Notch2 governs the rate of generation of mouse long- and short-term repopulating stem cells

HSCs either self-renew or differentiate to give rise to multipotent cells whose progeny provide blood cell precursors. However, surprisingly little is known about the factors that regulate this choice of self-renewal versus differentiation. One candidate is the Notch signaling pathway, with ex vivo studies suggesting that Notch regulates HSC differentiation, although a functional role for Notch in HSC self-renewal in vivo remains controversial. Here, we have shown that Notch2, and not Notch1, inhibits myeloid differentiation and enhances generation of primitive Sca-1(+)c-kit(+) progenitors following in vitro culture of enriched HSCs with purified Notch ligands. In mice, Notch2 enhanced the rate of formation of short-term repopulating multipotential progenitor cells (MPPs) as well as long-term repopulating HSCs, while delaying myeloid differentiation in BM following injury. However, consistent with previous reports, once homeostasis was achieved, neither Notch1 nor Notch2 affected repopulating cell self-renewal. These data indicate a Notch2-dependent role in assuring orderly repopulation by HSCs, MPPs, myeloid cells, and lymphoid cells during BM regeneration.     

Variable velocity encoding in a three‐dimensional,three‐directional phase contrast sequence: Evaluation in phantom and volunteers

Purpose:

To evaluate accuracy and noise properties of a novel time‐resolved, three‐dimensional, three‐directional phase contrast sequence with variable velocity encoding (denoted 4D‐vPC) on a 3 Tesla MR system, and to investigate potential benefits and limitations of variable velocity encoding with respect to depicting blood flow patterns.

Materials and Methods:

A 4D PC‐MRI sequence was modified to allow variable velocity encoding (VENC) over the cardiac cycle in all three velocity directions independently. 4D‐PC sequences with constant and variable VENC were compared in a rotating phantom with respect to measured velocities and noise levels. Additionally, comparison of flow patterns in the ascending aorta was performed in six healthy volunteers.

Results:

Phantom measurements showed a linear relationship between velocity noise and velocity encoding. 4D‐vPC MRI presented lower noise levels than 4D‐PC both in phantom and in volunteer measurements, in agreement with theory. Volunteer comparisons revealed more consistent and detailed flow patterns in early diastole for the variable VENC sequences.

Conclusion:

Variable velocity encoding offers reduced noise levels compared with sequences with constant velocity encoding by optimizing the velocity‐to‐noise ratio (VNR) to the hemodynamic properties of the imaged area. Increased VNR ratios could be beneficial for blood flow visualizations of pathology in the cardiac cycle. J. Magn. Reson. Imaging 2012; 36:1450–1459. © 2012 Wiley Periodicals, Inc.     

Free-breathing imaging of the heart using 2D cine-GRICS (generalized reconstruction by inversion of coupled systems) with assessment of ventricular volumes and function

Purpose:

To assess cardiac function by means of a novel free‐breathing cardiac magnetic resonance imaging (MRI) strategy.

Materials and Methods:

A stack of ungated 2D steady‐state free precession (SSFP) slices was acquired during free breathing and reconstructed as cardiac cine imaging based on the generalized reconstruction by inversion of coupled systems (GRICS). A motion‐compensated sliding window approach allows reconstructing cine movies with most motion artifacts cancelled. The proposed reconstruction uses prior knowledge from respiratory belts and electrocardiogram recordings and features a piecewise linear model that relates the electrocardiogram signal to cardiac displacements. The free‐breathing protocol was validated in six subjects against a standard breath‐held protocol.

Results:

Image sharpness, as assessed by the image gradient entropy, was comparable to that of breath‐held images and significantly better than in uncorrected images. Volumetric parameters of cardiac function in the left ventricle (LV) and right ventricle (RV) were similar, including end‐systolic volumes, end‐diastolic volumes and mass, stroke volumes, and ejection fractions (with differences of 3% ± 2.4 in the LV and 2.9% ± 4.4 in the RV). The duration of the free‐breathing protocol was nearly the same as the breath‐held protocol.

Conclusion:

Free‐breathing cine‐GRICS enables accurate assessment of volumetric parameters of cardiac function with efficient correction of motion. J. Magn. Reson. Imaging 2012;340‐351. © 2011 Wiley Periodicals, Inc.     

Role of angiotensin II AT1 receptor blockers in the treatment of arterial hypertension

Hypertension is a very common condition and the most important risk factor for the occurrence of cardiovascular events. The hyperactivity of the renin-angiotensin-aldosterone system is considered a cardiovascular risk factor in subjects with essential hypertension. The intrinsic vascular abnormality in which the renin-angiotensin-aldosterone system is clearly the milieu for the development of the pathologic changes in blood vessel walls is one of the causes of the establishment of hypertension. Many drugs with different mechanisms of action have been used for the treatment of hypertension and its vascular complications. Nevertheless, the utilities of many drugs are limited by their adverse effects. Continuous research in the search for new pharmacological agents for the treatment of hypertension has led to the development of angiotensin II receptor type AT1 blockers. The most important functions mediated by AT1 receptors include: vasoconstriction, induction of the production and release of aldosterone, renal reabsorption of sodium, cardiac cellular growth, proliferation of vascular smooth muscle, increase of peripheral noradrenergic action and the central activity of the sympathetic nervous system, stimulation of vasopressin release, and inhibition of renin release from the kidney. The angiotensin II receptor type AT1 blockers inhibit the interaction of angiotensin II with its AT1 receptor. These agents lower blood pressure without producing cough as a side effect since, unlike the angiotensin-converting enzyme inhibitors they do not influence the levels of bradykinin or substance P. Hence, these drugs are suitable for the treatment of hypertensive patients who require therapy with a drug blocking the effect of angiotensin-converting enzyme but cannot use angiotensin-converting enzyme inhibitors due to cough as a side effect.     

Use of bone biochemical markers with dual-energy x-ray absorptiometry for early determination of bone loss in persons with spinal cord injury

Our cross-sectional study aimed at the early determination of changes in bone metabolism in terms of bone mineral density (BMD) and bone turnover in persons with complete spinal cord injury (SCI) during the acute phase of paraplegia. Combined dual-energy x-ray absorptiometry (DXA) and specific biochemical markers of bone turnover were used to determine bone metabolism. Seven persons with SCI (age, 31.3 +/- 9.5 years) who had sustained injury an average of 3 months earlier (103 +/- 10.8 days) were compared with 10 able-bodied controls (27.5 +/- 4.3 years). Four paraplegics and 3 quadriplegics composed the SCI group. BMD was measured by DXA, while bone turnover was evaluated by serum osteocalcin (OC), bone alkaline phosphatase (B-ALP), and serum and urinary type I collagen C-telopeptide (CTXs and CTXu). Regional BMD (proximal femur, lumbar spine, radius, lower limb) was similar in the 2 groups except in the upper limb (P <.05). CTXs and CTXu were significantly higher in SCI (P <.01 and P <.001, respectively), whereas among the bone formation markers used, only serum OC was affected by immobilization (P <.05). The SCI group developed hypercalciuria (0.76 +/- 0.37 v 0.35 +/- 0.14), whereas calcemia was normal (2.42 +/- 0.09 v 2.31 +/- 0.10). Intact parathyroid hormone (iPTH) and 1.25 (OH)(2) vitamin D levels were suppressed in persons with SCI (P <.001) by 80.6% and 66%, respectively. In conclusion, it was not possible to detect any variation in BMD from the DXA technique at this early stage of demineralization, but the sensitivity and early response of the biochemical markers strongly suggested their usefulness for the early identification of persons with SCI at risk of severe osteoporosis.     

The Levels of Brachyspira hyodysenteriae Binding to Porcine Colonic Mucins Differ between Individuals,and Binding Is Increased to Mucins from Infected Pigs with De Novo MUC5AC Synthesis

Brachyspira hyodysenteriae colonizes the pig colon, resulting in mucohemorrhagic diarrhea and growth retardation. Fecal mucus is a characteristic feature of swine dysentery; therefore, we investigated how the mucin environment changes in the colon during infection with B. hyodysenteriae and how these changes affect this bacterium''s interaction with mucins. We isolated and characterized mucins, the main component of mucus, from the colon of experimentally inoculated and control pigs and investigated B. hyodysenteriae binding to these mucins. Fluorescence microscopy revealed a massive mucus induction and disorganized mucus structure in the colon of pigs with swine dysentery. Quantitative PCR (qPCR) and antibody detection demonstrated that the mucus composition of pigs with swine dysentery was characterized by de novo expression of MUC5AC and increased expression of MUC2 in the colon. Mucins from the colon of inoculated and control pigs were isolated by two steps of isopycnic density gradient centrifugation. The mucin densities of control and inoculated pigs were similar, whereas the mucin quantity was 5-fold higher during infection. The level of B. hyodysenteriae binding to mucins differed between pigs, and there was increased binding to soluble mucins isolated from pigs with swine dysentery. The ability of B. hyodysenteriae to bind, measured in relation to the total mucin contents of mucus in sick versus healthy pigs, increased 7-fold during infection. Together, the results indicate that B. hyodysenteriae binds to carbohydrate structures on the mucins as these differ between individuals. Furthermore, B. hyodysenteriae infection induces changes to the mucus niche which substantially increase the amount of B. hyodysenteriae binding sites in the mucus.     

Veal Calves Produce Less Antibodies against C. Perfringens Alpha Toxin Compared to Beef Calves

Enterotoxaemia is a disease with a high associated mortality rate, affecting beef and veal calves worldwide, caused by C. perfringens alpha toxin and perfringolysin. A longitudinal study was conducted to determine the dynamics of antibodies against these toxins in 528 calves on 4 beef and 15 veal farms. The second study aimed to determine the effect of solid feed intake on the production of antibodies against alpha toxin and perfringolysin. The control group only received milk replacer, whereas in the test group solid feed was provided. Maternal antibodies for alpha toxin were present in 45% of the veal calves and 66% of the beef calves. In beef calves a fluent transition from maternal to active immunity was observed for alpha toxin, whereas almost no veal calves developed active immunity. Perfringolysin antibodies significantly declined both in veal and beef calves. In the second study all calves were seropositive for alpha toxin throughout the experiment and solid feed intake did not alter the dynamics of alpha and perfringolysin antibodies. In conclusion, the present study showed that veal calves on a traditional milk replacer diet had significantly lower alpha toxin antibodies compared to beef calves in the risk period for enterotoxaemia, whereas no differences were noticed for perfringolysin.