Steppe-like grassland as a refuge of the wild edible and medicinal plant species in anthropogenic landscape in northeastern Croatia

Habitat fragmentation and modification have been recognized as the most serious threats to biodiversity. In Croatia, the intensive human activities resulted in the landscape transformation, reduction of grassland areas and loss of species number. Increasingly threatened are the wild edible and medicinal plants which contribute remarkably to the biodiversity of different grassland types. In northeastern Croatia, one of the last remnants of the steppe-like grassland remained preserved in a very complex anthropogenic landscape. We examined the diversity of vascular plants occurring on this small habitat patch. Among the relatively high number of plant taxa (in a total of 177) found during the two years of investigations, most could be used both for human nutrition (28 taxa), and for the treatment of a wide range of diseases (60 taxa), while some taxa have nutritional value for livestock (17 taxa). All 23 recorded taxa with edible and medicinal properties are threatened in Croatia, 21 are threatened across Europe, and two are Red Listed taxa. Our results showed that small and isolated steppe-like grassland represents an important refuge for variety of valuable plant species in intensively used landscapes. Accordingly, such small habitats should be, at least locally, effectively protected.     

Enzymatic activity of basidiomycetes

In mycelial acetone powder of the basidiomyceteOudemansiella mucida, the presence of enzymatic systems of NADH₂-oxidase and-dehydrogenase type and of NAD⁺-dependent alcoholdehydrogenase (using ethanol as substrate) has been detected. NADH₂-oxidase has its optimal pH value in the region of 6,0 and is not too active. NADH₂-dehydrogenase which reduces 2,6-dichlorophenolindophenol is very active, may be eluted from the mycelial preparation with water and its adequate pH is in the region of 6,2. On the other hand, NADH₂-dehydrogenase reducing triphenyltetrazolium chloride is non-eluable with water, its activity depends on the presence of menadione and its adequate pH lies in the region of 7,4.     

Melectin MAPs: the influence of dendrimerization on antimicrobial and hemolytic activity

The recently described antimicrobial peptide melectin (MEP, GFLSILKKVLPKVMAHMK-NH₂) exhibits high antimicrobial activity against Gram-positive and Gram-negative bacteria. Here we describe the synthesis and biological activities of 23 new analogues of MEP. We studied the influence of dimerization and tetramerization (MAP-constructs of MEP) on the antimicrobial and hemolytic activities, as well as the role of Met in positions 14 and 17 of the peptide chain. Oxidation of the Met to Met(O) and Met(O₂) decreases antimicrobial activity of all tested bacteria if the peptide is in the monomeric form, however, only to Staphylococcus aureus if in the form of dimer or tetramer. Dimerization and tetramerization increase the undesirable hemolytic activity of the peptides. Interestingly, substitution of Leu for Val in position 6 leads to the decrease of hemolytic activity. Introduction of the isosteric amino acid Nle into positions 14 or 17 or both leads to slight increase of hemolytic activity under preservation of high antimicrobial activities. Unfortunately, dimerization again leads to an increase of hemolytic activity.     

Microbial production and purification of arachidonic acid

Summary Mortierella alpina S-17 cultivated in shaker flasks in the semisynthetic liquid medium with 3% glucose at 28 °C for 14 days accumulated 2.1 g/l of intracellular lipid rich in arachidonic acid (ARA). The content of ARA was 50 % of the total fatty acids. The simple method for high pure methyl arachidonate preparation was developed.     

Screening of patients at risk for 22q11 deletion

The aim of this study was to determine whether deletion 22q11.2 studies should become apart of a standardized diagnostic workup for selected groups of at risk patients. We prospectively investigated four cohorts of unselected patients referred because of 1) congenital heart defect (CHD), 2) palatal anomalies, 3) hypocalcaemia, 4) dysmorphic features suggestive of del 22q11.2. Fluorescence in situ hybridization analysis revealed deletion 22q11.2 in 9.4% (6/64) patients with CHD. From 18 patients referred because of the hypocalcaemia, six (33.3%) had 22q11.2 deletion. In the group of 31 children with dysmorphic traits, the diagnosis was confirmed in two (6.4%) patients. None of the 58 children with palatal anomalies showed evidence of 22q11.2 deletion. Conclusions: Testing for the 22q11.2 microdeletion can be recommended in all patients with conotruncal heart defects and in patients with hypocalcaemia. It should be also considered in patients presenting only with dysmorphic traits suggestive of del 22q11.2, while screening in patients with cleft palate is not warranted.     

Coordinated activation of AMP‐activated protein kinase,extracellular signal‐regulated kinase,and autophagy regulates phorbol myristate acetate‐induced differentiation of SH‐SY5Y neuroblastoma cells

We explored the interplay between the intracellular energy sensor AMP‐activated protein kinase (AMPK), extracellular signal‐regulated kinase (ERK), and autophagy in phorbol myristate acetate (PMA)‐induced neuronal differentiation of SH‐SY5Y human neuroblastoma cells. PMA‐triggered expression of neuronal markers (dopamine transporter, microtubule‐associated protein 2, β‐tubulin) was associated with an autophagic response, measured by the conversion of microtubule‐associated protein light chain 3 (LC3)‐I to autophagosome‐bound LC3‐II, increase in autophagic flux, and expression of autophagy‐related (Atg) proteins Atg7 and beclin‐1. This coincided with the transient activation of AMPK and sustained activation of ERK. Pharmacological inhibition or RNA interference‐mediated silencing of AMPK suppressed PMA‐induced expression of neuronal markers, as well as ERK activation and autophagy. A selective pharmacological blockade of ERK prevented PMA‐induced neuronal differentiation and autophagy induction without affecting AMPK phosphorylation. Conversely, the inhibition of autophagy downstream of AMPK/ERK, either by pharmacological agents or LC3 knockdown, promoted the expression of neuronal markers, thus indicating a role of autophagy in the suppression of PMA‐induced differentiation of SH‐SY5Y cells. Therefore, PMA‐induced neuronal differentiation of SH‐SY5Y cells depends on a complex interplay between AMPK, ERK, and autophagy, in which the stimulatory effects of AMPK/ERK signaling are counteracted by the coinciding autophagic response.

     

Immobilization and nonspecific adsorption of proteins to pyrogeneous highly dispersed silicon dioxide

Summary A support based on pyrogeneous silicon dioxide of particle size 0.01 to 0.1/um, modified by 3-(amino)propyltriethoxysilane and activated by glutaraldehyde was employed for the immobilization of concanavalin A, immunoglobulins, basic pancreatic trypsin inhibitor, and chymotrypsin. Its binding capacity is comparable with that of porous supports while the biological activity of the proteins immobilized is retained. Nonspecific adsorption of these proteins to the support is low compared to its binding capacity.     

Extracellular oxidative enzyme production and PAH removal in soil by exploratory mycelium of white rot fungi

Selected strains of three species of white rot fungi, Pleurotus ostreatus, Phanerochaete chrysosporium and Trametes versicolor, were grown in sterilized soil from straw inocula. The respective colonization rates and mycelium density values decreased in the above mentioned order. Three- and four-ringed PAHs at 50 ppm inhibited growth of fungi in soil to some extent. The activities of fungal MnP and laccase (units per g dry weight of straw or soil), extracted with 50 mM succinate-lactate buffer (pH 4.5), were 5 to 20-fold higher in straw compared to soil. The enzyme activities per g dry soil in P. ostreatus and T. versicolor were similar, in contrast to P. chrysosporium, where they were extremely low. Compared to the aerated controls, P. ostreatus strains reduced the levels of anthracene, pyrene and phenanthrene by 81–87%, 84–93% and 41–64% within 2 months, respectively. During degradation of anthracene, all P. ostreatus strains accumulated anthraquinone. PAH removal rates in P. chrysosporium and T. versicolor soil cultures were much lower.