The biotechnology of higher fungi - current state and perspectives

This review article concisely describes methodology of biotechnological processes with the use of cultures of higher fungi, their application in bioremediation and to obtain biologically active preparations. Advantages and disadvantages of biotechnological methods used to cultivate mushrooms are analyzed. This paper contains overview of higher fungi species most commonly used in biotechnological processes, of cultivation methods applied to produce fungal biomass, of enzymes and bioactive metabolites and of the strategies for submerged cultivation of the mycelial cultures. The problems of optimization of strains and biotechnological processes are briefly discussed.Niemniej pomimo trudności, producenci substancji leczniczych pochodzenia grzybowego (Lentinan, LEM, Grifon-D, PSK, PSP), suplementów diety oraz enzymów grzybowych, wprowadzają metody biotechnologiczne do produkcji. Zgodnie ze stosowanym od dawna w biotechnologii przemysłowej (np. przez producentów antybiotyków) zwyczajem, warunki procesu rzadko są opisywane w publikacjach, a czasami nie są nawet patentowane - co ułatwia zachowanie ich w tajemnicy. W latach 90-tych XX wieku pojawiły się pierwsze informacje o możliwości stosowania metod rekombinowanego DNA dla grzybów wyższych. Współcześnie, liczne publikacje donoszą o opracowaniu metod transformacji oraz o uzyskaniu modyfikowanych genetycznie grzybów jadalnyc

Identification of bacteria and fungi inhabiting fruiting bodies of Burgundy truffle (Tuber aestivum Vittad.)

Tuber species may be regarded as complex microhabitats hosting diverse microorganisms inside their fruiting bodies. Here, we investigated the structure of microbial communities inhabiting the gleba of wild growing (in stands) T. aestivum, using Illumina sequencing and culture-based methods. The two methods used in combination allowed to extract more information on complex microbiota of Tuber aestivum gleba. Analysis of the V3–V4 region of 16S rDNA identified nine phyla of bacteria present in the gleba of T. aestivum ascomata, mostly Proteobacteria from the family Bradyrhizobiaceae. Our results ideally match the earlier data for other Tuber species where the family Bradyrhizobiaceae was the most represented. The ITS1 region of fungal rDNA represented six alien fungal species belonging to three phyla. To complement the metagenomic analysis, cultivable fungi and bacteria were obtained from the gleba of the same T. aestivum fruiting bodies. The identified fungi mostly belong to the phylum Basidiomycota and same to Ascomycota. Analysis of cultivable bacteria revealed that all the specimens were colonized by different strains of Bacillus. Fungal community inhabiting T. aestivum fruiting bodies was never shown before

Synthesis of new 4-butyl-arylpiperazine-3-(1H-indol-3-yl)pyrrolidine-2,5-dione derivatives and evaluation for their 5-HT1A_{1A} and D2_{2} receptor affinity and serotonin transporter inhibition

A series of novel 4-butyl-arylpiperazine-3-(1H-indol-3-yl)pyrrolidine-2,5-dione derivatives were synthesized and evaluated for their 5-HT$_{1A}$/D2 receptor affinity and serotonin reuptake inhibition. The compounds exhibited high affinity for the 5-HT$_{1A}$ receptor, (especially 4d K$_{i}$ = 0.4 nM) which depended on the substitution pattern at the phenylpiperazine moiety. From this series screen, compound 4c emerged with promising mixed receptor profiles for the 5-HT$_{1A}$/D2 receptors and the serotonin transporter (K$_{i}$ = 1.3 nM, 182 nM and 64 nM, respectively).long-chain arylpiperazinesdepression5-HT$_{1A}$ receptor ligandsschizophreniamulti-target ligand

Selenium-Containing Polysaccharides—Structural Diversity, Biosynthesis, Chemical Modifications and Biological Activity

Selenosugars are a group of sugar derivatives of great structural diversity (e.g., molar masses, selenium oxidation state, and selenium binding), obtained as a result of biosynthesis, chemical modification of natural compounds, or chemical synthesis. Seleno-monosaccharides and disaccharides are known to be non-toxic products of the natural metabolism of selenium compounds in mammals. In the case of the selenium-containing polysaccharides of natural origin, their formation is also postulated as a form of detoxification of excess selenium in microorganisms, mushroom, and plants. The valency of selenium in selenium-containing polysaccharides can be: 0 (encapsulated nano-selenium), IV (selenites of polysaccharides), or II (selenoglycosides or selenium built into the sugar ring to replace oxygen). The great interest in Se-polysaccharides results from the expected synergy between selenium and polysaccharides. Several plant- and mushroom-derived polysaccharides are potent macromolecules with antitumor, immunomodulatory, antioxidant, and other biological properties. Selenium, a trace element of fundamental importance to human health, has been shown to possess several analogous functions. The mechanism by which selenium exerts anticancer and immunomodulatory activity differs from that of polysaccharide fractions, but a similar pharmacological effect suggests a possible synergy of these two agents. Various functions of Se-polysaccharides have been explored, including antitumor, immune-enhancement, antioxidant, antidiabetic, anti-inflammatory, hepatoprotective, and neuroprotective activities. Due to being non-toxic or much less toxic than inorganic selenium compounds, Se-polysaccharides are potential dietary supplements that could be used, e.g., in chemoprevention

Synthesis of New Perhydropyrrolo[1,2-a]pyrazine Derivatives and Their Evaluation in Animal Models of Epilepsy

A series of novel stereochemically pure derivatives of the investigative broad-spectrum anticonvulsant ADD408003 was designed and synthesized. Five-center four-component (U-5C-4CR) and four-center three-component (U-4C-3CR) variants of Ugi reaction were used in the key step of the synthetic pathways. The compounds obtained were evaluated for the anticonvulsant activitiy in the maximal electroshock seizure (MES), subcutaneous Metrazole (scMET) and minimal clonic seizure (6 Hz) animal models of epilepsy. The efficacies of most derivatives in the 6 Hz model of pharmacoresistant partial seizures were markedly higher than in the ‘classical’ MES and scMET models. The most active compounds, (4R,8aR)-3a, and (4S,8aS)-6 displayed median effective doses (ED50) of 47.90 and 126.19 mg/kg, respectively, for the 6 Hz test

Stabilność chemiczna klejów ortodontycznych opartych na sieci polimerowej w zależności od pH środowiska zewnętrznego

Chemical stability of composite adhesive systems is crucial for the safety of their use.The study assessed chemical stability of four light-cured orthodontic adhesives: Contec LC, Transbond XT, Transbond Plus, Resilience, depending on pH value of the external environment. Samples of polymerized orthodontic adhesives were treated with (high-performance liquid chromatography) HPLC-grade water solutions of phosphate-citrate buffer with pH values respectively: 4, 5,6 and 7 at 36 °C. The eluates obtained after 1 hour, 24 hours and 7 days of sample incubation were analyzed for the presence of camphorquinone (CQ), bisphenol A (BPA), triethylene glycol dimethacrylate (TEGDMA), urethane dimethacrylate (UDMA), bisphenol A diglycidyl methacrylate (Bis-GMA), ethylene glycol dimethacrylate (EGDMA), 2,2-dimethoxy-2-phenylacetophenon (DMPA) usingultra-high performance liquid chromatography (UHPLC). Out of the seven searchable substances, TEGDMA was present in eluates obtained from Contec LC, Resilience and Transbond XT materials and EGDMA in eluates obtained from Resilience adhesive. The eluates obtained from the Transbond Plus adhesive system were virtually free of the sought substances. The highest concentrations of TEGDMA in solutions were recorded after 1 hour of incubation regardless ofthe type of material. In the case of Contec LC material, an increase in TEGDMA concentrations was observed along with an increase in the solutions’ pH, butonly for the elution period of 1 hour and 7 days, the effect of the solvent’s pH was statistically significant (p ≤ 0.001). In the case of Resilience and Transbond XT, no significant differences in TEGDMA concentrations were observed with respect to pH of the external environment. In the conditions of the conducted study, a lack of chemical stability was confirmed for the majority of assessed orthodontic adhesive systems based on polymers, expressed in emission of component monomers to the external environment. The chemical compound identified in the study was TEGDMA, and for each pH of the solvent, statistically significant differences in its release were found between the materials. However, no explicit relationship was observed between chemical in stability of the studied materials and pH of the external environment within the assumed range of assessment.Stabilność chemiczna kompozytowych systemów adhezyjnych jest kluczowa z punktu widzenia bezpieczeństwa ich stosowania. W badaniu oceniano stabilność chemiczną czterech światłoutwardzalnych klejów ortodontycznych: Contec LC, Transbond XT, TransbondPlus, Resilience, w zależności od wartości pH środowiska zewnętrznego. Próbki spolimeryzowanych klejów ortodontycznych poddano działaniu roztworów buforufosforanowo-cytrynianowego na bazie wody o czystości HPLC, o wartości pH: 4, 5,6 oraz 7 i temperaturze 36 °C. Eluaty uzyskane po 1 h, 24 h i 7 dniach inkubacji próbek analizowano metodą chromatografii cieczowej wysokociśnieniowej (HPLC) pod względem obecności kamforochinonu (CQ), bisfenolu A (BPA), dimetakrylanu glikolu trietylenowego (TEGDMA), dimetakrylanu uretanu (UDMA), bisfenolu A metakrylanu diglicydylu (Bis-GMA), dimetakrylanu glikoluetylenowego (EGDMA), 2,2-dimetoksy-2-fenyloacetofenonu (DMPA). Z siedmiu związków chemicznych identyfikowanych w roztworach potwierdzono obecność TEGDMA w eluatach uzyskanych z materiałów Contec LC, Resilience i Transbond XT oraz obecność EGDMA w eluatach z kleju Resilience. Eluaty otrzymane z systemu adhezyjnego TransbondPlus praktycznie biorąc nie zawierały poszukiwanych substancji. Największe stężenia TEGDMA w roztworach stwierdzono po 1 h inkubacji próbek ortodontycznych systemów łączących, niezależnie od rodzaju materiału. W odniesieniu do kleju Contec LC obserwowano wzrost stężenia TEGDMA wraz zwartością pH roztworów, ale wpływ pH rozpuszczalnika był istotny statystycznie (p ≤ 0,001) tylko w wypadku czasu wymywania 1 h i 7 dni. W roztworach po inkubacji materiałów Resilience i Transbond XT nie stwierdzono istotnych różnic stężeń TEGDMA w zależności od pH środowiska zewnętrznego. W warunkach przeprowadzonego badania potwierdzono brak stabilności chemicznej większości ocenianych, polimerowych, ortodontycznych systemów adhezyjnych wyrażający się emisją tworzących je monomerów do środowiska zewnętrznego. W odniesieniu do każdej wartości pH rozpuszczalnika wykazano istotne statystycznie różnice w uwalnianiu TEGDMA pomiędzy badanymi materiałami. Jednocześnie w przyjętym zakresie oceny nie zaobserwowano jednoznacznej zależności stabilności chemicznej badanych materiałów od pH środowiska zewnętrznego

Immunomodulatory Properties of Polysaccharides from Lentinula edodes

Lentinula edodes (Berk.) Pegler, also known as shiitake mushroom, is a popular edible macrofungus and a source of numerous bioactive substances with multiple beneficial health effects. L. edodes-derived polysaccharides are the most valuable compounds, with anticancer, antioxidant, antimicrobial, and immunomodulatory properties. It has been demonstrated that their biological activity depends on the extraction method, which affects monosaccharide composition, molecular weight, branching degrees, and helical conformation. In this review, we discuss the immunomodulatory properties of various polysaccharides from L. edodes in animal models and in humans