Inter-relationships among psychopathology, cognition, and real-life functioning in early and late phase schizophrenia: A network analysis approach.

Many illness-related factors contribute to the reduction of the real-life functioning observed in people with schizophrenia (SZ). These include the psychopathological dimensions of the disorder such as positive, negative, disorganization, and depressive symptoms as well as impairment in neurocognition, social cognition, and metacognition. The associations between some of these variables change with the duration of illness (DOI), but this aspect was not explored with a network approach. This study aimed at describing and comparing the inter-relationships between psychopathological, cognitive, and functioning variables in early (DOI ≤ 5 years) and late (DOI > 5 years) phase SZ with network analyses and at assessing which variables were more strictly and directly associated with the real-life functioning. A network representation of the relationships between variables and the calculation of centrality indices were performed within each group. The two groups were compared with a network comparison test. Seventy-five patients with early and ninety-two with late phase SZ were included. No differences in the global network structure and strength were found between the two groups. In both groups, visual learning and disorganization exhibited high centrality indices and disorganization, negative symptoms, and metacognition were directly and strongly associated with real-life functioning. In conclusion, regardless of the DOI, a rehabilitation aimed at improving visual learning and disorganization (i.e., the most central variables) might reduce the strength of the associations that compose the network and therefore indirectly facilitate functional recovery. Simultaneously, therapeutic interventions targeting disorganization and metacognition might directly improve real-life functioning.

Polymeric micelles in drug delivery: An insight of the techniques for their characterization and assessment in biorelevant conditions.

Polymeric micelles, i.e. aggregation colloids formed in solution by self-assembling of amphiphilic polymers, represent an innovative tool to overcome several issues related to drug administration, from the low water-solubility to the poor drug permeability across biological barriers. With respect to other nanocarriers, polymeric micelles generally display smaller size, easier preparation and sterilization processes, and good solubilization properties, unfortunately associated with a lower stability in biological fluids and a more complicated characterization. Particularly challenging is the study of their interaction with the biological environment, essential to predict the real in vivo behavior after administration. In this review, after a general presentation on micelles features and properties, different characterization techniques are discussed, from the ones used for the determination of micelles basic characteristics (critical micellar concentration, size, surface charge, morphology) to the more complex approaches used to figure out micelles kinetic stability, drug release and behavior in the presence of biological substrates (fluids, cells and tissues). The techniques presented (such as dynamic light scattering, AFM, cryo-TEM, X-ray scattering, FRET, symmetrical flow field-flow fractionation (AF4) and density ultracentrifugation), each one with their own advantages and limitations, can be combined to achieve a deeper comprehension of polymeric micelles in vivo behavior. The set-up and validation of adequate methods for micelles description represent the essential starting point for their development and clinical success.

inPentasomes: An innovative nose-to-brain pentamidine delivery blunts MPTP parkinsonism in mice.

Preclinical and clinical evidences have demonstrated that astroglial-derived S100B protein is a key element in neuroinflammation underlying the pathogenesis of Parkinson's disease (PD), so much as that S100B inhibitors have been proposed as promising candidates for PD targeted therapy. Pentamidine, an old-developed antiprotozoal drug, currently used for pneumocystis carinii is one of the most potent inhibitors of S100B activity, but despite this effect, is limited by its low capability to cross blood brain barrier (BBB). To overcome this problem, we developed a non-invasive intranasal delivery system, chitosan coated niosomes with entrapped pentamidine (inPentasomes), in the attempt to provide a novel pharmacological approach to ameliorate parkinsonism induced by subchronic MPTP administration in C57BL-6 J mice. inPentasomes, prepared by evaporation method was administered daily by intranasal route in subchronic MPTP-intoxicated rodents and resulted in a dose-dependent manner (0.001-0.004 mg/kg) capable for a significant Tyrosine Hydroxylase (TH) positive neuronal density rescue in both striatum and substantia nigra of parkinsonian mice. In parallel, inPentasomes significantly decreased the extent of glial-related neuroinflammation through the reduction of specific gliotic markers (Iba-1, GFAP, COX-2, iNOS) with consequent PGE2 and NO2- release reduction, in nigrostriatal system. inPentasomes-mediated S100B inhibition resulted in a RAGE/NF-κB pathway downstream inhibition in the nigrostriatal circuit, causing a marked amelioration of motor performances in intoxicated mice. On the basis of our results, chitosan coated niosomes loaded with pentamidine, the inPentasome system, self-candidates as a promising new intranasal approach to mitigate parkinsonism in humans and possibly paves the way for a possible clinical repositioning of pentamidine as anti-PD drug.

Directional K+ channel insertion in a single phospholipid bilayer: Neutron reflectometry and electrophysiology in the joint exploration of a model membrane functional platform.

We investigated the insertion of small potassium (K+) channel proteins (KcvMA-1D and KcvNTS) into model membranes and the lipid-protein structural interference, combining neutron reflectometry and electrophysiology. Neutron reflectometry experiments showed how the transverse structure and mechanical properties of the bilayer were modified, upon insertion of the proteins in single model-membranes, either supported on solid substrate or floating. Parallel electrophysiology experiments were performed on the same channels reconstituted in free-standing planar lipid bilayers, of both typical composition and matched to the neutron reflectometry experiment, assessing their electrical features. Functional and structural results converge in detecting that the proteins, conical in shape, insert with a directionality, cytosolic side first. Our work addresses the powerful combination of the two experimental approaches. We show here that membrane structure spectroscopy and ion channel electrophysiology can become synergistic tools in the analysis of structural-functional properties of biomimetic complex environment.

Water response to ganglioside GM1 surface remodelling.

BACKGROUND: Gangliosides are biological glycolipids participating in rafts, structural and functional domains of cell membranes. Their headgroups are able to assume different conformations when packed on the surface of an aggregate, more lying or standing. Switching between different conformations is possible, and is a collective event. Switching can be induced, in model systems, by concentration or temperature increase, then possibly involving ganglioside-water interaction. In the present paper, the effect of GM1 ganglioside headgroup conformation on the water structuring and interactions is addressed. METHODS: Depolarized Rayleigh Scattering, Raman Scattering, Quasielastic Neutron Scattering and NMR measurements were performed on GM1 ganglioside solutions, focusing on solvent properties. RESULTS: All used techniques agree in evidencing differences in the structure and dynamics of solvent water on different time-and-length scales in the presence of either GM1 headgroup conformations. CONCLUSIONS: In general, all results indicate that both the structural properties of solvent water and its interactions with the sugar headgroups of GM1 respond to surface remodelling. The extent of this modification is much higher than expected and, interestingly, ganglioside headgroups seem to turn from cosmotropes to chaotropes upon collective rearrangement from the standing- to the lying-conformation. SIGNIFICANCE: In a biological perspective, water structure modulation could be one of the physico-chemical elements contributing to the raft strategy, both for rafts formation and persistence and for their functional aspects. In particular, the interaction with approaching bodies could be favoured or inhibited or triggered by complex-sugar-sequence conformational switch. This article is part of a Special Issue entitled "Science for Life" Guest Editor: Dr. Austen Angell, Dr. Salvatore Magazù and Dr. Federica Migliardo.

Neutrons for rafts, rafts for neutrons.

The determination of the structure of membrane rafts is a challenging issue in biology. The selection of membrane components both in the longitudinal and transverse directions plays a major role as it determines the creation of stable or tunable platforms that host interactions with components of the outer environment. We focus here on the possibility to apply neutron scattering to the study of raft mimics. With this aim, we realized two extreme experimental models for the same complex membrane system (phospholipid : cholesterol : ganglioside GM1), involving two of the characteristic components of glycolipid-enriched rafts. One consists of a thick stack of tightly packed membranes, mixed and symmetric in composition, deposited on a silicon wafer and analyzed by neutron diffraction. The other consists of a free floating individual membrane, mixed and asymmetric in composition in the two layers, studied by neutron reflection. We present here results on the ganglioside-cholesterol coupling. Ganglioside GM1 is found to force the redistribution of cholesterol between the two layers of the model membranes. This causes cholesterol exclusion from compositionally symmetric ganglioside-containing membranes, or, alternatively, asymmetric cholesterol enrichment in raft-mimics, where gangliosides reside into the opposite layer.

Ganglioside GM1 forces the redistribution of cholesterol in a biomimetic membrane.

Neutron reflectivity has been applied to investigate different mixed asymmetric lipid systems, in the form of single "supported+floating" bilayers, made of phospholipids, cholesterol and GM1 ganglioside (Neu5Acα2-3(Galß1-3GalNAcß1-4)Galß1-4Glcß1Cer)) in bio-similar mole ratios. Bilayer preparation was carried out layer-by-layer with the Langmuir-Blodgett Langmuir-Schaefer techniques, allowing for compositional asymmetry in the system buildup. It is the first time that such a complex model membrane system is reported. Two important conclusions are drawn. First, it is experimentally shown that the presence of GM1 enforces an asymmetry in cholesterol distribution, opposite to what happens for a GM1-free membrane that, submitted to a similar procedure, results in a full symmetrization of cholesterol distribution. We underline that natural cholesterol has been used. Second, and most interesting, our results suggest that a preferential asymmetric distribution of GM1 and cholesterol is attained in a model membrane with biomimetic composition, revealing that a true coupling between the two molecular species occurs.

Lamellar stacking split by in-membrane clustering of bulky glycolipids.

We developed a simple model to investigate the effect of lipid clustering on the local interlayer distance in a cluster of interacting lamellae. The model, based on nonequilibrium thermodynamics and linear stability theories, explores the early stages of the lamella-lamella phase separation process where the lateral diffusion is much faster than the interlamellar lipid exchange. Results indicate, in the early stages, the presence of locally distorted regions with a higher concentration of one lipid component and an anomalous repeat distance. Experimental cases are presented, consisting of multilamellar-oriented depositions of phospholipids containing minority amounts of ganglioside or sphingomyelin under a low-hydration condition. The minority components are known to form domains within the phospholipid bilayer matrix. The low water content inhibits the lipid exchange among nearby lamellae and strengthens lamella-lamella interaction, allowing for a straightforward comparison with the model. Small-angle and wide-angle neutron diffraction experiments were performed in order to detect interlayer distances and local chain order, respectively. Lamellar stacking splitting has been observed for the ganglioside-containing lamellae, induced by in-phase lipid clustering. In excess water and after long equilibration times, these local structures may further evolve, leading to coexisting lamellar phases with different lipid compositions and interlayer distances.

Intermicellar interactions may induce anomalous size behavior in micelles carrying out bulky heads with multiple spatial arrangements.

We report experimental and theoretical results on the concentration dependence of the micellar size of GM1 and GM1acetyl gangliosides, five-sugar-headed anionic glycolipids. Contrary to one of the mainstays of colloid science, that the aggregation number of amphiphile aggregates grows with concentration, an anomalous region is found at intermediate concentrations, where a sharp decrease of the aggregation number occurs. Experiments were performed by small-angle X-ray and neutron scattering (SAXS and SANS). Two models are discussed, reproducing the observed behavior of either GM1acetyl or GM1. The first one is a conventional picture of interacting micelles where a reduction in the molecular surface area, leading to an increase of the aggregate dimension, is paid to reduce intermicellar interactions: it foresees a monotonous increase of the aggregation number with concentration. The second one accounts for a conformational bistability of the bulky headgroups of GM1, modifying the amphiphilic molecular surface area and protrusion from the aggregate surface, and contributing to the inter- and intramicellar interaction balance. Energy minimization leads to a complex behavior of the aggregation number, which is consistent with the anomalous behavior of GM1.

DC(13)PC bilayers from anomalous swelling to main transition: an X-ray scattering investigation.

We have performed small-angle (SAXS) and wide-angle X-ray scattering (WAXS) measurements on the lamellar phase and on large unilamellar vesicles (LUVs) of DC(13)PC in the temperature range corresponding to the anomalous swelling regime of multibilayer systems, adjacent to the chain melting transition, and across the transition. Our SAXS measurements indicate that on cooling from the L(alpha) phase, a uniform progressive swelling of the lamellar system to anomalous distances, starting approximately 2 degrees C above the main transition, is followed by a region of coexistence, covering the width of the transition ( approximately 0.6 degrees C). Across the transition region, a progressively increasing volume fraction of gel phase with a constant P (beta') interlamellar distance coexists with a decreasing amount of nongel phase that keeps on swelling to longer distances. Along both the swelling and the transition regions, anomalies in the specific heat are observed revealing a two-step process. Simultaneous WAXS experiments show a progressive "density" increase along the swelling region, constituting a direct spectroscopic evidence of an "evolving membrane" approaching the transition in a bulk real system. Calorimetric and densitometric measurements on LUVs are also presented, together with WAXS results, that show the existence of a double step main transition in a single component nanosized closed bilayer.

Age at diagnosis of deaf babies: a retrospective analysis highlighting the advantage of newborn hearing screening.

OBJECTIVE: Aim of the study was to assess the mean age at diagnosis of bilateral congenital hearing loss in the Audiology and Phoniatry Centre of the University of Turin, pointing out, by North-West Italy experience, the role of the newborn hearing screening in anticipating the age of diagnosis. METHODS: This was a retrospective study. Forty-six congenital deaf babies were reviewed and age at diagnosis was assessed for each, taking in consideration the role of hearing loss risk factors. Eighteen babies (39%) were sent by the centres that participate to the newborn hearing screening program while 28 (61%) came for parental or pediatrician suspicion of hearing loss and for general language delay. Sixteen babies (35%) presented risk factors for hearing loss. RESULTS: The mean age of identification of severe to profound hearing loss was 20.5 months (S.D.=15.3) in the whole group; considering the group of 28 babies not screened the mean age was 29.3 months (S.D.=13.4). This value decreased to 6.8 months (S.D.=3.6) in the group which underwent screening programme. This difference was statistically significant at Student's t-test (p<0.001). The average ages of diagnosis for healthy versus high risk children were significantly different only in the group of screened babies (p<0.05). CONCLUSIONS: Childhood hearing impairment is one of the most common of congenital disorders, and even though there is a general trend of early identification, in reality age of diagnosis is as yet still too late even in developed countries. Our results show that newborn hearing screening could reduce the age at which infants with hearing loss are diagnosed and treated; this would improve speech, language, auditory outcome and the quality of parents and infant life.

Voice after supracricoid laryngectomy: subjective, objective and self-assessment data.

Supracricoid laryngectomy (SCL) is an efficient surgical procedure for the treatment of selected laryngeal carcinoma, presently being performed not only in Europe but also in North America. The functional goals of the technique are voice and swallowing without a permanent tracheostoma. Perceptual and acoustic voice characteristics after SCL have been reported by different authors, but self-assessment data together with subjective and objective data have only been reported for a small number of subjects. Twenty male subjects, with a mean age of 71 years (range: 51-82 years) who underwent a SCL at least one year before our observation, were included in the study. Each subject underwent a flexible laryngoscopy and his voice was perceptually rated using the GRBAS scale. Objective examination included: maximum phonation time (MPT), voice spectrograms and syllable diadochokinesis on a single breath. Finally, each subject assessed his own voice using the Voice Handicap Index (VHI). The mean values of the GRBAS scale were respectively 2.4, 2.6, 2.4, 0.8, 0.5, 0.8. Mean MPT was 7.5 s, while for voice spectrograms the mean value of the Yanagihara scale was 3.7. Mean syllable diadochokinesis appeared as 3.3 syllables/s. Mean value of the VHI was 29.9. Subjective and objective data show a severely dysphonic voice after SCL; self-assessment data, on the contrary, reveal only moderate functional and emotional consequences. While perceptual, aerodynamic and acoustic data are in line with previous reports, self-assessment data were less severe in our subjects compared to what appears in the literature. It is concluded that self-assessment explores a different dimension of the patient's voice and that even if a severe dysphonia is present the consequences on everyday oral communication are only moderate.

Role of videoendoscopy in phoniatrics: data from three years of daily practice.

Phoniatrics is the medical specialty involved in the management of communication and swallowing disorders. Videoendoscopy plays an important role in the assessment of various disorders in the clinical practice of phoniatrics: the voice as well as the speech and swallowing mechanisms can be analysed through this procedure. Aim of the study is to describe videoendoscopic application in daily phoniatric practice: data on 1627 participants, consecutively examined, are reported. A total of 2004 videoendoscopy examinations were performed between March 1999 and December 2002. Study population comprising 1627 patients (716 male, 911 female); age ranged from 0.6 to 97 years. The following parameters were considered: a) function to be assessed through videoendoscopy (voice, speech, swallowing, other); b) phoniatric nosological chapter in participants with a recognized disease; c) age of participant; d) occupation of participant; d) medical discipline related to disease identified. Three populations were analysed: study population (1627 subjects), subjects requiring phoniatric consultation for voice and swallowing assessment. The participants examined endoscopically required a phoniatric consultation in order to have a voice or a swallowing assessment, respectively, in 67% and 20% of the cases. In 411 out of 1095 (37.5%) voice evaluations, no disorder was identified, while in most of the swallowing assessments (93.1%), a clear dysphagic disorder was detected. A bimodal distribution appeared in the 1627 participants and in the voice population with a first peak at age 20-40 years and a second peak at age 50-70. In the swallowing population, the number of participants examined appeared to increase with age. In the swallowing population, pensioners and clerks represent almost 80% of the population; in the total population, as well as in the voice populations, pensioners as well as teachers, singers and students are well represented. In most cases (52.5%), no clear disease was present. The two areas of disease most represented were otorhinolaryngology and neurology.

Utilization of macrophyte biofilter in effluent from aquaculture: I. Floating plant

The objective of this work was to manufacture a low-cost biofilter, made of floating macrophyte (Eichhornia crassipes). Limnological studies were conducted 7 days after the macrophytes were placed in the biofilter, and continued over a period of 30 consecutive days. During rainy and dry seasons, and high production period, samples were taken three times a week. The lowest levels of nitrogen compounds were observed in the July/August period, which corresponded to lower fish production and low supply rattion in the culture ponds. During the experimental period, pH values ranged from acid to alkaline and did not oscillate with higher average values during April/May. Alkalinity and bicarbonate dominance in the medium were directly affected by pH. With respect to associated micro-fauna, among phyto-plankton Chlorophyta was the dominant group and among zooplankton, Rotifera. It is recommended that during production peaks, the aquatic plants should be replaced by small buds every 10 days

Utilization of macrophyte biofilter in effluent from aquaculture: I. Floating plant.

The objective of this work was to manufacture a low-cost biofilter, made of floating macrophyte (Eichhornia crassipes). Limnological studies were conducted 7 days after the macrophytes were placed in the biofilter, and continued over a period of 30 consecutive days. During rainy and dry seasons, and high production period, samples were taken three times a week. The lowest levels of nitrogen compounds were observed in the July/August period, which corresponded to lower fish production and low supply rattion in the culture ponds. During the experimental period, pH values ranged from acid to alkaline and did not oscillate with higher average values during April/May. Alkalinity and bicarbonate dominance in the medium were directly affected by pH. With respect to associated micro-fauna, among phyto-plankton Chlorophyta was the dominant group and among zooplankton, Rotifera. It is recommended that during production peaks, the aquatic plants should be replaced by small buds every 10 days.

Utilization of macrophyte biofilter in effluent from aquaculture: I. Floating plant

The objective of this work was to manufacture a low-cost biofilter, made of floating macrophyte (Eichhornia crassipes). Limnological studies were conducted 7 days after the macrophytes were placed in the biofilter, and continued over a period of 30 consecutive days. During rainy and dry seasons, and high production period, samples were taken three times a week. The lowest levels of nitrogen compounds were observed in the July/August period, which corresponded to lower fish production and low supply rattion in the culture ponds. During the experimental period, pH values ranged from acid to alkaline and did not oscillate with higher average values during April/May. Alkalinity and bicarbonate dominance in the medium were directly affected by pH. With respect to associated micro-fauna, among phyto-plankton Chlorophyta was the dominant group and among zooplankton, Rotifera. It is recommended that during production peaks, the aquatic plants should be replaced by small buds every 10 days.

O objetivo deste trabalho foi confeccionar um biofiltro de baixo custo constituído por macrófita flutuante (Eichhornia crassipes). Os estudos limnológicos foram realizados 7 dias depois de colocadas as macrófitas no biofiltro, durante um período de 30 dias consecutivos, com amostragens 3 vezes por semana nas épocas de chuva, seca e de alta produção de organismos cultivados. Quanto aos compostos nitrogenados, as menores concentrações foram observadas no período de jul./ago., correspondendo à época de baixa produção de peixes e baixa adição de alimento nos tanques e viveiros de cultivo. O pH manteve-se ligeiramente ácido a alcalino ao longo do período experimental, não apresentando oscilações com os maiores valores médios no período de abr./mai. Os valores de pH influenciaram diretamente a alcalinidade e a dominância de bicarbonato no meio. Quanto à microfauna associada, entre os fitoplanctônicos as Chlorophyta foram o grupo dominante e entre os zooplanctônicos foram os Rotifera. Recomenda-se, no período de alta produção, substituição das plantas aquáticas por brotos bem pequenos a cada 10 dias.

Vascular changes in psoriatic knee joint synovitis.

OBJECTIVE: To evaluate the diagnostic utility of standard arthroscopy supported by a computerized image analysis system; and to examine and quantify the macroscopic appearance of blood vessels in selected anatomical areas, comparing 2 groups of patients with PsA and RA with refractory knee joint synovitis (KJS) for vascular marking (VM) features and VM scores, as well as for the relationship between respective VM scores and local and systemic KJS disease activity indices. METHODS: Standard arthroscopy was carried out on 39 knees (20 PsA, 19 RA). Videorecordings of the examination were reanalyzed using a computer image analysis system and software. The appearance of vascular markings was assessed and separately scored for the areas of surface synovium (capsular, CVM), villous proliferation (villous, VVM), and synovium adherent to cartilage (pannus, PVM). Indices of systemic (erythrocyte sedimentation rate, ESR) and local KJS disease activity (clinical index) were obtained before arthroscopy. The morphology and scores of the distinct VM were compared between PsA and RA groups, as was the relationship between respective VM scores and ESR and KJS clinical indices. RESULTS: Distinctive VM features were observed for PsA and RA KJS in each separate synovial architecture examined. VVM and CVM scores were significantly correlated with each other in PsA knees, and were significantly higher in PsA compared with RA. In both diseases, VVM and CVM scores were not related to KJS duration or activity or to ESR values, but in RA they were directly correlated with KJS activity. Moreover, the VVM capillary feature "meandering with tight convolutions," considered unique to psoriatic skin, was observed in the synovium of 13 PsA (65%) and one RA KJS (5.5%). The mean KJS duration of the PsA group with typical VVM was significantly lower than the group without VVM (2.6 +/- 1.77 vs 9.4 +/- 8.28 yrs). CONCLUSION: Our macroscopic observations of distinct changes in VM expression in selected anatomical areas of PsA and RA KJS suggest possible pathogenetic differences between the 2 diseases. The typical morphology and higher intensity of villous vascularization, in both early and chronic disease, and the different clinical relevance of VVM scores in PsA compared with RA KJS support the potential use of vascular markings as reliable outcome measures of the PsA process in KJS.

Bronchoalveolar lavage in asthmatic children: evidence of neutrophil activation in mild-to-moderate persistent asthma.

Little information is available on cell profiles and mediator production in the lower airways of children with asthma by comparison with the adult population. To study the bronchoalveolar lavage (BAL) cell profiles and production of eosinophil cationic protein (ECP) and myeloperoxidase (MPO) in childhood bronchial asthma, a retrospective study was performed in 29 children (13 allergic asthmatic children and 16 controls). Six of the asthmatics had mild-to-moderate persistent disease and seven had intermittent asthma. The BAL cell count and ECP and MPO values of asthmatic children were compared with those from 16 controls. The asthmatic patients had higher values than controls for the total cell count (p = 0.08), for neutrophils (p = 0.02), and for ECP and MPO (p < 0.001). MPO levels (p = 0.04), neutrophil count (p = 0.06), and ECP values (p = 0.06) were higher in patients with mild-to-moderate persistent asthma than in those with intermittent asthma. Our results demonstrate that neutrophil-mediated inflammation is greater in patients with more severe asthma.