Application of molecular techniques for the assessment of microorganism diversity on cultural heritage objects

As a result of their unpredictable ability to adapt to varying environmental conditions, microorganisms inhabit different types of biological niches on Earth. Owing to the key role of microorganisms in many biogeochemical processes, trends in modern microbiology emphasize the need to know and understand the structure and function of complex microbial communities. This is particularly important if the strategy relates to microbial communities that cause biodeterioration of materials that constitute our cultural heritage. Until recently, the detection and identification of microorganisms inhabiting objects of cultural value was based only on cultivation-dependent methods. In spite of many advantages, these methods provide limited information because they identify only viable organisms capable of growth under standard laboratory conditions. However, in order to carry out proper conservation and renovation, it is necessary to know the complete composition of microbial communities and their activity. This paper presents and characterizes modern techniques such as genetic fingerprinting and clone library construction for the assessment of microbial diversity based on molecular biology. Molecular methods represent a favourable alternative to culture dependent methods and make it possible to assess the biodiversity of microorganisms inhabiting technical materials and cultural heritage objects

Metabolomic and high-throughput sequencing analysis—modern approach for the assessment of biodeterioration of materials from historic buildings

Preservation of cultural heritage is of paramount importance worldwide. Microbial colonization of construction materials, such as wood, brick, mortar and stone in historic buildings can lead to severe deterioration. The aim of the present study was to give modern insight into the phylogenetic diversity and activated metabolic pathways of microbial communities colonized historic objects located in the former Auschwitz II-Birkenau concentration and extermination camp in Oświęcim, Poland. For this purpose we combined molecular, microscopic and chemical methods. Selected specimens were examined using Field Emission Scanning Electron Microscopy (FESEM), metabolomic analysis and high-throughput Illumina sequencing. FESEM imaging revealed the presence of complex microbial communities comprising diatoms, fungi and bacteria, mainly cyanobacteria and actinobacteria, on sample surfaces. Microbial diversity of brick specimens appeared higher than that of the wood and was dominated by algae and cyanobacteria, while wood was mainly colonized by fungi. DNA sequences documented the presence of 15 bacterial phyla representing 99 genera including Halomonas, Halorhodospira, Salinisphaera, Salinibacterium, Rubrobacter, Streptomyces, Arthrobacter and 9 fungal classes represented by 113 genera including Cladosporium, Acremonium, Alternaria, Engyodontium, Penicillium, Rhizopus and Aureobasidium. Most of the identified sequences were characteristic of organisms implicated in deterioration of wood and brick. Metabolomic data indicated the activation of numerous metabolic pathways, including those regulating the production of primary and secondary metabolites, for example, metabolites associated with the production of antibiotics, organic acids and deterioration of organic compounds. The study demonstrated that a combination of electron microscopy imaging with metabolomic and genomic techniques allows to link the phylogenetic information and metabolic profiles of microbial communities and to shed new light on biodeterioration processes

Targeting membrane-bound viral RNA synthesis reveals potent inhibition of diverse coronaviruses including the middle East respiratory syndrome virus.

Coronaviruses raise serious concerns as emerging zoonotic viruses without specific antiviral drugs available. Here we screened a collection of 16671 diverse compounds for anti-human coronavirus 229E activity and identified an inhibitor, designated K22, that specifically targets membrane-bound coronaviral RNA synthesis. K22 exerts most potent antiviral activity after virus entry during an early step of the viral life cycle. Specifically, the formation of double membrane vesicles (DMVs), a hallmark of coronavirus replication, was greatly impaired upon K22 treatment accompanied by near-complete inhibition of viral RNA synthesis. K22-resistant viruses contained substitutions in non-structural protein 6 (nsp6), a membrane-spanning integral component of the viral replication complex implicated in DMV formation, corroborating that K22 targets membrane bound viral RNA synthesis. Besides K22 resistance, the nsp6 mutants induced a reduced number of DMVs, displayed decreased specific infectivity, while RNA synthesis was not affected. Importantly, K22 inhibits a broad range of coronaviruses, including Middle East respiratory syndrome coronavirus (MERS-CoV), and efficient inhibition was achieved in primary human epithelia cultures representing the entry port of human coronavirus infection. Collectively, this study proposes an evolutionary conserved step in the life cycle of positive-stranded RNA viruses, the recruitment of cellular membranes for viral replication, as vulnerable and, most importantly, druggable target for antiviral intervention. We expect this mode of action to serve as a paradigm for the development of potent antiviral drugs to combat many animal and human virus infections

Cabaret as a community

Praca dotyczy antropologicznego wymiaru kabaretu jako wspólnoty. W rozdziale pierwszym (Fenomen kabaretu) kabaret charakteryzowany jest ze względu na cechy wyodrębniające go spośród wszystkich dziedzin sztuki, które decydują o tym, że może być nazwany sztuką śmiechu. Rozdział drugi (Kabaret w Polsce) prezentuje najważniejsze kabarety w Polsce, z uwzględnieniem ich wpływu na rozwój tej sztuki. W rozdziale trzecim (Mechanizmy kształtujące współczesny kabaret polski) poruszone zostają kwestie, które znacząco wpływają na obraz współczesnego kabaretu w Polsce oraz pojawiają się głosy oskarżające kabaret o upadek formy i zubożenie treści. Rozdział czwarty (Kabaret na tle nowych mediów) dotyczy wpływu najnowszych mediów na rozwój kabaretu, a w szczególności zmian, jakie one niosą dla wspólnotowego charakteru kabaretu. W rozdziale piątym (Kabaret jako wspólnota) pojawia się zagadnienie śmiechu grupowego, wspólnoty śmiechu, jaką tworzą artyści i publiczność, a także komunikacyjnego aspektu kabaretu. Wspólnota jest tym, co w kabarecie najważniejsze i konieczne do prawdziwego przeżywania śmiechu. W obecnym świecie najwięcej zagrożeń dla takiego charakteru kabaretu niosą zmiany związane z zapośredniczonym przekazem medialnym, ponieważ niszczą możliwość interakcji między artystami na scenie a widzami.The work is regarding the anthropological dimension of the cabaret as a community. In the first chapter (‘Phenomenon of the cabaret’) the cabaret is characteristic on account of features singling him out of all fields of art which are deciding on the fact that he can be called the art of the laughter. Chapter two (‘Cabaret in Poland’) is presenting the most important cabaret performaces in Poland, including their impact on development of this art. In the chapter three (‘Mechanisms shaping the contemporary Polish cabaret’) brought up matters which considerably influence the appearance of the contemporary cabaret in Poland are remaining as well as voices accusing the cabaret of the fall of the form and impoverishing contents appear. The fourth chapter (‘Cabaret relating to new media’) is regarding the influence of new media on the course of the cabaret performace, in particular changes they are carrying which for the Community character of the cabaret. In the fifth chapter (‘Cabaret as a community’) an issue of the group laughter, community of the laughter artists and an audience form which, as well as the communications aspect of the cabaret appears. Community is it, what in the cabaret most important and necessary for real surviving the laughter. In current world changes connected with media transmission are bringing the most threats to such character of the cabaret, since are destroying the possibility of interaction between artists on the stage and with spectators

Folk love songs from Podegrodzie

Praca dotyczy ludowych pieśni miłosnych, które pochodzą z Podegrodzia - centrum kultury lachowskiej. Teren ten charakteryzuje się dużą liczbą zespołów ludowych, kultywujących wieloletnie tradycje obrzędowo-taneczne, co umożliwiło stosunkowo łatwy dostęp do repertuaru pieśniowego. Dodatkowym walorem zebranych pieśni są pojawiające się w nich cechy gwarowe oraz leksyka typowa dla Podegrodzia. Praca skupia się na ludowych pieśniach miłosnych, w których znaleźć można bogactwo sposobów mówienia o miłości ludowej. Jednym z ciekawszych tematów pieśni miłosnych są zaloty kawalerów do dziewcząt oraz mnogość ich określeń. Analiza objęła również słowa pieszczotliwe pojawiające się w pieśniach podegrodzkich oraz skargi miłosne. Natomiast wizerunek kobiety i mężczyzny, jaki ukazuje się w pieśniach, pozwala na dokładne przyjrzenie się postaciom pieśniowych kochanków. Najważniejszym portretem mężczyzny jest podegrodzianin, który sam siebie ocenia i charakteryzuje w pieśniach i przyśpiewkach. Kobiety są raczej ukazywane poprzez męski pryzmat, a jeśli już wypowiadają się same o sobie, zauważyć można, że ich zachowania w dużej mierze polegają na stosunku do mężczyzny. Interesującym wydaje się pogląd na wartość kandydata na męża ze względu na wiek i stan materialny. Starość w tym wypadku jest zdecydowanie niepożądana w małżeństwie, zwłaszcza z młodą dziewczyną. Również ubóstwo staje się przeszkodą w miłości, przede wszystkim dla mężczyzny. Obrazu miłości ukazanego w pieśniach nie sposób traktować jako całkowicie poważnego wykładnika wiedzy ludu o miłości, jednak niewątpliwie jest on jednym z ciekawszych ujęć tego uczucia w kulturze ludowej.The paper concerns the folk love songs that come from Podegrodzie. This area is characterized by a large number of folk bands, cultivating long-standing tradition of ritual and dance, which allowed relatively easy access to the song’s repertoire. An additional advantage of the collected songs are appearing in their dialect and lexicon features typical of Podegrodzie. The work focuses on the folk songs of love, where you can find a wealth of ways to talk about folk love. One of the interesting themes of love songs are courting bachelors for girls, and the multitude of their terms. The analysis also included words of endearment appearing in songs and complaints about love. In contrast, the image of a man and woman, which appears in the songs, it allows for a close look at the lover’s characters. The most important is the portrait of a man who comes from Podegrodzie. He usually talks well about himself. Women tend to be portrayed by a male prism, and if they speak about themselves, it can be observed that their behaviour largely depend on the ratio of the man. There is an interesting view on the value of a candidate for a husband due to the age and condition of the material. Old age in this case is definitely undesirable in marriage, especially with a young girl. Also, poverty is an obstacle to love, especially for men. Image of love in songs can not be regarded as completely serious exponent of knowledge of the people about love, but it is undoubtedly one of the most interesting shots that feeling in folk culture

Współczesne metody stosowane w analizie biodeterioracji obiektów zabytkowych

Interpretacja zjawiska biodeterioracji obiektów muzealnych wymaga podjęcia działań obejmujących identyfikację dominujących gatunków mikroorganizmów oraz ustalenie związku pomiędzy cechami metabolicznymi zidentyfikowanych mikroorganizmów a właściwościami chemicznymi badanych materiałów. Z tego powodu konieczność opracowywania i wdrażania nowych metod badawczych wydaje się nieodzowna. Do precyzyjnego określenia przynależności taksonomicznej drobnoustrojów pochodzących z badanej próby, również tych niehodowalnych w warunkach laboratoryjnych, przyczynił się rozwój metod molekularnych: genetycznego fingerprintingu (m.in. DGGE/TGGE, T-RFLP, SSCP, ARISA, ARDRA), sekwencjonowania rRNA, narzędzi bioinformatycznych, czy metagenomiki. Oznaczenie określonych biomolekuł na powierzchniach zabytkowych, będących następstwem rozwoju drobnoustrojów, oparte jest na stosunkowo młodej dyscyplinie, metabolomice. Całość dopełniają metody analityczne (takie jak np. SEM, EDX, XRD, FTIR), dzięki którym można określić skutki interakcji drobnoustrojów z materiałem zabytkowym oraz mechanizm biodeterioracji. W artykule przedstawiono i scharakteryzowano współczesne techniki pozwalające ocenić zjawisko biodeterioracji materiałów zabytkowych.Incomplete knowledge of agents responsible for biodeterioration of the museum objects prevents from introducing effective restoration strategies. For proper conservation approaches it is necessary to identify complete microbial consortium inhabiting a given object, as well as, to find connections between products of microbial metabolism and chemical features of the material the object is made of. Therefore, development and application new methods for the study of biodeterioration of historical objects seems to be indispensable. To precisely determine the taxonomic position of microorganisms inhabiting museum objects, the evaluation of different existing molecular techniques is necessary, for instance genetic fingerprinting, ribosomal RNA gene sequencing, bioinformatics or metagenomics. Identification of microbial metabolic products is possible with the help of recently emerged approach, metabolomics. In this paper some current research techniques in use for the evaluation of biodeterioration of historical objects are presented

Secreted form of immunoglobulin M (SIgM)

Immunoglobuliny klasy M to pentameryczne białka określane jako tzw. "immunoglobuliny pierwszego rzutu" produkowane przez limfocyty B. Wśród IgM wyróżnia się formę sekrecyjną (SIgM), która z kolei dzieli się na naturalne i odpornościowe SIgM. Sekrecyjna immunoglobulina klasy M występuje głównie we krwi i bierze udział w utrzymaniu homeostazy limfocytów B, w zakażeniach bakteryjnych, wirusowych, grzybiczych i pasożytniczych, a także w procesach zapalnych, chorobach autoimmunologicznych i miażdżycy. SIgM wiąże antygeny, aktywuje proces fagocytozy i cytotoksyczności komórkowej zależnej od przeciwciał, stanowi także ważny element odporności nabytej, gdyż jako pierwsza pojawia się w odpowiedzi na obecność antygenu. Ponadto, SIgM wspomaga nakierowywanie ciałek apoptotycznych na makrofagi poprzez rekrutacje składnika C1q dopełniacza, usprawniając proces eliminacji produktów apoptozy. Należy dodać, że przeprowadzone do tej pory badania nad tą klasą immunoglobulin wskazują, iż obecność SIgM, szczególnie naturalnej, nie zawsze przynosi pozytywny skutek dla makroorganizmu. Zaobserwowano to w przypadku chorób autoimmunizacyjnych i stanów zapalnych. Jest to jednak ważne i wielofunkcyjne białko mające zdolność do wiązania się z infekującymi organizm drobnoustrojami.Immunoglobulins of class M are pentameric proteins produced by B lymphocytes. Among the IgM's a secretory form (SIgM) is distinguished, which in turn may occur in natural and immune SIgM forms. Secretory immunoglobulin M class occurs mainly in the blood and is involved in B cell homeostasis, bacterial, viral, fungal and parasitic infections, as well as in inflammatory processes, autoimmune diseases, and atherosclerosis. SIgM binds antigens, activates the process of phagocytosis and antibody-dependent cell cytotoxicity. It is also an important component of acquired immunity, because it appears as the first in response to an antigen. In addition, SIgM supports the guiding apoptotic cells by macrophages recruitment of complement component C1q to streamline the process of elimination of apoptotic cells. It should be noted that research carried out so far on this class of immunoglobulins indicates that the presence of SIgM, especially in the natural form, does not always produce a positive effect on macroorganizm. This was observed in the case of autoimmune diseases and inflammatory conditions. However, it is the important and multifunctional protein having the ability to bind to a microbial cells infecting organism

Halophilic microorganisms in deteriorated historic buildings: insights into their characteristics

Historic buildings are constantly being exposed to numerous climatic changes such as damp and rainwater. Water migration into and out of the material's pores can lead to salt precipitation and the so-called efflorescence. The structure of the material may be seriously threatened by salt crystallization. A huge pressure is produced when salt hydrates occupy larger spaces, which leads at the end to cracking, detachment and material loss. Halophilic microorganisms have the ability to adapt to high salinity because of the mechanisms of inorganic salt (KCl or NaCl) accumulation in their cells at concentrations isotonic to the environment, or compatible solutes uptake or synthesis. In this study, we focused our attention on the determination of optimal growth conditions of halophilic microorganisms isolated from historical buildings in terms of salinity, pH and temperature ranges, as well as biochemical properties and antagonistic abilities. Halophilic microorganisms studied in this paper could be categorized as a halotolerant group, as they grow in the absence of NaCl, as well as tolerate higher salt concentrations (Staphylococcus succinus, Virgibacillus halodenitrificans). Halophilic microorganisms have been also observed (Halobacillus styriensis, H. hunanensis, H. naozhouensis, H. litoralis, Marinococcus halophilus and yeast Sterigmatomyces halophilus). With respect to their physiological characteristics, cultivation at a temperature of 25-30°C, pH 6-7, NaCl concentration for halotolerant and halophilic microorganisms, 0-10% and 15-30%, respectively, provides the most convenient conditions. Halophiles described in this study displayed lipolytic, glycolytic and proteolytic activities. Staphylococcus succinus and Marinococcus halophilus showed strong antagonistic potential towards bacteria from the Bacillus genus, while Halobacillus litoralis displayed an inhibiting ability against other halophiles

Coronins and their role in immunological phenomena

Coronins are a large family of proteins occurring in many eukaryotes. In mammals, seven coronin genes have been identified, evidencing that coronins 1 to 6 present classic coronin structure, while coronin 7 is a tandem coronin particle, without a spiral domain, although the best characterised coronin, in terms of both structure and function, is the mammalian coronin 1. It has been proven that they are related to regulation of actin dynamics, e.g. as a result of interaction with the complex of proteins Arp2/3. These proteins also modulate the activity of immune system cells, including lymphocyte T and B cells, neutrophils and macrophages. They are involved in bacterial infections with Mycobacterium tuberculosis, M. leprae and Helicobacter pylori and participate in the response to viral infections, e.g. infections of lymphocytic choriomeningitis virus (LCMV) and vesicular stomatitis Indiana virus (VSV). Also their involvement in autoimmune diseases such as lupus erythematosus has been recorded

Herpes simplex virus type 2 glycoprotein G is targeted by the Sulfated oligo- and polysaccharide inhibitors of virus attachment to cells

Variants of herpes simplex virus type 2 (HSV-2) generated by virus passage in GMK-AH1 cells in the presence of the sulfated oligosaccharide PI-88 were analyzed. Many of these variants were substantially resistant to PI-88 in their initial infection of cells and/or their cell-to-cell spread. The major alteration detected in all variants resistant to PI-88 in the initial infection of cells was a frameshift mutation(s) in the glycoprotein G (gG) gene that resulted in the lack of protein expression. Molecular transfer of the altered gG gene into the wild-type background confirmed that the gG-deficient recombinants were resistant to PI-88. In addition to PI-88, all gG-deficient variants of HSV-2 were resistant to the sulfated polysaccharide heparin. The gG-deficient virions were capable of attaching to cells, and this activity was relatively resistant to PI-88. In addition to having a drug-resistant phenotype, the gG-deficient variants were inefficiently released from infected cells. Purified gG bound to heparin and showed the cell-binding activity which was inhibited by PI-88. Many PI-88 variants produced syncytia in cultured cells and contained alterations in gB, including the syncytium-inducing L792P amino acid substitution. Although this phenotype can enhance the lateral spread of HSV in cells, it conferred no virus resistance to PI-88. Some PI-88 variants also contained occasional alterations in gC, gD, gE, gK, and UL24. In conclusion, we found that glycoprotein gG, a mucin-like component of the HSV-2 envelope, was targeted by sulfated oligo- and polysaccharides. This is a novel finding that suggests the involvement of HSV-2 gG in interactions with sulfated polysaccharides, including cell surface glycosaminoglycans