Saponin Fractions from Eryngium planum L. Induce Apoptosis in Ovarian SKOV-3 Cancer Cells.

(1) The cytotoxicity and antioxidant activity of different fractions as well as the pro-apoptotic activity of saponin fractions from Eryngium planum L. in SKOV-3 was investigated. (2) In screening studies, the cytotoxicity of six fractions on SKOV-3 was examined by LDH and SRB assays. The most active fractions-triterpenoid saponins-were selected for further investigation. To determine the mechanism of saponin fractions' cytotoxicity, their ability to induce apoptosis was examined via Annexin V assay. The effect of the saponin fractions on caspase 3 activity was measured using a Caspase 3 Assay Kit. The expression of 84 apoptosis-related genes was investigated in cancer cells exposed to saponin fractions from the roots. The radical scavenging capacity of different fractions was determined via DPPH assay. (3) The pronounced cytotoxic effects in SKOV-3 were demonstrated by saponin fractions from the leaves and roots. Those saponin fractions were chosen for further investigation. The treatment of cancer cell lines with saponins obtained from the roots provoked a significant increase in apoptotic cells. In the SKOV-3 cells, saponins caused upregulation of pro-apoptotic genes and a decrease in anti-apoptotic genes. The activation of caspase 3 was correlated with an increased DFFA expression level in the treated SKOV-3 cells. The most active fractions were phenolic acids from the shoots and roots. (4) To the best of our knowledge, the current study is the first to demonstrate that the barrigenol-type triterpenoid saponin fraction from the roots of E. planum inhibits SKOV-3 cell proliferation and induces apoptosis, which may be regulated by the expression of genes mostly specific to a mitochondria-related pathway.

Cytotoxic Effect of Phenylethanoid Glycosides Isolated from Plantago lanceolata L.

The aim of the study is to investigate whether the bioactive compounds isolated from P. lanceolata inflorescences, namely, phenylethanoid glucosides, acteoside, plantamajoside, and a flavonoid, isorhamnetin-3-O-rutinoside-4'-O-glucoside, possessed cytotoxic activity against the selected cancer cell lines. The potential antitumor effects of two phenylethanoid glycosides and one flavonoid were evaluated via MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay on seven human carcinoma cell lines (MCF-7, MDA-MB-231, Caco-2, HepG2, OVCAR-3, U138-MG, U251-MG) and one nontumorigenic mammary epithelial cell line (MCF-12A). For the first time, acteoside was studied in ovarian cancer cell line OVCAR-3, and plantamajoside in all cell lines except breast adenocarcinoma MDA-MB-281 and hepatocarcinoma HepG2. The phenylethanoid glycosides showed stronger cytotoxic activity than that of the glycoside flavonoid. Acteoside and plantamajoside, at concentrations of 200 and 300 µM, respectively, had a highly toxic effect on the selected two cancer cell lines of breast adenocarcinoma MDA-MB-231 and MCF-7, ovarian cancer cell line OVCAR-3, glioblastoma cell line U138-MG, and hepatocarcinoma cell line HepG2. Both glycosides were significantly less cytotoxic towards nontumorigenic cell line MCF-12A; the effect appeared at a concentration of 400 µM. For the first time, the activity of acteoside and plantamajoside was compared in one parallel investigation. The results are discussed against a broad background of existing knowledge on biological effects, their mechanisms, and structure-activity relationships. Phenylethanoids may be potential compounds with cytotoxic activity against the selected cancer types.

Natural Compounds in Liposomal Nanoformulations of Potential Clinical Application in Glioblastoma.

Glioblastoma (GBM) is the most common malignant neoplasm in adults among all CNS gliomas, with the 5-year survival rate being as low as 5%. Among nanocarriers, liposomal nanoformulations are considered as a promising tool for precise drug delivery. The herein presented study demonstrates the possibility of encapsulating four selected natural compounds (curcumin, bisdemethoxycurcumin, acteoside, and orientin) and their mixtures in cationic liposomal nanoformulation composed of two lipid types (DOTAP:POPC). In order to determine the physicochemical properties of the new drug carriers, specific measurements, including particle size, Zeta Potential, and PDI index, were applied. In addition, NMR and EPR studies were carried out for a more in-depth characterization of nanoparticles. Within biological research, the prepared formulations were evaluated on T98G and U-138 MG glioblastoma cell lines in vitro, as well as on a non-cancerous human lung fibroblast cell line (MRC-5) using the MTT test to determine their potential as anticancer agents. The highest activity was exhibited by liposome-entrapped acteoside towards the T98G cell line with IC50 equal 2.9 ± 0.9 µM after 24 hours of incubation. Noteworthy, curcumin and orientin mixture in liposomal formulation exhibited a synergistic effect against GBM. Moreover, the impact on the expression of apoptosis-associated proteins (p53 and Caspase-3) of acteoside as well as curcumin and orientin mixture, as the most potent agents, was assessed, showing nearly 40% increase as compared to control U-138 MG and T98G cells. It should be emphasized that a new and alternative method of extrusion of the studied liposomes was developed.

Efficacy and Safety of Rituximab for New-Onset Generalized Myasthenia Gravis: The RINOMAX Randomized Clinical Trial.

Importance: Rituximab is a third-line option for refractory generalized myasthenia gravis (MG) based on empirical evidence, but its effect in new-onset disease is unknown. Objective: To investigate the efficacy and safety of rituximab compared with placebo as an add-on to standard of care for MG. Design, Setting, and Participants: This randomized, double-blind, placebo-controlled study took place throughout 48 weeks at 7 regional clinics in Sweden. Key inclusion criteria were age older than 18 years, onset of generalized symptoms within 12 months or less, and a Quantitative Myasthenia Gravis (QMG) score of 6 or more. Patients were screened from October 20, 2016, to March 2, 2020. Key exclusion criteria included pure ocular MG, suspected thymoma, previous thymectomy, and prior noncorticosteroid immunosuppressants or high doses of corticosteroids. Interventions: Participants were randomized 1:1 without stratification to a single intravenous infusion of 500 mg of rituximab or matching placebo. Main Outcomes and Measures: Minimal disease manifestations at 16 weeks defined as a QMG score of 4 or less with prednisolone, 10 mg or less daily, and no rescue treatment. Results: Of 87 potentially eligible patients, 25 were randomized to rituximab (mean [SD] age, 67.4 [13.4] years; 7 [28%] female) and 22 to placebo (mean [SD] age, 58 [18.6] years; 7 [32%] female). Compared with placebo, a greater proportion with rituximab met the primary end point; 71% (17 of 24) in the rituximab group vs 29% (6 of 21) in the placebo group (Fisher exact test P = .007; probability ratio, 2.48 [95% CI, 1.20-5.11]). Secondary end points, comparing changes in Myasthenia Gravis Activities of Daily Living and Myasthenia Gravis Quality of Life at 16 weeks with QMG at 24 weeks did not differ between groups with censoring for rescue treatment (per-protocol analysis) but were in favor of active treatment when rescue treatment was taken into account by worst rank imputation (post hoc analysis). Rescue treatments were also more frequent in the placebo arm (rituximab: 1 [4%]; placebo, 8 [36%]). One patient in the placebo arm had a myocardial infarction with cardiac arrest and 1 patient in the active arm experienced a fatal cardiac event. Conclusions and Relevance: A single dose of 500 mg of rituximab was associated with greater probability of minimal MG manifestations and reduced need of rescue medications compared with placebo. Further studies are needed to address long-term benefit-risk balance with this treatment. Trial Registration: ClinicalTrials.gov Identifier: NCT02950155.

The Evaluation of Phenolic Acids and Flavonoids Content and Antiprotozoal Activity of Eryngium Species Biomass Produced by Biotechnological Methods.

Three species from the Eryngium L. genus-E. campestre, E. maritimum, and E. planum, plants with a rich chemical composition, were selected for phytochemical and biological studies. The applied biotechnological methods allowed to obtain the biomass of these rare or protected species in the form of multiplied shoots (stationary system) and roots cultured in a liquid medium (agitated system). In the extracts from the raw material obtained under in vitro conditions, the content of selected phenolic acids and flavonoids (HPLC-DAD method) as well as the total of polyphenols (Folin-Ciocalteu assay) were quantified. The highest amount of all phenolic compounds was found in extracts from E. planum roots (950.90 ± 33.52 mg/100 g d.w.), and the lowest from E. campestre roots (285.00 ± 10.07 mg/100 g d.w.). The quantitatively dominant compound proved to be rosmarinic acid. The highest amounts were confirmed for E. planum root extract (694.58 mg/100 g d.w.), followed by E. planum (388.95 mg/100 g d.w.) and E. campestre (325.85 mg/100 g d.w.) shoot extracts. The total content of polyphenols was always increased in the biomass from in vitro cultures in comparison to the analogous organs of intact plants of each species. The obtained extracts were assessed for antiprotozoal activity against Acanthamoeba sp. The strength of biological activity of the extracts correlated with the content of phenolic compounds. To our knowledge, this is the first report on the amoebicidal activity of E. campestre, E. maritimum, and E. planum extracts from biomass produced by biotechnological methods.

A new flavonoid, a new phenylethanoid glycoside and related compounds isolated from the inflorescences of Plantago lanceolata L.

For the first time inflorescences of a plant species from the genus Plantago (Pantaginaceae)-Plantago lanceolata L. (Ribwort Plantain), a known medicinal plant, were subjected to studies of phenolic compounds, which resulted in an isolation of two new compounds: a flavonoid-isorhamnetin 3-O-α-L-4C1-arabinopyranosyl-(1→2)-ß-D-4C1-glucopyranoside) (1) and a phenylethanoid glycoside-2-(3,4-dihydroxyphenyl)ethyl O-α-L-arabinofuranosyl-(1→2)-[α-L-1C4-rhamnopyranosyl-(1→3)][E-caffeoyl-1→4]-ß-D-4C1-glucopyranoside (14), along with fourteen known compounds-eight flavonoids (2-9) and six phenylethanoid glycosides (10-13, 15-16). The chemical structures were established by 1 D and 2 D NMR and HRESIMS spectral methods. The known phenylethanoids were the same as reported for leaves or aerial parts of P. lanceolata or other Plantago species. The flavonoids appeared to be only flavonols, mainly isorhamnetin 3-O- and 3,4'-O- glycosides, and thus completely different from flavones, mainly luteolin and apigenin glucuronides, previously reported in the leaves. The possible medicinal and chemotaxonomic relevance of the phenolics found in P. lanceolata inflorescences were taken into consideration.

Patient-Reported Symptom Severity in a Nationwide Myasthenia Gravis Cohort: Cross-sectional Analysis of the Swedish GEMG Study.

OBJECTIVES: To describe Myasthenia Gravis-Activities of Daily Living (MG-ADL) in relation to clinical characteristics in a large Swedish nationwide cohort. METHODS: In a cross-sectional prevalent cohort study, the Genes and Environment in Myasthenia Gravis study (GEMG), performed November 2018 - August 2019, Myasthenia gravis (MG) patients were invited to submit an extensive 106-item life environment questionnaire, including the MG-ADL score. Patients were classified into early onset MG (EOMG, <50 years), late onset MG (LOMG, ≥50 years) or thymoma-associated MG (TAMG). Comparisons of disease-specific characteristics were made between subgroups, sex and different MG-ADL scores. RESULTS: A total of 1077 patients were included, yielding a 74% response rate: 505 (47%) were classified as EOMG, 520 (48%) LOMG and 45 (4%) TAMG. Mean age at inclusion was 64.3 years (SD 15.7) and mean disease duration was 14.6 years (SD 14.0). Complete MG-ADL scores (n=1035) ranged from 0-18p, where 26% reported a score of 0p. Higher MG-ADL scores were associated with female sex, obesity and diagnostic delay (OR=1.62, 1.72 and 1.69, P adj=0.017, 0.013 and 0.008) and inversely correlated with high educational attainment (OR=0.59, P adj=0.02), but not with age at inclusion, disease subtype nor disease duration. Almost half the population (47%) reported MG-ADL ≥3p, corresponding to an unsatisfactory symptom state. CONCLUSIONS: In this nationwide study, comprising more than 40% of the prevalent MG population in Sweden, we observe that almost half of patients report current disease symptoms associated to an unsatisfactory symptom state, indicating the need for improved treatment options.

Biologically Active Compounds in Stizolophus balsamita Inflorescences: Isolation, Phytochemical Characterization and Effects on the Skin Biophysical Parameters.

Three germacranolides, as well as five flavonoids, natural steroid and simple phenolic compounds, were isolated from the inflorescence of Stizolophus balsamita growing in Iran. The paper presents active compounds found for the first time in the inflorescence of this species. The flavonoids, simple phenolic compounds and natural steroids have been isolated for the first time in the genus Stizolophus. The MTT assay was employed to study in vitro cytotoxic effects of the taxifolin against human fibroblasts. We also evaluate the possible biological properties/cosmetic effects of Stizolophus balsamita extract and taxifolin on the human skin. Sixty healthy Caucasian adult females with no dermatological diseases were investigated. We evaluate the effects of S. balsamita extract and taxifolin on skin hydration and transepidermal water loss (TEWL). It was revealed that S. balsamita extract might decrease TEWL level and fixed the barrier function of the epidermis. The presence of bioactive phytochemical constituents in S. balsamita inflorescences makes them a valuable and safe source for creating new cosmetics and medicines.

Risk Factors of Fatigue in Idiopathic Parkinson's Disease in a Polish Population.

Introduction. Fatigue syndrome is one of the nonmotor symptoms in Parkinson's disease (PD). The aim of the study was assessment of prevalence of fatigue syndrome in PD and answering the question what are the independent risk factors connected with intensity of fatigue in PD. Methods. 114 patients with idiopathic PD (mean age 62.2 + 10.8 years) were enrolled. The fatigue was assessed according to the Fatigue Severity Scale (FSS). We analyzed associations between fatigue and sex, age, education, duration and severity of the disease, everyday activity, intensity of the main symptoms, treatment, presence of dyskinesias and fluctuations, depression and excessive sleep during the day, and presence of pain and nycturia. Results. The fatigue syndrome was detected in 57.9% of patients. The score in the FSS was 1 to 7 points, 4.3 average. Greater fatigue intensity correlated with higher total daily levodopa equivalent dose. Patients with moderate depression had significantly greater fatigue. Conclusions. Fatigue syndrome affects 57.9% of patients with PD. Use of higher LED and presence of moderate depression are independent risk factors of greater intensity of fatigue.

[Protection against tobacco smoke--compounds and substances of natural orgin]. / Ochrona przed dymem tytoniowym--zwiazki i substancje pochodzenia naturalnego.

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.

[Tobacco--a source of biofuels]. / Tyton zródlem biopaliw.

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.

[Autonomic disorders in Parkinson's disease]. / Zaburzenia autonomiczne w chorobie Parkinsona.

INTRODUCTION: Autonomic disorders (AD) are one of non-motor symptoms in Parkinson's disease (PD). The aim of this study was to assess the frequency of AD in PD patients and their relationship with age, gender, disease duration, treatment duration, L-dopa dose, and disease severity. MATERIAL AND METHODS: 101 patients with PD were recruited. Anamnesis was recorded with a focus on orthostatic hypotension, sialorrhea, dysphagia, nausea/vomiting, loss of appetite, weight loss, constipation, nycturia, urgency, pollakiuria, difficulties in starting miction, urinary incontinence, erectile dysfunction, sweating, heat/cold intolerance, and seborrhea. Mann-Whitney test and Pearson's chi2 test were used for statistical analysis. RESULTS: The study group comprised 53 men and 48 women, aged 42-84 years, mean age 67.69 years. The disease duration was 1-25 years, mean 6.5 years, L-dopa treatment duration was 1-20 years, mean 6.04 years, L-dopa dose was 300-2000 mg/24 h, mean 636 mg/24 h, UPDRS score was 8-103 pts, mean 36.38 pts. Orthostatic hypotension was found in approximately 16% of the patients. As regards gastrointestinal disorders, more than 40% of patients suffered from constipation, more than 32% from dysphagia, almost 28% from sialorrhea, approximately 11% from loss of appetite, 6.93% from weight loss, 2% from nausea/vomiting. The distribution of urogenital disorders was as follows: almost 85% of patients had erectile dysfunction, almost 57% nycturia, 24.78% urinary incontinence, approximately 22% pollakiuria, and almost 12% urgency and difficulties with erection. One-third of PD patients suffered from seborrhea, 16.83% from sweating, and almost 9% from heat/cold intolerance. CONCLUSIONS: The most frequent AD in PD were: erectile dysfunction, nycturia, constipation, dysphagia, and seborrhea. Age correlated with orthostatic hypotension, constipation, and urinary incontinence, and with erectile dysfunction in men. Women were predisposed to weight loss and sweating. Men were predisposed to difficulties in starting miction. Disease duration correlated with constipation, nycturia, and urinary incontinence; L-dopa treatment duration correlated with nausea/vomiting and constipation; L-dopa dose correlated with nausea/vomiting, constipation, urgency, and heat/cold intolerance; disease stage correlated with sialorrhea, constipation, sweating, and heat/cold intolerance.

[Tobacco cell cultures and their significance]. / Kultury komórkowe tytoniu i ich znaczenie.

Tobacco (Nicotiana tabacum L.) and related species had played a pioneer role in the development of plant biotechnology in particular in elaboration of in vitro cultures. There are several types of such cultures, of which the cell suspension cultures in liquid media are considered to be the most important for research approaches and industrial applications. Among the tobacco cell cultures the BY-2 cell system is outstanding and comparable to the HeLa human cell line. It is a unique model cell line for the study of dynamic features of plant cells. Like in the case of Arabidopsis thaliana cells, the BY-2 cell line represents a basic system for investigation of the programmed cell death in plants under influence of various factors. It is also a tool to elucide the biosynthesis of isoprenoids. Among plant cell cultures those of tobacco represent the system which is most frequently utilized for the production of recombinant proteins in which the expression of many therapeutical proteins has been achieved, such as human serum albumin, full and fragment monoclonal antibodies, interleukins, antigens and others.

[In vitro cultures of tobacco and their impact on development of plant biotechnology]. / Kultury in vitro tytoniu i ich znaczenie w rozwoju biotechnologii roslin.

Tobacco (Nicotiana tabacum) and related species had played a pioneer role in the development of plant biotechnology. Already in thirties of the XX century, aseptic in vitro cultures were established as one of the first plant tissue in vitro cultures. In the next decade the tobacco cultures served for investigations of organogenesis--formation of advetitiuos shoots and roots--the process important for micropropagation. In 60-ties of the previous century, the totipotency of plant cells, i.e. ability for regeneration of the full plant from single cell, has been unequivocally proved on the basis of experiments with tobacco cell cultures. Moreover, thanks to the tobacco in vitro cultures, the medium for performance of plant in vitro cultures was elaborated in 1962, which is the most frequently used medium till now. Tobacco was the first plant regenerated from a protoplast, as well as the first somatic hybrid and chimera in vitro were derived from tobacco cultures (the 70-ies). Tobacco was also one of the first transgenic plants and for its formation in vitro culture has been necessary to carry out transformation of tissue and regeneration of shoots and roots. Simultaneously preformed cell suspension culture of tobacco exhibited, on one hand, exceptional utility for studies of plant cell biology, and on the other hand, appeared applicable for performing controlled biosynthesis of useful metabolites and biotransformation (bioconversion), that is directed chemical modification of foreign compounds introduced to such culture. Tobacco has played the role of the model plant in investigations of biology and biotechnological applications of plants.

Assessment of rest tremor in Parkinson's disease.

BACKGROUND AND PURPOSE: Rest tremor is the most frequent sign of Parkinson's disease (PD) after bradykinesia, occurring with various severity in about 75% of patients. An objective assessment of rest tremor is difficult. The aim of the study was to analyze rest tremor in PD with the three-dimensional gauging system CMS 10; more specifically, the impact of levodopa treatment on rest tremor, the influence of clinical factors, and the correlation between rest tremor and clinical scales were assessed. MATERIAL AND METHODS: Ninety-five patients with PD (mean age 67.6 years) and 30 healthy people in a control group (mean age 59.3 years) were examined. Clinical scales (UPDRS, Hoehn and Yahr, Schwab and England, as well as Webster scale) were used to assess severity of PD. The assessment of rest tremor was performed within the more and less affected upper limb with the three-dimensional gauging system CMS 10 (Zebris GmbH) before and 1-2 hours after levodopa intake. Frequency (Hz), amplitude (deg), velocity (deg/ms) and acceleration (deg/s2) of the tremor were evaluated. Results were compared with averaged results for left and right upper limb in the control group. RESULTS: The method used in this study objectively showed asymmetry in rest tremor. After levodopa intake, all evaluated parameters of rest tremor were decreased (mainly the amplitude and frequency, and to a lesser degree, velocity and acceleration). The motor part of UPDRS showed the best correlation with rest tremor. CONCLUSIONS: The three-dimensional measuring system CMS 10 is useful in the objective assessment of rest tremor in PD. Rest tremor in PD is under the influence of PD form, the intake of levodopa dose, the amount of levodopa, gender and level of education.

Neurological syndromes in systemic lupus erythematosus and their association with antiphospholipid syndrome.

BACKGROUND AND PURPOSE: In patients with systemic lupus erythematosus (SLE), severe neurological syndromes indicating central, peripheral or autonomic nervous system involvement are frequently seen. Antiphospholipid syndrome (APS) increases the risk of nervous system involvement. The aim of the study was to assess neurological manifestations occurring in patients with SLE and to evaluate their association with APS. MATERIAL AND METHODS: One hundred thirty-seven patients (123 women and 14 men) with SLE were studied. Fifty out of 137 patients (43 women and 7 men) were diagnosed with APS. All patients underwent full neurological examination and diagnostics. Neurological syndromes were classified according to the standardized American College of Rheumatology (ACR) nomenclature. RESULTS: Neurological syndromes were found in 89 cases out of 137 admitted SLE patients (64.96%). Headache was present in 52 patients (37.96%), polyneuropathy in 17 (12.41%), cerebrovascular disease in 13 (9.49%), seizures in 10 (7.3%), cranial neuropathy in 5 (3.65%), demyelinating syndrome in 5 (3.65%), mononeuropathy in 3 (2.19%), movement disorder in 3 (2.19%), and aseptic meningitis in 2 patients (1.46%). Our study indicated that 78% of patients with APS showed nervous system involvement as compared with 57.47% of SLE patients without APS (p=0.01). CONCLUSIONS: Neurological syndromes occurring most frequently in SLE patients include headache, polyneuropathy and cerebrovascular diseases. Antiphospholipid syndrome increases the risk of nervous system involvement. We found that APS was strongly associated with neurological manifestations and in particular with cerebrovascular diseases and seizures.

Phenylethanoid glucosides from in vitro propagated plants and callus cultures of Plantago lanceolata.

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.

[Clinical correlation of vascular parkinsonism]. / Korelacje kliniczne parkinsonizmu naczyniowego.

Vascular parkinsonism has not been well defined and the clinical correlation of vascular parkinsonism is still not clear. The aim of the study was to estimate prevalence of occurrence of vascular parkinsonism, analysis of risk factors leading to its development and to identify clinical features that suggest a vascular origin. 214 patients with Parkinson's disease were examined. Their ages ranged from 37 to 88 years (median 66.4 years). Evidence of vascular parkinsonism was assessed using a vascular rating scale previously described by Winikates and Jankovic. Statistical analysis was performed with Mann-Whitney U test, chi 2 Pearson test, chi 2 Yates test, Spearman rank correlation and Student's t test. Out of 214 patients 8 were proved to have developed Parkinson's disease due to vascular disease, what gave 3.74%. Out of risk factors for stroke 5 patients had hypertension, 3 had diabetes mellitus, 2 suffered from heart disease, 2 had infarctus myocardii, 1 had hyperlipidemia, 1 had atrial fibrillation. Additionally, those patients had neuroimaging (CT or MRI) evidence of vascular disease in one or more vascular territories. Patients with vascular parkinsonism were older, had shorter duration of disease, were more likely to present rigidity rather than tremor. Dementia and incontinence were more common in vascular group than in Parkinson's disease group. Patients with vascular parkinsonism were also significantly more likely to have corticospinal findings. Proving that Parkinson's disease had vascular etiology is extremely difficult. The test results are inconclusive.

Naphthalene glucoside and other phenolics from the shoot and callus cultures of Drosophyllum lusitanicum.

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.