Effect of the Stress-Signalling Mediator Triacontanol on Biochemical and Physiological Modifications in Dracocephalum forrestii Culture.

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.

Evaluation of Patient Experiences Regarding Pharmacist-Administrated Vaccination and Attitude towards Future Additional Pharmacy Services in Poland.

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.

In Vitro Strategy for the Enhancement of the Production of Bioactive Polyphenols in Transformed Roots of Salvia bulleyana.

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.

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.

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.

Pre-Vaccination Stress, Post-Vaccination Adverse Reactions, and Attitudes towards Vaccination after Receiving the COVID-19 Vaccine among Health Care Workers.

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.

The Antioxidant, Cytotoxic and Antimicrobial Potential of Phenolic Acids-Enriched Extract of Elicited Hairy Roots of Salvia bulleyana.

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 IC50 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.

Cytokinin-Based Tissue Cultures for Stable Medicinal Plant Production: Regeneration and Phytochemical Profiling of Salvia bulleyana Shoots.

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.

The effect of different light treatments on morphogenesis, phenolic compound accumulation and antioxidant potential of Dracocephalum forrestii transformed shoots cultured in vitro.

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.

The Protective Function and Modification of Secondary Metabolite Accumulation in Response to Light Stress in Dracocephalum forrestii Shoots.

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.

Barriers Associated with the Uptake Ratio of Seasonal Flu Vaccine and Ways to Improve Influenza Vaccination Coverage among Young Health Care Workers in Poland.

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.

Knowledge, Beliefs and Attitudes towards the Influenza Vaccine among Future Healthcare Workers in Poland.

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.

Cytokinin Signaling and De Novo Shoot Organogenesis.

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.

Phytochemical Profile and Antioxidant Activity of Aerial and Underground Parts of Salvia bulleyana Diels. Plants.

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 (Fe3+) 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.

Transformed Shoots of Dracocephalum forrestii W.W. Smith from Different Bioreactor Systems as a Rich Source of Natural Phenolic Compounds.

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 (O2•- 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.

A Comparison of the Attitudes to Influenza Vaccination Held by Nursing, Midwifery, Pharmacy, and Public Health Students and Their Knowledge of Viral Infections.

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.

Establishment of hairy root cultures of Salvia bulleyana Diels for production of polyphenolic compounds.

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.