Developing human laboratory models of smoking lapse behavior for medication screening

Use of human laboratory analogues of smoking behavior can provide an efficient, cost-effective mechanistic evaluation of a medication signal on smoking behavior, with the result of facilitating translational work in medications development. Although a number of human laboratory models exist to investigate various aspects of smoking behavior and nicotine dependence phenomena, none have yet modeled smoking lapse behavior. The first instance of smoking during a quit attempt (i.e. smoking lapse) is highly predictive of relapse and represents an important target for medications development. Focusing on an abstinence outcome is critical for medication screening as the US Food and Drug Administration approval for cessation medications is contingent on demonstrating effects on smoking abstinence. This paper outlines a three-stage process for the development of a smoking lapse model for the purpose of medication screening. The smoking lapse paradigm models two critical features of lapse behavior: the ability to resist the first cigarette and subsequent ad libitum smoking. Within the context of the model, smokers are first exposed to known precipitants of smoking relapse (e.g. nicotine deprivation, alcohol, stress), and then presented their preferred brand of cigarettes. Their ability to resist smoking is then modeled and once smokers 'give in' and decide to smoke, they participate in a tobacco self-administration session. Ongoing and completed work developing and validating these models for the purpose of medication screening is discussed.     

Mechanical stretching modulates growth direction and MMP-9 release in human keratinocyte monolayer

Cells within human skin are exposed to mechanical stretching that is considered a trigger stimulus for keratinocyte proliferation, while its effect on keratinocyte migration has been poorly investigate. In order to explore the effect of stretching on keratinocyte migration spontaneously immortalized human keratinocyte (HaCaT) monolayers seeded onto collagen I-coated silicon sheets were stimulated three times for 1 hour every 24 hours (total time = 72 hours) by mechanical stretching increasing substrate deformations (10%) applied both as static (0 Hz) and cyclic (0.17 Hz) uniaxial stretching. At the end of stimulations monolayer areas measured in both static and cyclic samples appeared reduced and strongly oriented in a direction perpendicular to the stress direction compared to unstimulated ones. Moreover during the mechanical stimulation period HaCaT monolayers strongly increased the release in the medium of matrix metalloproteinase 9 (MMP-9), a proteolytic enzyme necessary for keratinocyte migration.Key words: keratinocyte, mechanical stretching, migration, MMP-9, MMP-2     

Long range dispersal and spatial pattern formation in biological invasions

In this paper we explore the consequences of long distance dispersal in biological invasion processes through simulations using a recently developed cellular automaton model. We show that long distance dispersal generate characteristic spatial patterns with several stationary scale-invariant properties. In particular, the patterns display a main patch around the focus of spread, with a fractal border structure whose fractal dimension contains information about the main statistical properties of the dispersal mechanism. Our results are in agreement with field data of spread of invaders with long distance dispersal mechanisms.     

ACAT-1, Cav-1 and PrP expression in scrapie susceptible and resistant sheep

Scrapie is a prion disease for which no means of ante-mortem diagnosis is available. We recently found a relationship between cell susceptibility to scrapie and altered cholesterol homeostasis. In brains and in skin fibroblasts and peripheral blood mononuclear cells from healthy and scrapie-affected sheep carrying a scrapie-susceptible genotype, the levels of cholesterol esters were consistently higher than in tissues and cultures derived from animals with a scrapie-resistant genotype. Here we show that intracellular accumulation of cholesterol esters (CE) in fibroblasts derived from scrapie-susceptible sheep was accompanied by parallel alterations in the expression level of acyl-coenzymeA: cholesterol-acyltransferase (ACAT1) and caveolin-1 (Cav-1) that are involved in the pathways leading to intracellular cholesterol esterification and trafficking. Comparative analysis of cellular prion protein (PrPc) mRNA, showed an higher expression level in cells from animals carrying a susceptible genotype, with or without Scrapie. These data suggest that CE accumulation in peripheral cells, together with the altered expression of some proteins implicated in intracellular cholesterol homeostasis, might serve to identify a distinctive lipid metabolic profile associated with increased susceptibility to develop prion disease following infection.     

Microsatellite variability of Sardinian pine martens, Martes martes

The pine marten, Martes martes, is a medium-sized terrestrial carnivore associated with woodland habitats of the western Palearctic region. The present distribution area of the species also includes six islands of the western Mediterranean basin. The origin of these insular populations and their taxonomic status are still debated; their molecular characterization appears relevant for conservation purposes. To describe the genetic variability of the pine martens from Sardinia we characterized 40 insular and 14 Italian individuals at seven nuclear microsatellite loci. The identification of private alleles and the calculated F(ST) value of 0.074 revealed some genetic differentiation between the two populations, which accounts for the high percentages of correct allocation (96.39-98.80%) scored by the genotype assignment test. The presence of two distinct clusters corresponding to Sardinia and mainland Italy was further confirmed by the multivariate Factorial Correspondence Analysis of individual genotypes. Moreover, the genome of the Sardinian individuals bore signs of past demographic fluctuations, i.e. the presence of the monomorphic locus Ma-4, a lower allelic richness and a lower number of private alleles, which may derive from the combination of drift, founder effects, and human overexploitation. Anyway, if such events ever affected the Sardinian population, this is likely to have happened in the past since, according to our microsatellite data, the present-day population does not show evidence of recent bottlenecks or inbreeding, the Wilcoxon sign-rank test and the F(IS) index being not statistically significant (both P > 0.05). Based on this genetic evidence, we advance hypotheses about the distinctiveness of the Sardinian population and its significance for taxonomy and conservation.     

The role of mechanical stretching in the activation and localization of adhesion proteins and related intracellular molecules

The molecular complexity of the processes which lead to cell adhesion includes membrane and cytoskeletal proteins, involved in the focal adhesion formation, as well as signaling molecules tightly associated with the main intracellular regulatory cascades (Akt/PKB and MAPK/Erk). Dynamic environments, which create substrate deformations at determined frequencies and timing, have significant influences on adhesion mechanisms and in general in cellular behavior. In this work, we investigated the role of mechanical stretching (10% substrate deformation, 1 Hz frequency applied up to 60 min) on adhesion proteins (vinculin and focal adhesion kinase—FAK), related RhoGTPases (Rac1 and RhoA), and intracellular pathways (Akt/PKB and MAPK/Erk) in terms of activation and membrane recruitment in relation with cytoskeletal changes observed (membrane ruffling and filopodia formation). These changes are due to intracellular molecular rearrangements, acting with sequential concerted dynamics, able to modify the cytoskeletal conformation. The observed cellular response adds some important issues for better understanding the cellular behavior in environment which mimic as close as possible the physiological conditions. J. Cell. Biochem. 112: 1403–1409, 2011. © 2011 Wiley‐Liss, Inc.     

Harmonised Australian Reference Intervals for Serum PINP and CTX in Adults

Bone turnover markers (BTMs) are classified as either formation or resorption markers. Their concentrations in blood or urine of adults are considered to reflect the rate of bone remodelling and may be of use in the management of patients with bone disease. Major inter-method differences exist for BTMs, and harmonisation of methods is currently being pursued at an international level. Based on published data, this article describes age- and sex-specific Australian consensus reference intervals for adults for serum procollagen type I amino-terminal propeptide (s-PINP) and serum β-isomerised carboxy-terminal cross-linking telopeptide of type I collagen (s-CTX).     

中国内蒙古缨齿藓属(紫萼藓科)1新变种——缨齿藓菱形变种

报道了内蒙古清水河黄土丘陵地区发现的紫萼藓科1新变种——缨齿藓菱形变种[Jaffueliobryum wrightii(Sull.)Thér.var.rhombicumX.L.BaiSarula],该变种与干旱山地岩面生境中的原变种缨齿藓[Jaffueliobryum wrightii(Sull.)Thér.]相似,生境的变化导致其形态发生变化,主要表现在上部细胞菱形和细胞壁背部强烈加厚,未分化的叶上部边缘细胞、中肋横切面细胞不分化,叶片长0.7~0.8mm,毛尖长0.8~1.3mm。文中对缨齿藓及其新变种的形态学特征,分布和生境进行了描述,并提供了显微照片,另外,列出了缨齿藓属5个种的检索表。     

长爪沙鼠发情周期规律与判定方法

目的研究长爪沙鼠发情周期,揭示发情规律,优化判定方法。方法连续18 d采集50只长爪沙鼠阴道上皮脱落细胞涂片,采用角化细胞计数法研究长爪沙鼠发情周期规律。比较瑞氏染色、HE染色和直接镜检判定发情周期4个时相的优缺点。结果长爪沙鼠的发情周期有稳定型、不稳定型、假孕三种类型。其中稳定型占68.6%,发情周期为(106.3±35.0)h,可分为4个时相。4个时相角化细胞的比例分别为发情前期(13.5±7.8)%、发情期(86.7±9.9)%、发情后期(27.9±12.8)%和发情间期(3.3±2.8)%。结论角化细胞计数能准确地判定长爪沙鼠的发情周期及各个时相。直接镜检法能快速反映阴道脱落细胞的形态。     

人TRAIL基因cDNA的克隆及其在COS—7细胞中的表达

ＴＲＡＩＬ(ＴＮＦｒｅｌａｔｅｄａｐｏｐｔｏｓｉｓｉｎｄｕｃｉｎｇｌｉｇａｎｄ)是最近克隆的肿瘤坏死因子(ＴＮＦ)家族的新成员,由于它的蛋白质结构和生物学效应类似于ＦＡＳ/ＡＰＯ1Ｌ,因此,也被称为ＡＰＯ2Ｌ。在低浓度下,ＴＲＡＩＬ能迅速地诱导多种肿瘤细胞系的?..