Unexpected genetic diversity of Fallopia japonica from Central Europe revealed after AFLP analysis

Recently much attention has been paid to genetic aspects of invasive success in Japanese knotweed s.l. One hypothesis to explain the invasive spread of these species is a multiple introduction, which leads to a higher level of genetic diversity in the invaded range. Fallopia japonica is considered to be genetically uniform in Europe, introduced as a single female clone. However, there is some evidence suggesting that invasion history and dynamics differ between Western and Central-Eastern Europe. We used AFLP markers to characterize genetic diversity of three Fallopia taxa that occur in Poland: F. japonica, F. sachalinensis and their hybrid Fallopia × bohemica, growing in so-called ‘homogeneous’ populations, consisting of one taxon and ‘heterogeneous’ populations, composed of the three taxa cohabiting together. No polymorphism, resp. an insignificantly low variability was observed in the ‘homogeneous’ populations. In the ‘heterogeneous’ stands polymorphism was detected within each taxa, with one exception that concerns individuals of F. sachalinensis from a riparian habitat. The highest level of polymorphism was found among individuals of F. × bohemica. The most striking result of our study is the observation of polymorphism between individuals of F. japonica. The AFLP data also showed that F. × bohemica is most diverse when occurring in a heterogeneous configuration with F. japonica and F. sachalinensis in the same habitat. Our results are the first evidence of genetic diversity in F. japonica populations in Central Europe and can implicate the possibility of its multiple introduction in this region or the existence of sexual reproduction of this species.     

A New Real-Time PCR for the Detection of Plasmodium ovale wallikeri

It has been proposed that ovale malaria in humans is caused by two closely related but distinct species of malaria parasites: P. ovale curtisi and P. ovale wallikeri. We have extended and optimized a Real-time PCR assay targeting the parasite’s small subunit ribosomal RNA (ssrRNA) gene to detect both these species. When the assay was applied to 31 archival blood samples from patients diagnosed with P. ovale, it was found that the infection in 20 was due to P. ovale curtisi and in the remaining 11 to P. ovale wallikeri. Thus, this assay provides a useful tool that can be applied to epidemiological investigations of the two newly recognized distinct P. ovale species, that might reveal if these species also differ in their clinical manifestation, drugs susceptibility and relapse periodicity. The results presented confirm that P. ovale wallikeri is not confined to Southeast Asia, since the majority of the patients analyzed in this study had acquired their P. ovale infection in African countries, mostly situated in West Africa.     

Heteromers of Glutamate Decarboxylase Isoforms Occur in Rat Cerebellum

Abstract: The subunit structure of brain glutamate decarboxylase in cerebellum was investigated by using gel electrophoresis and antisera that specifically recognize the individual isoforms of brain glutamate decarboxylase (termed GAD₆₅ and GAD₆₇). The antisera were prepared against peptides that corresponded to amino acid sequences specific to each isoform. Each antiserum reacted specifically with the appropriate peptide in an ELISA and with the appropriate form of GAD on immunoblots. Nondenaturing gradient gel electrophoresis indicated that GAD is principally multimeric with monomeric forms comprising <3% of the total. Immunoprecipitation and immunoaffinity chromatography experiments were performed with antisera W624 and W883, which were prepared against peptides specific to GAD₆₅ and GAD₆₇, respectively. Immunoprecipitates prepared from cerebellar supernatants with W624 contained both GAD₆₅ and GAD₆₇, whereas some GAD₆₇ was left in the supernatant. In a similar manner, immunoprecipitates prepared with W883 contained both GAD₆₅ and GAD₆₇, whereas some GAD₆₅ remained in the supernatant. In addition, immunoaffinity columns prepared with either W624 or W883 retained both GAD₆₅ and GAD₆₇ even after extensive washing. These results are consistent with the presence of heteromultimers of GAD₆₅ and GAD₆₇ in cerebellum in addition to homomers of each form.     

High Frequency Ultrasound for In Vivo Pregnancy Diagnosis and Staging of Placental and Fetal Development in Mice

Background

Ultrasound is a valuable non-invasive tool used in obstetrics and gynecology to monitor the growth and well being of the human fetus. The laboratory mouse has recently emerged as an appropriate model for fetal and perinatal studies because morphogenetic processes in mice exhibit adequate homology to those in humans, and genetic manipulations are relatively simple to perform in mice. High-frequency ultrasound (HFUS) has recently become available for small animal preclinical imaging and can be used to study pregnancy and development in the mouse. The objective of the current study was to assess the main applications of HFUS in the evaluation of fetal growth and placental function and to better understand human congenital diseases.

Methodology/Principal Findings

On each gestational day, at least 5 dams were monitored with HFUS; a total of ∼200 embryos were examined. Because it is not possible to measure each variable for the entire duration of the pregnancy, the parameters were divided into three groups as a function of the time at which they were measured. Univariate analysis of the relationship between each measurement and the embryonic day was performed using Spearman’s rank correlation (Rs). Continuous linear regression was adopted for multivariate analysis of significant parameters. All statistical tests were two-sided, and a p value of 0.05 was considered statistically significant.

Conclusions/Significance

The study describes the main applications of HFUS to assess changes in phenotypic parameters in the developing CD1 mouse embryo and fetus during pregnancy and to evaluating physiological fetal and placental growth and the development of principal organs such as the heart, kidney, liver, brain and eyes in the embryonic mouse. A database of normal structural and functional parameters of mouse development will provide a useful tool for the better understanding of morphogenetic and cardiovascular anomalies in transgenic and mutant mouse models.     

Structure based molecular design, synthesis and biological evaluation of α-pyrone analogs as anti-HSV agent

Several options for treating Herpes Simplex Virus type 1 and type 2 are available. However, non-specific inhibition and drug resistance warrants the discovery of new anti-herpetic compounds with better therapeutic profile or different mode of action. The non-nucleoside inhibitors of HSV DNA polymerase target the site that is less important for the binding of a natural nucleoside or nucleoside inhibitors. In the present study, we have explored the possibility to find a new lead molecule based on α-pyrone analogs as non-nucleoside inhibitors using structure based modeling approach. The designed molecules were synthesized and evaluated for anti-HSV activity using MTT assay. The compound 5h with EC(50) 7.4μg/ml and CC(50) 52.5μg/ml was moderately active against HSV when compared to acyclovir. A plaque reduction assay was also carried out and results reveal that 5h is more effective against HSV-1 with better selective index of 12.8 than against HSV-2 (SI=3.6). The synthesized compounds were also evaluated for anti-HIV activity, but none were active.