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Artificial intelligence (AI) has been hailed as the fourth industrial revolution and its influence on people's lives is increasing. The research on AI applications in medicine is progressing rapidly. This revolution shows promise for more precise diagnoses, streamlined workflows, increased accessibility to healthcare services and new insights into ever-growing population-wide datasets. While some applications have already found their way into contemporary patient care, we are still in the early days of the AI-era in medicine. Despite the popularity of these new technologies, many practitioners lack an understanding of AI methods, their benefits, and pitfalls. This review aims to provide information about the general concepts of machine learning (ML) with special focus on the applications of such techniques in cardiovascular medicine. It also sets out the current trends in research related to medical applications of AI. Along with new possibilities, new threats arise - acknowledging and understanding them is as important as understanding the ML methodology itself. Therefore, attention is also paid to the current opinions and guidelines regarding the validation and safety of AI-powered tools.

Introduction: Paradoxically, the literature lacks an assessment of the impact of various factors on subsequent coronary interventions in patients with coronary artery disease (CAD)., Aim: To assess the impact of various factors on subsequent percutaneous transluminal coronary angioplasty (PTCA), as well as to characterize the clinical profile of people undergoing repeated diagnostic coronary angiography without significant coronary artery changes., Material and Methods: We investigated retrospective data from 4041 subjects according to the clinical factors which may affect the occurrence of unplanned future PTCA., Results: The strongest risk factors for subsequent PTCA were significant stenosis of left descending artery (OR = 2.17, 95% CI: 1.09-4.32) during baseline coronary angiography, the atherosclerotic burden (number of critically narrowed vessels) (OR for narrowing lesions in 3 epicardial arteries 12.13, 95% CI: 5.40-27.27), and restenosis in a previously implanted stent (OR = 4.34, 95% CI: 1.96-9.62). A strong positive relationship between total mortality and the number of critically narrowed coronary arteries (during baseline hospitalization) was observed. Patients without significant coronary artery stenosis in two diagnostic angiographies (control group) differed from subjects with hemodynamic relevant CAD in: higher creatinine levels, more frequent presence of chronic obstructive pulmonary disease and more frequent symptoms of intermittent claudication., Conclusions: The results of the study are in accord with real clinical practice. The arteriosclerotic burden is a major cause of recurrent PTCA, but an important clinical issue is the qualification for recurrent coronary-angiography in those patients whose previous coronary angiography did not show significant stenosis, because other clinical causes may explain their symptoms., Competing Interests: The authors declare no conflict of interest., (Copyright © 2020 Termedia Sp. z o. o.)

Background: Drosera spatulata is a source of many compounds such as naphthoquinones, phenolic acids, flavonoids, anthocyanins, and naphthalene derivatives. Unfortunately, the information regarding the biological activity and chemical profile of those compounds is still incomplete. Herein, we investigated the biological activity of 3-O-acetylaleuritolic acid (3-O-AAA) in cancer cell lines., Methods: The cell viability of HeLa, HT-29, MCF7, and MCF12A cells was assessed using MTT assay. Proliferation potential was assessed using the clonogenic assay and flow cytometry. Migration modulation was tested using a scratch assay. Protein expression was analyzed by immunoblotting., Results: 3-O-AAA significantly inhibited the growth of all tested tumor cells. The results of the colony formation assay suggested cytostatic properties of the studied compound. The scratch assay showed that 3-O-AAA was an efficient migration inhibitor in a dose-dependent manner. Moreover, it caused modulation of mTOR, beclin1, and Atg5 proteins suggesting a possible role of the compound in autophagy induction., Conclusion: Collectively, these results demonstrated that 3-O-AAA inhibited the proliferation and migration of cancer cell lines as well as contributed to autophagy induction showing some anticancer properties.

In this article we present the correlations observed in the NOESY spectra, which provide direct and unequivocal detection of hydroxyl groups occurring in the chemical structures of three sesquiterpene lactones - a germacranolide (8α-( E ) - 4'-hydroxysenecioyloxy-9α-hydroxyparthenolide) and two guaianolides (cynaropicrin and desacylcynaropicrin 8α-( Z )-(4'-hydroxy-2'-methyl)butenoate). The NOESY spectra and other NMR spectra, which served for the structural determination, are also presented. The data provided in this article are associated with the research article "Phytochemical profiles of the leaves of Stizolophus balsamita and Psephellus sibiricus and their chemotaxonomic implications" Nawrot et al., 2019.

Seven germacranolides - balsamin, izospiciformin, stizolin, 9α-hydroxyparthenolide, 8α-E-(4'-hydroxy)-senecioyloxy-9α-hydroxyparthenolide, stizolicin and 11βH,13-dihydrostizolicin, as well as one undescribed phenol glycoside 3-(3,4-dihydroxyphenyl) propyl senecioate 3-O-β-glucopyranoside were isolated from the leaves of Stizolophus balsamita growing in Iran. Three coumarins, scoparone, scopoletin, umbelliferone and two guaianolides, cynaropicrin and desacylcynaropicrin 8α-(Z)-(4'-hydroxy-2'-methyl)butenoate (= cebellin F) were isolated from the leaves of Psephellus sibiricus. Phytochemical profile of Stizolophus balsamita growing in Iran differs from that of Stizolophus balsamita growing in Kazakhstan. Therefore, the taxonomic separation of Stizolophus balsamita from Iran is suggested. The results also suggest that isolated coumarins may be a chemotaxonomic marker of the Psephellus species. Moreover, the presence of germacranolides in Centaureinae species with stout apical spine ended bracts of flower (S. balsamita) and guaianolides in other species with appendages of the bracts without apical spine (P. sibiricus) suggests a possible connection between the chemical structure of the sesquiterpene lactones and morphology of flowers in the species of the Centaureinae subtribe. In addition, we propose a new 1 H NMR approach for the detection of hydroxyl groups in sesquiterpene lactones., (Copyright © 2018. Published by Elsevier Ltd.)

Inflammation, oxidative stress, myocardial injury biomarkers and clinical parameters (longer AF duration, left atrial enlargement, the metabolic syndrome) are factors commonly related to AF recurrence. This study aims to assess the predictive value of laboratory and clinical parameters responsible for early recurrence of atrial fibrillation (ERAF) following cryoballoon ablation (CBA) using statistical assessment and machine learning algorithms. This study group comprised 118 consecutive patients (mean age, 62.5 ± 7.8 years; women 36%) with paroxysmal (54.1%) and persistent (45.9%) AF who underwent their first pulmonary vein isolation (PVI) performed by CBA (Arctic Front Advance 2nd generation 28 mm). The biomarker concentrations were measured at baseline and after CBA in a 24-h follow-up. ERAF was defined as at least a 30-s episode of arrhythmia registered by a 24 h-Holter monitor within the 3 months following the procedure. 56 clinical, laboratory and procedural variables were collected from each patient. We used two classification algorithms: support vector machines, gradient boosted tree. The synthetic minority over-sampling technique (SMOTE) was used to provide a balanced training data set. Within a period of 3 months 21 patients (17.8%) experienced ERAF. The statistical analysis indicated that the lowered levels of post-ablation TnT (p = 0.043) and CK-MB (p = 0.010) with the TnT elevation (p = 0.044) were the predictors of ERAF following CBA. In addition, diabetes and statin treatment were significantly associated with ERAF after CBA (p < 0.05). The machine learning algorithms confirmed the results obtained in the univariate analysis.

Introduction: Hematological indices including red cell distribution width and neutrophil to lymphocyte ratio are proven to be associated with outcomes of acute coronary syndrome. The usefulness of machine learning techniques in predicting mortality after acute coronary syndrome based on such features has not been studied before., Objective: We aim to create an alternative risk assessment tool, which is based on easily obtainable features, including hematological indices and inflammation markers., Patients and Methods: We obtained the study data from the electronic medical records of 5053 patients hospitalized with acute coronary syndrome during a 5-year period. The time of follow-up ranged from 12 to 72 months. A machine learning classifier was trained to predict death during hospitalization and within 180 and 365 days from admission. Our method was compared with the Global Registry of Acute Coronary Events (GRACE) Score 2.0 on a test dataset., Results: For in-hospital mortality, our model achieved a c -statistic of 0.89 while the GRACE score 2.0 achieved 0.90. For six-month mortality, the results of our model and the GRACE score on the test set were 0.77 and 0.73, respectively. Red cell distribution width (HR 1.23; 95% CL 1.16-1.30; P < 0.001) and neutrophil to lymphocyte ratio (HR 1.08; 95% CL 1.05-1.10; P < 0.001) showed independent association with all-cause mortality in multivariable Cox regression., Conclusions: Hematological markers, such as neutrophil count and red cell distribution width have a strong association with all-cause mortality after acute coronary syndrome. A machine-learned model which uses the abovementioned parameters can provide long-term predictions of accuracy comparable or superior to well-validated risk scores.

Background: Increased systemic and local inflammation play a vital role in the pathophysiology of acute coronary syndrome. This study aimed to assess the usefulness of selected machine learning methods and hematological markers of inflammation in predicting short-term outcomes of acute coronary syndrome (ACS)., Methods: We analyzed the predictive importance of laboratory and clinical features in 6769 hospitalizations of patients with ACS. Two binary classifications were considered: significant coronary lesion (SCL) or lack of SCL, and in-hospital death or survival. SCL was observed in 73% of patients. In-hospital mortality was observed in 1.4% of patients and it was higher in the case of patients with SCL. Ensembles of decision trees and decision rule models were trained to predict these classifications., Results: The best performing model for in-hospital mortality was based on the dominance-based rough set approach and the full set of laboratory as well as clinical features. This model achieved 81 ± 2.4% sensitivity and 81.1 ± 0.5% specificity in the detection of in-hospital mortality. The models trained for SCL performed considerably worse. The best performing model for detecting SCL achieved 56.9 ± 0.2% sensitivity and 66.9 ± 0.2% specificity. Dominance rough set approach classifier operating on the full set of clinical and laboratory features identifies presence or absence of diabetes, systolic and diastolic blood pressure and prothrombin time as having the highest confirmation measures (best predictive value) in the detection of in-hospital mortality. When we used the limited set of variables, neutrophil count, age, systolic and diastolic pressure and heart rate (taken at admission) achieved the high feature importance scores (provided by the gradient boosted trees classifier) as well as the positive confirmation measures (provided by the dominance-based rough set approach classifier)., Conclusions: Machine learned models can rely on the association between the elevated inflammatory markers and the short-term ACS outcomes to provide accurate predictions. Moreover, such models can help assess the usefulness of laboratory and clinical features in predicting the in-hospital mortality of ACS patients.

An increased systemic and local inflammation plays a key role in the pathophysiology of acute coronary syndrome (ACS). This review will discuss the role of hematological indices: white blood cells (WBC), neutrophil to lymphocyte ratio (NLR), red cell distribution width (RDW), and platelet indices, that is, platelet to lymphocyte ratio (PLR), mean platelet volume (MPV), and platelet distribution width (PDW) in the case of ACS. In recent years, a strong interest has been drawn to these indices, given that they may provide independent information on pathophysiology, risk stratification, and optimal management. Their low-cost and consequent wide and easy availability in daily clinical practice have made them very popular in the laboratory testing. Furthermore, many studies have pointed at their effective prognostic value in all-cause mortality, major cardiovascular events, stent thrombosis, arrhythmias, and myocardial perfusion disorders in terms of acute myocardial infarction and unstable angina. The most recent research also emphasizes their significant value in the combined analysis with other markers, such as troponin, or with GRACE, SYNTAX, and TIMI scores, which improve risk stratification and diagnosis in ACS patients.

Autophagy is important in the regulation of survival and death signaling pathways in cancer. PKCϵ revealed high transforming potential and the ability to increase cell migration, invasion, and metastasis. Zapotin (5,6,2',6'-tetramethoxyflavone), a natural flavonoid, showed chemopreventive and anticancer properties. Previously, we reported that downmodulation of induced PKCϵ level by zapotin was associated with decreased migration and increased apoptosis in HeLa cell line containing doxycycline-inducible constitutively active PKCϵ (PKCϵA/E, Ala(159) → Glu). Depending on the genetic and environmental content of cells, autophagy may either precede apoptosis or occur simultaneously. The purpose of this study was to assess the effect of zapotin on autophagy. Increasing concentration of zapotin (from 7.5 µM to 30 µM) caused an inhibition of the formation of autophagosomes and a decline in microtubule-associated protein 1 light chain 3 (LC3) protein levels. The gene expression level of major negative regulator of autophagy was noticeably increased. Moreover, the expression of the pivotal autophagy genes was decreased. These changes were accompanied by alternation in autophagy-related protein levels. In conclusion, our results implied that both the antiautophagic and the proapoptosis effect of zapotin in HeLaPKCϵA/E cells are associated with the protein kinase C epsilon signaling pathway and lead to programmed cell death.

Eryngium planum L. (EP) is as a rare medicinal plant with a lot of potentials as pharmaceutical crops. The aim of our study was to assess the effect of subchronic (28-fold) administration of a 70% ethanol extract of EP roots (200 mg/kg, p.o.) on behavioral and cognitive responses in Wistar rats linked with acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), and beta-secretase (BACE-1) mRNA levels and AChE and BuChE activities in the hippocampus and frontal cortex. On the last day of experiment, 30 min after the last dose of EP or Huperzine A (HU), scopolamine (SC) was given at a dose of 0.5 mg/kg b.w. intraperitoneally. The results of a passive avoidance test showed an improvement in long-term memory produced by the EP extract in both scopolamine-induced rats and control group. EP caused an insignificant inhibition of AChE and BuChE activities in the frontal cortex and the hippocampus. EP decreased mRNA AChE, BuChE, and BACE-1 levels, especially in the cortex. Our results suggest that the EP extract led to the improvement of the long-term memory in rats coupled with total saponin content. The mechanism of EP action is probably complicated, since HPLC-MS analysis showed 64 chemical compounds (phenolics, saponins) in the extract of EP roots.

The Ebola virus disease (EVD), formerly known as a hemorrhagic fever and discovered in 1976, is dangerous, highly infectious disease with very high mortality. There are no licensed therapeutics against EVD, although a range of medicines and therapies are currently being evaluated. During the 2014 Ebola outbreak, an experimental drug named ZMapp was administered on an emergency basis to seven patients of which five were recovered. Currently, since February 2015, ZMapp is tested in clinical trials. ZMapp is a mixture (named a cocktail) of three chimaeric monoclonal antibodies (mAbs) of IgG class, which bind to three different epitopes on Ebola surface glycoprotein (GP). ZMapp was created by systematic selection of antibodies from two other three-component cocktails--MB-003 and ZMab the components of which were produced by rapid transient expression method in tobacco species of Australian origin--Nicotiana benthamiana. The ZMapp antibodies of pharmaceutical grade are manufactured in green-house grown N.benthamiana according to the cGMP (current Good Manufacturing Practice), using RAMP platform (Rapid Antibody Manufacturing Platform) and MagnICON system, which utilizes transient expression by magnifection method using viral vectors delivered to plant tissue by a bacterium--Agrobacterium tumefaciens. The applied glycosylation mutant of N.benthamiana (delta XTFT) synthesizes human-like, biantennary N-glycans, with terminal N-acetylglucoseamine and without typical of plants, immunogenic sugar epitopes-beta1,2-linked xylose and alpha1,3-linked fucose. Due to an absence of fucose on N-glycans attached to the Fc domains, the plant-produced anti-Ebola mAbs elicited significantly stronger antibody-dependent cellular cytotoxicity (ADCC) than the analogous anti-Ebola mAbs with fucosylated (alpha1,6-linked fucose) N-glycans produced in a mammalian CHO cell line--the basic expression system for the industrial production of recombinant therapeutical glycoproteins. As far as a vaccine against Ebola virus disease is considered, so-called Ebola Immunogenic Complex (EIC) consisting of assembled molecules of a humanized IgG mAb--6D8 specific for Ebola GPI with GP1 fused to the C-terminus of the heavy chains, was obtained by transient expression in N. benthamiana.

Tobacco smoke contains thousands of ingredients, including those causing serious respiratory and cardiovascular diseases, and which are carcinogens or cancer promoters. Plants and plant products have antioxidant properties and have a protective role against cancer defined as chemoprevention. This paper presents an overview of published experiments on the protective effect against tobacco smoke or its by compounds and raw materials of natural origin, from plants mainly. These were: N-acetyl-L-cysteine, vitamin C, A and E, beta-carotene, lycopene, andrographolide, farnesol, resveratrol, marigold and tea. These studies were carried out on experimental animals or animal or human cells, which were exposed to cigarette smoke or its extract, or components of tobacco smoke. The studies have shown that the mentioned compounds and raw materials have a protective effect against the harmful effects of tobacco smoke. Mechanisms of action were different--the increase of the level of glutathione and antioxidant enzymes, prevention DNA strand breakage and lipid peroxidation, increased accumulation of the transcription factor Nrf2 that controls the expression of antioxidant genes. Authors frequently suggested that the investigated compounds and the materials may have similar protective effects against tobacco smoke in humans.

The approved therapeutic interferons, which are chiefly indicated for a treatment of hepatitis C or hairy cell leukaemia (IFN-α), relapsingl remitting sclerosis multiplex (IFN-β) and chronic granulomatous disease (IFN-γ), are commercially produced by recombinant DNA technology, mainly in bacteria Escherichia coli (IFN-α, IFN-β1b, IFN-γ), rarely in a mammalian cell line CHO (IFN-βla). A serum half-life time of some non-glycosylated IFN-α products was extended by a chemical attachment of a branched polyethylene glycol (PEG) to give PEGylated IFN-α. The therapy with recombinant interferons proves expensive and hence much hope is concerned with their production in other platforms assumed to be cheaper, like transgenic plants. Currently, tobacco, botanically species Nicotiana tabacum, its cultivars and some related species, especially N. benthamiana, is one of the most important plant expression systems tested for the production of therapeutical polypeptides and proteins (so-called biopharmaceuticals or biologics), especially vaccines, by using either greenhouse or field cultivated plants or cell suspension culture. IFN-α subtypes were expressed in tobacco nuclear genom e (IFN-α2a and 2b), chloroplast genome (IFN-α2b) and by transient expression (IFN-α2b). The IFα-a2b chimera fusions with O-glycosylated protein with O-a-rabinogalactans expressed in tobacco BY-2 cell culture showed increased half-life time similar to that obtained by PEGylation. The production of IFN-α2b (non-glycosydated) in tobacco glasshouse or field cultivation has been also elaborated. One report concerned expression of IFN-β but with low yield. N-glycosylated IFN-γ could be efficiently expressed in tobacco protoplast infected with recombinant brome mosaic virus (BMV) with the yield of 5-10% of total extracted protein. This type interferon (non-glycosylated), when expressed in chloroplast genome, proved unstable and could be obtained with reasonable yield as a fusion with GUS (β-glucuronidase).

Selected fractions of ethanolic extracts obtained from leaves and roots of Eryngium planum (Apiaceae) were evaluated in vitro for amebicidal activity against Acanthamoeba castellanii. This free-living ameba is the cause of Acanthamoeba keratitis, which is a painful, vision-impairing disease of the eyes, and chronic granulomatous amebic encephalitis. Treatment is very difficult and not always effective because of encystation, which makes the amebae highly resistant to anti-amebic drugs. The search for novel natural amebicidal agents is still of current interest. Fractions of E. planum ethanolic extract from basal leaves: flavonoid fraction (Lf), flavonoid-saponin fraction (Lf-s), saponin fraction (Ls) and phenolic acids fraction (La) and from roots: saponin fraction (Rs) and phenolic acids fraction (Ra) were assayed for antiamebic activity. In the presence of the saponin fractions and phenolic acid fractions (ranging from 1-5 mg/mL), the number of the trofozoites of Acanthamoeba castellanii viable strain 309 decreased during the experimental period (0-72 h). On the other hand, the flavonoid fraction from leaves showed a stimulating activity on the amebae. Almost all fractions (except the flavonoid fraction) showed a time- and dose-dependent amebistatic activity on the trophozoites. Of the fractions tested, the phenolic acid fraction from roots at the concentration of 5 mg/L showed the amebicidal activity on the trophozoites.

Tobacco and its use was discovered by Christopher Columbus in parallel with the discovery of America. Soon after, tobacco became a known medicinal plant in Europe. Its harmful effects were gradually discovered, especially those of tobacco smoke, and now it is considered a toxic plant. Tobacco leaf has a monograph in German "Hagers Enzyklopädie derArzneistoffe und Drogen", which describes its old, already not valid, medicinal use and clearly shows the toxic effects. Epidemiological studies indicate about 50% lower incidence of Parkinson's disease in smokers than in non-smokers. In turn, studies of the brains of smokers using positron emission tomography showed significantly decreased level of monoamine oxidase B--an enzyme which degrades dopamine--the neurotransmitter which the significant insufficiency of about 80-85%, is responsible for the symptoms of Parkinson's disease. From the tobacco leaves there were isolated MAO-B inhibitors--naphthoquinone--2,3,6-trimethyl-1,4-naphthoquinone and diterpenoid -trans,trans-farnesol, which occur also in tobacco smoke. In the last decade many papers have appeared on the neuroprotective activity of nicotine, the best known component of tobacco. through the effect of this compound on specific nicotinic cholinergic receptors (nAChRs), which interacts with nigrostriatal dopaminergic system as well as the possibility of using nicotine for the treatment of Parkinson's disease and other neurodegenerative diseases. Moreover, tobacco was also found to contain inhibitors of neuronal nitric oxide synthase (nNOS). Tobacco cannot be considered a medicinal plant, but some compounds occurring in that plant may find therapeutic use.

Zapotin, a tetramethoxyflavone, is a natural compound with a wide spectrum of activities in neoplastic cells. Protein kinase C epsilon (PKCε) has been shown to be oncogenic, with the ability to increase cell migration, invasion and survival of tumor cells. Here we report that zapotin inhibits cell proliferation. In wild-type HeLa cells with basal endogenous expression of PKCε, the IC(50) was found to be 17.9 ± 1.6 μM. In HeLa cells overexpressing doxycycline-inducible constitutively active PKCε (HeLaPKCεA/E), the IC(50) was 7.6 ± 1.3 μM, suggesting that PKCε enhances the anti-proliferative effect of zapotin. Moreover, we found that zapotin selectively activated PKCε in comparison with other PKC family members, but attenuated doxycycline-induced PKCε expression. As a result of zapotin treatment for 6, 12 and 24h, the doxycycline-induced levels of the two differently phosphorylated PKCε forms (87 kDa and 95 kDa) were decreased. Migration assays revealed that increasing concentrations of zapotin (from 3.5 to 15 μM) decreased migration of HeLaPKCεA/E cells. Furthermore, zapotin significantly increased the fraction of apoptotic cells in doxycycline-induced (HeLaPKCεA/E) cells after 24h and decreased the levels of Bcl-2, c-Jun, c-Fos. This was accompanied by a degradation of PARP-1. In summary, activation of PKCε and down-modulation of the induced PKCε level by zapotin were associated with decreased migration and increased apoptosis. These observations are consistent with the previously reported chemopreventive and chemotherapeutic action of zapotin., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Interleukins are cytokines of highly pleiotropic activity and they have high potential for application in the treatment of cancer and autoimmune diseases. Trials of recombinant interleukin production in plants relate almost exclusively to tobacco, where through the transformation of the nuclear genome (agroinfection) monomeric (IL-2, IL-4, IL-13, IL-18), homodimeric (IL-10) and single-chain heterodimeric (IL-12) interleukins have been obtained. The expression of IL-10 as a homodimer in the chloroplast genome could not be reached. Expression of the given interleukin was obtained in the leaves, cell culture and culture of hairy roots of tobacco. Interleukins obtained in tobacco showed similar in vitro biological activity as commercial ILs produced mostly in E. coli. Glycosylated IL-13 obtained in tobacco was much more resistant to proteolytic digestion than commercial non-glycosylated IL-13; therefore in the case of sufficiently large production it could be suitable for oral administration in the treatment of type I diabetes.

One of the concepts of global protection of environment is to reduce greenhouse gas emissions, mainly carbon dioxide, into the atmosphere by replacing fossil fuels by the so-called biofuels, which can be obtained from cultivated plants or any plant waste biomass. Currently applied industrial technologies allow the production of biofuels to receive ethanol, mostly from the reserve carbohydrates of sugar cane and corn as well as biodiesel from oil, mainly from rapeseed or oil palm. Tobacco, which provides a high biomass, can be used to produce biogas, bioethanol and biodiesel. The latter derived from oil from seeds and leaves of tobacco has proved useful for driving cars. Modest oil content in tobacco leaves can be increased by the expression of foreign genes encoding its biosynthesis. Promising future source of biofuels is a waste plant biomass consisting mainly of cell walls, which can be subjected to the degradation to produce sugars suitable for fermentation and the production of bioethanol. A number of enzymes needed for efficient degradation of plant cell walls can be produced using recombinant DNA technology in a variety of plants, particularly in chloroplasts of tobacco.

Along with the depreciation of tobacco as a source of nicotine-containing commercial products, the increase of its appreciation as a potential producer of recombinant therapeutical proteins can be observed. Two species of tobacco--Nicotiana tabacum L. and N. benthamiana are easily grown by well established methods of field or green-house cultivation or cell culture, yield high biomass and soluble protein content, can be easily transformed by several methods and are not food for humans or feed for animals. Expression of foreign proteins, including vaccines, can be achieved in those plants either through stable transformation of nuclear or plastid (chloroplast) genomes or by transient transformation using infection with plant virus or bacteria--Agrobacterium tumefaciens (agroinfiltration). The most advanced mode of agrofiltration termed magnifection, which combines benefits of virus and Agrobacterium and depends on using Agrobacterium with viral pro-vectors, enables high-yield and rapid expression of therapeutical proteins, even in a few days, and can be employed on an industrial scale. Expression of many antigenic proteins, which may serve as antiviral, antibacterial, antiprotozoan and anticancer vaccines, and additionally a few autoantigens designed for the treatment of autoimunogenic diseases, like diabetes, have been achieved in tobacco. To date, a vaccine against Newcastle virus disease in poultry produced by tobacco cell culture has been approved for commercial application and several other vaccines are in advanced stage of development. The possibility of a high-level production of vaccines in tobacco against pandemic influenza or anthrax and plague due to a bioterroristic attack, as well as of individualised anticancer vaccines against non-Hodgkin's lymphoma (NHL) in a much shorter period of time than by traditional methods became realistic and hence caused increased interest in tobacco as a high-efficient producer of vaccines not only of specialistic biotechnology firms but also a big pharmaceutical corporation and a department of defence.

Over one hundred of biopharmaceuticals present on the pharmaceutical market--among them mainly therapeutically peptides and proteins such as hormones, enzymes, interferons, blood factors, monoclonal antibodies, vaccines, growth factors and fusion proteins, are manufactured through recombinant DNA technology in cultures of transgenic bacterial (Escherichia coli), yeast (Saccharomyces cerevisiae) and mammalian cells (CHO, BHK) and rarely insect cells or insects (Bombyx mori). High costs of their manufacturing with simultaneous increased demand for existing and new biopharmaceuticals prompt for the search of cheaper methods of production by using transgenic domestic animals and, in particular, in transgenic crop plants. Among many plant species investigated for ability to produce biopharmaceuticals, such like cereals, legumes, oil plants, vegetables and fruit plants, aquatic plants and mosses, the outstanding features posses two tobacco species--Nicotiana tabacum and Nicotiana benthamiana. Foreign proteins can be expressed in those plants not only in nuclear genome, but also easily in chloroplast genome, through transient transformation and propagation of transgenic plant virus--e.g. tobacco mosaic virus (TMV). Tobacco was shown to be useful to produce various therapeutical proteins, even such complex ones as antibodies including secretory immunoglobulin A (sIgA), with partial humanization of N-glycans. Especially important is possibility of rapid preparation, even within a week, vaccines and monoclonal antibodies for clinical tests or personalized anticancer therapy through transient transformation or infection with transgenic virus. Tobacco with foreign gene may be employed for manufacture in two ways--either in field or green-house cultivation or in the form of in vitro suspension cell culture. Among many biopharmaceuticals expressed in tobacco, several vaccines and antibodies are in advanced stages of development, in clinical evaluations and probably close to application in medicine.

Ten lipophilic flavones were isolated from the leaves of Primula veris from field cultivation - the newly described 3'-hydroxy-4',5'-dimethoxyflavone and 3'-methoxy-4',5'-methylenedioxyflavone, the previously known from chemical synthesis 3',4'-dimethoxyflavone, 2',5'-dimethoxyflavone, and also flavone, 2'-hydroxyflavone, 2'-methoxyflavone, 3'-methoxyflavone, 3',4',5'-trimethoxyflavone and 5,6,2',6'-tetramethoxyflavone (zapotin) which were previously known from plants. The same flavones were found in the leaves of P. veris obtained by in vitro propagation. The structural assignments were derived from (1)H NMR, (13)C NMR, EIMS and UV spectral data and the influence of B-ring oxygen substituents on the C-2, C-3 and H-3 NMR resonances in flavones unsubstituted in the A ring is taken into consideration.

The well-known medicinal plant Plantago lanceolata L. (ribwort plantain) was effectively propagated by direct organogenesis from segments of leaves and roots using MS medium supplemented with IAA (11.42 microM), kinetin (9.29 microM) for multiplication and IAA (5.71 microM) for rooting. The plantlets were successfully hardened (80 %) and transferred to field cultivation (100 %). Two lines of callus tissue, derived from leaves and roots, were obtained on MS medium without NH (4)NO (3) and supplemented with 2,4-D (4.52 microM) and kinetin ( 0.46 microM). From plant materials--leaf rosettes from in vitro, leaves from plants in field cultivation obtained by micropropagation, root-derived callus and leaf-derived callus--sixteen phenylethanoid glucosides representing nine different structures were isolated and identified by spectral methods (1D and 2D NMR) as known for the species: lavandulifolioside ( 1), plantamajoside ( 2,) acteoside ( 3); new for the species: leucosceptoside A ( 4), martynoside ( 5), desrhamnosylisoacteoside ( 6), plantainoside D ( 7), desrhamnosylacteoside ( 8) and - 2-(4-hydroxyphenyl)ethyl beta- D-glucopyranosyl-(1-->3)-4- O- trans- and cis- p-coumaroyl-beta- D-glucopyranoside ( 9)--the latter also being found for the first time in nature and named lancetoside. Only plantamajoside ( 2) and acteoside ( 3) were common to all plant materials, the former was the main constituent of calli (1.19 - 2.84 % of dry weight), while the latter was the main constituent of the leaves (1.78 - 10.43 % of dry weight). Flavonoids were present only in plants of field cultivation.

The callus and, for the first time established, shoot cultures of Drosophyllum lusitanicum Link. (Droseraceae) yielded new naphthalene glucoside-5-hydroxy-4-methoxy-2-naphthalenecarboxylic acid 5-O-beta-glucoside (drosophylloside) and 5-hydroxy-4-methoxy-2-naphthalenecarboxylic acid methyl ester besides other phenolics like naphthalenes-5-hydroxy-4-methoxy-2-naphthalenecarboxylic acid (ancistronaphthoic acid B), hydroplumbagin 4-O-glucoside, naphthoquinones-plumbagin and 3-chloroplumbagin, C-glycosylflavones- vitexin, isovitexin, orientin and isoorientin. The pattern of phenolics found supports affinity of Drosophyllum to the families-Droseraceae, Ancistrocladaceae and Dioncophyllaceae.

The regenerated S. virgaurea and S. graminifolia, obtained previously by micropropagation from shoot tips and the callus tissue of S. virgaurea, were analysed for phenolic compounds. Preliminary used 2D-TLC methods (fingerprinting) showed the presence of phenolic glucoside--leiocarposide (S. virgaurea only), C-glycosyflavones--schaftoside and isoschaftoside (S. graminifolia), flavonoids and caffeoylquinic acids, similar to the ones reported earlier for naturally growing plants with the exception of the callus tissue. The latter contained predominantly caffeoylquinic acids, which were isolated and identified by NMR as 5-O-caffeoylquinic and 3,5-di-O-caffeoylquinic acids. The content of leiocarposide (HPLC) and flavonoids (colorimetry) in S. virgaurea was determined.

A mixture of flavonoid glucuronides, consisting of 7-O-glucuronides of kaempferol and quercetin 3-O-rutinosides, 3-O-gentiobiosides and 3-O-glucosides, was isolated from the perianths of Tulipa gesneriana L. var. 'Paradae'. It showed protective activity against the increased (both chloroform and histamine) skin vascular permeability in rabbits. The protective effect, measured as the reduction in leakage of Evans blue, was 59.8% after peritoneal treatment at a dose of 25 mg/kg, while that of troxerutin was 45.5%.

Ten flavonoid C-glycosyl derivatives: orientin (1), isoorientin (2), vitexin (3), isovitexin (4), isovitexin 7,2"-di-O-glucoside (5), isovitexin 7-O-galactoside-2"-O-glucoside (6), two different 6,8-di-C-hexosylapigenins (7, 8), and two different 6-C-hexosyl-8-C-pentosylapigenins (9, 10) have been either produced from flavonoid fractions from Adonis vernalis L. (1, 2) and Crataegus species (3, 4), or isolated from Stellaria media (L.) Vill. (5-10) to study their antioxidative properties. These were found only for two compounds: orientin (1) and isoorientin (2).

The flavonoid fractions of Solidago virgaurea L.S. gigantea Ait., S. canadensis var. canadensis and S. canadensis var. "scabra" flowers were administrated p.o. to rats and showed diuretic activity. Increase in overnight diuresis reached 57-88%. Decrease of overnight excretion of potassium and sodium also occurred after administration of form examined fractions. The flavonoids from S. virgaurea and S. canadensis var. canadensis caused increased excretion of calcium with urine.

Leaves of Hosta ventricosa yielded eight kaempferol glycosides, of which six: the 3-(2G-glucosylrutinoside)-7-glucoside, 3-sophoroside-7-glucoside, 3-rutinoside-7-glucoside, 3-(2G-glucosylrutinoside), 3-sophoroside, 3-rutinoside were fully characterized and two: the 3-xylosylrutinoside-7-glucoside and 3-xylosylrutinoside were tentatively identified. Investigations included 1H-1H COSY and 1H-1H 2D J NMR analysis.

The synthesis of hemisuccinates of some derivatives of oleanolic acid is reported, their inhibitory response to experimentally induced gastric ulcers in rats is examined. The hemisuccinates 13a (sodium salt of 13), 14 and 17 are more effective inhibitors than carbenoxolon-sodium.

The authors report on the synthesis of 3-acetyloleanol acid-28-amids 10, 12-16, 19, 20, 22 and 24. The amids 16 (30%) and 24 (37%) were of most favourable inhibiting effect on the formation of gastric ulcera following pylorus ligature. The ulcer formation caused by p. o. administration of indometacin (Metindol) was in the main reduced by the amids 15 (60%), 16 (34%), and 24 (46%). With both the methods the effect of carbenoxolon sodium amounts to 40 or 50%. In ulcera induced by indometacin, the 11- and 3-oxooleanol acid derivates 12 and 18 revealed stimulating characteristics.