The fine structure of the germinal mass, brood cavity and birth canal of the rediae of the monoxenous hemiuroid digenean Bunocotyle progenetica Chabaud & Buttner, 1959.

Bunocotyle progenetica is a hemiuroid digenean whose sexual adults become fully developed and lay their eggs inside the rediae in the molluscan host. In this study, the fine structure of the germinal mass, brood cavity and birth canal in the B. progenetica rediae was examined using transmission electron and confocal microscopy. The large germinal mass attached to the body wall has a cellular composition typical for this organ. The characteristic traits of this germinal mass are weakly developed supporting tissue and the presence of deep lacunae opening into the brood cavity. These lacunae presumably participate in feeding the deeply lying embryos and facilitate their release into the brood cavity. The germinal mass is also characterized by intensive degeneration of cellular elements, which may represent a mechanism controlling the offspring number, limited in this species by the size of the redial brood cavity. The brood-cavity lining consists of flattened cells bearing lamellar projections and is connected anteriorly with the epithelium of the birth canal. The brood-cavity musculature, which is well developed in other hemiuroid digeneans, is significantly reduced in B. progenetica, most likely because their cystophorous cercariae remain inside the rediae, removing the need for muscle contractions pushing them through the brood cavity. The birth canal comprises three regions distinguished by the structure of the lining and muscle arrangement. The comparison of rediae of B. progenetica with parthenitae of other digeneans has shown that the organization of the redial reproductive apparatus in this species may have been influenced by life-cycle modification.

Patterns and processes influencing helminth parasites of Arctic coastal communities during climate change.

This review analyses the scarce available data on biodiversity and transmission of helminths in Arctic coastal ecosystems and the potential impact of climate changes on them. The focus is on the helminths of seabirds, dominant parasites in coastal ecosystems. Their fauna in the Arctic is depauperate because of the lack of suitable intermediate hosts and unfavourable conditions for species with free-living larvae. An increasing proportion of crustaceans in the diet of Arctic seabirds would result in a higher infection intensity of cestodes and acanthocephalans, and may also promote the infection of seabirds with non-specific helminths. In this way, the latter may find favourable conditions for colonization of new hosts. Climate changes may alter the composition of the helminth fauna, their infection levels in hosts and ways of transmission in coastal communities. Immigration of boreal invertebrates and fish into Arctic seas may allow the circulation of helminths using them as intermediate hosts. Changing migratory routes of animals would alter the distribution of their parasites, facilitating, in particular, their trans-Arctic transfer. Prolongation of the seasonal 'transmission window' may increase the parasitic load on host populations. Changes in Arctic marine food webs would have an overriding influence on the helminths' circulation. This process may be influenced by the predicted decreased of salinity in Arctic seas, increased storm activity, coastal erosion, ocean acidification, decline of Arctic ice, etc. Greater parasitological research efforts are needed to assess the influence of factors related to Arctic climate change on the transmission of helminths.

Genomic diversity of cercarial clones of Himasthla elongata (Trematoda, Echinostomatidae) determined with AFLP technique.

The aim of this study was to reveal genomic diversity formed during parthenogenetic reproduction of rediae of the trematode Himasthla elongata in its molluskan host Littorina littorea. We applied amplification fragment length polymorphism (AFLP) to determine the genomic diversity of individual cercariae within the clone, that is, the infrapopulation of parthenogenetic progeny in a single molluskan host. The level of genomic diversity of particular cercariae isolates from a single clone, detected with EcoR1/Mse1 AFLP reaction, was significantly lower than the variability of cercariae from different clones. The presence of intraclonal genomic diversity indicates a nonsexual shuffle of alleles during parthenogenesis in the rediae of H. elongata. The obtained polymorphic AFLP fragments were long enough to detect the sequences that may be responsible for clonal genomic variability. Based on this, AFLP can be recommended as a tool for the study of genetic mechanisms of this variability.

[PATTERNS IN CIRCULATION AND TRANSMISSION OF MARINE BIRD PARASITES IN HIGH ARCTIC: A CASE OF ACANTHOCEPHALAN POLYMORPHUS PHIPPSI (PALAEACANTHOCEPHALA, POLYMORPHIDAE)].

This study, based on the materials on parasitic infection of marine birds and invertebrates in Frantz Josef Land (FJL) collected in 1991-1993, focussed on the acanthocephalan Polymorphus phippsi. We identified this parasite, confirmed its species status and analysed its circulation and transmission patterns in high Arctic. The causes of its erroneous identification as P. minutus in several studies were also examined. In contrast to P. minutus, the transmission of P. phippsi is realized in marine coastal ecosystems. Its' main intermediate host in the Arctic is the amphipod Gammarus (Lagunogammarus) setosus, commonin coastal. areas of the shelf zone throughout the Arctic basin. P. phippsi population in FJL and the entire European Arctic is on the whole maintained by a single obligate final host, the common eider Somateria mollissima. Prevalence (P) of P. phippsi in this bird reached 100 %, with the maximal infection intensity (IImax) of 1188 and the mean abundance (MA) of 492.1. Other species of birds found to be infected with P. phippsi (Arctic turn, black guillemot, purple sandpiper and several gulls) are facultative and/or eliminative hosts. The most heavily infected birds were Arctic terns (P = 72.7%, IImax = 227, MA = = 47.1), which contained single mature acanthocephalans. For one of the FJL regions, infections flows of P. phippsi through various host categories were calculated. Involvement of birds unrelated to the common eider into the circulation of P. phippsi is facilitated by their feeding character in the Arctic. While coastal crustaceans are abundant, fish food is relatively scarce (polar cod, snailfishes), and so amphipods make up a considerable part of the diet of marine birds in FJL, if not most of it, as for instance in case of Arctic tern. This promotes an easy entry of the larvae of crustaceans-parasitizing helminthes (cestodes and acanthocephalans, including cystacanths P. phippsi) into non-specific hosts and opens broad colonization possibilities. Besides acanthocephalans, the phenomenon of non-specific parasitism has been shown for some cestodes circulating in the Arctic coastal ecosystems. Similar conditions for helminths transmission might have formed in marine coastal refugia during the glacial periods of late Pliocene-Pleistocene. According to the Arctic refugium hypothesis of Hoberg and Adams, this promoted parasitic colonization of phylogenetically distant hosts using similar foraging resources. Thus, present-day transmission patterns of helminthes in high Arctic can be, in a way, considered as a model allowing us to witness various stages of helminthes' speciation by host-switching.

In vitro encystment of Himasthla elongata cercariae (Digenea, Echinostomatidae) in the haemolymph of blue mussels Mytilus edulis as a tool for assessing cercarial infectivity and molluscan susceptibility.

Infectivity of Himasthla elongata cercariae to mussels, their second intermediate hosts, and resistance by these hosts to infection were assessed on the basis of the cercariae's ability to encyst in mussel haemolymph in vitro. A series of experimental in vivo infections of mussels with batches of cercariae, each batch released from a different single infected mollusc and referred to as a clone (due to their shared genotype), demonstrated that the results of the in vitro tests corresponded to the actual indices of infectivity/susceptibility of the parasites and their hosts. Most cercarial clones had high infectivity, with a few clones having very high or, at the other extreme, very low infectivity. A similar pattern was revealed in mussel resistance to cercarial infection. Most of the molluscs tested were moderately susceptible to cercarial infection, but at each extreme a small fraction (less than 10%) displayed very high or very low susceptibility. It was shown that there were no totally compatible or totally incompatible 'cercaria clone/mussel' combinations. Results obtained are compared with the data on intra-population variability using the characters parasite infectivity/host compatibility for trematode/mollusc-first intermediate host associations. Results are made relevant to actual infection levels in mussel settlements at the White Sea.

[A case study of singular spectrum analysis application in parasitology: dynamics of prevalence of Cryptocotyle concavum and Bunocotyle progenetica trematode parthenitae in Hydrobia ventrosa snails at the White Sea].

In this study parasitological data were analyzed by different methods of revealing the structure of time series, namely auto-correlation analysis (ACA), Fourier spectrum analysis (SA) and singular spectrum analysis (SSA), and the results of these analysis were compared (SSA makes it possible to present non-stationary time series as a sum of independent components and to determine the contribution of each component into the dispersion of the initial series--Golyandina et al., 2001). This case study was based on the result of 10-year-long monitoring of changes in the prevalence of Cryptocotyle concavum and Bunocotyle progenetica trematode parthenitae in intertidal snails Hydrobia ventrosa at the White Sea (in total, 45 observations). ACA did not reveal any statistically significant oscillations in the analyzed series. The application of SSA and SA allowed us to reveal at least two quasi-periodical components. In addition, SSA made it possible to reveal a significant dome-shaped trend in the prevalence of B. progenetica parthenitae, which were described by SA as an oscillation with a period equal to the duration of the study, as well as to give proof that there was no trend in the changes of C. concavum parthenitae prevalence. The components (modes) extracted by the SSA described the changes in the prevalence better that the harmonics extracted by the SA. In particular, SSA modes (contrary to SA harmonics) reflected that the amplitude of oscillations of the B. progenetica prevalence increased as the prevalence grew. The sums of SSA modes correlated more with initial prevalence series that the sums of SA harmonics. A possible interpretation of the trends and modes extracted by the SSA in the light of the transmission features of the investigated trematode species in the study area was proposed.

[Clonal variability in longevity of the cercariae of Himasthla elongata (Trematoda: Echinostomatidae)].

The study was carried out on Himasthla elongata cercariae shed by infected Littorina littorea snails. The infected periwinkles were collected from the settlement with the low prevalence of H. elongata. As shown earlier with the use of AFLP (Amplified Fragment Length Polymorphisms) method, rediae groups in all the infected periwinkles of this settlement arise from the infection of a mollusc with a single miracidium. Therefore, the cercariae shed by an infected mollusc have the same genotype or, in other words, represent a clone. The LT50 (the time during which 50% of cercariae perish in the experimental dish) were measured experimentally for cercariae Himasthla elongata belong to different clones. The investigated parameter demonstrated a high level of interclonal variability. Two groups of cercarial clones were identified: one of them was characterized by the high level of intraclonal variability in LT50 and the second, by the low one. It is assumed that the observed heterogeneity may be stipulated by different degrees of mitotic recombinations during formation of different cercarial clones.

Morpho-functional specialization of the branching sporocyst of Prosorhynchoides borealis Bartoli, Gibson & Bray, 2006 (Digenea, Bucephalidae).

Sporocysts of Prosorhynchoides borealis were obtained from the marine bivalves Abra prismatica and studied using transmission electron microscopy. The sporocyst body consists of a mass of branching and intertwining hollow tubules that ramify through the host's digestive gland and gonads. This study investigated the ultrastructure of the sporocyst branches which comprise alternate distended areas (brood chambers) with a relatively thin body wall, narrower portions with a thicker body wall (constricted areas) and terminal regions. Pronounced differences between these areas were revealed in the structure of their tegument and body cavity lining, as well as in the cellular composition of the subtegumental layers. Body wall composition in distended areas was consistent with the specialization for cercarial nurture in the brood chambers. The structure of the constrictions suggested a dual role of nutrient absorption and physical separation of adjacent brood chambers. Two types of terminal region were identified, one specialized for the investigation and penetration of host tissues and the other, in which the germinal cells are formed, for cercarial production. The overall structure of the sporocyst branches helps explain why this linear modular system, i.e. brood chambers and constrictions continuously growing into the host tissue, enables the sporocyst's long-term existence and can continuously produce cercariae in numbers comparable with those produced by rediae and/or daughter sporocyst infrapopulations in other digeneans. The origin of the nuclei in the outer tegumental layer of some branching bucephalid sporocysts is also discussed.

[Clonal variability in expression of geo- and photoorientation in cercariae of Himasthla elongata (Trematoda: Echinostomatidae)].

The study was carried out on Himasthla elongata, a digenean common in the coastal ecosystems of the northern European seas. This species utilises intertidal prosobranchs Littorina spp. as the first intermediate host, bivalves (in the White Sea, Mytilus edulis) as the second intermediate host and gulls as the final host. The periwinkles Littorina littorea infected with H. elongata rediae (parthenogenetic generations) were sampled in the intertidal of the White Sea (66 degrees 20' N, 33 degrees 38' E) and used as the source of cercariae. Periwinkles were collected from the settlement with the low prevalence of H. elongata. As shown earlier with the use of AFLP (Amplified Fragment Length Polymorphisms) method, rediae groups in all the infected periwinkles of this settlement arise from the infection of a mollusc with a single miracidium. Therefore, the cercariae shed by an infected mollusc have the same genotype or, in other words, represent a clone. Photo- and geoorientation of cercariae originating from different clones and aged 1 h and 6 h were analysed separately. It was shown that in general the larvae of each clone followed the behavioural pattern characteristics of the species (positive geoorientation and negative photoorientation). However, the degree of expression of this typical behaviour was different in different clones. An especially high variability was observed in the manifestation of geoorientation (in several clones, most larvae demonstrated negative geoorientation). Differences in the distribution of cercariae in the illumination gradient were almost equally associated with the interclonal variability and the age of the larvae. On the whole, as the age of cercariae increased, the positive geoorientation became more prominent, whereas the ratio of cercariae with the typical (negative) photoorientation decreased. Statistical analysis revealed significant differences between the cercarial clones both in the initial manifestation of geo- and photoorientation and in the changes in the character of these reactions with the larval age. Taking into account that each cercarial clone investigated had the same genotype, it seems very likely that the interclonal differences noted in this study are hereditary. Maintenance of a rather high level of genetic polymorphism by the character "expression of orientation reaction" in trematode cercariae may enhance the chances for successful transmission of these larvae. Such variability increases the scale of cercarial dispersion in space and promotes the successful infection of the hosts, whose behaviour is also subject to intra- and inter-population variability. Besides, cercariae whose behaviour deviates from the basic behaviour of the species may play the role of the population's potential for colonisation of new species of animal hosts.

[Microphallus kurilensis sp. nov., a new species of microphallids from the pygmaeus species group (Trematoda, Microphallidae) from the coastal areas of Okhotsk and Bering Seas].

The pygmaeus-species group is composed of close related species from the genus Microphallus in which metacercariae develop inside daughter sporocysts without encystment. Infection of periwinkles Littorina (Neritremna) spp. with intramolluscan stages of a new species of this group (Microphallus kurilensis sp. nov.) was recorded on the coasts of Sakhalin and Kuril islands, north of the Sea of Okhotsk and Chukchi Peninsula (the Bering Sea). Application of molecular methods allowed us to establish that M. kurilensis metacercariae are conspecific with one of the morphotypes of microphallid adults obtained from the intestine of the Pacific common eider (Somateria mollissima v-nigrum), which was shot in the north of the Sea of Okhotsk (Galaktionov, Olson, and Blasco-Costa, in press). The adults of the same morphotype were recorded in the Pacific common eider from the northwestern part of the Bering Sea (Chukchi Peninsula). In the course of experimental infection of the slaty-backed gull Larus schistisagus chicks with metacercariae of M. kurilensis, few microphallid adults were obtained. These adults were identical in their morphology with specimens of the microphallid morphotype from the Pacific common eider, which had been identified as M. kurilensis based on molecular data. Morphological description of metacercaria and adult of M. kurilensis and list of their differences from the same developmental stages of other species from pygmaeus-group are provided. It is concluded that M. kurilensis is transmitted in the host system including periwinkle Littorina (Neritrema) and seaducks (predominately, Pacific common eider). Most probably, distribution of M. kurilensis is not limited by the north Asiatic coast but expanded to the North American coast of the Pacific Ocean.

[Effect of some pharmacological substances on the motility of the Cryptocotyle lingua cercaria (Heterophyidae)].

The effect of some biologically active substances (acetylcholine, serotonin, octopamine, sodium nitroprussid and FMRF-amide) on the motility of the Cryptocotyle lingua cercariae was studied. Solutions of FMRF-amide, octopamine, and sodium nitroprussid have no statistically significant influence on the motility of C. lingua. Acetylcholine and serotonin in solutions affected the motility through the prolongation of the active phase of swimming. Further research is required to elucidate the mechanisms underlying the cercarial motility.

CDC25 phosphatases as potential human oncogenes.

Cyclin-dependent kinases (CDKs) are activated by CDC25 phosphatases, which remove inhibitory phosphate from tyrosine and threonine residues. In human cells, CDC25 proteins are encoded by a multigene family, consisting of CDC25A, CDC25B, and CDC25C. In rodent cells, human CDC25A or CDC25B but not CDC25C phosphatases cooperate with either Ha-RASG12V or loss of RB1 in oncogenic focus formation. Such transformants were highly aneuploid, grew in soft agar, and formed high-grade tumors in nude mice. Overexpression of CDC25B was detected in 32 percent of human primary breast cancers tested. The CDC25 phosphatases may contribute to the development of human cancer.

[Description of the maritae and determination of the species status of Microphallus pseudopygmaeus sp. nov. (Trematoda: Microphallidae)].

Microphallus pseudopygmaeus belongs to the "pygmaeus" microphallids, a group of closely related species with homotypic two-host life cycle. This cycle involves one intermediate host (littoral or sublittoral gastropods, mostly of the genus Littorina), and bird-invading metacercariae develop within daughter sporocysts. In spite of the fact that the name Microphallus pseudopygmaeus is widespread in special literature, the original description of this species (Galaktionov, 1980) was performed in contravention of the International Code of Zoological Nomenclature. This article is aimed to correct the above defect. The maritae of Microphallus pseudopygmaeus were obtained from the common eider ducklings which were infected experimentally with metacercariae of Microphallus sp. I Galaktionov, 1980. These metacercariae were extracted from the naturally infected mollusks Littorina saxatilis collected at the Barents Sea coast. The valid description of the species M. pseudopygmaeus, its comparison with other representatives of the "pygmaeus" microphallids and synonymy are provided. Detailed analysis of digeneans from marine and coastal birds preserved in the collections of the Zoological Institute RAS (St. Petersburg, Russia) and the Natural History Museum (London, UK) showed that in some cases M. pseudopygmaeus was misidentified as M. pygmaeus. It has been shown that M. pseudopygmaeus has an amphiboreal-arctic distribution and the spectrum of its first intermediate hosts is unequally wide for digeneans. It includes 15 species of north-Atlantic and north-Pacific mollusks belonging to different families and even orders of Prosobranchia. At the same time, the range of final hosts of M. pseudopygmaeus is limited by benthophagous marine ducks, first of all the common eider.

E2F-Rb complexes assemble and inhibit cdc25A transcription in cervical carcinoma cells following repression of human papillomavirus oncogene expression.

Expression of the bovine papillomavirus E2 protein in cervical carcinoma cells represses expression of integrated human papillomavirus (HPV) E6/E7 oncogenes, followed by repression of the cdc25A gene and other cellular genes required for cell cycle progression, resulting in dramatic growth arrest. To explore the mechanism of repression of cell cycle genes in cervical carcinoma cells following E6/E7 repression, we analyzed regulation of the cdc25A promoter, which contains two consensus E2F binding sites and a consensus E2 binding site. The wild-type E2 protein inhibited expression of a luciferase gene linked to the cdc25A promoter in HT-3 cervical carcinoma cells. Mutation of the distal E2F binding site in the cdc25A promoter abolished E2-induced repression, whereas mutation of the proximal E2F site or the E2 site had no effect. None of these mutations affected the activity of the promoter in the absence of E2 expression. Expression of the E2 protein also led to posttranscriptional increase in the level of E2F4, p105(Rb), and p130 and induced the formation of nuclear E2F4-p130 and E2F4-p105(Rb) complexes. This resulted in marked rearrangement of the protein complexes that formed at the distal E2F site in the cdc25A promoter, including the replacement of free E2F complexes with E2F4-p105(Rb) complexes. These experiments indicated that repression of E2F-responsive promoters following HPV E6/E7 repression was mediated by activation of the Rb tumor suppressor pathway and the assembly of repressing E2F4-Rb DNA binding complexes. Importantly, these experiments revealed that HPV-induced alterations in E2F transcription complexes that occur during cervical carcinogenesis are reversed by repression of HPV E6/E7 expression.

Infection patterns in white sea blue mussels Mytilus edulis of different age and size with metacercariae of Himasthla elongata (Echinostomatidae) and Cercaria parvicaudata (Renicolidae).

Infection of mussels Mytilus edulis L. by 2 trematode species was studied in a natural intertidal population in the Chupa inlet of the White Sea. The prevalence of metacercariae of Himasthla elongata (Mehlis, 1831) and Cercaria parvicaudata (Stunkard & Shaw, 1931) in mussels reached 100% in 3 to 4 yr old molluscs and remained at this level in older individuals. Infection intensity increased evenly with the age of the molluscan host, showing a tendency to decrease only in the oldest (9 yr old) mussels. These patterns of age dynamics of prevalence and infection intensity were associated with accumulation of trematode larvae in the course of the molluscs' lives. Ability of metacercariae to exist in mussels for long periods (at least 2.5 yr) was verified in the course of an experiment, during which infected molluscs were kept in a subtidal net cage. Decrease of infection intensity in the oldest individuals may reflect selective mortality of the most severely infected molluscs. Among mussels of the same age, higher infection intensity values occurred in larger individuals. This may be due to an enhanced pumping rate in large molluscs, which increases the probability of cercariae, free-living trematode larvae, infecting them via water currents.

[A comparative analysis of the helminth fauna of kittiwake Rissa tridactyla (Linnaeus, 1758) and glaucous gull Larus hyperboreus Gunnerus, 1767 from different parts of the Barents Sea].

The article is based on the results of helminthological observations made on kittiwake Rissa tridactyla and glaucous gull Larus hyperboreus in 1991-2001 in different areas of the Barents Sea (Eastern Murman coast, Franz Josef Land, Novaya Zemlya, Spitzbergen). 18 helminth species (2 trematodes, 11 cestodes, 4 nematodes, and 2 acanthocephalans) were recorded in the kittiwakes and 19 (3 trematodes, 9 cestodes, 5 nematodes and 2 acanthocephalans) species were recorded in the glaucous gulls. Trematodes were absent in the birds collected at the Franz Josef Land and the northern island of Novaya Zemlya. 3 trematode species, namely Gymnophallus sp. (somateria?), Microphallus sp. 1 (M. pseudopygmaeus), and Cryptocotyle lingua were found in the glaucous gulls of western Spitzbergen. It was supposed that the life cycles of these parasites can be completed there. On the other hand, coastal ecosystems of Arctic archipelagoes turn out to be favourable for the transmission of some cestodes. This is closely connected with the regional traits in the marine bird diet, namely the increase of the amphipod (intermediate hosts of hymenolepidids and some dilepidids) and polar cod (supposed second intermediate host for some tetrabothriids) portion in Arctic. As a result, cestodes are the base of the helminth fauna of kittiwakes and glaucous gulls of the Barents Sea, by their species richness, prevalence and abundance. Nematodes and acanthocephalans were represented by a few species with low infection intensity. The main ecological factors affected the regional difference in the species richness and abundance of the helminths parasitising kittiwakes and glaucous gulls in the Barents Sea are proposed. Those are regional climatic features and regional traits in the behaviour and food priorities of birds, and also the distribution of the helminths intermediate hosts, invertebrates and fishes. The phenomenon of host specificity lowering with respect to the definitive host was recorded in some cestode species (Microsomacanthus diorchis, M. microsoma, and Arctotaenia tetrabothrioides) on the border of their distribution ranges, the coastal ecosystems of Arctic.

The glands of trematode cercariae of the family Microphallidae Travassos, 1920.

The ultrastructure of the glands of microphallid cercariae is described. The fully-formed cercariae possess only penetration glands opening onto the surface of the tegument. Cercarial embryos have tegumental glands transferring their secretion in the tegument. In all studied species two anterior pairs of penetration glands produce identical granules while two posterior pairs produce heterogeneous granules of various shapes and sizes. Tegument cells are divided into mucoid and two other types of tegument glands (TG1 and TG2). The mucoid glands of Microphallinae cercariae produce large fibrous granules, while those of Maritrematinae cercariae produce osmiophil ones. TG1 cells of Maritrema subdolum produce large rod-shaped granules and TG1 cells of Levinseniella brachysoma, Microphallus sp. and M. claviformis produce two types of osmiophil granules. TG2 produce small rod-shaped granules. The association of gland ultrastructure with their function and features of the microphallid life-history are discussed.

Screening for antimitotic compounds using the cdc25 tyrosine phosphatase, an activator of the mitosis-inducing p34cdc2/cyclin Bcdc13 protein kinase.

A universal intracellular factor, the "M phase-promoting factor" (MPF), triggers the G2/M transition of the cell cycle in all organisms. In late G2, it is present as an inactive complex of tyrosine-phosphorylated p34cdc2 and unphosphorylated cyclin Bcdc13. In M phase, its activation as an active MPF displaying histone H1 kinase activity originates from the specific tyrosine dephosphorylation of the p34cdc2 subunit by the tyrosine phosphatase p80cdc25. We describe here a colorimetric assay of recombinant human cdc25A tyrosine phosphatase used as a cell cycle-specific target to screen for antimitotic compounds. The glutathione-S-transferase/cdc25A tyrosine phosphatase fusion protein is produced in large amounts of Escherichia coli and easily purified by affinity chromatography on glutathione-agarose. Optimal purification, storage and assay conditions (concentrations of enzyme, p-nitrophenylphosphate and dithiothreitol; duration of assay) have been determined. Using this system we tested 15 compounds currently used in cancer treatment; none of them displayed any inhibitory activity. However, the assay detected the inhibitory activity of vanadate, a reported tyrosine phosphatase inhibitor. The simplicity, speed and possible extensive automation of this assay using an essential cell cycle-regulating component provide a highly specific mechanism-based screen for antimitotic drugs discovery.

Distribution patterns of marine bird digenean larvae in periwinkles along the southern coast of the Barents Sea.

An important component of the parasite fauna of seabirds in arctic regions are the flukes (Digena). Different species of digeneans have life cycles which may consist of 1 intermediate host and no free-living larval stages, 2 intermediate hosts and 1 free-living stage, or 2 intermediate hosts and 2 free-living larval stages. This study examined the distribution of such parasites in the intertidal zones of the southern coast of the Barents Sea (northwestern Russia and northern Norway) by investigating 2 species of periwinkles (Littorina saxatilis and L. obtusata) which are intermediate hosts of many species of digeneans. A total of 26,020 snails from 134 sampling stations were collected. The study area was divided into 5 regions, and the number of species, frequency of occurrence and prevalence of different digenean species and groups of species (depending on life cycle complexity) were compared among these regions, statistically controlling for environmental exposure. We found 14 species of digeneans, of which 13 have marine birds as final hosts. The number of species per sampling station increased westwards, and was higher on the Norwegian coast than on the Russian coast. The frequency of occurrence of digeneans with more than 1 intermediate host increased westwards, making up a larger proportion of the digeneans among infected snails. This was significant in L. saxatilis. The prevalence of different species showed the same pattern, and significantly more snails of both species were infected with digeneans with complicated life cycles in the western regions. In L. saxatilis, environmental exposure had a statistically significant effect on the distribution of the most common digenean species. This was less obvious in L. obtusata. The causes of changing species composition between regions are probably (1) the harsh climate in the eastern part of the study area reducing the probability of successful transmission of digeneans with complicated life cycles, and (2) the distribution of different final hosts.

[Intranuclear actin microfilaments in the oocytes of the common frog]. / Vnutriiadernye aktinovye mikrofilamenty v ootsitakh travianoi liagushki.

For identification and distribution of actin microfilaments in hand-isolated nuclei of R. temporaria oocytes (stage 6, according to Dumont, 1972) different methods were used: heavy meromyosin decoration, antiactin immunofluorescence with monoclonal antibodies, staining with rhodamine phalloidin, and electrophoresis in polyacrylamide gel. The nuclei of R. temporaria oocytes contain a considerable quantities of actin microfilaments which form intranuclear meshwork. Microfilaments are connected with the nucleoli, nucleolar RNP-complexes and nuclear envelope. Immunofluorescence with antiactin monoclonal antibodies reveals a strong staining of microfilaments and nucleoli. A slight staining of nucleoli is observed after the treatment of nuclei with rhodamine phalloidin. A specific role of intranuclear microfilaments in direct transport of nucleolar material from the nucleus into the oocyte cytoplasm, in stabilization of the karyosphere (the late diplotene oocyte complex of chromosomes with numerous nucleoli) is discussed in addition to its keeping in a definite region of the nucleus. A supposition is drawn on the functional significance of the connection between microfilaments and nuclear matrix. Based on our own and literature data, a conclusion is drawn, that the intranuclear filament actin may be one of the leading components in morpho-functional organization of the nucleus as the whole.