Diverse archaeal community of a bat guano pile in Domica Cave (Slovak Karst, Slovakia).

The molecular diversity of Archaea in a bat guano pile in Cave Domica (Slovakia), temperate cave ecosystem with significant bat colony (about 1600 individuals), was examined. The guano pile was created mainly by an activity of the Mediterranean horseshoe bat (Rhinolophus euryale) and provides a source of organic carbon and other nutrients in the oligotrophic subsurface ecosystem. The upper and the basal parts of guano surface were sampled where the latter one had higher pH and higher admixture of limestone bedrock and increased colonization of invertebrates. The relative proportion of Archaea determined using CARD-FISH in both parts was 3.5-3.9 % (the basal and upper part, respectively). The archaeal community was dominated by non-thermophilic Crenarchaeota (99 % of clones). Phylogenetic analysis of 115 16S rDNA sequences revealed the presence of Crenarchaeota previously isolated from temperate surface soils (group 1.1b, 62 clones), deep subsurface acid waters (group 1.1a, 52 clones) and Euryarchaeota (1 clone). Four of the analyzed sequences were found to have little similarity to those in public databases. The composition of both archaeal communities differed, with respect to higher diversity of Archaea in the upper part of the bat guano pile. High diversity archaeal population is present in the bat guano deposit and consists of both soil- and subsurface-born Crenarchaeota.

The phylogeny of diphyllobothriid tapeworms (Cestoda: Pseudophyllidea) based on ITS-2 rDNA sequences.

Phylogenetic analysis of sequences of the ITS-2 rRNA genes of 20 samples of pseudophyllidean cestodes of the family Diphyllobothriidae (Ligula, Digramma, Diphyllobothrium, and Schistocephalus) from different hosts and geographical regions revealed that: (1) the inclusion of ligulids (previously family Ligulidae) to the Diphyllobothriidae is correct; (2) Schistocephalus appears as the most basal taxon of the Diphyllobothriidae, well separated from Ligula and Digramma, thus supporting the validity of Schistocephalinae Dubinina, 1962; (3) Digramma belonged with samples of Ligula, thus suggesting its invalidity as a genus; and (4) isolates of Ligula, presumably belonging to Ligula intestinalis, are paraphyletic, indicating that this species may represent a complex of separate taxa. Our results indicate the necessity for a taxonomic revision of the family Diphyllobothriidae.

Effects of zopiclone on blood glucose level, serum lipid concentration and clot lysis time in normoglycemic and normolipidemic rats.

In normoglycemic and normolipidemic rats the i.p. injection of zopiclone induced an acceleration of fibrinolysis in a dose-dependent bell shaped manner and various changes of the blood glucose level. Total lipids, total cholesterol and triglyceride serum levels remained unaffected by doses of 1.25, 2.5 and 15.0 mg/kg, with the exception of the medium dose (5.0 mg/kg) and the next dose (10.0 mg/kg) which lowered them very significantly.

Acute effects of zopiclone on blood glucose level and serum lipids in hyperlipidemic rats. Interactions with PK 11195 and flumazenil.

UNLABELLED: Intraperitoneal administration of 5 mg/kg zopiclone a cyclopyrolone acting on the central benzodiazepine receptors was found to produce significant reduction of total lipids, total cholesterol and triglyceride in rats randered hyperlipidemic by intraperitoneal injection of Triton W-1339. Blood glucose level was also reduced. Flumazenil (10 mg/kg) potentiated the hypoglicemic effect of zopiclone but had no additional effect on serum lipids. PK 11195 (25 mg/kg) antagonized the hypolipidemic effects of zopiclone. IN CONCLUSION: 1. The central benzodiazepine receptors are not involved in the hypolipidemic activity of zopiclone. 2. The peripheral type benzodiazepine receptors are partly responsible, for the hypolipidemic activity of this cyclopirrolone. 3. The changes of blood glucose level induced by these drugs does not seem to be related to benzodiazepine receptors.

Acute effects of zopiclone on blood glucose level and serum lipids in hyperlipidemic rats.

In rats rendered hyperlipidemic by ip administration of Triton WR-1339, the ip administration of zopiclone at doses of 1.25, 2.5, 5.0, 7.5, 10.0 and 15.0 mg/kg, a cyclopyrrolone acting upon the central benzodiazepine receptors, induces very significant reductions of total lipids, total cholesterol, and triglycerides, at nearly all of the doses. The most active dose was 5.0 mg/kg. The blood glucose level was diminished by doses of 1.25, 2.5, 7.5 and 15.0 mg/kg and it was not changed by the rest of the other doses.

Cancer vaccines with emphasis on a viral oncolysate melanoma vaccine.

Biotherapy of malignant diseases has become the fourth treatment modality besides surgery, chemo- and radiotherapy. Whole cell melanoma vaccines with or without BCG and other adjuvants, purified ganglioside and shed antigens, recombinant viruses carrying tumor antigens, dendritic cells pulsed with antigenic peptides etc. are in clinical trials. Efficacious viral oncolysate vaccines induce the host to mount tumor-specific cytotoxic T-cell response and prevention of relapses is supported by clinical trials. The use of "polyvalent" whole cell vaccines vs. purified or genetically engineered single antigen vaccines is justified as i. only very few single tumor antigens are present in all tumors of a given histological type; and ii. antigen modulation occurs in tumors rendering them resistant to immune attack generated by vaccine against a single antigen. Thus polyvalent vaccines immunize against several antigens vs. against a selected antigen.

The origin and evolution of viruses (a review).

Viroids and prions might have existed early at the border of inanimate and living worlds. Most extant viruses can be characterized as derivatives of ancestors originating from episomal elements of prokaryotes (DNA phages) and later from eukaryotes. Retroviruses very likely originated from cellular retrotransposons. Retrograde evolution of some large viruses from obligatory intracellular bacteria is possible but the ontogenesis of extant bacteria does not include a viral form of existence (the filterable L forms are not viruses) and well-defined viruses do not regenerate back into vegetative bacterial forms. Biologists experimenting with the evolution of prokaryotic and eukaryotic ancient cells cannot ignore the earliest appearance of viruses within or outside the living matter. Viruses participated in and gave direction to the evolution and natural selection by coexisting with uni- and multicellular organisms for billions of years. The coevolution of viruses and their host cells is characterized by incessant attacks and counterattacks through gene rearrangements and mutations (induced in the virus by an immunological counterattack of the host or by transgression of species barriers by the virus) and recombinations. Recombinations occurred between viral and viral or viral and host genes. Acts of "molecular piracy" as practiced by ancient viruses endowed the virus with the expression of several host genes for the advantage of the virus in its replicative cycle and host-to-host spread. Probably the first immortalized and malignantly transformed cells were induced by viruses as viruses evolved anti-apoptotic measures. While infected cells resort to apoptotic death before the assembly of a new viral progeny, prominent are the anti-apoptotic measures viruses evolved in order to assure the completion of their full replicative cycle. Further, viruses may escape neutralization by host antibodies and may survive a counterattack by the host's T cells directed at virally infected cells of its own. Viruses may induce a form of tolerance and coexist with their host without inducing disease. Persistent and apparently or deceivingly apathogenic or even attenuated viral "quasi-species" populations may contain individual particles that regain virulence due to recombinations and/or gene rearrangements, especially when transgressing species barriers. Xenotropic viruses of animals may replicate in human cells and vice versa confounding experiments with xenotransplants or with use of veterinary viral vaccines for the treatment of human diseases.

Influence of the acute intraperitoneal administration of tetrazepam on blood glucose level and serum lipids in normoglycemic and normolipidemic rats.

The acute i.p. administration of tetrazepam (5, 7.5, 15 mg/kg) in normoglycemic and normolipidemic rats induced an increase in blood glucose level, a delay of fibrinolysis (when administered at the first two doses) and variable changes of serum lipids. These results are different from those obtained in hyperlipidemic rats treated with tetrazepam.

Effects of acute intraperitoneal administration of tetrazepam on blood glucose level and serum lipids in hyperlipidemic albino rats.

In rats rendered hyperlipidemic by the i.p. injection of Triton WR-1339, the i.p. administration of tetrazepam, a benzodiazepine (BZD) used mainly as a central myorelaxant, evoked significant reductions of serum lipids and blood glucose level. The dose-response curve was bell-shaped for serum lipids changes, whereas no clear dose-response relationship for blood glucose level modifications could be established. Tetrazepam was less active on serum lipids than other BZDs as diazepam or midazolam.

The effects of the intraperitoneal administration of midazolam on blood glucose level and serum lipids in streptozotocin-induced diabetes in rats.

In male Wistar rats rendered diabetic by streptozotocin administration, the intraperitoneal (i.p.) injection of midazolam (2.5-5.0 and 10 mg/kg) induced a significant reduction of hyperglycemia and hyperlipidemia. The plasma immunoreactive insulin level was slightly, but significantly increased. The lethality was diminished.

Effects of lormetazepam on glycemia and serum lipids in hyperlipidemic rats.

Lormetazepam (N-methyllorazepam) administered intraperitoneally to rats rendered hyperlipidemic by Triton WR-1339 induced an elevation of blood glucose level at all investigated doses. It brought about significant reduction of serum total lipids, total cholesterol and triglycerides. The dose-response relationship was bell-shaped. However, it presented two peaks, differing from the responses to other benzodiazepines (BZD) which were characterized by only one peak.

Effects of administration of temazepam on blood glucose and serum lipid levels in hyperlipidemic rats.

In rats rendered hyperlipidemic by the administration of Triton WR-1339 temazepam induced significant reductions of serum lipids. The effects was more reduced than of diazepam. The blood glucose level failed to be affected by most of the doses.

Effects of midazolam on glycemia and serum lipids in rats.

Midazolam administered ip. in albino rats (each group consisted from 10 animals rendered hyperdyslipidemic by the administration of Triton WR-1339) induced at most doses a significant reduction of glycemia (p < 0.001). However, the reduction of blood glucose level was outside of the dangerous level. Midazolam elicited also very significant decrease of the elevated serum lipids (p < 0.001). The pharmacological analysis of these phenomena by using the peripheral type benzodiazepine (BZD) receptors antagonist PK 11105, the central BZD receptor antagonist flumazenil and the purinergic P1 receptors antagonist aminophylline has shown that the effects on serum lipids were due, very probably to the stimulation of the peripheral type BZD receptors. Aminophylline seems to have the property to block the peripheral type BZD receptors. The effects on blood glucose level were very variable.

Monoclonal Antibodies to the [alpha]- and [beta]-Subunits of the Plant Mitochondrial F1-ATPase.

We have generated nine monoclonal antibodies against subunits of the maize (Zea mays L.) mitochondrial F1-ATPase. These monoclonal antibodies were generated by immunizing mice against maize mitochondrial fractions and randomly collecting useful hybridomas. To prove that these monoclonal antibodies were directed against ATPase subunits, we tested their cross-reactivity with purified F1-ATPase from pea cotyledon mitochondria. One of the antibodies ([alpha]-ATPaseD) cross-reacted with the pea F1-ATPase [alpha]-subunit and two ([beta]-ATPaseD and [beta]-ATPaseE) cross-reacted with the pea F1-ATPase [beta]-subunit. This established that, of the nine antibodies, four react with the maize [alpha]-ATPase subunit and the other five react with the maize [beta]-ATPase subunit. Most of the monoclonal antibodies cross-react with the F1-ATPase from a wide range of plant species. Each of the four monoclonal antibodies raised against the [alpha]-subunit recognizes a different epitope. Of the five [beta]-subunit antibodies, at least three different epitopes are recognized. Direct incubation of the monoclonal antibodies with the F1-ATPase failed to inhibit the ATPase activity. The monoclonal antibodies [alpha]-ATPaseD and [beta]-ATPaseD were bound to epoxide-glass QuantAffinity beads and incubated with a purified preparation of pea F1-ATPase. The ATPase activity was not inhibited when the antibodies bound the ATPase. The antibodies were used to help map the pea F1-ATPase subunits on a two-dimensional map of whole pea cotyledon mitochondrial protein. In addition, the antibodies have revealed antigenic similarities between various isoforms observed for the [alpha]- and [beta]-subunits of the purified F1-ATPase. The specificity of these monoclonal antibodies, along with their cross-species recognition and their ability to bind the F1-ATPase without inhibiting enzymic function, makes these antibodies useful and invaluable tools for the further purification and characterization of plant mitochondrial F1-ATPases.

Propheromones derived from codlemone.

Tricarbonyl [(8,9,10,11-η-8,10-dodecadien-l-ol] iron and the corresponding acetate prepared from 8,10-dodecadien-1-ol or its acetate, comprise the protected double-bond system of the molecule. After coming in contact with ambient oxygen, the iron complexes in question slowly release the corresponding pheromones of, for example, the codling moth,Cydia pomonella, and the pea moth,Cydia nigricana in highE,E purity and amounts that are sufficient for pest monitoring. A simple dispenser for propheromone application is proposed. Results of release rates in laboratory conditions and field trials are given.