Primary Aldosteronism and Resistant Hypertension: A Pathophysiological Insight

Primary aldosteronism (PA) is a pathological condition characterized by an excessive aldosterone secretion; once thought to be rare, PA is now recognized as the most common cause of secondary hypertension. Its prevalence increases with the severity of hypertension, reaching up to 29.1% in patients with resistant hypertension (RH). Both PA and RH are “high-risk phenotypes”, associated with increased cardiovascular morbidity and mortality compared to non-PA and non-RH patients. Aldosterone excess, as occurs in PA, can contribute to the development of a RH phenotype through several mechanisms. First, inappropriate aldosterone levels with respect to the hydro-electrolytic status of the individual can cause salt retention and volume expansion by inducing sodium and water reabsorption in the kidney. Moreover, a growing body of evidence has highlighted the detrimental consequences of “non-classical” effects of aldosterone in several target tissues. Aldosterone-induced vascular remodeling, sympathetic overactivity, insulin resistance, and adipose tissue dysfunction can further contribute to the worsening of arterial hypertension and to the development of drug-resistance. In addition, the pro-oxidative, pro-fibrotic, and pro-inflammatory effects of aldosterone may aggravate end-organ damage, thereby perpetuating a vicious cycle that eventually leads to a more severe hypertensive phenotype. Finally, neither the pathophysiological mechanisms mediating aldosterone-driven blood pressure rise, nor those mediating aldosterone-driven end-organ damage, are specifically blocked by standard first-line anti-hypertensive drugs, which might further account for the drug-resistant phenotype that frequently characterizes PA patients.     

Lysophosphatidylinositol Induced Morphological Changes and Stress Fiber Formation through the GPR55-RhoA-ROCK Pathway

We previously reported that lysophosphatidylinositol (LPI) functions as an endogenous agonist of GPR55, a novel cannabinoid receptor. However, the physiological roles of LPI-GPR55 have not yet been elucidated in detail. In the present study, we found that LPI induced morphological changes in GPR55-expressing HEK293 cells. LPI induced the cell rounding of GPR55-expressing HEK293 cells but not of empty-vector-transfected cells. LPI also induced the activation of small GTP-binding protein RhoA and increased stress fiber formation in GPR55-expressing HEK293 cells. The inhibition of RhoA and Rho kinase ROCK by the C3 exoenzyme and the ROCK inhibitor reduced LPI-induced cell rounding and stress fiber formation. These results clearly indicated that the LPI-induced morphological changes and the assembly of the cytoskeletons were mediated through the GPR55-RhoA-ROCK pathway.     

Prolidase activity in chronic wound and blister fluids

We investigated prolidase activity in samples derived from wound fluid as well as blister fluid. Prolidase activity was elevated in fluid samples collected from wounds over the levels in sera collected from patients with chronic wounds (P < 0.05). Prolidase activity was also present in samples taken from blister diseases. However, prolidase activity in blister fluid was not higher than that in sera collected from patients with blister diseases. Our results indicate that prolidase may play a role in wound healing.     

Knowledge-based 3D analysis from 2D medical images

An anatomical knowledge-based system for image analysis that interprets CT/MR (computed tomography/magnetic resonance) images of the human chest cavity is reported. The approach utilizes a low-level image analysis system with the ability to analyze the data in bottom-up (or data-driven) and top-down (or model-driven) modes to improve the high-level recognition process. Several image segmentation algorithms, including K-means clustering, pyramid-based region extraction, and rule-based merging, are used for obtaining the segmented regions. To obtain a reasonable number of well-segmented regions that have a good correlation with the anatomy, a priori knowledge in the form of masks is used to guide the segmentation process. Segmentation of the brain is also considered.     

Contact Sex Pheromone Components of the Seed Beetle, <Emphasis Type="BoldItalic">Callosobruchus analis</Emphasis> (F.)

Callosobruchus analis (Coleoptera: Chrysomelidae: Bruchinae), found throughout tropical Asia and Africa, is a pest of stored legumes. Previous work has shown that females of this species produce a contact sex pheromone that elicits copulatory behavior in males. Comparisons of copulatory activity between any two of four congeneric species suggest that the contact sex pheromones are species specific. In laboratory bioassays, male C. analis exhibited copulatory behavior to a female dummy to which a crude extract of virgin females was applied. The extract had been collected by a filter paper method and was purified by acid–base partition and chromatographic techniques. Gas chromatography-mass spectrometry (GC-MS) analyses of active fractions revealed that the active compounds were 2,6-dimethyloctane-1,8-dioic acid (1) and callosobruchusic acid, (E)-3,7-dimethyl-2-octene-1,8-dioic acid (2), previously identified as contact sex pheromones of Callosobruchus maculatus (F.) and C. chinensis (L.), respectively. The stereoisomeric and chemical compositions were determined by the 2D-HPLC-Ohrui-Akasaka method as (2S,6R)-1:(S)-2 = 1.8:1, which meant that both compounds in C. analis were stereochemically pure, unlike the case of C. maculatus and C. chinensis. An examination of the influence of synthetic pheromone compounds on male copulatory activity revealed that (2S,6R)-1 is the main component, and that (S)-2 has an additive effect. In the examination of the stereochemistry-activity relationship, no copulatory behavior was elicited by (2R,6S)-1, and, furthermore, the enantiomer significantly masked the pheromonal activity of (2S,6R)-1. Glass rod dummy assays also suggested the presence of synergists. These results could elucidate the specificity of mate recognition in C. analis.     

Manufacturing process and thermal characterization of a fast temperature switching microdevice for real-time biological experiments

Rapid switching and measurement of the temperature in a small volume of liquid are of paramount importance for the real-time observation of biomolecular events such as protein folding–unfolding or enzymatic reactions. We present a microthermodevice for high-speed biological measurements that combines a heater for fast temperature shifting (millisecond range) and a Thin Film Thermocouple (TFTC) for accurate temperature measurement during microscopic observation. Device manufacturing as well as its thermal characterization are shown. The device is successfully employed to measure the fluorescent quenching of double-stranded DNA coupled with fluorescent intercalator on millisecond time scale, thus demonstrating its capacity for innovative biological experiments.     

Availability-based simulation and optimization modeling framework for open-pit mine truck allocation under dynamic constraints

We present a novel system productivity simulation and optimization modeling framework in which equipment availability is a variable in the expected productivity function of the system. The framework is used for allocating trucks by route according to their operating performances in a truck–shovel system of an open-pit mine, so as to maximize the overall productivity of the fleet. We implement the framework in an originally designed and specifically developed simulator–optimizer software tool. We make an application on a real open-pit mine case study taking into account the stochasticity of the equipment behavior and environment. The total system production values obtained with and without considering the equipment reliability, availability and maintainability (RAM) characteristics are compared. We show that by taking into account the truck and shovel RAM aspects, we can maximize the total production of the system and obtain specific information on the production availability and productivity of its components.     

2,3-Dihydrohomofarnesal: Female Sex Attractant Pheromone Component of Callosobruchus rhodesianus (Pic)

Callosobruchus rhodesianus (Pic) (Coleoptera: Chrysomelidae: Bruchinae) is a pest of stored legumes through the Afro-tropical region. In laboratory bioassays, males of C. rhodesianus were attracted to volatiles collected from virgin females. Collections were purified by various chromatographic techniques, and the biologically active component isolated using gas chromatographic-electroantennographic detection analysis. Gas chromatography-mass spectrometry and NMR analyses suggested that the active compound was 2,3-dihydrohomofarnesal, i.e., 7-ethyl-3,11-dimethyl-6,10-dodecadienal. The structure was confirmed by non-stereoselective and enantioselective total synthesis. Using chiral gas chromatography, the absolute configuration of the natural compound was confirmed as (3S,6E)-7-ethyl-3,11-dimethyl-6,10-dodecadienal. Y-tube olfactomter assays showed that only the (S)-enantiomer attracted males of C. rhodesianus. The (R)-enantiomer and racemate did not attract males, suggesting that the (R)-enantiomer inhibits the activity of the natural compound. In combination with previous reports about sex attractant pheromones of congeners, we suggest that a saltational shift of the pheromone structure arose within the genus Callosobruchus.     

Reaction time and EMG measurement applied to human control modeling

The final goal of this research is to analyze the human visual and force sensory feedback integration related to a manipulation task and build a control model of a human operator. In this primary work the reaction time and the EMG was measured to determine the time lag constant of each subject. The onset time, EMG-RT and the EMD delay was measured and compared to the time lag estimated from step response identification. Although a small discrepancy was found between the directly measured time lag and the estimated one, the system identification method showed a prominent and methodological approach to model the human control system.