Comparison between Urticaceae (Parietaria) pollen count and hay fever symptoms: assessment of a «threshold-value»

Summary A comparison betweenParietaria pollen count and allergic symptoms of rhino-conjunctivitis in the early season was used in utilized to determine a «threshold-value» for this pollen. Clinical data were obtained from diary-cards of 34 allergic patients and pollen data from a volumetric sampling, carried out by means of a Hirst-Burkard pollen-trap. A significant correlation (r=0.98) was found between pollen count and symptom scores. Mild symptoms were registered with concentrations above 10–15 pollens/m³. Severe symptoms occurred when pollen count exceeded 80/m³/24 h., and over 90% of patients recorded symptoms. The importance of the late reactions and of the total allergenic airborne content are emphasized.     

The first structure of a bacterial class B Acid phosphatase reveals further structural heterogeneity among phosphatases of the haloacid dehalogenase fold

AphA is a periplasmic acid phosphatase of Escherichia coli belonging to class B bacterial phosphatases, which is part of the DDDD superfamily of phosphohydrolases. The crystal structure of AphA has been determined at 2.2A and its resolution extended to 1.7A on an AuCl(3) derivative. This represents the first crystal structure of a class B bacterial phosphatase. Despite the lack of sequence homology, the AphA structure reveals a haloacid dehalogenase-like fold. This finding suggests that this fold could be conserved among members of the DDDD superfamily of phosphohydrolases. The active enzyme is a homotetramer built by using an extended N-terminal arm intertwining the four monomers. The active site of the native enzyme, as prepared, hosts a magnesium ion, which can be replaced by other metal ions. The structure explains the non-specific behaviour of AphA towards substrates, while a structure-based alignment with other phosphatases provides clues about the catalytic mechanism.     

Spatiotemporal image correlation spectroscopy (STICS) theory, verification, and application to protein velocity mapping in living CHO cells

We introduce a new extension of image correlation spectroscopy (ICS) and image cross-correlation spectroscopy (ICCS) that relies on complete analysis of both the temporal and spatial correlation lags for intensity fluctuations from a laser-scanning microscopy image series. This new approach allows measurement of both diffusion coefficients and velocity vectors (magnitude and direction) for fluorescently labeled membrane proteins in living cells through monitoring of the time evolution of the full space-time correlation function. By using filtering in Fourier space to remove frequencies associated with immobile components, we are able to measure the protein transport even in the presence of a large fraction (>90%) of immobile species. We present the background theory, computer simulations, and analysis of measurements on fluorescent microspheres to demonstrate proof of principle, capabilities, and limitations of the method. We demonstrate mapping of flow vectors for mixed samples containing fluorescent microspheres with different emission wavelengths using space time image cross-correlation. We also present results from two-photon laser-scanning microscopy studies of alpha-actinin/enhanced green fluorescent protein fusion constructs at the basal membrane of living CHO cells. Using space-time image correlation spectroscopy (STICS), we are able to measure protein fluxes with magnitudes of mum/min from retracting lamellar regions and protrusions for adherent cells. We also demonstrate the measurement of correlated directed flows (magnitudes of mum/min) and diffusion of interacting alpha5 integrin/enhanced cyan fluorescent protein and alpha-actinin/enhanced yellow fluorescent protein within living CHO cells. The STICS method permits us to generate complete transport maps of proteins within subregions of the basal membrane even if the protein concentration is too high to perform single particle tracking measurements.     

A mathematical description of blood spiral flow in vessels: application to a numerical study of flow in arterial bending

Local arterial haemodynamics has been associated with the pathophysiology of several cardiovascular diseases. The stable spiral blood-flows that were observed in vivo in several vessels, may play a dual role in vascular haemodynamics, beneficial since it induces stability, reducing turbulence in the arterial tree, and accounts for normal organ perfusion, but detrimental in view of the imparted tangential velocities that are involved in plaque formation and development. Being a spiral flow considered representative of the local blood dynamics in certain vessels, a method is proposed to quantify the spiral structure of blood flow. The proposed function, computed along a cluster of particle trajectories, has been tested for the quantitative determination of the spiral blood flow in a three-dimensional, s-shaped femoral artery numerical model in which three degrees of stenosis were simulated in a site prone to atherosclerotic development. Our results confirm the efficacy of the Lagrangian analysis as a tool for vascular blood dynamics investigation. The proposed method quantified spiral motion, and revealed the progression in the degree of stenosis, in the presented case study. In the future, it could be used as a synthetic tool to approach specific clinical complications.     

Chaos and population control of insect outbreaks

We used small perturbations in adult numbers to control large fluctuations in the chaotic demographic dynamics of laboratory populations of the flour beetle Tribolium castaneum . A nonlinear mathematical model was used to identify a sensitive region of phase space where the addition of a few adult insects would result in a dampening of the life stage fluctuations. Three experimental treatments were applied: one in which perturbations were made whenever the populations were inside the sensitive region ("in-box treatment"), another where perturbations were made whenever the populations were outside the sensitive region ("out-box treatment"), and an unperturbed control. The in-box treatment caused a stabilization of insect densities at numbers well below the peak values exhibited by the out-box and control populations. This study demonstrates how small perturbations can be used to influence the chaotic dynamics of an ecological system.     

Mitochondria are involved in the neurogenic neuroprotection conferred by stimulation of cerebellar fastigial nucleus

Activation of neural pathways originating in the cerebellar fastigial nucleus (FN) protects the brain from the deleterious effects of cerebral ischemia and excitotoxicity, a phenomenon termed central neurogenic neuroprotection. The neuroprotection is, in part, mediated by suppression of apoptosis. We sought to determine whether FN stimulation exerts its anti-apoptotic effect through mitochondrial mechanisms. Mitochondria were isolated from the cerebral cortex of rats in which the FN was stimulated for 1 h (100 microA; 1 s on/1 s off), 72 h earlier. Stimulation of the dentate nucleus (DN), a brain region that does not confer neuroprotection, served as control. Mitochondria isolated from FN-stimulated rats exhibited a marked increase in their ability to sequester Ca2+ and an increased resistance to Ca2+-induced membrane depolarization and depression in respiration. FN stimulation also leads to reduction in the release in cytochrome c, induced either by Ca2+ or the mitochondrial toxin mastoparan. Furthermore, in brain slices, FN stimulation reduced the staurosporine-induced insertion of the pro-apoptotic protein Bax into the mitochondria, a critical step in the mitochondrial mechanisms of apoptosis. Collectively, these results provide evidence that FN stimulation protects the mitochondria from dysfunction induced by Ca2+ loading, and inhibits mitochondrial pathways initiating apoptosis. These mitochondrial mechanisms are likely to play a role in the neuroprotection exerted by FN stimulation.     

Comparison of serum levels of seven cytokines in premature newborns undergoing different ventilatory procedures: high frequency oscillatory ventilation or synchronized intermittent mandatory ventilation

OBJECTIVE: The severity of pulmonary dysfunction and subsequent development of chronic lung disease (CLD) in preterm neonates depends on several factors, among them oxygen administration. The aim of this report is to compare the effects of high-frequency, oscillatory ventilation (HFOV) versus synchronized, intermittent, mandatory ventilation (sIMV) on serum cytokine levels (IL-6, IL-8, IL-10, MCP-1, PDGF-BB, VEGF and TGF-beta1) and ventilator indices during the first week of life. Moreover, CLD development and several other outcomes were compared between the two groups. DESIGN: Randomized clinical trial. SETTING: Third level NICU. PATIENTS: 40 preterm neonates with a gestational age between 24 and 29 weeks were randomly (20 per group) assigned to one of the two, above-mentioned ventilation strategies within 30 minutes of birth. MEASUREMENTS AND RESULTS: At 1, 3 and 5 days, neonates were monitored by means of ventilator indices and levels of seven pro-inflammatory or anti-inflammatory (pro-fibrotic) cytokines in serum. No clinical or biochemical differences were observed at baseline. The neonates assigned to HFOV benefited from early and sustained improvement in gas exchange, with earlier extubation and lower incidence of CLD, as compared to the neonates assigned to sIMV treatment, and showed a significant reduction of serum IL-6, IL-8 and IL-10 over time only when the HFOV treatment was administered. In addition, at days 3 and 5, the IL-6 levels were significantly lower in the HFOV group as compared to sIMV patients. CONCLUSIONS: The results of this randomized clinical trial support the hypothesis that early use of HFOV, combined with an optimum volume strategy, has a beneficial effect, reducing serum levels of pro-inflammatory cytokines and consequently the acute phase leading to lung injury.     

Docking studies on PARP-1 inhibitors: insights into the role of a binding pocket water molecule

The binding mode of a series of competitive PARP-1 inhibitors was investigated employing a molecular docking approach by using Autodock 3.0. A particular attention was given to the role played by a water molecule present in some but not all the so far available crystal structures of the catalytic domain of PARP-1. Good correlation between calculated binding energies and experimental inhibitory activities was obtained either by including (r2=0.87) or not (r2=0.84) the structural water molecule. Closer inspection of our results suggested that this water molecule should be considered part of the hydration shell of polar inhibitors and not as a structural water.