bluuebunny/crossref_metadata_2025 huggingface dataset:
Hereditary hemorrhagic telangiectasis treated by the harmonic scalpel
Authors: Ishibashi Toshio , Takamatsu Shunsuke
Published: March 2003

Background.

Hereditary hemorrhagic telangiectasis (HTT) is a familial autosomal dominant genetic disorder that causes abnormalities of the wall of peripheral blood vessels. Severe nosebleed often is the dominant symptom. A variety of therapies have been proposed for epistaxis control in HHT but with limited success.

Methods.

We report two cases of HHT in which recurrent nasal bleeding was successfully controlled using the Harmonic Scalpel.

Results.

Use of the Harmonic Scalpel avoids the carbonization and incrustation of the nasal mucosa that commonly results from electrocautery and laser irradiation. Even during active bleeding, hemostasis can be achieved by repeated applications of the scalpel blade.

Conclusion.

This method can be performed quick and reliably on an outpatient basis. The Harmonic Scalpel may be the treatment of choice for recurrent epistaxis in HHT. © 2003 Wiley Periodicals, Inc. Head Neck 25: 333–336, 2003
The impact of head and neck cancer and facial disfigurement on the quality of life of patients and their partners
Authors: Vickery Lindsey E. , Latchford Gary , Hewison Jenny , Bellew Maggie , Feber Tricia
Published: March 2003

Background.

Psychological and physical stresses from head and neck cancer can be substantial for patients and partners. There is minimal research exploring treatment impact, particularly facial disfigurement after surgery.

Materials and Methods.

Twenty‐eight surgery and radiotherapy/brachytherapy/chemoradiation patients and 25 of their partners were compared with 23 radiotherapy/brachytherapy patients and 19 partners. Participants completed the Hospital Anxiety and Depression Scale, Psychosocial Adaptation to Illness Scale, Dyadic Adjustment Scale, and European Organisation for Research and Treatment of Cancer Quality Of Life Scale, including the Head and Neck Cancer module. The Dropkin Disfigurement and Dysfunction scale classified surgical impairment.

Results.

Partners reported greater distress than patients on some scales. Patients did not have a lower quality of life compared with normal populations and other cancer patients. However, on the EORTC they did and were comparable to a normed sample of head and neck cancer patients. Treatment modality was not predictive of psychological vulnerability.

Discussion.

Head and neck cancer patients do not necessarily experience poor quality of life. The disease can have a significant impact on partners. © 2003 Wiley Periodicals, Inc. Head Neck 25: 289–296, 2003
Squamous cell carcinoma of the buccal mucosa: One institution's experience with 119 previously untreated patients
Authors: Diaz Eduardo M. , Holsinger F. Christopher , Zuniga Edgar R. , Roberts Dianna B. , Sorensen Douglas M.
Published: March 2003

Background.

Squamous cell carcinoma (SCC) of the buccal mucosa is a rare, but especially aggressive, form of oral cavity cancer, associated with a high rate of locoregional recurrence and poor survival. We reviewed our institution's experience with 119 consecutive, previously untreated patients with buccal SCC.

Methods.

We reviewed the charts of 250 patients who were seen at The University of Texas M. D. Anderson Cancer Center between January, 1974, and December, 1993. Of these, 119 were untreated and were subsequently treated exclusively at our institution. Patients who were previously treated elsewhere or whose lesions arose in other sites and only secondarily involved the buccal mucosa were excluded.

Results.

Patients with T1‐ or T2‐sized tumors had only a 78% and 66% 5‐year survival, respectively. Muscle invasion, Stensen's duct involvement, and extracapsular spread of involved lymph nodes were all associated with decreased survival (p < .05). Surgical salvage for patients with locoregional recurrence after radiation therapy was rarely successful.

Conclusions.

SCC of the buccal mucosa is a highly aggressive form of oral cavity cancer, with a tendency to recur locoregionally. Patients with buccal mucosa SCC have a worse stage‐for‐stage survival rate than do patients with other oral cavity sites. © 2003 Wiley Periodicals, Inc. Head Neck 25: 267–273, 2003
Functional anatomy of the lymphatic drainage system of the upper aerodigestive tract and its role in metastasis of squamous cell carcinoma
Authors: Werner Jochen A. , Dünne Anja A. , Myers Jeffrey N.
Published: March 2003

Background.

Although there is a significant understanding of the vascular anatomy of the upper aerodigestive tract (UADT), there is less detailed knowledge of the architecture and drainage patterns of the lymphatic system. Detailed knowledge of the lymphatic system is critical for understanding the role of sentinal node identification in the management of different cancers.

Methods.

We have combined microscopic techniques with in vivo and in vitro lymphographic studies to survey the architecture and drainage patterns of the lymphatic system of the UADT in 850 organ specimens.

Results.

These studies show an interaction of superficial and deep lymphatic networks that vary in density but have a constant distribution characterized by predictable patterns of lymph drainage into the regional lymph nodes.

Conclusions.

Detailed knowledge of the lymphatic system of the UADT contributes to a better understanding of the patterns of metastatic spread of carcinomas of the UADT and provides a strong rationale for the practice of sentinel node identification in the management of these tumors. © 2003 Wiley Periodicals, Inc. Head Neck 25: 322–332, 2003
Validation of a non‐invasive blood‐sampling technique for doubly‐labelled water experiments
Authors: Voigt Christian C. , Helversen Otto Von , Michener Robert H. , Kunz Thomas H.
Published: March 2003

Two techniques for bleeding small mammals have been used in doubly‐labeled water (DLW) studies, including vena puncture and the use of starved nymphal stages of hematophagous reduviid bugs (Reduviidae, Hemiptera). In this study, we tested the validity of using reduviid bugs in doubly‐labeled water experiments. We found that the isotope enrichment in initial blood samples collected with bugs was significantly lower compared to isotope enrichment in blood samples obtained using vena puncture. We therefore used the desiccation method for estimating total body water (TBW) in DLW experiments because TBW calculated using the isotope dilution method was overestimated when blood samples were collected using reduviid bugs. In our validation experiment with nectar‐feeding bats (Glossophaga soricina), we compared estimates of daily energy expenditure (DEE) using DLW with those derived from the energy balance method. We considered Speakman's equation (controlling for 25% fractionated water loss) as the most appropriate for our study animal and calculated DEE accordingly. On average, DEE estimated with DLW was not significantly different from the mean value obtained with the energy balance method (mean deviation 1.2%). We conclude that although bug hemolymph or intestinal liquids most likely contaminate the samples, estimates of DEE are still valid because the DLW method does not depend on absolute isotope enrichments but on the rate of isotope decrease over time. However, dilution of blood with intestinal liquids or hemolymph from a bug may lead to larger variation in DEE estimates. We also tested how the relative error of DLW estimates changed with varying assumptions about fractionation. We used three additional equations for calculating DEE in DLW experiments. The basic equation for DLW experiments published by Lifson and McClintock (LM‐6) assumes no fractionation, resulted in an overestimate of DEE by 10%. Nagy's equation (N‐2) controls for changes in body mass but not for fractionation. Using Nagy's equation, DEE was overestimated by 8%. Under the assumption that 50% of total water flux fractionates, the alternative equation by Lifson and McClintock (LM‐35) DEE was underestimated by 5%. The best fit between estimates of DEE based on DLW and energy balance measurements was derived by assuming that 32% of total water flux (TWF) is fractionated. We conclude that the outcome of DLW experiments is sensitive to assumptions regarding evaporative water loss, and thus recommend Speakman's equation 7.17 for use with bats. J. Exp. Zool. 296A:87–97, 2003. © 2003 Wiley‐Liss, Inc.
Differential effects of thyroxine on metabolic enzymes and other macromolecules in a freshwater teleost
Authors: Tripathi G. , Verma Priyanka
Published: March 2003

The effects of thyroxine (T4) on citrate synthase (CS), glucose 6‐phosphate dehydrogenase (G6‐PDH), lactate dehydrogenase (LDH), DNA, RNA, and protein of various tissues were studied to elucidate the hormonal control of metabolism in a freshwater catfish, Clarias batrachus. T4 did not produce any significant effect on DNA content of the fish. The CS, RNA, and protein contents of brain, liver, and skeletal muscle of the fish exposed to thiourea for 28 days decreased approximately 50–58% as compared to their levels in control individuals. Injection of T4 to thiourea‐exposed fish produced about three‐fold increases in CS, RNA, and protein. These macromolecular inductions by T4 were blocked by actinomycin D or cycloheximide. This suggests T4‐induced de novo synthesis of macromolecules and stimulation of aerobic capacity. However, the activities of G6‐PDH and LDH of brain, liver, and skeletal muscle of the fish exposed to thiourea increased two times that of the activities in control individuals. Administration of T4 to thiourea‐exposed fish reduced LDH and G6‐PDH activities by about 64–74%, which reflects T4‐dependent inhibition in anaerobic power and selective anabolic activities of the HMP pathway. These differential effects of T4 on some metabolic enzymes and other important macromolecules may be to meet the other T4‐induced responses in the freshwater catfish. J. Exp. Zool. 296A:117–124, 2003. © 2003 Wiley‐Liss, Inc.
Photoperiodic response of the hypothalamo‐pituitary‐gonad axis in male and female canaries, Serinus canaria
Authors: Bentley George E. , Audage Natalie C. , Hanspal Era K. , Ball Gregory F. , Hahn Thomas P.
Published: March 2003

Although the gonadal photoperiodic response and its influence upon the song control system in canaries have been extensively studied, photoperiodic regulation of the GnRH system has not been investigated. To examine the relationship between photoperiod and the reproductive neuroendocrine system in male and female canaries, three groups of canaries were exposed to chronic short days (8L:16D; Phsens), acute long days (18L:6D; Phstim) and chronic long days (also 18L:6D; Phrefr) to induce the reproductive states of photosensitivity, photostimulation, and photorefractoriness, respectively. Brain sections were processed for GnRH immunocytochemistry. The canaries in this study did not demonstrate consistent or uniform responses to different photoperiodic treatments. In males, gonad size varied with photoperiod; Phstim males had larger gonads than either the Phsens or Phrefr males. In contrast, there was no difference between groups in female gonad size as a result of photoperiodic treatment. Brain GnRH cell number, cell size, and fiber density were similar in all groups. The results suggest that canaries are not as obligatory photoperiodic as previously thought (or at least not all varieties of domestic canaries are). This could be a result of many years of domestication, the natural history of the species, phylogenetic constraint, or a combination of these factors. J. Exp. Zool. 296A:143–151, 2003. © 2003 Wiley‐Liss, Inc.
Changes in localization of immune cells and cytokines in corpora lutea during luteolysis in murine ovaries
Authors: Komatsu Kohji , Manabe Noboru , Kiso Minako , Shimabe Munetake , Miyamoto Hajime
Published: March 2003

Immune cells, which constitute a significant cell mass in the corpora lutea (CLs), are considered to play critical roles in luteolysis, but the details are not fully understood. We histochemically investigated the changes in distribution and cell density of macrophages and T lymphocytes and in tumor necrosis factor (TNF)‐α and interferon (IFN)‐γ, which can induce apoptosis in the luteal cells in murine CLs during luteal regression. No macrophages or T lymphocytes were observed in functionally healthy CLs. Abundant macrophages and increasing T lymphocytes were demonstrated in CLs at the functional regression stage (early stage of regression). At the structural regression stage (late stage of regression), abundant T lymphocytes but no macrophages were demonstrated in the CLs. A moderate amount of TNF‐α was detected in all CLs at all stages. No IFN‐γ was detected in either healthy or early regressing CLs, but a large amount of IFN‐γ was detected at the late regression stage. Moreover, in cultured luteal cells, reactivity against Fas‐ligand (FasL) was caused by pretreatment with TNF‐α and IFN‐γ and apoptosis was induced by FasL treatment. These findings support the hypothesis that macrophages initiate T lymphocyte aggregation at the early stage of luteal regression, and then T lymphocytes induce apoptosis on luteal cells, which in turn develop sensitivity against FasL by TNF‐α and IFN‐γ. J. Exp. Zool. 296A:152–159, 2003. © 2003 Wiley‐Liss, Inc.
Expression of heat shock and cold shock proteins in the gorgonian Leptogorgia virgulata
Authors: Kingsley Roni J. , Afif Evelyn , Cox Brandon C. , Kothari Sonali , Kriechbaum Karmon , Kuchinsky Katherine , Neill Anna T. , Puri Anita F. , Kish Valerie M.
Published: March 2003

In this study, we analyzed the response of the temperate, shallow‐water gorgonian, Leptogorgia virgulata, to temperature stress. Proteins were pulse labeled with 35S‐methionine/cysteine for 1 h to 2 h at 22°C (control), or 38°C, or for 4 h at 12.5°C. Heat shock induced synthesis of unique proteins of 112, 89, and 74 kDa, with 102, 98 and 56 kDa proteins present in the control as well. Cold shock from 22°C–12.5°C induced the synthesis of a 25 kDa protein, with a 44 kDa protein present in the control as well. Control samples expressed unique proteins of 38, and 33 kDa. Non‐radioactive proteins expressed under the same conditions as above, as well as natural field conditions, were tested for reactivity with antibodies to heat shock proteins (HSPs). HSP60 was the major protein found in L. virgulata. Although HSP47, HSP60, and HSP104 were present in all samples, the expression of HSP60 was enhanced in heat stressed colonies, while HSP47 and HSP104 expression were greatest in cold shocked samples. Inducible HSP70 was expressed in cold‐shocked, heat‐shocked, and field samples. Constitutively expressed HSP70 was absent from all samples. The expression of HSP90 was limited to heat shocked colonies. The expression of both HSP70 and HSP104 suggests that the organism may also develop a stress tolerance response. J. Exp. Zool. 296A:98–107, 2003. © 2003 Wiley‐Liss, Inc.
Evolvability and genetic constraint in Dalechampia blossoms: Genetic correlations and conditional evolvability
Authors: Hansen Thomas F. , armbruster W. Scott , Carlson Matthew L. , PÉlabon Christophe
Published: March 2003

The short‐term evolvability of a character is closely related to its level of additive genetic variation. However, a large component of the variation in any one character may be pleiotropically linked to other characters under the influence of different selective factors. Therefore, the organization of the organism into quasi‐independent modules may be an important prerequisite for evolvability. In this paper we propose to study character evolvability in terms of conditional genetic variation. By estimating the amount of genetic variation in a character, y, that is independent of other characters, x, we can assess the evolvability of y when there is stabilizing selection on x. We suggest that systematic use of conditioning may help build a picture of modular organization and quasi‐independent evolvability. As an illustration, we use this approach to assess the evolvability of floral characters in Dalechampia scandens (Euphorbiaceae). Although our study population had relatively low levels of genetic variation at the outset, we find evidence that conditioning may lead to substantial further reduction in the genetic variation available for independent adaptation. This provides additional evidence that the D. scandens blossom is constrained in its short‐term evolvability. J. Exp. Zool. (Mol. Dev. Evol.) 296B:23–39, 2003. © 2003 Wiley‐Liss, Inc.
Embryological origins of developmental stability: Size, shape and fluctuating asymmetry in prenatal random bred mice
Authors: Hallgrímsson Benedikt , Miyake Tsutomo , Wilmore Katherine , Hall Brian K.
Published: March 2003

Ontogenetic patterns of fluctuating asymmetry (FA) can be used to test models for the mechanisms underlying stability during embryonic development (developmental stability). In this study, we ask whether developmental processes initially show high levels of instability that are subsequently dampened through active compensatory mechanisms or passive properties of developmental systems or whether the effects of instability accumulate during embryonic development causing random drift away from an earlier stable state. Previous work on this question has dealt with postnatal skeletal growth and thus been unable to effectively distinguish developmental instability from the effects of mechanically mediated variation in bone modeling and remodeling. Here, we report that FA variances of limb skeletal elements in CD1 mice decrease with gestational age from day 14 to birth (day 20.5). Thus, in mouse limbs, skeletal development is characterized by a high level of developmental instability initially that is reduced during subsequent prenatal development. These results are consistent with the existence of active mechanisms that compensate for the effects of minor perturbations or deviations during development. However, they are also consistent with Soule's model of allomeric variation in which the variance of structures is reduced as the number of independent developmental events that produce them increases. This study illustrates that predictions based on morphometric analyses can yield insights into general properties of developmental systems in cases where specific developmental mechanisms are not yet known. J. Exp. Zool. (Mol. Dev. Evol.) 296B:40–57, 2003. © 2003 Wiley‐Liss, Inc.
Pleiotropic effects on mandibular morphology I. developmental morphological integration and differential dominance
Authors: Ehrich Thomas H. , Vaughn Ty T. , Koreishi Safina F. , Linsey Robin B. , Pletscher L. Susan , Cheverud James M.
Published: March 2003

Pleiotropy refers to a single genetic locus that affects more than one phenotypic trait. Pleiotropic effects of genetic loci are thought to play an important role in evolution, reflecting functional and developmental relationships among phenotypes. In a previous study, we examined pleiotropic effects displayed by quantitative trait loci (QTLs) on murine mandibular morphology in relation to mandibular structure and function. In replicating most of our previous QTLs and increasing our sample size, this study strengthens and extends our earlier results. As in our previous study, we find that QTL effects tend to be restricted to developmentally or functionally related traits. In addition, we examine patterns of differential dominance for pleiotropic QTL effects. Differential dominance occurs when dominance patterns for a single locus vary among traits. We find that multivariate overdominance is a common and substantial phenomenon, and may potentially provide an explanation for the persistence of heterozygosity in natural populations. J. Exp. Zool. (Mol. Dev. Evol.) 296B:58–79, 2003. © 2003 Wiley‐Liss, Inc.
Planar embeddings with infinite faces
Authors: Bonnington C. Paul , Watkins Mark E.
Published: March 2003

We investigate vertex‐transitive graphs that admit planar embeddings having infinite faces, i.e., faces whose boundary is a double ray. In the case of graphs with connectivity exactly 2, we present examples wherein no face is finite. In particular, the planar embeddings of the Cartesian product of the r‐valent tree with K2 are comprehensively studied and enumerated, as are the automorphisms of the resulting maps, and it is shown for r = 3 that no vertex‐transitive group of graph automorphisms is extendable to a group of homeomorphisms of the plane. We present all known families of infinite, locally finite, vertex‐transitive graphs of connectivity 3 and an infinite family of 4‐connected graphs that admit planar embeddings wherein each vertex is incident with an infinite face. © 2003 Wiley Periodicals, Inc. J Graph Theory 42: 257–275, 2003
Small degree out‐branchings
Authors: Bang‐Jensen Jørgen , Thomassé Stéphan , Yeo Anders
Published: March 2003

Using a suitable orientation, we give a short proof of a strengthening of a result of Czumaj and Strothmann 4: Every 2‐edge‐connected graph G contains a spanning tree T with the property that $d_T(v)\le {d_T(v)+\,3 \over 2}$ for every vertex v. As an analogue of this result in the directed case, we prove that every 2‐arc‐strong digraph D has an out‐branching B such that $d^+_B(x)\le {d^+_D(x)+ \over 2}+1$. A corollary of this is that every k‐arc‐strong digraph D has an out‐branching B such that $d^+_B(v)\le {d^+_D(v)+ \over 2^r}+r$, where $r=\lfloor \log_2k\rfloor$. We conjecture that in this case $d^+_B(x)\le {d^+_D(x)+ \over k}+1$ would be the right (and best possible) answer. If true, this would again imply a strengthening of a result from 4 concerning spanning trees with small degrees in k‐connected graphs when k ≥ 2. We prove that for acyclic digraphs the existence of an out‐branching satisfying prescribed bounds on the out‐degrees of each vertex can be checked in polynomial time. A corollary of this is that the existence of arc‐disjoint branchings $F^+_s$,$F^-_t$, where the first is an out‐branching rooted at s and the second an in‐branching rooted at t, can be checked in polynomial time for the class of acyclic digraphs © 2003 Wiley Periodicals, Inc. J Graph Theory 42: 297–307, 2003
Eulerian subgraphs in 3‐edge‐connected graphs and Hamiltonian line graphs
Authors: Chen Zhi‐Hong , Lai Hong‐Jian , Li Xiangwen , Li Deying , Mao Jinzhong
Published: March 2003

In this paper, we show that if G is a 3‐edge‐connected graph with $S \subseteq V(G)$ and $|S| \le 12$, then either G has an Eulerian subgraph H such that $S \subseteq V(H)$, or G can be contracted to the Petersen graph in such a way that the preimage of each vertex of the Petersen graph contains at least one vertex in S. If G is a 3‐edge‐connected planar graph, then for any $|S|\le 23$, G has an Eulerian subgraph H such that $S\subseteq V(H)$. As an application, we obtain a new result on Hamiltonian line graphs. © 2003 Wiley Periodicals, Inc. J Graph Theory 42: 308–319, 2003
An algebraic characterization of projective‐planar graphs
Authors: Abrams Lowell , Slilaty Daniel C.
Published: March 2003

We give a detailed algebraic characterization of when a graph G can be imbedded in the projective plane. The characterization is in terms of the existence of a dual graph G* on the same edge set as G, which satisfies algebraic conditions inspired by homology groups and intersection products in homology groups. © 2003 Wiley Periodicals, Inc. J Graph Theory 42: 320–331, 2003
Crossing numbers of sequences of graphs II: Planar tiles
Authors: Pinontoan Benny , Richter R. Bruce
Published: March 2003

We describe a method of creating an infinite family of crossing‐critical graphs from a single small planar map, the tile, by gluing together many copies of the tile together in a circular fashion. This method yields all known infinite families of k‐crossing‐critical graphs. Furthermore, the method yields new infinite families, which extend from (4,6) to (3.5,6) the interval of rationals r for which there is, for some k, an infinite sequence of k‐crossing‐critical graphs all having average degree r. © 2003 Wiley Periodicals, Inc. J Graph Theory 42: 332–341, 2003
Generating cycles in graphs with at most one end
Authors: Bruhn Henning
Published: March 2003

Answering a question of Halin, we prove that in a 3‐connected graph with at most one end the cycle space is generated by induced non‐separating cycles. © 2003 Wiley Periodicals, Inc. J Graph Theory 42: 342–349, 2003
ADAM family genes testase 2α and 2β are chromosomally linked and simultaneously expressed in male germ cells
Authors: Bolcun Ewelina , Rzymski Tomasz , Nayernia Karim , Engel Wolfgang
Published: March 2003

The ADAM (a disintegrin and a metalloprotease) family includes the best‐characterized proteins involved in gamete interaction and membrane fusion in mammals. Previous studies have shown that the murine testase 2 (ADAM 25) gene is expressed specifically in testis. We found two different restriction patterns of subcloned fragments of the gene, indicating the presence of two different testase 2 transcripts. Further experiments and Celera database searches demonstrated that the two transcripts are the products of two testase 2 genes, which are located on mouse chromosome 8 in close distance of 24 kb. They show high sequence similarity to the published testase 2 gene (87.8 and 95.5%, respectively). The genomic structure of both testase 2 genes (α and β) is different from other ADAM family genes like that for cyritestin and fertilin. While these genes are composed of about 20 exons, the testase 2α and 2β genes contain only two exons. The first exon is very short ∼85 bp while the second exon is ∼2.4 kb long. Both testase 2 genes are specifically expressed in testis and they exhibit the same temporal and spatial expression pattern during male germ cell differentiation with the onset of expression in haploid stages. Mol. Reprod. Dev. 65: 19–22, 2003. © 2003 Wiley‐Liss, Inc.
Development of the meiotic step in testes of pubertal rats: Comparison between the in vivo situation and under in vitro conditions
Authors: Perrard Marie‐HÉlÈne , Hue Dominique , Staub Christophe , Le Vern Yves , Kerboeuf Dominique , Durand Philippe
Published: March 2003

The present work aimed to compare some features of the meiotic process which develops in the testis of pubertal rats, in vivo and in vitro, paying special attention to the time‐course of the phenomenon. The differentiation of spermatocytes was assessed in testes of 20‐ to 46‐day‐old rats and in tubule segments of 20‐ or 28‐day‐old rats cultured over a 4‐week period. Very similar results were obtained in vivo and in vitro, during the first week of culture, when considering the changes in the cell populations of different ploidy, the gene expression of germ cells, the kinetics of differentiation of BrdU‐labeled early or middle pachytene spermatocytes and the levels of apoptosis in the different cell populations. However, during the second week of culture, the decrease in the proportion of the 4C cell population which was only slightly more marked than that observed in vivo between 27 and 34 days, was not associated with an increase in the 1C cell population as large as in vivo. This result could be explained partly by a high proportion of apoptotic 1C cells beyond one week of culture. Concomitantly, the rate of in vitro differentiation of BrdU‐labeled spermatocytes slowed down when reaching the stage of middle pachytene spermatocytes and BrdU‐labeled round spermatids were observed 6–11 days later than when BrdU‐labeled spermatocytes differentiated in vivo. Taken together, our results indicate that the bottleneck for the development of the meiotic cells in vitro is at the transition from middle to late pachytene spermatocytes. Hence, comparing the expression of locally produced regulatory molecules in vivo and in vitro at different days of culture should allow to identify key regulators of the meiotic step of spermatogenesis. Mol. Reprod. Dev. 65: 86–95, 2003. © 2003 Wiley‐Liss, Inc.
Family of Kunitz proteins from trophoblast: Expression of the trophoblast Kunitz domain proteins (TKDP) in cattle and sheep
Authors: MacLean James A. , Chakrabarty Anindita , Xie Sancai , Bixby James A. , Roberts R. Michael , Green Jonathan A.
Published: March 2003

Here we report the molecular cloning of several members of a family of novel proteins expressed by the ruminant trophoblast, known as the trophoblast Kunitz domain proteins (TKDPs). Each contains a carboxyl‐terminal module of ∼64 amino acids belonging to the Kunitz family of serine proteinase inhibitors. These Kunitz modules are preceded by one or more structurally related domains, each about 80 amino acids long. The function of these domains is unclear. The TKDPs differ considerably in sequence identity, with much of the diversity due to variability in the amino‐terminal domains. However, nine of the ten Kunitz domains described here are themselves unique, ranging in amino acid sequence identity from 90% to 53% to each other and averaging only about 50% identity with bovine pancreatic trypsin inhibitor (BPTI). The “warhead” P1 residues, which govern specificity, are themselves variable and include some unusual amino acids, such as Asn, Thr, and Ile, as well as the more common Lys. The Kunitz domains of three of the TKDPs lack the conserved cysteines at positions 14 and 38 (BPTI numbering) that normally contribute to the orientation of the inhibitory loop. Northern blotting and reverse transcription‐polymerase chain reaction (RT‐PCR) demonstrated that the TKDP genes do not exhibit identical expression patterns during trophoblast development, although mostly are expressed maximally during early pregnancy. It is possible that the TKDPs provide a broad range of specificities against maternal proteinases that might be damaging to the trophoblast during pregnancy. Mol. Reprod. Dev. 65: 30–40, 2003. © 2003 Wiley‐Liss, Inc.
Erasure of methylation imprinting of Igf2r during mouse primordial germ‐cell development
Authors: Sato Shun , Yoshimizu Tomomi , Sato Eimei , Matsui Yasuhisa
Published: March 2003

During germ cell differentiation in mice, the genome undergoes specific epigenetic modifications. These include demethylation of imprinted genes and subsequent establishment of parental allele‐specific methylation. The mouse Igf2r gene is an imprinted gene that shows maternal‐specific expression. Maternal‐specific methylation of differentially methylated region 2 (DMR2) of this gene may be necessary for its maternal‐specific expression. Before the allele‐specific methylation is established, DMR2 is demethylated in both male and female primordial germ cells (PGCs) by 13.5 days post coitum (dpc), indicating that the demethylation of this region occurs earlier in PGC development. The timing of the demethylation has been, however, unknown. In this study, we attempted to determine the timing of methylation erasure of Igf2r DMR2 in developing PGCs, using transgenic mice expressing green fluorescent protein specifically in the germ line. We purified migrating PGCs from the transgenic mice and examined the methylation status of DMR2. The results show that some CpG sites within DMR2 start demethylation at 9.5 dpc in some migrating PGCs, before the cells colonize genital ridges, and the progression of demethylation is rapid after colonization of the genital ridges. To examine whether the gonadal environment is involved in demethylation, we analyzed the methylation of DMR2 after culturing migrating PGCs in the absence of a gonadal environment. These culture experiments support the idea that a gonadal environment is not required for demethylation of the region in at least a fraction of PGCs. Mol. Reprod. Dev. 65: 41–50, 2003. © 2003 Wiley‐Liss, Inc.
Effect of roscovitine, a cdk1 inhibitor, and of the presence of oocyte on bovine cumulus cell expansion and cyclooxygenase‐2 expression
Authors: Vigneron C. , Nuttinck F. , Perreau C. , Reinaud P. , Charpigny G. , Mermillod P.
Published: March 2003

Oocyte maturation is accompanied by differentiation of surrounding cumulus cells. These cells produce hyaluronic acid (HA) and its storage in intercellular spaces results in expansion of the cells. The cumulus cells also accumulate cyclooxygenase‐2 (cox‐2) during maturation. Both expansion and cox‐2 storage are regulated by FSH and EGF. The aim of this study was to determine whether oocyte meiotic resumption is involved in the regulation of cumulus differentiation or not. We investigated the effects of roscovitine, a reversible inhibitor of meiosis resumption of cattle oocytes on EGF induced expansion and cox‐2 expression at the transcript and protein levels respectively (RT‐PCR and Western blot), in cumulus oocyte complexes (COCs) and cumulus complexes alone (CCs). EGF induced expansion and cox‐2 expression in both COCs and CCs. These effects were prevented by roscovitine, whether in the presence or in the absence of oocyte. However, the oocyte was essential for the reversibility of inhibition by roscovitine. In conclusion, our results indicate that i) oocyte secreted‐factors are not essential for cumulus expansion, and ii) roscovitine mediated inhibition of meiotic resumption also respects the functionality of the surrounding somatic cells. Mol. Reprod. Dev. 65: 114–121, 2003. © 2003 Wiley‐Liss, Inc.
Characterization of gene expression profiles in early bovine pregnancy using a custom cDNA microarray
Authors: Ishiwata Hiroko , Katsuma Susumu , Kizaki Keiichiro , Patel Osman V. , Nakano Hiroko , Takahashi Toru , Imai Kei , Hirasawa Akira , Shiojima Satoshi , Ikawa Hiroshi , Suzuki Yasuhito , Tsujimoto Gozoh , Izaike Yoshiaki , Todoroki Junichi , Hashizume Kazuyoshi
Published: March 2003

Gene expression analysis comparing nonpregnant with pregnant bovine uteri, including placenta, was performed with a custom cDNA microarray containing 1,933 independent genes. These genes were classified into six categories according to biological function, as follows: cell and tissue structural dynamics (108 genes), intercellular communication (221), intracellular metabolism (265), cell cycle and apoptosis (26), regulation of gene expression (113), expressed sequence tag (EST) and function unknown (617), and uncomplemented genes (583 clones). This array possessed bovine placental/endometrial specificity, as it included many pregnancy‐specific molecules, such as pregnancy‐associated glycoprotein‐1 (PAGs), placental lactogen (PLs), and prolactin‐related protein‐1 (PRPs). A total of 77 genes were induced and 12 repressed in the placenta/endometrium. Our results point to a fundamental role for bovine placental‐specific genes such as PAGs, PLs, and PRPs, in implantation and placentogenesis, and document that cDNA microarray analysis from bovine placenta/endometrium is possible and is a specific tool for monitoring genome‐wide gene expression during the establishment and maintenance of pregnancy. Mol. Reprod. Dev. 65: 9–18, 2003. © 2003 Wiley‐Liss, Inc.
Nucleolar proteins and ultrastructure in bovine in vivo developed, in vitro produced, and parthenogenetic cleavage‐stage embryos
Authors: Laurincik J. , Schmoll F. , Mahabir E. , Schneider H. , Stojkovic M. , Zakhartchenko V. , Prelle K. , Hendrixen P.J.M. , Voss P.L.A.M. , Moeszlacher G.G. , Avery B. , Dieleman S.J. , Besenfelder U. , Müller M. , Ochs R.L. , Wolf E. , Schellander K. , Maddox‐Hyttel P.
Published: March 2003

In the present study, ribosomal RNA (rRNA) gene activation, monitored through nucleolus development, was studied by autoradiography following 3H‐uridine incubation, transmission electron microscopy, and immunofluorescence confocal laser scanning microscopy of key nucleolar proteins involved in rRNA transcription (topoisomerase I, upstream binding factor, and RNA polymerase I) and processing (fibrillarin, nucleolin, and nucleophosmin) in in vivo developed, in vitro produced, and parthenogenetic bovine embryos. In general, in vivo developed embryos displayed formation of fibrillo‐granular nucleoli during the 4th post‐fertilization cell cycle. During the previous stages of development, nucleolus precursor bodies (NPBs) were observed. However, on some occasions the initial steps of nucleolus formation were observed already at the 2‐ and 4‐cell stage in cases where such embryos were collected from superovulated animals together with later embryonic stages presenting nucleolar development and autoradiographic labeling. The in vitro produced embryos displayed very synchronous formation of fibrillo‐granular nucleoli and autoradiographic labeling during the 4th cell cycle. In vivo developed and in vitro produced embryos displayed allocation of nucleolar proteins to fibrillar and granular compartments of the developing nucleoli during the 4th cell cycle. The parthenogenetic embryos typically displayed formation of fibrillo‐ granular nucleoli during the 5th cell cycle and autoradiographic labeling was not observed until the morula stage. Moreover, the 1‐, 2‐, and 4‐cell parthenogenetic embryos practically lacked NPBs. On the other hand, parthenogenetic embryos displayed allocation of nucleoar proteins to nuclear entities during the 4th cell cycle. In conclusion, both in vivo developed and in vitro produced bovine embryos displayed activation of transcription and nucleolar development during the 4th cell cycle. However, in vivo developed embryos flushed together with later developmental stages displayed premature activation of these processes. Parthenogenetic bovine embryos, on the other hand, displayed a delayed activation. Mol. Reprod. Dev. 65: 73–85, 2003. © 2003 Wiley‐Liss, Inc.
Immunolocalization of multiple Gα subunits in mammalian spermatozoa and additional evidence for Gαs
Authors: Baxendale Rhona W. , Fraser Lynn R.
Published: March 2003

Like somatic cells, mammalian spermatozoa appear to contain several different heterotrimeric G protein α‐subunits that could mediate specialized cell responses. However, the precise Gα subunits present, their subcellular location and their possible roles are still incompletely defined. In this study, using commercially available specific antibodies, we have shown by immunoblotting that Gαs is present in human and mouse sperm lysates. Immunolocalization using intact spermatozoa from both species revealed this protein to be in the acrosomal cap region and the flagellum, particularly the principal piece. Treatment of permeabilized mouse spermatozoa with cholera toxin led to enhanced ADP‐ribosylation of a protein the same size as Gαs, as well as an increase in cAMP, providing further proof for Gαs. Evidence for the presence and distinct localizations of Gαi2, Gαi3, Gαo, Gαq/11, and Gαolf was also obtained. Of particular interest was Gαi2 which, like Gαs, was present in the acrosomal cap region and flagellum, the same regions where stimulatory and inhibitory adenosine receptors are localized. These observations are consistent with our hypothesis that G proteins mediate adenosine receptor modulation of adenylyl cyclase, with consequent alterations in cAMP production, apparently crucial for the spermatozoon's acquisition and maintenance of fertilizing ability. Mol. Reprod. Dev. 65: 104–113, 2003. © 2003 Wiley‐Liss, Inc.
Nuclear envelope proteins and neuromuscular diseases
Authors: Östlund Cecilia , Worman Howard J.
Published: March 2003

Several neuromuscular diseases are caused by mutations in emerin and A‐type lamins, proteins of the nuclear envelope. Emery–Dreifuss muscular dystrophy is caused by mutations in emerin (X‐linked) or A‐type lamins (autosomal dominant). Mutations in A‐type lamins also cause limb‐girdle muscular dystrophy type 1B, dilated cardiomyopathy with conduction defect, and Charcot–Marie–Tooth disorder type 2B1. They also cause partial lipodystrophy syndromes. The functions of emerin and A‐type lamins and the mechanisms of how mutations in these proteins cause tissue‐specific diseases are not well understood. The mutated proteins may cause structural damage to cells but may also affect processes such as gene regulation. This review gives an overview of this topic and describes recent advances in identification of disease‐causing mutations, studies of cells and tissues from subjects with these diseases, and animal and cell culture models. Muscle Nerve 27: 393–406, 2003
CBA/J mice infected with Trypanosoma cruzi: An experimental model for inflammatory myopathies
Authors: Andersson John , Englund Pernilla , Sunnemark Dan , Dahlstedt Anders , Westerblad Håkan , Nennesmo Inger , Örn Anders , Lundberg Ingrid E.
Published: March 2003

Idiopathic inflammatory myopathies are inflammatory disorders of unknown origin, characterized by muscle weakness. The aim of our study was to establish and characterize an animal model for chronic inflammatory myopathy which would permit investigations of the role of T‐cells in chronic myositis as well as of the mechanisms for muscle weakness in chronic inflammatory muscle disorders. CBA/J mice were infected with the protozoan parasite Trypanosoma cruzi. Immunohistochemistry was used to characterize the distribution and composition of inflammatory infiltrates, and demonstrated a chronic focal inflammation comprised mainly of T‐cells and macrophages in infected mice. The inflammatory infiltrates were predominantly found in the endomysium and, to a lesser extent, in perivascular areas. CD8+ T‐cells were found to have invaded nonnecrotic muscle fibers. Degenerating muscle fibers were also found, as well as an increased number of central muscle nuclei. The murine model described in this article may be useful in studying certain aspects of idiopathic inflammatory myopathies such as the role of T‐cells in chronic muscle inflammation and chronic myocytotoxicity. Muscle Nerve 27:442–448, 2003
Cross‐bridge mechanisms of muscle weakness in multiple sclerosis
Authors: Garner Dena J.P. , Widrick Jeffrey J.
Published: March 2003

Vastus lateralis muscle biopsies were obtained from six individuals with multiple sclerosis (MS) having an Expanded Disability Status Score of 4.75 ± 0.28, and from six age‐ and gender‐matched individuals without MS. Biopsies from the MS group showed fewer fibers (31 ± 4 vs. 46 ± 4%) containing the type IIa myosin heavy chain (MHC) isoform exclusively. However, the percentage of fibers coexpressing type IIa and IIx MHC increased in direct proportion with MS disability status. The average unloaded shortening velocity of skinned fibers containing type I or IIa MHC did not differ between subject groups. Peak Ca2+‐activated force was 11–13% lower in fibers from the MS group due to atrophy (type I and IIa fibers) and reduced specific force (type I fibers). Increasing intracellular inorganic phosphate (0–30 mM) or hydrogen ion (pH 7.0–6.2) reduced Ca2+‐activated force in a manner that was independent of MS status. Thus, fibers from the MS group showed a subtle shift in fast MHC isoform coexpression and a modest reduction in cross‐bridge number, density, or average force, with no change in maximal cross‐bridge cycling rate or susceptibility to intracellular metabolites. These changes explain part of the muscle weakness and fatigue experienced by individuals with MS. Muscle Nerve 27: 456–464, 2003
Diagnostic value of nerve biopsy for atypical chronic inflammatory demyelinating polyneuropathy: Evaluation of eight cases
Authors: Vallat Jean‐Michel , Tabaraud François , Magy Laurent , Torny Frédéric , Bernet‐Bernady Patricia , Macian Francisco , Couratier Philippe
Published: March 2003

The diagnosis of chronic inflammatory demyelinating polyneuropathy (CIDP) relies primarily on clinical and electrophysiologic examination, but the nerve biopsy findings may be supportive, especially in atypical cases. In order to define the usefulness of nerve biopsy in this disease, we retrospectively studied 44 consecutive patients whom we classified as having CIDP on pathological grounds. We found that 8 of these 44 patients had pathological findings indicative of CIDP but did not meet any of the usually accepted electrophysiological criteria for its diagnosis. Among these eight patients, five responded favorably to conventional therapy. All of these eight patients had an electrophysiological pattern of generalized axonopathy with additional subtle findings suggestive of demyelination that prompted us to perform a nerve biopsy. Our data suggest that a significant number of patients with unrecognized CIDP are erroneously classified as having chronic idiopathic axonal polyneuropathy. CIDP should be suspected if the electrophysiological examination displays subtle abnormalities suggestive of demyelination, even in the presence of a prominent axonal pattern. Nerve biopsy in these patients may reveal abnormalities suggestive of CIDP and guide therapeutic options. Muscle Nerve 27: 478–485, 2003
Abnormal coupling of knee and hip moments during maximal exertions in persons with cerebral palsy
Authors: Thelen Darryl D. , Riewald Scott A. , Asakawa Deanna S. , Sanger Terence D. , Delp Scott L.
Published: March 2003

The motions of lower‐limb extension, adduction, and internal rotation are frequently coupled in persons with cerebral palsy (CP) and are commonly referred to as an extension synergy. However, the underlying joint moments that give rise to these coupled motions are not well understood. We hypothesized that maximal voluntary exertions in a direction of one component of a synergy (e.g., hip extension) would result in the concurrent presence of other components of the synergy in subjects with CP but not in control subjects. To test this hypothesis, we measured three‐dimensional moments about the hip and knee as nine subjects with spastic CP and six control subjects performed maximal isometric exertions of the hip and knee flexors and extensors. During maximal hip extension exertions, control subjects simultaneously generated a knee flexion moment, whereas CP subjects generated a knee extension moment (P < 0.05) and a larger hip internal rotation moment than did controls (P < 0.05). During maximal knee extension exertions, control subjects generated a hip flexion moment, whereas CP subjects generated a hip extension moment (P < 0.05). The patterns of joint moments generated by CP subjects are consistent with an extension synergy and may underlie the coupled motion patterns of the lower extremity in such persons. Muscle Nerve 27: 486–493, 2003