bluuebunny/crossref_metadata_2025 huggingface dataset:
Transplants of Encapsulated Rat Choroid Plexus Cells Exert Neuroprotection in a Rodent Model of Huntington's Disease
Authors: Borlongan Cesario V. , Thanos Christopher G. , Skinner Steven J. M. , Geaney Marilyn , Emerich Dwaine F.
Published: April 2008

Choroid plexus (CP) epithelial cells secrete several neurotrophic factors and have been used in transplantation studies designed to impart neuroprotection against central nervous system (CNS) trauma. In the present study, CP was isolated from adult rats, encapsulated within alginate microcapsules, and transplanted unilaterally into the rat striatum. Three days later, unilateral injections of quinolinic acid (QA; 225 nmol) were made into the ipsilateral striatum to mimic the pathology observed in Huntington's disease (HD). After surgery, animals were tested for motor function using the placement test. Rats receiving CP transplants were significantly less impaired on this test. Nissl-stained sections demonstrated that CP transplants significantly reduced the volume of the striatal lesion produced by QA. Quantitative analysis of striatal neurons further demonstrated that choline acetyltransferase-immunoreactive, but not diaphorase-positive, neurons were protected by CP transplants. These data demonstrate that transplanted CP cells can be used to protect striatal neurons from excitotoxic damage and that the pattern of neuroprotection varies across specific neuronal populations.
Intravenously Administered Bone Marrow Cells Migrate to Damaged Brain Tissue and Improve Neural Function in Ischemic Rats
Authors: Wu Jiang , Sun Zhuo , Sun Hong-Shuo , Wu Jun , Weisel Richard D. , Keating Armand , Li Zhi-Hong , Feng Zhong-Ping , Li Ren-Ke
Published: April 2008

Accumulated evidence suggests that bone marrow stromal cells (BMSCs) are capable of regenerating damaged tissue. This study evaluated whether intravenously (noninvasively) administered, GFP-labeled BMSCs would migrate into damaged brain tissue and improve neurological function after a stroke. Wistar rats were subjected to middle cerebral artery occlusion and reperfusion. Twenty-four hours after injury, the rats received an IV injection of culture medium or BMSCs isolated from adult Wistar rats expressing green fluorescent protein (GFP). Two hours after injury and 1, 3, and 7 days after cell transplantation, neurological function was evaluated using a neurological severity scale. On day 7, the brain scar size was determined using tetrazolium chloride staining, and the implanted cells were identified using confocal microscopy. Immunohistochemistry was used to evaluate apoptosis and angiogenesis in the ischemic region, as well as the spatial distribution of the implanted BMSCs relative to the native neural cells. Implanted BMSCs migrated throughout the territory of the middle cerebral artery by 7 days after transplantation. Most implanted cells were located in the scar area and border zone of the ischemic region, and some expressed the neuronal marker NeuN. Rats receiving BMSC transplantation exhibited reduced scar size, limited apoptosis, and enhanced angiogenic factor expression and vascular density in the ischemic region relative to the control group, as well as significant improvements in the neurological severity scores. Intravenously administrated BMSCs facilitated the structural and functional recovery of neural tissue following ischemic injury, perhaps mediated by enhanced angiogenesis.
Cell Surface Heparan Sulfate Proteoglycans Mediate the Internalization of PDX-1 Protein
Authors: Ueda Michiko , Matsumoto Shinichi , Hayashi Shuji , Kobayashi Naoya , Noguchi Hirofumi
Published: April 2008

Although islet transplantation is a promising therapeutic option for the treatment of type 1 diabetes, the shortage of suitable donor tissues remains a major obstacle. Pancreatic stem/progenitor cells residing within the ductal epithelium have been used to generate human islet-like clusters, but there is no efficient strategy for facilitating differentiation of progenitor cells into insulin-producing cells. A previous study reported that exogenous PDX-1 protein can be transduced into pancreatic stem/progenitor cells and induce differentiation of the cells into insulin-producing cells without requiring gene transfer technology. This study provides genetic and biochemical evidence that cell membrane heparan sulfate proteoglycans are required for extracellular PDX-1 internalization. Heparin, one of the soluble glycosaminoglycans (GAGs), inhibited PDX-1 internalization, while chondroitin sulfate A, B, and C caused only very limited inhibition. Cell treatment with heparinase-III demonstrated impaired PDX-1 internalization, while treatment with chondroitinase ABC, or with chondroitinase AC, was completely ineffective in inhibiting PDX-1 internalization. Different mutant cell lines originating from CHO K1 cells and defective in GAG biosynthesis were also examined. PDX-1 internalization was significantly reduced in both pgs A-745 mutant cells, which are defective in a enzyme that initiates GAG synthesis, and pgs B-618 cells, which produce about 15% of the amount of GAGs synthesized by wild-type cells. These data indicate that cell-surface heparan sulfate proteoglycans are required for PDX-1 internalization and that PDX-1 protein transduction could be a valuable strategy for inducing insulin expression in pancreatic stem/progenitor cells without requiring gene transfer technology.
Cryopreservation of Mouse Adipose Tissue-Derived Stem/Progenitor Cells
Authors: Oishi Koichi , Noguchi Hirofumi , Yukawa Hiroshi , Miyazaki Takamichi , Kato Ryuji , Kitagawa Yasuo , Ueda Minoru , Hayashi Shuji
Published: April 2008

Adipose tissue-derived stem/progenitor cells (ASCs) have been reported to differentiate not only into mesodermal cells such as osteoblasts, chondorocytes, and adipocytes, but also to endodermal cells such as hepatocytes and insulin-expressing cells. These stem/progenitor cells are expected to be used for variety of regenerative therapies. This study demonstrates the viability and the adipo/osteogenic potential of cryopreserved ASCs using seven cryopreservation solutions, including 10% DMSO, Cell Freezing Medium-DMSO, Cell Freezing Medium-Glycerol, Cell Banker 1, Cell Banker 1+, Cell Banker 2, and CP-1. ASCs were obtained from mouse subcutaneous adipose tissue. The viability of the cryopreserved ASCs was over 90% with Cell Banker 2 preservation, approximately 90% with Cell Banker 1, Cell Banker 1+, or CP-1 preservation, and less than 80% for 10% DMSO, Cell Freezing Medium-DMSO, or Cell Freezing Medium-Glycerol preservation. No difference in the adipo/osteogenic potential was found between cells with or without cryopreservation in Cell Banker 2. These data suggests that Cell Banker 2 is the most effective cryopreservation solution for ASCs and that cryopreserved as well as noncryopreserved ASCs could be applied for regenerative medicine.
Secretory Unit of Islet in Transplantation (SUIT) and Engrafted Islet Rate (EIR) Indexes Are Useful for Evaluating Single Islet Transplantation
Authors: Noguchi Hirofumi , Yamada Yuichiro , Okitsu Teru , Iwanaga Yasuhiro , Nagata Hideo , Kobayashi Naoya , Hayashi Shuji , Matsumoto Shinichi
Published: April 2008

The evaluation of engraftment is important to assess the success of islet transplantation, but it is complex because islet transplantation usually requires two or more donors to achieve euglycemia. Islet transplantation from NHBDs was evaluated using new assessment forms for the secretory unit of islet in transplantation (SUIT) and engrafted islet rate (EIR) indexes. Insulin independence was obtained when the SUIT index was more than 28, which might indicate that 28% of the β-cell mass of a normal subject is required for insulin independence. Because the average EIR for a single transplantation is about 30, the percentage of engrafted islets following one transplantation is about 30%, assuming that a normal subject has 1 million islet equivalents. Although few cultured islet transplants have been performed, the increase of the SUIT and EIR indexes in patients who received cultured islets was significantly lower than in patients who received fresh islets, suggesting that fresh islets may be more effective than cultured islets. The SUIT and EIR indexes are thus considered to be useful values for evaluating islet transplantation, especially for single islet transplantation.
Heterotopic Transplant of an Isolated Rat Heart Preserved for 72 h in Perfluorocarbon with CO2
Authors: Yoshida Yu , Hatayama Naoyuki , Sekino Hajime , Seki Kunihiro
Published: April 2008

The inert fluid perfluorocarbon (PFC) has been used since about 1960 in liquid respiration and artificial blood for mammals. PFC has been used to successfully resuscitate tardigrades that had been dried and exposed to a high barometric pressure of 6,000 atmospheres. Next, scientists attempted to experimentally preserve organs that had been removed from animals, dried, and immersed in PFC. Since 1998 preservation and resuscitation experiments have been conducted with mammalian hearts using 2,015 rats and 70 pigs. Among those experiments, the maximum time after desiccation until successful resuscitation was 26 days for a rat heart and 37 days for a pig heart. However, these results could not be reproduced. Finally, in 2005, this laboratory demonstrated that a rat heart removed under 2 atmospheres pressure and a CO2 partial pressure of 400 hPa, followed by desiccation for 24 h, could be revived and heterotypically transplanted. Moreover, these results were reproducible. The preservation time can be extended to 72 h if, after immersing isolated rat hearts in PFC, they are dried by air exposure under a CO2 partial pressure of 100 hPa. The present report documents the resuscitation of this heart after 72 h of preservation followed by heterotrophic transplantation.
Bcl-2 Protects Tubular Epithelial Cells from Ischemia Reperfusion Injury by Inhibiting Apoptosis
Authors: Suzuki Chigure , Isaka Yoshitaka , Shimizu Shigeomi , Tsujimoto Yoshihide , Takabatake Yoshitsugu , Ito Takahito , Takahara Shiro , Imai Enyu
Published: April 2008

Ischemia followed by reperfusion leads to severe organ injury and dysfunction. Inflammation is considered to be the most important cause of graft dysfunction in kidney transplantation subjected to ischemia. The mechanism that triggers inflammation and renal injury after ischemia remains to be elucidated; however, cellular stress may induce apoptosis during the first hours and days after transplantation, which might play a crucial role in early graft dysfunction. Bcl-2 is known to inhibit apoptosis induced by the etiological factors promoting ischemia and reperfusion injury. Accordingly, we hypothesized that an augmentation of the antiapoptotic factor Bcl-2 may thus protect tubular epithelial cells by inhibiting apoptosis, thereby ameliorating the subsequent tubulointerstitial injury. We examined the effects of Bcl-2 overexpression on ischemia-reperfusion (I/R) injury using Bcl-2 transgenic mice (Bcl-2 TG) and their wild-type littermates (WT). To investigate the effects of I/R injury, the left renal artery and vein were clamped for 45 min, followed by reperfusion for 0–96 h. Bcl-2 TG exhibited decreased active caspase protein in the tubular cells, which led to a reduction in TUNEL-positive apoptotic cells. Consequently, interstitial fibrosis and phenotypic changes were ameliorated in Bcl-2 TG. In conclusion, Bcl-2 augmentation protected renal tubular epithelial cells from I/R, and subsequent interstitial injury by inhibiting tubular apoptosis.
Ductal Injection of Preservation Solution Increases Islet Yields in Islet Isolation and Improves Islet Graft Function
Authors: Noguchi Hirofumi , Ueda Michiko , Hayashi Shuji , Kobayashi Naoya , Okitsu Teru , Iwanaga Yasuhiro , Nagata Hideo , Nakai Yusuke , Matsumoto Shinichi
Published: April 2008

For islet transplantation, it is important to obtain an available islet mass adequate for diabetes reversal from a single donor pancreas. A recent report demonstrated that the use of M-Kyoto solution instead of UW solution improved islet yields in the two-layer method for pancreas preservation. The present study investigated whether the ductal injection of a large volume of preservation solution (UW and M-Kyoto solution) before pancreas storage improves islet yields. Islet yield both before and after purification was significantly higher in the ductal injection (+) group compared with the ductal injection (–) group. TUNEL-positive cells in the ductal injection (+) group were significantly decreased in comparison to the ductal injection (–) group. The ductal injection of preservation solution increased the ATP level in the pancreas tissue and reduced trypsin activity during the digestion step. Annexin V and PI assays showed that the ductal injection prevents islet apoptosis. In a transplant model, the ductal injection improved islet graft function. These findings suggest that the ductal injection of preservation solution, especially the M-Kyoto solution, leads to improved outcomes for pancreatic islet transplantation. Based on these data, this technique is now used for clinical islet transplantation from non-heart-beating donor pancreata or living donor pancreas.
Recombinant Sendai Virus-Mediated Gene Transfer to Adipose Tissue-Derived Stem Cells (ASCs)
Authors: Yukawa Hiroshi , Noguchi Hirofumi , Oishi Koichi , Miyazaki Takamichi , Kitagawa Yasuo , Inoue Makoto , Hasegawa Mamoru , Hayashi Shuji
Published: April 2008

Adipose tissue-derived stem cells (ASCs) are expected to have clinical applications as well as other stem cells, because ASCs can be obtained safely from adult donors and used in autologous therapies without concern about rejection and the need for immunosuppression. However, the use of gene transfer with Sendai virus (SeV) vectors, which can efficiently introduce foreign genes without toxicity into several cells, with ASCs has not yet been investigated. This study documents on the use of SeV vectors for gene transfer to ASCs. The dose-dependent GFP expression of ASCs transfected with SeV vectors after 48 h of culture at 37°C was first evaluated. Next, the cellular toxicity of ASCs transfected with SeV vectors was verified. In addition, SeV vectors were compared with adenovirus (AdV) vectors. Finally, the time-dependent GFP expression of ASCs transfected with SeV vectors was evaluated. The results showed that transfection of ASCs with SeV vectors results in more efficient expression of transgene (GFP expression) in the ASCs than with AdV vectors after 48 h of culture at 37°C. Moreover, while the transfection of ASCs with AdV vectors at high MOIs was cytotoxic (a lot of transfected cells died) that of ASCs with SeV vectors at high MOIs was not necessarily cytotoxic. In addition, the preservation of multilineage ASCs transfected with SeV was observed. In conclusion, this is the first report describing the successful use of SeV-mediated gene transfer in ASCs, and the results indicate that SeV may thus provide advantages with respect to safety issues in gene therapy.
Cloning and Characterization of Liver Progenitor Cells from the Scattered Cell Clusters in Primary Culture of Porcine Livers
Authors: Tokiwa Takayoshi , Yamazaki Taisuke , Ono Masashi , Enosawa Shin , Tsukiyama Takashi
Published: April 2008

The scattered cell clusters that can differentiate into hepatocytes or biliary epithelial cells have been isolated from primary cultures of adult porcine livers. We have generated 11 clonal cell lines from this system and identified liver progenitor cells (LPCs) among the clonal lines. These clonal lines expressed c-kit, HNF-1, HNF-6, and/or CK19 mRNA. An immunocytochemical study of the clonal lines indicated that clonal line CL-11 expressed liver epithelial cell markers CK14, vimentin, CK18, and BD-1. The expression of albumin and α1-antitrypsin (α1-AT) mRNA was only upregulated in CL-11 among the clonal lines when they were grown as aggregates. Under these conditions, CL-11 also exhibited ammonia metabolic activity and several indicators that suggest hepatocytic differentiation, including the upregulation of liver-specific genes such as dipeptidyl peptidase IV, CYP1A1, and CYP3A4 mRNA, and the downregulation of biliary cell markers such as γ-glutamyltrans-peptidase (GGT), CK19, and HNF6 mRNA. After culturing CL-11 in Matrigel, the expression of GGT and HNF6 mRNA was upregulated. These results indicate that CL-11 has dual potential: the ability to differentiate as hepatocytes or as bile duct cells. The isolation of scattered cells could provide a simple method to generate LPC lines from adult livers.
Serum Neutrophil Gelatinase-Associated Lipocalin as a Predictor of Organ Recovery from Delayed Graft Function after Kidney Transplantation from Donors after Cardiac Death
Authors: Kusaka Mamoru , Kuroyanagi Yoko , Mori Terumi , Nagaoka Kayuri , Sasaki Hitomi , Maruyama Takahiro , Hayakawa Kunihiro , Shiroki Ryoichi , Kurahashi Hiroki , Hoshinaga Kiyotaka
Published: April 2008

Because of a worldwide shortage of renal grafts, kidneys procured from donors after cardiac death (DCD) have recently become an important source of renal transplants. However, DCD kidneys often have complications with delayed graft function (DGF) and recipients require hemodialysis (HD) in the early period after kidney transplantation (KTx). This study evaluated serum NGAL as a potential specific parameter to predict early functional recovery of transplanted DCD kidneys. The average serum neutrophil gelatinase-associated lipocalin (NGAL) level in normal samples was 53 ± 30 ng/ml, while that in patients with chronic renal failure requiring HD was markedly raised at 963 ± 33 ng/ml. In patients undergoing a living-related KTx from a living donor (n = 11), serum NGAL level decreased rapidly after KTx, and only in two cases, with serum NGAL levels over 400 ng/ml on postoperative day 1 (POD1), was HD required due to DGF. In contrast, all patients undergoing a KTx from a DCD (n = 5) required HD due to DGF. Even in these cases, serum NGAL levels decreased rapidly several days after a KTx prior to the recovery of urine output and preceding the decrease in serum creatinine level. The pattern of decline in serum NGAL was biphasic, the decrease after the second peak indicating a functional recovery within the next several days. These data suggest that monitoring of serum NGAL levels may allow us to predict graft recovery and the need for HD after a KTx from a DCD.
Reestablishment of Microenvironment is Necessary to Maintain In Vitro and In Vivo Human Islet Function
Authors: Navarro-Alvarez Nalú , Rivas-Carrillo Jorge David , Soto-Gutierrez Alejandro , Yuasa Takeshi , Okitsu Teru , Noguchi Hirofumi , Matsumoto Shinichi , Takei Jiro , Tanaka Noriaki , Kobayashi Naoya
Published: April 2008

Islet transplantation is associated with an elevated rate of early graft failure. The isolation process leads to structural and functional abnormalities. The reestablishment of the cell–matrix relationship is important to modulate the survival and function of islets. Thus, we evaluated the effect of human fibronectin (hFN) and self-assembling peptide nanofiber (SAPNF) in the ability to support islet function in vitro and after transplantation into streptozotocin (STZ)-induced diabetic severe combined immunodeficiency (SCID) mice. Human isolated islets were cultured with hFN or SAPNF for 7 days. Their ability to maintain insulin production/glucose responsiveness over time was evaluated. Islets embedded in hFN, SAPNF, or alone were transplanted into STZ-induced diabetic SCID mice. Islet grafts were removed after 14 days to evaluate insulin content, insulin expression, and apoptosis. SAPNF-entrapped islets maintained satisfactory morphology/viability and capability of glucose-dependent insulin secretion for over 7 days, whereas islets cultured in hFN underwent widespread deterioration. In vivo grafts containing human islets in SAPNF showed remarkably higher insulin content and expression when compared with human islets in hFn or alone. RT-PCR revealed lower caspase-3 expression in SAPNF islets grafts. These studies indicate that the reestablishment of the cell–matrix interactions by a synthetic matrix in the immediate postisolation period is a useful tool to maintain islet functions in vitro and in vivo.
Binge drinking and violence against intimate partners in Russia
Authors: Lysova Aleksandra V. , Hines Denise A.
Published: April 2008

This study is the first to provide information on the association between intimate partner violence (IPV) and binge drinking among Russian university students. Using data from 500 (58% female) university students from the four Russian sites of the International Dating Violence Study, we found gender differences in rates of IPV perpetration and in the association between binge drinking and IPV. Specifically, more females than males perpetrated IPV, and the associations between binge drinking and IPV were stronger for the female students than for the male students. In addition, antisocial traits and behavior (ATSB) were significantly related to both binge drinking and IPV perpetration for males and females. For males, the relatively weak associations between binge drinking and IPV perpetration disappeared once ASTB was accounted for. For females, the relationship decreased but remained significant when ATSB was statistically controlled. Path analyses confirmed that this pattern of relationships would be consistent with ATSB serving as a partial mediator between binge drinking and IPV perpetration. However, other alternative mediation and moderation models for the relationships between binge drinking, IPV perpetration, and ATSB could not be ruled out with this one‐wave correlational study. Aggr. Behav. 34:416–427, 2008. © 2008 Wiley‐Liss, Inc.
Unrelated chromosomal anomalies found in patients with suspected 22q11.2 deletion
Authors: Fernández Luis , Lapunzina Pablo , López Pajares Isidora , Palomares María , Martínez Isabel , Fernández Blanca , Quero José , García‐Guereta Luis , García‐Alix Alfredo , Burgueros Margarita , Galán‐Gómez Enrique , Carbonell‐Pérez José María , Pérez‐Granero Ángeles , Torres‐Juan Laura , Heine‐Suñer Damià , Rosell Jordi , Delicado Alicia
Published: April 2008

Screening for 22q11.2 deletions has not an easy approach due to the wide variability of their associated phenotype. Many clinical features overlap with those of other known syndromes and reported loci. Patients referred to exclude a 22q11.2 deletion are usually tested with a locus‐specific FISH probe, with 10% positive cases depending on the selection criteria, but patients testing negative for FISH at 22q11.2 may have other chromosomal aberrations in routine cytogenetic analysis. We tested 819 patients suspected of having a 22q11.2 deletion. Eighty‐eight patients (10.7%) were positive for 22q11.2 deletion, whereas 30 patients (3.7%) showed other chromosomal abnormalities involving deletions and duplications, derivative chromosomes, marker chromosomes, apparently balanced and unbalanced translocations and sex chromosome aneuploidies. Of these alterations, 28 did not involve region 22q11 and most had not been associated with 22q11.2 deletion phenotype before. We discuss the similarity of DiGeorge/velocardiofacial syndrome with other known clinical entities and suggest correlations between the new loci and the observed clinical features. The frequency of unrelated chromosomal anomalies reported in this study and in other previous reports highlights the importance of conventional cytogenetic analysis as an initial genome‐wide screening tool in all referred patients, and provides useful data to optimize diagnostic and screening protocols according to the most frequent chromosomal findings. © 2008 Wiley‐Liss, Inc.
A new candidate locus for bilateral perisylvian polymicrogyria mapped on chromosome Xq27
Authors: Santos Neide F. , Secolin Rodrigo , Brandão‐Almeida Iara L. , Silva Marilza S. , Torres Fábio R. , Tsuneda Simone S. , Guimarães Catarina A. , Hage Simone R.V. , Cendes Fernando , Guerreiro Marilisa M. , Lopes‐Cendes Iscia
Published: April 2008

Polymicrogyria (PMG) is characterized by an excessive number of small and prominent brain gyri, separated by shallow sulci. Bilateral perisylvian polymicrogyria (BPP) is the most common form of PMG. Clinical signs include pseudobulbar paresis, mental retardation, and epilepsy. Familial forms of BPP have been described and a candidate locus was previously mapped to chromosome Xq28, distal do marker DXS8103. The objective of this study was to perform linkage analysis in one family segregating BPP. A total of 15 individuals, including 8 affected patients with BPP were evaluated. Family members were examined by a neurologist and subjected to magnetic resonance imaging scans. Individuals were genotyped for 18 microsatellite markers, flanking a 42.3 cM interval on ch Xq27‐q28. Two‐point and multipoint linkage analysis was performed using the LINKAGE package and haplotype reconstruction was performed by GENEHUNTER software. Our results showed a wide spectrum of clinical manifestations in affected individuals with BPP, ranging from normal to mild neurological abnormalities. Two‐point linkage analysis yield a Zmax = 2.06 at θ = 0.00 for markers DXS1205 and DXS1227. Multipoint lod‐scores indicate a candidate interval of 13 cM between markers DSXS1205 and DXS8043, on ch Xq27.2‐Xq27.3. These results point to a new locus for BPP in a more centromeric location than previously reported. © 2008 Wiley‐Liss, Inc.
Association study of candidate variants of COMT with neuroticism, anxiety and depression
Authors: Wray Naomi R. , James Michael R. , Dumenil Troy , Handoko Herlina Y. , Lind Penelope A. , Montgomery Grant W. , Martin Nicholas G.
Published: April 2008

The Val158Met polymorphism of the gene encoding catechol‐O‐methyltransferase (COMT) is one of the most widely tested variants for association with psychiatric disorders, but replication has been inconsistent including both sex limitation and heterogeneity of the associated allele. In this study we investigate the association between three SNPs from COMT and anxiety and depression disorders and neuroticism all measured within the same study sample. Participants were selected as sibling pairs (or multiples) that were either concordant or discordant for extreme neuroticism scores from a total sample of 18,742 Australian twin individuals and their siblings. All participants completed the Composite International Diagnostic Interview (CIDI) from which diagnoses of DSM‐IV depression and anxiety disorders were determined. Of the participants, 674 had a diagnosis of anxiety and/or depression from 492 families. Study participants (n = 2,045 from 987 families) plus, where possible, their parents were genotyped for rs737865, rs4680 (Val158Met), and rs165599. Using family based tests we looked for association between these variants and neuroticism, depression, anxiety, panic disorder and agarophobia (PDAG) and obsessive compulsive disorder. We found no convincing evidence for association either in allelic or genotypic tests for the total sample or when the sample was stratified by sex. Haplotype T‐G‐G showed weak association (P = 0.042) with PDAG before correction for multiple testing; association between this haplotype and schizophrenia has been previously reported in an Australian sample. © 2008 Wiley‐Liss, Inc.
Combined effects of exonic polymorphisms in CRHR1 and AVPR1B genes in a case/control study for panic disorder
Authors: Keck Martin E. , Kern Nikola , Erhardt Angelika , Unschuld Paul G. , Ising Marcus , Salyakina Daria , Müller Marianne B. , Knorr Carolin C. , Lieb Roselind , Hohoff Christa , Krakowitzky Petra , Maier Wolfgang , Bandelow Borwin , Fritze Jürgen , Deckert Jürgen , Holsboer Florian , Müller‐Myhsok Bertram , Binder Elisabeth B.
Published: April 2008

Accumulating evidence from animal studies suggests that the corticotropin releasing hormone (CRH) and arginine vasopressin (AVP) neuropeptide systems, contribute to anxiety behavior. To investigate whether polymorphisms in the genes regulating these two systems may alter susceptibility to anxiety disorders in humans, we genotyped 71 single nucleotide polymorphisms (SNPs) in CRH, CRHR1, CRHR2, AVP, AVPR1A, AVPR1B in a German sample from Munich with patients suffering from panic disorder and matched healthy controls (n = 186/n = 299). Significant associations were then replicated in a second German sample with 173 patients with panic disorder and 495 controls. In both samples separately and the combined sample, SNPs within CHRH1 and AVPR1B were nominally associated with panic disorder. We then tested two locus multiplicative and interaction effects of polymorphisms of these two genes on panic disorder. Fifteen SNP pairs showed significant multiplicative effects in both samples. The SNP pair with the most significant association in the combined sample (P = 0.00057), which withstood correction for multiple testing, was rs878886 in CRHR1 and rs28632197 in AVPR1B. Both SNPs are of potential functional relevance as rs878886 is located in the 3′ untranslated region of the CRHR1 and rs28632197 leads to an arginine to histidine amino acid exchange at position 364 of AVPR1B which is located in the intracellular C‐terminal domain of the receptor. These data suggest that polymorphisms in the AVPR1B and the CRHR1 genes alter the susceptibility to panic disorder. © 2008 Wiley‐Liss, Inc.
Social support in older individuals: The role of the BDNF Val66Met polymorphism
Authors: Taylor Warren D. , Züchner Stephan , McQuoid Douglas R. , Steffens David C. , Blazer Dan G. , Krishnan K. Ranga R.
Published: April 2008

Although often viewed as a purely environmental construct, perception of social support may be influenced by genetic factors. This study examined the relationship between the brain‐derived neurotrophic factor (BDNF) Val66Met polymorphism and social support measures in older subjects. The sample consisted of 243 depressed and 115 nondepressed older subjects, age 60 years or older; 233 were Val66 allele homozygotes, while 125 were Met66 allele carriers. All subjects completed clinical assessments, including a self‐report questionnaire assessing four social support domains, and provided blood for genotyping. Statistical models examined the relationship between scale scores of social support and BDNF Val66Met genotype, while controlling for presence or absence of major depressive disorder and other demographic factors significantly associated with social support. As social support measures were not normally distributed, log‐transformed scores were examined. After controlling for diagnosis and education level, the Met66 allele was associated with lower levels of subjective social support (F1,357 = 5.33, P = 0.0216) and a trend for fewer social interactions (F1,357 = 3.66, P = 0.0567). To our knowledge, this is the first report associating a measure of social support with a genetic polymorphism. This supports previous work that genetic factors may influence social support perception. Further work is needed to determine the generalizability of this finding to the broader population, as well as its significance for clinical outcomes. © 2008 Wiley‐Liss, Inc.
A de novo apparently balanced translocation [46,XY,t(2;9)(p13;p24)] interrupting RAB11FIP5 identifies a potential candidate gene for autism spectrum disorder
Authors: Roohi Jasmin , Tegay David H. , Pomeroy John C. , Burkett Sandra , Stone Gary , Stanyon Roscoe , Hatchwell Eli
Published: April 2008

Autism spectrum disorder (ASD) is a severe developmental disorder of the central nervous system characterized by impairments in social interaction, communication, and range of interests and behaviors. The syndrome's prevalence is estimated to be as high as 1 in 150 American children yet its etiology remains largely unknown. Examination of observed cytogenetic variants in individuals with ASD may identify genes involved in its pathogenesis. As part of a multidisciplinary study, an apparently balanced de novo translocation between chromosomes 2 and 9 [46,XY,t(2;9)(p13;p24)] was identified in a subject with pervasive developmental disorder not otherwise specified (PDD‐NOS), and no distinctive dysmorphic features. Molecular characterization of the rearrangement revealed direct interruption of the RAB11 family interacting protein 5 (RAB11FIP5) gene. RAB11FIP5 is a Rab effector involved in protein trafficking from apical recycling endosomes to the apical plasma membrane. It is ubiquitously expressed and reported to contribute to both neurotransmitter release and neurotransmitter uptake at the synaptic junction. Detailed analysis of the rearrangement breakpoints suggests that the reciprocal translocation may have formed secondary to incorrect repair of double strand breaks (DSBs) by nonhomologous end‐joining (NHEJ). © 2008 Wiley‐Liss, Inc.
Enzymatic ring‐opening polymerization of ε‐caprolactone by a new lipase from Yarrowia lipolytica
Authors: Barrera‐Rivera Karla A. , Flores‐Carreón Arturo , Martínez‐Richa Antonio
Published: April 2008

A new lipase from the yeast Yarrowia lipolytica was isolated and used in the enzymatic ring‐opening polymerization of lactones. The effect of used commercial oil from a vacuum pump (instead of olive oil) and in the presence of wheat flour was evaluated. It was found that lipase production is favored when wheat flour and used commercial oil is added instead of olive oil with an incubation time of 16.5 h at 29°C and 250 rpm. Lipase shows a specific activity of 3.47 nmoles of 4‐methyl umbelliferone released/mg of protein/min. In this way, preculture step can be avoided and an important reduction in production time can be obtained. In vitro polymerization of ε‐caprolactone at different temperatures in the presence of n‐heptane for 15 days produces low‐molecular‐weight polyesters with unique multiphase morphology. Polyesters were characterized by NMR (solution and solid‐state) spectroscopy, differential scanning calorimetry (DSC), Fourier‐transformed infrared, wide‐angle X‐ray scattering and MALDI‐TOF. Polyester crystallinities calculated by DSC were high, as expected for low‐molecular‐weight PCLs. Final polymer possesses carboxylic acid and hydroxyl end‐groups, as determined by 1H‐ and 13C NMR analysis. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci 2008
Tracking anthropogenic‐ and climatic‐related environmental changes in the remaining habitat lakes of the endangered Atlantic whitefish (Coregonus huntsmani) using palaeolimnological techniques
Authors: Ginn Brian K. , Grace Laura C. , Cumming Brian F. , Smol John P.
Published: April 2008

Atlantic whitefish (Coregonus huntsmani) is a critically endangered species whose remaining habitat is restricted to three lakes near Bridgewater, Nova Scotia, Canada. Other lakes in this region have been affected by a variety of environmental changes over the past 150 years (e.g. acidic deposition, eutrophication, and climatic changes); however, the extent of the impact on these remaining Atlantic whitefish lakes, and how the current limnological conditions compare with pre‐industrial conditions, is not known.

Given the lack of long‐term monitoring data, palaeolimnological techniques were used to track environmental changes in these three lakes to infer historic limnological conditions.

Results of this study show that acidic deposition has had no significant impact on these lakes (diatom‐inferred lakewater pH has changed little over time), nor has the nutrient status of these lakes changed as the sediment profiles have been consistently dominated by oligotrophic diatom taxa.

Changes in the dominance of diatom assemblages since ∼1850, fromAulacoseira distanstoCyclotella stelligera, are correlated with climatic warming (r = 0.48–0.89,P<0.05).

Contrary to initial concerns, these only remaining habitats of the Atlantic whitefish have not yet been affected by acidic precipitation. However, other potential stressors, such as climatic warming and associated limnological changes, may now be affecting the habitat of this endangered species.

Copyright © 2008 John Wiley & Sons, Ltd.
Regarding Several Points of Doubt of the Structure of the Olfactory Bulb: As Described by T. Blanes
Authors: Levine Catherine , Marcillo Alexander
Published: April 2008

In order to complement the studies on the fila olfactoria completed by Dr. Santiago Ramón y Cajal, we have included a translation of “Sobre Algunos Puntos Dudosos de la Estructura del Bulbo Olfatorio” by T. Blanes, a student of Dr. Cajal. This work describes in stunning detail additional morphological aspects of the olfactory pathway, including what was at the time the modestly studied neuroglia. The neuroglia of the olfactory system has been revisited in the last several decades for its importance in the field of regenerative neuroscience. Olfactory ensheathing glia has the unique quality of providing ensheathment to neurons which traverse from the central to the peripheral nervous system and are being used as a candidate in present‐day transplantation studies to mimic this phenomenon at the dorsal root entry zone after a central nervous system injury. Although this fine work has passed its centennial anniversary since initial publication, it has been widely cited throughout the years, and of recent when Pressler and Stowbridge reported Blanes cell electrophysiological recordings (Neuron V 49, 6; p 889–904, 2006). An English translation the details of what Blanes initially documented with unduplicated precision can now be made available to a wider audience in the field of neuroscience, and is especially important now that more and more present‐day studies require a precise and complete understanding of the anatomical structures contained within the olfactory system. Anat Rec, 291:751‐762, 2008. © 2008 Wiley‐Liss, Inc.
Van der Waals and Polar Intermolecular Contact Distances: Quantifying Supramolecular Synthons
Authors: Ganguly Parthasarathy , Desiraju Gautam R.
Published: April 2008

Crystal structures are viewed as being determined by ranges and constraints on interatomic contact distances between neighboring molecules. These distances are considered to arise from environment‐dependent atomic sizes, that is, larger sizes for isotropic, van der Waals type contacts and smaller sizes for more‐polar, possibly ionic contacts. Although the idea of different, or anisotropic, radii for atoms is not new, we developed a method of obtaining atomic sizes that is based on a theoretical framework. Using different atomic sizes for the same atom in different environments, we were able to rationalize some structural observations and anomalies. For example, benzene with the Pbca structure may be described in terms of two types of C⋅⋅⋅H interactions: a longer contact largely of the van der Waals type, and a shorter, structure‐determining type (Cδ−⋅⋅⋅Hδ+), which we term “n‐polar”. Our approach is illustrated with three examples: 1) the equivalence in crystal packing of fluorobenzene, benzonitrile, pyridine N‐oxide, and pyridine/HF 1:1 molecular complex, all of which take the not‐so‐common tetragonal P41212 space group and are practically isomorphous; 2) the similarity of the Pa3 acetylene and Pbca benzene crystal structures; and 3) the equivalence between an increase in pressure and an increase in the “n‐polar” contacts in Pbca benzene; in other words, the equivalence between hydrostatic pressure and chemical pressure. In the context of crystal engineering, we describe a method whereby the topological information conveyed in a supramolecular synthon is recast in a more quantitative manner. A particular synthon, and in turn the crystal structure to which it leads, is viable within small ranges of distances of its constituent atoms, and these distances are determined by chemical factors.
Comparative analysis on microbial and rat metabolism of ginsenoside Rb1 by high‐performance liquid chromatography coupled with tandem mass spectrometry
Authors: Chen Guangtong , Yang Min , Song Yan , Lu Zhiqiang , Zhang Jinqiang , Huang Huilian , Guan Shuhong , Wu Lijun , Guo De‐an
Published: April 2008

Ginsenoside Rb1 is an active protopanaxadiol saponin from Panax species. In order to compare the similarities and differences of microbial and mammalian metabolisms of ginsenoside Rb1, the microbial transformation by Acremonium strictum and metabolism in rats were comparatively studied. Microbial transformation of ginsenoside Rb1 by Acremonium strictum AS 3.2058 resulted in the formation of eight metabolites. Ten metabolites (M1–M10) were detected from the in vivo study in rats and eight of them were identified as the same compounds as those obtained from microbial metabolism by liquid chromatography–tandem mass spectrometry analysis and comparison with reference standards obtained from microbial metabolism. Their structures were identified as ginsenoside Rd, gypenoside XVII, 20(S)‐ginsenoside Rg3, 20(R)‐ginsenoside Rg3, ginsenoside F2, compound K, 12β‐hydroxydammar‐3‐one‐20(S)‐O‐β‐d‐glucopyranoside, and 25‐hydroxyl‐(E)‐20(22)‐ene‐ginsenoside Rg3, respectively. The structures of the additional two metabolites were tentatively characterized as 20(22),24‐diene‐ginsenoside Rg3 and 25‐hydroxyginsenoside Rd by HPLC‐MS/MS analysis. M7–M10 are the first four reported metabolites in vivo. The time course of rat metabolism of ginsenoside Rb1 was also investigated. Copyright © 2008 John Wiley & Sons, Ltd.
Establishment of a HPLC method for preclinical pharmacokinetic study of the novel anti‐Parkinson's disease candidate drug FLZ in rats
Authors: Li Li‐bo , Zhang Jin‐lan , Wang Yu‐xiang , Wei Huai‐ling , Liu Geng‐tao
Published: April 2008

An HPLC method was established and validated for the determination of compound FLZ, a synthetic novel anti‐Parkinson's disease candidate drug, in rat plasma. FLZ and the internal standard bicyclol were extracted from plasma by solid‐phase extraction method and analyzed on a Restek C18 column (4.6 × 250 mm, 5 µm) with a mobile phase consisting of methanol and water (60:40, v/v) at a flow rate of 1.0 mL/min. The detection wavelength was set at 320 nm. The calibration curve was linear within the concentration range from 25 to 500 ng/mL (r2 > 0.999), the limit of quantitation was 25 ng/mL and the average recovery was 92.0% with the RSD less than 5.9%. The relative standard deviation for intra‐ and inter‐day precision was less than 3.8 and 6.9%, respectively. The established HPLC method was validated to be a simple, rapid and reliable procedure and applied to study the preclinical pharmacokinetics of FLZ in rat plasma, and it was the first time that the pharmacokinetics of FLZ had been investigated. Copyright © 2008 John Wiley & Sons, Ltd.
Environmental motivations: a classification scheme and its impact on environmental strategies and practices
Authors: Paulraj Antony
Published: April 2008

Ecological deterioration has significantly changed the role of businesses in society and the way they are perceived. In fact, consumers are more aware of environmental issues and are demanding that the businesses take appropriate action in preserving the environment. Extant research suggests that stringent regulations had been instrumental to the timing and direction of many firms' responses to environmental preservation. In addition to these legislative requirements, firms also adopt ecologically responsive practices for various other reasons. Therefore, understanding the different motivations of environmental initiatives is pertinent. Additionally, it is also important to determine whether environmental strategies and practices differ significantly across firms with different motivations. With the above ambition in mind, this manuscript addresses the crucial issues related to corporate environmental strategy through the development of an empirical taxonomy of environmental motivations and the subsequent testing of any significant differences in corporate environmental strategy and green practices across the clusters. Copyright © 2008 John Wiley & Sons, Ltd and ERP Environment.
Photolabile Glutamate Protecting Group with High One‐ and Two‐Photon Uncaging Efficiencies
Authors: Gug Sylvestre , Charon Sébastien , Specht Alexandre , Alarcon Karine , Ogden David , Zietz Burkhard , Léonard Jérémie , Haacke Stefan , Bolze Frédéric , Nicoud Jean‐François , Goeldner Maurice
Published: April 2008

A π‐extended [2‐(2‐nitrophenyl)propoxy]carbonyl (NPPOC) derivative has been prepared as an efficient UV and near‐IR photolabile protecting group for glutamate. This glutamate cage compound exhibits efficient photorelease upon one‐photon excitation (εΦ=990 M−1 cm−1 at 315 nm). In addition, it also shows efficient photorelease in activation of glutamate receptors in electrophysiological recordings. Combined with a high two‐photon uncaging cross‐section (δΦ=0.45 GM at 800 nm), its overall properties make this new cage—3‐(2‐propyl)‐4′‐methoxy‐4‐nitrobiphenyl (PMNB)—for glutamate a very promising tool for two‐photon neuronal studies.
Fluorescent Agonists for the Torpedo Nicotinic Acetylcholine Receptor
Authors: Krieger Florian , Mourot Alexandre , Araoz Romulo , Kotzyba‐Hibert Florence , Molgó Jordi , Bamberg Ernst , Goeldner Maurice
Published: April 2008

We have synthesized a series of fluorescent acylcholine derivatives carrying different linkers that vary in length and structure and connect the acylcholine unit to the environment‐sensitive fluorophores 7‐(diethylamino)coumarin‐3‐carbonyl (DEAC) or N‐(7‐nitrobenz‐2‐oxa‐1,3‐diazol‐yl) (NBD). The pharmacological properties of the fluorescent analogues were investigated on heterologously expressed nicotinic acetylcholine receptor (nAChR) from Torpedo californica and on oocytes transplanted with nAChR‐rich Torpedo marmorata membranes. Agonist action strongly depends on the length and the structure of the linker. One particular analogue, DEAC‐Gly‐C6‐choline, showed partial agonist behavior with about half of the maximum response of acetylcholine, which is at least 20 times higher than those observed with previously described fluorescent dansyl‐ and NBD‐acylcholine analogues. Binding of DEAC‐Gly‐C6‐choline to Torpedo nAChR induces a strong enhancement of fluorescence intensity. Association and displacement kinetic experiments revealed dissociation constants of 0.5 nM for the αδ‐binding site and 15.0 nM for the αγ‐binding site. Both the pharmacological and the spectroscopic properties of this agonist show great promise for characterizing the allosteric mechanism behind the function of the Torpedo nAChR, as well as for drug‐screening studies.
High‐Relaxivity Magnetic Resonance Imaging (MRI) Contrast Agent Based on Supramolecular Assembly between a Gadolinium Chelate, a Modified Dextran, and Poly‐β‐Cyclodextrin
Authors: Battistini Elisa , Gianolio Eliana , Gref Ruxandra , Couvreur Patrick , Fuzerova Silvia , Othman Mohammad , Aime Silvio , Badet Bernard , Durand Philippe
Published: April 2008

Nanosized contrast agents have great potential in magnetic resonance molecular imaging applications for clinical diagnosis. This study proposes new nanoparticles spontaneously formed under mild conditions and composed of a noncovalent adduct between a gadolinium complex, a polymer of β‐cyclodextrin (pβCD: MW 1.5×106 g mol−1) and a dextran grafted with alkyl chains (MD). The formation of this supramolecular nanoassembly is based upon a “lock‐and‐key” recognition process in which the hydrophobic alkyl chains of MD and the adamantyl moieties of macrocyclic GdIII chelates are included in the cavities of pβCD. The large number of βCDs contained in the pβCD resulted in the formation of 200 nm diameter nanoparticles, each entrapping 1.8×105 molecules of a low‐molecular‐weight Gd complex. This system, which exhibits a great relaxivity enhancement (48.4 mM−1 s−1, at 20 MHz and 37 °C) compared to the GdIII chelate itself (5.2 mM−1 s−1), appears to be a promising strategy for the in vivo targeted delivery of GdIII complexes. The mechanisms of particle formation, conjugation strategies, and relaxometric characterizations in the field of contrast‐enhanced magnetic resonance imaging are discussed.