Are cysteine and its derivatives applicable for the prevention of fungal diseases?

Many filamentous fungi are destructive pathogens of plants and are thus responsible for enormous crop losses worldwide. Therefore, there is a substantial demand for safely applicable, new compounds with extensive antifungal activity. Cysteine and its derivatives are interesting from this respect, as they have effective inhibitory potential against microorganisms and do not harmful effect on animal and plant cells. Cysteine is a non-essential amino acid, which is an important structural and functional component of many proteins and enzymes. Previous studies demonstrated that cysteine and N-acetyl-cysteine have also been shown to exert antifungal activity. A number of members of the class Zygomycetes are important as postharvest pathogens of agricultural products; Rhizopus, Mucor and Gilbertella species are among the most frequently isolated causative agents of fungal rots. In this study we investigated the in vitro antifungal activity of cysteine (D- and L-cysteine) and its 4 derivatives (L-cysteine-methyl-ester, N-acetyl-cysteine, N-isobutyryl-D-cysteine and N-isobutyryl-L-cysteine) against 20 zygomyceteous fungal isolates representing 16 genera (Actinomucor, Backusella, Gilbertella, Lichtheimia, Micromucor, Mortierella, Mucor, Mycotypha, Phycomyces, Rhizomucor, Rhizopus, Saksenaea, Syncephalastrum, Thamnostylum, Umbellopsis and Zygorynchus). The inhibitory potential of different concentrations (ranging from 0.625 to 10 mM) of the cysteine and its derivatives were investigated on the germination of sporangiospores and on hyphal extension by broth microdilution method and agar plate test. Cysteine and its derivatives showed a strong inhibitory effect against the most studied strains. Treatment with 10 mM of compounds resulted total growth inhibition in case of some isolates. Severe changes in colony morphology and hyphal growth were observed in presence of 10 mM Lcysteine, N-acetyl-cysteine and N-isobutyryl-L-cysteine when a strain was sensitive to them. Ten mM N-acetyl-cysteine induced dramatic modifications in the structural organization of the hyphae in case of Rhizopus stolonifer. The above mentioned features of the investigated compounds could make them favourable antifungal agents against Zygomycetes in agricultural respect in the future, but it is needed further studies to prove their practical efficiency, for example in plant model experiments

A hőkezelés alkalmazási lehetőségei a kriminalisztikai üvegvizsgálatokban

Cél: Az alábbiakban célunk bemutatni a Nemzeti Szakértői és Kutató Központban végzett, üvegek hőkezelésével kapcsolatos kísérletek eredményeit, valamint a hőkezelés alkalmazási lehetőségeit a kriminalisztikai vizsgálatokban. Módszertan: Az üvegek törésével is együtt járó bűncselekményekkel (például gépjárműfeltörés, betöréses lopás, rongálás) kapcsolatosan jellemzően felmerülő, szakértői vizsgálatot igénylő kérdés, hogy a lefoglalt ruházati tárgyon talált üvegszemcse származhat-e a bűncselekmény helyszínén eltört üvegtárgyak valamelyikéből. Mikroméretű üvegszemcsék kriminalisztikai vizsgálatának elterjedt módszere az optikai törésmutató mérés. Az üvegmaradvány-szemcse és a referenciaminta törésmutatójának azonossága esetén valószínűsíthető a származási kapcsolat. Az ilyen jellegű vizsgálatok kiegészítéseként szolgál az üvegszemcsék kontrollált körülmények között történő hőkezelése és az azt követő optikai törésmutató mérés. Jelen munkánk során ezt a vizsgálati lehetőséget mutatjuk be példákon keresztül, az eredményeket összevetve korábbi irodalmi adatokkal. Megállapítások: Hőkezelés hatására az üvegszemcsén belül szerkezeti átrendeződés történik, melynek hatására a szemcse optikai törésmutatója megváltozik. Ennek mértéke a különböző típusú üvegekben jelen lévő, eltérő belső feszültség miatt üvegtípusonként különböző és jellegzetes. Ez lehetőséget nyújt a pontosabb származási viszony megállapítása mellett az üveg típusának meghatározására is. A hőkezelést olyan esetekben is sikerrel alkalmaztuk, ahol a bűncselekményt követően a referenciamintát ért jelentős hőterhelés (például tűz) miatt az üvegmaradvány-szemcsékkel optikai törésmutató alapján történő közvetlen összehasonlítás téves eredményt adna. Érték: Sikeresen alkalmaztuk a hőkezelést üvegtípus megállapítására, származási viszonyok vizsgálatára, valamint hőterhelésnek kitett üvegek analíziséreis, ezzel segítve a büntetőügyekben felmerülő szakkérdések megválaszolását

A hőkezelés alkalmazási lehetőségei a kriminalisztikai üvegvizsgálatokban

Aim: In this paper, our primary aim is to present the results of our experiments, carried out at the Hungarian Institute for Forensic Sciences, as well as the possible applications of annealing in the forensic investigation of glasses.Methodology: The question that typically arises and requires forensic examinations in connection with crimes involving the breaking of glasses (e. g. burglary, vandalism) is whether the glass fragments recovered from the suspect’s garment can originate from any of the broken samples found on the scene of the crime. A common method for forensic investigation of glass microfragments is the refractive index measurement. In cases when the refractive index of the recovered fragments and the reference is the same, it is probable that they originate from the same source. These investigations can be supplemented by the heat treatment of the glass samples under controlled conditions followed by the measurement of the refractive index. In this work we present this method via examples, comparing the results with previous literature data.Findings: As a result of annealing, a structural rearrangement takes place inside the glass fragments and the refractive index changes. The extent of this change is characteristic of the glass types due to the different internal stress present in the different types. Besides establishing a more precise relationship of origin, this method also makes it possible to determine glasses by type. Annealing has also been successfully applied in cases where direct comparison of the recovered fragments with the reference sample would have given false results due to the significant heat effect (e. g. fire) on the control sample after the crime.Value: We have successfully applied annealing to determine the type of glasses, to examine the possibilities of origin, and to analyze glasses exposed to heat, thus helping to answer forensic questions in criminal cases.Cél: Az alábbiakban célunk bemutatni a Nemzeti Szakértői és Kutató Központban végzett, üvegek hőkezelésével kapcsolatos kísérletek eredményeit, valamint a hőkezelés alkalmazási lehetőségeit a kriminalisztikai vizsgálatokban.Módszertan: Az üvegek törésével is együtt járó bűncselekményekkel (például gépjárműfeltörés, betöréses lopás, rongálás) kapcsolatosan jellemzően felmerülő, szakértői vizsgálatot igénylő kérdés, hogy a lefoglalt ruházati tárgyon talált üvegszemcse származhat-e a bűncselekmény helyszínén eltört üvegtárgyak valamelyikéből. Mikroméretű üvegszemcsék kriminalisztikai vizsgálatának elterjedt módszere az optikai törésmutató mérés. Az üvegmaradvány-szemcse és a referenciaminta törésmutatójának azonossága esetén valószínűsíthető a származási kapcsolat. Az ilyen jellegű vizsgálatok kiegészítéseként szolgál az üvegszemcsék kontrollált körülmények között történő hőkezelése és az azt követő optikai törésmutató mérés. Jelen munkánk során ezt a vizsgálati lehetőséget mutatjuk be példákon keresztül, az eredményeket összevetve korábbi irodalmi adatokkal.Megállapítások: Hőkezelés hatására az üvegszemcsén belül szerkezeti átrendeződés történik, melynek hatására a szemcse optikai törésmutatója megváltozik. Ennek mértéke a különböző típusú üvegekben jelen lévő, eltérő belső feszültség miatt üvegtípusonként különböző és jellegzetes. Ez lehetőséget nyújt a pontosabb származási viszony megállapítása mellett az üveg típusának meghatározására is. A hőkezelést olyan esetekben is sikerrel alkalmaztuk, ahol a bűncselekményt követően a referenciamintát ért jelentős hőterhelés (például tűz) miatt az üvegmaradvány-szemcsékkel optikai törésmutató alapján történő közvetlen összehasonlítás téves eredményt adna.Érték: Sikeresen alkalmaztuk a hőkezelést üvegtípus megállapítására, származási viszonyok vizsgálatára, valamint hőterhelésnek kitett üvegek analíziséreis, ezzel segítve a büntetőügyekben felmerülő szakkérdések megválaszolását

Are cysteine and its derivatives applicable for the prevention of fungal diseases?

Many filamentous fungi are destructive pathogens of plants and are thus responsible for enormous crop losses worldwide. Therefore, there is a substantial demand for safely applicable, new compounds with extensive antifungal activity. Cysteine and its derivatives are interesting from this respect, as they have effective inhibitory potential against microorganisms and do not harmful effect on animal and plant cells. Cysteine is a non-essential amino acid, which is an important structural and functional component of many proteins and enzymes. Previous studies demonstrated that cysteine and N-acetyl-cysteine have also been shown to exert antifungal activity. A number of members of the class Zygomycetes are important as postharvest pathogens of agricultural products; Rhizopus, Mucor and Gilbertella species are among the most frequently isolated causative agents of fungal rots. In this study we investigated the in vitro antifungal activity of cysteine (D- and L-cysteine) and its 4 derivatives (L-cysteine-methyl-ester, N-acetyl-cysteine, N-isobutyryl-D-cysteine and N-isobutyryl-L-cysteine) against 20 zygomyceteous fungal isolates representing 16 genera (Actinomucor, Backusella, Gilbertella, Lichtheimia, Micromucor, Mortierella, Mucor, Mycotypha, Phycomyces, Rhizomucor, Rhizopus, Saksenaea, Syncephalastrum, Thamnostylum, Umbellopsis and Zygorynchus). The inhibitory potential of different concentrations (ranging from 0.625 to 10 mM) of the cysteine and its derivatives were investigated on the germination of sporangiospores and on hyphal extension by broth microdilution method and agar plate test. Cysteine and its derivatives showed a strong inhibitory effect against the most studied strains. Treatment with 10 mM of compounds resulted total growth inhibition in case of some isolates. Severe changes in colony morphology and hyphal growth were observed in presence of 10 mM Lcysteine, N-acetyl-cysteine and N-isobutyryl-L-cysteine when a strain was sensitive to them. Ten mM N-acetyl-cysteine induced dramatic modifications in the structural organization of the hyphae in case of Rhizopus stolonifer. The above mentioned features of the investigated compounds could make them favourable antifungal agents against Zygomycetes in agricultural respect in the future, but it is needed further studies to prove their practical efficiency, for example in plant model experiments

Taxonomic and metabolic investigation of bipolaris species

The ascomycetous genus Bipolaris (Euascomycetes, Pleosporales, Pleosporaceae) contains melanin producing filamentous fungal species. Teleomorphic stages belong to the genus Cochliobolus together with the members of the other anamorphic genus, Curvularia; however, sexual reproduction has been rarely observed in nature. These fungi are frequently associated to graminaceous hosts and may infect corn, wheat, barley and rice causing devastating epidemics from time to time, primarily in tropical and subtropical regions. Climate change favours the spreading of these species in temperate regions, which underlines the importance of their study. The symptoms of Bipolaris infections begin with small necrotic lesions on leafs, which extends and cause notable yield losses. Bipolaris species produce a series of biological active secondary metabolites, which have or may have role in the pathogenesis. They produce various terpenoid compounds, such as sesquiterpenes, diterpenes and sesterterpenes. The sesterterpene-type ophiobolins constitute one of the most remarkable groups of these bioactive metabolites. Apart from their phytotoxic effects, several ophiobolin analogues have antimicrobial, antiviral, cytotoxic, anticancer, or nematocidal activity. Besides plant pathogenicity, the genus is also known about their difficult species identification and confused taxonomy. Traditional morphology based methods are unsuitable for the precise species identification because of the similar characteristics, molecular identification based on the frequently used nuclear ribosomal ITS region is also problematic. In the present study, new markers potentially suitable for molecular identification were tested. Our study demonstrated that the ITS region, the calm, tub and tef gene sequences, which are routinely used for identification, are unsuitable to discern Bipolaris species. Instead of them the use of the intergenic spacer region (IGS) of the nuclear ribosomal RNA gene cluster is suggested, in which species specific motifs were determined to distinguish the Bipolaris isolates. Furthermore, ability of Bipolaris and Cochliobolus isolates representing 23 different species to produce ophiobolin A, the best known phytotoxin of these fungi, was also investigated. Six of the tested isolates produced remarkable amounts of ophiobolin A (>1 mg/g [dry weight]). The secretion kinetics of the examined strains has been determined by HPLC technique during a 12 days long cultivation and found to be aggregated in four different groups according to the results. This study forms part of the project TÁMOP-4.2.2.A-11/1/KONV-2012-0035, supported by the European Union, co-financed by the European Social Fund. For Cs.V: this research was realized in the frames of TÁMOP 4.2.4. A/2-11-1-2012-0001 „National Excellence Program – Elaborating and operating an inland student and researcher personal support system”. The project was subsidized by the European Union and co-financed by the European Social Fund

Genomes of fungi and relatives reveal delayed loss of ancestral gene families and evolution of key fungal traits

Fungi are ecologically important heterotrophs that have radiated into most niches on Earth and fulfil key ecological services. Despite intense interest in their origins, major genomic trends of their evolutionary route from a unicellular opisthokont ancestor to derived multicellular fungi remain poorly known. Here we provide a highly resolved genome-wide catalogue of gene family changes across fungal evolution inferred from the genomes of 123 fungi and relatives. We show that a dominant trend in early fungal evolution has been the gradual shedding of protist genes and the punctuated emergence of innovation by two main gene duplication events. We find that the gene content of non-Dikarya fungi resembles that of unicellular opisthokonts in many respects, owing to the conservation of protist genes in their genomes. The most rapidly duplicating gene groups included extracellular proteins and transcription factors, as well as ones linked to the coordination of nutrient uptake with growth, highlighting the transition to a sessile osmotrophic feeding strategy and subsequent lifestyle evolution as important elements of early fungal history. These results suggest that the genomes of pre-fungal ancestors evolved into the typical filamentous fungal genome by a combination of gradual gene loss, turnover and several large duplication events rather than by abrupt changes. Consequently, the taxonomically defined Fungi represents a genomically non-uniform assemblage of species. Fungi exhibit remarkable morphological and ecological diversity. An analysis of the genomes of 123 fungi and relatives shows gradual loss of protist genes, major gene turnover and duplication leading to the evolution of modern traits of filamentous fungi