In vivo degradation behavior of photo-cross-linked star-poly(epsilon-caprolactone-co-D,L-lactide) elastomers

We have recently reported on the preparation of biodegradable elastomers through photo-cross-linking acrylated star-poly(epsilon-caprolactone-co-D,L-lactide). In this paper we assess the change in their physical properties during in vivo degradation in rats after subcutaneous implantation over a 12 week period. These parameter changes were compared to those observed in vitro. Two different cross-link densities were examined, representing the range from a high Young's modulus to a low Young modulus. Elastomers having a high cross-link density exhibited degradation behavior consistent with a surface erosion mechanism, and degraded at the same rate in vivo as observed in vitro. Young's modulus and the stress at break of these elastomers decreased linearly with the degradation time, while the strain at break decreased slowly. Elastomers having a low cross-link density exhibited a degradation mechanism consistent with bulk erosion. Young's modulus and the stress at break of these elastomers decreased slowly initially, followed by a marked increase in mechanical strength loss after 4 weeks. The elastomers were well tolerated by the rats over the 12 week period in vivo.     

Continuous monitoring of predator control operations at landscape scale

Summary   Exotic predators are considered pests to wildlife and agriculture, requiring predator-control programs. Effective monitoring of predator-control operations is essential to justify their considerable cost, but often impossible in practice. The difficulties are especially severe if the target species is small and wide-ranging, and the area to be protected is inaccessible and/or extensive. A convenient model predator of this type, the feral Ferret ( Mustela furo ), is subject to control on farmland in New Zealand. We monitored the distribution of Ferrets over 2400 ha in the central North Island, before and throughout a standard control operation by professional trappers. We used 24 units of a new automated monitoring device, the Scentinel, set in a grid at 1 per 100 ha. Over 11 weeks (11 February to 29 April 2005, 1718 trap nights), we recorded 1559 visits by small mammals, including 198 by Ferrets. By the end of the 4th week, Ferrets had been detected at 17 of 24 sites. Removal of Ferrets from the study area by contractors began during the 5th week, and was reflected in significant declines in the number of Ferret visits recorded per day ( P  = 0.008) and the number of sites visited ( P  = 0.021). Analysis of our extensive repeat-survey data by site-occupancy methods confirmed these trends in greater detail, while also allowing for variation in detectability.     

Mechanical properties of individual focal adhesions probed with a magnetic microneedle

A permanent magnetic microneedle was developed to apply tensional forces to integrin receptors via ligand-coated magnetic microbeads while optically analyzing the mechanical properties of individual focal adhesions. Force application (130 pN for 3 s) through activated beta1 integrins produced less bead displacement than when unligated integrins were stressed. This strengthening response differed markedly on a bead-by-bead basis, correlated directly with local focal adhesion assembly, and was similar when analyzed at 4 degrees C, indicating that it was due to passive material properties of the cell. Viscoelastic analysis clarified that recruitment of focal adhesion proteins increased the local elastic stiffness of the adhesion complex without changing its viscous behavior. These data indicate that individual focal adhesions exhibit distinct mechanical properties that depend upon local focal adhesion assembly, and that these local variations in micromechanics can be detected and analyzed within living cells using the permanent magnetic microneedle technique.     

Ultrastructural evidence of autoinfection in the gills of Atlantic cod Gadus morhua infected with Loma sp. (phylum Microsporidia)

Infection by a microsporidian of the genus Loma was found in gills of cod Gadus morhua. Xenomas contained parasites in multiple stages of development. Some spores looked empty and had everted polar tubes, which were either straight or coiled. These polar tubes were scattered throughout the xenoma cytoplasm, and some of them pierced the plasma membrane. Those outside of the xenoma penetrated neighboring cells, including blood cells. These observations suggest that a mechanism of autoinfection could occur in blood cells and gill tissue, perpetuating the disease in the host.     

Activated human dendritic cells express inducible cyclo-oxygenase and synthesize prostaglandin E2 but not prostaglandin D2

Prostaglandins (PG) are well known lipid mediators with important immunoregulatory properties. While exogenous PGE2 has the ability to modulate the function and maturation of antigen presenting cells, such as dendritic cells (DC), it is not clear whether human DC have the capacity to synthesize PGE2 and other prostaglandins themselves. We therefore examined the expression of inducible cyclo-oxygenase (COX-2) by monocyte derived DC and the production of PGE2 and PGD2. Both monocyte derived DC and freshly isolated blood myeloid DC expressed little COX-2 constitutively, though COX-2 expression was rapidly but transiently upregulated in response to lipopolysaccharide stimulation. COX-2 mRNA was detectable within 1 h of LPS exposure, peaked at 4-6 h, and rapidly declined thereafter. COX-2 expression was accompanied by DC synthesis of PGE2, with peak levels present at 6-18 h post-stimulation. In contrast, PGD2 synthesis was not detected at any time point. When DC were activated with LPS in the presence of nimesulide, a COX-2 selective inhibitor, IL-10 synthesis was inhibited, indicating that endogenous prostaglandins regulate DC cytokine production. PGE2 production by DC may therefore modulate DC and T-cell function, thereby shaping the character of the immune response.     

New hominin first metatarsal (SK 1813) from Swartkrans

A recently recognized hominin hallucal metatarsal, SK 1813, from Swartkrans bears a suite of primitive and derived traits. Comparisons with extant apes, modern humans, SKX 5017, and Stw 562 reveals similar morphology in all three fossils and that these early hominins, while bipedal, possessed a unique toe-off mechanism. The implications of this are that both primitive and derived traits must be used to establish the total biomechanical pattern.     

Longitudinal analysis of deciduous tooth emergence: III. Sexual dimorphism in Bangladeshi, Guatemalan, Japanese, and Javanese children

Previous studies, mostly in European populations, found sex differences in the pattern of deciduous tooth emergence. Most studies find that the anterior dentition in males is precocial relative to the female dentition, and the pattern reverses so that females lead males in the emergence of the posterior deciduous dentition. Less is known about sex differences in the dental development and emergence of non-European populations. Here we examine the pattern of sex differences in deciduous tooth emergence in Japanese, Javanese, Guatemalan, and Bangladeshi children. The data come from four longitudinal or mixed longitudinal studies using similar study protocols. Survival analysis was used to estimate parameters of a log-normal distribution of emergence for each of the 10 teeth of the left dentition, and sexual dimorphism was assessed by sex-specific differences in mean emergence times and by Bennett's index. The results support the pattern of developmental cross-over observed in other populations. We conclude that there is little evidence to support the hypothesis of Tanguay et al. ([1984] J. Dent. Res. 63:65-68) that ethnic factors mediate sex differences in the emergence of deciduous teeth.     

Rabbit aorta converts 15-HPETE to trihydroxyeicosatrienoic acids: potential role of cytochrome P450

Previous work showed that rabbit aorta metabolizes arachidonic acid via 15-lipoxygenase to 15-hydroperoxyeicosatetraenoic acid (15-HPETE), which undergoes an enzymatic rearrangement to 11-hydroxy-14,15-epoxyeicosatrienoic acid (11-H-14,15-EETA) and 15-hydroxy-11,12-epoxyeicosatrienoic acid (15-H-11,12-EETA). Hydrolysis of the epoxy group results in the formation of 11,14,15- and 11,12,15-trihydroxyeicosatrienoic acids (THETAs). Endothelial cells have several heme-containing enzymes including cytochromes P450 (CYP), nitric oxide synthase (eNOS), and prostacyclin (PGI(2)) synthase that catalyze the rearrangement of 15-HPETE to HEETAs. Incubation of arachidonic acid and 15-lipoxygenase, or 15-HPETE with rabbit aortic microsomes or rat liver microsomes, a rich source of CYP, resulted in the formation of a product that comigrated with THETAs and HEETAs on HPLC. Immunoblot analysis showed the presence of CYP2C8 and CYP2J2 in aortic tissue and when CYP2J2 or CYP2C8 was incubated with arachidonic acid and 15-lipoxygenase, the major products were 11,12,15- and 11,14,15-THETAs. Incubation of purified hematin, CYP2C11, eNOS or PGI(2) synthase enzymes with arachidonic acid and 15-lipoxygenase produced a different pattern of metabolites from rabbit aortic microsomes. Clotrimazole, a non-specific CYP inhibitor, and ebastine and terfenadone, specific CYP2J2 inhibitors, blocked the ability of aortic microsomes to produce THETAs while specific inhibitors of PGI(2) synthase, eNOS or CYP2C8/2C9 had no effect on THETA production. We suggest that a CYP, possibly CYP2J2, may function as the hydroperoxide isomerase converting 15-HPETE to HEETAs in rabbit vascular tissue. Further hydrolysis of the epoxy group of the HEETAs results in the formation of 11,12,15- and 11,14,15-THETAs. The HEETAs and THETAs are both vasodilators and may function as important regulators of vascular tone.     

Processing and localization of ADAMTS-1 and proteolytic cleavage of versican during cumulus matrix expansion and ovulation

ADAMTS-1 (a disintegrin and metalloprotease with thrombospondin motifs-1) is a member of the ADAMTS family of metalloproteases which, together with ADAMTS-4 and ADAMTS-5, has been shown to degrade members of the lectican family of proteoglycans. ADAMTS-1 mRNA is induced in granulosa cells of periovulatory follicles by the luteinizing hormone surge through a progesterone receptor-dependent mechanism. Female progesterone receptor knockout (PRKO) mice are infertile primarily due to ovulatory failure and lack the normal periovulatory induction of ADAMTS-1 mRNA. We therefore investigated the protein localization and function of ADAMTS-1 in ovulating ovaries. Antibodies against two specific peptide regions, the pro-domain and the metalloprotease domain of ADAMTS-1, were generated. Pro-ADAMTS-1 of 110 kDa was identified in mural granulosa cells and appears localized to cytoplasmic secretory vesicles. The mature (85-kDa pro-domain truncated) form accumulated in the extracellular matrix of the cumulus oocyte complex (COC) during the process of matrix expansion. Each form of ADAMTS-1 protein increased >10-fold after the ovulatory luteinizing hormone surge in wild-type but not PRKO mice. Versican is also localized selectively to the ovulating COC matrix and was found to be cleaved yielding a 70-kDa N-terminal fragment immunopositive for the neoepitope DPEAAE generated by ADAMTS-1 and ADAMTS-4 protease activity. This extracellular processing of versican was reduced in ADAMTS-1-deficient PRKO mouse ovaries. These observations suggest that one function of ADAMTS-1 in ovulation is to cleave versican in the expanded COC matrix and that the anovulatory phenotype of PRKO mice is at least partially due to loss of this function.