Papers by Marta Palusinska-szysz
Frontiers in microbiology, Jan 31, 2024
Archivum Immunologiae Et Therapiae Experimentalis, Jul 4, 2009
The Legionellae are Gram-negative bacteria able to survive and replicate in a wide range of proto... more The Legionellae are Gram-negative bacteria able to survive and replicate in a wide range of protozoan hosts in natural environments, but they also occur in man-made aquatic systems, which are the major source of infection. After transmission to humans via aerosols, Legionella spp. can cause pneumonia (Legionnaires' disease) or influenza-like respiratory infections (Pontiac fever). In children, Legionnaires' disease is uncommon and is mainly diagnosed in children with immunosuppression. The clinical picture of Legionella pneumonia does not allow differentiation from pneumonia caused by others pathogens. The key to diagnosis is performing appropriate microbiological testing. The clinical presentation and the natural course of Legionnaires' disease in children are not clear due to an insufficient number of samples, but morbidity and mortality caused by this infection are extremely high. The mortality rate for legionellosis depends on the promptness of an appropriate antibiotic therapy. Fluoroquinolones are the most efficacious drugs against Legionella. A combination of these drugs with macrolides seems to be promising in the treatment of immunosuppressed patients and individuals with severe legionellosis. Although all Legionella species are considered potentially pathogenic for humans, Legionella pneumophila is the etiological agent responsible for most reported cases of community-acquired and nosocomial legionellosis.
Postȩpy higieny i medycyny doświadczalnej, 2006
Legionellae are Gram-negative obligate intracellular parasites of unicellular animal organisms, s... more Legionellae are Gram-negative obligate intracellular parasites of unicellular animal organisms, some of which are sometimes able to cause an acute and sever pneumonia in humans. Legionellae, are capable of surviving for long periods in water when between hosts. Their adaptation to these quite diverse environments seems to be accomplished by morphological and biochemical pathway changes. It has been well established that humans coexisted with these bacteria for a long period of time, and it is only recently that our industrial technology provided these organisms with the means of causing infection. This review describes the invasion process and the life cycle of Legionellae in both protozoan and mammalian cells. Our understanding of the infection cycle of Legionellae is primarily based on observations by transmission and scanning EM and by contrast phase and fluorescence microscopy. The identification of virulence determinants by molecular techniques is also discussed. The bacteria enter the host cell mainly by coiling phagocytosis and reside within unique phagosomes which, during the first hours of infection, are isolated from the endosomal pathway. Within the protected vacuole the mature infectious forms of Legionellae convert to replicative forms that no longer express virulence traits. Evidence is provided on how the biogenesis of the replication niche is determined. The virulence factors that arrest phagosome maturation during intracellular replication are also described. The status of our current knowledge on the means by which Legionellae successfully infect the host cells to cause disease is discussed.
Naturalnym środowiskiem występowania bakterii z rodziny Legionellaceae jest woda o różnym zakresi... more Naturalnym środowiskiem występowania bakterii z rodziny Legionellaceae jest woda o różnym zakresie temperatury, pH, zawartości tlenu i substancji odżywczych. Zdolność do przetrwania w zmieniających się fi zyko-chemicznych warunkach w wodzie umożliwia im związek z innymi mikroorganizmami, z którymi tworzą biofi lm. Wewnątrzkomórkowy tryb życia w określonych gatunkach pierwotniaków zapewnia Legionellaceae nie tylko stały dopływ substancji odżywczych, umożliwiający im intensywne podziały, ale również ochronę przed działaniem bakteriobójczych środków dezynfekcyjnych. Bakterie po przedostaniu się do sztucznych systemów dystrybucji wody stają się źródłem niebezpiecznego dla zdrowia człowieka wodno-powietrznego aerozolu. Legionellae odpowiedzialne są zarówno za grypopodobne infekcje (gorączka Pontiac), ciężką postać zapalenia płuc, jak i pozapłucne umiejscowienie choroby, szczególnie u osób z supresją immunologiczną. Pacjenci z chorobą legionistów wymagają szybkiej interwencji medycznej. Lekiem z wyboru okazała się erytromycyna. Obecnie w leczeniu legionelozy stosuje się nowsze makrolidy oraz fl uorochinolony. Spośród 52 opisanych gatunków zaliczonych do rodziny Legionellaceae, Legionella pneumophila w tym serotyp 1 jest najczęstszym czynnikiem wywołującym legionelozowe zapalenie płuc. Bakteria ta odpowiada za 80% przypadków legionelozy nabytej zarówno w środowisku zamieszkania jak i zakażeń szpitalnych.
Journal of Applied Genetics, Sep 30, 2015
Legionella bacteria are organisms of public health interest due to their ability to cause pneumon... more Legionella bacteria are organisms of public health interest due to their ability to cause pneumonia (Legionnaires' disease) in susceptible humans and their ubiquitous presence in water supply systems. Rapid diagnosis of Legionnaires' disease allows the use of therapy specific for the disease. L. pneumophila serogroup 1 is the most common cause of infection acquired in community and hospital environments. The non-L. pneumophila infections are likely under-detected because of a lack of effective diagnosis. In this work, simplex and duplex PCR assays with the use of new molecular markers pcs and pmtA involved in phosphatidylcholine synthesis were specified for rapid and cost-efficient identification and distinguishing Legionella species. The sets of primers developed were found to be sensitive and specific for reliable detection of Legionella belonging to the eight most clinically relevant species. Among these, four primer sets I, II, VI, and VII used for duplex-PCRs proved to have the highest identification power and reliability in the detection of the bacteria. Application of this PCR-based method should improve detection of Legionella spp. in both clinical and environmental settings and facilitate molecular typing of these organisms.
Biochimica Et Biophysica Acta - Biomembranes, Oct 1, 2014
The greater wax moth Galleria mellonella has been exploited worldwide as an alternative model hos... more The greater wax moth Galleria mellonella has been exploited worldwide as an alternative model host for studying pathogenicity and virulence factors of different pathogens, including Legionella pneumophila, a causative agent of a severe form of pneumonia called Legionnaires' disease. An important role in the insect immune response against invading pathogens is played by apolipophorin III (apoLp-III), a lipid-and pathogen associated molecular pattern-binding protein able to inhibit growth of some Gram-negative bacteria, including Legionella dumoffii. In the present study, anti-L. pneumophila activity of G. mellonella apoLp-III and the effects of the interaction of this protein with L. pneumophila cells are demonstrated. Alterations in the bacteria cell surface occurring upon apoLp-III treatment, revealed by Fourier transform infrared (FTIR) spectroscopy and atomic force microscopy, are also documented. ApoLp-III interactions with purified L. pneumophila LPS, an essential virulence factor of the bacteria, were analysed using electrophoresis and immunoblotting with anti-apoLp-III antibodies. Moreover, FTIR spectroscopy was used to gain detailed information on the type of conformational changes in L. pneumophila LPS and G. mellonella apoLp-III induced by their mutual interactions. The results indicate that apoLp-III binding to components of bacterial cell envelope, including LPS, may be responsible for anti-L. pneumophila activity of G. mellonella apoLp-III.
Microbiological Research, Feb 1, 2011
The phospholipid class and fatty acid composition of Legionella bozemanae were determined using t... more The phospholipid class and fatty acid composition of Legionella bozemanae were determined using thin-layer chromatography, gas-liquid chromatography, and matrix-assisted laser desorption ionization-time of flight mass spectrometry. Phosphatidylcholine, phosphatidylethanolamine, and diphosphatidylglycerol were the predominant phospholipids, while phosphatidyl-N-monomethylethanolamine, phosphatidylglycerol, and phosphatidyl-N,N-dimethylethanolamine were present at low concentrations. With the use of the LC/MS technique, PC16:0/15:0, PC17:/15:0, and PE16:1/15:0 were shown to be the dominant phospholipid constituents, which may be taxonomically significant. Two independent phosphatidylcholine synthesis pathways (the three-step methylation and the one-step CDP-choline pathway) were present and functional in L. bozemanae. In the genome of L. bozemanae, genes encoding two potential phosphatidylcholine forming enzymes, phospholipid N-methyl transferase (PmtA) and phosphatidylcholine synthase (Pcs), homologous to L. longbeachae, L. drancourtii, and L. pneumophila pmtA and pcs genes were identified. Genes pmtA and pcs from L. bozemanae were sequenced and analyzed on nucleotide and amino acid levels. Bacteria grown on an artificial medium with labelled choline synthesized phosphatidylcholine predominantly via the phosphatidylcholine synthase pathway, which indicates that L. bozemanae phosphatidylcholine, similarly as in other bacteria associated with eukaryotes, is an important determinant of host-microbe interactions.
Recent Patents on Anti-infective Drug Discovery, Jun 1, 2009
Acta Biochimica Polonica, Mar 20, 2014
Among Legionella species, which are recognized to be pathogenic for humans, L. gormanii is the se... more Among Legionella species, which are recognized to be pathogenic for humans, L. gormanii is the second prevalent causative agent of community-acquired pneumonia after L. pneumophila. Anti-L. gormanii activity of Galleria mellonella hemolymph extract and apolipophorin III (apoLp-III) was examined. The extract and apoLp-III at the concentration 0.025 mg/ml caused 75% and 10% decrease of the bacteria survival rate, respectively. The apoLp-III-induced changes of the bacteria cell surface were analyzed for the first time by atomic force microscopy. Our studies demonstrated the powerful anti-Legionella effects of the insect defence polypeptides, which could be exploited in drugs design against these pathogens.
Molecules, Jan 6, 2022
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Systematic and Applied Microbiology, 2001
Legionella lytica comb. nov. an intracellular bacterial pathogen of small free-living amoebae was... more Legionella lytica comb. nov. an intracellular bacterial pathogen of small free-living amoebae was subjected to cellular fatty acid (FA) analysis employing base and acid catalyzed cleavage, gas-liquid chromatography and mass spectrometry. Both unbranched and branched (iso and anteiso) FA of chains ranging from 14 to 30 carbon atoms occurred. The presence of two long-chain FA: 27-oxo-octacosanoic acid and heptacosane-1,27-dioic acid, characteristic for legionellae, was found. Nine amide-linked 3-hydroxy-FA were revealed. The main 3-hydroxy-fatty acids comprise: 3-OH-14:0, 3-OH-16:0, 3-OH-18:0, 3-OH-i18:0, 3-OH-15:OH, 3-OH-i16:0 amd 3-OH-i17:0. The profile of hydroxy FAs permits allocation of L. lytica to group 3 of legionellae which comprise blue-white fluorescent species.
International Journal of Molecular Sciences, May 9, 2014
Phosphatidycholine (PC) is the major membrane-forming phospholipid in eukaryotes but it has been ... more Phosphatidycholine (PC) is the major membrane-forming phospholipid in eukaryotes but it has been found in only a limited number of prokaryotes. Bacteria synthesize PC via the phospholipid N-methylation pathway (Pmt) or via the phosphatidylcholine synthase pathway (Pcs) or both. Here, we demonstrated that Legionella dumoffii has the ability to utilize exogenous choline for phosphatidylcholine (PC) synthesis when bacteria grow in the presence of choline. The Pcs seems to be a primary pathway for synthesis of this phospholipid in L. dumoffii. Structurally different PC species were distributed in the outer and inner membranes. As shown by the LC/ESI-MS analyses,
Frontiers in Microbiology, Dec 17, 2019
The Legionella pneumophila TF3/1 mutant of the Corby strain, which possesses a point mutation in ... more The Legionella pneumophila TF3/1 mutant of the Corby strain, which possesses a point mutation in the active site of the O-acetyltransferase, synthesized the polysaccharide chain with a reduced degree of substitution with O-acetyl groups. The mutant did not produce a high-molecular-weight lipopolysaccharide (LPS) fraction above 12 kDa. The disturbances in LPS synthesis have an effect on the composition of other macromolecules (lipids and proteins), as indicated by differences in the infrared absorption spectra between the L. pneumophila Corby strain and its TF3/1 mutant. The wild-type strain contained less N +-CH 3 and C-N groups as well as more CH 3 groups than the mutant. The fatty acid composition showed that the wild type strain synthesized more branched acyl residues (a15:0, i16:0, and a17:0), a less unsaturated acid (16:1), and a straight-chain acid (18:0) than the mutant. The mutant synthesized approximately twice more a long-chain fatty acid (20:0) than the wild type. The main differences in the phospholipids between both strains were found in the classes of phosphatidylcholines and phosphatidylglycerols (PG). Substantial differences in the cell surface topography of these bacteria and their nanomechanical properties were shown by atomic force microscopy (AFM). The wild type strain had no undulated surface and produced numerous vesicles. In the case of the mutant type, the vesicles were not numerous, but there were grooves on the cell surface. The average roughness of the cell surface of the mutant was approximately twofold higher than in the wild-type strain. In turn, the wild-type strain exhibited much better adhesive properties than the mutant. The kinetic study of the interaction between the L. pneumophila strains and Acanthamoeba castellanii monitored by Förster resonance energy transfer revealed a pronounced difference, i.e., almost instantaneous and highly efficient binding of the L. pneumophila Corby strain to the amoeba surface, followed by penetration into the
International Journal of Molecular Sciences, Feb 2, 2021
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Postȩpy higieny i medycyny doświadczalnej, 2021
Bakterie Legionella pneumophila w środowisku naturalnym pasożytują wewnątrz komórek wybranych gat... more Bakterie Legionella pneumophila w środowisku naturalnym pasożytują wewnątrz komórek wybranych gatunków pierwotniaków, a po przedostaniu się do sztucznych systemów dystrybucji wody stają się ważnym czynnikiem etiologicznym zapalenia płuc u ludzi. Główną cechą determinującą patogenność tych bakterii jest zdolność do życia i replikacji w makrofagach płucnych, czyli w komórkach wyspecjalizowanych do fagocytozy, zabijania i trawienia mikroorganizmów. Warunkiem wstępnym rozwoju infekcji jest przełamanie mechanizmów bójczych makrofagów i utworzenie wakuoli replikacyjnej LCV (Legionella containing vacuole). Biogeneza wakuoli LCV jest możliwa dzięki sprawnemu funkcjonowaniu IV systemu sekrecji Dot/Icm, który jest wielobiałkowym, złożonym kompleksem umiejscowionym w wewnętrznej i zewnętrznej membranie osłony komórkowej bakterii. System Dot/Icm liczy 27 elementów, na które składają się m.in. kompleks rdzeniowo-transmembranowy, tworzący strukturalny szkielet całego systemu oraz kompleks białek sprzęgających. Geny kodujące komponenty systemu Dot/Icm są zorganizowane na dwóch regionach chromosomu bakteryjnego. System sekrecji Dot/Icm umożliwia L. pneumophila wprowadzenie do cytozolu komórki gospodarza ponad 300 białek efektorowych, których skoordynowane działanie powoduje utrzymanie integralności błony wakuoli replikacyjnej oraz pozwala na manipulowanie różnymi procesami komórki. Ważnym elementem strategii wewnątrzkomórkowego namnażania się L. pneumophila jest modulowanie transportu pęcherzykowego, interakcja z retikulum endoplazmatycznym oraz zakłócenie biosyntezy białek, procesów autofagii i apoptozy komórki gospodarza. Poznanie złożonych mechanizmów regulacji i funkcji białek efektorowych systemu Dot/Icm ma decydujące znaczenie w zapobieganiu i leczeniu choroby legionistów.
Sustainability, Apr 14, 2023
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Membranes, Mar 20, 2023
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Metabolites, May 6, 2022
Unusual Lipid Components of Legionella gormanii Membranes.
International Journal of Molecular Sciences, Dec 12, 2012
The gram-negative bacterium Legionella dumoffii is, beside Legionella pneumophila, an etiological... more The gram-negative bacterium Legionella dumoffii is, beside Legionella pneumophila, an etiological agent of Legionnaires' disease, an atypical form of pneumonia. The aim of this study was to determine the antimicrobial activity of Galleria mellonella defense polypeptides against L. dumoffii. The extract of immune hemolymph, containing a mixture of defense peptides and proteins, exhibited a dose-dependent bactericidal effect on L. dumoffii. The bacterium appeared sensitive to a main component of the hemolymph extract, apolipophorin III, as well as to a defense peptide, Galleria defensin, used at the concentrations 0.4 mg/mL and 40 μg/mL, respectively. L. dumoffii cells cultured in the presence of choline were more susceptible to both defense factors analyzed. A transmission electron microscopy study of bacterial cells demonstrated that Galleria defensin and apolipophorin III induced irreversible cell wall damage and strong intracellular alterations, i.e., increased vacuolization, cytoplasm
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Papers by Marta Palusinska-szysz