bluuebunny/crossref_metadata_2025 huggingface dataset:
Spermatozoa modulate epididymal cell proliferation and protein secretion in vitro
Authors: Reyes‐Moreno Carlos , Laflamme Julie , Frenette Gilles , Sirard Marc‐André , Sullivan Robert
Published: September 2007

Normal epididymal function, such as protein expression and secretion, is primarily regulated by testicular androgens and temperature. However, the role of spermatozoa in this critical process has never been studied. In order to determine whether sperm itself could regulate epididymal function, we have developed a cell culture system of bovine epididymal cells to study the interactions between spermatozoa and the epididymal epithelium. Primary cells from caput, corpus, and cauda epididymal tissues were cultured in the presence of androgens at 32°C (scrotal) and 37°C (abdominal). Newly synthesized proteins were metabolically labeled with35S‐methionine after sperm co‐incubation and the pattern of secreted proteins was analyzed by two‐dimensional polyacrylamide gel electrophoresis. Proliferation rate, protein secretion rate and electrophoretic patterns of secreted proteins were evaluated 48 hr post‐co‐incubation. Incubation at 32°C indicated that spermatozoa stimulation increases the level of protein secretion of cultured cells from all epididymal sections while it slightly decreases proliferation of corpus cells. At 37°C, spermatozoa co‐incubation significantly decreases the protein secretion rate of cultured cells from all epididymal sections. Independently of cell incubation temperature, spermatozoa stimulation induces both an increase in the intensity of radiolabeled proteins and the appearance of new secreted proteins of caput cells without affecting the protein pattern of corpus or cauda cells. Incubation at 37°C, however, greatly modifies the pattern of proteins expressed at 32°C by cauda cells. Taken together, these results support the hypothesis that spermatozoa themselves affect epididymal cell function, most importantly for caput epididymides. Mol. Reprod. Dev. 75: 512–520, 2008. © 2007 Wiley‐Liss, Inc.
Cumulus cells accelerate aging of mouse oocytes by secreting a soluble factor(s)
Authors: Qiao Tian‐Wu , Liu Na , Miao De‐Qiang , Zhang Xia , Han Dong , Ge Li , Tan Jing‐He
Published: September 2007

Control of oocyte aging in vitro is important for both human‐assisted reproduction and animal embryo technologies because fertilization or artificial activation of aged oocytes results in abnormal development. Interactions between somatic and germ cells are also an important issue in current biological research. The role of cumulus cells (CCs) in maturation, ovulation, and fertilization of oocytes has been extensively studied, yet little is known about their role in oocyte aging. Although our previous study has shown that CCs accelerate the aging progression of mouse oocytes, the mechanism by which CCs accelerate oocyte aging is unknown. In this study, cumulus‐denuded mouse oocytes (DOs) were co‐cultured with cumulus‐oocyte complexes (COCs) or CC monolayer or cultured in medium conditioned with these cells and changes in the susceptibility to activating stimuli and in MPF activity of oocytes were evaluated after different aging treatments. The results showed that culture with or in medium conditioned with COCs or CC monolayer promoted activation of DOs, indicating that a soluble factor is responsible for the aging‐promoting effect. The in vivo and in vitro‐matured DOs did not differ in responsiveness to the aging‐promoting factor (APF). Heat shock did not accelerate oocyte aging unless in the presence of CCs. The production of APF was not affected by the age or maturation system of COCs, but increased with their density and duration of culture. The results strongly suggest that CCs accelerated oocyte aging by secreting a soluble APF into the medium. Further analysis showed that the APF was heat labile but stable to freezing, it had a threshold effective concentration and can be depleted by DOs. Mol. Reprod. Dev. 75: 521–528, 2008. © 2007 Wiley‐Liss, Inc.
Vitrification of in vitro matured ovine oocytes affects in vitro pre‐implantation development and mRNA abundance
Authors: Succu Sara , Bebbere Daniela , Bogliolo Luisa , Ariu Federica , Fois Stefano , Leoni Giovanni Giuseppe , Berlinguer Fiammetta , Naitana Salvatore , Ledda Sergio
Published: September 2007

The impact of vitrification procedures on in vitro matured (IVM) ovine oocytes mRNA content and ability to undergo successful fertilization, cleavage and embronic development was assessed. Vitrified‐warmed (n = 113) and control (n = 140) IVM oocytes were in vitro fertilized and cultured up to blastocyst stage under standard conditions. Vitrified oocytes showed lower cleavage rate (47% vs. 75%, P < 0.001) and development to blastocyst stage (17% vs. 57%, P < 0.001) than controls. In addition, the timings of the first cleavage and blastocysts production were significantly delayed in the vitrified‐warmed group (P < 0.001 in both cases). In parallel, we analyzed by reverse transcriptase real‐time PCR the relative abundance of β‐actin, H2A.Z histone, Poli A Polimerase (PAP), Heat Shock Protein 90β (HSP90β), P34cdc2, Cyclin b, Na/K‐ATPase and Type I cadherin (E‐Cad) transcripts in single IVM controls (n = 24) and vitrified‐warmed oocytes (n = 40). Results were normalized against the exogenous rabbit α‐globin mRNA standard and the β‐actin housekeeping gene and similarly described a lower abundance of most mRNAs in oocytes subjected to vitrification procedures. When normalized against the exogenous standard mRNA, all transcripts except for β‐actin and H2A.Z showed a significantly different abundance in the two classes of oocytes. The same results were obtained after normalization against the internal standard, except for HSP90β and E‐Cad transcripts, whose lower abundance in vitrified‐warmed oocytes resulted prominent, but not significant (P = 0.083 and P = 0.068, respectively). The oocyte lower transcripts abundance following vitrification might be an early indicator of poor quality in good correlation with the developmental data to blastocyst stage. Mol. Reprod. Dev. 75: 538–546, 2008. © 2007 Wiley‐Liss, Inc.
Role of nonessential amino acids on porcine embryos produced by parthenogenesis or somatic cell nuclear transfer
Authors: Gupta Mukesh Kumar , Uhm Sang Jun , Lee Sung‐ho , Lee Hoon Taek
Published: September 2007

Amino acids play a multitude of roles during early embryonic development and have been demonstrated to facilitate improved development of in vivo or in vitro fertilized and parthenogenetic embryos in several species. However, review of emerging literatures, shows that culture milieu of cloned embryos might be different from those of in vitro fertilized embryos. This study therefore, evaluated the effect of nonessential amino acids (NEAA) on yield and quality of porcine embryos produced by somatic cell nuclear transfer (SCNT) and compared them with parthenogenetic embryos as control. Analysis showed that, supplementation of NEAA to culture medium significantly improved the blastocyst rate of parthenogenetic (38.9 ± 8.8 vs. 27.5 ± 9.0%) as well as SCNT (22.5 ± 2.2 vs. 13.8 ± 3.4%) embryos although cleavage rates were not different (P < 0.05). These blastocysts also had higher hatching ability and contained higher cell number than control blastocysts (P < 0.05). TUNEL labeling revealed that blastocysts cultured in the presence of NEAA were less predisposed to biochemical apoptosis and showed lower indices of TUNEL, fragmentation, and total apoptosis than those cultured in the absence of NEAA (P < 0.05). Real‐time qRT‐PCR analysis further revealed that NEAA decreased the expression ratio of BAX:BCL‐xL and enhanced the relative abundance of IGF2 transcripts. Therefore, our study suggests that NEAA improves the yield and quality of cloned porcine embryos by enhancing blastocyst expansion, hatching, and total cell number and decreasing the apoptosis by positively modulating the expression of embryo survival related genes, similar to those reported for in vivo or in vitro fertilized embryos. Nonessential amino acids improve the yield and quality of cloned and parthenogenetic porcine embryos and modulate the expression of embryo survival related genes. Mol. Reprod. Dev. 75: 588–597, 2008. © 2007 Wiley‐Liss, Inc.
Properties of murine embryonic stem cells maintained on human foreskin fibroblasts without LIF
Authors: Meng G.L. , zur Nieden N.I. , Liu S.Y. , Cormier J.T. , Kallos M.S. , Rancourt D.E.
Published: September 2007

In embryonic stem (ES) cells, leukemia inhibitory factor (LIF)/STAT3, wnt and nodal/activin signaling are mainly active to control pluripotency during expansion. To maintain pluripotency, ES cells are typically cultured on feeder cells of varying origins. Murine ES cells are commonly cultured on murine embryonic fibroblasts (MEFs), which senesce early and must be frequently prepared. This process is laborious and leads to batch variation presenting a challenge for high‐throughput ES cell expansion. Although some cell lines can be sustained by exogenous LIF, this method is costly. We present here a novel and inexpensive culture method for expanding murine ES cells on human foreskin fibroblast (HFF) feeders. After 20 passages on HFFs without LIF, ES cell lines showed normal expression levels of pluripotency markers, maintained a normal karyotype and retained the ability to contribute to the germline. As HFFs do not senesce for at least 62 passages, they present a vast supply of feeders. Mol. Reprod. Dev. 75: 614–622, 2008. © 2007 Wiley‐Liss, Inc.
Global gene expression analysis of bovine blastocysts produced by multiple methods
Authors: Zhou Wenli , Xiang Tianhao , Walker Shawn , Farrar Vicki , Hwang Earl , Findeisen Brian , Sadeghieh Sanaz , Arenivas Fernando , Abruzzese Ronald V. , Polejaeva Irina
Published: September 2007

Reproductive efficiency using somatic cell nuclear transfer (SCNT) technology remains suboptimal. Of the various efforts to improve the efficiency, chromatin transfer (CT) and clone‐clone aggregation (NTagg) have been reported to produce live cloned animals. To better understand the molecular mechanisms of somatic cell reprogramming during SCNT and assess the various SCNT methods on the molecular level, we performed gene expression analysis on bovine blastocysts produced via standard nuclear transfer (NT), CT, NTagg, in vitro fertilization (IVF), and artificial insemination (AI), as well as on somatic donor cells, using bovine genome arrays. The expression profiles of SCNT (NT, CT, NTagg) embryos were compared with IVF and AI embryos as well as donor cells. NT and CT embryos have indistinguishable gene expression patterns. In comparison to IVF or AI embryos, the number of differentially expressed genes in NTagg embryos is significantly higher than in NT and CT embryos. Genes that were differentially expressed between all the SCNT embryos and IVF or AI embryos are identified. Compared to AI embryos, more than half of the genes found deregulated between SCNT and AI embryos appear to be the result of in vitro culture alone. The results indicate that although SCNT methods have altered differentiated somatic nuclei gene expression to more closely resemble that of embryonic nuclei, combination of insufficient reprogramming and in vitro culture condition compromise the developmental potential of SCNT embryos. This is the first set of comprehensive data for analyzing the molecular impact of various nuclear transfer methods on bovine pre‐implantation embryos. Mol. Reprod. Dev. 75: 744–758, 2008. © 2007 Wiley‐Liss, Inc.
Alterations in the testis of hormone sensitive lipase‐deficient mice is associated with decreased sperm counts, sperm motility, and fertility
Authors: Hermo Louis , Chung Shari , Gregory Mary , Smith Charles E. , Wang Shu Pei , El‐Alfy Mohamed , Cyr Daniel G. , Mitchell Grant A. , Trasler Jacquetta
Published: September 2007

Hormone‐sensitive lipase (HSL, Lipe, E.C.3.1.1.3) functions as a triglyceride and cholesteryl esterase, supplying fatty acids, and cholesterol to cells. Gene‐targeted HSL‐deficient (HSL−/−) mice reveal abnormal spermatids and are infertile at 24 weeks after birth. The purpose of this study was to follow the evolution of spermatid abnormalities as HSL−/− mice age, characterize sperm motility in older HSL−/− mice, and determine if mice expressing a human testicular HSL transgene (HSL−/−ttg) produce normal motile sperm. In situ hybridization indicated that HSL is expressed exclusively in steps 5–16 spermatids, but not in Sertoli cells. In HSL−/− mice, abnormalities were evident in step 16 spermatids at 5 weeks after birth, with defects progressively increasing in spermatids with age. The defects included multinucleation of spermatids, abnormal shapes and a reduction of elongating spermatids. In older HSL−/− mice, sperm counts appeared reduced by 42%, but this value was lower because samples were compromised by the presence of small degenerating germ cells in addition to sperm, both of which appeared of similar size and density. Sperm motility was dramatically reduced with only 11% classified as motile in HSL−/− mice compared to 76–78% of sperm in wild‐type and HSL−/−ttg mice. Sperm morphology, counts, and motility were normal in HSL−/−ttg mice, as was their fertility. Collectively, the data indicate that HSL deficiency results in abnormal spermatid development with defects arising at 5 weeks of age and progressively increasing at later ages. HSL−/− mice also show a dramatic reduction in sperm counts and motility and are infertile. Mol. Reprod. Dev. 75: 565–577, 2008. © 2007 Wiley‐Liss, Inc.
Enhancement of Bovine oocyte maturation by leptin is accompanied by an upregulation in mRNA expression of leptin receptor isoforms in cumulus cells
Authors: van Tol Helena T.A. , van Eerdenburg Frank J.C.M. , Colenbrander Ben , Roelen Bernard A.J.
Published: September 2007

In this study, the mechanisms of supposed leptin action on oocyte maturation were examined. Expression of leptin mRNA, as determined with RT‐PCR, was present in oocytes but not in cumulus cells. The long isoform of the leptin receptor (ObR‐L) was expressed exclusively in cumulus cells after 7 and 23 hr of maturation. In oocytes the expression of the short receptor isoform (ObR‐S) and all the receptor isoforms combined (ObR‐T) did not change during maturation, as determined by quantitative RT‐PCR, but in cumulus cells there was a significant increase in ObR‐S transcripts after 7 hr of maturation. To determine if leptin plays a role in resumption of meiosis, oocytes meiotically arrested by the connection of the cumulus to a piece of granulosa layer were exposed to leptin. After 23 hr of culture, the proportion of oocytes that had resumed meiosis did not differ from the control. Exposure of COCs to leptin (1,000 ng/ml) resulted after 17 hr of maturation in a smaller proportion of oocytes that was still in metaphase‐I stage (M‐I) compared to the control group. Fertilization of oocytes after maturation in the presence of leptin resulted in a larger proportion of embryos that had developed to the 8‐cell stage on Day 5 compared to the control group and in more blastocysts on Day 8 of culture. It is concluded that leptin enhances meiotic maturation of bovine oocytes, and that this effect is cumulus cell‐mediated. Mol. Reprod. Dev. 75: 578–587, 2008. © 2007 Wiley‐Liss, Inc.
Aberrant DNA methylation imprints in aborted bovine clones
Authors: Liu Jing‐He , Yin Shen , Xiong Bo , Hou Yi , Chen Da‐Yuan , Sun Qing‐Yuan
Published: September 2007

Genomic imprinting plays a very important role during development and its abnormality may heavily undermine the developmental potential of bovine embryos. Because of limited resources of the cow genome, bovine genomic imprinting, both in normal development and in somatic cell nuclear transfer (SCNT) cloning, is not well documented. DNA methylation is thought to be a major factor for the establishment of genomic imprinting. In our study, we determined the methylation status of differential methylated regions (DMRs) of four imprinted genes in four spontaneously aborted SCNT‐cloned fetuses (AF). Firstly, abnormal methylation imprints were observed in each individual to different extents. In particular, Peg3 and MAOA were either seriously demethylated or showed aberrant methylation patterns in four aborted clones we tested, but Xist and Peg10 exhibited relatively better maintained methylation status in AF1 and AF4. Secondly, two aborted fetuses, AF2 and AF3 exhibited severe aberrant methylation imprints of four imprinted genes. Finally, MAOA showed strong heterogeneous methylation patterns of its DMR in normal somatic adult tissue, but largely variable methylation levels and relatively homogeneous methylation patterns in aborted cloned fetuses. Our data indicate that the aborted cloned fetuses exhibited abnormal methylation imprints, to different extent, in aborted clones, which partially account for the higher abortion and developmental abnormalities during bovine cloning. Mol. Reprod. Dev. 75: 598–607, 2008. © 2007 Wiley‐Liss, Inc.
A novel approach to identify bovine sperm membrane proteins that interact with receptors on the vitelline membrane of bovine oocytes
Authors: Pate Barry J. , White Kenneth L. , Chen Dong , Aston Kenneth I. , Sessions Benjamin R. , Bunch Thomas D. , Weimer Bart C.
Published: September 2007

At fertilization, the sperm triggers intracellular calcium oscillations, which are pivotal to oocyte activation and development. A working hypothesis for the interaction between the sperm and the oocyte is that disintegrin ligands on the inner acrosomal membrane of the sperm bind to integrin receptors on the oocyte vitelline membrane. The aim of these experiments was to find and identify the sperm protein ligands involved in bovine sperm–oocyte interactions. In situ fluorescent labeling of proteins and 2‐D gel electrophoresis were used to identify specific sperm membrane proteins that interact with proteins in the oocyte vitelline membrane. Sperm were labeled with a fluorescent dye and used to fertilize zona‐free oocytes. Sperm–oocyte complexes were either lysed immediately, or following covalent cross‐linking of proteins with dibromobimane. The cross‐linking reagent serves the critical function of covalently linking proteins together so that they will remain as a unit through lysis of the cells and 2‐D gel analysis, and which can be subsequently identified by mass spectrometry. Lysates were electrophoretically run on the same 2‐D gel. The comparison of uncross‐linked and cross‐linked protein spots revealed that some proteins shifted position based on binding. These spots were picked and proteins identified by mass spectrometry. These results provide a list of specific sperm proteins that interact with oocyte membrane proteins and establish a group of candidate ligands, one or more of which may be responsible for induction of outside‐in signaling resulting in oocyte activation and fusion of the gametes. Mol. Reprod. Dev. 75: 641–649, 2008. © 2007 Wiley‐Liss, Inc.
Siloxane/silane‐crosslinked systems from supercritical carbon dioxide: I. Fluorinated poly(ether/siloxane)s
Authors: Zhou Hui , Venumbaka Sreenu R. , Fitch John W. , Cassidy Patrick E.
Published: September 2007

A series of new, fluorinated, silicone‐containing polymers with crosslinkable units has been synthesized by hydrosilation in toluene and supercritical carbon dioxide (70°C, 3000 psi) using platinum‐divinyltetramethyldisiloxane (Pt‐DVTMS) as a catalyst. The polymers were characterized by FTIR, NMR, GPC, TGA, and DSC. The molecular weights of these polymers ranged from 9,900 to 41,000. Comparison of the properties between reactions in toluene versus supercritical carbon dioxide indicated that the green solvent is a suitable alternative for hydrosilation. The hydrolysis and thermal curing of the crosslinkable precursor polymers produced higher molecular weight polymers with thermal stabilities ranging from 347 to 417°C. Copyright © 2007 John Wiley & Sons, Ltd.
Thermo and pH sensitive fluorescent polymer sensor for metal cations in aqueous solution
Authors: Liu Yunhai , Meng Lingzhi , Lu Xiaoju , Zhang Lifen , He Yongbing
Published: September 2007

A new monomeric fluorophore N‐acryl‐N'‐(quinolin‐8‐yl)thiourea (AQT) was synthesized. Free radical copolymerization was carried out in order to prepare a water‐soluble fluorescent copolymer, based on N‐isopropylacrylamide. The fluorescent characteristic of the aqueous solutions of copolymers was investigated both in varied pH and in the presence of metal cations. The polymer‐ contained AQT was found to be a selective chemosensor for Ni, Pb, Co ions especially for Co ion. The obtained copolymers show thermo‐sensitive lower critical solution temperature (LCST) ranging from 32.5 to 37.5°C with varied ratios of AQT, N, N‐dimethylacrylamide and N‐isopropylamide. Copyright © 2007 John Wiley & Sons, Ltd.
Influence of doped polyaniline on the interaction of Pu/PAni blends and on its microwave absorption properties
Authors: Biscaro Rogério S. , Rezende Mirabel C. , Faez Roselena
Published: September 2007

In this work, the influence of polyaniline (PAni) doped with both camphorsulfonic acid (PAni‐CSA) and dodecylbenzenesulfonic acid (PAni‐DBSA) on polyurethane (PU)/PAni blends was studied by rheological and morphological analyses. The effect of doped polyaniline on the attenuation of incident microwave radiation, in the frequency range from 8.0 to 12.0 GHz, was also investigated. The complex viscosity (η*) of PAni‐DBSA blends is observed to vary more significantly as a function of resting time than PAni‐CSA blends. This behavior is attributed to a better dispersion of PAni particles into the matrix on account of the presence of smaller agglomerates, as observed by optical and electron microscopy. However, this behavior has not been determinant on microwave absorption by the blends, with those that contain PAni‐CSA showing higher attenuation values. Copyright © 2007 John Wiley & Sons, Ltd.
Prognostic significance of immunohistochemical expression of ezrin in non‐metastatic high‐grade osteosarcoma
Authors: Ferrari Stefano , Zanella Licciana , Alberghini Marco , Palmerini Emanuela , Staals Eric , Bacchini Patrizia
Published: September 2007

Background

Ezrin is a membrane‐cytoskeleton linker protein involved in regulating growth and metastatic behaviour of cancer cells. The study evaluated ezrin expression and its prognostic value in patients with non‐metastatic osteosarcoma.

Procedures

Ezrin expression and pattern of staining (cytoplasmic or membraneous and cytoplasmic) were assessed using immunohistochemistry on slides from tumour biopsy. We studied 95 patients (median 16 years, range: 4–39 years) with primary non‐metastatic osteosarcoma of the extremity treated by neoadjuvant chemotherapy based on methotrexate, cisplatin, doxorubicin and ifosfamide.

Results

Seventy‐six patients (80%) showed ezrin immunoreactivity: in cytoplasm (37, 47%) and in cytoplasm and membrane (42, 53%) of tumour cells. Immunohistochemical staining score was: 1+ (16, 24%), 2+ (10, 13%), 3+ (17, 21.5%) and 4+ (36, 45.5%). Ezrin and score expression were not related to gender, site, alkaline phosphatase (AP), LDH serum levels, chemotherapy‐induced tumour necrosis or patient outcome. A significant association was seen between expression pattern and prognosis. The 3‐year probability of disease‐free survival was 80% for patients with only cytoplasmic immunostaining and 54% for patients with cytoplasmic and membranous immunostaining (P < 0.02).

Conclusion

Ezrin immunoreactivity can be detected in the majority of patients with non‐metastatic osteosarcoma of the extremity. The pattern of ezrin staining can identify patients with different risks of relapse. In patients who only have ezrin cytoplasmic expression, a probability of EFS >80% at 5 years can be expected. These results suggest further investigations to define the relation between expression pattern, ezrin functional status and outcome in patients with non‐metastatic osteosarcoma. Pediatr Blood Cancer 2008;50:752–756. © 2007 Wiley‐Liss, Inc.
Cordocentesis in multifetal pregnancies
Authors: Tongprasert Fuanglada , Tongsong Theera , Wanapirak Chanane , Sirichotiyakul Supatra , Piyamongkol Wirawit
Published: September 2007

Objective

To describe the experiences in diagnostic cordocentesis in twin pregnancies at midpregnancy

Methods

The database and medical records of pregnant women attending Maternal Fetal Medicine Unit of the hospital for diagnostic cordocentesis at midpregnancy between January 1989 and September 2006 were retrospectively reviewed.

Results

During 17 years of experience, 4241 cordocenteses at midpregnancy were performed for prenatal diagnosis, including 59 procedures in 30 multiple pregnancies (29 twins and 1 triplet). The mean gestational age at the time of cordocentesis was 19.5 ± 1.6 weeks. Success rate of the samplings was 98.3% with one sample was maternal blood contamination. Averaged‐time used of the procedures was 8.2 minutes (range 1–45 minutes). The procedure‐related complications included transient bleeding at puncture site (8.5%) and transient fetal bradycardia (22.0%). The total fetal loss rate was 10.5% but there was no cordocentesis‐related fetal loss (0.0%), defined as a fetal loss within 2 weeks after the procedure.

Conclusion

This study may provide a new insight on the safety of cordocentesis in multifetal pregnancies at midpregnancy. The procedure‐related fetal loss is not as high as reported in the past. This study suggests cordocentesis be a relatively safe and highly successful in obtaining fetal blood samples. Copyright © 2007 John Wiley & Sons, Ltd.
Predicting adverse neonatal outcome in severe fetal compromise: A pivotal role for estimated fetal weight Z scores
Authors: Moreira de Sa Renato Augusto , Salomon Laurent J. , Carvalho Paulo Roberto N. , Lopes Laudelino Marques , Ville Yves
Published: September 2007

Objective

To identify significant predictors for adverse neonatal outcome in severe by compromised fetuses.

Methods

Consecutive premature fetuses at between 25 and 32 weeks' with severe placental insufficiency were examined prospectively. Inclusion criteria were: singletons; normal anatomy; abnormal umbilical artery Doppler pulsatility index; abnormal cerebroplacental ratio; middle cerebral artery pulsatility index (PI) ⩽2 standard deviations. We considered as independent potential predicting variables: absent or reversed end diastolic flow in umbilical artery, abnormal ductus venosus S/A ratio, absent or reversed flow during atrial contraction in the ductus venosus, and estimated fetal weight (EFW) Z score. Outcome parameters were: neonatal mortality and severe neonatal morbidity (Periventricular‐Intraventricular Hemorrhage grade 03 or 04, Retinopathy of Prematurity stage 3 or 4, Periventricular Leukomalacia or Broncopulmonary Dysplasia).

Results

Univariate analysis for all potential predicting variables was performed. EFW Z score index showed the strongest association with mortality and there was no fetal death or Retinopathy of Prematurity in the group with EFW Z‐score index below lower limit. There was no case of Cystic Periventricular Leukomalatia in these analyses. The area under receiver‐operator characteristic curve was significant for EFW Z‐score index.

Conclusion

Our study suggests that EFW Z score is the strongest predictor of adverse neonatal outcome in severely compromised fetuses. Copyright © 2007 John Wiley & Sons, Ltd.
Prenatal diagnosis of xeroderma pigmentosum and trichothiodystrophy in 76 pregnancies at risk
Authors: Kleijer Wim J. , van der Sterre Marianne L. T. , Garritsen Victor H. , Raams Anja , Jaspers Nicolaas G. J.
Published: September 2007

Objective

Evaluation of results in a consecutive series of 76 prenatal diagnoses for xeroderma pigmentosum (XP) and trichothiodystrophy (TTD) made since 1977.

Methods

UV‐induced DNA repair synthesis was assessed by the autoradiographic measurement of the incorporation of 3H‐thymidine.

Results

XP was diagnosed in 19 of the 76 investigated pregnancies at risk; cultured chorionic villus (CV) cells were used in 33 pregnancies with ten affected fetuses and cultured amniocytes in 43 pregnancies with nine affected fetuses. In four cases, CVS results were corroborated by subsequent investigation of amniocytes because maternal cell contamination in the CV cell culture was either present or could not be excluded. Uncertain results in two other cases with intermediate DNA repair capacity and severe maternal cell contamination required further investigation. Median time needed for cell culture and analysis was 25 days. To reduce intra‐assay variations, a modification of the DNA repair synthesis assay has recently been developed. In this assay, patients and controls are investigated simultaneously in mixed cultures of cells labelled with polystyrene beads.

Conclusion

Reliable prenatal diagnosis for XP and TTD can be made by the demonstration of clearly reduced UV‐induced DNA repair synthesis due to defective global genome nucleotide excision repair. Copyright © 2007 John Wiley & Sons, Ltd.
Nanocomposites of poly(L‐lysine) and single‐walled carbon nanotubes
Authors: Kim Jong‐Beom , Premkumar Thathan , Giani Olivia , Robin Jean‐Jaques , Schue Francois , Geckeler Kurt E
Published: September 2007

BACKGROUND: Single‐walled carbon nanotubes have inspired research owing to their promise in a broad range of applications. The dispersion of carbon nanotubes is of key importance for the utilization of this interesting material for various potential applications.

RESULTS: A novel and simple method was developed to fabricate polymer composites with single‐walled carbon nanotubes based on a solid‐state reaction, in which the nanotubes were reacted with poly(L‐lysine) using high‐speed vibration milling. Fourier transform infrared and UV‐visible spectroscopy as well as thermogravimetry were employed to characterize the novel composites. The morphology and the dispersion of the carbon nanotubes were determined using scanning and transmission electron microscopy.

CONCLUSION: The resulting composites were dispersable in water and are expected to have great potential for both molecular‐level studies and device applications of nanotubes. Copyright © 2007 Society of Chemical Industry
Effective biosynthesis of poly(3‐hydoxy‐ butyrate‐co‐4‐hydroxybutyrate) with high 4‐hydroxybutyrate fractions by Wautersia eutropha in the presence of α‐amino acids
Authors: Kimura Hiroshi , Ohura Takeshi , Matsumoto Taisuke , Ikarashi Takuya
Published: September 2007

BACKGROUND: Biopolymers produced by microbes are in demand as their biodegradable and biocompatible properties make them suitable for disposable products and for potential use as biomaterials for medical applications. The effective microbial production of copolyesters of 3‐hydroxybutyrate (3HB) and 4‐hydroxybutyrate(4HB) with high molar fractions of 4HB unit by a wild‐type Wautersia eutropha H16 was investigated in culture media containing 4‐hydroxybutyric acid (4HBA) and different carbon substrates in the presence of various α‐amino acids.

RESULTS: The addition of carbon sources such as glucose, fructose and acetic acid to the culture medium containing 4HBA in the presence of α‐amino acids resulted in the production of random poly(3HB‐co‐4HB) with compositions of up to 77 mol% 4HB unit, but the yields of copolyesters with 60–77 mol% 4HB units were less than 15 wt% of dried cell weights. In contrast, when carbon sources such as propionic acid and butyric acid were used as the co‐substrates of 4HBA in the presence of α‐amino acids, poly(3HB‐co‐4HB) copolyesters with compositions of 72–86 mol% 4HB were produced at maximally 47.2 wt% of dried cell weight (11.3 g L−1) and the molar conversion yield of 4HBA to 4HB fraction in copolyesters was as high as 31.4 mol%. Further, poly(3HB‐co‐4HB) copolyesters with compositions of 93–96 mol% 4HB were isolated at up to 35.2 wt% of dried cell weights by fractionation of the above copolymers with chloroform/n‐hexane.

CONCLUSION: The productivity of copolyesters with over 80 mol% 4HB fractions was as high as 0.146 g L−1 h−1 (3.51 g L−1 for 24 h) by flask batch cultivation. Copyright © 2007 Society of Chemical Industry
Luminescence properties of poly‐ (phenylene vinylene) derivatives
Authors: Ton‐That Cuong , Stockton Geoff , Phillips Matthew R , Nguyen Thien‐Phap , Huang Chun Hao , Cojocaru Ala
Published: September 2007

BACKGROUND: Conjugated polymers, especially those of the poly(phenylene vinylene) (PPV) family, are promising candidates as emission material in light‐emitting devices. The aim of this work was to investigate the dependence of the luminescence properties of PPV‐based derivatives on their polymer structure, especially side groups.

RESULTS: Three PPV derivatives, BEHPPV, MEHPPV and MEHSPPV, were synthesised and characterised by photoluminescence (PL) and cathodoluminescence (CL) spectroscopies in the temperature range 10–300 K. PL and CL spectra of the polymers exhibit similar luminescence peaks, which undergo a blue shift with increasing temperature. The shift in wavelength is accompanied by variations in the relative intensities of emission peaks. Both BEHPPV and MEHPPV display emission characteristics of the PPV backbone, but the peak of MEHPPV shifts to a longer wavelength in comparison with the corresponding peak of BEHPPV at the same temperature. The luminescence spectra of MEHSPPV, which has a sulfanyl incorporated in the side chain, are considerably different from those of the two other derivatives.

CONCLUSIONS: The results demonstrate that the luminescence properties depend strongly on the chain conformations of the conjugated backbone, which are affected by polymer side chains. Copyright © 2007 Society of Chemical Industry
Hydration, entropy, and energy relationships for periodate oxidations
Authors: Taylor Jay E.
Published: September 2007

Periodate oxidations of ethanediol and pinacol each occur in two phases; these are (1) formation and (2) decomposition of the intermediate complex. In phase (1), an increase in acidity gives ${\rm IO}_4^{-} \to {\rm H}_5 {\rm IO}_6 \to {\rm H}_6 {\rm IO}_6^{+}$. The rate of oxidation of ethanediol decreases with increasing acidity, whereas the rate of oxidation of pinacol maximizes with H5IO6. For both glycols, the activation energy increases and ΔSact decreases with increasing acidity. In phase (2), the energy of activation is essentially constant with pH, whereas the rate decreases, and the entropy of activation decreases modestly as pH decreases. The latter correlates with the nonhomogeniuty of product formation. Rates for 3‐chloro‐1,2‐propanediol are also listed. Pentaerythritol forms an inactive complex with ${\rm IO}_4^{{-} }$ or H5IO6 indicating the importance of chelation in the formation of the intermediate complex. Copyright © 2007 John Wiley & Sons, Ltd.
How do women who choose not to participate in population‐based cervical cancer screening reason about their decision?
Authors: Blomberg Karin , Ternestedt Britt‐Marie , Törnberg Sven , Tishelman Carol
Published: September 2007

In Stockholm, Sweden, women are invited to a cost‐free population‐based cervical cancer screening programme (PCCSP) at regular intervals. Despite this, many women choose not to attend screening at all or to take opportunistic tests instead. This study explores how women who actively declined participation in the PCCSP reasoned about their choice. Qualitative telephone interviews and fax messages from women who actively declined participation in the PCCSP were analysed inductively. The manner in which women defined and conceptualized distinctions between, and the roles and responsibilities of, both private and public spheres were found to be central in explanations of decision making. Factors related to women's decisions not to participate in screening at all include a lack of confidence in the benefits of screening, previous negative health care and preventive experiences, a belief in one's own ability to discern health changes or a belief that one was not at risk for cervical cancer, as well as a number of unconventional standpoints on social and political issues. Women who chose not to participate in the organized PCCSP, but who did use private opportunistic screening, generally motivated this with direct or indirect criticism of the screening programme itself. Not only was the examination itself sensitive but also all facets of the PCCSP, from invitation letter on, were found to influence women's decisions. Using Jepson et al.'s ethical framework to peruse the evidence‐base underlying women's ‘informed decision‐making’ about CCS is suggested to be more constructive than discussing potential participants' knowledge versus lack of knowledge. Copyright © 2007 John Wiley & Sons, Ltd.
Automatic prediction of protein interactions with large scale motion
Authors: Schneidman‐Duhovny Dina , Nussinov Ruth , Wolfson Haim J.
Published: September 2007

Proteins often change their conformation upon binding to other molecules. Taking these conformational changes into account in docking is an extremely difficult task: the larger the scale of the motion the harder it is to predict the structure of the association complex. Here, we present a fully automated method for flexible docking with large scale motion in one of the docked molecules. The method automatically identifies hinge regions and rigid parts and then docks the input molecules while explicitly considering the hinges and possible protein motions. Proteins 2007. ©2007 Wiley‐Liss, Inc.
Altered pesticide use on transgenic crops and the associated general impact from an environmental perspective
Authors: Kleter Gijs A , Bhula Raj , Bodnaruk Kevin , Carazo Elizabeth , Felsot Allan S , Harris Caroline A , Katayama Arata , Kuiper Harry A , Racke Kenneth D , Rubin Baruch , Shevah Yehuda , Stephenson Gerald R , Tanaka Keiji , Unsworth John , Wauchope R Donald , Wong Sue‐Sun
Published: September 2007

The large‐scale commercial cultivation of transgenic crops has undergone a steady increase since their introduction 10 years ago. Most of these crops bear introduced traits that are of agronomic importance, such as herbicide or insect resistance. These traits are likely to impact upon the use of pesticides on these crops, as well as the pesticide market as a whole. Organizations like USDA‐ERS and NCFAP monitor the changes in crop pest management associated with the adoption of transgenic crops. As part of an IUPAC project on this topic, recent data are reviewed regarding the alterations in pesticide use that have been observed in practice. Most results indicate a decrease in the amounts of active ingredients applied to transgenic crops compared with conventional crops. In addition, a generic environmental indicator—the environmental impact quotient (EIQ)—has been applied by these authors and others to estimate the environmental consequences of the altered pesticide use on transgenic crops. The results show that the predicted environmental impact decreases in transgenic crops. With the advent of new types of agronomic trait and crops that have been genetically modified, it is useful to take also their potential environmental impacts into account. Copyright © 2007 Society of Chemical Industry
Organophosphate insecticides and acaricides antagonise bifenazate toxicity through esterase inhibition in Tetranychus urticae
Authors: Van Leeuwen Thomas , Van Pottelberge Steven , Nauen Ralf , Tirry Luc
Published: September 2007

BACKGROUND: Bifenazate is a carbazate acaricide known for its potency, particularly against tetranychid mite species such as the two‐spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae). It was recently shown that the compound needs to be activated by an S,S,S‐tributyl‐phosphorotrithioate (DEF)‐sensitive mechanism in spider mites to display full acaricidal efficacy. The ability of well‐known organophosphates and carbamates to inhibit the activation of bifenazate and thus compromise its acaricidal potential was tested.

RESULTS: Esterase activity determined in vivo after pre‐exposure of mites with organophosphates and carbamates revealed–depending on the compound–varying esterase inhibition nicely correlated with the ability of the individual compound to antagonise bifenazate action on mites.

CONCLUSIONS: The findings illustrate that organophosphates and carbamates interfere with bifenazate efficacy, most probably by inhibiting carboxylesterases responsible for the activation of the pro‐drug. As a result of the strong antagonism, mixtures of bifenazate with carbamates or organophosphates should not be used under field conditions. Moreover, there exists a real threat in repeatedly applying organophosphates and bifenazate. The present study again illustrates how important mode of action information is for the proper planning of resistance management strategies. Copyright © 2007 Society of Chemical Industry
Fmoc‐based solid phase chemical synthesis of 71‐meric neuregulin 1‐β1, an epidermal growth factor‐like domain
Authors: Kakizawa Taeko , Koide‐Yoshida Shizuyo , Kimura Tooru , Uchimura Hiromasa , Hayashi Yoshio , Saito Kazuki , Kiso Yoshiaki
Published: September 2007

The human neuregulin 1‐β1 (NRG1‐β1, amino acid residues 176–246) was chemically synthesized by Fmoc‐based solid phase peptide synthesis (SPPS) followed by folding in a redox buffer. The biological activity of the synthesized NRG1‐β1 was confirmed by ligand‐induced tyrosine phosphorylation on Chinese hamster ovary (CHO) cells expressing ErbB‐4. Copyright © 2007 European Peptide Society and John Wiley & Sons, Ltd.
A Bowman‐Birk trypsin inhibitor with antiproliferative activity from Hokkaido large black soybeans
Authors: Ho Vincent S. M. , NG T. B.
Published: September 2007

A trypsin inhibitor, with an N‐terminal sequence highly homologous to those of 8‐kDa Bowman‐Birk trypsin inhibitors, was isolated from the seeds of Hokkaido large black soybeans. The trypsin inhibitor was unadsorbed on SP‐Sepharose but adsorbed on DEAE‐cellulose and Mono Q. It inhibited proliferation in breast cancer (MCF‐7) cells and hepatoma (Hep G2) cells with an IC50 of 35 and 140 µM, respectively. The trypsin inhibitory activity of the inhibitor was completely preserved after exposure to temperatures up to 100 °C for 30 min and to the pH range 2–13 for the same duration. The trypsin inhibitor inhibited HIV‐1 reverse transcriptase with an IC50 of 38 µM, but was devoid of antifungal activity toward Fusarium oxysporum and Mycosphaerella arachidicola. Copyright © 2007 European Peptide Society and John Wiley & Sons, Ltd.
Effect of ZnS shell on the Raman spectra from CdSe nanorods
Authors: Lange Holger , Machón María , Artemyev Mikhail , Woggon Ulrike , Thomsen Christian
Published: September 2007

We investigated the influence of an epitaxially grown ZnS shell on the phonon spectra of CdSe nanorods of different sizes. The CdSe related Raman peaks shift with addition of a ZnS shell. The longitudinal optical phonon shifts slightly due to strain and the low‐energy shoulder shifts stronger, which can be explained within a model for surface optical phonons. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Ameliorative effects of pycnogenol® on carbon tetrachloride‐induced hepatic oxidative damage in rats
Authors: Ahn Tai‐Hwan , Yang Young‐Su , Lee Jong‐Chan , Moon Chang‐Jong , Kim Sung‐Ho , Jun Woojin , Park Seung‐Chun , Kim Jong‐Choon
Published: September 2007

This study evaluated the putative antioxidant activity of Pycnogenol® (PYC) against CCl4‐induced hepatic oxidative damage in Sprague‐Dawley rats. A single oral dose of CCl4 (1.25 mL/kg) produced significantly increased levels of serum aminotransferase (AST) and alanine aminotransferase (ALT) activities. In addition, increased malondialdehyde (MDA) concentration, reduced glutathione (GSH) content, and decreased catalase, superoxide dismutase (SOD) and glutathione‐S‐transferase (GST) activities were observed in the hepatic tissues. However, concomitant administration with PYC (10 or 20 mg/kg) significantly improved CCl4‐induced hepatic injury, as evidenced by the decline of serum AST and ALT activities in a dose dependent manner. Moreover, PYC reduced MDA concentration and increased GSH levels and catalase, SOD and GST activities in hepatic tissues, indicating that concomitant administration with PYC efficiently prevent the CCl4‐induced oxidative damage in rats. The free radical scavenging assay showed that PYC has a dose‐dependent scavenging activity against 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) free radicals. These results indicate that PYC has an antioxidant effect against CCl4‐induced hepatic oxidative damage and is useful as a hepatoprotective agent against various liver diseases induced by oxidative stress. Copyright © 2007 John Wiley & Sons, Ltd.
Effectiveness and safety of Devil's Claw tablets in patients with general rheumatic disorders
Authors: Warnock Mary , McBean Douglas , Suter Andreas , Tan Jen , Whittaker Patricia
Published: September 2007

Arthritis and other rheumatic conditions (AORC) are the leading cause of disability, are associated with poor quality of life and incur considerable direct and indirect costs. It is considered that the instance of AORC will continue to increase. To assess the effectiveness, safety and tolerability of Harpagophytum (Bioforce) in the treatment of AORC, a single group open study of 8 weeks duration (259 patients) was performed in the United Kingdom. Effectiveness was assessed by numeric rating scales, the Western Ontario and McMasters Universities Osteoarthritis (WOMAC) Index and the Algofunctional Hand Osteoarthritis Index. Tolerance was measured by a numeric rating scale and safety by self‐reporting, blood analysis and liver function tests. Quality of life was measured by SF‐12 questionnaire. There were statistically significant (p < 0.0001) improvements in patient assessment of global pain, stiffness and function. There were also statistically significant reductions in mean pain scores for hand, wrist, elbow, shoulder, hip, knee and back pain. Quality of life measurements (SF‐12) were significantly increased from baseline and 60% patients either reduced or stopped concomitant pain medication. Harpagophytum is an effective and well‐tolerated serious treatment option for mild to moderate degenerative rheumatic disorders providing improved quality of life measure. Copyright © 2007 John Wiley & Sons, Ltd.
Antiviral effects of Glycyrrhiza species
Authors: Fiore Cristina , Eisenhut Michael , Krausse Rea , Ragazzi Eugenio , Pellati Donatella , Armanini Decio , Bielenberg Jens
Published: September 2007

Historical sources for the use of Glycyrrhiza species include ancient manuscripts from China, India and Greece. They all mention its use for symptoms of viral respiratory tract infections and hepatitis. Randomized controlled trials confirmed that the Glycyrrhiza glabra derived compound glycyrrhizin and its derivatives reduced hepatocellular damage in chronic hepatitis B and C. In hepatitis C virus‐induced cirrhosis the risk of hepatocellular carcinoma was reduced. Animal studies demonstrated a reduction of mortality and viral activity in herpes simplex virus encephalitis and influenza A virus pneumonia. In vitro studies revealed antiviral activity against HIV‐1, SARS related coronavirus, respiratory syncytial virus, arboviruses, vaccinia virus and vesicular stomatitis virus.

Mechanisms for antiviral activity of Glycyrrhiza spp. include reduced transport to the membrane and sialylation of hepatitis B virus surface antigen, reduction of membrane fluidity leading to inhibition of fusion of the viral membrane of HIV‐1 with the cell, induction of interferon gamma in T‐cells, inhibition of phosphorylating enzymes in vesicular stomatitis virus infection and reduction of viral latency.

Future research needs to explore the potency of compounds derived from licorice in prevention and treatment of influenza A virus pneumonia and as an adjuvant treatment in patients infected with HIV resistant to antiretroviral drugs. Copyright © 2007 John Wiley & Sons, Ltd.