bluuebunny/crossref_metadata_2025 huggingface dataset:
Synchronous high‐grade squamous intraepithelial lesion and adenocarcinoma in situ of cervix in a young woman presenting with hyperchromatic crowded groups in the cervical cytology specimen: Report of a case
Authors: Zafar Nadeem , Balazs Louisa , Benstein Barbara D.
Published: October 2008

We report a 29‐year‐old woman who underwent routine gynecologic evaluation at a community clinic and had a cervical sample drawn for liquid‐based cytologic evaluation. At cytology, many hyperchromatic crowded groups (HCG) were present, but a consensus could not be established whether the abnormal cells were primarily glandular or squamous with secondary endocervical glandular involvement. An interpretation of atypical endocervical cells, favor neoplastic, was rendered and biopsy advised if clinically appropriate. At biopsy, the cervix contained synchronous squamous cell carcinoma in situ, secondarily involving endocervical glands, and neighboring adenocarcinoma in situ. Immunohistochemistry for Ki‐67 and p16INK4A crisply and precisely stained both the lesions, clearly separating them from the adjacent uninvolved mucosa. This case re‐emphasizes the challenge associated with accurate evaluation of HCG at cytology, the significance of ancillary testing for surrogate markers of high‐risk HPV (HR‐HPV) infection, the need for adjunct testing for HPV‐DNA in the setting of HCG at cervical cytology, and a recommendation to set up studies to evaluate the role of surrogate markers of HR‐HPV infection in cytologic samples with HCG. Diagn. Cytopathol. 2008;36:823–826. © 2008 Wiley‐Liss, Inc.
Structural and Spectroscopic Properties of New Copper(I) Complexes with 1,1,1‐Tris(diphenylphosphanylmethyl)ethane and Heterocyclic Thiolates
Authors: Aslanidis Paraskevas , Cox Philip J. , Kapetangiannis Konstantinos , Tsipis Athanassios C.
Published: October 2008

Reactions of copper(I) halides with 1,1,1‐tris(diphenylphosphanylmethyl)ethane (triphos) in 1:1 molar ratio afforded mononuclear complexes of the type [CuX(triphos)] which on further treatment with one equivalent of the potassium thiolate salt of a heterocyclic thione gave rise to the formation of mixed‐ligand complexes of the formula [Cu(κ3‐triphos)(κ1‐thiolate)]. The molecular structures of [Cu(κ3‐triphos)(κ1‐py2S)]·C2H5OH (1) and [Cu(κ3‐triphos)(κ1‐pymt)] (2) have been established by single‐crystal X‐ray diffraction. The complexes adopt a rigid distorted tetrahedral geometry with the phosphane ligand acting in a tridentate chelating mode. The complexes are strongly luminescent in solution and in the solid state. The spectroscopic and electronic properties of the new copper(I) complexes have been computed by electronic structure computational techniques (TD‐DFT calculations), that contribute to the assignments of electronic spectra. The high‐energy absorptions in the absorption spectra of complexes 1 and 2, ranging from 210 to 270 nm, are assigned to MLCT/LLCT transitions and the same also holds true for the low‐energy absorptions found in the range 280 to 300 nm. In the lowest triplet excited state, T1, involved in the emission in the 456–502 nm range, the spin density is delocalized over the entire nuclear framework.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)
Construction of periventricular white matter hyperintensity maps by spatial normalization of the lateral ventricles
Authors: Jongen Cynthia , van der Grond Jeroen , Anbeek Petronella , Viergever Max A. , Biessels Geert Jan , Pluim Josien P.W.
Published: October 2008

Subcortical and periventricular white matter hyperintensities (WMHs) may have different associations with cognition and pathophysiology. The aim of the present study is to develop an automated method for construction of periventricular WMH maps that enables the analysis of between‐group differences in WMH location and characteristics in the periventricular region without the requirement of prior boundary definition. To avoid influence of WMHs on spatial normalization, a reference image of the lateral ventricles was constructed based on images of 24 subjects. Construction was not biased to a single subject. WMHs were segmented by k‐nearest neighbor‐based classification of magnetic resonance inversion recovery and fluid attenuated inversion recovery images. Cerebrospinal fluid segmentations of individual subjects were nonrigidly mapped to the reference image of the lateral ventricles. The subject's WMHs were transformed to the reference space accordingly. Spatial normalization accuracy was validated using measures of overlap and of displacement relative to the boundary of the lateral ventricles. After spatial normalization, the boundaries of the lateral ventricles closely matched the reference image and in an area of ∼1 cm around the lateral ventricles the relative displacement was less than 1 mm. To illustrate the method, it was applied to 61 patients with Type 2 diabetes and 26 control subjects, whereupon periventricular WMH maps were constructed and compared. The proposed method is particularly suited to analyze WMH distribution differences at the level of the lateral ventricles between large groups of patients. Hum Brain Mapp, 2009. © 2008 Wiley‐Liss, Inc.
Brn‐4 is upregulated in the deafferented hippocampus and promotes neuronal differentiation of neural progenitors in vitro
Authors: Zhang Xinhua , Jin Guohua , Wang Lei , Hu Wenzhong , Tian Meiling , Qin Jianbing , Huang Huiwei
Published: October 2008

Fimbria‐fornix (FF), the septo‐hippocampal pathway, was transected to model Alzheimer's disease (AD), which is characterized by loss of cholinergic afferent fibers in hippocampus. Various alternations may happen in the deafferented hippocampus. In this study, we determined the expression of Brn‐4 in hippocampus after FF lesion. RT‐PCR and Western blot showed that mRNA transcription and protein of Brn‐4 increased significantly and reached to the peak at day 14 after FF lesion. Hybridization and immunohistochemistry indicated that Brn‐4 signals in hippocampus and dentate gyrus (DG) of the deafferented side were significantly stronger than the normal side. More Brn‐4 positive cells were identified in the DG of deafferented hippocampus. In the pyramidal and granular cells, Brn‐4 positive cells were all NeuN positive neurons, whereas in the neurogenic area, subgranular zone (SGZ), only a part of Brn‐4 positive cells were NeuN positive, and these Brn‐4/NeuN double positive neurons in SGZ and hilus of DG increased significantly after the trauma induced by FF lesion. In vitro Brn‐4 antibody attenuated the role of extract from deafferented hippocampus in promoting differentiation of hippocampal progenitors into MAP‐2 positive neurons. This study demonstrated that after FF lesion, Brn‐4 in the deafferented hippocampus was upregulated and might play an important role in inducing local progenitors to differentiate into neurons, which may compensate for the loss of cholinergic afferent fibers or other dysfunctions. © 2008 Wiley‐Liss, Inc.
Contributions of the hippocampus and medial prefrontal cortex to energy and body weight regulation
Authors: Davidson Terry L. , Chan Kinho , Jarrard Leonard E. , Kanoski Scott E. , Clegg Deborah J. , Benoit Stephen C.
Published: October 2008

The effects of selective ibotenate lesions of the complete hippocampus (CHip), the hippocampal ventral pole (VP), or the medial prefrontal cortex (mPFC) in male rats were assessed on several measures related to energy regulation (i.e., body weight gain, food intake, body adiposity, metabolic activity, general behavioral activity, conditioned appetitive responding). The testing conditions were designed to minimize the nonspecific debilitating effects of these surgeries on intake and body weight. Rats with CHip and VP lesions exhibited significantly greater weight gain and food intake compared with controls. Furthermore, CHip‐lesioned rats, but not rats with VP lesions, showed elevated metabolic activity, general activity in the dark phase of the light‐dark cycle, and greater conditioned appetitive behavior, compared with control rats without these brain lesions. In contrast, rats with mPFC lesions were not different from controls on any of these measures. These results indicate that hippocampal damage interferes with energy and body weight regulation, perhaps by disrupting higher‐order learning and memory processes that contribute to the control of appetitive and consummatory behavior. © 2008 Wiley‐Liss, Inc.
The neural correlates of recollection: Hippocampal activation declines as episodic memory fades
Authors: Viskontas Indre V. , Carr Valerie A. , Engel Stephen A. , Knowlton Barbara J.
Published: October 2008

Memories for certain events tend to linger in rich, vivid detail, and retrieval of these memories includes a sense of re‐experiencing the details of the event. Most events, however, are not retained in any detailed way for more than a few days. According to one theory, the hippocampus plays a specific role in supporting episodic retrieval, that is, the re‐experiencing of an event as part of one's personal past. This theory predicts that as episodic memories fade over time and are reduced to feelings of familiarity, activity in the hippocampus should no longer be associated with retrieval. We used high‐resolution functional imaging to explore neural activity in medial temporal lobe subregions while participants performed a recognition task at both a short (10‐min) and long (1‐week) study‐test delay. For each recognized item, subjects made “Remember/Know” judgments, allowing us to distinguish between items that were consistently episodic across the two tests and items that were initially episodic, but later became merely familiar. Our results demonstrate that activity in the subiculum is specifically associated with episodic recollection. Overall, recollected items were associated with higher activity in the subiculum than other items. For transiently recollected items, there was a decrease in subicular activity across the 1‐week delay as memory faded from recollection to familiarity, whereas consistently recollected items were associated with enhanced subicular activity at both delays. These results provide evidence of a link between subicular activation and recollective experience. © 2008 Wiley‐Liss, Inc.
Enzymatic esterification of lactic acid, utilizing the basicity of particular polar organic solvents to suppress the acidity of lactic acid
Authors: Hasegawa Satoshi , Azuma Masanori , Takahashi Koji
Published: October 2008

BACKGROUND: Lipase‐catalyzed esterification of lactic acid with ethanol was investigated; however, some difficulties exist with such a system. Because of its high polarity, lactic acid is immiscible with non‐polar organic solvents, which have generally been used for non‐aqueous enzyme reactions. In addition, the strong acidity of lactic acid causes acid inactivation of enzymes and acid‐catalyzed, non‐enzymatic esterification.

RESULT: In the present study, particular polar organic solvents, such as 1,4‐dioxane, were found to suppress the enzyme inactivation and non‐enzymatic esterification caused by the acidity. The magnitude of this effect varied with solvents and strongly correlated with the Kamlet‐Taft parameter β, which indicates the basicity of the solvents in non‐aqueous systems. An immobilized lipase from Candida antarctica was found to be the most active and stable enzyme for the reaction. Based on the findings, lipase‐catalyzed esterification of lactic acid (1.0 mol L−1) could be continued for up to 4 weeks without any loss of enzyme activity.

CONCLUSION: In addition to miscibility with lactic acid, the effect of these polar solvents, which is to suppress the acidity of lactic acid and is presumably due to the basicity, appears to play a very important role in the efficient enzymatic reaction of lactic acid. Copyright © 2008 Society of Chemical Industry
Combined permeable pavement and ground source heat pump systems to treat urban runoff
Authors: Scholz Miklas , Grabowiecki Piotr
Published: October 2008

BACKGROUND: This paper assesses the functioning of the next generation of permeable pavement systems. The overall concept is to combine traditional permeable pavement systems with ground source heat pump systems. Long‐term research to provide data concerning nutrient reductions and microbial activity within experimental pavement rigs is required.

RESULTS: The variability of temperature allowed for the survival of pathogenic organisms within the sub‐base of permeable pavement systems. Counts of microbes and nutrient concentrations have been determined by direct observation (plate count tests) and chemical analysis, respectively. An analysis using numerical methods indicated 99% and 95% biochemical oxygen demand and ammonia‐nitrogen removal, respectively. Supplementary carbon dioxide monitoring indicated relatively high microbial activity on geotextiles and within the lower parts of the sub‐base. Low oxygen concentrations were measured in the space around the geotextile, where carbon dioxide concentrations reached 2000 ppm.

CONCLUSIONS: The research enables decision‐makers for the first time to assess public health risks, treatment requirements and efficiencies, and the potential for runoff recycling. The great system stability of the innovation and minor water quality data variability between individual experimental pavement systems provide good evidence for the controlled engineered application of this novel sustainable technology. Copyright © 2008 Society of Chemical Industry
Decolourization of textile dyes by the white‐rot fungi Phanerochaete chrysosporium and Pleurotus ostreatus
Authors: Faraco V. , Pezzella C. , Giardina P. , Piscitelli A. , Vanhulle S. , Sannia G.
Published: October 2008

BACKGROUND: The ability of the fungi Pleurotus ostreatus and Phanerochaete chrysosporium to decolourize and detoxify 11 (mono‐, dis‐, poly‐ azo, and anthraquinonic type) dyes, widely used across the textile and leather industries, was tested.

RESULTS: Different substrate specificities were revealed between P. ostreatus and P. chrysosporium in decolourization experiments. The latter fungus provided almost complete decolourization of the tested azo dyes up to 600 ppm and dis‐azo dyes up to 1000 ppm, and 80% decolourization of the tris‐azo dye DBU1L38 at 1000 ppm, after 6 days. P. ostreatus provided almost total decolourization of the anthraquinone type dye ABU62 (1000 ppm) after just 1 day. P. ostreatus also showed the ability to decolourize the tested dis‐azo and tris‐azo dyes, giving the best performances against the dis‐azo DBU1U1 (600 ppm) dye, which was 100% decolourized after 6 days. Laccases proved to be the main enzymatic activities acting in P. ostreatus decolourization.

CONCLUSION: The potential of the fungi P. ostreatus and P. chrysosporium as efficient bio‐systems for decolourization and detoxification of several toxic industrial dyes was demonstrated. The role of laccases in the decolourization of dis‐azo dyes by P. ostreatus was demonstrated for the first time. Copyright © 2008 Society of Chemical Industry
Short‐term low‐salinity tolerance by the longhorn sculpin, Myoxocephalus octodecimspinosus
Authors: Hyndman Kelly A. , Evans David H.
Published: October 2008

The bottom‐dwelling, longhorn sculpin, Myoxocephalus octodecimspinosus, is traditionally viewed as a stenohaline marine fish, but fishermen have described finding this sculpin in estuaries during high tide. Little is known about the salinity tolerance of the longhorn sculpin; thus, the purposes of these experiments were to explore the effects of low environmental salinity on ion transporter expression and distribution in the longhorn sculpin gill. Longhorn sculpin were acclimated to either 100% seawater (SW, sham), 20% SW, or 10% SW for 24 or 72 hr. Plasma osmolality, sodium, potassium, and chloride concentrations were not different between the 20 and 100% treatments; however, they were 20–25% lower with exposure to 10% SW at 24 and 72 hr. In the teleost gill, regulation of Na+, K+‐ATPase (NKA), Na+–K+–2Cl− cotransporter (NKCC1), and the chloride channel, cystic fibrosis transmembrane conductance regulator (CFTR) are necessary for ion homeostasis. We immunolocalized these proteins to the mitochondrion‐rich cell of the gill and determined that acclimation to low salinity does not affect their localization. Also, there was not a downregulation of gill NKA, NKCC1, and CFTR mRNA or protein during acclimation to low salinities. Collectively, these results suggest that down to 20% SW longhorn sculpin are capable of completely regulating ion levels over a 72‐hr period, whereas 10% SW exposure results in a significant loss of ions and no change in ion transporter density or localization in the gill. We conclude that longhorn sculpin can tolerate low‐salinity environments for days but, because they cannot regulate ion transporter density, they are unable to tolerate low salinity for longer periods or enter freshwater (FW). The genus Myoxocephalus has three FW species, making this group an excellent model to test evolutionary and physiological mechanisms that allow teleosts to invade new low salinities successfully. J. Exp. Zool. 311A:45–56, 2009. © 2008 Wiley‐Liss, Inc.
Evidence of multiple paternity in Morelet's Crocodile (Crocodylus moreletii) in Belize, CA, inferred from microsatellite markers
Authors: McVay John D. , Rodriguez David , Rainwater Thomas R. , Dever Jennifer A. , Platt Steven G. , McMurry Scott T. , Forstner Michael R.J. , Densmore Llewellyn D.
Published: October 2008

Microsatellite data were generated from hatchlings collected from ten nests of Morelet's Crocodile (Crocodylus moreletii) from New River Lagoon and Gold Button Lagoon in Belize to test for evidence of multiple paternity. Nine microsatellite loci were genotyped for 188 individuals from the 10 nests, alongside 42 nonhatchlings from Gold Button Lagoon. Then mitochondrial control region sequences were generated for the nonhatchlings and for one individual from each nest to test for presence of C. acutus‐like haplotypes. Analyses of five of the nine microsatellite loci revealed evidence that progeny from five of the ten nests were sired by at least two males. These data suggest the presence of multiple paternity as a mating strategy in the true crocodiles. This information may be useful in the application of conservation and management techniques to the 12 species in this genus, most of which are threatened or endangered. J. Exp. Zool. 309A:643–648, 2008. © 2008 Wiley‐Liss, Inc.
Household targeting in practice: The Nicaraguan Red de Protección Social
Authors: Maluccio John A.
Published: October 2008

This article describes the details underlying the targeting of a Nicaraguan anti‐poverty program, emphasising the rationale for how it was designed and implemented. It offers, by way of example, a guide for targeting in an anti‐poverty program, and highlights some of the potential tradeoffs. It then goes on to present a quantitative assessment of how well the program was able to target poor households. A combination of ad hoc and statistical procedures led to targeting that was effective, with undercoverage rates of 10 per cent or below and leakage rates of 15 per cent or below. This was in spite of the fact that the targeting methodologies used were imprecise at both the household and geographic levels. Copyright © 2008 John Wiley & Sons, Ltd.
Institutions and economic performance: Implications for African development
Authors: Luiz John M.
Published: October 2008

The recent economic performance of Sub Saharan Africa has been very disappointing. Its poverty has many dimensions and causes, both internal and external. This paper focuses on the role of institutions in promoting or hindering economic development in Africa and the implications for doing business on the continent. It questions our understanding of institutions and how they develop and warns against simplistic assumptions in this regard. The paper examines how it is that institutions come to affect economic growth and the characteristics of what makes for good institutions. Copyright © 2008 John Wiley & Sons, Ltd.
Romanticising the poor harms the poor
Authors: Karnani Aneel
Published: October 2008

A libertarian movement that emphasises free markets to reduce poverty has grown strong in recent years. It views the poor as ‘resilient and creative entrepreneurs and value‐conscious consumers’. This romanticised view of the poor is far from the truth and harms the poor in two ways. First, it results in too little emphasis on legal, regulatory and social mechanisms to protect the poor who are vulnerable consumers. Second, it results in overemphasis on microcredit and under‐emphasis on fostering modern enterprises that would provide employment opportunities for the poor. More importantly, the libertarian proposition grossly under‐emphasises the critical role and responsibility of the state for poverty reduction. Copyright © 2008 John Wiley & Sons, Ltd.
On consumption insurance in poor urban areas: Evidence from Ethiopia
Authors: Alvi Eskander , Dendir Seife
Published: October 2008

This paper tests the full insurance and related hypotheses using data from urban households in Ethiopia. Whereas consumption insurance tests have focused on rural areas of developing countries, much less is known about the urban poor who remain precariously positioned in terms of consumption insurance—owing both to weak social bonding that undermines informal arrangements and thin formal insurance and financial markets. Results from OLS and GMM estimations provide evidence to this effect and show that full insurance is strongly rejected for various income and consumption categories even at the local level. The results do suggest however that partial insurance may be taking place though only among the most vulnerable. Copyright © 2008 John Wiley & Sons, Ltd.
C1q, the recognition subcomponent of the classical pathway of complement, drives microglial activation
Authors: Färber Katrin , Cheung Giselle , Mitchell Daniel , Wallis Russell , Weihe Eberhard , Schwaeble Wilhelm , Kettenmann Helmut
Published: October 2008

Microglia, central nervous system (CNS) resident phagocytic cells, persistently police the integrity of CNS tissue and respond to any kind of damage or pathophysiological changes. These cells sense and rapidly respond to danger and inflammatory signals by changing their cell morphology; by release of cytokines, chemokines, or nitric oxide; and by changing their MHC expression profile. We have shown previously that microglial biosynthesis of the complement subcomponent C1q may serve as a reliable marker of microglial activation ranging from undetectable levels of C1q biosynthesis in resting microglia to abundant C1q expression in activated, nonramified microglia. In this study, we demonstrate that cultured microglial cells respond to extrinsic C1q with a marked intracellular Ca2+ increase. A shift toward proinflammatory microglial activation is indicated by the release of interleukin‐6, tumor necrosis factor‐α, and nitric oxide and the oxidative burst in rat primary microglial cells, an activation and differentiation process similar to the proinflammatory response of microglia to exposure to lipopolysaccharide. Our findings indicate 1) that extrinsic plasma C1q is involved in the initiation of microglial activation in the course of CNS diseases with blood–brain barrier impairment and 2) that C1q synthesized and released by activated microglia is likely to contribute in an autocrine/paracrine way to maintain and balance microglial activation in the diseased CNS tissue. © 2008 Wiley‐Liss, Inc.
Involvement of calpain in the process of Jurkat T cell chemotaxis
Authors: Butler Jonathan T. , Samantaray Supriti , Beeson Craig C. , Ray Swapan K. , Banik Naren L.
Published: October 2008

Massive T cell infiltration into the central nervous system is a hallmark of multiple sclerosis (MS) and its rodent model experimental autoimmune encephalomyelitis (EAE), resulting in the induction of many of the pathophysiological events that lead to neuroinflammation and neurodegeneration. Thus, blocking T cell migration into the central nervous system may reduce disease severity in MS and EAE. One potential target for reducing T cell migration is inhibition of the Ca2+‐activated neutral protease calpain. Previous studies in other cell types have demonstrated that migration is reduced by incubation of cells with calpain inhibitors. Thus, we hypothesize that calpain inhibition will reduce migration of T cells in response to and toward the chemokine CCL2. To test this hypothesis, the intracellular free Ca2+ levels in Jurkat E6‐1 T cells was first measured by the fura‐2 assay to assess whether the intracellular ion environment would support calpain activation. The intracellular free Ca2+ levels were found to increase in response to CCL2. The cells were next treated with the calpain inhibitor calpeptin in a multiwelled Boyden chamber with CCL2 used as the chemoattractant. These studies demonstrate that inhibition of calpain with its inhibitor calpeptin produces a dose‐dependent inhibition of chemotaxis. Calpain activity, as measured by live cell imaging, was also increased in response to CCL2, providing further evidence of its involvement in the process of chemotaxis and migration. These studies provide evidence for the involvement of calpain in the mechanisms of chemotaxis and warrants further exploration in MS patient and EAE animal samples. © 2008 Wiley‐Liss, Inc.
Entrainment and coupling of the hamster suprachiasmatic clock by daily dark pulses
Authors: Mendoza Jorge , Pévet Paul , Challet Etienne
Published: October 2008

The circadian rhythm of locomotor activity of hamsters kept in constant light (LL) can split into two distinct components that, in steady state, lie 180° apart. The splitting phenomenon is the result of antiphase circadian oscillations between left and right sides of the suprachiasmatic nuclei (SCN), the master circadian clock in mammals. In unsplit hamsters housed in LL, a single dark pulse produces a phase‐shift of the wheel‐running activity rhythm, accompanied by a transient down‐regulation of clock gene expression in the SCN. In the present study, we evaluated the effects of daily 1‐hr dark pulses on wheel‐running activity rhythm and on the expression of clock and nonclock proteins in the SCN of Syrian hamsters exposed to LL conditions. The results show that a daily 1‐hr dark pulse entrained the rhythm of wheel‐running activity of unsplit hamsters. In addition, in split animals, unimodal coupling of the two locomotor activity components was produced by daily 1‐hr dark pulses. In the SCN, the effects of entrainment and unimodal coupling of the two separate components by dark observed in behavior were also evident in the bilateral expression of the proteins c‐FOS, p‐ERK, PERIOD 1, and calbindin. These results show that the bilaterally asymmetric SCN clock, underlying split circadian behavior, can be recoupled in phase and entrained by short daily dark exposure, indicating the synchronizing potency of darkness on the main circadian clock. © 2008 Wiley‐Liss, Inc.
Minocycline: A neuroprotective agent for hypoxic‐ischemic brain injury in the neonate?
Authors: Buller Kathryn M. , Carty Michelle L. , Reinebrant Hanna E. , Wixey Julie A.
Published: October 2008

Minocycline is a second‐generation tetracycline and a potential neuroprotective intervention following brain injury. However, despite the recognized beneficial effects of minocycline in a multitude of adult disease states, the clinical application of minocycline in neonates is contentious. Tetracyclines, as a class, are not usually administered to neonates, but there is compelling evidence that minocycline reduces brain injury after neonatal hypoxic‐ischemic brain injury. This Review focuses on the evidence for minocycline use in neonates by considering aspects of pharmacology, drug regimens, functional outcomes, and mechanisms of action. © 2008 Wiley‐Liss, Inc.
Monoamine oxidase‐B activity is not involved in the neuroinflammatory response elicited by a focal freeze brain injury
Authors: Sanz Elisenda , Quintana Albert , Valente Tony , Manso Yasmina , Hidalgo Juan , Unzeta Mercedes
Published: October 2008

Cryolesion of the frontoparietal cortex in mice is a well‐described brain injury paradigm that results in increased astrogliosis surrounding the lesion site and is accompanied by a prominent increase in the MAO‐B levels in astrocytes. Whether these increased MAO‐B levels contribute to cellular damage or modulate reactive astrocytosis remains unclear. MAO‐B activity may contribute to cellular damage, since its metabolism products are highly toxic to the cells. Additionally, it has been suggested that MAO‐B inhibition may regulate astrocytic reaction. In this study, we have determined the relative contribution of MAO‐B activity to the outcome following freeze injury. Freeze injury induced a prominent increase of several inflammatory markers, including ICAM, Mac‐1, EB22, and GFAP. Inhibition of MAO‐B activity using the selective inhibitor PF9601N did not reduce this cryolesion‐induced inflammatory response. Additional data revealed that the expression of several cryolesion‐induced cell death genes, such as Fas, Rip, p53, and ICE, was not reduced in PF9601N‐treated mice, evidencing that MAO‐B activity did not contribute to cryolesion‐induced cell death. Definitive functional analysis of the mice using the ladder beam task revealed that MAO‐B inhibition did not improve the cryolesion‐induced motor impairment. These data strongly suggest that, although MAO‐B is highly expressed in the area surrounding the lesion site, its activity does not contribute to the cellular damage or play any role in regulating astrocytic reactivity. © 2008 Wiley‐Liss, Inc.
Controlled contusion injury alters molecular systems associated with cognitive performance
Authors: Griesbach Grace Sophia , Sutton Richard L. , Hovda David A. , Ying Zhe , Gomez‐Pinilla Fernando
Published: October 2008

We investigated whether a learning impairment after a controlled cortical impact (CCI) injury was associated with alterations in molecules involved in synaptic plasticity and learning and memory. Adult male rats with moderate CCI to the left parietal cortex, tested in a Morris water maze (MWM) beginning at postinjury day 10, showed impaired cognitive performance compared with sham‐treated rats. Tissue was extracted for mRNA analysis on postinjury day 21. The expression of brain‐derived neurotrophic factor (BDNF), synapsin I, cyclic‐AMP response element binding protein (CREB), and calcium‐calmodulin–dependent protein kinase II (α‐CAMKII) were all significantly decreased compared with sham injury levels within the ipsilateral hippocampus after CCI. No significant molecular level changes were found in the contralateral hippocampus. Decreased expression of BDNF and synapsin I was also found within the ipsilateral parietal cortex of CCI‐injured rats compared with shams. However, BDNF and synapsin I expressions were significantly increased in the contralateral parietal cortex of the CCI rats. CREB expression was significantly decreased within the contralateral cortex of the CCI group. These findings show enduring reductions in the expression of BDNF, synapsin I, CREB, and α‐CAMKII ipsilateral to a CCI injury, which seem associated with the spatial learning deficits observed in this injury model. In addition, the delayed increase in the expression of BDNF and synapsin I within the cortex contralateral to CCI may reflect restorative processes in areas homotypical to the injury. © 2008 Wiley‐Liss, Inc.
Deletion of the amino‐terminal domain of the prion protein does not impair prion protein–dependent neuronal differentiation and neuritogenesis
Authors: Barenco Maria Grazia , Valori Chiara F. , Roncoroni Chiara , Loewer Johannes , Montrasio Fabio , Rossi Daniela
Published: October 2008

The cellular prion protein (PrPC) is a highly conserved glycoprotein of unknown biological function. To gain insight into the physiological role of PrPC, we generated a novel PrP knockout cell line, named PrPo/o ML, by immortalization of neuroepithelial precursor cells derived from the cerebellum of PrP‐knockout mice using the temperature‐sensitive simian virus 40 (SV40) large T antigen. We demonstrated that the PrPo/o ML cell line is a unipotent precursor line with glutamatergic properties, which can acquire neuronal features when cultivated under specific conditions. The role of the prion protein in the process of neuronal differentiation was then analyzed in the PrPo/o ML cells reconstituted with either the full‐length or an amino‐terminally deleted form of the prion protein. We show that the expression of PrPC facilitates the processes of neuronal differentiation and neuritogenesis and that the deletion of its amino‐terminal domain reduces the efficiency, but does not suppress this activity. This cell line represents a useful tool for studying PrP‐dependent signal transduction pathways during differentiation of neuronal stem/precursor cells. © 2008 Wiley‐Liss, Inc.
Regulation of relaxin 3 gene expression via cAMP‐PKA in a neuroblastoma cell line
Authors: Tanaka Masaki , Watanabe Yoshihisa , Yoshimoto Kanji
Published: October 2008

Relaxin 3 is expressed in neurons of the brain stem that inneravate wide areas of the forebrain. Relaxin 3 mRNA levels in these neurons are increased in response to restraint stress, and by central administration of corticotropin‐releasing factor (CRF). In the present study, we observed that relaxin 3 was expressed in a mouse neuroblastoma cell line, Neuro2a, and investigated the intracellular signaling that activated relaxin 3 gene transcription in vitro. By means of a clone stably transfected with a relaxin 3 promoter‐EGFP gene, we observed that dibutyryl cyclic AMP and forskolin increased the relaxin 3 promoter activity. These increases were inhibited by pretreatment with PKA inhibitors, H89 and KT5720. Moreover, the promoter activity was enhanced by CRF treatment after expression of CRF‐R1 receptor on the cells. Taken together, these results indicate that relaxin 3 transcription is activated via the cAMP‐PKA pathway in the downstream of CRF‐R1. © 2008 Wiley‐Liss, Inc.
Intracellular glutathione mediates the denitrosylation of protein nitrosothiols in the rat spinal cord
Authors: Romero Jorge M. , Bizzozero Oscar A.
Published: October 2008

Protein S‐nitrosothiols (PrSNOs) have been implicated in the pathophysiology of neuroinflammatory and neurodegenerative disorders. Although the metabolically instability of PrSNOs is well known, there is little understanding of the factors involved in the cleavage of S‐NO linkage in intact cells. To address this issue, we conducted chase experiments in spinal cord slices incubated with S‐nitrosoglutathione (GSNO). The results show that removal of GSNO leads to a rapid disappearance of PrSNOs (t½ ∼ 2 hr), which is greatly accelerated when glutathione (GSH) levels are raised with the permeable analogue GSH ethyl ester. Moreover, PrSNOs are stable in the presence of the GSH depletor diethyl maleate, indicating that GSH is critical for protein denitrosylation. Inhibition of GSH‐dependent enzymes (glutathione S‐transferase, glutathione peroxidase, and glutaredoxin) and enzymes that could mediate denitrosylation (alcohol dehydrogense‐III, thioredoxin and protein disulfide isomerase) do not alter the rate of PrSNO decomposition. These findings and the lack of protein glutathionylation during the chase indicate that most proteins are denitrosylated via rapid transnitrosylation with GSH. The differences in the denitrosylation rate of individual proteins suggest the existence of additional structural factors in this process. This study is relevant to our recent discovery that PrSNOs accumulate in the central nervous system of patients with multiple sclerosis. © 2008 Wiley‐Liss, Inc.
Effect of enzymes, flavor and organic acids on nutrient digestibility, performance and carcass traits of growing–finishing pigs fed diets containing dehydrated lucerne meal
Authors: Thacker Philip A , Haq Inam
Published: October 2008

BACKGROUND: We recently determined that 150 g kg−1 of lucerne meal can be fed during the finishing period without detrimental effects on pig performance but lucerne should be limited to less than 75 g kg−1 during the growing period. Lucerne has a high fiber content, is poorly digested and contains antinutritional factors such as saponins which reduce palatability. Therefore, pig performance could be enhanced by supplementing lucerne‐containing diets with feed additives such as flavors, enzymes or organic acids. The objective of this study was to determine the effects of these additives on nutrient digestibility, performance and carcass traits when fed to growing‐finishing swine fed diets containing lucerne.

RESULTS: Flavor and enzyme had no effect on nutrient digestibility, pig performance or carcass traits. During the growing period, inclusion of propionic and acetic acids increased (P < 0.05) the digestibility of dry matter, protein and energy. Weight gain and intake also increased (P < 0.05). None of the additives had any effect on pig performance during the finishing period or on carcass traits.

CONCLUSION: During the growing period, including 200 g kg−1 lucerne was detrimental to pig performance. Addition of organic acid improved pig performance and nutrient digestibility. During the finishing period, 200 g kg−1 lucerne can be successfully incorporated into pig diets without negative effects on pig performance. Copyright © 2008 Society of Chemical Industry
Rapid and simple method for screening of natural antioxidants from Chinese herb Flos Lonicerae Japonicae by DPPH‐HPLC‐DAD‐TOF/MS
Authors: Tang Dan , Li Hui‐Jun , Chen Jun , Guo Chao‐Wei , Li Ping
Published: October 2008

A rapid and simple method has been developed for the screening and identification of natural antioxidants of Flos Lonicerae Japonicae (FLJ), derived from the flower buds of Lonicera japonica. The hypothesis is that upon reaction with 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH), the peak areas (PAs) of compounds with potential antioxidant effects in the HPLC chromatograms will be significantly reduced or disappeared, and the identity confirmation could be achieved by HPLC‐DAD‐TOF/MS hyphenated technique. Using the proposed approach, about 14 compounds in the FLJ extract were found to possess a potential antioxidant activity. They were identified as chlorogenic acid (1), 1‐O‐caffeoylquinic acid (1‐O‐CQA, 2), caffeic acid (4), 4‐O‐CQA (5), rutin (7), isoquercitrin (8), luteolin‐7‐O‐glucoside (9), lonicerin (10), 4,5‐O‐dicaffeoylquinic acid (4,5‐O‐diCQA, 11), 3,5‐O‐diCQA (12), 1,3‐O‐diCQA (13), 3,4‐O‐diCQA (14), 1,4‐O‐diCQA (16), and luteolin (17). In addition, the free radical scavenging capacities of the available identified compounds were also investigated by HPLC assay. The results indicated that the compounds with PAs significantly decreasing were natural antioxidants, whereas those with PAs not changing presented no activities, which accordingly indicated that this newly proposed method could be widely applied for rapid screening and identification of natural antioxidants from complex matrices including Chinese herbal medicines.
Removal of envelope protein‐free retroviral vectors by anion‐exchange chromatography to improve product quality
Authors: Rodrigues Teresa , Alves Ana , Lopes António , Carrondo Manuel J. T. , Alves Paula M. , Cruz Pedro E.
Published: October 2008

We have investigated the role of the retroviral lipid bilayer and envelope proteins in the adsorption of retroviral vectors (RVs) to a FractogelTM DEAE matrix. Intact RVs and their degradation components (envelope protein‐free vectors and solubilized vector components) were adsorbed to this matrix and eluted using a linear gradient. Envelope protein‐free RVs (Env–) and soluble envelope proteins (gp70) eluted in a significantly lower range of conductivities than intact RVs (Env+) (13.7–30 mS/cm for Env– and gp70 proteins vs. 47–80 mS/cm for Env+). The zeta (ζ)‐potential of Env+ and Env– vectors was evaluated showing that envelope proteins define the pI of the viral particles (pI (Env+) < 2 versus 3 < pI (Env–) < 4) and that Env+ and Env– vectors have similar ζ‐potentials within pH 5 and 8. The results presented herein indicate that the adsorption of retroviral particles occurs through multi‐point interaction of the envelope proteins with the cationic groups on the chromatographic matrix. The strength of this adsorption is thus dependent on the amount of envelope protein present in the viral lipid bilayer. In conclusion, AEXc enables the separation of gp70 proteins as well as envelope protein‐free vectors constituting a significant improvement to the quality of retroviral preparations for gene therapy applications.
Analysis of volatiles in dehydrated carrot samples by solid‐phase microextraction followed by GC‐MS
Authors: Soria Ana Cristina , Sanz Jesús , Villamiel Mar
Published: October 2008

A solid‐phase microextraction (SPME)‐based method for the GC‐MS analysis of volatiles in dehydrated carrot root samples has been developed and the effect of the most important factors (fibre coating, extraction temperature, equilibrium time and extraction time) on the fractionation of different volatiles has been studied. GC‐MS chromatograms showed terpenoids relevant to carrot aroma such as α‐pinene, sabinene, β‐myrcene, limonene, γ‐terpinene, terpinolene, trans‐caryophyllene and β‐bisabolene, and several furan derivatives whose origin is discussed in this paper. As an example of application, this methodology has been used for the characterization of volatile composition of industrially dehydrated carrots. SPME followed by GC‐MS is shown as an affordable, fast and solvent‐free technique which can be performed with low sample amounts and be easily implemented at an industry for quality control purposes.
Determination of ascorbic acid in human plasma with a view to stability using HPLC with UV detection
Authors: Kand'ár Roman , Žáková Pavla
Published: October 2008

A method for the measurement of ascorbic acid using HPLC with UV detection and investigation into the protein precipitation techniques with regard to stability and recovery are described. The effectiveness of various protein precipitants was tested. Stability of ascorbic acid samples for analysis was investigated over 10 h. Ascorbic acid samples extracted with metaphosphoric acid were stable on a cooled autosampler (4°C) for at least 10 h (with a decline of 1.8% for ascorbic acid solution and 2.8% for plasma). Perchloric acid as protein precipitant for ascorbic acid was unsuitable (with a decline of 36.0% for ascorbic acid solution and 7.3% for plasma). Analytical performance of this method is satisfactory. The intra‐ and interassay coefficients of variation were 2.1% (n= 10) and 5.8% (n= 12), respectively. The calibration curve was linear with the tested range of 2.0–250.0 μmol/L. The recovery was 96.1% with CV = 4.8% (n= 6) and the LOD was 3 μmol/L. The preliminary reference ranges of ascorbic acid in a group of blood donors are 50.8 ± 22.4 μmol/L. This assay is a highly sensitive and reproducible HPLC method for the determination of ascorbic acid in human plasma.
Functionalized‐multiwalled carbon nanotubes as a preconcentrating probe for rapid monitoring of cationic dyestuffs in environmental water using AP‐MALDI/MS
Authors: Shrivas Kamlesh , Wu Hui‐Fen
Published: October 2008

A simple and sensitive method was developed for the rapid analysis of cationic dyestuffs from river and industrial wastewater using functionalized‐multiwalled carbon nanotubes (f‐MWCNT) with atmospheric pressure‐matrix assisted laser desorption/ionization mass spectrometry (AP‐MALDI/MS). The separation and preconcentration of analytes from sample solution was based on electrostatic force of attraction between positive dyestuffs and negatively charged f‐MWCNT. The optimum enrichment of the three dyestuffs was observed at pH 5.0 for 3 min contact time and using 1 mg f‐MWCNT in 1 mL water sample. The developed method has been successfully applied for the determination of three cationic dyestuffs namely neutral red (NR), brilliant cresyl blue (BB), and methylene blue (MB) in real world samples including river and industrial wastewater. The relative recoveries of dyestuffs from water sample were in the range 88.6–98.4%, indicating that the matrix had little effect on enrichment of analytes. The LOD and LOQ for cationic dyestuffs in water were 0.5–1.9 and 1.6–6.0 μg/L, respectively. All the results indicated that the proposed method could be used for the simultaneous determination of the three cationic dyestuffs in river and industrial wastewater at trace levels without the need of any chromatographic separation techniques.
Analysis of 51 persistent organic pollutants in soil by means of ultrasonic solvent extraction and stir bar sorptive extraction GC‐MS
Authors: Martínez‐Parreño Marta , Llorca‐Pórcel Julio , Valor Ignacio
Published: October 2008

A novel method based on ultrasonic solvent extraction and stir bar sorptive extraction (SBSE) for the analysis of 51 persistent organic pollutants including organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and polybrominated diphenylethers (PBDEs) in soil samples was developed. The different parameters that affect both the extraction of analytes from the soil samples, such as solvent selection, solvent volume, mass of soil, and extraction time, and the partitioning from the solvent/water mixture to the PDMS were studied. The final selected conditions consisted of the extraction of 1 g of soil with 15 mL methanol by sonication for 30 min. The methanol extract was mixed with 85 mL of Milli‐Q water and extracted by means of SBSE for 14 h at 900 rpm. The stir bars were analyzed by thermal desorption‐GC‐mass spectometry (TD‐GC‐MS). The effects of the matrix on the recovery of the various pollutants under the developed method were studied using two soils with very different physicochemical properties. Method sensitivity, linearity, repeatability, and reproducibility were also studied. Validation and accuracy of the method were conducted by analyzing two commercial certified reference materials (CRMs). The main advantage of this method resides in the fact that a small amount of a nontoxic solvent (methanol) is needed for the extraction of only 1 g of solid sample allowing LODs ranging from 0.01 to 2.0 μg/kg. Repeatability and reproducibility variations were lower than 20% for all investigated compounds. Results of the CRMs verify the high accuracy of this method.
Application of SPE‐HPLC‐DAD and SPE‐TLC‐DAD to the determination of pesticides in real water samples
Authors: Tuzimski Tomasz
Published: October 2008

Planar chromatography with diode array scanning (TLC‐DAD) and high‐performance chromatography with diode array detection (HPLC‐DAD) were used to screen water samples for pesticides. Pesticides were enriched from lake water samples by SPE on C18/SDB‐1, C18, C18 Polar Plus and cyanopropyl (CN) cartridges. The recovery rates were high for all extraction materials except for all pesticides on CN cartridges, for which the values were lower. SPE was used not only for preconcentration of analytes but also for their fractionation. The analytes were eluted first with methanol and then with dichloromethane. Methanol eluates were analysed by HPLC‐DAD, the dichloromethane eluates with TLC‐DAD. The method was validated for precision, repeatability and accuracy. The calibration plots were linear between 0.1 and 50.0 μg/mL for all pesticides, the correlation coefficients, r, were between 0.9992 and 1.000 as determined by HPLC‐DAD. In the TLC experiments, the best fit for the calibration lines was found when the calibration data were analysed using a second‐degree polynomial regression. Calibration plots lay between 0.1 and 17 μg/spot for all pesticides, the correlation coefficients, r, were between 0.9974 and 0.9997 determined by TLC‐DAD. The LOD was between 0.04 and 0.65 μg/spot (TLC‐DAD) and between 0.02 and 3.68 μg/mL (HPLC‐DAD).