Surfactants: physicochemical interactions with biological macromolecules.

Macromolecules are essential cellular components in biological systems responsible for performing a large number of functions that are necessary for growth and perseverance of living organisms. Proteins, lipids and carbohydrates are three major classes of biological macromolecules. To predict the structure, function, and behaviour of any cluster of macromolecules, it is necessary to understand the interaction between them and other components through basic principles of chemistry and physics. An important number of macromolecules are present in mixtures with surfactants, where a combination of hydrophobic and electrostatic interactions is responsible for the specific properties of any solution. It has been demonstrated that surfactants can help the formation of helices in some proteins thereby promoting protein structure formation. On the other hand, there is extensive research towards the use of surfactants to solubilize drugs and pharmaceuticals; therefore, it is evident that the interaction between surfactants with macromolecules is important for many applications which includes environmental processes and the pharmaceutical industry. In this review, we describe the properties of different types of surfactants that are relevant for their physicochemical interactions with biological macromolecules, from macromolecules-surfactant complexes to hydrophobic and electrostatic interactions.

The effect of sophorolipids against microbial biofilms on medical-grade silicone.

Recent medical strategies rely on the search for effective antimicrobials as surface coatings to prevent and treat infections in humans and animals. Biosurfactants have recently been shown to have properties as antiadhesive and antibiofilm agents. Sophorolipids in particular are biosurfactant molecules known to act as therapeutic agents. This study aimed to evaluate antimicrobial properties of sophorolipids in medical-grade silicone discs using strains of clinical relevance. Sophorolipids were produced under fed batch conditions, ESI-MS analyses were carried out to confirm the congeners present in each formulation. Three different products were obtained SLA (acidic congeners), SL18 (lactonic congeners) and SLV (mixture of acidic and lactonic congeners) and were tested against Staphylococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC 10145 and Candida albicans IHEM 2894. All three congener mixtures showed a biofilms disruption effect (> 0.1 % w/v) of 70 %, 75 % and 80 % for S. aureus, P. aeruginosa and C. albicans, respectively. On pre-coated silicone discs, biofilm formation of S. aureus was reduced by 75 % using SLA 0.8 % w/v. After 1.5 h the inhibition of C. albicans attachment was between 45-56 % whilst after 24 h incubation the percentage of inhibition for the cell attachment increased to 68-70 % when using SLA 0.8 % w/v. Finally, in co-incubation experiments SLA 0.05 % w/v significantly reduced the ability of S. aureus and C. albicans to form biofilms and to adhere to surfaces by 90-95 % at concentrations between 0.025-0.1 % w/v. In conclusion sophorolipids significantly reduced the cell attachment of both tested strains which suggests that these molecules could have a potential role as coating agents on medical grade silicone devices for the preventions of Gram positive bacteria and yeast infections.

Autophagy induction in atrophic muscle cells requires ULK1 activation by TRIM32 through unanchored K63-linked polyubiquitin chains.

Optimal autophagic activity is crucial to maintain muscle integrity, with either reduced or excessive levels leading to specific myopathies. LGMD2H is a muscle dystrophy caused by mutations in the ubiquitin ligase TRIM32, whose function in muscles remains not fully understood. Here, we show that TRIM32 is required for the induction of muscle autophagy in atrophic conditions using both in vitro and in vivo mouse models. Trim32 inhibition results in a defective autophagy response to muscle atrophy, associated with increased ROS and MuRF1 levels. The proautophagic function of TRIM32 relies on its ability to bind the autophagy proteins AMBRA1 and ULK1 and stimulate ULK1 activity via unanchored K63-linked polyubiquitin. LGMD2H-causative mutations impair TRIM32's ability to bind ULK1 and induce autophagy. Collectively, our study revealed a role for TRIM32 in the regulation of muscle autophagy in response to atrophic stimuli, uncovering a previously unidentified mechanism by which ubiquitin ligases activate autophagy regulators.

Human PRDM2: Structure, function and pathophysiology.

PRDM2/RIZ is a member of a superfamily of histone/protein methyltransferases (PRDMs), which are characterized by the conserved N-terminal PR domain, with methyltransferase activity and zinc finger arrays at the C-terminus. Similar to other family members, two main protein types, known as RIZ1 and RIZ2, are produced from the PRDM2 locus differing by the presence or absence of the PR domain. The imbalance in their respective amounts may be an important cause of malignancy, with the PR-positive isoform commonly lost or downregulated and the PR-negative isoform always being present at higher levels in cancer cells. Interestingly, the RIZ1 isoform also represents an important target of estradiol action downstream of the interaction with hormone receptor. Furthermore, the imbalance between the two products could also be a molecular basis for other human diseases. Thus, understanding the molecular mechanisms underlying PRDM2 function could be useful in the pathophysiological context, with a potential to exploit this information in clinical practice.

Effect of biosurfactants on Pseudomonas aeruginosa and Staphylococcus aureus biofilms in a BioFlux channel.

Recent studies have indicated that biosurfactants play a role both in maintaining channels between multicellular structures in biofilms and in dispersal of cells from biofilms. A combination of caprylic acid (0.01 % v/v) together with rhamnolipids (0.04 % v/v) was applied to biofilms of Pseudomonas aeruginosa ATCC 15442, Staphylococcus aureus ATCC 9144 and a mixed culture under BioFlux flowthrough conditions and caused disruption of the biofilms. The biofilms were also treated with a combination of rhamnolipids (0.04 % v/v) and sophorolipids (0.01 %). Control treatments with PBS 1× had no apparent effect on biofilm disruption. The Gram-positive bacterium (S. aureus ATCC 9144) was more sensitive than P. aeruginosa ATCC 15442 in terms of disruption and viability as shown by Live/Dead staining. Disruption of biofilms of P. aeruginosa ATCC 15442 was minimal. Oxygen consumption by biofilms, after different treatments with biosurfactants, confirms that sophorolipid on its own is unable to kill/inhibit cells of P. aeruginosa ATCC 15442, and even when used in combination with rhamnolipids, under static conditions, no decrease in the cell viability was observed. Cells in biofilms exposed to mono-rhamnolipids (0.04 % v/v) showed behaviour typical of exposure to bacteriostatic compounds, but when exposed to di-rhamnolipids (0.04 % v/v), they displayed a pattern characteristic of bactericidal compounds.

Pseudomonas aeruginosa biofilm disruption using microbial surfactants.

AIMS: To establish the ability of the rhamnolipids biosurfactants from Pseudomonas aeruginosa, in the presence and absence of caprylic acid and ascorbic acid, to disrupt bacterial biofilms, compared with the anionic alkyl sulphate surfactant Sodium dodecyl sulphate (SDS). METHODS AND RESULTS: Pseudomonas aeruginosa ATCC 15442 biofilms were disrupted by rhamnolipids at concentrations between 0·5 and 0·4 g l(-1) and with SDS at 0·8 g l(-1) . The combination of rhamnolipids 0·4 g l(-1) and caprylic acid at 0·1 g l(-1) showed a remarkable effect on biofilm disruption and cell killing. After 30 min of treatment most of the biofilm was disrupted and cell viability was significantly reduced. Neither caprylic acid nor ascorbic acid has any effect on biofilm disruption at 0·1 g l(-1) . SDS is an effective antimicrobial agent; however, in the presence of caprylic acid its effect was neutralized. CONCLUSIONS: The results show that rhamnolipids at low concentration in the presence of caprylic acid are promising molecules for inhibition/disruption of biofilms formed by Ps. aeruginosa ATCC 15442. SIGNIFICANCE AND IMPACT OF THE STUDY: The disruption of biofilms has major significance in many industrial and domestic cleaning applications and in medical situations.

[B-mode sonography visualizing microemboli flow in the main cerebral arteries]. / Visualisation du passage de micro-emboles cérébraux par ultrasonographie en mode B-Flow.

In a patient with a mechanical prosthetic aortic valve admitted for transient amnesia, transcranial duplex Doppler and B-mode sonography visualized the transit of microemboli along the main cerebral arteries. Gaseous microemboli resulting from a cavitation phenomenon at valve closure were seen as high-intensity transient signals (HITS). To our knowledge, this is the first report of microemboli flow visualized in B-mode.

Blood pressure changes after high- and low-salt diets: are intermittent arm measures and beat-by-beat finger measures equivalent?

The assessment of sodium sensitivity requires to measure the difference in mean arterial pressure (MAP) at the end of sodium-loading (SLoad) and sodium-depletion (SDepl) maneuvers with an arm-cuff manometer. Aim of this study is to evaluate whether MAP measuring devices based on the volume-clamp method at the finger can also be used for assessing sodium sensitivity. Sixty-eight normotensive volunteers underwent SLoad and SDepl diets in random order. MAP was simultaneously measured at the end of each diet with arm (Spacelabs 90207) and finger (Portapres model-2) cuff devices. The sodium sensitivity was assessed as the difference in MAP at the end of SLoad and SDepl diets (ΔMAP), and as salt-sensitivity index (SSI; SSI = ΔMAP divided by the difference in urinary-sodium-excretion rate at the end of the diets). Discrepancies between finger and arm-cuff devices in ΔMAP or SSI were evaluated by Bland and Altman analysis. Even if discrepancies between devices had null-fixed bias, results showed a significant proportional bias and large limits of agreement (between -25 and 25 mm Hg for ΔMAP, between -196 and 180 mm Hg mol(-1) per day for SSI). The SSI distribution over the group was larger, flatter and less symmetric if derived from finger-cuff rather than arm-cuff devices, and this influenced substantially the identification of salt-sensitive individuals. Therefore, the response of MAP to SLoad/SDepl diets and consequently the assessment of the salt-sensitivity condition depends importantly on the measurement site, and brachial measures should be preferred for consistency with literature and normative data.

ICF-based approach to evaluating functionality in cardiac rehabilitation patients after heart surgery.

BACKGROUND: Heart surgery is a frequent reason for admission to in-patient cardiac rehabilitation programmes. ICF approach has never been used to evaluate cardiac patients after major heart surgery. AIM: The aim was to evaluate and measure functionality in cardiac patients who have undergone heart surgery, using for the first time the ICF-based approach and to assess whether such approach can be feasible and useful in cardiac rehabilitation. DESIGN: Observational study. SETTING: In-patients cardiac Rehabilitation Unit in Milan. POPULATION: Fifty consecutively admitted patients who had undergone heart surgery (34 males, 16 females; mean age 65.7±12.5 years). METHODS: We prepared a ICF-core set short enough to be feasible and practical. Patients were individually interviewed by different healthcare professionals (randomly selected from a group of two physicians, two physiotherapists and two psychologists) at the beginning (T1) and end of cardiac rehabilitation (T2) RESULTS: The sum of the scores of each ICF body function, body structure, activity and participation code significantly decreased between T1 and T2 (P<0.001). The environmental code scores significantly decreased in the case of facilitators between T1 and T2 (P=0.0051), but not in the case of barriers. There were significant correlations between the ICF body function scores and Barthel's index (ρ=0.381; P=0.006), NYHA class (ρ=0.404; P=0.004) and plasma Cr-P levels (r=0.31; P=0.03), between the ICF body structure codes and the Conley scale (ρ=0.306; P=0.02), and between the activity/participation codes and SpO2 (ρ=0.319; P=0.04). There were no correlations between the ICF environmental codes and clinical parameters. CONCLUSION: The ICF-based data provided functional information that was consistent with the patients' clinical course. CLINICAL REHABILITATION IMPACT: The core set used allowed to quantify important body functions and activities, including some areas that are generally insufficiently considered by healthcare professionals during cardiac rehabilitation, and document their improvement.

Wearable seismocardiography: towards a beat-by-beat assessment of cardiac mechanics in ambulant subjects.

Seismocardiogram (SCG) is the measure of the micro-vibrations produced by the heart contraction and blood ejection into the vascular tree. Over time, a large body of evidence has been collected on the ability of SCG to reflect cardiac mechanical events such as opening and closure of mitral and aortic valves, atrial filling and point of maximal aortic blood ejection. We recently developed a smart garment, named MagIC-SCG, that allows the monitoring of SCG, electrocardiogram (ECG) and respiration out of the laboratory setting in ambulant subjects. The present pilot study illustrates the results of two different experiments performed to obtain a first evaluation on whether a dynamical assessment of indexes of cardiac mechanics can be obtained from SCG recordings obtained by MagIC-SCG. In the first experiment, we evaluated the consistency of the estimates of two indexes of cardiac contractility, the pre-ejection period, PEP, and the left ventricular ejection time, LVET. This was done in the lab, by reproducing an experimental protocol well known in literature, so that our measures derived from SCG could have been compared with PEP and LVET reference values obtained by traditional techniques. Six healthy subjects worn MagIC-SCG while assuming two different postures (supine and standing); PEP was estimated as the time interval between the Q wave in ECG and the SCG wave corresponding to the opening of aortic valve; LVET was the time interval between the SCG waves corresponding to the opening and closure of the aortic valve. The shift from supine to standing posture produced a significant increase in PEP and PEP/LVET ratio, a reduction in LVET and a concomitant rise in the LF/HF ratio in the RR interval (RRI) power spectrum. These results are in line with data available in literature thus providing a first support to the validity of our estimates. In the second experiment, we evaluated in one subject the feasibility of the beat-by-beat assessment of LVET during spontaneous behavior. The subject was continuously monitored by the smart garment from 8 am to 8 pm during a workday. From the whole recording, three data segments were selected: while the subject was traveling to work (M1), during work in the office (O) and while traveling back home (M2). LVET was estimated on a beat-by-beat basis from SCG and the RRI influence was removed by regression analysis. The LVET series displayed marked beat-by-beat fluctuations at the respiratory frequency. The amplitude of these fluctuations changed in the three periods and was lower when the LF/HF RRI power ratio was higher, at O, thus suggesting a possible influence of the autonomic nervous system on LVET short-term variability. To the best of our knowledge this case report provides for the first time a representation of the beat-by-beat dynamics of a systolic time interval during daily activity. The statistical characterization of these findings remains to be explored on a larger population.

Identification of valid reference housekeeping genes for gene expression analysis in tumor neovascularization studies

INTRODUCTION: Real time RT-PCR is a widely used technique to evaluate and confirm gene expression data obtained in different cell systems and experimental conditions. However, there are many conflicting reports about the same gene or sets of gene expression. A common method is to report the interest gene expression relative to an internal control, usually a housekeeping gene (HKG), which should be constant in cells independently of experimental conditions. MATERIALS AND METHODS: In this study, the expression stability of ten HKGs was considered in parallel in two cell systems (endothelial and osteosarcoma cells): beta actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), TATA box binding protein (TBP), hypoxanthine phosphoribosyl-transferase 1 (HPRT1), Cyclophilin A (PPIA), beta-2-microglobulin (B2M), glucuronidase beta (GUSB), eukaryotic translation elongation factor 1 alpha1 (EEF1A1), transferrin receptor (TFRC), ribosomal protein S18 (RPS18). In order to study the stability of candidate reference genes, data have been also analyzed by several algorithms (geNorm, NormFinder, BestKeeper and delta-Ct method). RESULTS AND CONCLUSIONS: The overall analysis obtained by the comprehensive ranking showed that RPS18 and PPIA are appropriate internal reference genes for tumor neovascularization studies where it is necessary to analyze both systems at the same time (AU)

K-nearest-neighbor conditional entropy approach for the assessment of the short-term complexity of cardiovascular control.

Complexity analysis of short-term cardiovascular control is traditionally performed using entropy-based approaches including corrective terms or strategies to cope with the loss of reliability of conditional distributions with pattern length. This study proposes a new approach aiming at the estimation of conditional entropy (CE) from short data segments (about 250 samples) based on the k-nearest-neighbor technique. The main advantages are: (i) the control of the loss of reliability of the conditional distributions with the pattern length without introducing a priori information; (ii) the assessment of complexity indexes without fixing the pattern length to an arbitrary low value. The approach, referred to as k-nearest-neighbor conditional entropy (KNNCE), was contrasted with corrected approximate entropy (CApEn), sample entropy (SampEn) and corrected CE (CCE), being the most frequently exploited approaches for entropy-based complexity analysis of short cardiovascular series. Complexity indexes were evaluated during the selective pharmacological blockade of the vagal and/or sympathetic branches of the autonomic nervous system. We found that KNNCE was more powerful than CCE in detecting the decrease of complexity of heart period variability imposed by double autonomic blockade. In addition, KNNCE provides indexes indistinguishable from those derived from CApEn and SampEn. Since this result was obtained without using strategies to correct the CE estimate and without fixing the embedding dimension to an arbitrary low value, KNNCE is potentially more valuable than CCE, CApEn and SampEn when the number of past samples most useful to reduce the uncertainty of future behaviors is high and/or variable among conditions and/or groups.

Short-term complexity indexes of heart period and systolic arterial pressure variabilities provide complementary information.

It is unclear whether the complexity of the variability of the systolic arterial pressure (SAP) provides complementary information to that of the heart period (HP). The complexity of HP and SAP variabilities was assessed from short beat-to-beat recordings (i.e., 256 cardiac beats). The evaluation was made during a pharmacological protocol that induced vagal blockade with atropine or a sympathetic blockade (beta-adrenergic blockade with propranolol or central sympathetic blockade with clonidine) alone or in combination, during a graded head-up tilt, and in patients with Parkinson's disease (PD) without orthostatic hypotension undergoing orthostatic challenge. Complexity was quantified according to the mean square prediction error (MSPE) derived from univariate autoregressive (AR) and multivariate AR (MAR) models. We found that: 1) MSPE(MAR) did not provide additional information to that of MSPE(AR); 2) SAP variability was less complex than that of HP; 3) because HP complexity was reduced by either vagal blockade or vagal withdrawal induced by head-up tilt and was unaffected by beta-adrenergic blockade, HP was under vagal control; 4) because SAP complexity was increased by central sympathetic blockade and was unmodified by either vagal blockade or vagal withdrawal induced by head-up tilt, SAP was under sympathetic control; 5) SAP complexity was increased in patients with PD; and 6) during orthostatic challenge, the complexity of both HP and SAP variabilities in patients with PD remained high, thus indicating both vagal and sympathetic impairments. Complexity indexes derived from short HP and SAP beat-to-beat series provide complementary information and are helpful in detecting early autonomic dysfunction in patients with PD well before circulatory symptoms become noticeable.

24h seismocardiogram monitoring in ambulant subjects.

Sternal seismocardiogram (SCG) is the assessment of microvibrations produced by the beating heart as detected by an accelerometer positioned on the sternum. This signal reflects mechanical events of the heart contraction, including the opening and closure of mitral and aortic valves and maximal blood flow acceleration. Traditionally, SCG has been detected in a laboratory setting with the subject lying at rest in supine position. Aims of this study were 1) to investigate the feasibility of a SCG monitoring over the 24 hours in ambulant subjects, and 2) to calculate number and time distribution of the SCG estimates obtainable over the 24 hours. In 5 healthy subjects ECG, respiration, body accelerations and sternal SCG were recorded for 24 hours in a workday by a smart garment recently developed in our laboratory, the MagIC-SCG system. Each recording was split into a series of contiguous 5-s data segments and SCG was estimated in each segment where the magnitude of the acceleration vector was < 4 milli-g (this condition indicates that the subject was not moving).All the 24-h recordings were found of good quality and could be entirely analyzed. A large number of SCG estimates could be obtained over the 24 hours. In particular, more than 100 estimates per hour were available during the day; at night this rate was three times higher.Thus our study indicates that not only the 24h SCG monitoring in daily life is feasible but also that possible changes over time in SCG and its derived parameters may be tracked with an extreme temporal detail.

The sequence technique revised: additional concepts on the assessment of spontaneous baroreflex function.

The sequence technique is commonly employed to estimate the baroreflex sensitivity from the analysis of systolic blood pressure (SBP) and RR Interval (RRI) recordings. Traditionally, the RRI+/SBP- and RRI-/SBP+ sequences are excluded from this analysis.

Linear and fractal heart rate dynamics during sleep at high altitude. Investigation with textile technology.

OBJECTIVES: To investigate the effects of hypoxia during sleep on linear and self-similar components of heart rate variability (HRV) in eight healthy subjects at high altitude on Mount Everest. METHODS: ECG was monitored by using an innovative textile-based device, the MagIC system. For each subject three night recordings were performed at sea level (SL), at 3500 m and 5400 m above SL. RR Interval (RRI) was derived on a beat-by-beat basis from the ECG and the VLF, LF and HF spectral components and the LF/HF ratio were estimated. Short- (α1) and long-term (α2) scale exponents as well as the recently proposed spectrum of self-similarity coefficients, α(n) were estimated by detrended fluctuation analysis (DFA). RESULTS: With respect to SL, all HRV parameters but one (α2) were significantly modified at 3500 m. However, at 5400 m they tended to return to the SL values and this was in contrast with the increase in the hypobaric hypoxia and in the number of central sleep apneas occurring at higher altitude. The only HRV index that displayed changes at 5400 m was the DFA α(n) spectrum, with α(n) values significantly lower than at SL for 20 < n <50 and higher for 200 < n <400, being n the box size. CONCLUSIONS: While the biological interpretation of these results is still in progress, our data indicates that the cardiac response to high altitude hypoxia during sleep can hardly be fully explored by traditional HRV estimators only, and requires the additional support of more sophisticated indexes exploring also nonlinear and fractal features of cardiac variability.

Baroreflex contribution to blood pressure and heart rate oscillations: time scales, time-variant characteristics and nonlinearities.

The aim of this paper is to highlight the aspects of the baroreflex control of the cardiovascular system that could be relevant to the analysis and modelling of cardiovascular oscillations and regulation. In particular, complex and/or controversial issues of the baroreflex control are addressed on the basis of results obtained in previous studies by others as well as by our group. Attention has been focused on time-variant and nonlinear characteristics of the baroreflex function and on the influence of this physiological mechanism on different frequency regions of blood pressure and heart rate spectra.

Upgraded diagnostic value of Gen-Probe PACE 2 assay for detection of Chlamydia trachomatis infection.

In this study, we evaluate the performance of a nucleic acid amplification assay, COBAS AMPLICOR (Roche Molecular systems) (PCR), compared to non-amplified DNA probe assay PACE2 (Gen-Probe Inc.) for the detection of C. trachomatis in a total of 2,916 samples (2,114 females and 802 males) consecutively collected in two different clinical pathology laboratories, over a period of three years. In the females, the endocervical swabs showed a similar range of detection when using the two different methods: out of 1,581 females processed with PACE 2, 1.4% (2005), 0.9% (2006), 0.5% (2007), resulted positive for C. trachomatis; out of 533 females processed with PCR, 1.3% (2005), 1.5% (2006) and 1.2% (2007), resulted positive. However, in the male subjects we found an increased positivity of Chlamydia detection on urethral swabs by using PACE 2: 4.8% (2005), 1.9% (2006) and 2.9% (2007), compared to urine specimen processed by PCR: 1% (2005), 1.4% (2006) and 0% (2007). Even if PCR should be considered a most promising tool for routine diagnosis of Chlamydia infection, Gen Probe allowed us to better identify Chlamydia trachomatis (in 4.8% of urethral swabs compared to urine) leading to a hypothesis that extracellular EB forms of Chlamydia could be absent in urine in persistent infectious.