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Background: During the preschool years, children develop abilities and skills in areas crucial for later success in life. These abilities include language, executive functions, attention, and socioemotional skills. The pedagogical methods used in preschools hold the potential to enhance these abilities, but our knowledge of which pedagogical practices aid which abilities, and for which children, is limited. The aim of this paper is to describe an intervention study designed to evaluate and compare two pedagogical methodologies in terms of their effect on the above-mentioned skills in Swedish preschool children., Method: The study is a randomized control trial (RCT) where two pedagogical methodologies were tested to evaluate how they enhanced children's language, executive functions and attention, socioemotional skills, and early maths skills during an intensive 6-week intervention. Eighteen preschools including 28 units and 432 children were enrolled in a municipality close to Stockholm, Sweden. The children were between 4;0 and 6;0 years old and each preschool unit was randomly assigned to either of the interventions or to the control group. Background information on all children was collected via questionnaires completed by parents and preschools. Pre- and post-intervention testing consisted of a test battery including tests on language, executive functions, selective auditive attention, socioemotional skills and early maths skills. The interventions consisted of 6 weeks of intensive practice of either a socioemotional and material learning paradigm (SEMLA), for which group-based activities and interactional structures were the main focus, or an individual, digitally implemented attention and math training paradigm, which also included a set of self-regulation practices (DIL). All preschools were evaluated with the ECERS-3., Discussion: If this intervention study shows evidence of a difference between group-based learning paradigms and individual training of specific skills in terms of enhancing children's abilities in fundamental areas like language, executive functions and attention, socioemotional skills and early math, this will have big impact on the preschool agenda in the future. The potential for different pedagogical methodologies to have different impacts on children of different ages and with different backgrounds invites a wider discussion within the field of how to develop a preschool curriculum suited for all children.

Mammals evolved from oviparous reptiles that laid eggs in a dry, terrestrial environment, thus requiring large amounts of yolk to support development and tough, outer coats to protect them. Eutherian mammals such as humans and mice exhibit an "extreme" form of viviparity in which yolk and conceptus coats have become largely redundant. However, the "other" mammals-monotremes and marsupials-have retained and modified some features of reptilian development that provide valuable insights into the evolution of viviparity in mammals. Most striking of these are the conceptus coats, which include the zona pellucida, the mucoid coat, and the shell coat. We discuss current knowledge of these coats in monotremes and marsupials, their possible roles, and recently identified components such as the zona pellucida protein ZPAX, conceptus coat mucin (CCM), and nephronectin (NPNT)., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Marsupials and monotremes differ from eutherian mammals in many features of their reproduction and development. Some features appear to be representative of transitional stages in evolution from therapsid reptiles to humans and mice, particularly with respect to the extraembryonic tissues that have undergone remarkable modifications to accommodate reduced egg size and quantity of yolk/deutoplasm, and increasing emphasis on viviparity and placentation. Trophoblast and hypoblast contribute the epithelial layers in most of the extraembryonic membranes and are the first two lineages to differentiate from the embryonic lineage. How they are specified varies greatly among mammals, perhaps largely due to heterochrony in the stage at which they must function. Differences probably also exist in the stage at which lineages are specified relative to the stage at which they fully commit to differentiation. The dogma of sequential commitment to trophoblast and hypoblast with progressive loss of potency may not be a fundamental feature of early mammalian development, but merely a recently acquired developmental pattern in eutherians, or at least mice., (© 2018 Elsevier Inc. All rights reserved.)

The ambush bugs (Heteroptera: Reduviidae: Phymatinae) are a diverse clade of predators known for their cryptic hunting behavior and morphologically diverse raptorial forelegs. Despite their striking appearance, role as pollinator predators, and intriguing biogeographic distribution, phylogenetic relationships within Phymatinae are largely unknown and the evolutionary history of the subfamily has remained in the dark. We here utilize the most extensive molecular phylogeny of ambush bugs to date, generated from a 3328 base pair molecular dataset, to refine our understanding of phymatine relationships, estimate dates of divergence (BEAST 2), and uncover historical biogeographic patterns (S-DIVA and DEC). This taxon set (39 species of Phymatinae and six outgroups) allowed reevaluation of the proposed sister group of Phymatinae and tribal-level relationships within the group, and for the first time proposes species-level relationships within Phymata Latreille, the largest genus of ambush bugs (∼109spp.). Available evidence suggests that Phymata originated in the Neotropical region, with subsequent dispersals to the Nearctic and Palearctic regions. This study provides a framework for future research investigating the evolutionary history of ambush bugs, as well as ecological and microevolutionary investigations., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The control of reactivation from embryonic diapause in the tammar wallaby (Macropus eugenii) involves sequential activation of the corpus luteum, secretion of progesterone that stimulates endometrial secretion and subsequent changes in the uterine environment that activate the embryo. However, the precise signals between the endometrium and the blastocyst are currently unknown. In eutherians, both the phospholipid Paf and its receptor, platelet-activating factor receptor (PTAFR), are present in the embryo and the endometrium. In the tammar, endometrial Paf release in vitro increases around the time of the early progesterone pulse that occurs around the time of reactivation, but whether Paf can reactivate the blastocyst is unknown. We cloned and characterised the expression of PTAFR in the tammar embryo and endometrium at entry into embryonic diapause, during its maintenance and after reactivation. Tammar PTAFR sequence and protein were highly conserved with mammalian orthologues. In the endometrium, PTAFR was expressed at a constant level in the glandular epithelium across all stages and in the luminal epithelium during both diapause and reactivation. Thus, the presence of the receptor appears not to be a limiting factor for Paf actions in the endometrium. However, the low levels of PTAFR in the embryo during diapause, together with its up-regulation and subsequent internalisation at reactivation, supports earlier results suggesting that endometrial Paf could be involved in reactivation of the tammar blastocyst from embryonic diapause.

Background: Pluripotency is a fundamental property of early mammalian development but it is currently unclear to what extent its cellular mechanisms are conserved in vertebrates or metazoans. POU5F1 and POU2 are the two principle members constituting the class V POU domain family of transcription factors, thought to have a conserved role in the regulation of pluripotency in vertebrates as well as germ cell maintenance and neural patterning. They have undergone a complex pattern of evolution which is poorly understood and controversial., Results: By analyzing the sequences of POU5F1, POU2 and their flanking genes, we provide strong indirect evidence that POU5F1 originated at least as early as a common ancestor of gnathostomes but became extinct in a common ancestor of teleost fishes, while both POU5F1 and POU2 survived in the sarcopterygian lineage leading to tetrapods. Less divergent forms of POU5F1 and POU2 appear to have persisted among cartilaginous fishes., Conclusions: Our study resolves the controversial evolutionary relationship between teleost pou2 and tetrapod POU2 and POU5F1, and shows that class V POU transcription factors have existed at least since the common ancestor of gnathostome vertebrates. It provides a framework for elucidating the basis for the lineage-specific extinctions of POU2 and POU5F1.

The preimplantation period of mouse early embryonic development is devoted to the specification of two extraembryonic tissues and their spatial segregation from the pluripotent epiblast. During this period two cell fate decisions are made while cells gradually lose their totipotency. The first fate decision involves the segregation of the extraembryonic trophectoderm (TE) lineage from the inner cell mass (ICM); the second occurs within the ICM and involves the segregation of the extraembryonic primitive endoderm (PrE) lineage from the pluripotent epiblast (EPI) lineage, which eventually gives rise to the embryo proper. Multiple determinants, such as differential cellular properties, signaling cues and the activity of transcriptional regulators, influence lineage choice in the early embryo. Here, we provide an overview of our current understanding of the mechanisms governing these cell fate decisions ensuring proper lineage allocation and segregation, while at the same time providing the embryo with an inherent flexibility to adjust when perturbed., (Copyright © 2013 Wiley Periodicals, Inc.)

Early cell lineage specification in eutherian mammals results in the formation of a pluripotent inner cell mass (ICM) and trophoblast. By contrast, marsupials have no ICM. Here, we present the first molecular analysis of mechanisms of early cell lineage specification in a marsupial, the tammar wallaby. There was no overt differential localisation of key lineage-specific transcription factors in cleavage and early unilaminar blastocyst stages. Pluriblast cells (equivalent to the ICM) became distinguishable from trophoblast cells by differential expression of POU5F1 and, to a greater extent, POU2, a paralogue of POU5F1. Unlike in the mouse, pluriblast-trophoblast differentiation coincided with a global nuclear-to-cytoplasmic transition of CDX2 localisation. Also unlike in the mouse, Hippo pathway factors YAP and WWTR1 showed mutually distinct localisation patterns that suggest non-redundant roles. NANOG and GATA6 were conserved as markers of epiblast and hypoblast, respectively, but some differences to the mouse were found in their mode of differentiation. Our results suggest that there is considerable evolutionary plasticity in the mechanisms regulating early lineage specification in mammals.

Cell differentiation during pre-implantation mammalian development involves the formation of two extra-embryonic lineages: trophoblast and primitive endoderm (PrE). A subset of cells within the inner cell mass (ICM) of the blastocyst does not respond to differentiation signals and forms the pluripotent epiblast, which gives rise to all of the tissues in the adult body. How this group of cells is set aside remains unknown. Recent studies documented distinct sequential phases of marker expression during the segregation of epiblast and PrE within the ICM. However, the connection between marker expression and lineage commitment remains unclear. Using a fluorescent reporter for PrE, we investigated the plasticity of epiblast and PrE precursors. Our observations reveal that loss of plasticity does not coincide directly with lineage restriction of epiblast and PrE markers, but rather with exclusion of the pluripotency marker Oct4 from the PrE. We note that individual ICM cells can contribute to all three lineages of the blastocyst until peri-implantation. However, epiblast precursors exhibit less plasticity than precursors of PrE, probably owing to differences in responsiveness to extracellular signalling. We therefore propose that the early embryo environment restricts the fate choice of epiblast but not PrE precursors, thus ensuring the formation and preservation of the pluripotent foetal lineage.

During preimplantation mouse development, the inner cell mass (ICM) differentiates into two cell lineages--the epiblast and the primitive endoderm (PrE)--whose precursors are identifiable by reciprocal expression of Nanog and Gata6, respectively. PrE formation depends on Nanog by a non-cell-autonomous mechanism. To decipher early cell- and non-cell-autonomous effects, we performed a mosaic knockdown of Nanog and found that this is sufficient to induce a PrE fate cell autonomously. Strikingly, in Nanog null embryos, Gata6 expression is maintained, showing that initiation of the PrE program is Nanog independent. Treatment of Nanog null embryos with pharmacological inhibitors revealed that RTK dependency of Gata6 expression is initially direct but later indirect via Nanog repression. Moreover, we found that subsequent expression of Sox17 and Gata4--later markers of the PrE--depends on the presence of Fgf4 produced by Nanog-expressing cells. Thus, our results reveal three distinct phases in the PrE differentiation program., (Copyright © 2011 Elsevier Inc. All rights reserved.)

DDX4 (VASA) is an RNA helicase expressed in the germ cells of all animals. To gain greater insight into the role of this gene in mammalian germ cell development, we characterized DDX4 in both a marsupial (the tammar wallaby) and a monotreme (the platypus). DDX4 is highly conserved between eutherian, marsupial, and monotreme mammals. DDX4 protein is absent from tammar fetal germ cells but is present from Day 1 postpartum in both sexes. The distribution of DDX4 protein during oogenesis and spermatogenesis in the tammar is similar to eutherians. Female tammar germ cells contain DDX4 protein throughout all stages of postnatal oogenesis. In males, DDX4 is in gonocytes, and during spermatogenesis it is present in spermatocytes and round spermatids. A similar distribution of DDX4 occurs in the platypus during spermatogenesis. There are several DDX4 isoforms in the tammar, resulting from both pre- and posttranslational modifications. DDX4 in marsupials and monotremes has multiple splice variants and polyadenylation motifs. Using in silico analyses of genomic databases, we found that these previously unreported splice variants also occur in eutherians. In addition, several elements implicated in the control of Ddx4 expression in the mouse, including RGG (arginine-glycine-glycine) and dimethylation of arginine motifs and CpG islands within the Ddx4 promoter, are also highly conserved. Collectively these data suggest that DDX4 is essential for the regulation of germ cell proliferation and differentiation across all three extant mammalian groups-eutherians, marsupials, and monotremes.

Successful maintenance, survival and maturation of gametes rely on bidirectional communication between the gamete and its supporting cells. Before puberty, factors from the gamete and its supporting cells are necessary for spermatogonial stem cell and primordial follicle oocyte maintenance. Following gametogenesis, gametes rely on factors and nutrients secreted by cells of the reproductive tracts, the epididymis and/or oviduct, to complete maturation. Despite extensive studies on female and male reproduction, many of the molecular mechanisms of germ cell maintenance remain relatively unknown, particularly in marsupial species. We present the first study and characterisation of a novel primary miRNA transcript, pri-miR-16c, in the marsupial, the stripe-faced dunnart. Bioinformatic analysis showed that its predicted processed miRNA - miR-16c - is present in a wide range of vertebrates, but not eutherians. In situ hybridisation revealed dunnart pri-miR-16c expression in day 4 (primordial germ cells) and day 7 (oogonia) pouch young, in primary oocytes and follicle cells of primordial follicles but then only in follicle cells of primary, secondary and antral follicles in adult ovaries. In the adult testis, pri-miR-16c transcripts were present in the cytoplasm of spermatogonial cells. The oviduct and the epididymis both showed expression, but not any other somatic tissues examined or conceptuses during early embryonic development. This pattern of expression suggests that pri-miR-16c function may be associated with gamete maintenance, possibly through mechanisms involving RNA transfer, until the zygote enters the uterus at the pronuclear stage.

Background: The vomeronasal organ (VNO) detects pheromones via two large families of vomeronasal receptors: vomeronasal receptor 1 (V1R) and vomeronasal receptor 2 (V2R). Both VRs have a common receptor activation cascade involving transient receptor potential channel, subfamily C, member 2 (TRPC2)., Results: We characterised the TRPC2 locus in a marsupial, the tammar wallaby (Macropus eugenii), and identified two independently regulated genes not previously recognised as distinct. 3'-located exons comprise bona fide TRPC2 whilst 5'-located exons, previously identified as part of TRPC2, comprise a distinct gene, which we term XNDR (XRCC1 N-terminal domain-related). The two genes show contrasting expression patterns in the tammar: TRPC2 is specifically expressed in adult and developing VNO, whereas XNDR is widely expressed in many tissues suggesting a non-VNO-specific role. Strong expression of TRPC2 was detected only after about day 30 post-partum, suggesting that the VNO may not be functional during early pouch life of the tammar. Similarly restricted expression of TRPC2 and widespread expression of XNDR was also detected in the platypus. Bioinformatic analysis of the genomes of a wide range of species suggests that the identity of XNDR and TRPC2 as distinct genes is conserved among vertebrates. Finally, we analysed the promoter of mammalian TRPC2 and identified a conserved binding site for NHLH1, a transcription factor previously implicated in VNO receptor neuron development., Conclusions: Two functionally distinct vertebrate genes-XNDR and TRPC2 - occupy a genomic locus that was previously defined as a single gene in the mouse. The former is widely expressed with a putative role in DNA repair, while the latter shows VNO-specific expression under the probable regulation of NHLH1.

Background: ATRX is a tightly-regulated multifunctional protein with crucial roles in mammalian development. Mutations in the ATRX gene cause ATR-X syndrome, an X-linked recessive developmental disorder resulting in severe mental retardation and mild alpha-thalassemia with facial, skeletal and genital abnormalities. Although ubiquitously expressed the clinical features of the syndrome indicate that ATRX is not likely to be a global regulator of gene expression but involved in regulating specific target genes. The regulation of ATRX expression is not well understood and this is reflected by the current lack of identified upstream regulators. The availability of genomic data from a range of species and the very highly conserved 5' regulatory regions of the ATRX gene has allowed us to investigate putative transcription factor binding sites (TFBSs) in evolutionarily conserved regions of the mammalian ATRX promoter., Results: We identified 12 highly conserved TFBSs of key gene regulators involved in biologically relevant processes such as neural and testis development and alpha-globin regulation., Conclusions: Our results reveal potentially important regulatory elements in the ATRX gene which may lead to the identification of upstream regulators of ATRX and aid in the understanding of the molecular mechanisms that underlie ATR-X syndrome.

POU5F1 (OCT4) encodes a master regulator of pluripotency that is present in all mammals. A paralogue, POU2, is also present in the genomes of marsupials and monotremes and is an orthologue of zebrafish pou2 and chicken POUV. We explored the evolution of class V POU domain transcription factors and show that POU5F1 arose by gene duplication of pou2 early in the evolution of tetrapods and is not mammal-specific, as previously thought. Instead, either POU5F1 or POU2/POUV has become extinct independently in various lineages, although all gnathostomes appear to possess at least one or the other. In the tammar wallaby, POU5F1 expression is limited to pluripotent cell types (embryonic tissues and germ cells). POU2 is similarly expressed in pluripotent tissues but is also expressed in a broad range of adult tissues. Thus, unlike POU5F1, the role of POU2 may not be restricted to pluripotent cell types but could have a related function such as maintaining multipotency in adult stem cells.

The first two lineages to differentiate from a pluripotent cell population during mammalian development are the extraembryonic trophectoderm (TE) and the primitive endoderm (PrE). Whereas the mechanisms of TE specification have been extensively studied, segregation of PrE and the pluripotent epiblast (EPI) has received comparatively little attention. A current model of PrE specification suggests PrE precursors exhibit an apparently random distribution within the inner cell mass of the early blastocyst and then segregate to their final position lining the cavity by the late blastocyst. We have identified platelet-derived growth factor receptor alpha (Pdgfralpha) as an early-expressed protein that is also a marker of the later PrE lineage. By combining live imaging of embryos expressing a histone H2B-GFP fusion protein reporter under the control of Pdgfra regulatory elements with the analysis of lineage-specific markers, we investigated the events leading to PrE and EPI lineage segregation in the mouse, and correlated our findings using an embryo staging system based on total cell number. Before blastocyst formation, lineage-specific factors are expressed in an overlapping manner. Subsequently, a gradual progression towards a mutually exclusive expression of PrE- and EPI-specific markers occurs. Finally, cell sorting is achieved by a variety of cell behaviours and by selective apoptosis.

Background: To date, the earliest stage at which the orientation of the anterior-posterior axis in the mouse embryo is distinguishable by asymmetric gene expression is shortly after E5.5. At E5.5, prospective anterior markers are expressed at the distal tip of the embryo, whereas prospective posterior markers are expressed more proximally, close to the boundary with the extraembryonic region., Results: To contribute to elucidating the mechanisms underlying the events involved in early patterning of the mouse embryo, we have carried out a microarray screen to identify novel genes that are differentially expressed between the distal and proximal parts of the E5.5 embryo. Secondary screening of resulting candidates by in situ hybridisation at E5.5 and E6.5 revealed novel expression patterns for known and previously uncharacterised genes, including Peg10, Ctsz1, Cubilin, Jarid1b, Ndrg1, Sfmbt2, Gjb5, Talia and Plet1. The previously undescribed gene Talia and recently identified Plet1 are expressed specifically in the distal-most part of the extraembryonic ectoderm, adjacent to the epiblast, and are therefore potential candidates for regulating early patterning events. Talia and the previously described gene XE7 define a gene family highly conserved among metazoans and with a predicted protein structure suggestive of a post-transcriptional regulative function, whilst Plet1 appears to be mammal-specific and of unknown function., Conclusion: Our approach has allowed us to compare expression between dissected parts of the egg cylinder and has identified multiple genes with novel expression patterns at this developmental stage. These genes are potential candidates for regulating tissue interactions following implantation.

Generation of inside cells that develop into inner cell mass (ICM) and outside cells that develop into trophectoderm is central to the development of the early mouse embryo. Critical to this decision is the development of cell polarity and the associated asymmetric (differentiative) divisions of the 8-cell-stage blastomeres. The underlying molecular mechanisms for these events are not understood. As the Par3/aPKC complex has a role in establishing cellular polarity and division orientation in other systems, we explored its potential function in the developing mouse embryo. We show that both Par3 and aPKC adopt a polarized localization from the 8-cell stage onwards and that manipulating their function re-directs cell positioning and consequently influences cell fate. Injection of dsRNA against Par3 or mRNA for a dominant negative form of aPKC into a random blastomere at the 4-cell stage directs progeny of the injected cell into the inside part of the embryo. This appears to result from both an increased frequency by which such cells undertake differentiative divisions and their decreased probability of retaining outside positions. Thus, the natural spatial allocation of blastomere progeny can be over-ridden by downregulation of Par3 or aPKC, leading to a deceased tendency for them to remain outside and so develop into trophectoderm. In addition, this experimental approach illustrates a powerful means of manipulating gene expression in a specific clonal population of cells in the preimplantation embryo.

Objective: To compare treadmill training with partial body-weight support (TT-BWS) and conventional physical therapy (PT) in ambulatory patients with hip arthroplasty., Design: Randomized controlled trial., Setting: Rehabilitation center., Participants: Eighty patients with a fully loadable implant who could walk independently with crutches after unilateral total hip arthroplasty were randomized to receive either TT-BWS (treatment group) or conventional PT (controls), for 10 working days., Interventions: Each patient received 45 minutes of individualized PT, either treadmill training plus PT in the experimental or PT alone in the control group., Main Outcome Measures: The Harris score, recorded by blind assessors, served as the primary outcome measure. Secondary outcome measures were the hip extension deficit, gait velocity, gait symmetry, affected hip abductor power; hip abductor amplitude of electromyographic activation; and the interval from surgery to abandoning crutches., Results: At the end of training, the treatment group's Harris score was 13.6 points higher (P<.0001) than the control group's score. Further, hip extension deficit was 6.8 degrees less (P<.0001), gait symmetry was 10% greater (P=.001), affected hip abductor was stronger (Medical Research Council grades 4.24 vs 3.73; P<.0001), and the amplitude of gluteus medius activity was 41.5% greater (P=.001) than those measures for controls. Gait velocity did not differ in the 2 groups. These significant differences in favor of the treatment group persisted at 3 and 12 months. The treatment group abandoned crutches sooner than the control group (3 vs 8wk). In the treatment group, 39 patients finished treatment, 35 appeared at 3, and 26 at 12 months for follow-up. In the control group, the corresponding numbers were 40, 35, and 24 patients, respectively., Conclusion: TT-BWS is more effective than conventional PT at restoring symmetrical independent walking after hip replacement.

Treadmill therapy with partial BWS is a promising new approach to improve gait ability after stroke. This task-specific approach enables nonambulatory patients the repetitive practice of complex gait cycles instead of single-limb gait-preparatory maneuvers. Patients walk more symmetrically with less spasticity and better cardiovascular efficiency on the treadmill than with floor walking. Several controlled, clinical studies have shown the potential of treadmill training as a therapeutic intervention for nonambulatory patients with chronic stroke-related hemiplegia. Furthermore, controlled trials in acute stroke survivors have shown that treadmill training is as effective as other physiotherapy approaches that stress the repetitive practice of gait. Controlled multicenter trials comparing locomotor training with conventional therapy will be forthcoming. An electromechanical gait trainer that relieves the strenuous effort of the therapists and provides control of the trunk in a phase-dependent manner is a new technical alternative for gait training in severely impaired stroke patients.

Background: Because nutritional intolerances generally are difficult to be diagnosed, Applied Kinesiology (AK) often is discussed as an alternative in this field., Aim: The uncertain scientific situation makes it necessary to assess the diagnostic value of AK. The method was compared with established laboratory tests such as RAST, lactose breath hydrogen test and a specific IgG test (Cytolisa)., Methods: 315 children and adolescents, 0-17 years old, with different chronic diseases such as headache, abdominal pain, chronic eczema, hyperactivity, and bronchial asthma were studied by means of AK for 2 years., Results: We found a moderate test-retest reliability (intraclass-kappa 0.62) but no inter-tester reliability (intraclass-kappa -0.01). Moreover, there was no statistically significant agreement with either RAST and Cytolisa (sensitivity 73.6%, specificity 45.2%) or lactose breath hydrogen test (sensitivity 77.1%, specificity 43.2%)., Conclusions: In general, AK cannot be recommended for diagnosing nutritional intolerance. However, due to its high sensitivity, it could be a valuable tool to give some preliminary results., (Copyright 2001 S. Karger GmbH, Freiburg)

Cytochrome P450 enzymes metabolize various endogenous and exogenous small molecular weight compounds. Transport-associated proteins, such as P-glycoprotein, multidrug resistance-associated protein and lung resistance protein are overexpressed in drug-resistant cell lines, as well as in human tumors from various histologic origins, including malignant melanoma. Little is known about the expression and function of cytochrome enzymes and multidrug resistance-associated transport proteins in human skin; therefore, the aim of this study was to analyze the expression pattern of cytochrome enzymes and multidrug resistance-associated transport proteins in proliferating human epidermal keratinocytes under constitutive conditions and after induction with various inducers. Reverse transcription-polymerase chain reaction revealed constitutive expression of cytochromes 1A1, 1B1, 2B6, 2E1, and 3A5 in keratinocytes and showed expression of cytochrome 3A4 after incubation with dexamethasone. The expression of cytochrome 1A1 was enhanced on the mRNA level after induction with benzanthracene. Reverse transcription-polymerase chain reaction analysis of the multidrug resistance-associated transport proteins revealed constitutive expression of multidrug resistance-associated proteins 1 and 3-6, and lung resistance protein in human epithelial keratinocytes and was negative for multidrug resistance 1 and 2. Expression of 1 was seen after induction with dexamethasone. Reverse transcription-polymerase chain reaction results were confirmed by immunoblots which showed expression of cytochromes 1A1, 2B6, 2E1, and 3A, multidrug resistance-associated proteins 1, 3, and 5 as well as multidrug resistance 1 after induction with dexamethasone. Immunohistology showed positive immunofluorescence in skin specimens for cytochromes 1A1, 2B6, 2E1, and 3A and multidrug resistance-associated protein 1 and multidrug resistance 1. Constitutive activity of cytochrome 1A1, 2B, 2E1, and 3A enzymes was measured by catalytic assays. These results show that keratinocytes of the human skin express various transport-associated enzymes and detoxifying metabolic enzymes. Previous studies have revealed that cytochrome enzymes and transport-associated proteins play complementary parts in drug disposition by biotransformation (phase I) and anti-transport (phase III) and act synergistically as a drug bioavailability barrier.

Marsupial ova are slightly larger on average than those of eutherians. They have often been described as "yolky", although the nature and roles of their contents in later development are poorly understood. In the ova of all marsupial species studied, large quantities of electron-lucent vesicles are present. This study describes the ultrastructure of oogenesis in the brushtail possum, Trichosurus vulpecula. Oocytes of primordial follicles contain a variety of organelles of unknown function, some of which have been described previously in bandicoots. Large electron-lucent vesicles begin to accumulate after the transition from primordial follicle to primary follicle and are derived at least partly from the Golgi and endoplasmic reticulum. Other inclusions that are absent in some other marsupial species are present in growing possum oocytes in smaller quantities, including yolklike lipid bodies and electron-dense granular bodies., (Copyright 2001 Wiley-Liss, Inc.)

Several morphological features of early development differ between marsupials and eutherians. These include early overt conceptus polarity, the mode of segregation of pluriblast and trophoblast lineages, and the superficial position of the pluriblast within the unilaminar blastocyst epithelium. Knowledge of the molecular mechanisms of early development is lacking in marsupials. We have cloned from the brushtail possum, Trichosurus vulpecula, the partial cDNA of a marsupial homologue of mammalian POU5F1, which encodes octamer-binding transcription factor-4 (Oct-4). The 579 bp cloned coding sequence of possum POU5F1 (tvPOU5F1) shares 74, 78, and 79% identity with murine Pou5f1, human POU5F1, and bovine bPOU5F1, respectively, at the protein level. The mRNA for tvPOU5F1 was detected by in situ hybridization in oogonia and in oocytes of growing follicles, but not in early meiotic oocytes. In a gastrulation-stage conceptus, high tvPOU5F1 expression was found in the embryonic ectoderm, with low or undetectable expression in other cell types. These results are comparable with those of murine and bovine expression studies, and suggest that the putative role of POU5F1 in totipotent and pluripotent cells is conserved among mammals., (Copyright 2001 Wiley-Liss, Inc.)

Modern concepts of gait rehabilitation after stroke favor a task-specific repetitive approach. In practice, the required physical effort of the therapists limits the realization of this approach. Therefore, a mechanized gait trainer enabling nonambulatory patients to have the repetitive practice of a gait-like movement without overstraining therapists was constructed. This preliminary study investigated whether an additional 4-week daily therapy on the gait trainer could improve gait ability in 14 chronic wheelchair-bound hemiparetic subjects. The 4 weeks of physiotherapy and gait-trainer therapy resulted in a relevant improvement of gait ability in all subjects. Velocity, cadence, and stride length improved significantly (p < 0.01). The kinesiologic electromyogram of selected lower-limb muscles revealed a more physiologic pattern. The confounding influence of spontaneous recovery, the lack of a control group, and the double amount of therapy limit the clinical relevance of this study. Nevertheless, the gait trainer seems feasible as an adjunctive tool in gait rehabilitation after stroke; further studies are needed.

In marsupials, the mechanisms of lineage allocation into pluriblast and trophoblast are related to conceptus polarity and polarized discharge of extracellular matrix (ECM). The brushtail possum, Trichosurus vulpecula, a major pest species in New Zealand, is being intensively studied to develop an immunocontraceptive control method. Of 23 specimens examined, 11 were examined by electron microscopy to study the presence and role of the ECM in lineage allocation in the possum. A number of polarized features in the zygote identified the future embryonic and abembryonic poles. Pronuclei, in a broad band of mitochondrion-rich cortical cytoplasm, lay in the embryonic hemisphere, and numerous electron-lucent vesicles characterized the abembryonic cytoplasm. These vesicles seemed to contribute to the ECM. During cleavage, cells lay near the zona in the embryonic hemisphere, and ECM accumulated chiefly in the abembryonic hemisphere. Cell-zona adhesion facilitated by microvillous and club processes occurred at the early 4-cell stage, and cell-cell adhesion commenced at the late 4-cell stage. The first two cleavages were meridional, equal, and accompanied by elimination into the cleavage cavity of much of the electron-lucent vesicular material in the form of several membrane-bound yolk masses. The third cleavage was unequal, with both meridional and latitudinal planes. The first differences between trophoblast and pluriblast lineages appeared at the 8-cell stage. Later cleavage planes were latitudinal or oblique. Conceptus polarity, polarized discharge of ECM, and localized cell-zona adhesion were related to the first lineage allocation in the possum.

The past decade has produced considerable debate over the feasibility of paleodemographic research, with much attention focusing on the question of reliability of age estimates. We show here that in cases where age is estimated rather than known, the traditional method of assigning individuals to age classes will produce biased estimates of age structure. We demonstrate the effect of this bias both mathematically and by computer simulation, and show how a more appropriate method from the fisheries literature (the "iterated age length key") can be used to estimate age structure. Because it is often the case that ages are also estimated for extant groups, we suggest that our results are relevant to the general field of anthropological demography, and that it is time for us to improve the statistical basis for age structure estimation. We further suggest that the oft noted paucity of older individuals in skeletal collections is a simple result of the use of inappropriate methods of age estimation, and that this problem can be rectified in the future by using maximum likelihood estimates of life table or hazard functions incorporating the uncertainty of age estimates.

Potential mates analysis is difficult to apply to small historic populations that lack clear boundaries or regular vital event registration. Here I analyze the actual mate pool as an alternative way to identify causes of nonrandom mating when unmarried members are unknown. Factors influencing mate choice within a historic eastern Blue Ridge community in Madison County, Virginia, are examined for four marriage cohorts: 1850-1879, 1880-1899, 1900-1919, and 1920-1939. These factors include nuclear kin avoidance, preferred age differences between mates, and preferences for more distant kin. A simulation is used to recombine members of the cohort-specific pools of married individuals to generate the probabilities of various types of kin marriages. The pedigree and vital statistics data are derived from first-time marriage licenses filled by community members in Madison County from 1794 to 1939. The numbers of marriages examined for each cohort are 88, 120, 132, and 132, respectively; the mate pools constructed from the samples are viewed from the female perspective. The results generated by simulation on the actual mate pools consist of mean kinship coefficients, numbers of marriages between "allowed" kin types, and probabilities of these values when marriage is random with respect to kinship. The results indicate significantly high levels of inbreeding in all four marriage cohorts, primarily because of high levels of first-cousin marriages in the first three cohorts and of first-cousin once-removed marriages in the 1920 cohort. The observed mating patterns are discussed in terms of the social history of the Blue Ridge community and restrictions of the data.

Biological distance analysis, the dominant type of skeletal biological research during the 19th century, has become less visible in recent years. Although the proportion of American Journal of Physical Anthropology articles and published abstracts focusing on biodistance has remained fairly constant over the three decades between 1955 and 1985, the proportion of biodistance contributions relative to other skeletal biology studies has decreased. Emphasis in skeletal biology has shifted from the analysis of biological variation to investigations of health and diet, and within biodistance studies methodological issues have assumed prominence over purely analytical approaches. This paper investigates trends in biological distance analysis through a survey of articles and meetings abstracts published in the American Journal of Physical Anthropology from 1955 to 1985. The survey provides the historical context for five symposium papers on skeletal biological distance presented at the 1986 meeting of the American Association of Physical Anthropologists.

The clinical efficacy of a new device for treatment of female incontinence was studied in a multicenter trial. The device consists of an inflatable electrode carrier and an external stimulator unit. Forty women were treated: 10 had primary or recurrent genuine stress incontinence, 15 had urge incontinence due to idiopathic detrusor instability, not responding to drug treatment, and 15 had stress incontinence combined with detrusor instability. Twenty-five patients were improved by the treatment. Another 8 reported an excellent result of treatment and remained free of symptoms for more than six months after withdrawal of the treatment. The results were more favorable in patients with bladder hyperactivity than in genuine stress incontinence. The patients' general ratings of treatment efficacy correlated well with their recordings of urinary frequency and consumption of incontinence pads. The functional bladder capacity increased in improved patients, but normalization of urodynamic parameters was no prerequisite for clinical improvement. We found intravaginal electrical stimulation to be a valuable alternative to medical and surgical intervention in patients with detrusor instability.

37 infants with brain tumours diagnosed in the first three years of life were studied. Among the 28 histologically verified tumours were 7 ependymomas, 6 medulloblastomas, 5 cerebellar astrocytomas, 3 astrocytomas and 2 spongioblastomas of the optic nerve. Features of clinical symptoms, therapy and follow-up studies are described. 7 infants survived in long-term follow-up with good or fair quality of life. Results of treatment were satisfactory in cerebellar astrocytomas only.