Liberomyces gen. nov. with two new species of endophytic coelomycetes from broadleaf trees

During a study of endophytic and saprotrophic fungi in the sapwood and phloem of broadleaf trees (Salix alba, Quercus robur, Ulmus laevis, Alnus glutinosa, Betula pendula) fungi belonging to an anamorphic coelomycetous genus not attributable to a described taxon were detected and isolated in pure culture. The new genus, Liberomyces, with two species, L. saliciphilus and L. macrosporus, is described. Both species have subglobose conidiomata containing holoblastic sympodial conidiogenous cells. The conidiomata dehisce irregularly or by ostiole and secrete a slimy suspension of conidia. The conidia are hyaline, narrowly allantoid with a typically curved distal end. In L. macrosporus simultaneous production of synanamorph with thin filamentous conidia was observed occasionally. The genus has no known teleomorph. Related sequences in the public databases belong to endophytes of angiosperms. Phylogenetic analysis revealed a position close to the Xylariales (Sordariomycetes), but family and order affiliation remained unclear.     

Changes in ribosephosphate isomerase and ribosephosphate pyrophosphokinase activities in tobacco infected with PVY

Changes in ribosephosphate isomerase and ribosephosphate pyrophosphokinase activities occurring in tobacco leaf tissues infected with the potato virus Y (PVY) were studied at the stage of acute infection. The results obtained have shown that during the entire experimental period the activities of both enzymes were at the end of a dark phase much higher in virus-infected tissues compared with the values found in healthy control plants. The courses of the activity curves of both enzymes were consistent and correlated with the reproduction curve of PVY. The results obtained suggest a direct involvement of both enzymes inde novo biosynthesis of the virus RNAvia the oxidative pentose phosphate pathway.     

New data on the cytology of parthenogenetic weevils (Coleoptera,Curculionidae)

Parthenogenesis and, in particular, polyploidy are rare in animals. A number of cases, known among weevils, represent apomictic parthenogenesis-a reproductive mode in which eggs undergo one maturation division, the chromosomes divide equationally, and no reduction takes place. Among parthenogenetic weevils there are two diploids, 48 triploids, 18 tetraploids, six pentaploids, three hexaploids and one decaploid. Eight examined parthenogenetic species are triploids with 33 chromosomes of different morphology, confirming that triploidy is the most common level of ploidy in weevils. The karyotypes are heterogeneous with the presence of meta-, submeta-, subtelo- and acrocentric chromosomes. The C-banding method showed that only two species possess a large amount of heterochromatin visible as a band around the centromere during mitotic metaphase. This agrees with observations that weevils are characterized by a small amount of heterochromatin, undetectable in metaphase plates after C-banding. In three species an atypical course of apomictic oogenesis occurs with stages similar to meiosis, in which chromosomes form bivalents and multivalent clusters. This association of chromosomes probably represents the remnants of meiosis, although these events have nothing to do with recombination. The results support the hypothesis that the evolution of apomictic parthenogenesis in weevils has proceeded through a stage of automixis.     

QF-PCR examination of parental and meiotic origin of trisomy 21 in Central and Eastern Europe.

Study of parental/meiotic origin of free trisomy 21 in nuclear families from Russia (70 cases), Ukraine (32 cases), and 22 from Germany revealed maternal nondisjunction in 77.3% (Germany), 93.8% (Ukraine), and 91.4% (Russia), paternal origin in 13.6%, 6.2%, and 8.6%, respectively. Maternal meiosis I errors were found in 84.4% (Ukraine), 77.1% (Russia), paternal origin in 3.1% (Ukraine), 2.9% (Russia). Maternal meiosis II errors occurred in 9.4% and 14.3% and paternal in 3.1% and 5.7% in Ukraine and Russia, respectively. No significant differences were found in maternal/paternal origin among Ukraine, Russia, Germany, and published data from other European regions.     

The value of dynamic contrast enhanced breast MRI in mammographically detected BI-RADS 5 microcalcifications

Aims: To evaluate the role of dynamic contrast-enhanced magnetic resonance imaging (MRI) in the diagnosis of BI-RADS-5 type of microcalcifications of the breast, to compare the size of the microcalcification lesions using mammography (MG) and MRI, and to determine the value of MRI in surgery for microcalcifications. The study also determines the morphology of microcalcification lesions, assesses kinetic curves and compare MRI features of ductal carcinoma in situ (DCIS) for different histopathological grades. Methods: Our group consisted of 32 patients with mammographically detected BI-RADS 5 microcalcifications. The MRI was done in this group of women which was later followed by stereotactic vaccum-assisted biopsy (SVAB). Surgery was performed on all patients with a biopsy that resulted in a diagnosis of breast cancer or atypical ductal hyperplasia (ADH). Results: Of our group of 32 patients, there were 35 mammograhically detected microcalcification lesions, 32 DCIS, one ADH and two benign findings according to the final histology. The microcalcification lesions were larger using MRI than in MG in 10 women. We diagnosed DCIS multifocality in 6 women and bilateral carcinoma in one woman. As with kinetic curve assessment, we found in 67 % of DCIS a rapid rise, 27 % a moderate and in 6 % a slow initial rise. With the pattern of enhancement in the delayed phase, we found in 30 % of DCIS a washout pattern, 67 % a plateau and in 3 % a persistent pattern. Noted difference between high and low grade DCIS was confirmed. Conclusions: MRI sensitivity in the detection of DCIS was 94 % in our group of patients and was the sole evidence for detection of multifocality and bilateral incidence of carcinoma. In 26 % of women the outcome of MRI was the most important for converting breast conserving surgery to mastectomy.     

Accumulation, activity and localization of cell cycle regulatory proteins and the chloroplast division protein FtsZ in the alga Scenedesmus quadricauda under inhibition of nuclear DNA replication

Synchronized cultures of the green alga Scenedesmus quadricauda were grown in the absence (untreated cultures) or in the presence (FdUrd-treated cultures) of 5-fluorodeoxyuridine, the specific inhibitor of nuclear DNA replication. The attainment of commitment points, at which the cells become committed to nuclear DNA replication, mitosis and cellular division, and the course of committed processes themselves were determined for cell cycle characterization. FdUrd-treated cultures showed nearly unaffected growth and attainment of the commitment points, while DNA replication(s), nuclear division(s) and protoplast fission(s) were blocked. Interestingly, the FdUrd-treated cells possessed a very high mitotic histone H1 kinase activity in the absence of any nuclear division(s). Compared with the untreated cultures, the kinase activity as well as mitotic cyclin B accumulation increased continuously to high values without any oscillation. Division of chloroplasts was not blocked but occurred delayed and over a longer time span than in the untreated culture. The FtsZ protein level in the FdUrd-treated culture did not exceed the level in the untreated culture, but rather, in contrast to the untreated culture, remained elevated. FtsZ structures were both localized around pyrenoids and spread inside of the chloroplast in the form of spots and mini-rings. The abundance and localization of the FtsZ protein were comparable in untreated and FdUrd-treated cells until the end of the untreated cell cycle. However, in the inhibitor-treated culture, the signal did not decrease and was localized in intense spots surrounding the chloroplast/cell perimeter; this was in agreement with both the elevated protein level and persisting chloroplast division.     

Treatment with HPMA copolymer-based doxorubicin conjugate containing human immunoglobulin induces long-lasting systemic anti-tumour immunity in mice

Linkage of doxorubicin (Dox) to a water-soluble synthetic N-(2-hydroxypropyl)methacrylamide copolymer (PHPMA) eliminates most of the systemic toxicity of the free drug. In EL-4 lymphoma-bearing C57BL/6 mice, a complete regression of pre-established tumours has been achieved upon treatment with Dox–PHPMA–HuIg conjugate. The treatment was effective using a range of regimens and dosages, ranging from 62.5 to 100% cured mice treated with a single dose of 10–20 mg of Dox eq./kg, respectively. Fractionated dosages producing lower levels of the conjugate for a prolonged time period had substantial curative capacity as well. The cured mice developed anti-tumour protection as they rejected subsequently re-transplanted original tumour. The proportion of tumour-protected mice inversely reflected the effectiveness of the primary treatment. The treatment protocol leading to 50% of cured mice produced only protected mice, while no mice treated with early treatment regimen (i.e. starting on day 1 after tumour transplantation) rejected the re-transplanted tumour. Exposure of the host to the cancer cells was a prerequisite for developing protection. The anti-tumour memory was long lasting and specific against the original tumour, as the cured mice did not reject another syngeneic tumour, melanoma B16-F10. The immunity was transferable to naïve recipients in in vivo neutralization assay by spleen cells or CD8⁺ lymphocytes derived from cured animals. We propose an effective treatment strategy which eradicates tumours without harming the protective immune anti-cancer responses.