A comparison of anther and microspore culture as a breeding tool in Brassica napus

Summary A direct comparison of microspore culture and anther culture was made in Brassica napus using F₁ crosses of Regent (canola) by Golden (rapeseed), and their reciprocals, as well as a hybrid between Reston and a highly embryogenic, canola-quality breeding line (G231) as donor plants. The study confirmed that microspore culture can be ten times more efficient than anther culture for embryo production. Embryo yields from cultures initiated from the Reston x G231 were four-fold greater than those initiated from the Regent x Golden crosses, and significant differences were also detected among cultures initiated from the different Regent x Golden crosses. These results illustrate the influence that donor plant genotype has on embryo production. However, superior embryogenic potential among donor material was not always coincident with superior plant production. The average haploid-todiploid ratio in microspore-derived regenerates was 21 for the population obtained from the Regent x Golden crosses but 11 for the Reston x G231 cross. For both types of material, the frequency of diploids increased upon repeated cycles of explanting. A field study showed that there were no differences between the populations of anther-derived and microspore-derived spontaneous diploid and doubled haploid lines, with respect to the days required for them to flower or to mature. The information is valuable for canola breeding programs considering the use of haploidy.     

Wabusk of the Omushkegouk: Cree-Polar Bear (Ursus maritimus) Interactions in Northern Ontario

In order to understand wabusk (polar bear, Ursus maritimus) behaviours and interactions with people in the Hudson Bay lowlands of northern Ontario we conducted this collaborative study of Cree kiskayndamowin/knowledge. Our findings reveal that Cree knowledge supports previously published information on polar bears, while adding further contextual findings: that male polar bears travel greater distances into the muskeg than previously recorded; that wabusk prey on amisk (beaver, Castor canadensis); that wabusk interact with muskwa (black bears, Ursus americanus); and that human-polar bear interactions occur in this region of northern Canada. Bearing in mind that Cree knowledge has been recognized in wildlife management strategies (i.e., for beaver, caribou and moose) elsewhere in Canada, this particular body of information is timely, especially since polar bears are considered threatened under the Ontario Endangered Species Act, and the province is developing a recovery strategy for the Southern Hudson Bay polar bear population. The federal government is also contemplating listing polar bears in Canada as a “species of special concern” under the Species at Risk Act (SARA). While it is unclear as to how these decisions will impact the Cree-polar interactions, the listing of polar bears by both governments, but especially the provincial government of Ontario, must recognize treaty and Aboriginal rights, acknowledge its duties to consult and properly accommodate Aboriginal people’s views, incorporate Cree kiskayndamowin/knowledge of wabusk, and re-examine the proposed Wabusk Co-Management Agreement draft developed by the Ontario Ministry of Natural Resources (OMNR) and Coastal Cree First Nations of Northern Ontario in 1994. The article provides recommendations that highlight how the Northern Cree First Nations, through the development of the Recovery Strategy for Polar Bear in Ontario, can become engaged in the management of wabusk in Ontario and throughout Canada.     

Up-regulation of mesotocin receptors in the tammar wallaby myometrium is pregnancy-specific and independent of estrogen

The oxytocin-like peptide of most Australian marsupials is mesotocin, which stimulates uterine contractions and is important for normal birth in the tammar wallaby. Female marsupials have two uteri and, in monovular species such as the tammar, one uterus is gravid with a single fetus, whereas the contralateral uterus is nongravid. A significant increase in myometrial mesotocin receptor concentrations occurs only in the gravid uterus on Day 23 of the 26-day gestation. This study examined whether or not mesotocin receptors are present in the myometrium and are up-regulated at the equivalent stage of the luteal phase in unmated tammars. In contrast to the marked increase in mesotocin receptor mRNA and protein concentrations in the myometrium of the gravid uterus during pregnancy, receptors did not increase in the unmated animals. There were also no significant differences between the two uteri, except on Day 27. Plasma profiles of peripheral estradiol-17beta and progesterone did not differ significantly between pregnant and nonpregnant cycles. However, progesterone concentrations were significantly lower on Day 1 postpartum compared with Day 27 of the nonpregnant cycle. In pregnant tammars, the molar ratio of circulating estradiol-17beta to progesterone increased significantly between Day 25 of gestation and 1 day postpartum, but was not correlated with an increase in mesotocin receptor concentrations in either uterus. The data confirm that a local fetal influence is more important than systemic factors, such as estrogen, in the regulation of uterine mesotocin receptors in the tammar wallaby.     

An antibody to Notch3 reverses the skeletal phenotype of lateral meningocele syndrome in male mice

Lateral meningocele syndrome (LMS), a genetic disorder characterized by meningoceles and skeletal abnormalities, is associated with NOTCH3 mutations. We created a mouse model of LMS (Notch3^tm1.1Ecan) by introducing a tandem termination codon in the Notch3 locus upstream of the proline (P), glutamic acid (E), serine (S) and threonine (T) domain. Microcomputed tomography demonstrated that Notch3^tm1.1Ecan mice exhibit osteopenia. The cancellous bone osteopenia was no longer observed after the intraperitoneal administration of antibodies directed to the negative regulatory region (NRR) of Notch3. The anti-Notch3 NRR antibody suppressed the expression of Hes1, Hey1, and Hey2 (Notch target genes), and decreased Tnfsf11 (receptor activator of NF Kappa B ligand) messenger RNA in Notch3^tm1.1Ecan osteoblast (OB) cultures. Bone marrow-derived macrophages (BMMs) from Notch3^tm1.1Ecan mutants exhibited enhanced osteoclastogenesis in culture, and this was increased in cocultures with Notch3^tm1.1Ecan OB. Osteoclastogenesis was suppressed by anti-Notch3 NRR antibodies in Notch3^tm1.1Ecan OB/BMM cocultures. In conclusion, the cancellous bone osteopenia of Notch3^tm1.1Ecan mutants is reversed by anti-Notch3 NRR antibodies.     

Skeletal Muscle Insulin Resistance Associated with Cholesterol-Induced Activation of Macrophages Is Prevented by High Density Lipoprotein

Background

Emerging evidence suggests that high density lipoprotein (HDL) may modulate glucose metabolism through multiple mechanisms including pancreatic insulin secretion as well as insulin-independent glucose uptake into muscle. We hypothesized that HDL may also increase skeletal muscle insulin sensitivity via cholesterol removal and anti-inflammatory actions in macrophages associated with excess adiposity and ectopic lipid deposition.

Methods

Human primary and THP-1 macrophages were treated with vehicle (PBS) or acetylated low density lipoprotein (acLDL) with or without HDL for 18 hours. Treatments were then removed, and macrophages were incubated with fresh media for 4 hours. This conditioned media was then applied to primary human skeletal myotubes derived from vastus lateralis biopsies taken from patients with type 2 diabetes to examine insulin-stimulated glucose uptake.

Results

Conditioned media from acLDL-treated primary and THP-1 macrophages reduced insulin-stimulated glucose uptake in primary human skeletal myotubes compared with vehicle (primary macrophages, 168±21% of basal uptake to 104±19%; THP-1 macrophages, 142±8% of basal uptake to 108±6%; P<0.05). This was restored by co-treatment of macrophages with HDL. While acLDL increased total intracellular cholesterol content, phosphorylation of c-jun N-terminal kinase and secretion of pro- and anti-inflammatory cytokines from macrophages, none were altered by co-incubation with HDL. Insulin-stimulated Akt phosphorylation in human skeletal myotubes exposed to conditioned media was unaltered by either treatment condition.

Conclusion

Inhibition of insulin-stimulated glucose uptake in primary human skeletal myotubes by conditioned media from macrophages pre-incubated with acLDL was restored by co-treatment with HDL. However, these actions were not linked to modulation of common pro- or anti-inflammatory mediators or insulin signaling via Akt.     

Physiological state co-regulates thousands of mammalian mRNA splicing events at tandem splice sites and alternative exons

Thousands of tandem alternative splice sites (TASS) give rise to mRNA insertion/deletion variants with small size differences. Recent work has concentrated on the question of biological relevance in general, and the physiological regulation of TASS in particular. We have quantitatively studied 11 representative TASS cases in comparison to one mutually exclusive exon case and two cassette exons (CEs) using a panel of human and mouse tissues, as well as cultured cell lines. Tissues show small but significant differences in TASS isoform ratios, with a variance 4- to 20-fold lower than seen for CEs. Remarkably, in cultured cells, all studied alternative splicing (AS) cases showed a cell-density-dependent shift of isoform ratios with similar time series profiles. A respective genome-wide co-regulation of TASS splicing was shown by next-generation mRNA sequencing data. Moreover, data from human and mouse organs indicate that this co-regulation of TASS occurs in vivo, with brain showing the strongest difference to other organs. Together, the results indicate a physiological AS regulation mechanism that functions almost independently from the splice site context and sequence.     

The effext of papain, neuraminidase and AET on the behavior of lymphocytes in cytotoxicity tests]

The influence of papain, neuraminidase and AET to lymphocytes is reported. All three substances strengthen the lymphocytotoxic reaction in low concentrations or by short incubation time and lower it in higher concentrations or longer incubation since. Membrane changes by these three substances are reversible under defined conditions.     

“Celtic migrations”: Fact or fiction? Strontium and oxygen isotope analysis of the Czech cemeteries of Radovesice and Kutná Hora in Bohemia

Strontium and oxygen isotope analysis of human remains from the early La Tène (fourth/third century BC) Czech cemeteries of Radovesice I (RAD I), Radovesice II (RAD II), and Kutná Hora were conducted to investigate the importance of residential changes during the period of the historic “Celtic migrations”. In the initial phases (LT A/B), the grave goods of these cemeteries are typical for the core area of the La Tène culture, while around 300 BC (LT B2) an alteration occurs and typical Bohemian styles arise, and connections to Moravia and the Danubian region become visible. The strontium isotope ratios are highly varied with ⁸⁷Sr/⁸⁶Sr values between 0.7062 and 0.7153 in Radovesice, and between 0.7082 and 0.7147 in Kutná Hora. The oxygen isotope data are more homogeneous and yield δ¹⁸O_p ratios from 14.8‰ to 17.2‰ [mean: 16.2‰ ± 0.5 (1σ)] in Radovesice, and from 14.9‰ to 17.3‰ [mean: 16.5‰ ± 0.6 (1σ)] in Kutná Hora. Because the geological properties of the landscapes around the sites are variable and complex, most of the observed variations among the strontium isotope ratios may have been caused by agricultural practices, such as regularly changing farming land. Nevertheless, there are some individuals who differ completely from the regional isotopic baseline values. This suggests that at least a small part of the community migrated, which does not seem to be correlated with any particular phase of the La Tène period. Remarkably, it is mainly males who seem to be of nonlocal origin, and particularly those who were buried as warriors. Females, on the other hand, appear to have been more closely bonded to the Bohemian region. Whether the “foreign” individuals with differing isotopic compositions came from Moravia or the Danubian region remains debatable. Am J Phys Anthropol 155:496–512, 2014. © 2014 Wiley Periodicals, Inc.     

Defining hotspots of characteristic species for multiple taxonomic groups in the Netherlands

Biogeographical zonation based on single taxa poses major limitations on planning for nature conservation. This paper identifies biogeographical patterns of multiple taxa in the Netherlands, where no endemics are present at species level, on the basis of characteristic species. We used occurrence data on five species groups in order to identify spatially coherent, ecologically important regions. TWINSPAN was used to cluster grid squares according to similarity in species composition for each taxonomic group. Species that are characteristic of each of the clusters were identified using a preference index, and corresponding clusters among the taxonomic groups were identified with Kappa statistics. Regions containing characteristic species for several taxonomic groups were defined as ‘hotspots’. Stepwise discriminant analysis was then used to characterize these hotspots according to differences in environmental conditions. The analysis yielded five regions that are clearly distinct in terms of species composition for individual taxonomic groups. Each region is characterized by a set of unique species that occur in the zonation of at least two of the taxonomic groups. Stepwise discriminant analysis revealed significant environmental differences among these regions. The concept of hotspots as operationalized in this study can make nature conservation planning more efficient. In combination, the hotspots defined here comprise the majority of the species occurring in the Netherlands for the studied groups. Therefore, this regionalization should be taken into account when prioritizing nature conservation efforts.