Strain-based estimation of time-dependent transmural myocardial architecture in the ovine heart

Left ventricular myofibers are connected by an extensive extracellular collagen matrix to form myolaminar sheets. Histological cardiac tissue studies have previously observed a pleated transmural distribution of sheets in the ovine heart, alternating sign of the sheet angle from epicardium to endocardium. The present study investigated temporal variations in myocardial fiber and sheet architecture during the cardiac cycle. End-diastolic histological measurements made at subepicardium, midwall, and subendocardium at an anterior-basal and a lateral-equatorial region of the ovine heart, combined with transmural myocardial Lagrangian strains, showed that the sheet angle but not the fiber angle varied temporally throughout the cardiac cycle. The magnitude of the sheet angle decreased during systole at all transmural depths at the anterior-basal site and at midwall and subendocardium depths at the lateral-equatorial site, making the sheets more parallel to the radial axis. These results support a previously suggested accordion-like wall-thickening mechanism of the myocardial sheets.     

Animal Housing and Welfare: Effects of Housing Conditions on Body Weight and Cortisol in a Medium-Sized Rodent (Cavia aperea)

Rodents are the most abundant experimental nonhuman animals and are commonly studied under standard laboratory housing conditions. As housing conditions affect animals' physiology and behavior, this study investigated the effects of indoor and outdoor housing conditions on body weight and cortisol level of wild cavies, Cavia aperea. The changing housing condition strongly influenced both parameters, which are commonly used as indicators for animal welfare. The transfer from outdoor to indoor enclosures resulted in a body-weight loss of about 8%. In contrast, animals kept indoors showed a substantial weight gain of about 12% when they were transferred outdoors. These effects were reversible. To substantiate a connection between body-weight changes and the health states of the animals, blood basal cortisol concentrations were measured. Animals kept outdoors had significantly lower cortisol levels than did animals kept indoors. These results imply that indoor conditions have a direct effect on the animals' states. The physiological and metabolic consequences as well as potential welfare aspects should be taken into account when planning experimental work, especially on nondomestic animals.     

Two waves of the nuclear phospholipase C activity in serum-stimulated HL-60 cells during G(1) phase of the cell cycle

Phosphatidylinositol-specific phospholipase C (PI-PLC) is activated in cell nuclei during the cell cycle progression. We have previously demonstrated two peaks of an increase in the nuclear PI-PLC activities in nocodazole-synchronized HL-60 cells. In this study, the activity of nuclear PI-PLC was investigated in serum-stimulated HL-60 cells. In serum-starved HL-60 cells, two peaks of the activity of nuclear PI-PLC were detected at 30 min and 11 h after the re-addition of serum with no parallel increase in PLC activity in cytosol, postnuclear membranes or total cell lysates. An increase in the serine phosphorylation of b splicing variant of PI-PLCbeta(1) was detected with no change in the amount of PI-PLCbeta(1b) in nuclei isolated at 30 min and 11 h after the addition of serum. PI-PLC inhibitor ET-18-OCH(3) and MEK inhibitor PD 98059 completely abolished serum-mediated increase at both time-points. The addition of inhibitors either immediately or 6 h after the addition of serum had inhibitory effects on the number of cells entering S phase. These results demonstrate that two waves of nuclear PI-PLCbeta(1b) activity occur in serum-stimulated cells during G(1) phase of the cell cycle and that the later increase in the PLC activity is equally important for the progression into the S phase.     

Synthesis and biological evaluation of a series of A,B-ring modified 16,17-secoandrostane derivatives

The starting compound for the synthesis of 16,17-secoandrostane derivatives with the 4-en-3-on, 1,4-dien-3-on, 4,6-dien-3-on, and 1,4,6-trien-3-on systems was 3β-hydroxy-17-methyl-16,17-secoandrost-5-en-16-nitrile-17-one (1), the Oppenauer oxidation of which yielded the corresponding 4-en-3-one derivative 2. Dehydrogenation of compound 2 with the aid of 2,3,5,6-tetrachloro-1,4-benzoquinone (chloranil) gave the three products: 17-methyl-16,17-secoandrosta-1,4-dien-3,17-dione-16-nitrile (3), 17-methyl-16,17-secoandrosta-4,6-dien-3,17-dione-16-nitrile (4), and 17-methyl-16,17-secoandrosta-1,4,6-trien-3,17-dione-16-nitrile (5). On the other hand, epoxidation of compound 2 resulted in a mixture of α and β isomers of 4,5-epoxy-17-methyl-16,17-secoandrosta-3,17-dione-16-nitrile (6 and 7). Opening of the oxirane rings of the mixture of 6 and 7 by the action of formic acid yielded the 4-hydroxy-4-en derivative 8. Antiaromatase activity and in vitro cytotoxicity against three tumor cell lines (human breast adenocarcinoma ER+, MCF-7, human breast adenocarcinoma ER−, MDA-MB-231, and prostate cancer PC3) of selected compounds were evaluated. Compound 2 exhibited a relatively strong inhibition of aromatase and extremely potent cytotoxicity against PC3 cells. Compound 8 showed satisfactory cytotoxicity against MCF-7 cells.     

Brazilian mothers' beliefs, attitudes and practices related to child weight status and early feeding within the context of nutrition transition

With the rapid pace of the nutrition transition worldwide, understanding influences of child feeding practices within a context characterized by the co-existence of overweight and undernutrition in the same population has increasing importance. This qualitative study describes Brazilian mothers' child feeding practices and their perceptions of their association with child weight status and explores the role of socioeconomic, cultural and organizational factors on these relationships. Forty-one women enrolled in the Family Health/Community Health Workers Programme were selected from rural, urban, coastal and indigenous areas in Ceara State, north-east Brazil, to participate in four focus group discussions. Content analysis identified fourteen emergent themes showing mothers' child feeding practices in this setting were influenced by economic resources, mothers' immediate social support networks (e.g. neighbours and family members) and participation in nutrition assistance programmes. Child malnutrition was the most common nutritional concern; nevertheless, mothers were aware of the negative health consequences of obesity but misunderstood its causes (e.g. foods filled with fat would make a person fat; others thought that birth control pills and stimulants given to children were causes of obesity); several reported their own struggles with overweight. Food assistance programmes emerged as an important influence on children's dietary adequacy, especially among mothers describing dire economic situations. The findings have implications for targeting food assistance as well as health and nutrition education strategies in low-income families undergoing the nutrition transition in north-east Brazil.     

Factors influencing the appearance of spider (Araneae) and beetle (Coleoptera) assemblages in nests of great reed warbler Acrocephalus arundinaceus

The appearance of spider (Araneae) and beetle (Coleoptera) assemblages found in nests of great reed warbler Acrocephalus arundinaceus was studied, firstly to investigate breeding success and the amount of precipitation as potential factors which might affect the abundance and species richness of both groups. In addition, we compared the diversity of spider and beetle assemblages between nests found in different reed habitats, and considered the position of nests (above water or dry ground). In this study we selected five different randomly chosen reed habitats: two mining ponds, two small canals and one large canal. Great Reed Warbler nests were collected either shortly after fledging, or after the clutch had failed. Altogether, 12 species of spider and 19 species of beetle were collected. In both groups there was no significant difference in abundance between successful, lost and cuckoo-parasitized nests; however, there was a significant difference in species richness between the three nest categories in spider assemblages, which was not the case in beetle assemblages. The amount of precipitation did not affect beetle or spider abundance; only the species richness of spiders showed significant growth. Furthermore, we found no significant relationship between vegetation cover and the species richness and abundance of spiders and beetles. The diversity of both groups differed significantly according to reed habitat: beetle assemblages were most diverse by the large canal and spiders at the mining ponds.     

G-protein β3 subunit gene C825T polymorphism in patients with vesico-ureteric reflux

The C825T polymorphism in the GNB3 gene encoding a β3 subunit from heterotrimeric G-proteins correlates strongly with the variation in activity of the G-proteins. It has so far been associated with a variety of medical conditions, but has not been tested for association with vesico-ureteric reflux (VUR). Primary VUR is a condition of genetic origin that appears to be inherited in an autosomal dominant mode, but with reduced penetrance. The constitutional change in G-protein-mediated cell signaling associated with the C825T polymorphism might be one of the factors that participate in the development of VUR by modifying the effect of still unknown mutated gene(s). A significant difference in genotype frequencies (χ² = 7.38, P = 0.025, df = 2) was observed between patients with primary VUR (33 CC homozygotes, 40 CT heterozygotes, 12 TT homozygotes) and healthy controls with no medical record of reflux (114 CC homozygotes, 88 CT heterozygotes, 18 TT homozygotes). This result suggests that the C825T polymorphism of the GNB3 gene might be associated with the development of VUR.     

Activity cage as a method to analyze functional recovery after sciatic nerve injury in mice

The aim of this paper is to show the activity cage as a viable method for tracking functional nerve recovery. The activity cage measures spontaneous coordinate activity, meaning movement in either the horizontal or vertical plane, of experimental animals within a specified amount of time. This uses a minimum of researcher time conducting functional testing to determine functional recovery of the nerve. Using microsurgical forceps, a crush injury was inflicted unilaterally, on the left side, upon the 4-month-old C3H mice creating a very high degree of pressure for 6 s upon the exposed sciatic nerve. The locomotion function of the mice was evaluated using the activity cage preoperatively, 1, 7, 14, 21, and 28 days after the surgical procedure. We found that using the activity cage functional recovery occurred by 14 days after nerve crush injury. It was also shown that, coinciding with functional recovery, immunohistochemistry changes for GD1a and nNOS appeared at the level of L4, where the sciatic nerve joins the spinal column. GD1a and nNOS have both been linked to regenerative processes in mammalian nervous systems.     

Nanosized bilayer disks: attractive model membranes for drug partition studies

Stable nanosized bilayer disks were prepared from either 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and cholesterol, or lipid mixtures with a composition reflecting that of the porcine brush border membrane. Two different polyethylene glycol (PEG)-grafted lipids, the negatively charged 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy (polyethylene glycol)-5000] (DSPE-PEG(5000)) and the neutral N-palmitoyl-sphingosine-1-[succinyl (methoxy (polyethylene glycol) 5000] (Ceramide-PEG(5000)), were used to stabilize the disks. The disks were employed as model membranes in drug partition studies based on a fast chromatography method. Results show that the lipid composition, as well as the choice of PEG-lipid, have an important influence on the partition behavior of charged drugs. Comparative studies using multilamellar liposomes indicate that bilayer disks have the potential to generate more accurate partition data than do liposomes. Further, initial investigations using bacteriorhodopsin suggest that membrane proteins can be reconstituted into the bilayer disks. This fact further strengthens the potential of the bilayer disk as an attractive model membrane.