Mechanism of transport of saquinavir-loaded nanostructured lipid carriers across the intestinal barrier

[EN] The aims of this work were (i) to evaluate the potential of nanostructured lipid carriers (NLCs) as a tool to 24 enhance the oral bioavailability of poorly soluble compounds using saquinavir (SQV), a BCS class IV drug 25 and P-gp substrate as a model drug, and (ii) to study NLC transport mechanisms across the intestinal barrier. 26 Three different NLC formulations were evaluated. SQV transport across Caco-2 monolayers was enhanced up 27 to 3.5-fold by NLCs compared to SQV suspension. M cells did not enhance the transport of NLCs loaded with 28 SQV. The size and amount of surfactant in the NLCs influenced SQV's permeability, the transcytosis pathway 29 and the efflux of SQV by P-gp. An NLC of size 247 nm and 1.5% (w/v) surfactant content circumvented P-gp 30 efflux and used both caveolae- and clathrin-mediated transcytosis, in contrast to the other NLC formulations, 31 which used only caveolae-mediated transcytosis. By modifying critical physicochemical parameters of the 32 NLC formulation, we were thus able to overcome the P-gp drug efflux and alter the transcytosis mechanism 33 of the nanoparticles. These findings support the use of NLCs approaches for oral delivery of poorly 34 water-soluble P-gp substrates.

The COMT Val158 Met polymorphism as an associated risk factor for Alzheimer disease and mild cognitive impairment in APOE 4 carriers

Background: The aim of this study is to examine the influence of the catechol-O-methyltranferase (COMT) gene (polymorphism Val158 Met) as a risk factor for Alzheimer's disease (AD) and mild cognitive impairment of amnesic type (MCI), and its synergistic effect with the apolipoprotein E gene (APOE). A total of 223 MCI patients, 345 AD and 253 healthy controls were analyzed. Clinical criteria and neuropsychological tests were used to establish diagnostic groups. The DNA Bank of the University of the Basque Country (UPV-EHU) (Spain) determined COMT Val158 Met and APOE genotypes using real time polymerase chain reaction (rtPCR) and polymerase chain reaction (PCR), and restriction fragment length polymorphism (RFLPs), respectively. Multinomial logistic regression models were used to determine the risk of AD and MCI. Results: Neither COMT alleles nor genotypes were independent risk factors for AD or MCI. The high activity genotypes (GG and AG) showed a synergistic effect with APOE epsilon 4 allele, increasing the risk of AD (OR = 5.96, 95% CI 2.74-12.94, p < 0.001 and OR = 6.71, 95% CI 3.36-13.41, p < 0.001 respectivily). In AD patients this effect was greater in women. In MCI patients such as synergistic effect was only found between AG and APOE epsilon 4 allele (OR = 3.21 95% CI 1.56-6.63, p = 0.02) and was greater in men (OR = 5.88 95% CI 1.69-20.42, p < 0.01). Conclusion: COMT (Val158 Met) polymorphism is not an independent risk factor for AD or MCI, but shows a synergistic effect with APOE epsilon 4 allele that proves greater in women with AD.

Anomalous behaviour of photorefractive grating erasure owing to existence of both negative and positive carriers

In LiNbO3:Fe, anomalous behaviour of grating erasure is observed with different wavelenghts, i.e. rapid grating erasure in the short wavelength range, which deviates from the results predicted by the electron transport band model. The deviation is related to the coexistance of electrons and holes in photorefraction, and charge-transfer process including electrons and hole has been proposed. The electron and hole contributions to photo-excitation coefficient S of the Fe centre on the wavelength.

Epitope mapping of antibodies to alpha-synuclein in LRRK2 mutation carriers, idiopathic Parkinson disease patients, and healthy controls

Alpha-synuclein (Snca) plays a major role in Parkinson disease (PD). Circulating anti-Snca antibodies has been described in PD patients and healthy controls, but they have been poorly characterized. This study was designed to assess the prevalence of anti-Snca reactivity in human subjects carrying the LRRK2 mutation, idiopathic PD (iPD) patients, and healthy controls and to map the epitopes of the anti-Snca antibodies. Antibodies to Snca were detected by ELISA and immunoblotting using purified recombinant Snca in plasma from individuals carrying LRRK2 mutations (104), iPD patients (59), and healthy controls (83). Epitopes of antibodies were mapped using recombinant protein constructs comprising different regions of Snca. Clear positive anti-Snca reactivity showed no correlation with age, sex, years of evolution, or the disability scores for PD patients and anti-Snca reactivity was not prevalent in human patients with other neurological or autoimmune diseases. Thirteen of the positive individuals were carriers of LRRK2 mutations either non-manifesting (8 out 49 screened) or manifesting (5 positive out 55), three positive (out of 59) were iPD patients, and five positive (out of 83) were healthy controls. Epitope mapping showed that antibodies against the N-terminal (a.a. 1-60) or C-terminal (a.a. 109-140) regions of Snca predominate in LRRK2 mutation carriers and iPD patients, being N122 a critical amino acid for recognition by the anti-C-terminal directed antibodies. Anti-Snca circulating antibodies seem to cluster within families carrying the LRRK2 mutation indicating possible genetic or common environmental factors in the generation of anti-Snca antibodies. These results suggest that case-controls' studies are insufficient and further studies in family cohorts of patients and healthy controls should be undertaken, to progress in the understanding of the possible relationship of anti-Snca antibodies and PD patholog

Lipid Nanoparticles as Carriers for RNAi against Viral Infections: Current Status and Future Perspectives

The efforts made to develop RNAi-based therapies have led to productive research in the field of infections in humans, such as hepatitis C virus (HCV), hepatitis B virus (HBV), human immunodeficiency virus (HIV), human cytomegalovirus (HCMV), herpetic keratitis, human papillomavirus, or influenza virus. Naked RNAi molecules are rapidly digested by nucleases in the serum, and due to their negative surface charge, entry into the cell cytoplasm is also hampered, which makes necessary the use of delivery systems to exploit the full potential of RNAi therapeutics. Lipid nanoparticles (LNP) represent one of the most widely used delivery systems for in vivo application of RNAi due to their relative safety and simplicity of production, joint with the enhanced payload and protection of encapsulated RNAs. Moreover, LNP may be functionalized to reach target cells, and they may be used to combine RNAi molecules with conventional drug substances to reduce resistance or improve efficiency. This review features the current application of LNP in RNAi mediated therapy against viral infections and aims to explore possible future lines of action in this field.

LDLR and PCSK9 Are Associated with the Presence of Antiphospholipid Antibodies and the Development of Thrombosis in aPLA Carriers

Introduction The identification of the genetic risk factors that could discriminate non-thrombotic from thrombotic antiphospholipid antibodies (aPLA) carriers will improve prognosis of these patients. Several human studies have shown the presence of aPLAs associated with atherosclerotic plaque, which is a known risk factor for thrombosis. Hence, in order to determine the implication of atherosclerosis in the risk of developing thrombosis in aPLA positive patients, we performed a genetic association study with 3 candidate genes, APOH, LDLR and PCSK9. Material & Methods For genetic association study we analyzed 190 aPLA carriers -100 with non-thrombotic events and 90 with thrombotic events-and 557 healthy controls. Analyses were performed by chi(2) test and were corrected by false discovery rate. To evaluate the functional implication of the newly established susceptibility loci, we performed expression analyses in 86 aPLA carrier individuals (43 with thrombotic manifestations and 43 without it) and in 45 healthy controls. Results Our results revealed significant associations after correction in SNPs located in LDLR gene with aPLA carriers and thrombotic aPLA carriers, when compared with healthy controls. The most significant association in LDLR gene was found between SNP rs129083082 and aPLA carriers in recessive model (adjusted P-value = 2.55 x 10(-3); OR = 2.18; 95% CI = 1.49-3.21). Furthermore, our work detected significant allelic association after correction between thrombotic aPLA carriers and healthy controls in SNP rs562556 located in PCSK9 gene (adjusted P-value = 1.03 x 10(-2); OR = 1.60; 95% CI = 1.24-2.06). Expression level study showed significantly decreased expression level of LDLR gene in aPLA carriers (P-value < 0.0001; 95% CI 0.16-2.10; SE 0.38-1.27) in comparison to the control group. Discussion Our work has identified LDLR gene as a new susceptibility gene associated with the development of thrombosis in aPLA carriers, describing for the first time the deregulation of LDLR expression in individuals with aPLAs. Besides, thrombotic aPLA carriers also showed significant association with PCSK9 gene, a regulator of LDLR plasma levels. These results highlight the importance of atherosclerotic processes in the development of thrombosis in patients with aPLA.

Modeling the development of pronunciation in infant speech acquisition.

Pronunciation is an important part of speech acquisition, but little attention has been given to the mechanism or mechanisms by which it develops. Speech sound qualities, for example, have just been assumed to develop by simple imitation. In most accounts this is then assumed to be by acoustic matching, with the infant comparing his output to that of his caregiver. There are theoretical and empirical problems with both of these assumptions, and we present a computational model- Elija-that does not learn to pronounce speech sounds this way. Elija starts by exploring the sound making capabilities of his vocal apparatus. Then he uses the natural responses he gets from a caregiver to learn equivalence relations between his vocal actions and his caregiver's speech. We show that Elija progresses from a babbling stage to learning the names of objects. This demonstrates the viability of a non-imitative mechanism in learning to pronounce.

KLAIR: A virtual infant for spoken language acquisition research

Recent research into the acquisition of spoken language has stressed the importance of learning through embodied linguistic interaction with caregivers rather than through passive observation. However the necessity of interaction makes experimental work into the simulation of infant speech acquisition difficult because of the technical complexity of building real-time embodied systems. In this paper we present KLAIR: a software toolkit for building simulations of spoken language acquisition through interactions with a virtual infant. The main part of KLAIR is a sensori-motor server that supplies a client machine learning application with a virtual infant on screen that can see, hear and speak. By encapsulating the real-time complexities of audio and video processing within a server that will run on a modern PC, we hope that KLAIR will encourage and facilitate more experimental research into spoken language acquisition through interaction. Copyright © 2009 ISCA.

EARLY MOTHER-INFANT RELATIONSHIPS OF MACACA-THIBETANA AT MT-EMEI CHINA

Quantitative data on early mother-infant relationships in the Tibetan macaque was collected during the first 23 weeks of infant life in spring, 1987, at Mt. Emei, China. During the first week of life, infants spent 98.3% of their time in ventroventral contact with their mothers. This contact rapidly decreased to 33.8% by the 4th week and thereafter to 0.85% by the 23rd week. Nipple contact decreased relatively slowly from 89.7% to 62.9% within the first 4 weeks of infant life and to 19.8% by the 23rd week. Ventrolateral and ventrodorsal contact appeared by the 2nd week, mean-while, maternal restraining behavior appeared, and reached a peak by the 3rd week. The mother neither encouraged nor discouraged her infant's independence during 4-8th weeks. Maternal rejection of the infant was first observed when the infant was 11 weeks old and continued thereafter.

Far-infrared emission and absorption by hot carriers in superlattices

Some of the earliest theoretical speculation, stimulated by the growth of semiconductor superlattices, focused on novel devices based on vertical transport through engineered band structures; Esaki and Tsu promised Bloch oscillators in narrow mini-band systems and Kazarinov and Suris contemplated electrically stimulated intersubband transitions as sources of infrared radiation. Nearly twenty years later these material systems have been perfected, characterized and understood and experiments are emerging that test some of these original concepts for novel submillimetre wave electronics. Here we describe recent experiments on intersubband emission in quantum wells stimulated by resonant tunnelling currents. A critical issue at this time is devising a way to achieve population inversion. Other experiments explore 'saturation' effects in narrow miniband transport. Thermal saturation may be viewed as a precursor to Bloch oscillation if the same effects can be induced with an applied electric field.

Male-infant-male interactions in Tibetan macaques

Event-sampling and scans were used for collecting data on male-infant-male triadic interactions, and their effects on member spacing respectively in a group of Macaca thibetana at Mt. Emei in 1989. The group was partially provisioned by human visitors in seasons other than winter, and could be observed closely. In addition, a stable linear male-hierarchy among five males existed for two years since the end of 1987, providing a good social condition for this topic. The triadic interactions were specific to the birth season, and recognized as three types being on a continuum functionally changing from passive ''agonistic buffering'' (4.8%) to active spatial cohesion, which resulted in a significant decline of intermale distances. Positive correlations were documented between the triad initiation rate and the number of females in consort with the males in the mating season (MS), and between the triad reception rate and the number of infants in proximity to the males in the MS when maternal care was significantly reduced. Thus the male's mating effort and kin/sexual selection may deeply be involved in the triad of this species. Considering that the two triad-species, M. sylvanus and M. thibetana, had different levels of paternity, but shared similar foraging conditions, and showed similar intensities of male-infant caretaking, the triad was very likely a byproduct of male-infant caretaking, which was probably shaped to compensate heavy maternal investment to young offspring in harsh conditions. Accordingly, the long-term arguments about the triad in M. sylvanus can be united to a model of the way in which ''male-infant caretaking'' hypothesis works ultimately, and ''regulating social relations'' hypothesis does proximately.