Pulmonary hemodynamics and lung water content during splanchnic ischemic shock

Pulmonary hemodynamics and lung water content were evaluated in open-chest dogs during splanchnic arterial occlusion (SAO) shock. Mean pulmonary arterial pressure [Ppa = 13.0 +/- 0.6 (SE) mmHg] and pulmonary venous pressure (4.1 +/- 0.2 mmHg) were measured by direct cannulation and the capillary pressure (Ppc = 9.0 +/- 0.6 mmHg) estimated by the double-occlusion technique. SAO shock did not produce a significant change in Ppa or Ppc despite a 90% decrease in cardiac output. An 18-fold increase in pulmonary vascular resistance occurred, and most of this increase (70%) was on the venous side of the circulation. No differences in lung water content between shocked and sham-operated dogs were observed. The effect of SAO shock was further evaluated in the isolated canine left lower lobe (LLL) perfused at constant flow and outflow pressure. The addition of venous blood from shock dogs to the LLL perfusion circuit caused a transient (10-15 min) increase in LLL arterial pressure (51%) that could be reversed rapidly with papaverine. In this preparation, shock blood produced either a predominantly arterioconstriction or a predominantly venoconstriction. These results indicate that both arterial and venous vasoactive agents are released during SAO shock. The consistently observed venoconstriction in the intact shocked lung suggests that other factors, in addition to circulating vasoactive agents, contribute to the pulmonary hemodynamic response of the open-chest shocked dog.     

Asymptomatic Cattle Naturally Infected with Mycobacterium bovis Present Exacerbated Tissue Pathology and Bacterial Dissemination

Rational discovery of novel immunodiagnostic and vaccine candidate antigens to control bovine tuberculosis (bTB) requires knowledge of disease immunopathogenesis. However, there remains a paucity of information on the Mycobacterium bovis-host immune interactions during the natural infection. Analysis of 247 naturally PPD+ M. bovis-infected cattle revealed that 92% (n = 228) of these animals were found to display no clinical signs, but presented severe as well as disseminated bTB-lesions at post-mortem examination. Moreover, dissemination of bTB-lesions positively correlated with both pathology severity score (Spearman r = 0.48; p<0.0001) and viable tissue bacterial loads (Spearman r = 0.58; p = 0.0001). Additionally, granuloma encapsulation negatively correlated with M. bovis growth as well as pathology severity, suggesting that encapsulation is an effective mechanism to control bacterial proliferation during natural infection. Moreover, multinucleated giant cell numbers were found to negatively correlate with bacterial counts (Spearman r = 0.25; p = 0.03) in lung granulomas. In contrast, neutrophil numbers in the granuloma were associated with increased M. bovis proliferation (Spearman r = 0.27; p = 0.021). Together, our findings suggest that encapsulation and multinucleated giant cells control M. bovis viability, whereas neutrophils may serve as a cellular biomarker of bacterial proliferation during natural infection. These data integrate host granuloma responses with mycobacterial dissemination and could provide useful immunopathological-based biomarkers of disease severity in natural infection with M. bovis, an important cattle pathogen.     

Effect of massive sympathetic nervous system activation on coronary blood flow and myocardial energy pool

Our previous work indicates that myocardial ischemia could be the mechanism responsible for the left ventricular (LV) dysfunction that frequently develops after massive sympathetic nervous system (SNS) activation. In this study, coronary blood flow (CBF) and myocardial ATP, creatine phosphate, and lactate concentrations were measured after massively activating the SNS of anesthetized rabbits with an intracisternal injection of veratrine. CBF was measured at time 0 (baseline), and at 2, 10, and 20 min after SNS activation in one group, and at 0, 45, 90, and 150 min in a second group. Myocardial ATP, creatine phosphate, and lactate were measured at 0, 2, 10, 20, 90, and 150 min in separate groups of rabbits. SNS activation caused LV dysfunction in approximately 60% of the rabbits. SNS-related increases in CBF kept pace with the increases in myocardial energy demand as determined from the systolic pressure-heart rate product. The subendocardial-to-subepicardial blood flow ratio did not change significantly. Myocardial creatine phosphate concentration was depressed 2 min after SNS activation and remained depressed for at least 20 min. ATP fell continuously and was significantly lower than baseline by 20 min. Tissue lactate concentration was elevated at this time. By 90 min, the concentrations of all three metabolites had recovered. These results indicate that myocardial high-energy phosphate compounds fall after massive SNS activation, but ischemia does not appear to be the underlying mechanism.     

Tumor necrosis factor, cancer and anticancer therapy

Tumor necrosis factor (TNF) is being utilized as an antineoplastic agent for the treatment of patients with locally advanced solid tumors. However, its role in cancer therapy is debated. Although a large body of evidence supports TNF's antineoplastic activity, the cascade of molecular events underlying TNF-mediated tumor regression observed in vivo is still incompletely elucidated. Intriguingly, some pre-clinical findings suggest that TNF may promote cancer development and progression, which has led to propose anti-TNF therapy as a novel approach to malignancies. In the present work, we summarize the molecular biology of TNF with particular regard to its tumor-related properties, and review the experimental and clinical evidence currently available describing the complex and sometime conflicting relationship between this cytokine, cancer and antitumor therapy. Recent insights that might pave the way to further exploitation of the antineoplastic potential of TNF are also discussed.     

Evidence of coprophagy in freshwater zooplankton

1. Vertical transport of nutrients in sedimenting faecal material is greatly reduced by coprophageous organisms. Unfortunately, nearly all work on faecal production, sedimentation and coprophagy has dealt with copepods in marine ecosystems. Here, we report the first evidence of coprophagy in freshwater zooplankton from oligotrophic and eutrophic lakes. We used ¹⁴C‐labelled algae and faecal material to estimate the rates of algal clearance and coprophagy. 2. Measured feeding rates per individual on faecal material were similar (Daphnia pulex, D. rosea, Leptodiaptomus tyrelli) or even higher (D. lumholtzi) than filtering rates on phytoplankton. This finding does not necessarily implicate active selection of faeces over algae because: (i) we did not use the same food concentrations for faeces and algae, and (ii) grazers of slightly different sizes were used in each test. 3. Weight‐specific clearance rates of L. tyrelli and Holopedium gibberum on faecal matter (0.084–0.089 mL μg^?1 h^?1) were higher than in the daphniids (0.026 mL μg^?1 h^?1). 4. The data indicate that coprophagy in freshwater ecosystems is an important mechanism of nutrient recycling, and this process should be taken into account when studying nutrient fluxes within lakes and reservoirs.     

Zooplankton Seasonal Abundance of South AmericanSaline Shallow Lakes

The central provinces of Argentina are characterized by the presence of a high number of shallow lakes, located in endorheic basins, many of which have elevated salinities as well as eutrophic or hypereutrophic condition. The zooplankton of four saline shallow lakes of the province of La Pampa was studied on a monthly basis during a 2‐year period to determine its temporal and spatial variation. The surface of these shallow lakes (<2.5 m depth) varied between 56.8 and 215.9 ha, and some have from 8.4 to 20.8 g · l^–1. The more saline lakes have “clear” water and the less saline lakes “turbid” water. Fishes, Jenynsia multidentata , were present in only two lakes during the last two months of the studied period. The zooplankton was composed of 17 taxa of Rotifera, 5 taxa of Cladocera and 4 taxa of Copepoda. The low diversity and the faunistic composition are characteristic of saline environments. Although the studied lakes share 38% of the species, the faunistic similarity was higher between the two least saline lakes. The lowest diversity was found in the two most saline lakes. All four shallow lakes were characterized by their very high zooplankton density, especially in the least saline lakes (<80000 ind · l^–1). The abundance is significantly correlated with the water transparency but not with salinity. The zooplankton temporal variation was characterized by the alternation of macro‐ and microzooplankton, probably regulated by competition and intrazooplanktonic predation. In each lake, the spatial abundance distribution of the macro‐ and microzooplankton was homogeneous. It was related to the shallow depht of the lakes and their polymictic condition. The Scheffer model on alternative states in shallow lakes acknowledges that it cannot be applied to saline lakes because Daphnia , the main responsible for the clear water state, is not tolerant to high salinity. Our study shows that the most saline lakes, where the halophylic Daphnia menucoensis is abundant, have also the most clear waters. Another difference that we found with regards to the mentioned model is that, in turbid lakes, it could not have had a top‐down control on macrozooplankton exerted by fishes because in these lakes fishes were practically absent. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

P2Y1 receptor-evoked glutamate exocytosis from astrocytes: control by tumor necrosis factor-alpha and prostaglandins

ATP, released by both neurons and glia, is an important mediator of brain intercellular communication. We find that selective activation of purinergic P2Y1 receptors (P2Y1R) in cultured astrocytes triggers glutamate release. By total internal fluorescence reflection imaging of fluorescence-labeled glutamatergic vesicles, we document that such release occurs by regulated exocytosis. The stimulus-secretion coupling mechanism involves Ca2+ release from internal stores and is controlled by additional transductive events mediated by tumor necrosis factor-alpha (TNFalpha) and prostaglandins (PG). P2Y1R activation induces release of both TNFalpha and PGE2 and blocking either one significantly reduces glutamate release. Accordingly, astrocytes from TNFalpha-deficient (TNF(-/-)) or TNF type 1 receptor-deficient (TNFR1(-/-)) mice display altered P2Y1R-dependent Ca2+ signaling and deficient glutamate release. In mixed hippocampal cultures, the P2Y1R-evoked process occurs in astrocytes but not in neurons or microglia. P2Y1R stimulation induces Ca2+ -dependent glutamate release also from acute hippocampal slices. The process in situ displays characteristics resembling those in cultured astrocytes and is distinctly different from synaptic glutamate release evoked by high K+ stimulation as follows: (a) it is sensitive to cyclooxygenase inhibitors; (b) it is deficient in preparations from TNF(-/-) and TNFR1(-/-) mice; and (c) it is inhibited by the exocytosis blocker bafilomycin A1 with a different time course. No glutamate release is evoked by P2Y1R-dependent stimulation of hippocampal synaptosomes. Taken together, our data identify the coupling of purinergic P2Y1R to glutamate exocytosis and its peculiar TNFalpha- and PG-dependent control, and we strongly suggest that this cascade operates selectively in astrocytes. The identified pathway may play physiological roles in glial-glial and glial-neuronal communication.     

Type VI collagen induces cardiac myofibroblast differentiation: implications for postinfarction remodeling

Cardiac fibroblast (CF) proliferation and differentiation into hypersecretory myofibroblasts can lead to excessive extracellular matrix (ECM) production and cardiac fibrosis. In turn, the ECM produced can potentially activate CFs via distinct feedback mechanisms. To assess how specific ECM components influence CF activation, isolated CFs were plated on specific collagen substrates (type I, III, and VI collagens) before functional assays were carried out. The type VI collagen substrate potently induced myofibroblast differentiation but had little effect on CF proliferation. Conversely, the type I and III collagen substrates did not affect differentiation but caused significant induction of proliferation (type I, 240.7 +/- 10.3%, and type III, 271.7 +/- 21.8% of basal). Type I collagen activated ERK1/2, whereas type III collagen did not. Treatment of CFs with angiotensin II, a potent mitogen of CFs, enhanced the growth observed on types I and III collagen but not on the type VI collagen substrate. Using an in vivo model of myocardial infarction (MI), we measured changes in type VI collagen expression and myofibroblast differentiation after post-MI remodeling. Concurrent elevations in type VI collagen and myofibroblast content were evident in the infarcted myocardium 20-wk post-MI. Overall, types I and III collagen stimulate CF proliferation, whereas type VI collagen plays a potentially novel role in cardiac remodeling through facilitation of myofibroblast differentiation.