Pathogenesis of ankylosing spondylitis: relation between HLA-B 27 and Klebsiella aerogenes

The Authors attested the evidence of a cross-reactivity among some Klebsiella antigens and HLA B 27 positive lymphocytes, suggesting the hypothesis that Klebsiella may be an initiating agent in ankylosing spondylitis (AS). The Authors determined also serum antibody response against Klebsiella antigens in AS patients. The results obtained are discussed.     

Dual role of parathyroid hormone in endothelial progenitor cells and marrow stromal mesenchymal stem cells

Hematopoietic stem cells derive regulatory information also from parathyroid hormone (PTH). To explore the possibility that PTH may have a role in regulation of other stem cells residing in bone marrow, such as mesenchymal stem cells (MSCs) and endothelial progenitor cells (EPCs) we assessed the effect of this hormone on the in vitro behavior of MSCs and EPCs. We evidenced that MSCs were much more responsive to PTH than EPCs. PTH increased the proliferation rate of MSCs with a diminution of senescence and apoptosis. Taken together, our results may suggest a protective effect of PTH on MSCs that reduces stress phenomena and preserve genome integrity. At the opposite, PTH did not modify the fate of EPCs in culture. J. Cell. Physiol. 222: 474–480, 2010. © 2009 Wiley‐Liss, Inc.     

Mitral re-repair and right coronary fistula closure: advantage of minimally invasive approach

A 51-year-old man developed severe mitral regurgitation 10 years after previous mitral valve repair; the echocardiographic images showed a remarkable eccentric jet toward posterior wall of left atrium associated with a high degree of pulmonary vein retrograde flow. The coronary arteriography pointed out no pathologic lesions but a coronary fistula from the proximal right coronary to the right atrium. The standard approach was avoided, and a right anterolateral minithoracotomy was chosen, providing an excellent view. Under cardiopulmonary bypass and mild hypothermia, the mitral valve was re-repaired, and a new ring was implanted. After aortic cross-clamp release, the right coronary fistula was closed through the right atrium. The postoperative course was uneventful, and the patient was discharged on the fourth postoperative day. In such a high-risk reintervention and concomitant procedure, we think that this different approach may represent a feasible and reliable alternative.     

Hemoglobin structure/function and globin-gene evolution in the Arctic fish Liparis tunicatus

The importance of the Arctic, in contributing to the knowledge of the overall ensemble of adaptive processes influencing the evolution of marine organisms, calls for investigations on molecular adaptations in Arctic fish. Unlike the vast majority of Antarctic Notothenioidei, several Arctic species display high hemoglobin multiplicity. The blood of four species, the spotted wolffish of the family Anarhichadidae and three Gadidae, contains three functionally distinct major components. Similar to many Antarctic notothenioids, Arctic Liparis tunicatus (suborder Cottoidei, family Liparidae) has one major hemoglobin (Hb 1) accompanied by a minor component (Hb 2). This paper reports the structural and functional characterisation of Hb 1 of L. tunicatus. This hemoglobin shows low oxygen affinity, and pronounced Bohr and Root effects. The amino-acid sequence of the beta chain displays an unusual substitution in NA2 (beta2) at the phosphate-binding site, and the replacement of Val E11 (beta67) with Ile. Similar to some Antarctic fish Hbs, electron paramagnetic resonance spectra reveal the formation of a ferric penta-coordinated species even at physiological pH. The amino-acid sequences have also been used to gain insight into the evolutionary history of globins of polar fish. L. tunicatus globins appear close to the notothenioid clades as predicted by teleostean phylogenies. Close phylogenetic relationships between Cottoidei and Notothenioidei, together with their life style, seem to be the main factor driving the globin-sequence evolution.     

An effective method for adenoviral-mediated delivery of small interfering RNA into mesenchymal stem cells

Mesenchymal stem cells (MSCs) promise as a main actor of cell-based therapeutic strategies, due to their intrinsic ability to differentiate along different mesenchymal cell lineages, able to repair the diseased or injured tissue in which they are localized. The application of MSCs in therapies requires an in depth knowledge of their biology and of the molecular mechanisms leading to MSC multilineage differentiation. The knockdown of target genes through small interfering RNA (siRNA) carried by adenoviruses (Ad) represents a valid tool for the study of the role of specific molecules in cell biology. Unfortunately, MSCs are poorly transfected by conventional Ad serotype 5 (Ad5) vectors. We set up a method to obtain a very efficient transduction of rat MSCs with low doses of unmodified Ad5, carrying the siRNA targeted against the mRNA coding for Rb2/p130 (Ad-siRNA-Rb2), which plays a fundamental role in cell differentiation. This method allowed a 95% transduction rate of Ad-siRNA in MSC, along with a siRNA-mediated 85% decrease of Rb2/p130 mRNA and a 70% decrease of Rb2/p130 protein 48 h after transduction with 50 multiplicities of infection (MOIs) of Ad5. The effect on Rb2/p130 protein persisted 15 days after transduction. Finally, Ad-siRNA did not compromise the viability of transduced MSCs neither induced any cell cycle modification. The effective Ad-siRNA-Rb2 we constructed, together with the efficient method of delivery in MSCs we set up, will allow an in depth analysis of the role of Rb2/p130 in MSC biology and multilineage differentiation.     

High-Yielding Enzymatic α-Glucosylation of Pyridoxine by Marine α-Glucosidase from Aplysia fasciata

We recently succeeded in the identification and purification of an interesting marine exo-α-glucosidase (EC 3.2.1.20) from the anaspidean mollusc Aplysia fasciata. The enzyme was characterized by good transglycosylation activity toward different acceptors using maltose as donor. High-yielding enzymatic α-glycosylation of pyridoxine using this marine enzyme is reported here; the reaction has been optimized, reaching 80% molar yield of products (pyridoxine monoglucosides 24 g/l; pyridoxine isomaltoside 35 g/l). High selectivity toward the 5′ position is observed for both monoglucoside and disaccharide formation. This is the first report describing the enzymatic production of pyridoxine isomaltoside.     

A dynamic culture platform enhances the efficiency of the 3D HUVEC-based tube formation assay

Cell-based in vitro biological models traditionally use monolayer cell cultures grown over plastic surfaces bathing in static media. Higher fidelity to a natural biological tissue is expected to result from growing the cells in a three-dimensional (3D) matrix. However, due to the decreased rate of diffusion inherent to increased distances within a tridimensional space, proper fluidic conditions are needed in this setting to better approximate a physiological environment. To this aim, we here propose a prototypal dynamic cell culture platform for the automatic medium replacement, via periodic perfusion flow, in a human umbilical vein endothelial cell (HUVECs) culture seeded in a Geltrex™ matrix. A state-of-the-art angiogenesis assay performed in these dynamic conditions showed sizable effects with respect to conventional static control cultures, with significantly enhanced pro-(dual antiplatelet therapy [DAPT]) and anti-(EDTA) angiogenic compound activity. In particular, dynamic culture conditions (a) enhance the 3D-organization of HUVECs into microtubule structure; (b) accelerate and improve endothelial tube formation by HUVECs in the presence of DAPT; (c) are able to completely revert the blocking effects of EDTA. These evidence emphasize the need of setting proper fluidic conditions for a better approximation of a physiological environment as an appropriate evolution of current cell culture paradigms.     

Occupancy,density, and activity patterns of a Critically Endangered leopard population on the Kawthoolei-Thailand border

Large carnivores have been largely extirpated from Southeast Asia due to deforestation, habitat fragmentation, and poaching. Estimating the density of endangered carnivore populations, and identifying relationships between species occupancy and both environmental and anthropogenic factors, is essential for effective conservation planning. Recently, the IUCN conservation status of the Indochinese leopard (Panthera pardus delacouri) was upgraded to “Critically Endangered.” We surveyed Kweekoh Wildlife Sanctuary in Kawthoolei, an area administered by the Karen ethnic group in eastern Myanmar, to quantify (1) leopard population density using spatially explicit mark-resight (SMR) models, (2) leopard occupancy as influenced by important ecological variables, and (3) potential differences in activity between melanistic and spotted leopard morphs. Leopard density was estimated to be 1.39 ± SE 0.22/100 km². Leopard occupancy (ψ = 0.43; 95% credible interval: 0.26–0.67) increased further from roads, at relatively higher elevations, and in areas with higher relative abundance of wild boar. Leopard activity was cathemeral, with higher activity during night hours, and significant overlap (Δ = 0.84; 95% confidence interval: 0.71–0.96) between melanistic and spotted morphs. However, melanistic leopards were more active during twilight hours than spotted individuals whose activity did not significantly vary throughout the day. Indochinese leopard density estimates in Kweekoh were among the lowest reported from Southeast Asia. Leopard occupancy was highest in the sanctuary's core areas, suggesting the presence of negative anthropogenic impacts along the sanctuary borders. We suggest our low density estimates warrant immediate and decisive conservation action, including better protection for leopards, their habitat, and their prey.