bluuebunny/crossref_metadata_2025_split huggingface dataset:
An NMR study of cyclodextrin complexes of the steroidal neuromuscular blocker drug Rocuronium Bromide
Authors: Cameron Kenneth S. , Fletcher Dan , Fielding Lee 
Published: August 2002

AbstractThe interaction of Rocuronium Bromide, and a model steroid Org 7402, with three cyclodextrins (β‐cyclodextrin, γ‐cyclodextrin and Org 25969) was studied by solution state NMR experiments. Stoichiometries and binding constants were determined from 1H chemical shift titrations. All of the systems formed 1 : 1 complexes. Most of the complexes were in fast exchange with unbound species on the NMR time scale, but the most tightly bound complex (Rocuronium Bromide–Org 25969) was in the slow exchange regime. The geometry of the complexes was inferred from 1H and 13C NMR shift changes upon complexation and from intramolecular NOE correlations. Rocuronium Bromide forms a weak complex with β‐cyclodextrin (Ka = 3.3 ± 0.5 × 103 M−1) and no clear picture of the structure of the complex emerges. The complexes with γ‐cyclodextrin (Ka = 1.8 ± 0.2 × 104 M−1) and Org 25969 (Ka > 105 M−1) are true inclusion complexes with the steroid located inside the central void of the cyclodextrin. Copyright © 2002 John Wiley & Sons, Ltd.
1H and 13C NMR spectral assignments for 3,5‐disubstituted tetrahydro‐2H‐ 1,3,5‐thiadiazine‐2‐thione derivatives
Authors: Molero Dolores , Pérez Rolando , Martinez Roberto , Suárez Margarita , Rodriguez Hortensia , Martín Nazario , Seoane Carlos 
Published: August 2002

AbstractThe proton and carbon spectra of 3,5‐disubstituted tetrahydro‐2H‐1,3,5‐thiadiazine‐2‐thione derivatives, prepared by a solid‐phase procedure, are reported. Copyright © 2001 John Wiley & Sons, Ltd.
A 15N NMR investigation of a series of benzotriazinones and related antitumour heterocycles
Authors: Vaughan Keith , Wilman Derry E. V. , Wheelhouse Richard T. , Stevens Malcolm F. G. 
Published: August 2002

AbstractA series of 3‐substituted 1,2,3‐benzotriazin‐4‐ones, 1 and 2, were synthesized by standard methods and the 15N NMR spectra were recorded. All spectra were obtained using the natural abundance of the nitrogen‐15 isotope. The chemical shifts appear in the normal range for N‐1, N‐2 and N‐3 of the triazine ring, and also correlate with the chemical shifts in the spectra of the imidazolotriazinone, 4, and the imidazolotetrazinone, 5. Significantly, the spectra of 1a, 2 and 4, recorded with full NOE, show inversion of the singlet assigned to N‐3, demonstrating that these compounds exist in the tautomeric form shown. The structure of the 4‐iminobenzotriazinone (3) was confirmed by this 15N NMR analysis. The spectrum shows a signal for the NH‐bearing imino‐nitrogen atom, which is an inverted singlet in the NOE spectrum, whereas the signal from the N‐3 atom of 3 is not inverted in the NOE spectrum. Copyright © 2002 John Wiley & Sons, Ltd.
In vivo versus in vitro oviductal glycoprotein (OGP) association with the zona pellucida (ZP) in the hamster and baboon
Authors: O'Day‐Bowman M.B. , Mavrogianis P.A. , Minshall R.D. , Verhage H.G. 
Published: August 2002

AbstractThe goal of this study was to determine if differences exist between in vivo vs. in vitro OGP association with the ZP and to quantitate those differences. Ovarian oocytes were harvested 12.5 or 27 hr post‐hCG from hyperstimulated hamsters or baboons, respectively. Hamster and baboon ovarian oocytes were incubated in vitro in media ± homologous OGP (100 or 200 μg/100 μl) or in some studies with 100 μl oviductal fluid for 3, 6, or 24 hr at 37°C. Some of the baboon ovarian oocytes were transferred immediately after harvesting to the ampulla of both oviducts using a tom cat catheter and retrieved after a 3 hr in situ incubation. Hamster oviductal oocytes were collected 3, 6, and 24 hr following ovulation. After incubation or oocyte retrieval from the oviduct, cumulus cells were removed, oocytes were washed extensively and binding of OGP to the ZP was examined by immunofluorescence. Fluorescence intensity was quantified using densitometric scanning of photographic negatives with the background of each negative as an internal control. In all studies, OGP association with the ZP was significantly greater in vivo than in vitro (P < 0.05). In vitro OGP association with the ZP did not significantly increase with incubation time or OGP concentration; however, a small nonsignificant increase in OGP association with the ZP in the oviduct was detected over time. Differences did not appear to be due to depletion of OGP from the in vitro incubation media, since Western blot analysis of the media showed that OGP was still present. Although OGP concentration in vivo is unknown, Western blots showed similar intensity comparing 100 μg of OGP media and oviductal fluid. Immunolocalization of OGP using laser confocal microscopy showed regional differences in OGP binding. The outer half of the zona pellucida had significantly more OGP bound than the inner half on oviductal oocytes. No regional differences were detected for in vitro incubated oocytes. In conclusion, OGP association with the ZP is greater in vivo vs. in vitro, suggesting that one must be cautious in designing and evaluating in vitro studies of OGP function. Mol. Reprod. Dev. 62: 248–256, 2002. © 2002 Wiley‐Liss, Inc.
Effects of aromatase inhibition on in vitro follicle and oocyte development analyzed by early preantral mouse follicle culture
Authors: Hu Y. , Cortvrindt R. , Smitz J. 
Published: August 2002

AbstractIn vivo studies on folliculogenesis have documented a relation among intrafollicular steroid content, follicle growth, and oocyte development. This study examined how profound changes in androgen/estrogen ratio would affect mouse in vitro follicular development. Arimidex®, a potent follicular aromatase inhibitor was used for this purpose. Early preantral follicles were cultured for 12 days up to the preovulatory stage. Oocyte's meiotic maturation, spindle and chromosome configurations, in vitro fertilization and preimplantation embryo development were evaluated. Compared to controls, Arimidex reduced E2 concentration in follicle culture medium by a factor 1000, and an expected simultaneous accumulation of testosterone was measured in the conditioned medium. Arimidex treatment provoked a dose‐dependent earlier differentiation of the granulosa cells as judged by an earlier antrallike cavity formation and slightly elevated basal progesterone secretion. Follicle survival exceeded 98% in all groups and all follicles responded normally to HCG/EGF addition on day 12 by cumulus mucification. By the HCG ovulatory challenge, progesterone output was reduced in Arimidex supplemented groups suggesting preovulatory luteinization. These results indicate that in vitro mouse follicles can develop normally under very low levels of estrogens and that a local androgen increase by a factor 3 is not atretogenic. Oocyte growth did not differ among culture conditions. Arimidex treatment induced a dose dependent enhancement of GVBD and polar body formation rate in response to HCG at the end of culture. Spindle and chromosome analyses demonstrated that in all groups, 90% of the oocytes which extruded a polar body had also reached the MII stage. While most of the cultured MII oocytes had a normal spindle and well aligned chromosomes, significantly less oocytes were fertilized in the groups cultured in the presence of Arimidex. Once fertilized, however, there was found to be no difference for preimplantation embryo development between controls and Arimidex treatment. These data suggest that in mice a pronounced estrogenic environment is not essential for in vitro folliculogenesis. Drastic changes in the intrafollicular steroid concentrations do not disrupt meiotic maturation nor compromise early preimplantation development, but adversely affect fertilization of in vitro grown oocytes. Mol. Reprod. Dev. 61:549–559, 2002. © 2002 Wiley‐Liss, Inc.
Expression of p53 in luminal and glandular epithelium during the growth and regression of rat uterus during the estrous cycle
Authors: Mendoza‐Rodríguez Carmen Adriana , Merchant‐Larios Horacio , Segura‐Valdez María de Lourdes , Moreno‐Mendoza Norma , Cruz María Esther , Arteaga‐López Paola , Camacho‐Arroyo Ignacio , Dominguez Roberto , Cerbón Marco 
Published: August 2002

AbstractIt has been well recognized that epithelial cells of the rat endometrium cyclically proliferate and die during the estrous cycle. The aim of the present study was to determine p53 expression pattern and correlate it with the the apoptotic pattern of epithelial cells of the rat uterus during the estrous cycle. The p53 mRNA and protein expression pattern was assessed by in situ hybridization and immunohistochemistry. The apoptotic index was determined by using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and electron microscopy. The highest p53 mRNA content, detected by in situ hybridization, was observed on the metestrus day both in the luminal and the glandular epithelia. During this period both epithelia presented high proliferation. The content of p53 mRNA markedly decreased in the following days, presenting its minimal values on the estrus day. The highest number of p53 immunopositive nuclei, in both the luminal and the glandular epithelia, was also detected on the metestrus day, while the lowest one was found on estrus day. On the proestrus day, p53 protein was predominantly detected in the glandular epithelium. However, on the estrus day, p53 protein was detected both in the nuclei and in the cytoplasm of luminal epithelial cells, predominantly in the cytoplasm. The highest apoptotic index in both the luminal and the glandular epithelia was observed on the estrus day whereas the lowest one was observed on the proestrus day. The apoptotic index values were higher in the luminal than in the glandular epithelia. The overall results indicate that p53 expression at both mRNA and protein levels is higher on the metestrus day when the apoptotic index is low. This suggests that p53 should play an important physiological role during proliferative phases of the estrous cycle in the rat uterus. Mol. Reprod. Dev. 61:445–452, 2002. © 2002 Wiley‐Liss, Inc.
Production of germ‐line chimeras in rainbow trout by blastomere transplantation
Authors: Takeuchi Yutaka , Yoshizaki Goro , Takeuchi Toshio 
Published: August 2002

AbstractWe describe a technique for producing germ‐line chimeric rainbow trout, Oncorhynchus mykiss, by microinjection of the isolated blastomeres. FITC‐labeled donor cells and non‐labeled recipient embryos at various developmental stages between the early blastula and early gastrula stages were used for cell transplantation. The chimera formation rate and the degree of donor cell distribution in recipient embryos were evaluated at both the late gastrula stage (5 days post fertilization (dpf)) and the 40‐somite stage (10 dpf). Among the six combinations of developmental stages of donor and recipient embryos, the combination of midblastula (2.5 dpf) donor cells and early blastula (1.5 dpf) recipient embryos gave the highest chimera formation rate and the best distribution pattern of donor cells. Using this combination, chimeric rainbow trout were produced with donor blastomeres from dominant orange‐colored mutant embryos and wild‐type recipient embryos. Of the 238 chimeric embryos produced, 28 (12%) hatched normally and 14 of the 28 fry (50%) had donor‐derived orange body color. To test for germ‐line transmission of donor cells, gametes obtained from the matured chimeras were fertilized with gametes from wild‐type fish. Of the 19 matured chimeras, 6 (32%) yielded donor‐derived orange‐colored progeny, in addition to wild‐type siblings. The contribution rates of donor cells in the germ‐line ranged from 0.3 to 14%. This technique for producing germ‐line chimeras should be a powerful tool for cell‐mediated gene transfer in rainbow trout. Especially, if body color mutants are used for either donor cells or the host embryos, it will be possible to easily concentrate F1 transgenic embryos derived from transplanted donor cells by body color screening. Mol. Reprod. Dev. 59: 380–389, 2001. © 2001 Wiley‐Liss, Inc.
Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing
Authors: Chatterjee Suvro , Gagnon Claude 
Published: August 2002

AbstractIn the present study, we provide evidence for the production of reactive oxygen species (ROS) during cryopreservation of bovine spermatozoa. Cooling and thawing of spermatozoa cause an increase in the generation of superoxide radicals. Although nitric oxide production remains unaltered during sperm cooling from 22–4°C, a sudden burst of nitric oxide radicals is observed during thawing. Increase in lipid peroxidation levels have been observed in frozen/thawed spermatozoa and appears to be associated with a reduction in sperm membrane fluidity as detected by spin labeling studies. The data presented provide strong evidence that oxygen free radicals are produced during freezing and thawing of bovine spermatozoa and suggest that these reactive oxygen species may be a cause for the decrease in sperm function following cryopreservation. Mol. Reprod. Dev. 59: 451–458, 2001. © 2001 Wiley‐Liss, Inc.
Family of SRY/Sox proteins is involved in the regulation of the mouse Msh4 (MutS homolog 4) gene expression*
Authors: Santucci‐Darmanin Sabine , Vidal Frédérique , Scimeca Jean‐Claude , Turc‐Carel Claude , Paquis‐Flucklinger Véronique 
Published: August 2002

AbstractThe eukaryotic MutHLS‐like system plays a crucial role in both mitosis and meiosis. Until now, a number of works have focused on the function of MutS and MutL homologs during mismatch DNA repair. Nevertheless, little is known about the role of these proteins during meiosis. MSH4 is a meiosis specific protein that is necessary for meiotic recombination in Saccharomyces cerevisiae. The human MSH4 protein is only found in testis and ovary. It is involved first in synapsis and second during recombination together with MLH1 (MutL Homolog 1). Here, we report the identification of the mouse Msh4 gene that is located on chromosome 3. We examined the expression of mMsh4 in testes of increasing developmental age and in elutriated germ cells. The pattern of expression during spermatogenesis is consistent with a role for MSH4 both during zygonema and pachynema. We demonstrated a promoter activity of the mMsh4 5′‐flanking region by cell transfection experiments with a luciferase reporter gene. We found several SRY/Sox binding sites in this region and co‐transfection experiments showed that SRY could down regulate mMsh4 promoter transcriptional activity. We propose that the regulation of mMsh4 expression could be one of the reasons for the persistence of SRY and/or SRY‐related proteins in adult testis. Mol. Reprod. Dev. 60: 172–180, 2001. © 2001 Wiley‐Liss, Inc.
Degradation of maternal cdc25c during the maternal to zygotic transition is dependent upon embryonic transcription*
Authors: Anderson Jon E. , Matteri Robert L. , Abeydeera Lalantha R. , Day Billy N. , Prather Randall S. 
Published: August 2002

AbstractTo gain a better understanding of the molecular differences that may contribute to cleavage arrest and the poorer development associated with laboratory produced embryos, a series of experiments were conducted to quantitate the message levels of the cell cycle controller cdc25c, over the maternal to zygotic transition (MZT) in 4‐cell in vivo‐ and in vitro‐derived porcine embryos. The experiments were designed to measure both maternal and embryonic derived cdc25c transcripts by quantitative reverse transcription‐competitive polymerase chain reaction (RT‐cPCR), determine the point of the transition to zygotic genome activation, and study the interaction between initial embryonic transcription and maternal cdc25c degradation. Analysis of in vivo‐ and in vitro‐derived embryos revealed no difference in cdc25c message level for any of the times P4CC (P > 0.05). Comparison of control embryos from 5‐ to 33‐hr P4CC revealed a reduction in transcript quantities in the 10‐hr P4CC group that was maintained at later time points (P < 0.05). Embryos cultured in the RNA polymerase inhibitor, α‐amanitin, from cleavage to 5‐, 10‐, 18‐, 25‐, or 33‐hr P4CC displayed no difference in cdc25c levels when compared to controls at similar time points (P > 0.05). However, if embryos were first exposed to α‐amanitin after 18‐hr P4CC with this treatment continuing to 33 hr, the levels of cdc25c transcript were reduced (P < 0.04) when compared to those embryos that were first exposed to the inhibitor at either 5‐ or 10‐hr P4CC. This finding and the comparison of these same embryos to the 0–33‐hr α‐amanitin and control groups allowed us to conclude that cdc25c transcription began between 10‐ and 18‐hr P4CC, with the degradation of maternal cdc25c message dependent on transcriptional initiation. Mol. Reprod. Dev. 60: 181–188, 2001. © 2001 Wiley‐Liss, Inc.
Production of transgenic mice from vitrified pronuclear‐stage embryos
Authors: Bagis H. , Odaman H. , Sagirkaya H. , Dinnyés A. 
Published: August 2002

AbstractCryopreservation of pronuclear‐stage embryos would be useful for transgenic technology and genome preservation purposes. We compared a novel vitrification technique (solid surface vitrification, SSV) with another vitrification method in straws for cryosurvival and to generate transgenic progeny from cryopreserved mouse zygotes following microinjection. The SSV solution consisted of 35% ethylene glycol (EG), 5% polyvinyl‐pyrrolidone (PVP), and 0.4 M trehalose in M2 supplemented with 4 mg/ml BSA; the in straw vitrification solution was 7 M EG in M2 plus BSA. In experiment I, we compared the effect of the vitrification solutions alone, without cooling. No reduction was detected in survival and cleavage rates. In experiment II, SSV yielded a significantly higher percentage of morphologically normal zygotes (96%) that also cleaved at significantly higher rates (80%) when compared to that following “in straw” vitrification (68 and 66%, respectively). Cleavage rate in the non‐vitrified control group (93%) was significantly higher than that of both vitrified groups. Following embryo transfer, there was no difference in the rate of pups obtained from the SSV, “in straw” vitrified, and control groups (97/457, 21%; 15/75, 20% and 56/209, 27%, respectively). In experiment III, SSV vitrified and fresh embryos were used for pronuclear DNA injection. Survival rate of vitrified embryos after microinjection was reduced compared to nonvitrified ones (64 vs. 72%, respectively; P < 0.05); however, development to two‐cell stage was not different (76 vs. 72%, respectively). Following embryo transfer of vitrified vs. fresh microinjected embryos, in both cases 10% live pups were generated, including transgenic pups. The results demonstrated that the efficiency of generating transgenic pups from SSV vitrified pronuclear zygotes is comparable to that from fresh embryos. Mol. Reprod. Dev. 61: 173–179, 2002. © 2002 Wiley‐Liss, Inc.
Down syndrome and Alzheimer's disease: A link between development and aging
Authors: Lott Ira T. , Head Elizabeth 
Published: August 2002

AbstractA subset of aged individuals with Down syndrome (DS) exhibits the clinical features of Alzheimer's disease (AD) but our ability to detect dementia in this population is hampered by developmental differences as well as the sensitivity of existing test tools. Despite the apparent clinical heterogeneity in aged individuals with DS, age‐associated neuropathology is a consistent feature. This is due to the fact that trisomy 21 leads to a dose‐dependent increase in the production of the amyloid precursor protein and subsequently the production of the amyloidogenic fragments leading to early and predominant senile plaque formation. A review of the existing literature indicates that oxidative damage and neuroinflammation may interact to accelerate the disease process particularly in individuals with DS over the age of 40 years. By combining clinical information with measures of brain‐region specific neuropathology we can “work backwards” and identify the earliest and most sensitive clinical change that may signal the onset of AD. For the past 50 years, investigators in the fields of mental retardation, developmental disabilities, and aging have been interested in the curious link between AD and DS. The morphologic and biochemical origins of AD are seen in the early years of the lifespan for individuals with DS. Study of the process by which AD evolves in DS affords an opportunity to understand an important link between development and aging. This review will focus on advances in the molecular and clinical basis of this association. MRDD Research Reviews 2001;7:172–178. © 2001 Wiley‐Liss, Inc.
Compartmental study of T1 and T2 in rat brain and trigeminal nerve in vivo
Authors: Does Mark D. , Gore John C. 
Published: August 2002

AbstractThe integrated T1–T2 characteristics of rat brain and trigeminal nerve water were studied in vivo using a rapid method for acquiring a series of images that depend on T1 and T2 relaxation times. Gray matter regions showed only one signal component in both the T1 and T2 domains. Trigeminal nerve, however, which has been shown previously to exhibit three T2 components, was found to also exhibit three T1 components. The correlations between these T1 and T2 components were demonstrated by uniquely filtering out each of the three T2 components using an inversion‐recovery preparation, as determined by the component T1 values. Based on previous works, it is postulated that each of these three signal components is derived from a unique microanatomical region of the nerve. Knowledge of these T1 components may thus prove valuable in devising novel methods of identifying the presence and quantifying the volume of tissue subtypes such as myelin. Magn Reson Med 47:274–283, 2002. © 2002 Wiley‐Liss, Inc.
Recovery of function in cytoprotected cerebral cortex in rat stroke model assessed by functional MRI
Authors: Sauter Andre , Reese Torsten , Pórszász Robert , Baumann Diana , Rausch Martin , Rudin Markus 
Published: August 2002

AbstractFunctional recovery in cytoprotected somatosensory cortex in a rat stroke model was studied using functional MRI (fMRI). Calcium antagonist treatment (isradipine) following permanent middle cerebral artery occlusion (pMCAO) reduced the infarct volume by 33 ± 9%. The somatosensory cortex representing the forepaws was spared from infarction; however, cerebral blood flow (CBF) was significantly reduced in this area 24 hr following pMCAO. Neural function was assessed at days 1, 2, 5, and 12 following pMCAO by fMRI using electrical stimulation of both forepaws. Vehicle‐treated rats did not show fMRI responses in the infarcted somatosensory cortex throughout the study. Several of the isradipine‐treated animals displayed functional recovery in the cytoprotected cortex at days 5 (3/5 rats) and 12 (5/10). Correlations with fMRI signals showed that normal T2 and ADC values in the respective brain areas are necessary, but not sufficient prerequisites for functionality. Recovery of neural function is associated with normalization of CBF in the cytoprotected brain area. Magn Reson Med 47:759–765, 2002. © 2002 Wiley‐Liss, Inc.