Analysis of pulmonary arterial pressure profile after occlusion of pulsatile blood flow

In isolated canine lung lobes perfused with a pulsatile pump, arterial occlusions were performed and the postocclusion arterial pressure profiles were analyzed to estimate the pulmonary capillary pressure. A solenoid valve interposed between the pump and the lobar artery was used to perform arterial occlusions at several instants equally distributed within a pressure cycle. Double occlusions were also accomplished by simultaneously activating the solenoid valve and clamping the venous outflow of the lung lobe. To analyze an arterial occlusion pressure profile, we computed the best monoexponential fit of the pressure decay over a short period of time after the occlusion maneuvers. Two estimates of the capillary pressure were derived from this analysis: 1) the extrapolation of the exponential fit to the instant of occlusion, and 2) the point at which the recorded pressure decay curve merges with the exponential fit. The pressures thus determined were compared with the double occlusion pressure that provided an independent estimate of the pulmonary capillary pressure. Our results show that, under a wide range of conditions, the estimates of the capillary pressure obtained from the arterial occlusion data are nearly equal to the double occlusion pressures. Additionally, we estimated the capillary pressure variations within a pressure cycle by examining the occlusion pressures sampled at different instants of the cycle. The pulsatility of the pulmonary microvascular pressure varied with the pump frequency as well as the state of arterial and venous vasoaction. These variations are consistent with the representation of the lung vasculature as a low-pass filter.     

Nox-generated ROS modulate glucose uptake in a leukaemic cell line

The discovery of superoxide-generating enzymes homologues of phagocytic NAD(P)H oxidase, the Nox family, has led to the concept that reactive oxygen species (ROS) are 'intentionally' generated with biological functions in various cell types. In this study, by treating an acute leukaemic cell line with different antioxidants, ROS generation was shown to be crucially involved in the modulation of glucose transport (mediated by Glut1), which is frequently up-regulated in cancer cells. Then, this study tried to elucidate ROS source(s) and mechanisms by which ROS are involved in Glut1 activity regulation. Results prove that Nox2 and Nox4 are the candidates and that phosphorylation processes are important in the regulation of glucose uptake on which cancer cells rely. On the whole, data suggest that both Glut1 and Nox homologues may be considered new potential targets in the treatment of leukaemia.     

Petrobactin is produced by both pathogenic and non-pathogenic isolates of the Bacillus cereus group of bacteria

Petrobactin is the primary siderophore synthesized by Bacillus anthracis str Sterne and is required for virulence of this organism in a mouse model. The siderophore's biosynthetic machinery was recently defined and gene homologues of this operon exist in several other Bacillus strains known to be mammalian pathogens, but are absent in several known to be harmless such as B. subtilis and B. lichenformis. Thus, a common hypothesis regarding siderophore production in Bacillus species is that petrobactin production is exclusive to pathogenic isolates. In order to test this hypothesis, siderophores produced by 106 strains of an in-house library of the Bacillus cereus sensu lato group were isolated and identified using a MALDI-TOF-MS assay. Strains were selected from a previously defined phylogenetic tree of this group in order to include both known pathogens and innocuous strains. Petrobactin is produced by pathogenic strains and innocuous isolates alike, and thus is not itself indicative of virulence.     

Scn3b knockout mice exhibit abnormal ventricular electrophysiological properties

We report for the first time abnormalities in cardiac ventricular electrophysiology in a genetically modified murine model lacking the Scn3b gene (Scn3b^−/−). Scn3b^−/− mice were created by homologous recombination in embryonic stem (ES) cells. RT-PCR analysis confirmed that Scn3b mRNA was expressed in the ventricles of wild-type (WT) hearts but was absent in the Scn3b^−/− hearts. These hearts also showed increased expression levels of Scn1b mRNA in both ventricles and Scn5a mRNA in the right ventricles compared to findings in WT hearts. Scn1b and Scn5a mRNA was expressed at higher levels in the left than in the right ventricles of both Scn3b^−/− and WT hearts. Bipolar electrogram and monophasic action potential recordings from the ventricles of Langendorff-perfused Scn3b^−/− hearts demonstrated significantly shorter ventricular effective refractory periods (VERPs), larger ratios of electrogram duration obtained at the shortest and longest S₁–S₂ intervals, and ventricular tachycardias (VTs) induced by programmed electrical stimulation. Such arrhythmogenesis took the form of either monomorphic or polymorphic VT. Despite shorter action potential durations (APDs) in both the endocardium and epicardium, Scn3b^−/− hearts showed ΔAPD₉₀ values that remained similar to those shown in WT hearts. The whole-cell patch-clamp technique applied to ventricular myocytes isolated from Scn3b^−/− hearts demonstrated reduced peak Na⁺ current densities and inactivation curves that were shifted in the negative direction, relative to those shown in WT myocytes. Together, these findings associate the lack of the Scn3b gene with arrhythmic tendencies in intact perfused hearts and electrophysiological features similar to those in Scn5a^+/− hearts.     

A large number of the human microRNAs target lentiviruses, retroviruses, and endogenous retroviruses

Retroelements (including transposons, retrotransposons, retroviruses, and lentiviruses) make up a significant portion of eukaryotic genomes. Given their ability to mutate genes these mobile elements always present a threat to the integrity of the host genomes. Recent studies have revealed complex molecular mechanisms that silence the mutagenic ability of these RE as well strategically express the pieces of the incorporated RE that are utilized to silence human endogenous retroviruses (HERVs) or invading exogenous retroviruses (IERV). We have hypothesized that small endogenous RNA originally evolved to quell “foreign” IERV-genes and subsequently emerged into elaborate silencing systems that include RNA interference, miRNA-based gene regulation and other gene silencing mechanisms. Here, we present evidence that the replication of complex RE are most likely silenced or regulated by homologous miRNA that are found as a part of the cellular repertoire. We analyzed Homo sapiens miRNAs for possible target genetic sequences in selected HERVs and IERV found in humans and other large primates. We identified several miRNAs that have >80% sequence homology with human HERVs; -L, -W, and -K, and IERV like SIVcpz, HTLV-1, and HTLV-2. We found an inverse correlation between the numbers and relative degree of homology of miRNAs to the relative replication capacity of a specific RE. Therefore, larger numbers of miRNAs with greater degree of homology are found against the least active RE and the least numbers of miRNAs with smaller degree of homology are found against the most active RE (i.e. HERV-K). Implications of these observations in RE disease and therapy are discussed.     

Regeneration potential of the submandibular gland after transplantation: An immunohistological and scintigraphic study

The aim of this study was to evaluate the regeneration potential of the submandibular gland and the associated morphological and functional impairment after a transient warm ischemia as a result of submandibular gland transfer in humans. 42 rabbits were used for the study. After 1.5 h of transient ischemia, submandibular glands were studied histologically in the following 14 days, one month, two and four months in 32 rabbits. Additionally, the glands were functionally evaluated after one and four months in eleven of these rabbits. Ten rabbits were used to establish the scintigraphic method. In the evaluation of the results unpublished data derived from a previous study by our group has also been considered. The Ki-67 index showed a significant increase of the proliferating rate reaching a peak after 14 days (p = 0.006) and still evident after four months involving mostly the terminal ductal system. There was only a slight increase in the tracer uptake of ischemic glands after one month and a more pronounced one after four months.It was concluded that initial loss of acinar cells by 1.5 h of ischemia released a regeneration process which stabilised after 14 days but continued throughout the whole time of the study. These results correlate with clinical observation in patients with submandibular gland transfer for the treatment of keratoconjunctivitis sicca and explain the alternating secretory function observed within the four months following surgery. Scintigraphy has been found to play a limited role in the evaluation of the regeneration process.     

Toxicological Study and Oxidative Stress Evaluation for Safety Assessment of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Vip3Aa16 Toxin in Adult Mice

Vegetative insecticidal proteins were produced by some Bacillus thuringiensis strains and were successfully used in biological control against different agricultural pests such as Lepidoptera. To assess the safety of Vip3Aa16 toxins in mammalian organisms, we evaluated their toxicity using histological, hematological, and oxidative stress parameters on albino Swiss mice. The animals were orally treated with 2500, 5000, 7500 milligrams (mg) of the toxin/kilogram (kg) of body weight for 14 days. Then samples of blood, kidney and hepatic tissues were collected at the end of the treatment. Hematological parameters were monitored by RBC, WBC, hemoglobin, hematocrit, MCV, MCH, and MCHC. Liver and kidney MDA, SOD, vitamin C and H₂O₂ were analyzed to assess oxidative damage. Hepatotoxicity was monitored by analysis of the plasma enzymes ALT and AST and bilirubin levels. Renal toxicity was tested by urea, uric acid and creatinine evaluation. The histopathology of kidney and liver tissues was also investigated. The results of the toxicological study revealed that the Vip3AaA16 has no lethal effect since no mortality was observed at any dose. Moreover, body weight, hematological, histological, biochemical and oxidative findings showed no significant differences between treated and control groups. All these findings confirmed that this toxin is highly safe and doesn’t represent any risk on animal health and subsequently, Vip3Aa16 toxin can be safely used in biological programs to control Lepidopteran pests attacking crops around the world.