Host B chromosomes as potential sex ratio distorters of intestinal nematode infrapopulations in the yellow-necked mouse (Apodemus flavicollis).

The yellow-necked mouse, Apodemus flavicollis, can be considered as a model for genetic polymorphism produced by the frequent presence of supernumerary or B chromosomes (Bs). Host genetic background is rarely taken into account in studies of parasite sex ratio. The main aim of this study was to investigate the range of infrapopulation sex ratios for nematode parasites of the yellow-necked mouse and to determine which factors most influence variation in parasite sex ratios. Six nematode species found in the collected yellow-necked mice were analysed. We confirmed the predominant pattern of female-biased sex ratios in vertebrate parasite infrapopulations. The presence of B chromosomes in host genomes played an important role in infrapopulations of Heligmosomoides polygyrus, Syphacia stroma and Trichuris muris, as hosts with B chromosomes carried a higher proportion of males. The relative increase of males in infrapopulations could result from a shift in parasite life history strategy, induced by adaptation to the specific host genotypes (Bs present). In a meta-analysis with previously published data, the sex determination system was demonstrated to play a significant role in nematode sex ratio variation, as well as specific life history patterns, such as the place of egg hatching.

Is there a Host Sex Bias in Intestinal Nematode Parasitism of the Yellow-necked Mouse (Apodemus Flavicollis) at Obedska Bara Pond, Serbia?

Fifty-one yellow-necked mice from the Obedska bara locality were analysed for the presence of intestinal nematode parasites in order to assert whether there was a host sex bias in infection. Previous research indicated that males would be the more infected sex, either due to the immunosuppressive effect of testosterone or their different allocation of resources towards immune defence. Quantitative infection parameters were compared between host sexes for all nematode species and nematodes in general. In addition, the influence of host sex, age, total body length, body mass and presence of other nematode species on parasite abundance was analysed. No statistically significant differences between males and females were noted for any of the studied quantitative parameters, leading to an absence of sex-biased parasitism in this study.

Diversity of nematodes in the yellow-necked field mouse Apodemus flavicollis from the Peripannonic region of Serbia.

Up to six nematode species were identified from 86 specimens of the yellow-necked field mouse Apodemus flavicollis from three mountainous localities known as Avala, Cer and Liskovac in Serbia. The highest prevalence of infection of 97% was recorded from Mt. Avala. Only one nematode species, Syphacia frederici, occurred in all three localities. There was complete overlap in nematode species from Mts. Avala and Liskovac, whereas the taxonomic distinctness of Mt. Cer was seen in the presence of the insect-transmitted species Rictularia proni. Locality was a statistically significant factor in all the best-fitted generalized linear models of variation in abundances. The highest level of both species richness and parasite alpha diversity (Shannon's H= 1.47) was found in the easternmost Mt. Liskovac, whereas the diversity indices were lowest for the westernmost Mt. Cer (Shannon's H= 0.48). In view of this geographical difference, the beta diversity indices were calculated along a west to east longitudinal gradient.

Two-module organization of the mandible in the yellow-necked mouse: a comparison between two different morphometric approaches.

Mandibles of yellow-necked mouse (Apodemus flavicollis) were used to explore modularity. We tested a biological hypothesis that two separate modules (alveolar region and ascending ramus) can be recognized within the mandible. As a second research goal, we compared two different morphometric procedures under the assumption that methodological approaches that use either geometric or traditional morphometric techniques should give similar results. Besides confirmation of the predicted hypothesis of modularity, the application of both approaches revealed that: (i) modularity was somewhat more evident when it was analysed on the asymmetric (fluctuating asymmetry, FA) than on the symmetric (individual variation) component of variation; (ii) there is correspondence in the patterns of individual variation and FA, which indicates that integration of mandibular traits among individuals is primarily due to direct developmental interactions; and (iii) allometry does not obscure the hypothesized modularity for individual variation or for FA. In addition, traditional morphometric method allowed us to check whether allometry influenced each module to the same extent and to conclude that the ascending ramus is more heavily influenced by general size than the alveolar region. In studies of modularity, differences in methods can lead to discrepancies in the results, and therefore, some caution is required when comparing findings from different investigations. The substantial agreement between our results provides evidence that, when considering two-module organization of the mouse mandible, direct comparison among studies that use the methods applied herein is, in great part, reliable.

The origin of B chromosomes in yellow-necked mice (Apodemus flavicollis)-Break rules but keep playing the game.

B chromosomes (Bs) are known for more than hundred years but their origin, structure and pattern of evolution are not well understood. In the past few years new methodological approaches, involving isolation of Bs followed by whole DNA amplification, DNA probe generation, and fluorescent in situ hybridization (FISH) or the B chromosome DNA sequencing, has allowed detailed analysis of their origin and molecular structure in different species. In this study we explored the origin of Bs in the yellow-necked wood mouse, Apodemus flavicollis, using generation of microdissected DNA probes followed by FISH on metaphase chromosomes. Bs of A. flavicollis were successfully isolated and DNA was used as the template for B-specific probes for the first time. We revealed homology of DNA derived from the analyzed B chromosomes to the pericentromeric region (PR) of sex chromosomes and subtelomeric region of two pairs of small autosomes, but lower homology to the rest of the Y chromosome. Moreover, all analysed Bs had the same structure regardless of their number per individual or the great geographic distance between examined populations from the Balkan Peninsula (Serbia) and Eastern Europe (south region of Russia and central Belarus). Therefore, it was suggested that B chromosomes in A. flavicollis have a unique common origin from the PR of sex chromosomes, and/or similar evolutionary pattern.

Genetic analysis of Dobrava-Belgrade virus from western Serbia--a newly detected focus in the Balkan Peninsula.

Dobrava-Belgrade virus (DOBV) is a hantavirus species that causes the most severe form of haemorrhagic fever with renal syndrome (HFRS) in Europe. DOBV has been detected in three Apodemus rodents: A. flavicollis, A. agrarius and A. ponticus. These emerging viruses appear throughout the Balkan Peninsula including Serbia as its central part. In this study, we examined the seroprevalence, molecular epidemiology and phylogenetics of DOBV from A. flavicollis captured at six Serbian localities. Furthermore, we applied microsatellite typing of host animal genome to analyse the role of host kinship in DOBV animal transmission. The overall IgG seropositivity rate over 3 years (2008-2010) was 11.9% (22/185). All seropositive samples were subjected to RT-PCR and DNA sequencing for S and L genome segments (pos. 291-1079 nt and 2999-3316 nt, respectively). DOBV was genetically detected in three samples from mountain Tara in western Serbia, a newly detected DOBV focus in the Balkans. No sequence data from human cases from Serbia are available for the studied period. However, collected DOBV isolates in this work phylogenetically clustered together with isolates from Serbian human cases dating from 2002, with 1.9% nucleotide divergence. We determined the level of kinship between seropositive and seronegative animal groups and found no significant difference, suggesting that horizontal virus transmission in the studied population was the same within and among the hatches. Our findings are the first genetic detection of DOBV in rodents in Serbia. We confirm wide and continuous hantavirus presence in the examined parts of the Balkans, underlying the necessity of continual monitoring of hantavirus circulation in A. flavicollis.

B chromosomes in populations of mammals.

B chromosomes (Bs) have been found in 55 out of 4629 living species of mammals. The summarized data show great variability in types of mammalian Bs, including differences in size, shape and molecular composition. This variability extends to the origin, mode of transmission and population dynamics. In general, B chromosomes in mammals do not differ from Bs found in other animal or plant species, but some peculiarities do exist. Most species in which Bs are found are widespread. Some data support the view that Bs may contribute to the successful expansion of some of these species, but it is possible that Bs are just more easily scored in them due to their frequent occurrence. Most of these species are also characterized by cycling fluctuations of abundance and characteristic social organization that produce conditions favorable for Bs to spread. All areas of research on Bs in mammals suffer from lack of data, emphasizing the necessity for intensified research on the molecular structure and ways of maintenance of Bs in populations.

Antimutagenic effects of extracts from sage (Salvia officinalis) in mammalian system in vivo.

Naturally occurring antimutagenic factors, especially those of plant origin, have recently become a subject of intensive research. Antimutagenic properties of terpenoid fractions of sage (Salvia officinalis) were tested in mammalian system in vivo through examining the ability of sage to decrease the frequency of aberrant cells induced by a potent mutagen. First, groups of mice were treated with three concentrations of sage alone and it was established that the frequency of aberrant cells after treatment with a concentration of 25 microL/kg was not significantly different from the negative control (olive oil), while that found after treatment with the 50 microL/kg concentration differed significantly (chi2(1) = 4.05, p < 0.05). Sage used at a concentration of 100 microL/kg was cytotoxic. Mitomycin C (MMC), known as a potent mutagen, was used for induction of chromosome aberrations. Post-treatment with sage suppressed the effects of MMC significantly. Both concentrations (25 microL/kg and 50 microL/kg) produced a significant decrease in the frequency of aberrations relative to MMC (chi2(1) = 5.42, p < 0.02, chi2(1) = 14.93, p < 0.001, respectively). The percent of aberrations decreased with increasing concentrations of sage. Only nontoxic concentrations of sage without mutagenic effects can be recommended for use as inhibitors of mutagenesis or carcinogenesis.

[Transmissible spongiform encephalopathies]. / Transmisivne spongioformne encefalopatije.

The great interest for transmissible spongiform encephalopathies, transmissible neurodegenerative diseases has been rapidly grown in the last 10 years, especially after their epidemic appearance in domestic and wild animals in Great Britain and the Bovine Spongiform Encephalopathy ("mad cow diseases") in England has provoked the increased interest in veterinary and medical researches. The opinion that prions are due to transmissible spongiform encephalopathies was accepted by Prusiner and al. 1982. Prions are defined as "Little infective pathogens, which contain protein and which are resistant to procedures which modify and hydrolyze nucleic acid". These diseases in animals and humans have a long incubation period and a long clinical course, and are always fatal. Neuropathological features consist of neuronal vacuolization, neuronal death and gliosis with hyperastrocytosis. Transmissible neurodegenerative diseases are transmissible, infectious and inherited diseases. The inflammatory syndrome, immunological reactions or interferon cannot be detected in the central nervous system. The precise diagnosis of transmissible neurodegenerative diseases can be established only by the examination of the central nervous system after biopsy or autopsy. During the last two years a new variation of Creutzfeld-Jakob disease was detected in England which is different from the classical Creutzfeld-Jakob diseases. The clinical course and the absence of EEG features are typical for Creutzfeld-Jakob disease, and neuropathological changes show extensive curu plaques in the cerebrum and the cerebellum. According to the latest researches it has been established that the abnormal PrP, PrPsc are the same or similar strains which cause the bovine spongiform encephalopathy and a new variation of Creutzfeld-Jakob disease, what is of great medical and epidemiological significance.

[The role of Epstein-Barr virus infections in human pathology]. / Uloga Epstein-Barrove virusne infekcije u humanoj patologiji.

Epstein-Barr virus (EBV) belongs to the group of herpesvirus and may remain latent after primary infection with a possibility of periodical reactivation EBV infection is transmitted by intimate, oral contact with previously infected persons, persons sick or infected earlier which secrete virus periodically. We report on pathogenesis of primary infection as well as clinical characteristics of infective mononucleosis and other EBV associated diseases (Birkitt lymphoma, nasopharyngeal carcinoma, syndrome of chronical fatigue).

B chromosomes and developmental homeostasis in the yellow-necked mouse, Apodemus flavicollis (Rodentia, Mammalia): effects on nonmetric traits.

B chromosomes are found in almost all populations of the yellow-necked mouse, Apodemus flavicollis (Rodentia, Mammalia). Their effects on developmental homeostasis in this species were analyzed using morphological nonmetric traits (number of foramina) in a sample of 218 animals from locality Mt Jastrebac in the former Yugoslavia. Variations of the parameters of developmental homeostasis (the degree of fluctuating asymmetry--FA, the number of asymmetrical characters per individual--NA, and the total phenotypic variability--PV) were examined in three groups: in animals without Bs, with one B chromosome, and with more than one B chromosome. Significant differences in the level of FA between groups of animals were found for two characters. Carriers of one B chromosome displayed the highest level of phenotypic variability. Parameters of developmental stability (DS) were monitored in the population in which significant variations in the frequency of animals with Bs (fB) were established during the season earlier. The FA levels for four foramina out of a total of 12 examined followed the changes of frequencies of animals with Bs. Furthermore, a significant seasonal correlation between NA and fB was found. The presence of B does not cause a disturbance of homeostasis in a way that allows changes in homeostasis to be directly related to B chromosome's presence. However, carriers of B react differently to environmental changes than do noncarriers.

The role of B chromosomes in the population dynamics of yellow-necked wood mice Apodemus flavicollis (Rodentia, Mammalia).

Seasonal changes in the frequency of animals with B chromosomes were studied during 1989 in the population of yellow-necked wood mice, Apodemus flavicollis, on the mountain Jastrebac in Yugoslavia. During the study the estimated density of animals reached a very large value (378.8 individuals/ha for the April sample), which produced stress conditions leading to population decline. Six samples with a total of 247 specimens were karyotypically analyzed. Animals with Bs were present in all samples in frequencies that differed significantly at the 5% level. This difference was produced by the reduction in frequencies of specimens with Bs between the April and May samples (0.53 versus 0.29; chi 2(1) = 9.44; p < 0.002) and appeared mostly as a result of increased elimination of B-carrying animals from the prereproductive group. Young animals with Bs were inferior in stress conditions produced by overabundance of animals. From the first to the last sample, the frequency of animals with one B among B-containing animals constantly decreased, while the frequency of animals with more than one B increased. The possible role of Bs in regulating population dynamics is discussed.

[Intrathecal synthesis of immunoglobulins during acute viral meningitis with high cerebrospinal fluid protein levels]. / Intratekalna sinteza imunoglobulina u toku akutnih virusnih meningitisa sa hiperproteinorahijom.

The article deals with the results of examination of immunoglobulins (IgM, IgA, IgG) in the liquor and serum of 50 patients with acute viral meningitis and hyperproterinorashia over 1.0 g/l. The quantitative determination of immunoglobulins was done by the method of radial immunodiffusion according to Manzoni. The confirmation of intrathecal synthesis of immunoglobulins was done by liquor indexes. IGM intrathecal synthesis was found in 11 (22%) patients, IgA in 16 (32%) and IgG in 10 (20%) subjects. The daily amount of IgG synthesis in the central nervous system was 1.65-97.15 mg (mean 26.60 mg). The values of liquor indexes were decreased in later disease. This suggested the decrease of disappearance of immunoglobulin intrathecal synthesis. There were 26 control patients with meningism (meningeal syndrome with regular cytobiochemical finding in the liquor). IgG intrathecal synthesis was established in one patient with meningism. This finding was an extremely rare and unusual finding.

[Neurolues--clinical characteristics, diagnosis and therapy]. / Neurolues--klinicke karakteristike, dijagnoza i terapija.

A female patient with neurolues was presented in this paper. The disease started 25 years after primoinfection by neurologic disturbances which corresponded to classic neurolues with the signs of meningovascular and parenchymatous disease. The diagnosis was confirmed by positive serologic tests, computerized brain tomography--whereby syphilomas were found in the brain and by histopathologic survey of one of peripheral syphilomas. Antibiotic therapy with high penicillin doses was initiated but without more significant clinical effects, it only led to negativization of serologic tests. The patient died after two months. Serologic tests to syphilis should be temporarily repeated in all patients suspective of neurolues, in order to initiate or repeat the previously employed therapy.

The protein composition of the hepatocyte nuclear matrix is differentiation-stage specific.

The protein composition of hepatocyte nuclear matrices was examined in rats from the 16th day of gestation to 75 days after birth (adult). An overall increase in size of the nuclear matrix was accompanied by quantitative and qualitative changes in its protein content. Quantitative changes of the major proteins of the peripheral lamina surrounding the isolated nuclear matrix were detected. By Western analysis we established that in pre- and postnatal nuclear matrices the relative concentrations of lamin C were greater than lamin A. After birth, the relative concentrations of both lamins progressively increased. In the adult nuclear matrix, the concentration of lamin A was greater than lamin C. In contrast, the relative concentrations of lamin B remained unchanged throughout development and growth. The relative concentrations of two nuclear matrix-associated regulatory proteins studied changed with development and growth: transcription factor C/EBPalpha isoforms, which were detected during the gestation period, increased notably after the first postnatal day, attaining a maximum at the adult stage; the high concentrations of the proliferating cell nuclear antigen (PCNA) perceptibly decreased after the 21st prenatal day. Changes in the composition of the nuclear matrix protein suggest that this structure coordinates nuclear functioning during cell differentiation.

DNA profiling of B chromosomes from the yellow-necked mouse Apodemus flavicollis (Rodentia, Mammalia).

Using AP-PCR-based DNA profiling we examined some structural features of B chromosomes from yellow-necked mice Apodemus flavicollis. Mice harboring one, two, or three or lacking B chromosomes were examined. Chromosomal structure was scanned for variant bands by using a series of arbitrary primers and from these, informative bands were selected. The selection criteria used were the ability to differentiate between individuals of the species, to detect markers common for both A and B chromosomes, and, importantly, to differentiate between A- and B-chromosome sets. In addition to primers, profiling conditions were found to be critical for meeting the selection criteria. Primers and analysis conditions that demonstrated structural characteristics unique to the B-chromosome set are described. These characteristics included variant bands as qualitative parameters and altered electrophoretic band intensities as quantitative distinctions estimated by integration of densitometric profiles of electrophoretograms. B chromosome-specific molecular markers are easy to detect by AP-PCR-based DNA profiling in the presence of a full set of A chromosomes. Models for the origin of yellow-necked mouse B chromosomes are discussed in the context of presented data.

[Acute meningitis without cutaneous manifestations caused by varicella zoster virus--intrathecal synthesis of specific antibodies]. / Akutni meningitis izazvan varicella zoster virusom bez kutanih manifestacija--intratekalna sinteza specificnih antitela.

The results of virusologic and cytobiochemical evaluation of CSF and serum samples of four patients with acute viral meningitis (AVM), most probably induced by varicella zoster virus (VZV), are reported. In no case VZV infection was not cutaneously manifested. Aetiologic diagnosis was established according to the presence of specific anti VZV AVM, in spite of their presence in the sera. On samples detected by indirect enzyme immunoassay (EIA). Four different antibody indexes were used to prove that the antibodies were intrathecally synthesized. Other viral antibodies (HSV, mumps) were not evident in the CSF samples of the patients with VZV avm, in spite of their presence in the sera. On the other hand, anti VZV antibodies could not be identified in the CSF samples of the controls (AVM of other aetiology, meningism) in spite of their presence in the sera. A possible aetiologic link between anti VZV antibodies presence in the CSF samples and some neurologic syndromes is discussed.

[Significance of bromide levels in evaluation of the hemato-encephalic barrier during acute viral meningitis]. / Znacaj bromidnog kolicnika za procenu funkcionog stanja hematoencefalne barijere u toku akutnih virusnih meningitisa.

INTRODUCTION: Inflammatory diseases of the CNS are associated with increased permeability of blood-brain barrier due to vasculitis of cerebral blood vessels leading to regional ischaemia and necrosis. Many substances pass easily from the serum into the CSF through inflamed and damaged blood-brain barrier. Bromide partition test is one of the parameters for evaluation of the blood-brain barrier integrity during CNS inflammatory diseases. As early as 1929 Walter published a monograph on blood-brain barrier accentuating the value of bromide partition test for evaluation of the functional status of blood-brain barrier. In healthy individuals bromide partition test usually ranges from 2.9-3.5. In most patients with TB meningitis bromide partition test remains below 1.6 while it is usually higher in patients with serous meningitis of other aetiologies. However, the low bromide partition test may associate also some other viral and other serous meningitides accompanied with severe lesions of the blood-brain barrier, resulting in increased passage of sodium bromide from the serum into the CSF, so that the serum/CSF ratio is lower. The aim of th study was to calculate and analyze the results of bromide partition test in patients with acute viral meningitis and hyperproteinorachia above 0.70 g/l and to calculate and analyze the results of bromide partition test in patients with TB and parasitic meningitis. The results of bromide patirion tests were compared with results of other parameters for evaluation of the blood-brain barrier function. METHOD: Titration method was used to determine the serum sodium bromide and CSF ratio. Sodium bromide oxidation into sodium bromate was the principle, where addition of potassium iodide is followed by release of equivalent amount of elementary iodide. Iodide is titrimetrically evidenced with thyosulphate, with starch as an indicator. In addition to bromide partition test functional condition of blood-brain barrier was also evaluated by albumin coefficients, albuminorachia and total proteinorachia. RESULTS: In 6 of 30 patients (20%) with acute viral meningitis the bromide partition ratio was below 1.6, suggesting severe damage of blood-brain barrier in these patients: albumin coefficients exceeded 16.7 (mean = 24.7), albuminorachia was over 0.90 (mean = 1.38 g/l), and total proteinorachia over 3.0 g (mean = 3.1 g/l). The aetiology was confirmed in all patients: lymphocyte choriomeningitis virus in 5, COX B1 in one. Values of bromide partition test were higher on follow-up examination, and values of other parameters decreased, suggesting restitution of blood-brain barrier. The results are given in Table 1. Most severe damage of blood-brain barrier was evidenced in patients with lymphocyte choriomeningitis. Parameters for evaluation of functional condition of blood-brain barrier in patients with lymphocyte choriomeningitis and TB meningitis with most severe damage of the barrier, were compared. The results are given in Table 2. No differences between the compared parameters were found, i.e. no difference in the severity of blood-brain damage. Bromide patition test values were below 1.6 in all patients with TB meningitis and in 3 of 5 patients with parasitic meningitis. DISCUSSION: Numerous studies have shown that bromide patition test value during TB meningitis was usually below 1.6. This is probably due to hypersensitivity reaction of the meninges to tuberculin which is located intrathecally and leads to blood-brain barrier damage. Use of antituberculous drugs does not affect the bromide partition test values in early stages of the disease. In all of our 15 patients with TB meningitis the bromide partition test values were below 1.6, coinciding with reference reports. However, we also measured bromide partition test values below 1.6 in 6 (20%) of 30 patients with acute viral meningitis with proteinorachia above 0,70 g/l (ABSTRACT TRUNCATED).

[Clinical expirience in the treatment of purulent meningitis and sepsis with Ceftriaxon]. / Klinicka iskustva sa primenom cefriaksona u lecenju gnojnih menigitisa u sepsi.

Our expirience in the treatment of purulent meningitis and sepsis with third generation of cephalosporin--Cephtriaxon--are presented. In the last two years 13 of 44 patients suffering from purulent meningitis were treated with Cephtriaxon. They were all succesfuly cured without complications and recurrence. The time of CSF normalisation was from 8-17 days, depending on the aethyological agent of meningitis. Cephtriaxon is the drug of choice for E. coli and Hemophilus influence meningitis. We achiewed good results in the treatment ob sepsis, and ten of these cases are presented. Our Staphylococcus aureus showed good susceptibility to Cephtriaxon, which is the reason of the good therapeutical results. The analysis of antibacterial activity of Cephtriaxon in bacteria isolated from blood and CSF cultures, revealed that 80 per cent of isolated bacteria were highly susceptible.

[Cerebellitis during Mycoplasma pneumoniae infection]. / Cerebelitis u toku infekcije Mycoplasmom pneumoniae.

Mycoplasma pneumoniae is the primary cause of respiratory system diseases in school children and young adults. Most often an infection of the upper respiratory airways is in question, with or without pharyngitis, while clinically manifest pneumonia occurs in only 3-10% of patients. However, multisystemic manifestations of Mycoplasma pneumoniae infections can also occur, usually during the first 14 days after the start of respiratory symptomatology, but can also occur as isolated in some cases, with no prior respiratory illness, nor its later occurrence. In our patient there came to a development of pneumonia, exudative pleuritis, sinusitis and cerebellitis. The duration of the disease was protracted, but recovery was complete, without sequelae. The etiological diagnosis was set retrospectively on the basis of the dynamics of the titer of complement fixating antibodies. Mycoplasma pneumoniae should be considered as a possible etiological agent of various clinical syndromes.