Recombinant human IFN-alpha inhibits cerebral malaria and reduces parasite burden in mice

Most C57BL/6 mice infected i.p. with Plasmodium berghei ANKA (PbA) die between 7 and 14 days with neurologic signs, and the remainder die later (>15 days) with severe anemia. Daily i.p. injections of a recombinant human IFN-alpha (active on mouse cells) prevented death by cerebral malaria (87% deaths in the control mice vs 6% in IFN-alpha-treated mice). The mechanisms of this IFN-alpha protective effect were multiple. IFN-alpha-treated, PbA-infected mice showed 1) a marked decrease in the number of PbA parasites in the blood mediated by IFN-gamma, 2) less sequestered parasites in cerebral vessels, 3) reduced up-regulation of ICAM-1 expression in brain endothelial cells, 4) milder rise of blood levels of TNF, 5) increased levels of IFN-gamma in the blood resulting from an increased production by splenic CD8+ T cells, and 6) fewer leukocytes (especially CD8+ T cells) sequestered in cerebral vessels. On the other hand, IFN-alpha treatment did not affect the marked anemia observed in PbA-infected mice. Survival time in IFN-alpha-treated mice was further increased by performing three blood transfusions over consecutive days.     

Triaminoguanidine derivatives as supports to construct magnetic assemblies: Synthesis and characterization of trinuclear nickel(II) complexes

The synthesis of two nickel(II) complexes based on a central bridging triaminoguanidine scaffold and a capping ligand per metal ion is reported. When 2,2′-bipyridine (bipy) is utilized as co-ligand the complex [Ni₃L^Br(bipy)₃(H₂O)₃]NO₃ · 9H₂O · 1.5DMF (1) is obtained which crystallizes in the hexagonal space group P6₃/m. Complex 1 shows an interesting supramolecular structure pattern with alternating hydrophilic and hydrophobic layers characterized by extensive hydrogen-bonding and π-π-stacking, respectively. With 2,4,6-(2-pyridyl)-1,3,5-triazine (tptz) as capping ligand, complex [Ni₃L^Br(tptz)₃]ClO₄ · 7H₂O · 1.5DMF (2) is obtained. The magnetic susceptibility data can be fitted using an equilateral triangle model () with an isotropic coupling constant of J=-31.0±0.6 for 1 and for 2.     

The antitumor effects of interferon: a personal history

Early experiments showed that administration of mouse interferon preparations inhibited the development of viral-induced or spontaneous viral associated leukemias in mice. Interferon alpha/beta was also shown to inhibit the growth of transplantable tumors of different origins in all strains of mice tested. The finding that interferon alpha/beta inhibited the growth of sublines of tumors selected for resistance to interferon alpha/beta indicated the role of interferon induced host mechanisms in the antitumor effects observed. The different host antitumor mechanisms and especially the interaction of interferon alpha/beta with the immune system have been briefly discussed. Injection of mice with a neutralizing antibody to interferon alpha/beta demonstrated the essential role of endogenous interferon alpha/beta in the defense of the mouse against the development of syngeneic, allogeneic and xenogeneic tumors.     

Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated blood is the aorta. The aorta may become affected in diseases such as atherosclerosis and aneurysm. Researchers investigating these diseases would benefit from direct observation of the aorta to characterize disease progression as well as to evaluate efficacy of potential therapeutics. The goal of this protocol is to describe proper isolation and excision of the aorta to aid investigators researching cardiovascular disease. Isolation and excision of the aorta allows investigators to look at gross morphometric changes as wells as allowing them to preserve and stain the tissue to look at histologic changes if desired. The aorta may be used for molecular studies to evaluate protein and gene expression to discover targets of interest and mechanisms of action. This technique is superior to imaging modalities as they have inherent limitations in technology and cost. Additionally, primary isolated cells from a freshly isolated and excised aorta can allowing researchers to perform further in situ and in vitro assays. The isolation and excision of the aorta has the limitation of having to sacrifice the animal however, in this case the benefits outweigh the harm as it is the most versatile technique in the study of aortic disease.     

Identification of robust hypoxia biomarker candidates from fin of medaka (Oryzias latipes)

Aquatic hypoxia caused by organic pollution and eutrophication is a pressing worldwide water pollution problem. Better methods for monitoring oxygen levels are needed to assist efforts to maintain and protect the health of natural aquatic environments. In this project, we used a Japanese ricefish (medaka, Oryzias latipes) 8K oligonucleotide array as a platform to identify potential hypoxic biomarkers in different organs (fin, gill, liver and brain) upon exposure to hypoxia. The microarray results were validated by qRT-PCR employing a subset of candidate biomarkers. Interestingly, the largest number and most significant of hypoxia responding array features were detected in hypoxia exposed fin tissues. We identified 173 array features that exhibited a significant response (over 2 fold change in expression) upon exposure to hypoxic conditions and validated a subset of these by quantitative RT-PCR. These gene targets were subjected to annotation and gene ontology mining. Positively identifiable gene targets that may be useful for development of a rapid and accurate biomarker test using fin clips are discussed in relation to previous reports on hypoxia responsive genes.     

Polyostotic fibrous dysplasia in a cynomolgus Macaque (Macaca fascicularis)

A 2.3-y-old female cynomolgus macaque (Macaca fascicularis) presented with a broken right tibia and fibula. Radiographs showed multiple cyst-like defects in all long bones. We suspected that both fractures were pathologic because they occurred through these defects. Ultrasonography, MRI, and dual X-ray absorptiometry revealed that the defects were filled with soft tissue. Grossly, the bones were abnormal in shape, and a gelatinous material filled the defects and the surrounding marrow cavity. Histologically, the gelatinous material was composed of fibrin and cartilage; few normal bone cells were seen. Genetic testing revealed extra material on the short arm of chromosome 8 in all tissues examined, but no copy number alterations of likely clinical significance were observed, and no abnormalities were found that were unique to the lesions. In light of the clinical signs and radiographic and pathologic findings, polyostotic fibrous dysplasia was diagnosed. This report represents the first documented case of fibrous dysplasia in a cynomolgus macaque.     

Wet meadows of the alliance <Emphasis Type="Italic">Molinion</Emphasis> and their environmental gradients in Slovenia

The main objective of this study was to examine the relationships between wet meadow plant communities of Molinon alliance and their environmental conditions in Slovenia. The ecology of these communities was analysed in detail. The study provides the data on the vegetation and environmental parameters, the significance of parameters for the plant species composition, most important environmental gradients and differences between plant communities. In all plots the vegetation was recorded and soil parameters were analysed (pH, plant-available P and K, N^t, organic C, C/N ratio, exchangeable Ca²⁺, Mg²⁺, K⁺, Na⁺, H⁺, electrical conductivity, base saturation). Other conditions were also considered (e.g., mean annual temperature and precipitation, humidity index, mean Ellenberg moisture and nutrient value) to test possible correlations as well. Vegetation was classified by means of multivariate cluster analysis, while vegetation-site relationships were examined with direct gradient analysis (CCA). Six associations from the Molinon alliance (Selino-Molinietum, Plantagini altissimae-Molinietum, Carici davallianae-Molinietum, Gentiano-Molinietum litoralis, Junco conglomerati-Betonicetum and Sanguisorbo-Festucetum commutatae) were identified and analysed. Soil reaction was identified as most significant environmental parameter explaining the variation of the studied vegetation. There are several statistically significant differences in site conditions between the communities (pH, moisture, nutrient status, Ca²⁺). The studied associations represent clearly defined ecological units.     

Endothelial expression of beta1 integrin is required for embryonic vascular patterning and postnatal vascular remodeling

The largest subgroup of integrins is that containing the β1 subunit. β1 integrins have been implicated in a wide array of biological processes ranging from adhesion to cell growth, organogenesis, and mechanotransduction. Global deletion of β1 integrin expression results in embryonic death at ca. embryonic day 5 (E5), a developmental time point too early to determine the effects of this integrin on vascular development. To elucidate the specific role of β1 integrin in the vasculature, we conditionally deleted the β1 gene in the endothelium. Homozygous deletion of β1 integrins in the endothelium resulted in failure of normal vascular patterning, severe fetal growth retardation, and embryonic death at E9.5 to 10, although there were no overt effects on vasculogenesis. Heterozygous endothelial β1 gene deletion did not diminish fetal or postnatal survival, but it reduced β1 subunit expression in endothelial cells from adult mice by approximately 40%. These mice demonstrated abnormal vascular remodeling in response to experimentally altered in vivo blood flow and diminished vascularization in healing wounds. These data demonstrate that endothelial expression of β1 integrin is required for developmental vascular patterning and that endothelial β1 gene dosing has significant functional effects on vascular remodeling in the adult. Understanding how β1 integrin expression is modulated may have significant clinical importance.