bluuebunny/crossref_metadata_2025 huggingface dataset:
Transferrin overexpression alters testicular function in aged mice
Authors: Lécureuil C. , Staub C. , Fouchécourt S. , Maurel M.C. , Fontaine I. , Martinat N. , Gauthier C. , Daudignon A. , Delaleu B. , Sow A. , Jégou B. , Guillou F.
Published: September 2006

Many studies have shown a correlation between transferrin (Tf) concentration and sperm yield in several mammalian species. We have used transgenic mice expressing human Tf (hTf) to investigate if overexpression of Tf increases the efficiency of mouse spermatogenesis. We demonstrated that a 36% increase of Tf does not ameliorate the efficiency of mouse spermatogenesis but on the contrary resulted in a 36% decrease of testis sperm reserves. Tf overexpression had no effect on testicular determination and development, however testicular function of these transgenic mice was affected in an age‐dependent manner. At 16 months of age, testicular and epididymal weights were significantly reduced. While spermatogenesis was qualitatively normal, testicular functions were perturbed. In fact, testosterone rate after human chorionic gonadotropin (hCG) stimulation was lower in Tf overexpressing mice. Intratesticular concentration of estradiol‐17β was increased and fluid accumulation after ligation of rete testis was more abundant in these transgenic mice. Surprisingly, we found that endogenous Tf levels were also increased in Tf overexpressing mice and we demonstrated for the first time that Tf may serve to upregulate its own expression in testis. Collectively, our data show that Tf overexpression has negative effects on testicular function and that Tf levels require strict regulation in the testis. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Computer‐aided meiotic maturation assay (CAMMA) of zebrafish (danio rerio) oocytes in vitro
Authors: Lessman Charles A. , Nathani Ravikanth , Uddin Rafique , Walker Jamie , Liu Jianxiong
Published: September 2006

We have developed a new technique called Computer‐Aided Meiotic Maturation Assay (CAMMA) for imaging large arrays of zebrafish oocytes and automatically collecting image files at regular intervals during meiotic maturation. This novel method uses a transparency scanner interfaced to a computer with macro programming that automatically scans and archives the image files. Images are stacked and analyzed with ImageJ to quantify changes in optical density characteristic of zebrafish oocyte maturation. Major advantages of CAMMA include (1) ability to image very large arrays of oocytes and follow individual cells over time, (2) simultaneously image many treatment groups, (3) digitized images may be stacked, animated, and analyzed in programs such as ImageJ, NIH‐Image, or ScionImage, and (4) CAMMA system is inexpensive, costing less than most microscopes used in traditional assays. We have used CAMMA to determine the dose response and time course of oocyte maturation induced by 17α‐hydroxyprogesterone (HP). Maximal decrease in optical density occurs around 5 hr after 0.1 ug/ml HP (28.5°C), approximately 3 hr after germinal vesicle migration (GVM) and dissolution (GVD). In addition to changes in optical density, GVD is accompanied by streaming of ooplasm to the animal pole to form a blastodisc. These dynamic changes are readily visualized by animating image stacks from CAMMA; thus, CAMMA provides a valuable source of time‐lapse movies for those studying zebrafish oocyte maturation. The oocyte clearing documented by CAMMA is correlated to changes in size distribution of major yolk proteins upon SDS–PAGE, and, this in turn, is related to increased cyclin B1 protein. Mol. Reprod. Dev. 74: 97–107, 2007. © 2006 Wiley‐Liss, Inc.
Embryo quality and production efficiency of porcine parthenotes is improved by phytohemagglutinin
Authors: Gupta Mukesh Kumar , Uhm Sang Jun , Han Dong Wook , Lee Hoon Taek
Published: September 2006

In vitro production of porcine embryos has become routine in most laboratories but the yield and quality of the resultant blastocysts remain suboptimal. Phytohemagglutinin (PHA) is an N‐acetylgalactosamine/galactose sugar‐specific lectin with a wide variety of biological activities including mitogenesis, mediation of cell recognition, and agglutination of cells. This study was therefore, designed to investigate the effect of PHA on the preimplantation embryo development and quality of in vitro produced porcine parthenotes. Parthenogenetic presumptive diploid zygotes were produced in vitro by electrical activation and cultured in the absence or presence of PHA at different concentrations (0, 5, 10, 15, 20 µg/ml). There were no significant differences in the cleavage rate of porcine parthenotes in control and treatment groups at all tested concentrations of PHA (P < 0.05). However, supplementation of PHA at the concentration of 15 µg/ml significantly improved the blastocyst rate (68.9 ± 1.5% vs. 43.1 ± 4.1%), hatching rate (25.8 ± 3.1% vs. 8.9 ± 2.0%), and total nuclei number (95.5 ± 9.3 vs. 63.4 ± 4.3) when compared to control group (P > 0.05). TUNEL labeling revealed that blastocysts in PHA group were less predisposed to biochemical apoptosis than in control group while total apoptosis and nuclear fragmentation remained unaltered. Real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) analysis further revealed that PHA decreased the expression ratio of BAX/BCL‐XL and enhanced the relative abundance of IGF2 transcripts. Therefore, our study suggests that PHA improves the blastocyst yield and quality by enhancing blastocyst expansion, hatching, and total cell number and decreasing the apoptosis by positively modulating the expression of embryo survival related genes. Mol. Reprod. Dev. 74: 435–444, 2007. © 2006 Wiley‐Liss, Inc.
Changes of maternal transcripts in oocytes from persistent follicles in cattle
Authors: Lingenfelter Brandon M. , Dailey Robert A. , Inskeep E. Keith , Vernon Michael W. , Poole Daniel H. , Rhinehart Justin D. , Yao Jianbo
Published: September 2006

A high incidence of early embryonic loss is associated with prolonged dominance of follicles. The objective of the present experiment was to determine if persistence of a follicle resulted in alterations in mRNA expression of important genes in the oocyte. Cows were assigned to four groups: growing follicles on day 6 (G0h) or day 8 (G48h) and persistent follicles on day 13 (P0h) or day 15 (P48h) of the estrous cycle (estrus = day 0). All cows were super‐stimulated on day 1–4. Cows in G48h, P0h, and P48h groups received 25 mg prostaglandin (PG) F2α on day 6. Cows in P0h and P48h groups received progesterone from CIDR‐B devices on day 5 through 13. Ovaries of cows in G0h, G48h, P0h, and P48h groups were removed on day 6, 8, 13, and 15, respectively. Oocytes were aspirated immediately after colpotomy and denuded of cumulus cells. Quantitative real‐time PCR was used to measure the mRNA abundances of 10 selected genes important for early embryogenesis in oocytes obtained from growing and persistent follicles. Relative abundances of MSY2, PARN, and YY1 mRNA (P < 0.05) were significantly lower in oocytes from persistent than from growing follicles. Oocytes from persistent follicles, however, had greater abundances of PAP and eIF‐4E transcripts (P < 0.05). The data indicate that persistence of a follicle leads to altered abundances of mRNA for genes important for regulation of transcription and protein translation in the oocyte, which could compromise development of early embryos in cows that ovulate a persistent follicle. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Impact of in vitro production techniques on the expression of X‐linked genes in bovine (bos taurus) oocytes and pre‐attachment embryos
Authors: Nino‐Soto Maria I. , Basrur Parvathi K. , King W. Allan
Published: September 2006

Our previous studies showed that expression patterns of X‐linked genes in cultured cells are different from those of their tissues of origin. This investigation analyses the transcription pattern of the X‐linked genes BIRC4, GAB3, MECP2, RPS4X, SLC25A6, and XIST in bovine in vitro matured oocytes and in vitro fertilized embryos, and their in vivo counterparts. In vitro‐derived pools of mature oocytes and pre‐attachment embryos were obtained by: (a) TCM‐199/serum with bovine oviductal epithelial cells as co‐culture, and (b) synthetic oviductal fluid/BSA. Pools of in vivo‐derived morulae and blastocysts were provided by a commercial embryo transfer operation. Total RNA was extracted for quantification of gene‐specific transcript levels using real‐time quantitative PCR. Statistical analysis was performed using a mixed model factorial ANOVA with α = 0.05. The effect of the in vitro environmental conditions on X‐linked gene transcription was most evident during the fourth cell cycle, at the period of activation of the embryonic genome, and seemed to be less pronounced at later developmental stages, with the exception of BIRC4. The levels of X‐linked genes transcripts in in vivo‐derived embryos were lower relative to their in vitro counterparts for all genes tested. Finally, the pattern of expression of XIST in bovine oocytes and embryos was similar to that reported in humans. These results highlight the possibility that X‐linked gene expression analysis is a useful tool to monitor the impact of reproductive biotechnologies on the developmental potential of embryos and aid in their improvement. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
F‐actin involvement in guinea pig sperm motility
Authors: Azamar Yenia , Uribe Salvador , Mújica Adela
Published: September 2006

Sperm motility is a must for natural fertilization to occur. During their travel through the epididymis, mammalian spermatozoa gradually acquire the ability to move. This is accomplished through a sliding movement of the outer doublet microtubules of the axoneme which is energized by the dynein ATPase. Within its complex structure, the mammalian sperm flagellum contains F‐actin and thus, we decided to test in the guinea pig sperm flagellum the role of F‐actin in motility. During maturation, capacitation, and the acrosome reaction, a gradual decrease of the relative concentration of F‐actin was observed. Motility increased as spermatozoa became able to fertilize. Gelsolin, phalloidin, and KI inhibited sperm motility. Gelsolin canceled sperm motility within 20 min of treatment while 0.6 M KI had immediate effects. Phalloidin diminished hyperactive sperm motility slightly. All three compounds significantly increased the relative concentration of F‐actin. Latrunculins are conventional drugs that destabilize the F‐actin cytoskeleton. Latrunculin A (LAT A) did not affect sperm motility; but significantly increased F‐actin relative concentration. The results suggested that in guinea pig spermatozoa, randomly severing F‐actin filaments inhibits flagellar motility; while end filament alteration does not. Thus, specific filament regions seem to be important for sperm motility. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Targeting the exogenous htPAm gene on goat somatic cell beta‐casein locus for transgenic goat production
Authors: Shen Wei , Lan Guocheng , Yang Xueyi , Li Lan , Min Lingjiang , Yang Zhengtian , Tian Liyuan , Wu Xiaojie , Sun Yujiang , Chen Hong , Tan Jinghe , Deng Jixian , Pan Qingjie
Published: September 2006

Combining gene targeting of animal somatic cells with nuclear transfer technique has provided a powerful method to produce transgenic animal mammary gland bioreactor. The objective of this study is to make an efficient and reproducible gene targeting in goat fetal fibroblasts by inserting the exogenous htPAm cDNA into the beta‐casein locus with liposomes or electroporation so that htPAm protein might be produced in gene‐targeted goat mammary gland. By gene‐targeting technique, the exogenous htPAm gene was inserted to milk goat beta‐casein gene sequences. Fetal fibroblasts were isolated from Day 35 fetuses of Guanzhong milk goats, and transfected with linear gene‐targeting vector pGBC4htPAm using Lipefectamin™‐2000 and electoporation, respectively. Forty‐eight gene‐targeted cell colonies with homologous recombination were obtained, and three cell colonies were verified by DNA sequence analysis within the homologous recombination region. Using gene‐targeted cell lines as donor cells for nuclear transfer, a total of 600 reconstructed embryos had been obtained, and 146 developed cloned embryos were transferred to 16 recipient goats, and finally three goats showed pregnancy at Day 90. Mol. Reprod. Dev. 74: 428–434, 2007. © 2006 Wiley‐Liss, Inc.
Plasticity of epidermal adult stem cells derived from adult goat ear skin
Authors: Yang Xueyi , Qu Lei , Wang Xin , Zhao Ming , Li Wei , Hua Jinlian , Shi Mingyan , Moldovan Nicanor , Wang Hongfeng , Dou Zhongying
Published: September 2006

Here we report the isolation and characterization of pluripotent stem cells from adult goat skin. We found that these primary cells have the properties of embryonic stem cells (ESC), including the expression of appropriate immunological markers and the capability of forming embryoid bodies. The subcultured cells also show the characteristics of stem cells, such as the expression of CK19, β1‐integrin, P63, and formation of holo‐clones in culture. Therefore, we termed these cells epidermal adult stem cells (EpiASC), although their origin was not identified. We have shown that clones of individual EpiASC proliferate and differentiate in culture to produce neurons, cardiomyocytes, osteoblasts, and occytes. Further, we cultivated EpiASC on bioengineered dermis and denuded human amniotic membrane (HAM), to reconstruct artificial skin and corneal epithelium. We successfully transplanted those artificial tissues in goats with acute full‐thickness skin defect (AFTSD) and limbal stem cell deficiency (LSCD), respectively. Our results showed that indeed EpiASC reconstructed the skin (hair was observed in restored areas), and repaired the damaged cornea of goats with total LSCD. These data confirm that EpiASC can differentiate into different functional cell types in vivo or in vitro. Due to their high degree of inherent plasticity, and to their easy accessibility for collection from the skin, EpiASC are excellent candidate sources for diverse cell therapies. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Sex‐specific windows for high mRNA expression of DNA methyltransferases 1 and 3A and methyl‐CpG‐binding domain proteins 2 and 4 in human fetal gonads
Authors: Galetzka Danuta , Weis Eva , Tralau Tim , Seidmann Larissa , Haaf Thomas
Published: September 2006

DNA methyltransferases (DNMTs) and 5‐methyl‐CpG‐binding domain proteins (MBDs) are involved in the acquisition of parent‐specific epigenetic modifications in human male and female germ cells. Reverse Northern blot analyses demonstrated sex‐specific differences in mRNA expression for the maintenance DNMT1 and the de novo DNMT3A in developing testis and ovary. In fetal testis DNMT1 and DNMT3A expression peaked in mitotically arrested spermatogonia around 21 weeks gestation. In fetal ovary transcriptional upregulation of DNMT1 and DNMT3A occurred during a very brief period at 16 weeks gestation, when the oocytes proceeded through meiotic prophase. Fetal gonads showed several fold higher DNMT3A expression levels than fetal brain and adult tissues. The most abundant DNMT3A isoform in fetal testis and ovary was DNMT3A2, whereas in all other analyzed tissues DNMT3A1 predominated. The catalytically inactive DNMT3A3 isoform was also present at relatively high levels in developing gonads and may perform a regulatory function(s). In both male and female fetal gonads expression of genes for MBD2 and MBD4, which may be implicated in chromatin remodeling of methylated genomic DNA sequences, was tightly linked to DNMT expression. We propose that the sex‐specific time windows for concomitant upregulation of DNMT1, DNMT3A, MBD2, and MBD4 are associated with prenatal remethylation of the human male and female germ line. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Differential expression of oxygen‐regulated genes in bovine blastocysts
Authors: Harvey A.J. , Santos A. Navarrete , Kirstein M. , Kind K.L. , Fischer B. , Thompson J.G.
Published: September 2006

Low oxygen conditions (2%) during post‐compaction culture of bovine blastocysts improve embryo quality, which is associated with a small yet significant increase in the expression of glucose transporter 1 (GLUT‐1), suggesting a role of oxygen in embryo development mediated through oxygen‐sensitive gene expression. However, bovine embryos to at least the blastocyst stage lack a key regulator of oxygen‐sensitive gene expression, hypoxia‐inducible factor 1α (HIF1α). A second, less well‐characterized protein (HIF2α) is, however, detectable from the 8‐cell stage of development. Here we use differential display to determine additional gene targets in bovine embryos in response to low oxygen conditions. While development to the blastocyst stage was unaffected by the oxygen concentration used during post‐compaction culture, differential display identified oxygen‐regulation of myotrophin and anaphase promoting complex 1 expression, with significantly lower levels observed following culture under 20% oxygen than 2% oxygen. These results further support the hypothesis that the level of gene expression of specific transcripts by bovine embryos alters in response to changes in the oxygen environment post‐compaction. Specifically, we have identified two oxygen‐sensitive genes that are potentially regulated by HIF2 in the bovine blastocyst. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Molecular identities of human sperm proteins reactive with antibodies in sera of immunoinfertile women
Authors: Bhande Satish , Naz Rajesh K.
Published: September 2006

Antisperm antibodies (ASA) can cause infertility in both men and women. It is important to delineate the sperm antigens against which these ASA are directed. Sperm proteins were separated by 2D gel electrophoresis and transferred to nitrocellulose membrane and incubated with sera from fertile women or immunoinfertile women having ASA. The corresponding immunoreactive peptide spots were cored from the gel and analyzed by the two‐dimensional (2D) gel electrophoresis/matrix‐assisted laser desoprtion ionization‐time of flight‐mass spectrometry and liquid chromatography‐mass spectrometry (MALDI‐TOF‐MS/LC‐MS). A total of 68 spots belonging to 38 different proteins and their isomers were identified. Fourteen of these proteins and their isomers reacted with both the fertile and immunoinfertile sera. Twenty‐four of these proteins reacted specifically only with the immunoinfertile sera and not with the fertile sera. Among them was a novel protein designated as a hypothetical protein FLJ32704 (accession # Q96MA6). An immunodominant sequence (amino acid 151‐159) of this protein was identified and a nonamer peptide based upon this sequence (IQTLG1TPR) was synthesized and examined for its immunoreactivity. This synthetic peptide reacted with 90% (36/40) of immunoinfertile sera and not with any of the fertile sera (0/40) in the enzyme‐linked immnosorbent assay (ELISA). In conclusion, using the 2D gel electrophoresis/MALDI‐TOF‐MS/LC‐MS procedure, we have identified several known and at least one novel antigen against which the antibodies are present in sera of immunoinfertile but not fertile women. Some of these antigens may find applications in specific diagonsis and treatment of infertility/immunoinfertility, and in the development of new generation of contraceptive modalities including contraceptive vaccines. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Oxidative stress reduces the muscarinic receptor function in the urinary bladder
Authors: de Jongh Rik , Haenen Guido R.M.M. , van Koeveringe Gommert A. , Dambros Miriam , De Mey Jo G.R. , van Kerrebroeck Philip E.V.
Published: September 2006

Aims

Several pathophysiological conditions in the urinary bladder, for example, ischemia/reperfusion and inflammation are characterized by the formation of reactive oxygen species (ROS). The ROS are highly toxic because they can destroy proteins, DNA, and lipids. The aim of this study was to investigate the effect of oxidative stress on excitation‐contraction coupling of detrusor smooth muscle.

Materials and Methods

Smooth muscle strips were dissected from pig urinary bladder and mounted in organ baths. Oxidative stress was mimicked by the addition of Cumene hydroperoxide (CHP), a lipophilic hydroperoxide, to the organ baths. Contractile responses to electrical field stimulation (EFS: 4–32 Hz), carbachol (10−8–3 × 10−5 M), potassium (65.3 mM), and ATP (1 mM) were monitored before and after the addition of CHP.

Results

Responses of detrusor strips to EFS were for the greater part based on neurogenic stimulation and the release of acetylcholine. CHP diminished contractile responses to EFS and carbachol to the same extent. The pD2 value of the carbachol concentration‐response curve decreased significantly after exposure to 0.1 mM, 0.4 mM, 0.8 mM CHP. Furthermore the maximal effect obtained with carbachol was significantly reduced after 0.1 mM, 0.4 mM, and 0.8 mM CHP treatment. Contractions induced by potassium and ATP were significantly less affected by oxidative stress compared to EFS‐ and carbachol‐induced responses of comparable amplitude.

Conclusions

The results of our study demonstrate that oxidative stress induced by CHP affects pig bladder contractility. The muscarinic receptor signaling system is severely damaged. L‐type calcium channels and the contractile system are less affected and cholinergic nerves remain largely unaffected. Neurourol. Urodynam. 26:302–308, 2007. © 2006 Wiley‐Liss, Inc.
Preoperative erectile function is one predictor for post prostatectomy incontinence
Authors: Wille S. , Heidenreich A. , Hofmann R. , Engelmann U.
Published: September 2006

Aims

The precise etiology of post prostatectomy incontinence (PPI) is not fully understood and risk factors are not yet comprehensively defined. It has been reported that sparing of the neurovascular bundle during prostatectomy improves postoperative erectile function, whereas the influence on urinary control is unclear. From daily clinical experience we made the impression that patients who are in the best shape have better erections and better continence. We therefore searched our database for a possible correlation between the preoperative erectile function and the incidence of PPI.

Patients and Methods

Four hundred three patients who underwent radical retropubic prostatectomy between January 2000 and May 2003 were enrolled into this retrospective study. Data of 327 patients (response rate 81%) at a median follow‐up of 26 months were analyzed using the validated International Index of Erectile Function (IIEF 5), the validated Urinary Distress Inventory (UDI6) and a standardized urinary symptom inventory. Continence was defined as usage of no or one pad daily. Erectile Dysfunction (ED) was defined as none/mild or moderate/severe with an IIEF 5 score of 17 or more or less than 17, respectively.

Results

Univariate and mulitvariate logistic regression analysis including preoperative IIEF 5 scores, age and nerve sparing prostatectomy, identified preoperative erectile function as significant predictor for PPI (P = 0.024), whereas age (P = 0.759) and nerve sparing prostatectomy (P = 0.504) did not predict PPI.

Conclusion

Erectile function is a predictor of PPI and should be recorded preoperatively. Neurourol. Urodynam. © 2006 Wiley‐Liss, Inc.
Long‐term patient satisfaction following cadaveric pubovaginal sling incontinence surgery using the UDI and IIQ‐7 questionnaires
Authors: Pianezza Michael L. , Joffe Robert , Chugh Taher , Radomski Sidney B.
Published: September 2006

Aims

The goal of any incontinence surgery is overall long‐term patient satisfaction. The purpose of our study was to assess long‐term patient satisfaction following cadaveric fascia lata pubovaginal sling surgery using the urogenital distress inventory (UDI) and the short form of the incontinence impact questionnaire (IIQ‐7).

Materials and Methods

A total of 47 patients who underwent pubovaginal sling incontinence surgery with a minimum follow‐up of 2 years were sent by mail the UDI (total score = 300) and the IIQ‐7 (total score = 100). Thirty‐seven responses (78.7%) were obtained, with a mean follow‐up of 3.8 years (range 2.0–6.2).

Results

Of the 37 patients who responded, the mean UDI score was 75.8 and the mean IIQ‐7 score was 21.4. There was no difference in the mean IIQ‐7 score for patients with more than 4 years follow‐up as compared to the entire group (28.8, P = 0.22). In contrast, the mean UDI score for patients with more than 4 years follow‐up was greater compared to the entire group (99.1, P = 0.04). UDI subscale analysis revealed that patients mainly complained of both irritative and stress symptoms as opposed to obstructive/discomfort symptoms (P < 0.01). Patients with pre‐operative mixed incontinence had greater mean UDI and IIQ‐7 scores compared to patients with pre‐operative pure stress incontinence (96.7 vs. 58.0, P = 0.04; 32.5 vs. 11.9, P = 0.03).

Conclusions

Overall long‐term scores assessing quality of life (IIQ‐7) were good and those assessing symptom distress (UDI) were satisfactory following pubovaginal sling surgery. Patients with pre‐operative mixed incontinence are at greatest risk for post‐operative dissatisfaction. Neurourol. Urodynam. 26:185–189, 2007. © 2006 Wiley‐Liss, Inc.
Age‐related variation in isotopic indicators of diet at medieval Kulubnarti, Sudanese Nubia
Authors: Turner B. L. , Edwards J. L. , Quinn E. A. , Kingston J. D. , Van Gerven D. P.
Published: September 2006

This study compares trends in dietary composition in two large cemetery populations from the site of Kulubnarti (AD 550–800) in Sudanese Nubia. Bone collagen and bone apatite carbonate were analysed to characterise stable carbon, nitrogen and oxygen isotopes. Previous research on these cemeteries has suggested marked differences in nutritional status and health between the populations. Contrary to expectations, there were no significant relationships between any isotopic indicators related to sex or cemetery of burial, suggesting no isotopically‐measurable differences in diet. However, collagen δ13C and δ15N were significantly related to age, suggesting age‐related differences in protein intake or other factors.

Weaning trends are gradual and variable, with the range in δ15N values exceeding 4‰ among infants/young children (0–3 yrs) and standard deviations exceeding 1‰ in collagen δ13C and δ15N for both infants/young children and subadults (4–17 yrs). This suggests varied weaning strategies among both populations and variable diets prior to adulthood. Also observed was a distinct range of isotopic carbon and nitrogen values among individuals classified as subadults (4–17 yrs), who are depleted in collagen δ13C and δ15N relative to adults. However, both infants/young children and subadults are slightly enriched in δ18O relative to adults, which suggests the presence of non‐local individuals or age‐related variation in water sources. While most isotopic studies of age‐related dietary trends have focused on reconstructing the weaning process, this study presents findings that indicate tripartite isotopic trends distinguishing infancy, subadulthood and adulthood as separate dietary categories. Broad similarities are evident between the results presented here and those from several earlier studies of smaller populations and to nutritional studies of modern communities. These findings suggest that further research into health disparities at Kulubnarti should focus on non‐dietary causal factors, and more generally, that greater attention should be paid to subadulthood in palaeodiet studies. Copyright © 2006 John Wiley & Sons, Ltd.
Diagnostic osteology and analysis of the Mid‐ to Late Holocene dynamics of shags and cormorants in Tierra del Fuego
Authors: Causey D. , Lefèvre C.
Published: September 2006

We present here illustrated characteristics and anatomical descriptions of features that can be used to discriminate between four common skeletal elements (i.e. humerus, coracoid, femur, tarsometatarsus) of the five species of shags and cormorants known to occur in southern South America. We also present a detailed study of their distribution and abundance from about 6000 years before present to historical times as revealed by identification of faunal material excavated earlier and by re‐analysis of material published previously. Our results present a high‐resolution examination of the avian resource base used by early human hunters, and provide a foundation for future studies on the palaeoavifauna of Tierra del Fuego during the Mid‐ to Late Holocene. On the broadest scales, species diversity of the Phalacrocoracidae is qualitatively stable over space and time, a pattern that is also reflected in the larger marine bird community. On a finer scale, however, our results indicate that the abundance and distribution of cormorants and shags in Mid‐ and Late Holocene zooarchaeological deposits varied in a complex manner through time. These patterns do not appear to be related to proximity effects of hunters to colonies, but to other factors possibly associated with environmental change. Copyright © 2006 John Wiley & Sons, Ltd.
Estimation of adult skeletal age‐at‐death: statistical assumptions and applications
Authors: Samworth R. , Gowland R.
Published: September 2006

We examine the statistical assumptions underlying different techniques of estimating the age‐at‐death of a skeleton from one or more age indicators. The preferred method depends on which property of the distribution of the data in the reference sample is preserved in the skeleton to be aged. In cases where the conditional distribution of age given indicator is preserved, we provide ‘look‐up’ tables giving essentially unbiased age estimates and prediction intervals, using a large reference sample and the auricular surface and pubic symphysis age indicators. Where this assumption is violated, but the conditional distribution of indicator given age is preserved, we find that an alternative model which attempts to capture the biological process of development of an individual has some attractive features, which may make it suitable for further study. Copyright © 2006 John Wiley & Sons, Ltd.
An optimal regulator for linear systems with multiple state and input delays
Authors: Basin Michael , Perez Joel
Published: September 2006

This paper presents an optimal regulator for a linear system with multiple state and input delays and a quadratic criterion. The optimal regulator equations are obtained reducing the original problem to the linear‐quadratic regulator design for a system without delays. Performance of the obtained optimal regulator is verified in the illustrative example against the best linear regulators available for the linear system without delays and for two rational approximations of the original time‐delay system. Simulation graphs demonstrating better performance of the obtained optimal regulator with respect to the criterion value are included. Copyright © 2006 John Wiley & Sons, Ltd.
Comparative study of new thermotropic polyazomethines
Authors: Marin Luminita , Cozan Vasile , Bruma Maria
Published: September 2006

Three series of novel modified polyazomethines were prepared with mesogenic units and varying kinking groups. The effect of the incorporation of the kinking groups on the thermotropic properties of the alternated and random polymers was studied. Structural elucidation was carried out by elemental analysis, infrared (IR), proton nuclear magnetic resonance (1H‐NMR) and UV‐vis spectroscopy. Inherent viscosity of the polymers measured in N,N‐dimethyl formamide (DMF) at 20°C was in the range 0.15–0.44 dl/g. Their molecular weight and molecular weight distribution were determined using gel permeation chromatography (GPC). Mesomorphic properties were studied by differential scanning calorimetry (DSC), hot stage polarizing optical microscopy and the thermal stability was determined by thermogravimetric analysis (TGA). Some polyazomethines exhibited fine grain texture and good mesophase stability. It was observed that the random copolymers had lower melting point and higher mesophase stability than alternated polymers. Copyright © 2006 John Wiley & Sons, Ltd.
Biopolymer‐hydroxyapatite composite coatings prepared by electrospinning
Authors: Bishop Aisha , Balázsi Csaba , Yang Jason H. C. , Gouma Pelagia‐Irene
Published: September 2006

Biocompatibility in research and development of advanced prosthetics is a current problem faced by medical researchers. Hydroxyapatite (HA) is a natural ceramic (responsible for strength and stability in the human skeletal system) operable as a biocomposite coating to improve the biocompatibility of materials such as silicon nitride, typically used for bone replacement. In this study, polymer based composites were prepared by electrospinning HA with a biocompatible polymer for the development of a structurally stable casing for prosthetic devices. Presented in this paper is the morphological and structural study of electrospun polymer‐HA mats. Electrospun mats were calcinated at the phase transformation temperature of HA revealing pure HA wires with diameters just under 1 µm and lengths of the order of 10 to 50 µm. Results have outlined the dispersion mechanism of HA electrospun with polyvinylpyrrolidone (PVP) and cellulose acetate (CA) in varying solvents to discern the effects of acidic additives and electrospinning on the production of HA wires. Copyright © 2006 John Wiley & Sons, Ltd.
High resolution proteome/peptidome analysis of body fluids by capillary electrophoresis coupled with MS
Authors: Schiffer Eric , Mischak Harald , Novak Jan
Published: September 2006

All organisms contain thousands of proteins and peptides in their body fluids. A deeper insight into the functional relevance of these polypeptides under different physiological and pathophysiological conditions and the discovery of specific peptide biomarkers would greatly enhance both diagnosis and therapy of specific diseases. Proteomic methods can provide means to accomplish this grand medical vision. In this review, we will focus on the potential use of proteome analysis for clinical applications, such as disease diagnosis and assessment of response to therapy. We focus on CE coupled with MS (CE‐MS) and review in detail different aspects of CE‐MS coupling and the results obtained using CE‐MS analysis of clinically relevant samples. We also discuss clinical applications of the technology for the diagnosis of renal diseases, urogenital cancer, and arteriosclerosis as well as monitoring the responses to therapeutic interventions.
Rejuvenating rice proteomics: Facts, challenges, and visions
Authors: Agrawal Ganesh Kumar , Jwa Nam‐Soo , Iwahashi Yumiko , Yonekura Masami , Iwahashi Hitoshi , Rakwal Randeep
Published: September 2006

Proteomics is progressing at an unprecedented pace, as can be exemplified by the progress in model organisms such as yeast, bacteria, and mammals. Proteomics research in plants, however, has not progressed at the same pace. Unscrambling of the genome sequences of the dicotyledoneous Arabidopsis thaliana (L.) and monocotyledoneous rice (Oryza sativa L.) plant species, respectively, has made them accessible reference organisms to study plant proteomics. Study of these two reference plants is expected to unravel the mystery of plant biology. Rice, a critically important food crop on the earth, has been termed a “cornerstone” and the “Rosetta stone” for functional genomics of cereal crops. Here, we look at the progress in unraveling rice proteomes and present the facts, challenges, and vision. The text is divided into two major parts: the first part presents the facts and the second part discusses the challenges and vision. The facts include the technology and its use in developing proteomes, which have been critically and constructively reviewed. The challenges and vision deal with the establishment of technologies to exhaustively investigate the protein components of a proteome, to generate high‐resolution gel‐based reference maps, and to give rice proteomics a functional dimension by studying PTMs and isolation of multiprotein complexes. Finally, we direct a vision on rice proteomics. This is our third review in series on rice proteomics, which aims to stimulate an objective discussion among rice researchers and to understand the necessity and impact of unraveling rice proteomes to their full potential.
Ions of the interactome: The role of MS in the study of protein interactions in proteomics and structural biology
Authors: Downard Kevin M.
Published: September 2006

The role of MS in the study of protein‐protein interactions in solution is described from a proteomics perspective, in terms of high‐throughput analyses of protein complexes in vivo, through to chemical and biochemical treatments ahead of MS analysis in the context of complementary experimental approaches in structural biology. The use of MS to characterise protein‐protein interactions is described following the single and tandem affinity purification of protein complexes and assemblies of expressed proteins in host cells, the isolation and preservation of protein complexes on surfaces and microarrays, and their prior treatment with chemical and biochemical probes by hydrogen exchange, radical probe, chemical cross‐linking, and limited proteolysis. The advantages and disadvantages of each of the approaches are presented. These new and emerging applications, which further demonstrate the power of MS, continue to ensure that the mass spectrometer will remain at the heart of discoveries in proteomics in the foreseeable future.
Plant proteome analysis: A 2004–2006 update
Authors: Rossignol Michel , Peltier Jean‐Benoît , Mock Hans‐Peter , Matros Andrea , Maldonado Ana M. , Jorrín Jesús V.
Published: September 2006

Since the appearance of the review entitled “Plant Proteome Analysis” in Proteomics in February 2004 (Cánovas, F. M., Dumas‐Gaudot, E., Recorbert, G., Jorrín, J. et al., Proteomics 2004, 4, 285–298), about 200 original articles focusing on plant proteomics have been published. Although this represents less than 1% of the global proteomics output during this period, it nevertheless reflects an increase in activity over the period 1999–2004. These papers concern the proteome of at least 35 plant species but have concentrated mainly on thale cress (Arabidopsis thaliana) and rice (Oryza sativa). The scientific objectives have ranged from a proteomic analysis of organs, tissues, cell suspensions, or subcellular fractions to the study of plant development and response to various stresses. A number of contributions have covered PTMs and protein interactions. The dominant analytical platform has been 2‐DE coupled to MS, but “second generation” techniques such as DIGE, multidimensional protein identification technology, isotope‐coded affinity tags, and stable isotope labeling by amino acids in cell culture have begun to make an impact. This review aims to provide an update of the contribution of proteomics to plant biology during the period 2004–2006, and is divided into six sections: introduction, subcellular proteomes, plant development, responses to biotic and abiotic stresses, PTMs, and protein interactions. The conclusions summarize a view of the major pitfalls and challenges of plant proteomics.
Proteome informatics I: Bioinformatics tools for processing experimental data
Authors: Palagi Patricia M. , Hernandez Patricia , Walther Daniel , Appel Ron D.
Published: September 2006

Bioinformatics tools for proteomics, also called proteome informatics tools, span today a large panel of very diverse applications ranging from simple tools to compare protein amino acid compositions to sophisticated software for large‐scale protein structure determination. This review considers the available and ready to use tools that can help end‐users to interpret, validate and generate biological information from their experimental data. It concentrates on bioinformatics tools for 2‐DE analysis, for LC followed by MS analysis, for protein identification by PMF, by peptide fragment fingerprinting and by de novo sequencing and for data quantitation with MS data. It also discloses initiatives that propose to automate the processes of MS analysis and enhance the quality of the obtained results.
Proteome informatics II: Bioinformatics for comparative proteomics
Authors: Lisacek Frédérique , Cohen‐Boulakia Sarah , Appel Ron D.
Published: September 2006

The present review attempts to cover the most recent initiatives directed towards representing, storing, displaying and processing protein‐related data suited to undertake “comparative proteomics” studies. Data interpretation is brought into focus. Efforts invested into analysing and interpreting experimental data increasingly express the need for adding meaning. This trend is perceptible in work dedicated to determining ontologies, modelling interaction networks, etc. In parallel, technical advances in computer science are spurred by the development of the Web and the growing need to channel and understand massive volumes of data. Biology benefits from these advances as an application of choice for many generic solutions. Some examples of bioinformatics solutions are discussed and directions for on‐going and future work conclude the review.
Safety of endobronchial biopsy in children with cystic fibrosis
Authors: Molina‐Teran A. , Hilliard T.N. , Saglani S. , Haxby E. , Scallan M. , Bush A. , Davies J.C.
Published: September 2006

There is little found in the published literature regarding the use of endobronchial biopsy (EBB) in children with cystic fibrosis (CF). One concern over the use of the technique may relate to safety, in particular increased risk of bleeding from a hypertrophied bronchial circulation. The aim of this retrospective study was to compare the safety of EBB in children with CF and those with other conditions, the most frequent of which included primary ciliary dyskinesia and recurrent lower respiratory tract infections. Case notes of all children undergoing EBB in our institution between February 2003 and May 2004 were reviewed. EBB was performed during 45 bronchoscopies in 42 CF patients (19 males, group mean age 7.13 ± 4.48 years) and in 39 controls (20 males, group mean age 6.59 ± 4.48 years). There were no significant differences between disease groups in the number, type, or severity of complications occurring during or in the first 12 hr after the procedure. We conclude that EBB performed as part of fibreoptic bronchoscopy (FOB) under general anaesthesia can be performed safely in children with CF, when both bronchoscopist and anaesthetist are suitably experienced. Studies of such samples would allow us to determine the early pathological changes in the CF airway and possibly find new treatments to prevent the progression to bronchiectasis and end stage airway destruction. Pediatr Pulmonol. 2006, 41:1021–1024. © 2006 Wiley‐Liss, Inc.
Passive respiratory mechanics measured during natural sleep in healthy term neonates and infants up to 8 weeks of life
Authors: Katier N. , Uiterwaal C.S.P.M. , de Jong B.M. , Verheij T.J.M. , van der Ent C.K.
Published: September 2006

The single occlusion technique (SOT) is a simple and noninvasive technique for measurement of passive respiratory mechanics in infants. Reference values based on measurements of a large population of healthy infants performed outside specialized research laboratories are lacking. The aim of this study was to present reference values for passive respiratory mechanics based on a large population of healthy term neonates and infants measured during natural sleep in routine care. As part of the ongoing Wheezing Illnesses Study Leidsche Rijn (WHISTLER), the compliance (Crs) and resistance (Rrs) of the respiratory system were measured in 450 healthy unsedated neonates and infants with a mean age of 4.6 ± 1.3 weeks. Multivariable regression analysis, with gestational age, age at measurement, body size, sex, and ethnicity as possible predictors, was carried out to estimate prediction equations for mean Crs and Rrs values. Technically acceptable lung function measurements could be performed in 328 (73%) neonates and infants. Median Crs was 39.5 (range 14.8–79.1) ml/kPa and median Rrs was 7.4 (range 3.8–19.5) kPa/L/sec. The following regression equations for Crs and Rrs were obtained: ln Crs = 1.677 + 1.3 × 10−4 × birth weight (g) + 0.030 × birth length (cm) and ln Rrs = 2.496–3.1 × 10−6 × birth length3 (cm3) − 0.114 × sex. We provided reference values for passive respiratory mechanics using the SOT in a large population of healthy term neonates and infants measured during natural sleep. These data provide a frame of reference for assessing the normality of SOT measurements performed in routine care. Pediatr Pulmonol. 2006, 41:1058–1064. © 2006 Wiley‐Liss, Inc.
The homozygous P582S mutation in the oxygen‐dependent degradation domain of HIF‐1α is associated with increased risk for prostate cancer
Authors: Orr‐Urtreger Avi , Bar‐Shira Anat , Matzkin Haim , Mabjeesh Nicola J.
Published: September 2006

BACKGROUND

The heterodimeric transcription factor HIF‐1 (hypoxia‐inducible factor 1), consisting of a critically regulated HIF‐1α subunit and a constitutively expressed HIF‐1β subunit, is a master regulator of genes involved in adaptation and survival under low‐oxygen conditions. Increased levels of HIF‐1 activity are associated with increased tumor aggressiveness, therapeutic resistance, and mortality.

METHODS

We studied 402 prostate cancer patients for the presence of the 1772C > T (P582S) and 1790G > A (A588T) mutations within the oxygen‐dependent domain of HIF‐1α.

RESULTS

Homozygosity for the P582S mutation was fourfold greater among prostate cancer patients compared to controls (OR = 4.10 [C.I. 95% 1.11 < OR < 17.87], P = 0.018). The existence of this mutation in prostate cancer patients was not associated with any of the clinical or pathological characteristics of the disease. No significant differences were found between the frequencies of A588T mutation in prostate cancer patients and controls.

CONCLUSIONS

Our data suggest that homozygous HIF1A P582S mutation confers significant susceptibility to prostate cancer. Prostate 67:8–13, 2007. © 2006 Wiley‐Liss, Inc.
Castration‐induced epithelial cell death in human prostate tissue is related to locally reduced IGF‐1 levels
Authors: Ohlson Nina , Bergh Anders , Stattin Pär , Wikström Pernilla
Published: September 2006

BACKGROUND

Castration rapidly reduces stroma insulin‐like growth factor (IGF)‐1 synthesis and action in mouse prostate epithelium. We explore if similar changes are of importance for castration‐induced prostate regression in humans.

METHODS

Epithelial and surrounding stroma cells were micro‐dissected from patient biopsies obtained before and shortly after castration. IGF‐1 mRNA levels were quantified by RT‐PCR and related to epithelial apoptosis and IGF‐1, IGF‐1 receptor, and androgen receptor (AR) immunoreactivity.

RESULTS

IGF‐1 mRNA was principally produced in the stroma and IGF‐R1 in the epithelium. Stroma IGF‐1 mRNA levels were significantly decreased after castration in non‐malignant but not malignant tissue. Lack of stroma IGF‐1 reduction after castration was associated with low stroma AR expression before therapy. Reduction of IGF‐1 mRNA levels in the tumor stroma and/or epithelium was associated with epithelial apoptosis after therapy.

CONCLUSIONS

Low AR expression and maintained stroma IGF‐1 synthesis may result in limited tumor cell death after castration therapy. Prostate 67:32–40, 2007. © 2006 Wiley‐Liss, Inc.