Papers by Wojciech Szypuła
Acta Biologica Cracoviensia. Series Botanica. Supplement, 2009
12th National Conference'In vitro Cultures, Poznań 2009' 66 POSTERS ACTA BIOLO... more 12th National Conference'In vitro Cultures, Poznań 2009' 66 POSTERS ACTA BIOLOGICA CRACOVIENSIA Series Botanica Gametophytes as well as sporophytes of H. selago are col-onized by endophytic fungi (Higgins et al., 2007; Winther and Friedman 2008). The class Dothideomycetes is ...
Reference series in phytochemistry, 2023
Extracts of Rhodiola rosea obtained from intact plant roots harvested from the Medicinal Plants G... more Extracts of Rhodiola rosea obtained from intact plant roots harvested from the Medicinal Plants Garden of the Institute of Medicinal Plants in Poznan, from plants growing in the wild (Babia G6ra in the Carpathian Mountains) and from callus cultivated in vitro reduced the growth of the three Staphylococcus aureus strains: ATCC 6538 P, ATCC 25923 and NCTC 4163. S. aureus NCTC 4163 was the most sensitive to their action. The extracts of intact plant roots produced a higher growth inhibition zone than callus extracts.
Biotechnologia. Journal of Biotechnology, Computational Biology and Bionanotechnology, 2015
Tissue culture was developed at the beginning of the twentieth century (1907) as a method for stu... more Tissue culture was developed at the beginning of the twentieth century (1907) as a method for studying the behaviour of human and animal cells free of systemic variations that might arise in vivo both during normal homeostasis and under the stress. The technique was established first with disaggregated fragments of tissue, and growth was restricted to the radial migration of cells from the tissue fragment, with occasional mitoses in the outgrowth. As culture of cells from and within such primary explants of tissue dominated the field for more than 50 years, it is not surprising that the name "tissue culture" has remained in use as a generic term despite the fact that most of the expansion in the field in the second half of the twentieth century was made possible by the use of dispersed cell cultures. Disaggregation of explanted cells and subsequent plating out of the dispersed cells was first demonstrated by Rous (Rous and Jones, 1916). In 1950s trypsin became more generally used for subculture, following procedures described by Dulbecco to obtain passaged monolayer cultures for viral plaque assays (Dulbecco, 1952), and the generation of a single cell suspension by trypsinization. Tissue culture enter its second century since its inception. For the first time it is possible for genetic manipulation of commonly and easily cultured cells, such as skin fibroblasts, to allow their conversion into pluripotent stem cells (iPS), capable of differentiating into a range of different cell types. The field opens up to a whole new scenario: instead of the need for complex selective culture technique, the culture procedures may be used to initiate biochemical regulation and may be used to convert phenotype. Tissue culturing nowadays is essential and irreplaceable in modern biotechnology as a method supplementing and enriching our knowledge. Without specialized laboratories for tissue culture with equipment for they analysis and chance of cytogenetic and molecular analysis it will not be possible to by us transgenic rabbits producing in mammary gland human growth hormone and five lines of transgenic pigs for xenotransplantation purposes. In our laboratories we use cell culture of human and animal cells for biotechnology needs or as models. In animal transgenesis we usually perform several steps using molecular and cytogenetic methods at cell culture levels. Important issues in producing transgenic animals cover characteristics of donor of gene, transfection and cell transformation, detection of transgene, mapping of transgene, passing of transgene into next generation, homozygote selection and analysis of transgene activity. From cytogenetic methods the most important are banding chromosomes methods for karyotyping each transgenic individual and mapping the transgene on metaphase chromosomes using fluorescent in situ hybridization (FISH). It is possible to study structure of the transgene on the chromatin level using FISH on chromosomal loop domain or using 3D FISH technique. This methods is important to distinguish homozygous offspring from heterozygous and confirm the stability of transgenesis. The last step include analysis of expression of transgene using flow cytometry methods.
Polish Journal of Ecology, 2010
The aim of the study was to assess the influence of environmental conditions on the composition o... more The aim of the study was to assess the influence of environmental conditions on the composition of endophytic assemblages associated with fir clubmoss, Huperzia selago (L.) Bernh. ex Schrank et Mart. This clubmoss is a cosmopolitan species widely spread in Eurasia and North America, and its gametophytes and sporophytes are colonised by endophytic fungi. Plants for this study were collected in October 2004 and between April and October 2007 from seven European sites, differing in altitude, average annual precipitation, temperature and plant community. Endophytes were isolated on PDA medium (4% potato-dextrose agar) and identified through morphological observations. Different ecological indices were calculated for each site like colonization factor, Fisher’s alpha, Shannon’s diversity, Morista-Horn and Jaccard’s indices. Our results prove that altitude positively correlates with species diversity (Fisher’s alpha) and species richness (number of species). Moreover, the similarity asses...
ABSTRACT Effects of hydrodynamic turbulent stress on disintegration of plant cell aggregates Rhod... more ABSTRACT Effects of hydrodynamic turbulent stress on disintegration of plant cell aggregates Rhodiola rosea L.(Crassulaceae) were investigated. Investigations were carried out in the stirred tank mixer equipped with the Rushton type turbine mixer and four baffles under conditions when breakage of cell aggregates dominates the process. Breakage process was interpreted with the multifractal model of turbulence.
Molecules
This is the first report of an efficient and effective procedure to optimize the biosynthesis of ... more This is the first report of an efficient and effective procedure to optimize the biosynthesis of huperzine A (HupA) and huperzine B (HupB) in vitro from Huperzia selago gametophytes. Axenic tissue cultures were established using spores collected from the sporophytes growing in the wild. The prothalia were obtained after 7–18 months. Approximately 90 up to 100% of the gametophytes were viable and grew rapidly after each transfer on to a fresh medium every 3 months. The best biomass growth index for prothallus calculated on a fresh (FW) and dry weight (DW) basis, at 24 weeks of culture, was 2500% (FW) and 2200% (DW), respectively. The huperzine A content in the gametophytes was very high and ranged from 0.74 mg/g to 4.73 mg/g DW. The highest yield HupA biosynthesis at >4 mg/g DW was observed on W/S medium without growth regulators at 8 to 24 weeks of culture. The highest HupB content ranged from 0.10 mg/g to 0.52 mg/g DW and was obtained on the same medium. The results demonstrate ...
Folia Neuropathologica, 2016
Imbalance between production and scavenging of free radicals and other reactive oxygen species (R... more Imbalance between production and scavenging of free radicals and other reactive oxygen species (ROS) is a component of many diseases, but it is especially important in aging-related diseases of the central nervous system. Oxidative stress-induced neuronal dysfunction plays an important role in the pathomechanism of neurodegenerative disorders, including Alzheimer's and Parkinson's disease. Experimental data showed that free radical scavengers may protect the brain against oxidative modifications. The need for efficient and safe antioxidants with therapeutic potential stimulated the rise of interest in the medicinal plant products, which are a rich source of phytochemicals possessing biological activity. In our studies we focused on alkaloid fractions (AFs) isolated from club moss, Huperzia selago and Diphasiastrum complanatum, due to their beneficial activity and exclusive chemical structure. Our previous study demonstrated that selected alkaloids from Huperzia selago effectively protect macromolecules from oxidative damage. Therefore, in the present study we investigated the effects and mechanisms of action of AFs isolated from Huperzia selago and Diphasiastrum complanatum against sodium nitroprusside (SNP)-induced oxidative injury in PC12 cells. The results demonstrated that the selected AFs via reduction of nitric oxide (NO) liberation protected cells against oxidative stress, DNA and mitochondrial damage, as well as apoptosis caused by SNP. Selected AF notably decreased SNP-evoked mitochondrial polymerase γ (Polg) up-regulation. Furthermore, AF which contains Lycopodine, Serratidine, Lycoposerramine-G and (probably) Cermizine B completely inhibited the SNP-induced expression of interferon-γ (Ifng) and cyclooxygenase 2 (Ptgs2) as well as significantly down-regulated the expression of 12/15-lipoxygenase (Alox12) and tended to decrease the mRNA level of interleukin-6 gene (Il6). In conclusion, these results suggest that the AFs from Huperzia selago effectively protect PC12 cells against SNP-induced oxidative damage by adjusting the level of reactive nitrogen species, suppression of apoptosis and down-regulation of proinflammatory genes. The compounds present in these AFs could be potential candidates to develop successful drugs preventing oxidative damage and apoptosis in age-related neurodegenerative disorders.
Mycotaxon Ithaca Ny, 2011
Paraconiothyrium is a recently established genus within the order Pleosporales. Species from this... more Paraconiothyrium is a recently established genus within the order Pleosporales. Species from this genus are commonly associated with plants but can also be found in soil samples and be parasitic on fungi. Several isolates of a Paraconiothyrium sp. were obtained from Huperzia selago in Poland. Strains were characterized based on morphological characteristics and molecular data (SSU rDNA, ITS1, 5.8S rDNA, ITS2). Based on its unique morphology and DNA phylogeny, isolates were described as a new species: Paraconiothyrium babiogorense, which represents the first report of a Paraconiothyrium species from fir club moss. Comparison of characters with other Paraconiothyrium species is provided as well as a signature sequence for the new species.
Polish Journal of Ecology, 2010
The aim of the study was to assess the influence of environmental conditions on the composition o... more The aim of the study was to assess the influence of environmental conditions on the composition of endophytic assemblages associated with fir clubmoss, Huperzia selago (l.) Bernh. ex. Schrank et Mart. This clubmoss is a cosmopolitan species widely spread in Eurasia and north America, and its gametophytes and sporophytes are colonised by endophytic fungi. Plants for this study were collected between April and october 2007 from seven European sites, differing in altitude, average annual precipitation, temperature and plant community. Endophytes were isolated on PDA medium (4% potato-dextrose agar) and identified through morphological observations. Different ecological indices were calculated for each site like colonization factor, fisher's alpha, Shannon's diversity, Morista-horn and Jaccard's indices. our results prove that altitude positively correlates with species diversity (fisher's alpha) and species richness (number of species). Moreover, the similarity assessed from Jaccard's (0.66 for larici-piceetum sites) and Morista-horn indices (0.77 for Juncetea trifidi sites) suggest the influence of plant community on fungal endophyte composition.
Inzynieria Chemiczna i Procesowa
Effects of hydrodynamic turbulent stress on disintegration of plant cell aggregates Rhodiola rose... more Effects of hydrodynamic turbulent stress on disintegration of plant cell aggregates Rhodiola rosea L.(Crassulaceae) were investigated. Investigations were carried out in the stirred tank mixer equipped with the Rushton type turbine mixer and four baffles under conditions when breakage of cell aggregates dominates the process. Breakage process was interpreted with the multifractal model of turbulence.
Inzynieria Chemiczna i Procesowa
Interest in the taxonomy of clubmosses has substantially increased in recent years and together w... more Interest in the taxonomy of clubmosses has substantially increased in recent years and together with these studies, efforts towards a correct classification of this ancient group of plants have also progressed. Some taxonomic changes within Lycopodiales since the first edition of Species Plantarum (Linnaei 1753) to the contemporary papers based on molecular phylogeny are summarized. Selected problems from the taxonomy and nomenclature of some groups of the order Lycopodiales are discussed in this paper.
Botanical Guidebooks (continuation of Polish Botanical Studies, Guidebook Series) is an irregular... more Botanical Guidebooks (continuation of Polish Botanical Studies, Guidebook Series) is an irregularly issued series publishing guides to Polish botanical institutions, collections and collectors, botanical literaiure, and directories as well as botanical field-guides and basic information regarding various branches of botany.
W Karpatach polskich i w Polsce gatunek narażony na wyginięcie (VU)-Cz. K. Uwagi taksonomiczne Ca... more W Karpatach polskich i w Polsce gatunek narażony na wyginięcie (VU)-Cz. K. Uwagi taksonomiczne Campanula serrata (Kit.) Hendrych jest zróżnicowana na dwa podgatunki: typowy subsp. serrata i subsp. recta (Dulac) Podlech. W Polsce rośnie wyłącznie podgatunek typowy. Morfologia i biologia Hemikryptofi t, bylina do 80 cm wysokości, z krótkim kłączem i korzeniem burakowato zgrubiałym. Tworzy płonne różyczki liściowe oraz pojedyncze lub wyrastające po kilka pędy kwiatowe. Korona ciemnoniebieskofi oletowa, dzwonkowata, długości 1,5-2 cm, rozcięta na 5 łatek. Owocem jest torebka odwrotnie jajowata, otwierająca się 3 dziurkami blisko nasady. Liczba chromosomów 2n = 34 (Kovanda 1970)-dane spoza Polski. Kwitnie od lipca do września. Jest rośliną owadopylną. Nektar gromadzi się u nasady szyjki słupka. Nasiona z mięsistym bielmem, liczne, drobne, bez przystosowań do rozsiewania. Zasięg ogólny Endemit ogólnokarpacki (Pawłowski 1970b; Piękoś-Mirkowa i in. 1996b). Rośnie w Karpatach słowackich, polskich, ukraińskich i rumuńskich. Rozmieszczenie w Polsce Występuje w kilku pasmach karpackich (Beskid Żywiecki, Tatry, Gorce, Bieszczady) (Piękoś-Mirkowa 2004). Rozmieszczenie w Karpatach polskich Dzwonek piłkowany jest gatunkiem wysokogórskim (subalpejskim). W Tatrach rośnie na kilkunastu stanowiskach rozmieszczonych od 920 m (Siwa Polana) do 1950 m n.p.m. (Jarząbczy Wierch-Kamienista) (
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Papers by Wojciech Szypuła