Some diagnostic, biologic and morphologic characteristics of Francisella tularensis strains isolated from the ticks Ixodes ricinus (L.) in the Prague agglomeration

Four infectious agents were isolated from the ticks Ixodes ricinus (L.) collected in the recreational area and park-forest of Prague. On the basis of cultivation, staining, biochemical, serologic properties, pathogenicity for animals and histological tests they were identified as Francisella tularensis with the following features: they are short, gram-negative rods of approximate dimensions of 0.3 X 0.8 micron, growing in enriched media after 3-4 day incubation at 37 degrees C. They form small circular, at first transparent, later greyish turbid colonies with regular rims. They are little active biochemically. They are susceptible to streptomycin and some broad spectrum antibiotics. They react positively with tularemic serum, but in lower titres than those in which this serum reacts with standard antigen. The microbes are highly pathogenic for mice, guinea-pigs, young rats, in which a massive bacteriemia occurs before death, but they do not kill rabbits. They multiply well in chick embryo, but do not grow in cell or tissue cultures. The most important histologic changes were observed in liver and spleen of mice. No pathologic changes were found in brain, lungs, heart, kidneys. Necroses were found in liver and in their marginal zones the microbes were present. Conspicuous were changes in numerous hepatocytes which became enlarged due to microbial multiplication and finally transformed into "sacs" packed with microbes. Histological and electronoptical examination showed that these are intracellular parasites fringed with a light lytic zone. Discussed is the problem to what extent the properties of the isolated strains are typical of F. tularensis as well as the importance of their detection from the aspect of epidemiology and differential diagnostics.     

A fossil noctuid moth egg from the late cretaceous of eastern north america

A moth egg assignable to the family Noctuidae (Lepidoptera) is described from 75-million-year-old sediments from Martha's Vineyard, Massachusetts. This sample, which extends the fossil record of this farnily and modern heteroneuran moths back to the Cretaceous, may provide insight into the coevolution of moths and flowering plants, as well as have implications for the evolution of bats.     

Facile catalyst separation without water: fluorous biphase hydroformylation of olefins

A novel concept for performing stoichiometric and catalytic chemical transformations has been developed that is based on the limited miscibility of partially or fully fluorinated compounds with nonfluorinated compounds. A fluorous biphase system (FBS) consists of a fluorous phase containing a dissolved reagent or catalyst and another phase, which could be any common organic or nonorganic solvent with limited or no solubility in the fluorous phase. The fluorous phase is defined as the fluorocarbon (mostly perfluorinated alkanes, ethers, and tertiary amines)-rich phase of a biphase system. An FBS compatible reagent or catalyst contains enough fluorous moieties that it will be soluble only or preferentially in the fluorous phase. The most effective fluorous moieties are linear or branched perfluoroalkyl chains with high carbon number; they may also contain heteroatoms. The chemical transformation may occur either in the fluorous phase or at the interface of the two phases. The application of FBS has been demonstrated for the extraction of rhodium from toluene and for the hydroformylation of olefins. The ability to separate a catalyst or a reagent from the products completely at mild conditions could lead to industrial application of homogeneous catalysts or reagents and to the development of more environmentally benign processes.     

Histochemistry of gland cells of Eurytrema pancreaticum cercariae.

The cercariae of Eurytrema pancreaticum (Janson, 1889) possess four types of gland cells - proper cystogenic, penetration, ventral and dorsal gland cells. The secretion of ventral and dorsal gland cells is released into the tegument. The proper cystogenic gland cells are the largest and their contents serve for the formation of the cyst wall of metacercariae in the second intermediate host. The secretion of proper cystogenic gland cells contains besides neutral mucosubstances also acid mucosubstances with both carboxyl- and sulphogroups digestible with beta-glucuronidase. The secretion of penetration gland cells contains neutral mucosubstances and proteins with tyrosine, tryptophan and SS groups. The ventral gland cells contain mostly acid mucosubstances with sulphogroups, which are digested with beta-glucuronidase, and proteins with tyrosine, tryptophan and SH groups. The rudimentary dorsal gland cells contain a small amount of acid mucosubstances. The whole tegument of cercariae and the two main collecting canals of the excretory system exhibit a high alkaline phosphatase activity. The nerve ring and the main nerve truncs contain proteins with SH groups and hydrophilic lipids and exhibit a cholinesterase activity. The suckers contain a larger amount of glycogen.     

Tissue reaction to Cysticercus bovis in the lung of artificially infected cattle

The tissue reaction to Cysticercus bovis in the lung of cattle with an experimental infection was an inflammatory rim originating in the immediate vicinity of the cysts. The cysts recovered at days 83 and 102 p.i. contained living cysticerci. The rim was composed either of a layer of high histiocytes organized in palisades (at day 83 p.i.), or a lyer of flat histiocytes (at day 102 p.i.). The outer layer of the rim consisted of fibroblasts, reticular cells and a different number of eosinophil- and neutrophil luekocytes. On the periphery, the rim was formed by granulation tissue infiltrated with lymphoplasmocytes. At the border between the layers of the inflammatory rim there were conspicuous foci of a necrotic appearance typical of a tissue reaction to C. bovis.     

The effect of aging of milk samples on the accuracy of infrared analysis of milk composition]

The effect of a decrease (and/or fermentation) in the lactose content during milk storage under different conditions was investigated on the accuracy of the results obtained on a Milko-Scan apparatus to contribute to the present knowledge of this problem. The results were in agreement with some results cited in the literature. These wavelengths are used for infrared spectrophotometry on the above apparatus: for fat 3.48 microns, for proteins 6.46 microns and for lactose 9.60 microns. Bulk milk samples used for the tests were untreated or treated with potassium dichromate, bronopol, sodium azide and Milkofix at the temperatures of storage in darkness 20 degrees C and 4 degrees C. The differences against the reference values (measured on the first day) were determined and evaluated in milk composition and characteristics as arising during milk storage. These differences were used in form of either cumulative means of differences (Figs. 1 to 5) or individual differences (Fig. 8). In the first part significant correlation coefficients (P less than 0.001) were calculated for the relationship between the variations of lactose content and the fat and protein contents: r = -0.59 and/or -0.73 (Figs. 6 and 7). This suggests that the decrease in the lactose content by 0.10% recorded by the infrared analysis and caused by lactose decomposition is accompanied by a "seeming" increase in the fat and protein content by about 0.04%. In the second part the correlation coefficients for the fat and protein contents r = -0.96 and -0.96 (P less than 0.001; Figs. 9 and 10; Tab. II) were calculated on the basis of an observation of the lactose decrease in an untreated milk sample (20 degrees C for 28 hours). These coefficients are somewhat different from the preceding ones; this is due to the lower homogeneity of the first set where the milk samples were treated in a different way, but the coefficients confirm the same conclusions. The values of the correlation coefficients for the dependence between the development of the acquired titratable acidity (SH) and the variations of fat (F), protein (P) and lactose (L) contents were as follows: r = 0.95; 0.95; -0.99 (P less than 0.001; Figs. 12, 13; Tab. II). Thus the above-mentioned "seeming" increase in the F and P contents can be explained to the extent of 92.2% from the decrease in the L content, which also causes the increase in titratable acidity to the extent of 98.0%.(ABSTRACT TRUNCATED AT 400 WORDS)     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

Effect of fertilizers on Taenia saginata Goeze, 1782 egg viability in vitro

The effect of seven fertilizers on the viability of Taenia saginata eggs was tested in vitro. The most destructive effect was produced by lime-nitrogen, in which the eggs survived only for 24 hours in the substance and for two days in a saturated solution. In the limestone the eggs survived for three days.