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[Occurrence of Coxiella burnetti antibodies in cattle, sheep and small terrestrial mammals in the western region of Bohemia].

The prevalence of antibodies to C. burnetii was studied in the Karlovy Vary district (the Czech Republic) between 1987 and 1989. Sera of cows, sheep and heart eluates of small ground mammals were tested using the complement fixation test with C. burnetii antigen in phase II (Bodibion, Mevak Nitra, Slovakia). Titers greater or equal to 8 or 10 were considered as positive. Cows from two large-scale farms were tested four times (the average number of every sampling was 339 cows). Their seroprevalences varied from 0.3 to 4.3% and from 0.3 to 10.6% on the first and second farm, respectively. The highest seroprevalences were on both farms in July, 1987. No clinical signs of Q-fever were observed in cows from these farms. In May, 1988, sera of 96 sheep from two farms were tested. No antibodies to C. burnetii were detected. A total of 488 small ground mammals from 9 various localities was tested. Antibodies were ascertained in one Common Vole (Microtus arvalis) at a titer of 10 and in one Bank Vole (Clethrionomys glareolus) at a titer of 2560. At present, some strains of C. burnetii occur in cattle and small ground mammals in the study area but these strains do not cause the clinical Q fever as they did in the same area at the beginning of the fifties.

Animals↗

[Discovery of the pacemaker and heart conduction system in mammals. Fantasy and truth].

The first report about experimental investigations on the origin of the rhythms in the mammalian heart were published in 1888 by McWilliam. Similar experiments were described by Hering in several publications starting from 1900. The results led to the hypothesis of the myogenic genesis of the rhythms. However, in 1907, Hering stated that the origin of the stimulus in the mammal heart was probably a nervous function. In 1898, 1903, and 1906 Wenckebach described the function and the functional localization of the pacemaker in the heart of the intact man, and in 1906, he described the morphology of the pacemaker and its blood supply in the post mortem human heart. In the same year, Adam analyzed the localization of the "natural" pacemaker and the regions of the secondary pacemakers in the mammal heart. In 1907, Keith and Flack introduced the term "sino-auricular node". They quoted the findings of Wenckebach and Hering, and they mentioned McWilliam. A muscular connection between the right atrium and both ventricles of the mammal heart was found by Kent and by His in 1893. In 1907, Tawara called the structure of the origin of the a-v connection "atrio-ventricular node". It was, however, the scientific discovery of Tawara that the a-v pathway was connected with the muscles of the ventricles by the Purkinje fibers. He discovered that the pathway consists of muscles which are histologically divided and functionally different from the other heart muscles. It is a system of specialized muscles which conduct the stimulus for the contraction without contracting themselves.

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Genetic analysis of genomic methylation patterns in plants and mammals.

While it is now accepted that methylation of cytosine residues plays a role in various epigenetic phenomena in mammals and flowering plants, the involvement of methylation patterns in the regulation of normal development has remained a controversial and essentially untested issue in the 20 years since such a role was first proposed. Antisense suppression of a DNA methyltransferase in Arabidopsis and characterization of methylation-defective mutants of Arabidopsis have shown that perturbations of methylation patterns disrupt the development of plants, and targeted mutation of the murine gene that encodes the one known from of DNA methyltransferase has shown that methylation is required for cellular differentiation, genomic imprinting, and X chromosome inactivation in mammals. Ectopic expression of homeotic genes and homeotic transformations of floral organs in methylation-defective plants suggest that (in plants and perhaps mammals) heritable methylation patterns reinforce and may have supplanted heritable gene control mediated by chromosomal proteins of the Polycomb and trithorax groups. It is also possible that the developmental abnormalities are the result of ectopic gene expression caused by activation of transcription from nearby parasitic sequence elements that are normally repressed by methylation. Application of modern methods of genetic analysis promises to give definite answers to long-standing questions as to the roles and significance of genomic methylation patterns in normal development and genome defense.

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The occurrence of babesiasis affecting small terrestrial mammals and the importance of this zoonosis in Europe.

A short survey is given of the occurrence of babesiasis affecting small terrestrial mammals in some parts of Europe. Results obtained in studies of 7,038 small terrestrial mammals (42 species) from Czechoslovakia, Austria, Hungary, Yugoslavia and Bulgaria, show the distribution of Babesia microti in these countries. The authors found babesias in the following host species: Neomys anomalus, Clethrionomys glareolus. Microtus arvalis, M. agrestis, Apodemus agratius, A. flavicollis, A. sylvaticus and Mus musculus. The average incidence was very low-0.2% and varied in individual countries from negative to 0.7%. Problems of the natural foci of B. microti in Central Europe and discussed and hitherto recorded cases of Babesia infections in man are summarised. The epidemiological importance of the unspecific mammal hosts of B. microti is emphasised and the necessity of surveillance of this zoonosis in Europe is pointed out.

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Genetic linkage disequilibrium of deleterious mutations in threatened mammals.

The impact of negative selection against deleterious mutations in endangered species remains underexplored. Recent studies have measured mutation load by comparing the accumulation of deleterious mutations, however, this method is most effective when comparing within and between populations of phylogenetically closely related species. Here, we introduced new statistics, LDcor, and its standardized form nLDcor, which allows us to detect and compare global linkage disequilibrium of deleterious mutations across species using unphased genotypes. These statistics measure averaged pairwise standardized covariance and standardize mutation differences based on the standard deviation of alleles to reflect selection intensity. We then examined selection strength in the genomes of seven mammals. Tigers exhibited an over-dispersion of deleterious mutations, while gorillas, giant pandas, and golden snub-nosed monkeys displayed negative linkage disequilibrium. Furthermore, the distribution of deleterious mutations in threatened mammals did not reveal consistent trends. Our results indicate that these newly developed statistics could help us understand the genetic burden of threatened species.

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Ascorbic acid levels in the aqueous humor of nocturnal and diurnal mammals.

Ascorbic acid concentration is known to be very high in the aqueous humor of humans and most animals. The role it might play in ocular function is a subject of conjecture. Some have proposed that it might protect the eye against light-induced damage. We examined the aqueous humor from 22 species of mammals to determine the range of levels and to see if there was a correlation with behavior. A wide range of ascorbic acid levels in the aqueous humor was found. Most of the animals considered to be diurnal had higher ascorbic acid levels than the nocturnal animals. This would suggest that ascorbic acid in the aqueous humor may play a protective role in those animals who are most exposed to light. Regardless, any theory proposing a role for ascorbic acid in the eye in mammals must take the wide range of ascorbic acid levels into account.

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Location of the vector of jaw muscle force in mammals.

Previous work has suggested that the third molar lies just in front of the point where the resultant vector of jaw muscle force, estimated from dissections, intersects the tooth row. This point meets the jaw such that the vector is 30% of jaw length from the jaw joint. Thus, the vector divides the jaw in the ratio of 3:7 when measurements are taken perpendicular to the vector. In practice, however, distances along mammalian jaws are typically measured on an easily determined line such as a line from one end of the tooth row to the other. The position of the jaw joint is then projected onto this line. As a rule, such a line is not perpendicular to the vector and so the distance from the projection of the joint, out to the rear of the third molar (and the vector's intersection), is different in different mammals. Rarely is this distance 30% of total jaw length. However, when the location of the vector's intersection is measured along the tooth row, this position varies directly with the inclination of the vector; a vector inclined posteriorly intersects the tooth row far from the projection of the joint and an anterior vector's intersection is relatively close. Only a vector perpendicular to the line from one end of the tooth row to the other intersects at 30%. This obvious point suggests a way to test the above hypotheses when the inclination of the vector is not known exactly. The predicted relationship between the distance to the molar, as a percentage of the total jaw length, and the approximate inclination of the vector derived from muscle weights (posterior or anterior depending on whether the temporalis or the masseter/pterygoid, respectively, is dominant) was observed in a sample of 46 different mammals.

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New approach to cell lineage analysis in mammals using the Cre-loxP system.

The Cre-loxP site-specific recombination system was used for cell lineage analysis in mammals. We constructed an expression plasmid, pCETZ-17, which consists of cytomegalovirus enhancer/chicken beta-actin promoter (CAG), a portion of the rabbit beta-globin gene, loxP-flanked DNA sequence (containing enhanced green fluorescent protein (EGFP) cDNA), and lacZ gene encoding E. coli beta-galactosidase (beta-gal). When circular pCETZ-17 plasmid DNA was microinjected into the pronuclei of fertilized eggs and these eggs were allowed to develop to two-cell stage, 62.8% (59/94) of the two-cell embryos exhibited distinct fluorescence in one or both blastomeres, but never expressed lacZ protein, as evaluated by histochemical staining with X-Gal, a substrate for beta-gal. When both circular plasmids, pCETZ-17 and pCAG/NCre (containing CAG and DNA sequences encoding nuclear location signal and Cre), were co-injected into fertilized eggs, almost all (87.0%, 47/54) embryos exhibited low or no fluorescence, but 51.9% (27/52) exhibited positive staining for beta-gal activity. This indicates that transient expression of the Cre recombinase gene removed the loxP-flanked DNA sequence in pCETZ-17 and then caused expression of the downstream lacZ sequence. We next microinjected pCETZ-17 into the pronuclei of fertilized eggs, cultured these injected eggs for 1 day, and collected only two-cell embryos expressing EGFP in both blastomeres. One blastomere of the EGFP-expressing two-cell embryos was microinjected with pCAG/NCre, and these treated embryos were cultured for 1 day up to four-cell stage. When the developing four-cell embryos were subjected to staining with X-Gal, cell lineage-related staining pattern for beta-gal activity was observed in most (77.8%, 7/9) embryos. These findings were further confirmed using two-cell embryos derived from a transgenic mouse line carrying CETZ-17 transgene. Thus, our system, which is based on transient expression of the Cre recombinase gene directly introduced into nuclei of embryonic cells by microinjection, is a powerful means for cell lineage analysis in mammals.

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Conserved developmental algorithms during thalamocortical circuit formation in mammals and reptiles.

The general patterns of early thalamocortical development follow a similar sequence in all mammals. Thalamocortical projections descend through the ventral thalamus, advance in the internal capsule amongst cells which already possess dorsal thalamic projections, then reach the cerebral cortex by associating with subplate cells and their early corticofugal projections. Initially, the thalamic projections pause in the internal capsule and subplate layer. The interactions of the thalamocortical projections with the early generated, largely transient cells of the subplate, marginal zone, internal capsule and ventral thalamus are believed to play a crucial role in the organized deployment of thalamic projections and establishing a functional cortical architecture. Selective fasciculation, contact guidance and release of neurotrophic factors are thought to play roles in the development of thalamocortical projections. These ideas are obtaining support from recent work on reeler and other strains of mice. The evolutionary origin of these largely transient cells and the overlying logic of early developmental steps are not understood. The behaviour of the thalamocortical and corticothalamic projections at the corticostriatal junction is particularly puzzling. The comparison of early forebrain development in mammals and reptiles is beginning to reveal highly conserved cellular and molecular interactions during early thalamocortical development and to reveal homologies between telencephalic subdivisions.

Algorithms↗

Photic influences on the developing mammal.

In adult mammals, the daily light-dark cycle acts via the retinohypothalamic pathway to entrain the circadian clock in the suprachiasmatic nuclei (SCN) and to communicate information about daylength to photoperiodic species. Studies in rats show that during late fetal and early neonatal life, before the retinohypothalamic pathway has innervated the SCN, the maternal circadian system entrains the timing of the developing clock to prevailing lighting conditions. Although the nature of the maternal output signal(s) used to entrain the developing clock has not been elucidated, the maternal SCN are a necessary component of maternal entrainment during both prenatal and postnatal life. Maternal entrainment of the fetal and neonatal clock thus ensures that the developing circadian system is synchronized to the outside world until maturation of the retinohypothalamic pathway permits direct photic entrainment. The maternal circadian system is not only necessary for entrainment of the developing circadian system, but recent studies suggest it may also provide the immature mammal with important photoperiodic information. In the montane vole (Microtus montanus) and the Djungarian hamster (Phodopus sungorus), the prenatal photoperiod affects postnatal photoperiodic responses, and cross-fostering experiments show that this information about daylength is perceived by the fetus. This prenatal information, in conjunction with postnatal perception of photoperiod, allows the developing animal to determine which way the season is changing and to modify the rate of reproductive maturation accordingly.

Aging↗

Quantitative study on the relation between structural and functional properties of the hearts from three different mammals.

The ultrastructural quantitative composition of left ventricular cardiac myocytes from isolated Langendorff-perfused hearts was studied in three different mammals (rabbit, guinea pig, and rat). Volume densities of mitochondria, myofibrils, and unspecified cytoplasm were determined using morphometry and were compared to functional parameters including left ventricular developed pressure (LVDP), contractility (dP/dt), heart rate, TTI (tension-time index, an index of oxygen consumption), and relative heart mass (H/B) obtained from these hearts. Each of the mammals was found to possess a very specific and characteristic quantitative composition of cardiac myocyte. Cardiac myocytes contained 26.8% mitochondria and 56.3% myofibrils in rabbits, 25.8% mitochondria and 60.9% myofibrils in guinea pigs, and 27.7% mitochondria and 58.1% myofibrils in rats. The LVDP, contractility, heart rate, and TTI were quite different among species. However, there were close correlations between the mitochondrial volume density and the LVDP (p < 0.05), and between the mitochondrial volume density and the TTI (p < 0.05), in any group of the animals. It is concluded that the mitochondrial volume density is a good indirect indicator of function of cardiac muscle related to oxidative capacity.

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Trophoblast and hypoblast in the monotreme, marsupial and eutherian mammal: evolution and origins.

The pregastrula stage mammalian conceptus consists of both embryonic and non-embryonic components. The latter forms the bulk of the tissues, provides nutrition for the developing embryo and also contributes developmental signals that influence events within the embryo itself. Understanding the origins and relationships between the embryonic and extraembryonic cell lineages is thus central to understanding development in mammals. Despite the apparent gross differences in early developmental strategy and form, the conceptuses of eutherian, marsupial and monotreme mammals show some remarkable similarities in the lineage allocation to trophoblast and hypoblast and in the emergent properties of the two cell types. We suggest that the gross differences can be explained by two relatively small evolutionary timing changes affecting cell adhesion patterns and the polarisation of developmentally significant information. These changes result in the conversion of a unilaminar blastocyst to a morula form composed of blastomeres with increased regulatory capacity.

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Visual factors in sound localization in mammals.

The ability of mammals to localize sound varies widely among species. During the past decade, evidence has accumulated that this variation cannot be accounted for simply on the basis of the availability of the physical cues for locus. Evidence is presented that a major factor in sound localization is the need to direct the field of best vision to a sound source for further scrutiny. Thus, species with broad fields of best vision (such as visual streaks) require less accurate information regarding the location of a sound source than do species with very narrow fields of best vision (such as foveae). To support this suggestion, data are reported for the width of the field of best vision in the form of retinal ganglion cell isodensity contours for thirteen species of mammals. The possible contribution of other factors including binocular fields, visual acuity, and the degree to which a species is predatory in lifestyle, is also examined.

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Connections of somatosensory cortex in megachiropteran bats: the evolution of cortical fields in mammals.

The cortical connections of the primary somatosensory area (SI or 3b), a caudal somatosensory field (area 1/2), the second somatosensory area (SII), the parietal ventral area (PV), the ventral somatosensory area (VS), and the lateral parietal area (LP) were investigated in grey headed flying foxes by injecting anatomical tracers into electrophysiologically identified locations in these fields. The receptive fields for clusters of neurons were mapped with sufficient density for injection sites to be related to the boundaries of fields, and to representations of specific body parts within the fields. In all cases, cortex was flattened and sectioned parallel to the cortical surface. Sections were stained for myelin and architectonic features of cortex were related to physiological mapping and connection patterns. We found patterns of topographic and nontopographic connections between 3b and adjacent anterior parietal fields 3a and 1/2, and fields caudolateral to 3b (SII and PV). Area 1/2 had both topographic and nontopographic connections with 3b, PP, and SII. Connections of SII and PV with areas 3b, 3a, and 1/2 were roughly topographic, although there was clear evidence for nontopographic connections between these fields. SII was most densely connected with area 1/2, while PV was most densely connected with 3b. SII had additional connections with fields in lateral parietal cortex and with subdivisions of motor cortex. Other connections of PV were with subdivisions of motor cortex and pyriform cortex. Laminar differences in connection patterns of SII and PV with surrounding cortex were also observed. Injections in the ventral somatosensory area revealed connections with SII, PV, area 1/2, auditory cortex, entorhinal cortex, and pyriform cortex. Finally, the lateral parietal field had very dense connections with posterior parietal cortex, caudal temporal cortex, and with subdivisions of motor cortex. Our results indicate that the 3b region is not homogeneous, but is composed of myelin dense and light regions, associated with 3b proper and invaginations of area 1/2, respectively. Connections of myelin dense 3b were different from invaginating portions of myelin light area 1/2. Our findings that 3b is densely interconnected with PV and moderately to lightly interconnected with SII supports the notion that SII and PV have been confused across mammals and across studies. Our connectional evidence provides further support for our hypothesis that area 1/2 is partially incorporated in 3b and has led to theories of the evolution of cortical fields in mammals.

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Muscle regeneration in amphibians and mammals: passing the torch.

Skeletal muscle in both amphibians and mammals possesses a high regenerative capacity. In amphibians, a muscle can regenerate in two distinct ways: as a tissue component of an entire regenerating limb (epimorphic regeneration) or as an isolated entity (tissue regeneration). In the absence of epimorphic regenerative ability, mammals can regenerate muscles only by the tissue mode. This review focuses principally on the regeneration of entire muscles and covers what is known and what remains to be elucidated about fundamental mechanisms underlying muscle regeneration at this level.

Aging↗

Mouse histocompatibility-related genes are not conserved in other mammals.

In addition to the genes for classical H-2 antigens, the H-2 complex of the mouse contains numerous homologous genes belonging to several distinct families. It is not known whether they have any functions. To address this question, I have investigated whether these genes are separately conserved in evolution. Subcloned 5' gene segments, encoding the variable domains, were used as hybridisation probes on genomic DNA blots of various mammals. Only the largest gene family, which includes the classical H-2/HLA genes, is detectable in humans and other mammals. The other gene families, including Qa-2 and T1a, are not conserved even in rodents. Most or all of their coding sequences are therefore redundant.

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Developmental aspects of X chromosome inactivation in eutherian and metatherian mammals.

The single active X principle has served for two decades as a focal point for research on the cyclic activation and inactivation of gene loci. Differences in X chromosome inactivation patterns of eutherian and marsupial mammals provide probes for investigating the mechanisms of the X inactivation process. In eutherian mammals, the X chromosome is inactivated early in meiotic prophase in males and remains inactive throughout the rest of spermatogenesis. During meiosis in females, the inactive X chromosome is activated so that both X chromosomes are active in oocytes. During the early cleavage divisions of female embryos, the paternally derived X is activated. It and the maternally derived X remain active until differentiation begins in early embryogenesis. At that time, the paternally derived X is inactivated in cells that give rise to extraembryonic membranes, whereas a random process determines which X chromosome is inactivated in cells that give rise to the embryo itself. Although less is known about developmental aspects of X inactivation in female marsupials, it is clear that the paternal X is preferentially inactive in postembryonic somatic cells. Furthermore, the paternal X is partially active at some loci in some cell types, indicating that it is not regulated as a single unit. The successful adaptation of a small (80-150 g), fecund marsupial to simple laboratory conditions now enables extensive experimentation on the large number of marsupials at various developmental stages. This capability, coupled with the application of newly developed cellular and molecular techniques to questions about X chromosome inactivation, shows great promise for advancing our understanding of the mechanisms that control the cyclic behavior of X chromosome activity.

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Production of genetically identical sets of mammals: cloning?

In the past few years, there has been phenomenal progress in producing genetically identical mammalian multiplets by asexual means. The simplest method is to divide embryos into two, three, or four parts, and transfer the parts to surrogate mothers for gestation to term. Another approach is to inject nuclei from totipotent embryonic cells into fertilized one-cell embryos and to remove both male and female pronuclei. When these embryos reach the blastocyst stage, nuclei from their totipotent cells can be removed and injected into one-cell embryos, thus amplifying the process ad infinitum. Subsets of these embryos can be stored in liquid nitrogen to make genetic material for additional copies available indefinitely. Some of these techniques are fairly efficacious, others are not. No reliable techniques are available for making genetic copies of nonhomozygous adult mammals (or other vertebrates) unless samples of embryonic cells were cryopreserved when the individuals were embryos. However, injection of nuclei of spermatogonia into enucleated one-cell embryos is a promising strategy. The genetic totipotency of these nuclei is obvious since they become gamete nuclei. Moreover, the birth of mice from the diploidized genetic material of a single X-bearing spermatozoan after removal of the female pronucleus from a one-cell fertilized embryo suggests that we should eventually be able to make genetic copies of adult male mammals. This approach would not work for adult females because their oogonia have all become oocytes, and the DNA configuration in oocytes is not identical to somatic cells due to meiotic crossing over.

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