Matrix bound phosphine formation and depletion in eutrophic lake sediment fermentation-simulation of different environmental factors

Closed anaerobic batch-fermentation of eutrophic lake sediment samples was performed under variation of four environmental fermentation factors (pH, temperature, water/sediment ratio and disturbance) to learn how the quantity of phosphine will change and if the quantity of phosphine can increase. The fermentation conditions where matrix bound phosphine (MBP) increased (doubled from 3193+/-520 to about 7982+/-1003 ng/kg) were: a pH of 8 and of 10 (as compared to 1, 2, 4, 6, 12), a temperature of 20 and 30 degrees C (as compared to 4 and 40 degrees C), a water/sediment ratio of 3:1 (as compared to 1:1, 2:1, 5:1) and a disturbance of 100 r/min (as compared to 0 r/min), respectively. Although, over the full time course of fermentation, the balance of phosphine production became negative again or did remain almost unchanged under most conditions. A pH of 1 or disturbance of 150 r/min was significant factors to decrease phosphine over the long term. Free phosphine had been detected but was of minor importance (in the order of 60.9+/-10.1 ng/m(3)). Overall, the fermentation conditions which had been most favorable for microbial life (moderate temperature (20 and 30 degrees C) and pH 8) were also most favorable for a positive phosphine balance. This is an indication, but no biochemical proof that a natural (biogenic, microbial, biochemical) NET PRODUCTION of phosphine or DE NOVO PRODUCTION of phosphine has occurred. MBP concentrations in lake sediments were discussed as to be strongly dependent on a balance of natural generation and depletion processes, dependent of the simulated parameters.     

New Fluorescent Bichromophoric Probes for the Donor–Donor Energy Migration (DDEM) Studies

Three fluorescent probes were synthesized for studying the excitation energy migration between two identical fluorophores. Each probe has two identical fluorescent groups (dansyl, 7-nitrobenzo-2-oxa-1,3-diazole-4-yl, or fluoresceinyl) linked by the rigid bis-(8-aminooctyl)amide of 4,4"-biphenyldicarbonic acid or flexible dotriacontanedioic acid spacer, which enables the intramolecular energy migration through the distance of 3.2–3.5 nm.     

Surveillance of Fish Diseases in the Nordic Countries

Due to the increasing importance of disease problems in the fish farming industry and the impact disease may have on both feral and farmed fish in the Nordic countries, monitoring and surveillance on diseases have for many years been considered to be of socioeconomic importance. All the Nordic countries have a national legislation as basis for their surveillance and disease control in aquatic animals and regulations listing notifiable diseases of concern to the countries. The list of diseases vary between the countries. In addition, Denmark, Finland and Sweden are ruled by [3] as regards placing on the market of aquaculture animals and products. The surveillance for viral diseases in all the Nordic countries has mainly been based on the testing procedures given in the EU Commission [2].     

Cell-free derivatives from mesenchymal stem cells are effective in wound therapy

AIM:To compare the efficacy of cell-free derivatives from Bone marrow derived human mesenchymal stem cells(hMSCs) in wound therapy.METHODS:hMSCs have been shown to play an important role in wound therapy.The present study sought to compare efficacy of hMSCs and cell-free derivatives of hMSCs,which may be clinically more relevant as they are easier to prepare,formulate and transport.hMSCs were isolated from human bone marrow and cultured.Multi lineage differentiation of hMSCs was performed to confirm their identity.The ability of hMSCs to migrate was evaluated using in vitro and in vivo migration assays.Cell lysates and conditioned medium concentrate was prepared from hMSCs(see Methods for details).Wounds were induced in mice and wound areas were measure before and after cell and cell-free derivative treatment.RNA and proteins were extracted from the skin and cytokine levels were measured.RESULTS:Co-culture of hMSCs with keratinocytes resulted in increased expression of CXCL-12(SDF1) and ENA78(CXCL-5) in the conditioned media indicating that the hMSCs can respond to signals from keratinocytes.Accelerated wound closure was observed when hMSCs were injected near the site of excisional wounds in athymic as well as NOD/SCID mice.Interestingly,cell-free lysates prepared from hMSCs were also effective in inducing accelerated wound closure and increased expression of SDF1 and CXCL-5 at the wound bed.Additionally,concentrated media from hMSCs as well as an emulsion containing lysates prepared from hMSCs was also found to be more effective in rapid re-epithelialization than fibroblasts or vehicle-alone control.Use of cell-free derivatives may help replace expensive wound care approaches including use of growth factors,epidermal/dermal substitutes,synthetic membranes,cytokines,and matrix components,and most importantly avoid transmission of pathogens from human and animal products.CONCLUSION:These results encourage development of derivatives of hMSCs for wound care and re-epithelialization applications.     

Seroprevalence of Taenia solium infections in Croatian patients presenting with epilepsy

Epilepsy is one of the most common neurological disorders, while neurocysticercosis caused by Taenia solium infection of the central nervous system currently represents the leading cause of secondary epilepsy in Central and South America, East and South Asia, and sub-Saharan Africa. As a result of increased migration from these endemic regions, neurocysticercosis and subsequent epilepsy are becoming a growing public health problem in developed countries as well. In order to determine the prevalence of T. solium infection in patients with epilepsy in Croatia, a retrospective serological study was conducted. A total of 770 serum samples were tested for the presence of T. solium IgG antibodies using a commercial qualitative enzyme immunoassay. The Western blot technique was used as a confirmatory test for the diagnosis. The overall seroprevalence rate of T. solium infection in patients with clinically proven epilepsy was 1.5%. Although the results have shown that infection with this tapeworm is rare in Croatia, this study hopes to increase awareness about the importance of preventive measures and benefits of accurate and timely diagnosis. Intervention measures for infection control are crucial, namely sanitation improvement, control of domestic pig-breeding, detailed meat inspection, detection and treatment of tapeworm carriers, hand washing and health education.     

Are Patients with Erythema Migrans Who Have Leukopenia and/or Thrombocytopenia Coinfected with Anaplasma phagocytophilum or Tick-Borne Encephalitis Virus?

Lyme borreliosis (LB), tick-borne encephalitis (TBE) and human granulocytic anaplasmosis (HGA) are endemic in central part of Slovenia. We tested the hypothesis that patients with erythema migrans (EM) from this region, who have leukopenia and/or thrombocytopenia (typical findings in HGA and in the initial phase of TBE but not in patients with LB) are coinfected with Anaplasma phagocytophilum and/or with TBE virus, i.e. that cytopenia is a result of concomitant HGA or the initial phase of TBE. Comparison of clinical and laboratory findings for 67 patients with EM who disclosed leukopenia/thrombocytopenia with the corresponding results in sex- and age-matched patients with EM and normal blood cell counts revealed no differences. In addition, patients with typical EM and leukopenia and/or thrombocytopenia tested negative for the presence of IgM and IgG antibodies to TBE virus by ELISA as well as for the presence of specific IgG antibodies to A. phagocytophilum antigens by IFA in acute and convalescent serum samples. Thus, none of 67 patients (95% CI: 0 to 5.3%) with typical EM (the presence of this skin lesion attests for early Lyme borreliosis and is the evidence for a recent tick bite) was found to be coinfected with A. phagocytophilum or had a recent primary infection with TBE virus. The findings in the present study indicate that in Slovenia, and probably in other European countries endemic for LB, TBE and HGA, patients with early LB are rarely coinfected with the other tick-transmitted agents.     

Tick-Borne Encephalitis Virus Infects Rat Astrocytes but Does Not Affect Their Viability

Tick-borne encephalitis virus (TBEV) causes one of the most dangerous human neuroinfections in Europe and Asia. To infect neurons it must cross the blood-brain-barrier (BBB), and presumably also cells adjacent to the BBB, such as astrocytes, the most abundant glial cell type. However, the knowledge about the viral infection of glial cells is fragmental. Here we studied whether TBEV infects rat astrocytes. Rats belong to an animal group serving as a TBEV amplifying host. We employed high resolution quantitative fluorescence microscopy to investigate cell entry and cytoplasmic mobility of TBEV particles along with the effect on the cell cytoskeleton and cell survival. We report that infection of astrocytes with TBEV increases with time of exposure to TBEV and that with post-infection time TBEV particles gained higher mobility. After several days of infection actin cytoskeleton was affected, but cell survival was unchanged, indicating that rat astrocytes resist TBEV-mediated cell death, as reported for other mammalian cells. Therefore, astrocytes may present an important pool of dormant TBEV infections and a new target for therapeutic intervention.     

Cilia are required for asymmetric <Emphasis Type="Italic">nodal</Emphasis> induction in the sea urchin embryo

Background

Left-right (LR) organ asymmetries are a common feature of metazoan animals. In many cases, laterality is established by a conserved asymmetric Nodal signaling cascade during embryogenesis. In most vertebrates, asymmetric nodal induction results from a cilia-driven leftward fluid flow at the left-right organizer (LRO), a ciliated epithelium present during gastrula/neurula stages. Conservation of LRO and flow beyond the vertebrates has not been reported yet.

Results

Here we study sea urchin embryos, which use nodal to establish larval LR asymmetry as well. Cilia were found in the archenteron of embryos undergoing gastrulation. Expression of foxj1 and dnah9 suggested that archenteron cilia were motile. Cilia were polarized to the posterior pole of cells, a prerequisite of directed flow. High-speed videography revealed rotating cilia in the archenteron slightly before asymmetric nodal induction. Removal of cilia through brief high salt treatments resulted in aberrant patterns of nodal expression. Our data demonstrate that cilia - like in vertebrates - are required for asymmetric nodal induction in sea urchin embryos.

Conclusions

Based on these results we argue that the anterior archenteron represents a bona fide LRO and propose that cilia-based symmetry breakage is a synapomorphy of the deuterostomes.