Mice carrying a conditional Serca2 allele for the generation of Ca handling-deficient mouse models

Sarco(endo)plasmic reticulum calcium ATPases (SERCA) are cellular pumps that transport Ca²⁺ into the sarcoplasmic reticulum (SR). Serca2 is the most widely expressed gene family member. The very early embryonic lethality of Serca2^null mouse embryos has precluded further evaluation of loss of Serca2 function in the context of organ physiology. We have generated mice carrying a conditional Serca2^flox allele which allows disruption of the Serca2 gene in an organ-specific and/or inducible manner. The model was tested by mating Serca2^flox mice with MLC-2v^wt/Cre mice and with αMHC-Cre transgenic mice. In heterozygous Serca2^wt/floxMLC-2v^wt/Cre mice, the expression of SERCA2a and SERCA2b proteins were reduced in the heart and slow skeletal muscle, in accordance with the expression pattern of the MLC-2v gene. In Serca2^flox/flox Tg(αMHC-Cre) embryos with early homozygous cardiac Serca2 disruption, normal embryonic development and yolk sac circulation was maintained up to at least embryonic stage E10.5. The Serca2^flox mouse is the first murine conditional gene disruption model for the SERCA family of Ca²⁺ ATPases, and should be a powerful tool for investigating specific physiological roles of SERCA2 function in a range of tissues and organs in vivo both in adult and embryonic stages.     

Aspects of cAMP Signaling in Epileptogenesis and Seizures and Its Potential as Drug Target

Neurochemical Research - Epilepsy is one of the most common chronic neurological conditions. Today, close to 30 different medications to prevent epileptic seizures are in use; yet, far from all...     

Grouping horses according to gender—Effects on aggression, spacing and injuries

Many horse owners tend to group horses according to gender, in an attempt to reduce aggressive interactions and the risk of injuries. The aim of our experiment was to test the effects of such gender separation on injuries, social interactions and individual distance in domestic horses. A total of 66 horses were recruited from 4 different farms in Norway and Denmark and divided into six batches. Within each batch, horses were allotted into one mare group, one gelding group and one mixed gender group, with most groups consisting of three or four animals. After 4–6 weeks of acclimatisation, a trained observer recorded all social interactions using direct, continuous observation 1 h in the morning and 1 h in the afternoon for three consecutive days. Recordings of the nearest neighbour of each horse were performed using instantaneous sampling every 10 min. The horses were inspected for injuries before grouping, day 1 after grouping and after 4–6 weeks. No significant effect of gender composition was found on social interactions (P > 0.05), spacing (P > 0.07) or injuries (P > 0.23). Eighty percent of all aggressive interactions recorded were threats, not involving physical contact. Horses with the smallest space allowance showed the highest mean number of aggressive interactions (28.6 ± 6.1 interactions per 6 h) compared to the mean of all the other batches (8.3 ± 1.0 interactions per 6 h). Very few injuries were found and most were superficial. In conclusion, gender composition does not seem to have any effect on aggression level, spacing or injuries. However, the early social experience of horses, management of feeding and space allowance probably represents more important factors for successful group housing of domestic horses.     

Low Omega-3 Index in Pregnancy Is a Possible Biological Risk Factor for Postpartum Depression

Background

Depression is a common disorder affecting 10–15% women in the postpartum period. Postpartum depression can disrupt early mother-infant interaction, and constitutes a risk factor for early child development. Recently, attention has been drawn to the hypothesis that a low intake of seafood in pregnancy can be a risk factor for postpartum depression. Seafood is a unique dietary source of the marine omega-3 fatty acids and is a natural part of a healthy balanced diet that is especially important during pregnancy.

Methods

In a community based prospective cohort in a municipality in Western Norway, we investigated both nutritional and psychological risk factors for postpartum depression. The source population was all women who were pregnant within the period November 2009 - June 2011. The fatty acid status in red blood cells was assessed in the 28^th gestation week and participants were screened for postpartum depression using the Edinburgh Postnatal Depression Scale (EPDS) three months after delivery. The aim of the present study was to investigate if a low omega-3 index in pregnancy is a possible risk factor for postpartum depression.

Results

In a simple regression model, the omega-3 index was associated with the EPDS score in a nonlinear inverse manner with an R square of 19. Thus, the low omega-3 index explained 19% of the variance in the EPDS score. The DPA content, DHA content, omega-3 index, omega-3/omega-6 ratio, total HUFA score, and the omega-3 HUFA score were all inversely correlated with the EPDS score. The EPDS scores of participants in the lowest omega-3 index quartile were significantly different to the three other omega-3 index quartiles.

Conclusion

In this study population, a low omega-3 index in late pregnancy was associated with higher depression score three months postpartum.     

Characteristics of Glucose Metabolism in Nordic and South Asian Subjects with Type 2 Diabetes

Background

Insulin resistance and type 2 diabetes are more prevalent in people of South Asian ethnicity than in people of Western European origin. To investigate the source of these differences, we compared insulin sensitivity, insulin secretion, glucose and lipid metabolism in South Asian and Nordic subjects with type 2 diabetes.

Methods

Forty-three Nordic and 19 South Asian subjects with type 2 diabetes were examined with intra-venous glucose tolerance test, euglycemic clamp including measurement of endogenous glucose production, indirect calorimetry measuring glucose and lipid oxidation, and dual x-ray absorptiometry measuring body composition.

Results

Despite younger mean ± SD age (49.7±9.4 vs 58.3±8.3 years, p = 0.001), subjects of South Asian ethnicity had the same diabetes duration (9.3±5.5 vs 9.6±7.0 years, p = 0.86), significantly higher median [inter-quartile range] HbA_1c (8.5 [1.6] vs 7.3 [1.6] %, p = 0.024) and lower BMI (28.7±4.0 vs 33.2±4.7 kg/m², p<0.001). The South Asian group exhibited significantly higher basal endogenous glucose production (19.1 [9.1] vs 14.4 [6.8] µmol/kgFFM⋅min, p = 0.003). There were no significant differences between the groups in total glucose disposal (39.1±20.4 vs 39.2±17.6 µmol/kgFFM⋅min, p = 0.99) or first phase insulin secretion (AUC_{0–8 min}: 220 [302] vs 124 [275] pM, p = 0.35). In South Asian subjects there was a tendency towards positive correlations between endogenous glucose production and resting and clamp energy expenditure.

Conclusions

Subjects of South Asian ethnicity with type 2 diabetes, despite being younger and leaner, had higher basal endogenous glucose production, indicating higher hepatic insulin resistance, and a trend towards higher use of carbohydrates as fasting energy substrate compared to Nordic subjects. These findings may contribute to the understanding of the observed differences in prevalence of type 2 diabetes between the ethnic groups.     

The sub-ice algal community in the Chukchi sea: large- and small-scale patterns of abundance based on images from a remotely operated vehicle

We examined the sub-ice algal community in the Chukchi Sea during June 1998 using a remotely operated vehicle (ROV). Ice algae were observed on the under-ice surface at all ten stations (from 70°29′N to 72°26′N; 162°00′W to 153°56′W) and varied in abundance and distribution from small aggregations limited to depressions in the ice to nets, curtains and strands of Melosira. There was no relationship between percent cover of sub-ice algae and physical factors at the kilometer scale, but at the scale of individual ice floes the percent cover of sub-ice algae was positively correlated with distance from the floe edge and negatively correlated with snow depth. A significant positive relationship between the concentration of sediment pigments and percent cover of sub-ice could indicate a coupling between ice algal and benthic systems. Pieces of ice algae that appeared to be Melosira were observed on the seafloor to a depth of over 100 m and cells or spores of obligate ice algal taxa were collected from sediments from 44-m to 1,000-m deep. The large biomass of sub-ice algae observed at many stations in the Chukchi Sea and the presence of ice algae on the seafloor indicates that the distribution and abundance of sub-ice algae needs to be understood if we are to evaluate the role of ice algae in the Arctic marine ecosystem.     

Structural Basis for dsRNA Recognition,Filament Formation,and Antiviral Signal Activation by MDA5

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Highlights? MDA5 forms an open, C-shaped ring around the viral dsRNA stem ? The CTD of MDA5 has a different orientation and flexibility compared to RIG-I ? MDA5 forms filaments by stacking monomers head-to-tail with a 70° turn per monomer ? The 2CARD domain assembles into oligomers that activate interferon signaling via MAVS     

Therapeutic vaccination against autologous cancer stem cells with mRNA-transfected dendritic cells in patients with glioblastoma

Background

The growth and recurrence of several cancers appear to be driven by a population of cancer stem cells (CSCs). Glioblastoma, the most common primary brain tumor, is invariably fatal, with a median survival of approximately 1 year. Although experimental data have suggested the importance of CSCs, few data exist regarding the potential relevance and importance of these cells in a clinical setting.

Methods

We here present the first seven patients treated with a dendritic cell (DC)-based vaccine targeting CSCs in a solid tumor. Brain tumor biopsies were dissociated into single-cell suspensions, and autologous CSCs were expanded in vitro as tumorspheres. From these, CSC-mRNA was amplified and transfected into monocyte-derived autologous DCs. The DCs were aliquoted to 9–18 vaccines containing 10⁷ cells each. These vaccines were injected intradermally at specified intervals after the patients had received a standard 6-week course of post-operative radio-chemotherapy. The study was registered with the ClinicalTrials.gov identifier NCT00846456.

Results

Autologous CSC cultures were established from ten out of eleven tumors. High-quality RNA was isolated, and mRNA was amplified in all cases. Seven patients were able to be weaned from corticosteroids to receive DC immunotherapy. An immune response induced by vaccination was identified in all seven patients. No patients developed adverse autoimmune events or other side effects. Compared to matched controls, progression-free survival was 2.9 times longer in vaccinated patients (median 694 vs. 236 days, p = 0.0018, log-rank test).

Conclusion

These findings suggest that vaccination against glioblastoma stem cells is safe, well-tolerated, and may prolong progression-free survival.     

Growth rate,health and welfare in a dairy herd with natural suckling until 6–8 weeks of age: a case report

Over a period of two years, growth rate and health were measured for dairy calves allowed to suckle their mothers up to 6–8 weeks of age. Thirty-one calves were weighted weekly, and the mean daily growth rate was 1.2 ± 0.03 kg from birth up to 13 weeks of age. Illness in calves and young stock was not observed. In the cows, the mean incidences of ketosis, displaced abomasum, puerperal paresis, mastitis, teat injury and retained placenta were 0, 0, 8, 22, 1 and 1%, respectively, during a 6-year period. The mean daily gain of 56 growing bulls was 1.4 kg when slaughtered at 15 months of age, which is higher than the mean daily gain of 0.95 kg in the population. Probiotics, hormones and vaccines were not used, and antibiotics were only used for treating illness. The present study indicates many advantages and few problems when dairy calves are penned together with the cows and allowed natural feeding up to 6–8 weeks of age. This production system was easy to manage, preferred by the farmer, and may satisfy the public concern regarding the practice of immediate separation of cow and calf in commercial milk production.