Glycogen breakdown in cleaving Xenopus embryos is limited by ADP.

Xenopus eggs contain large stores of glycogen, but this glycogen is not glycolytically processed during cleavage. The Embden-Meyerhof pathway is inhibited by the absence of pyruvate kinase activity in vivo, and lactate and pyruvate are present at relatively low levels. In the late blastula, just preceding gastrulation, lactate levels increase, indicating the onset of glycogen breakdown and glycolytic flux. Glycolysis from microinjected [14C]glucose-6-phosphate could be transiently activated, however, by the coinjection of ADP into fertilized eggs, and constitutively activated by the injection of the ATPase potato apyrase, indicating the presence of all enzymes necessary for glycolytic activity. The isozyme profiles of pyruvate kinase and malic enzyme, two enzymes involved in carbon metabolism during cleavage or in the subsequent activation of glycogen breakdown, do not change between the egg and gastrula stages. These data suggest that the activation of glycogen breakdown and glycolysis in the late blastula is probably not a result of new gene activity but may be the metabolic consequence of increased free ADP that is then able to support the pyruvate kinase reaction.     

Plasma copeptin in the assessment of febrile neutropenia

Copeptin, the surrogate marker of arginine vasopressin (AVP), has been suggested to be a useful biomarker in monitoring sepsis reflecting hemodynamic imbalance and stress state. This prospective study conducted at a hematology ward in a Finnish University Hospital aimed to investigate whether plasma copeptin predicts the development of complicated course of neutropenic fever (bacteremia or need for treatment at intensive care unit) in 100 hematological patients experiencing their first neutropenic fever episode after intensive chemotherapy for hematological malignancy. Contrary to study presumptions, not elevated copeptin but the lack of a proper initial increase of plasma copeptin (<0.02 ng/mL from day 0 to day 1) predicted blood culture positive sepsis (p=0.023) and gram-negative bacteremia (p=0.045). No correlation was observed with plasma sodium, blood pressure or evaluated osmolality. Plasma copeptin correlated inversely with the same day pentraxin 3 on day 0-day 2 (all p-values <0.001) and with C-reactive protein on day 1 (p=0.015). In conclusion, copeptin did not correlate with disease severity, but the lack of a proper initial increase was associated with bacteremic complications of febrile neutropenia in hematological patients. The findings suggest the possibility of central dysregulation of AVP release and do not support the use of copeptin as a biomarker of septic complications in this patient group.     

Stimulation of Epstein-Barr virus-infected human B cell growth by physiological concentrations of 4-hydroxynonenal

We had previously shown that cyclosporin A (CsA) directly promoted the immortalization of Epstein-Barr virus (EBV)-infected human B cells (EBV-B cells) via an oxidative stress mechanism. 4-Hydroxynonenal (HNE) is a reactive end-product of lipid peroxidation. We hypothesized that HNE may mediate a direct oxidative stress-promoting effect of CsA on EBV-B cells. HNE-protein adducts in CsA-treated EBV-B cell extracts were assayed immunochemically using a Slot-Blot method. Cell proliferation was assayed by [(3)H]-thymidine incorporation. EBV oncogene latent membrane protein-1 (LMP1) expression was assayed by using PE-conjugated anti-LMP1 antibody in flow cytometry. We found that CsA at 500 ng ml(-1) and 1000 ng ml(-1) significantly increased the level of HNE-protein adducts in EBV-B cells over the control (arbitrary units +/- SE) by 251.3 +/- 7.5 to 361.3 +/- 9.7 and 342.7 +/- 10.7, respectively (p < 0.05, n = 3). EBV-B cells treated with a physiological concentration of HNE (1 microM) for 0.5 and 1 h and cultured for 2 and 4 weeks showed significantly increased [(3)H]-thymidine incorporation. EBV-B cells treated with HNE (1 microM) for 1 h and subsequently cultured for 2 and 4 weeks had a significantly higher ( > 2.0 times) LMP1-positive cell population over the control. In conclusion, in accordance with our previous findings, we show that CsA treatment of EBV-B cells results in increased production of the lipid peroxidation reactive end-product HNE that directly promotes EBV-B cell proliferation and LMP1 expression. This observation provides evidence for further understanding the mechanism of CsA-induced oxidative stress on EBV-related post-transplant lymphoproliferative disorder (PTLD).     

Seedling establishment of Asteraceae forbs along altitudinal gradients: a comparison of transplant experiments in the native and introduced ranges

Aim Since ecological and evolutionary context changes when a plant species is introduced to a new area, it can be assumed that responses of alien plants to changing conditions along environmental gradients differ from those in their native range. Even if seed availability is not limited, the distribution of alien plants along such a gradient might still be restricted by their ability to germinate and establish as seedlings. In the present study, we aim at testing what factors promote or limit plant invasions during early establishment by using altitude as a model gradient. Location Altitudinal gradients in the Wallowa Mountains (Oregon, USA) and the Swiss Alps (Valais, Switzerland). Methods In transplant experiments along altitudinal gradients, we investigated the early establishment success of eight invasive alien Asteraceae species in their native and introduced ranges in the Wallowa Mountains and the Swiss Alps. Results Seedling recruitment was not restricted to relatively lower altitudes in the introduced range. In addition, we found no evidence for genetic adaptation along the altitudinal gradient in the introduced range, highlighting the importance of phenotypic flexibility for invasions. Furthermore, seedling recruitment was only enhanced by disturbance in the native range where vegetation was comparably dense but not in the introduced range. However, plant development was strongly delayed in the introduced range, probably due to low seasonal water availability. Main Conclusions We conclude that introduced plants, due to their ability to tolerate a wide range of environmental conditions, are not necessarily more restricted in their altitudinal limits than they are in their native range. Furthermore, due to other interacting factors (e.g. different competition situations among ranges), attempts to predict distributions of alien plants in the introduced range that are based on their distributions in the native range may be misleading.     

Large-scale in vitro embryo production and pregnancy rates from Bos taurus, Bos indicus, and indicus-taurus dairy cows using sexed sperm

Herein we describe a large-scale commercial program for in vitro production of embryos from dairy Bos taurus, Bos indicus, and indicus-taurus donors, using sexed sperm. From 5,407 OPU, we compared the number of recovered oocytes (n = 90,086), viable oocytes (n = 64,826), and embryos produced in vitro from Gir (Bos indicus, n = 617), Holstein (Bos taurus, n = 180), 1/4 Holstein × 3/4 Gir (n = 44), and 1/2 Holstein-Gir (n = 37) crossbred cows, and the pregnancy rate of recipient cows. Viable oocytes were in vitro matured (24 h at 38.8 °C, 5% CO₂ in air) and fertilized by incubating them for 18 to 20 h with frozen-thawed sexed sperm (X-chromosome bearing) from Gir (n = 8) or Holstein (n = 7) sires (2 × 10⁶ sperm/dose). Embryos were cultured in similar conditions of temperature and atmosphere as for IVM, with variable intervals of culture (between Days 2 and 5) completed in a portable incubator. All embryos were transferred fresh, after 24 to 72 h of transportation (up to 2,000 km). On average, 16.7 ± 6.3 oocytes (mean ± SEM) were obtained per OPU procedure and 72.0% were considered viable. Total and viable oocytes per OPU procedure were 17.1 ± 4.5 and 12.1 ± 3.9 for Gir cows, 11.4 ± 3.9 and 8.0 ± 2.7 for Holstein cows, 20.4 ± 5.8 and 16.8 ± 5.0 for 1/4 Holstein × 3/4 Gir, and 31.4 ± 5.6 and 24.3 ± 4.7 for 1/2 Holstein-Gir crossbred females (P < 0.01). The mean number of embryos produced by OPU/IVF and the pregnancy rates were 3.2 (12,243/ 3,778) and 40% for Gir cows, 2.1 (2,426/1,138) and 36% for Holstein cows, 3.9 (1,033/267) and 37% for 1/4 Holstein × 3/4 Gir, and 5.5 (1,222/224), and 37% for 1/2 Holstein-Gir. In conclusion, we compared oocyte yield from two levels of indicus-taurus breeds and demonstrated the efficiency of sexed sperm for in vitro embryo production. Culturing embryos during long distance transportation was successful, with potential for international movement of embryos.     

Functionality of cryopreserved juvenile ovaries from mutant mice in different genetic background strains after allotransplantation

The rapid expansion of mutant mouse colonies for biomedical research has resulted in lack of space at laboratory animal facilities and increasing risks of losing precious lines. These challenges require cheap and effective methods in addition to freezing embryos and sperm to archive the expanding mutant mouse lines. Cryopreservation of mouse ovarian tissue has been reported, but the application in the diverse mutant lines and genetic backgrounds has not yet been studied. In this study, juvenile ovaries (10-day-old) collected from genetically modified mouse lines were cryopreserved using high concentrations of cryoprotectants (dimethyl sulfoxide (Me₂SO) and ethylene glycol (EG)) and instrumented ultra-rapid freezing. The validation of the frozen ovary batches was assessed by orthotopically transplanting a thawed ovary into a nearly completely ovariectomized mature female (congenic with the ovary donor). After 2 weeks of recovery, the ovary recipient was continuously paired with a male (congenic with the ovary donor) to evaluate the fertility of the recipient and delivered offspring were genotyped to evaluate the continued functionality of the grafted ovary. The recipient females delivered genetically modified offspring starting 6 weeks after ovary transplantation and lasting up to 6 months. The presented cryopreservation and transplantation protocols enabled retrieval of the genetic modification in 20 (from 22) genetically modified mutant mouse models on a C57BL/6 (17), FVB (2), or BALB/c (1) background. The thawed ovaries functioned after successful orthotopic allotransplantation to congenic wild-type recipients and produced mutant offspring, which allowed recreation of the desired genotype as a heterozygote on the proper genetic background. The results indicate that cryopreservation of mouse ovaries is a promising method to preserve genetic modification of the increasing number of mutant mouse models and can be used as a model for ovary cryopreservation using a variety of mouse mutants.     

Subtle topographical differences along a floodplain promote different plant strategies among Paspalum dilatatum subspecies and populations

It was hypothesised that subtle topographical differences might cause the existence of ecotypes along a floodplain. The apomict grass Paspalum dilatatum subspecies dilatatum inhabits flood‐prone lowlands as well as nearby uplands in the floodplains of Argentina, while the sexual P. dilatatum subspecies flavescens almost exclusively inhabits the uplands. The aim of the present study was to identify the different traits that allow these P. dilatatum populations to inhabit different habitats. Plants of P. dilatatum were reciprocally transplanted between uplands and lowlands. Morphophysiological traits related to flooding tolerance were measured during a flood. Subspecies dilatatum from the uplands and subspecies flavescens showed a high physiological performance in the uplands but a considerable decrease in stomatal conductance, net photosynthesis rates and tiller number in the flooded lowlands. In contrast, the subspecies dilatatum from the lowlands showed relatively lower and stable stomatal conductance, photosynthesis rates and leaf water potential at both sites. Subspecies dilatatum from the lowlands outperformed upland populations at the lowland site with respect to tillering. Leaves of subspecies dilatatum from the lowlands that had grown at the lowland habitat had a lower blade/sheath proportion than leaves of plants transplanted to the uplands. This behavior did not occur in both upland populations. Results suggest that dilatatum Lowland plants have the typical strategy of stress‐tolerant genotypes and that the upland populations are adapted to habitats where competitive species are selected. In conclusion, habitats with subtle differences in topographic level can favour both ecotypic differentiations within an apomict subspecies but also the maintenance of morphophysiological similitudes between coexisting upland populations belonging to different subspecies.     

The Effect of Restoration Methods on the Quality of the Restoration and Resistance to Invasion by Exotics

The methods employed in a restoration can impact the resulting plant community. This study investigated the effect of restoration method on several indices of plant community structure by comparing two restoration methods conducted over an 8‐year period to a naturally colonized postagricultural field and a remnant grassland. The restoration methods included (1) distributing seed over fallow fields and (2) planting established seedlings in combination with seeding a fallow field. We found greater plant community resemblance (i.e., floristic quality, native species richness, and native diversity) to remnant grasslands with the introduction of seedlings during the first 4 years of restoration. There was also a negative correlation between the native plant diversity and the density of exotic plants in the restoration. This relationship suggests that introducing native plants in postagricultural fields may represent an effective management strategy to reduce exotic plant density.     

Modulation by ellagic acid of DCA-induced developmental toxicity in the zebrafish (Danio rerio)

The ability of ellagic acid (EA) to modulate dichloroacetic acid (DCA)-induced developmental toxicity and oxidative damage was examined in zebrafish embryos. Embryos were exposed to 20 mM EA administered concomitantly with 32 mM DCA at 4 hours postfertilization (hpf) and 20 h later. Embryos were observed through 144 hpf for developmental malformations, and production of superoxide anion (SA) and nitric oxide (NO) was determined in embryonic homogenates. DCA was shown to produce developmental abnormalities and significant levels of SA and NO in zebrafish embryos. EA exposure alleviated the developmental malformations observed in treated embryos and decreased the levels of SA and NO in those same embryos. Less than 10% of DCA + EA exposed embryos showed developmental malformations compared to 100% of embryos treated with DCA alone. Animals in this group that developed malformations were shown to have fewer defects than those treated with DCA only. Taken together, the results confirm the involvement of oxidative stress in the developmental toxicity of DCA in zebrafish embryos, and suggest possible protection against those effects with the use of antioxidants.