Comparison of diffusion MRI and CLARITY fiber orientation estimates in both gray and white matter regions of human and primate brain.

Diffusion MRI (dMRI) represents one of the few methods for mapping brain fiber orientations non-invasively. Unfortunately, dMRI fiber mapping is an indirect method that relies on inference from measured diffusion patterns. Comparing dMRI results with other modalities is a way to improve the interpretation of dMRI data and help advance dMRI technologies. Here, we present methods for comparing dMRI fiber orientation estimates with optical imaging of fluorescently labeled neurofilaments and vasculature in 3D human and primate brain tissue cuboids cleared using CLARITY. The recent advancements in tissue clearing provide a new opportunity to histologically map fibers projecting in 3D, which represents a captivating complement to dMRI measurements. In this work, we demonstrate the capability to directly compare dMRI and CLARITY in the same human brain tissue and assess multiple approaches for extracting fiber orientation estimates from CLARITY data. We estimate the three-dimensional neuronal fiber and vasculature orientations from neurofilament and vasculature stained CLARITY images by calculating the tertiary eigenvector of structure tensors. We then extend CLARITY orientation estimates to an orientation distribution function (ODF) formalism by summing multiple sub-voxel structure tensor orientation estimates. In a sample containing part of the human thalamus, there is a mean angular difference of 19o±15o between the primary eigenvectors of the dMRI tensors and the tertiary eigenvectors from the CLARITY neurofilament stain. We also demonstrate evidence that vascular compartments do not affect the dMRI orientation estimates by showing an apparent lack of correspondence (mean angular difference = 49o±23o) between the orientation of the dMRI tensors and the structure tensors in the vasculature stained CLARITY images. In a macaque brain dataset, we examine how the CLARITY feature extraction depends on the chosen feature extraction parameters. By varying the volume of tissue over which the structure tensor estimates are derived, we show that orientation estimates are noisier with more spurious ODF peaks for sub-voxels below 30 µm3 and that, for our data, the optimal gray matter sub-voxel size is between 62.5 µm3 and 125 µm3. The example experiments presented here represent an important advancement towards robust multi-modal MRI-CLARITY comparisons.

Anisotropic longitudinal electronic relaxation affects DNP at cryogenic temperatures.

We report the observation of anisotropic longitudinal electronic relaxation in nitroxide radicals under typical dynamic nuclear polarization conditions. This anisotropy affects the efficiency of dynamic nuclear polarization at cryogenic temperatures of 4 K and high magnetic fields of 6.7 T. Under our experimental conditions, the electron paramagnetic resonance spectrum of nitroxides such as TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl) is only partly averaged by electronic spectral diffusion, so that the relaxation times T1e(ω) vary across the spectrum. We demonstrate how the anisotropy of T1e(ω) can be taken into account in simple DNP models.

Influence of strain and parity on the risk of litter loss in laboratory mice.

Pup mortality is a considerable problem in laboratory mouse breeding and the view that parity influence survival of newborn mice is widespread. Some evidence suggests that maternal behaviour is related to offspring mortality in mice. Parental experience is a factor that can improve maternal behaviour and offspring survival in some mammals. However, few papers report a relationship between parity and pup survival in mice. We investigated the influence of strain and parity on loss of entire litters of C57BL/6 and BALB/c mice using data from a breeding colony. In total, 344 C57BL/6 and 146 BALB/c litters were included. We found a considerable mortality rate for both strains: 32% of C57BL/6 litters and 20% for BALB/c litters were lost. There was a significant difference in survival of the first litter between strains, with 3.6 times higher odds of mortality in C57BL/6 mice (p = 0.0028). Parity or previous parental experience of litter loss did, however, not affect litter loss. The scientific literature does not provide a clear picture of perinatal mortality in laboratory mice. Very few studies report perinatal mortality, and only a handful of papers exist where mortality was systematically studied; this area is thus poorly understood. If perinatal mortality in mice is not recognized and investigated, but instead considered normal when breeding mice, a serious welfare problem might be overlooked.

Effects of conjugated linoleic acids fed to dairy cows during early gestation on hematological, immunological, and metabolic characteristics of cows and their calves.

The aim of the present experiment was to test the stimulation ability of peripheral blood mononuclear cells (PBMC) expressed as stimulation index (SI) of newborn calves and of their dams fed a control fat supplement (CON, n=6) or 50 and 100g/d of a CLA-containing fat supplement (CLA50, n=5, and CLA100, n=6, respectively) during the preceding lactation period for 182 d after calving. The total intake of cis-9,trans-11 and trans-10,cis-12 CLA by groups CLA50 and CLA100 amounted to 4 and 8 g/d each, respectively. For this purpose, blood was collected immediately after parturition from calves before and after colostrum intake, and from cows after parturition and 21 d later. The SI was related to the fatty acid composition of erythrocyte and milk lipids and to various hematological and clinical-chemical parameters. Retrospective evaluation revealed that depletion time (i.e., the individual period elapsed between the day of terminating the feeding of the experimental diet in the preceding lactation period and the day of calving) ranged from 190 to 262 d, which corresponded to fetal exposure times of 19 to 102 d. The SI from cows increased significantly by 77 and 55%, within 21 d after calving according to the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) and Alamar Blue assays, respectively. However, feeding of 50 g of the CLA product failed to demonstrate this increase in the MTT assay. Moreover, SI was significantly lower for calves whose dams belonged to the CLA50 group, whereas stimulation ability was comparable for the PBMC from calves whose mothers were treated with CON and CLA100. Plasma metabolites (total bilirubin, total cholesterol, glucose, nonesterified fatty acids, 3-ß-hydroxybutyrate, total protein, and albumin) and hematological parameters (hematocrit, white blood cell profile) were not significantly influenced by dietary treatments of the cows in the preceding lactation period. Although the fatty acid pattern of erythrocyte lipids of cows remained uninfluenced, that of calves showed alterations due to the feeding type of their dams. For example, C16:0 increased significantly from 14.4 to 16.9% of total fatty acid methyl esters, whereas cis-9,trans-11 CLA increased slightly from 0.11 to 0.15% at the same time in calves when their mothers were fed the CLA100 instead of the CON diet. Fatty acid profile of colostrum was significantly different from that of milk after 3 wk for most of the detected fatty acids, but was not influenced by diet type. In conclusion, feeding a CLA-containing fat supplement during the preceding lactation and gestation period exerted effects on the stimulation ability of PBMC from cows and calves for the subsequent parturition. However, CLA dose effects were inconsistent and require further investigation.

Effect of trans-10, cis-12 conjugated linoleic acid on performance, adipose depot weights, and liver weight in early-lactation dairy cows.

In feeding practice, conjugated linoleic acid (CLA) supplements are used to decrease milk fat excretion in early-lactation dairy cows to save energy to counteract the physiological negative energy balance. The present study was conducted to examine the effects of CLA on energy metabolism, changes in liver weight, and the weight of different adipose depots during early lactation. Primiparous lactating German Holstein cows (n=25) were divided into 5 groups and each group contained 5 animals. The experiment started 21 d prepartum and continued until 105 d in milk (DIM). Cows were slaughtered at 1, 42, and 105 DIM. The experiment was divided into a prepartum period (21 d prepartum until calving), period 1 (1 until 42 DIM), and period 2 (>42 until 105 DIM). In the prepartum period, all animals were housed together and fed the same diet with no CLA supplementation. At 1 DIM, an initial group, with no CLA supplementation, was slaughtered. The 20 remaining cows were assigned to 2 diets. One group received 100g/d of a control fat supplement (CON; n=10) and the other group 100g/d of a CLA supplement (CLA; n=10) from 1 DIM until slaughter. Five cows of each feeding group were slaughtered after 42 DIM and the remaining animals after 105 DIM. The CLA supplement contained approximately 10% each of trans-10, cis-12 CLA and cis-9, trans-11 CLA. During the slaughter process the empty body weight was recorded and the omental, mesenteric, retroperitoneal, and s.c. adipose depots, as well as the liver, were dissected and weighed. The CLA treatment decreased milk fat content in period 1 (14.1%). In period 2, milk fat content (25.4%) and yield (17.1%) were lower in the CLA group. No effect of CLA on milk yield was observed. The net energy intake, milk energy output, and the calculated energy balance remained unchanged by CLA supplementation. No effect of CLA on the weights of liver, omental, mesenteric, or s.c. adipose depots was observed when related to empty body weight. Liver weight increased with DIM, whereas the retroperitoneal adipose depot weight decreased at the same time. Compared with the initial group, the retroperitoneal adipose depot weight for control animals slaughtered after 42 DIM was decreased (47.7%); however, for the CLA group slaughtered after 42 DIM, a trend to a lower retroperitoneal adipose depot weight (34.0%) was observed. This suggests a CLA-induced deceleration of mobilization of the retroperitoneal adipose depot during the first 42 DIM.

Hormone-sensitive lipase protein expression and extent of phosphorylation in subcutaneous and retroperitoneal adipose tissues in the periparturient dairy cow.

Lipomobilization is essential for dairy cows to balance the energy requirement for milk production in early lactation. This study aimed to determine the role of hormone-sensitive lipase (HSL) and its activation by phosphorylation at Ser 660 (HSLp660) and 563 (HSLp563) in different adipose tissue depots as influenced by time and postpartum diet in dairy cows. Biopsy samples were obtained from s.c. (SCAT) and retroperitoneal (RPAT) adipose tissues of 20 Holstein cows 21 d prepartum, and 1 and 21 d postpartum. After d 1 postpartum, cows were randomly assigned to 2 groups (n=10). Groups received diets with either a concentrate-to-roughage ratio on a dry matter basis of 30:70% (low-concentrate, LC, group) or 60:40% (high-concentrate group), fed until the third biopsy sampling 21 d postpartum. Dry matter intake, milk yield, and milk composition were recorded. Blood samples were taken weekly, starting 21 d prepartum and analyzed for nonesterified fatty acids, ß-hydroxybutyrate (BHBA), glucose, and insulin. Protein expression of HSL and its extent of phosphorylation in adipose tissue were measured by semiquantitative Western blotting. Total HSL expression was lower in both adipose tissues 1 d after calving compared with prepartum sampling (SCAT: 4.10±0.5 vs. 2.4±0.3; RPAT: 11.1±1.3 vs. 6.6±1.1). Phosphorylation at Ser 660 was higher 21 d postpartum compared with 21 d prepartum in RPAT (2.9±0.3 vs. 4.6±0.6). Phosphorylation at Ser 563 was higher 21 d postpartum than 21 d prepartum in SCAT (0.6±0.1 vs. 3.9±1.1), and in RPAT a difference was observed between 21 d prepartum and 1 d postpartum (1.0±0.1 vs. 3.3. ± 0.6). On d 21 postpartum, the LC group showed a lower extent of Ser 563 phosphorylation in RPAT (3.9±0.8 vs.10.0±1.9) and a higher concentration of serum BHBA (0.77±0.05 vs. 0.47±0.11) than did the high-concentrate group. An inhibitory influence of higher BHBA concentrations on HSL phosphorylation in the LC group could be a possible explanation. On comparing RPAT to SCAT, HSL expression and the extent of Ser 660 and 563 phosphorylation was higher in RPAT at 21 d prepartum (HSL: 4.1±0.5 vs. 11.1±1.2; HSLp660 1.3±0.2 vs. 2.9±0.3; HSLp563: 0.6±0.1 vs. 1.0±0.1). In conclusion, the postpartum feeding regimen influenced the phosphorylation pattern, especially in RPAT, implying a regulatory role for different phosphorylation sites in adaptive lipolysis of dairy cows. It is suggested that RPAT is more sensitive to periparturient challenges than is SCAT.

The effect of intravenous enalaprilat (MK-422) administration in patients with mild to moderate essential hypertension.

The antihypertensive effect of enalaprilat (MK-422), an intravenous (IV), nonsulfhydryl converting-enzyme inhibitor, was evaluated in a double-blind study of 14 patients with mild to moderate hypertension. The seven patients in the treatment group initially received IV enalaprilat 1.25 mg q6h for 24 hours. Thereafter, responding patients (diastolic blood pressure [BP] less than or equal to 95 mm Hg) continued receiving this dose q6h for an additional 24 hours, whereas nonresponding patients were increased to IV enalaprilat 5 mg q6h for another 24 hours. Baseline BP for enalaprilat was 161 +/- 5/107 +/- 2 mm Hg (+/- SEM), and for placebo it was 150 +/- 5/103 +/- 2 mm Hg. Within the first 60 minutes, a significant reduction in both systolic and diastolic BP was noted in the enalaprilat group (P less than .05), without significant changes occurring in the placebo group. Although there was a gradual decline in both systolic and diastolic BP throughout the 48-hour study period in the placebo group, systolic and diastolic BP reduction was greater in the enalaprilat group, reaching a maximal decrease of 133 +/- 3/87 +/- 3 mm Hg. Adverse side effects did not occur in any patient.

Clinical use of 24-hour ambulatory blood pressure monitoring.

Knowing a patient's continuous 24-hour blood pressure measurements can be useful to the nurse practitioner in evaluating the hypertensive patient. Recent studies have demonstrated end-organ damage to have a higher degree of correlation with 24-hour blood pressure readings than with more periodic blood pressure readings. Previously, 24-hour ambulatory blood pressure determinations were evaluated primarily in research settings. The ability to obtain these readings for diagnostic information applicable to clinical situations is becoming readily available. The historical background leading to the development of the ambulatory blood pressure monitors for clinical use is reviewed. The indications for obtaining 24-hour ambulatory pressures, interpretation of specific graphs and patient outcome are explained.