Scanning X‐ray microdiffraction of decellularized pericardium tissue at increasing glucose concentration

Blood glucose supplies energy to cells and is critical for the human brain. Glycation of collagen, the nonenzymatic formation of glucose‐bridges, relates to diseases of aging populations and diabetics. This chemical reaction, together with its biomechanical effects, has been well studied employing animal models. However, the direct impact of glycation on collagen nano‐structure is largely overlooked, and there is a lack of ex vivo model systems. Here, we present the impact of glucose on collagen nanostructure in a model system based on abundantly available connective tissue of farm animals. By combining ex vivo small and wide‐angle X‐ray scattering (SAXS/WAXS) imaging, we characterize intra‐ and inter‐molecular parameters of collagen in decellularized bovine pericardium with picometer precision. We observe three distinct regimes according to glucose concentration. Such a study opens new avenues for inspecting the effects of diabetes mellitus on connective tissues and the influence of therapies on the resulting secondary disorders.      

Curcumin protects cardiac cells against ischemia-reperfusion injury: effects on oxidative stress, NF-kappaB, and JNK pathways

In this study we explored the effects of curcumin in cardiac cells subjected to a protocol simulating ischemia-reperfusion (IR). Curcumin (10 microM) was administered before ischemia (pretreatment) or at the moment of reperfusion (posttreatment) and its effects were compared to those produced by a reference antioxidant (Trolox) with an equal antioxidant capacity. IR cardiac cells showed clear signs of oxidative stress, impaired mitochondrial activity, and a marked development of both necrotic and apoptotic processes; at the same time, increases in NF-kappaB nuclear translocation and JNK phosphorylation were observed. Curcumin pretreatment was revealed to be the most effective in attenuating all these modifications and, in particular, in reducing the death of IR cells. This confirms that the protective effect of curcumin is not related simply to its antioxidant properties but involves other mechanisms, notably interactions in the NF-kappaB and JNK pathways. These findings suggest that curcumin administration, in particular before the hypoxic challenge, represents a promising approach to protecting cardiac cells against IR injury. In this scenario our results point out the importance of the chronology for the outcome of the treatment and provide a differential valuation of the degree of protection that curcumin can exert by its antioxidant activity or by other mechanisms.     

Kinematics of the human ankle complex in passive flexion; a single degree of freedom system

The restoration of original range and pattern of motion is the primary goal of joint replacement and ligament reconstruction. The objective of the present work is to investigate whether or not a preferred path of joint motion at the intact human ankle complex is exhibited during passive flexion. A rig was built to move the ankle complex through its range of flexion while applying only the minimum necessary load to drive ankle flexion. Joint motion was constrained only by the articular surfaces and the ligaments. The movements of the calcaneus, talus and fibula relative to the stationary tibia in seven cadaveric specimens were tracked with a stereophotogrammetric system. It was shown that the calcaneus follows a unique path of unresisted coupled motion relative to the tibia and that most of the motion occurred at the ankle, with little motion at the subtalar level. The calcaneofibular and the tibiocalcaneal ligaments showed near-isometric pattern of rotations. All specimens showed motion of the axis of rotation relative to the bones. Deviations from the unique path due to the application of load involved mostly subtalar motion and were resisted. The ankle complex exhibits one degree of unresisted freedom, the ankle behaving as a single degree of freedom mechanism and the subtalar as a flexible structure. We deduced that the calcaneofibular and tibiocalcaneal ligaments together with the articular surfaces guide ankle passive motion, other ligaments limit but do not guide motion.     

Ecological niche modeling and geographical distribution of pollinator and plants: A case study of Peponapis fervens (Smith, 1879) (Eucerini: Apidae) and Cucurbita species (Cucurbitaceae)

The bees of the Peponapis genus (Eucerini, Apidae) have a Neotropical distribution with the center of species diversity located in Mexico and are specialized in Cucurbita plants, which have many species of economic importance, such as squashes and pumpkins. Peponapis fervens is the only species of the genus known from southern South America. The Cucurbita species occurring in the same area as P. fervens include four domesticated species (C. ficifolia, C. maxima maxima, C. moschata and C. pepo) and one non-domesticated species (Cucurbita maxima andreana). It was suggested that C. m. andreana was the original pollen source to P. fervens, and this bee expanded its geographical range due to the domestication of Cucurbita. The potential geographical areas of these species were determined and compared using ecological niche modeling that was performed with the computational system openModeller and GARP with best subsets algorithm. The climatic variables obtained through modeling were compared using Cluster Analysis. Results show that the potential areas of domesticated species practically spread all over South America. The potential area of P. fervens includes the areas of C. m. andreana but reaches a larger area, where the domesticated species of Cucurbita also occur. The Cluster Analysis shows a high climatic similarity between P. fervens and C. m. andreana. Nevertheless, P. fervens presents the ability to occupy areas with wider ranges of climatic variables and to exploit resources provided by domesticated species.     

Synthesis and opioid activity of N,N-dimethyl-Dmt-Tic-NH-CH(R)-R' analogues: acquisition of potent delta antagonism

N,N-Dimethylation of the H-Dmt-Tic-NH-CH(R)-R′ series of compounds produced no significant affect on the high δ-opioid receptor affinity (K_i=0.035–0.454 nM), but dramatically decreased that for the μ-opioid receptor. The effect of N-methylation was independent of the length of the linker (R); however, the bioactivities were affected by the chemical composition of the third aromatic group (R′): phenyl (Ph) (5′–8′) elicited a greater reduction in μ-affinity (40–70-fold) compared to analogues containing 1H-benzimidazole-2-yl (Bid) (9-fold). The major consequences of N,N-dimethylation on in vitro bioactivity were: (i) a loss of δ-agonism coupled with the appearance of potent δ antagonism (4′–7′) (pA₂=8.14–9.47), while 1 exhibited only a 160-fold decreased δ agonism (1′) and the δ antagonism of 8 enhanced >10-fold (pA₂=10.62, 8′); and (ii) a consistent loss of μ-affinity resulted in enhanced δ-opioid receptor selectivity. With the exception of compound 1′, the change in the hydrophobic environment at the N-terminus and formation of a tertiary amine by N,N-dimethylation in analogues of the Dmt-Tic pharmacophore produced potent δ-selective antagonists.     

Phylogeny and evolution of body mass in didelphid marsupials (Marsupialia: Didelphimorphia: Didelphidae)

Most extant New World marsupials belong in the Didelphidae, which comprises ca. 110 currently recognized species of opossums. Didelphids are small mammals with their mean body mass, at species level, ranging from ca. 7 g to 2.2 kg. The largest species belong in a single clade, while substantial variation remains scattered across the remaining groups. We seek out to explore the details of this mass variation in an evolutionary framework. To this end, we first reconstructed the phylogeny of didelphids based on an extensive, although fragmentary sample of sequences from ten genes. We recovered a fully resolved, highly robust phylogeny that tested and confirmed most previously reported groupings, providing a simultaneous depiction of phylogenetic relationships for 81 % of currently recognized species and all relevant supra-specific clades. As much as 69 % of total body mass variation in didelphids was explained by this phylogenetic hypothesis. Mapped on it, mass variation evolved as much as 6.8 kg of total changes, starting from a reconstructed ancestral body mass range of 22–33 g. No single, family-wide pattern was evident; in fact, the dominant pattern for mass variation was that of increases in body mass along a few successive branches, or phyletic giantism, followed by apomorphic nanism, i.e., decreases localized in single terminal branches. Phyletic trends indicated the persistence of gradual, directional changes along considerable spans of geological time and show that substantial variation of interest resides in this and perhaps most groups of small mammals.     

Chlorophyll fluorescence and photosynthetic response to light in 1-year-old needles during spring and early summer in <Emphasis Type="Italic">Pinus leucodermis</Emphasis>

Ten-year-old trees from four Italian populations of Pinus leucodermis (populations A, B, C and D), which were collected from different sites at different altitudes, were grown near Florence, Italy. Needle CO₂ gas exchange and chlorophyll fluorescence response to increasing light intensities were evaluated; gas exchange and chlorophyll fluorescence variation between April and July were also monitored. Populations A, B and C showed a similar photosynthetic response to increasing photosynthetic photon flux density (PPFD) intensities, while at various light intensities population D, which originated from the highest altitude, showed the highest photosynthetic rates. In this population photosynthesis was saturated at PPFDs higher than 900 µmol m^-2s^-1 and a slow decrease of effective photosystem II quantum yield and F'_V/F'_M in response to increasing PPFDs were found. The same trees also showed a faster recovery in photosynthesis from limitations induced by winter temperatures than the other three populations. This work showed that photosynthetic response to light in population D was different from the other populations; trees from this population were probably naturally selected to prevent photoinhibition due to excess light.     

IL-4 promotes the migration of circulating B cells to the spleen and increases splenic B cell survival

We report that IL-4 causes a redistribution of B cells and modestly increases B cell life span. Intravenous injection of a long-acting formulation of IL-4 induces increases in both spleen cell number and the percentage of splenic B cells. These effects are observed within 1 day of IL-4 administration and plateau after approximately 3 days if IL-4 treatment is continued. The increase in splenic B cell number is IL-4 dose dependent, CD4+ T cell independent, FcgammaRII/FcgammaRIII independent, and Stat6 independent. Decreases in the number of B cells in the blood and the percentage of mature B cells in the bone marrow, concomitant with the increase in splenic B cell number, suggest that redistribution of circulating B cells to the spleen is partially responsible for IL-4 induction of splenic B cell hyperplasia. Considerable reduction in the effect of 5 days of IL-4 treatment on splenic B cell number when B lymphopoiesis is blocked with anti-IL-7 mAb suggests that generation of new B cells is also involved in IL-4-induced splenic B cell hyperplasia. 5-Bromo-2'-deoxyuridine labeling experiments demonstrate that IL-4 modestly prolongs the life span of newly generated splenic B cells, and experiments that measure B cell HSA (CD24) expression as an indicator of B cell age suggest that IL-4 may also prolong the life span of mature splenic B cells. Thus, IL-4 increases splenic B cell number through two Stat6-independent effects: increased net migration of circulating B cells to the spleen and increased B cell life span. Both effects may promote Ab responses to a systemic Ag challenge.     

No Orcadian Rhythms of Serotoninergic, Alpha-, Beta-Adrenergic and Imipramine Binding Sites in Rat Brain Regions

B_max values of the specific binding of [³H]-WB 4101, [³H]-dihydroalprenolol, [³H]-spiperone and [³H]-imipramine to various rat brain regions were determined at 4 hr intervals over 24 hr under circadian conditions. No significant circadian rhythm of binding sites number was found for any receptor investigated in cerebral cortex, hypothalamus or brain stem. Some methodological issues are discussed.