Commonly used numbers of microspheres affect cardiac vascular resistance

The main goal of the study was to examine how the microsphere technique affects the hemodynamics and mitochondrial energy status of the Langendorff-perfused rat heart. The hearts were perfused at a constant flow with Tyrode solution. NADH videofluorometry of the surface of the left ventricle was used to record the mitochondrial energy status as indication of regional ischemia. The effects of seven successive (separated by 10 min) injections of 0.1 ml of saline or (0.05% Tween 20; polysorbate 20, Sigma-Aldrich, St. Louis, MO, U.S.A.) or (0.05% Tween 20 + microspheres) were studied. The number of microspheres per injection were: #1 (2,500), #2 (5,000), #3 (10,000), #4 (20,000), #5 (40,000), #6 (40000), and #7 (80000). The anti-aggregation agent Tween always caused a biphasic response in perfusion pressure. Compared with the Tween effect, the injection of microspheres caused an initial change (mm Hg) in perfusion pressure of #1 (-10), #2 (NS), #3 (NS), #4 (+7.5), #5 (+12.3), #6 (+14.4), #7 (+18.3), and a delayed change (10 min after injection) of #1 (-22.2), #2 (-6.0), #3 (-4.1), #4 (-4.5), #5 (NS), #6 (NS), and #7 (+5.9). The microspheres caused a significant delayed increase in NADH only for injection #6 and #7. Similar results were found for different durations of the input function or when hearts were perfused at constant perfusion pressure. In hearts without flow reserve (10 microM adenosine), Tween injections were without effect, whereas three successive injections of 60,000 microspheres each only caused increases in perfusion pressure and NADH. The data demonstrate that in hearts with flow reserve present, even very low numbers of microspheres (2,000/g heart) cause large decreases in perfusion pressure without obvious signs of ischemia. When flow reserve was exhausted by either microsphere loading or adenosine addition, microspheres only caused increases in perfusion pressure and resulted in detectable ischemia (NADH). It is concluded that microspheres affect the vascular resistance of the heart and that these effects are flow reserve dependent.     

Protective effects of fungal (1-->3)-beta-D-glucan derivatives against oxidative DNA lesions in V79 hamster lung cells

We have investigated the protein turnover modulations involved in the hypertrophic muscle phenotype of c-ski overexpressing transgenic mice. In these animals, the body weight is increased and all the muscles examined show a definite hypertrophy. The protein degradation rate is significantly reduced in the fast twitch muscles of c-ski transgenic animals with respect to controls; in contrast, there are no detectable differences in the synthesis rates. The down-regulation of protein breakdown is paralleled by decreased expression of genes belonging to the lysosomal as well as to the ATP-ubiquitin-dependent proteolytic pathways.     

Size effect of intratracheally instilled particles on pulmonary inflammation and vascular thrombosis

Particulate air pollution is associated with cardiorespiratory effects and ultrafine particles (UFPs, diameter < 100 nm) are believed to play an important role. We studied the acute (1 h) effect of intratracheally instilled unmodified (60 nm), negatively charged carboxylate-modified (60 nm), or positively charged amine-modified (60 or 400 nm) polystyrene particles on bronchoalveolar lavage (BAL) indices and on peripheral thrombosis in hamster. The latter was assessed by measuring the extent of photochemically induced thrombosis in a femoral vein via transillumination. Unmodified and negative UFPs did not modify thrombosis and BAL indices. Positive UFPs increased thrombosis at 500 microg per animal (+ 341 +/- 96%) and at 50 microg per animal (+ 533 +/- 122%), but not at 5 microg per animal. Neutrophils, lactate dehydrogenase, and histamine were increased in BAL at all these doses but protein concentration was increased only at 500 microg per animal. Positive 400-nm particles (500 microg per animal) did not affect thrombosis, although they led to a neutrophil influx and an increase in BAL proteins and histamine. Using the Platelet Function Analyser (PFA-100), the platelets of hamsters were activated by the in vitro addition of positive UFPs and 400-nm particles to blood. We conclude that intratracheally administered positive ultrafine and 400-nm particles induce pulmonary inflammation within 1 h. Positive UFPs, but not the 400-nm particles enhance thrombosis. Hence, particle-induced lung inflammation and thrombogenesis can be partially uncoupled.     

Nonsyndromic mental retardation and cryptogenic epilepsy in women with doublecortin gene mutations

DCX mutations cause mental retardation in male subjects with lissencephalypachygyria and in female subjects with subcortical band heterotopia (SBH). We observed four families in which carrier women had normal brain magnetic resonance imaging (MRI) and mild mental retardation, with or without epilepsy. Affected male subjects had SBH or pachygyria-SBH. In two families, the phenotype was mild in both genders. In the first family, we found a tyr138his mutation that is predicted to result in abnormal folding in the small hinge region. In the second family, we found an arg178cys mutation at the initial portion of R2, in the putative beta-sheet structure. Carrier female subjects with normal MRI showed no somatic mosaicism or altered X-inactivation in lymphocytes, suggesting a correlation between mild mutations and phenotypes. In the two other families, with severely affected boys, we found arg76ser and arg56gly mutations within the R1 region that are predicted to affect DCX folding, severely modifying its activity. Both carrier mothers showed skewed X-inactivation, possibly explaining their mild phenotypes. Missense DCX mutations may manifest as non-syndromic mental retardation with cryptogenic epilepsy in female subjects and SBH in boys. Mutation analysis in mothers of affected children is mandatory, even when brain MRI is normal.     

Towards an understanding of biological role of colostrinin peptides

The aim of this study was to elucidate the structure and possible function of colostrinin, also known as a proline rich polypeptide (PRP). The molecular weight of colostrinin was originally determined by gel filtration to be 17,200 daltons. In the presence of guanidinum chloride, however, the molecular weight was found to be about 6,000 daltons. Further studies utilizing high-performance liquid chromatography (HPLC) and mass spectroscopy revealed that colostrinin is a complex consisting of many low molecular-weight polypeptides. A total of 32 peptides were isolated from the original colostrinin preparation by HPLC and subjected to the N-terminal sequence analysis. The results of sequence analysis revealed significant homology of the peptides to three protein precursors: annexin, β-casein, and a hypothetical β-casein homolog. In addition, the sequence of several peptides showed no significant homology to any specific protein in the current GenBank database. The synthetic peptides of various lengths representing the N-terminal sequence of the colostrinin peptides were made to study some biological effects. Here we report that colostrinin and some of its component peptides are potent inducers of leukocyte proliferation and of certain cytokines. Also, a series of monospecific antibodies were produced in rabbits against the synthetic peptides. The antibodies were used to study the kinetic of antigen reduction in colostrum and mature milk following lambing. A threefold decrease was common for most antigens studied over the period of 72 h. Based on the results of these studies we postulate that colostrinin represents a diverse group of peptides produced in the mammary gland of mammals for the development of the optimal physiologic responses in offspring. Also, it is hoped that the beneficial use of colostrinin in Alzheimer’s Disease (AD), recently reported elsewhere, will revive interest in its clinical application for treatment and/or prophylaxis of many age-related disorders.     

Intensive care delirium monitoring and standardised treatment: A complete survey of Dutch Intensive Care Units

OBJECTIVE: Delirium is a frequent and serious problem in the Intensive Care Unit (ICU). Several international guidelines recommend daily monitoring for ICU-delirium. The purpose of this article is to give an up-to-date overview of the current status of monitoring and treatment of ICU-delirium in the Netherlands. DESIGN: Nation-wide, telephone-based questionnaire survey. PARTICIPANTS: Head nurse of all ICUs and a random sample of intensivists. RESULTS: Only 14% (n=14) of all Dutch ICUs (n=103) monitored for ICU-delirium. Of these, only half (7%) used a tool that is validated in ICU patients. In 31% of Dutch ICUs, a protocol was used to treat ICU-delirium. Responses were obtained from 100% of ICUs. CONCLUSION: Despite an international guideline, not more than 7% of ICUs in our study routinely evaluated the presence of delirium with a validated instrument. Fewer than one-third of Dutch ICUs use a protocol to treat ICU-delirium.     

Activation of Coagulation Factor V by a Platelet Protease

Factor V must be converted to Factor V_a in order to bind to a high affinity platelet surface site and participate in prothrombin activation. Osterud et al. (10) presented data that suggested that human platelets contain an activated form of Factor V and a Factor V activator. We find that the Factor V released when platelets are disrupted by freezing and thawing or sonication is activated 3- to 10-fold by thrombin as determined by coagulation assay and is therefore stored as the relatively inactive procofactor rather than in the active form Factor V_a.     

Left atrial volume determination by echocardiography

To validate echocardiographic left atrial volume measurements, 25 patients with mitral stenosis were studied before and after mitral balloon valvuloplasty. Seven normals served as controls. The modified Simpson's rule was used for echocardiographic and angiographic left atrial volume determination from two orthogonal planes. Left atrial antero-posterior diameter was measured from parasternal long axis view and supero-inferior and medio-lateral diameters from apical four-chamber view. Transthoracic echocardiographic left atrial volume correlated well, but systematically underestimated angiographic left atrial volume (y=0.4x+27, r=0.92). Monoplane transesophageal echocardiography did not improve correlation, nor the underestimation. Out of the several left atrial diameters, antero-posterior dimension showed the closest correlation with angiographic volume (r=0.91), which persisted after exclusion of patients with atria >400 ml (r=0.84). Futhermore, relative changes of antero-posterior diameter after mitral valvuloplasty were closely related to the relative changes observed in left atrial volume (r=0.82). Our results suggest that, in spite of a consistent underestimation, bidimensional, transthoracic echocardiographic and angiographic left atrial assessment correlate closely. Moreover, it is suggested that the mere antero-posterior diameter from transthoracic two-dimensional image is sufficient in clinical practice for routine follow-op of left atrial volume.