bluuebunny/crossref_metadata_2025 huggingface dataset:
Phorbol diester 12‐O‐tetradecanoylphorbol 13‐acetate (TPA) up‐regulates the expression of estrogen receptors in human THP‐1 leukemia cells
Authors: Cutolo Maurizio , Carruba Giuseppe , Villaggio Barbara , Coviello Domenico A. , Dayer Jean‐Michel , Campisi Ildegarda , Miele Monica , Stefano Rosalba , Castagnetta Luigi A.M.
Published: August 2002

In the present work, we have inspected expression of estrogen receptors (ER) and their regulation by the phorbol diester 12‐O‐tetradecanoylphorbol 13‐acetate (TPA) in a leukemic cell line, the THP‐1 cells, using multiple experimental approaches. Firstly, ligand binding assay (LBA) revealed that control (unstimulated) THP‐1 cells express type I (high affinity, limited capacity) ER in the nuclear fraction only, whilst treatment of cells with TPA resulted in the appearance of type I ER in the soluble fraction as well, with the 50 ng/ml dose and the 48 h incubation time being the most effective experimental condition. A concomitant increase of type II ER was also seen in both soluble and nuclear cell fractions. Unstimulated THP‐1 cells were found to be ER negative by immunocytochemistry; conversely, cells exposed to 50 ng/ml TPA for 48 h stained positively for ER, with the majority of cells having a strong nuclear staining. Scrutiny of ER mRNA expression using reverse transcriptase‐polymerase chain reaction showed the presence of a wild type ER transcript in both control and TPA‐treated THP‐1 cells, though levels of ER mRNA were found to be comparatively higher in the latter. This combined evidence would imply that the TPA‐induced differentiation of THP‐1 cells is accompanied by the rise of high affinity (type I) ER, suggesting that estrogens may play a role in the regulation of macrophage activity during the inflammatory and/or the immune response. J. Cell. Biochem. 83: 390–400, 2001. © 2001 Wiley‐Liss, Inc.
Free‐energy component analysis of 40 protein–DNA complexes: A consensus view on the thermodynamics of binding at the molecular level
Authors: Jayaram B. , McConnell K. , Dixit S. B. , Das A. , Beveridge D. L.
Published: August 2002

Noncovalent association of proteins to specific target sites on DNA—a process central to gene expression and regulation—has thus far proven to be idiosyncratic and elusive to generalizations on the nature of the driving forces. The spate of structural information on protein—DNA complexes sets the stage for theoretical investigations on the molecular thermodynamics of binding aimed at identifying forces responsible for specific macromolecular recognition. Computation of absolute binding free energies for systems of this complexity transiting from structural information is a stupendous task. Adopting some recent progresses in treating atomic level interactions in proteins and nucleic acids including solvent and salt effects, we have put together an energy component methodology cast in a phenomenological mode and amenable to systematic improvements and developed a computational first atlas of the free energy contributors to binding in ∼40 protein–DNA complexes representing a variety of structural motifs and functions. Illustrating vividly the compensatory nature of the free energy components contributing to the energetics of recognition for attaining optimal binding, our results highlight unambiguously the roles played by packing, electrostatics including hydrogen bonds, ion and water release (cavitation) in protein–DNA binding. Cavitation and van der Waals contributions without exception favor complexation. The electrostatics is marginally unfavorable in a consensus view. Basic residues on the protein contribute favorably to binding despite the desolvation expense. The electrostatics arising from the acidic and neutral residues proves unfavorable to binding. An enveloping mode of binding to short stretches of DNA makes for a strong unfavorable net electrostatics but a highly favorable van der Waals and cavitation contribution. Thus, noncovalent protein–DNA association is a system‐specific fine balancing act of these diverse competing forces. With the advances in computational methods as applied to macromolecular recognition, the challenge now seems to be to correlate the differential (initial vs. final) energetics to substituent effects in drug design and to move from affinity to specificity. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 1–14, 2002
Quasirelativistic study of 125Te nuclear magnetic shielding constants and chemical shifts
Authors: Hada Masahiko , Wan Jian , Fukuda Ryoichi , Nakatsuji Hiroshi
Published: August 2002

Calculations for 125Te magnetic shielding constants and chemical shifts were carried out using a quasirelativistic Hamiltonian including the spin‐free relativistic, one‐ and two‐electron spin–orbit, and relativistic magnetic interaction terms. For the tellurium‐containing series Te(CH3)2, TeH2, TeF6, Te(CH3)4, and Te(CH3)2Cl2, the relativistic effects amounted to as much as 1300 ppm and were very important for qualitatively reproducing the absolute value of the 125Te shielding constants obtained experimentally. On the other hand, for the 125Te chemical shifts the relativistic effects were less important, because they cancelled each other between the sample and reference compounds. © 2001 John Wiley & Sons, Inc. J Comput Chem 22: 1502–1508, 2001
Clinical implications of research on countertransference: Science informing practice
Authors: Hayes Jeffrey A. , Gelso Charles J.
Published: August 2002

This article presents a clinically oriented synthesis of research findings pertaining to countertransference (CT). Findings are organized around five therapy‐relevant questions concerning the origins, triggers, manifestations, management, and effects of CT. To date, research has focused primarily on impediments posed by therapists' CT reactions and mechanisms for managing them. It is argued that future research needs to be directed toward better understanding how therapists can use CT to enhance their work with clients. © 2001 John Wiley & Sons, Inc. J Clin Psychol/In Session 57: 1041–1051, 2001.
Role of AKT kinase in sphingosine‐induced apoptosis in human hepatoma cells
Authors: Chang Hui‐Chiu , Tsai Li‐Huang , Chuang Lea‐Yea , Hung Wen‐Chun
Published: August 2002

Our previous work has shown that a number of sphingolipid metabolites including sphingosine, sphinganine, and other long‐chain bases potently induced apoptosis in human hepatoma cells. In this study, we examined the possibility that sphingosine may trigger apoptosis in human hepatoma cells via inhibition of anti‐apoptotic pathways. We investigated the effect of sphingosine on AKT kinase, a serine/threonine kinase which was found to protect cells from apoptosis induced by a variety of extracellular stresses. Our results indicated that sphingosine inhibited basal and serum‐stimulated AKT kinase activity in a dose‐dependent manner in hepatoma cells. Additionally, sphingosine‐induced inhibition of AKT kinase was correlated with induction of apoptosis in these cells. Pretreatment of insulin, a potent stimulator of AKT kinase, partially reversed the inhibition of AKT kinase by sphingosine and counteracted the apoptotic action of this sphingolipid. Expression of activated AKT kinase partially protected cells from sphingosine‐induced apoptosis, whereas expression of kinase‐dead AKT kinase had no effect. The molecular mechanism by which AKT kinase suppressed the apoptotic action of sphingosine was investigated. Our results showed that increased release of cytochrome C from mitochondria and subsequent activation of caspase‐3 were detected in sphingosine‐treated hepatoma cells. On the contrary, expression of activated AKT kinase in Hep3B cells attenuated cytochrome C release and caspase‐3 activation induced by sphingosine. Taken together, these findings suggest that suppression of AKT kinase is one of the mechanisms by which sphingosine induces apoptosis in hepatoma cells and activation of AKT kinase may inhibit sphingosine‐induced apoptosis by blocking a step upstream of cytochrome C release and caspase‐3 activation. © 2001 Wiley‐Liss, Inc.
Phage‐mediated transfer of virulence genes
Authors: Saunders Jon R , Allison Heather , James Chloe E , McCarthy Alan J , Sharp Richard
Published: August 2002

Bacteriophages as accessory genetic elements play a crucial role in the dissemination of genes and the promotion of genetic diversity within bacterial populations. Such horizontal transfer of DNA is critical in the emergence of new pathogenic organisms, through the dissemination of genes encoding virulence factors such as toxins, adhesins and agressins. Phages can transfer genes that are not necessary for bacteriophage persistence and are generally recognised by their ability to convert their host bacteria to new phenotypes. This phenomenon is known as phage conversion. If such converting genes encode for virulence factors, the consequences of phage infection may include increased virulence of the host bacteria, and the conversion of a non‐pathogenic strain to a potentially dangerous pathogen. A number of virulence factors in bacteria causing diseases in plants, animals and humans are encoded by converting phages, the vast majority of which are temperate as opposed to lytic in nature.

© 2001 Society of Chemical Industry
Ultrasound‐guided testicular sperm aspiration in azoospermic patients: A new sperm retrieval method for intracytoplasmic sperm injection
Authors: Belenky Alexander , Avrech Ory M. , Bachar Gil Nissim , Zuckerman Zvi , Rafael Zion Ben , Fisch Benjamin , Cohen Maya
Published: August 2002

Purpose

We investigated the technique of ultrasound‐guided testicular sperm aspiration (USTSA) and compared it with “blind” testicular sperm aspiration (TSA) in patients with nonobstructive azoospermia.

Methods

Thirty‐nine consecutive azoospermic men underwent TSA, 16 under sonographic guidance (USTSA group) and 23 with no imaging guidance (TSA group). Clinical and hormonal evaluation and sonography of the scrotum and testes were performed 1–2 days before the procedure. The aspiration was done using short‐term general anesthesia. Follow‐up consisted of sonographic reexamination of the scrotum and testes immediately and 1 month after the procedure.

Results

Intraoperative sonography with power Doppler imaging enabled good visualization of the testicular parenchyma, easy sampling, and avoidance of prominent vessels. Sufficient material was retrieved in 15 USTSA patients (94%) and 19 TSA patients (83%). No patients needed more than 4 hours' ambulatory hospitalization after the procedure. In the remaining 5 patients, aspiration failed to yield sperm, so open biopsy was performed. In those patients, postaspiration surgical exploration revealed subtunical bleeding in 3 patients after TSA but none after USTSA. Late minor complications occurred in 2 patients (13%) in the USTSA group and 7 (30%) in the TSA group. No difference was found between the 2 groups in pregnancy rate in the patients' female partners.

Conclusions

USTSA is a safe and accurate method for sperm retrieval in azoospermic patients. © 2001 John Wiley & Sons, Inc. J Clin Ultrasound 29:339–343, 2001.
Adrenomedullin as a pancreatic hormone
Authors: López José , Cuesta Natalia
Published: August 2002

Adrenomedullin (AM) is a multiregulatory peptide which is expressed in a wide range of tissues. In the pancreas, AM was first found in mammals, including man, and its colocalization with the pancreatic polypeptide (PP) was established in islet F cells. In addition, three different AM receptors have been characterized in B‐cells. AM has been also located in the pancreatic cells of other vertebrate classes. The frequency and distribution of AM cells vary between different animals; they can be found scattered among the exocrine tissue, in the islets, or in ductal epithelia. The colocalization of AM with other hormones presents different patterns, although in birds, as in mammals, it seems to colocalize only with PP. The best‐determined pancreatic AM function is the inhibition of insulin secretion in B‐cells, which seems to be linked to a recently discovered binding protein, factor H. In relation to this physiological role, clinical data show that AM is raised in some groups of both types I and II diabetic patients and AM might have triggered the disease in a subset of them. On the other hand, AM pancreatic cells are also involved in the response to septic shock by increasing AM circulating levels. A third putative function is the inhibition of amylase secretion by the exocrine pancreatic cells. AM has been found in embryonic mammalian pancreas from the earliest stages of the development, colocalizing with all pancreatic hormones, although in adults only coexpression with PP is kept. AM may play a role in the growth and morphogenesis of the pancreas. Microsc. Res. Tech. 57:61–75, 2002. © 2002 Wiley‐Liss, Inc.
Role of cell cycle molecules in the pathophysiology of glomerular epithelial cells
Authors: Nagata Michio , Shu Yujing , Tomari Shinsuke
Published: August 2002

The postmitotic characteristics of podocytes are the basis for the structural assembly and central function (filtration) of the glomeruli. Persistent cell cycle quiescence is required for stability in these cells. In cells of the podocyte lineage, transdifferentiation from the metanephric mesenchyme to mature podocytes is closely associated with tight regulation of the expression of cell cycle molecules. This cell cycle control in podocytes acts as a safeguard for the glomeruli; however, deregulation of this system might result in structural deterioration. This article focuses on the expression of cell cycle molecules in podocyte differentiation and pathology. Microsc. Res. Tech. 57:203–207, 2002. © 2002 Wiley‐Liss, Inc.
Morphological and cytochemical aspects of capillary permeability
Authors: Bendayan Moise
Published: August 2002

Transport of plasma soluble constituents across the capillary wall is of primordial importance in cardiovascular physiology. While physiological experiments have concluded with the existence of two sets of pores, a large one responsible for the transport of proteins and a small one designed for the diffusion of small solutes, the morphological counterparts have yet to get general agreement. In this review, we present the different proposed paths within and between the endothelial cells that do allow passage of plasma constituents and may respond to the definitions established by physiological means. The vesicular system existing in endothelial cells has been the first transendothelial path to be proposed. Several data have demonstrated the involvement of this system in transport, although others have systematically brought controversy. One alternative to the vesicles has been the demonstration of membrane‐bound tubules creating, in certain cases, transendothelial channels that would allow diffusion of plasma proteins and other constituents across the capillary wall. Access to this tubulo‐vesicular system could be restrained by the stomatal diaphragm and facilitated by specific membrane receptors. Further, we have demonstrated for the first time with morpho‐cytochemical tools, that the intercellular clefts are the site of diffusion for small molecules such as peptides having a molecular weight inferior to 3,000. For the fenestrated capillary bed, we have shown that fenestrae are the site through which plasma constituents cross the capillary wall. However, and in spite of the existence of these large open pores, the endothelial cells still display the tubulo‐vesicular system involved in transport of large molecules and their intercellular clefts are also the site of diffusion of small molecules. Making consensus on the existence of an intracellular tubulo‐vesicular system in non‐fenestrated capillaries, responsible for the transport of large molecules by the endothelial cells, and understanding the rational for the fenestrated capillary to have three paths for transport—the fenestrae, the tubulo‐vesicular system, and the inter‐endothelial clefts—require further investigation. Microsc. Res. Tech. 57:327–349, 2002. © 2002 Wiley‐Liss, Inc.
Dynamics of Na+,K+,2Cl− cotransporter and Na+,K+‐ATPase expression in the branchial epithelium of brown trout (Salmo trutta) and atlantic salmon (Salmo salar)
Authors: Tipsmark Christian Kølbæk , Madsen Steffen Søndergaard , Seidelin Michel , Christensen Akim Stypinsky , Cutler Christopher Paul , Cramb Gordon
Published: August 2002

The dynamics of branchial Na+,K+,2Cl− cotransporter (NKCC) and Na+,K+‐ATPase (NKA) expression were investigated in brown trout and Atlantic salmon during salinity shifts and the parr‐smolt transformation, respectively. In the brown trout, Western blotting revealed that NKCC and NKA abundance increased gradually and in parallel (30‐ and ten‐fold, respectively) after transfer to seawater (SW). The NKA hydrolytic activity increased ten‐fold after SW‐transfer. Following back‐transfer to fresh water (FW), the levels of both proteins and NKA activity decreased. The NKCC immunostaining in the gill of SW‐acclimated trout was strong, and mainly localized in large cells in the filament and around the bases of the lamellae. In FW‐acclimated trout, immunostaining was less intense and more diffuse. Partial cDNAs of the secretory NKCC1 isoform were cloned and sequenced from both brown trout and Atlantic salmon gills. Two differently sized transcripts were detected by Northern blotting in the gill but not in other osmoregulatory tissues (kidney, pyloric caeca, intestine). The abundance in the gill of these transcripts and of the associated NKCC protein increased four‐ and 30‐fold, respectively, during parr‐smolt transformation. The abundance of NKA α‐subunit protein also increased in the gill during parr‐smolt transformation though to a lesser extent than enzymatic activity (2.5‐ and eight‐fold, respectively). In separate series of in vitro experiments, cortisol directly stimulated the expression of NKCC mRNA in gill tissue of both salmonids. The study demonstrates the coordinated regulation of NKCC and NKA proteins in the gill during salinity shifts and parr‐smolt transformation of salmonids. © 2002 Wiley‐Liss, Inc.
Head shape and bite performance in xenosaurid lizards
Authors: Herrel Anthony , De Grauw Ed , Lemos‐Espinal Julio A.
Published: August 2002

Bite performance in lizards influences many aspects of the animal’s lifestyle. During feeding, inter‐ and intrasexual interactions, and defensive behavior, the ability to bite hard might be advantageous. Although biomechanical considerations predict clear relations between head shape and bite performance, this has rarely been tested. Here we investigate the effect of head shape on bite performance in three closely related species of xenosaurid lizards. Our data show that in this family of lizards, bite performance is mainly determined by head height, with high headed animals biting harder than flat headed ones. Species clearly differ in head shape and bite performance and show a marked sexual dimorphism. The dimorphism in head shape also results in an intersexual difference in bite performance. As head height is the major determinant of bite performance in xenosaurid lizards, trade offs between a crevice dwelling life‐style and bite performance seem to occur. The evolutionary implications of these results are discussed. J. Exp. Zool. 290:101–107, 2001. © 2001 Wiley‐Liss, Inc.
Relationship between cerebro‐spinal fluid pH and pulmonary ventilation of the South American lungfish, Lepidosiren paradoxa (Fitz.)
Authors: Sanchez Adriana Paula , Hoffmann Anette , Rantin Francisco Tadeu , Glass Mogens Lesner
Published: August 2002

The respiratory control in land vertebrates (Tetrapoda) is mainly linked to regulation of acid–base status, which involves peripheral and central chemoreceptors. The lungfish (Dipnoi) might constitute the sister group of all land vertebrates (Tetrapoda) and possess a combination of real lungs and reduced gills. In this context, we evaluated the possible presence of central respiratory chemoreceptors in the South American Lungfish, Lepidosiren paradoxa. Pulmonary ventilation and respiratory frequency increased significantly with reductions of CSF pH by means of mock CSF solutions. This suggests that Lepidosiren possess central acid–base receptors. J. Exp. Zool. 290:421–425, 2001. © 2001 Wiley‐Liss, Inc.
Wnts and the female reproductive system
Authors: Heikkilä Minna , Peltoketo Hellevi , Vainio Seppo
Published: August 2002

Wnts are intercellular growth and differentiation factors that regulate several key developmental steps, such as gastrulation, neurulation, and organogenesis, including the development of the midbrain, central nervous system, kidney, and limbs. Wnts are also needed for a normal development of the reproductive system. Deficiency of Wnt‐4, ‐5a, and ‐7a, for example, results in sex reversal, infertility, and/or malformation of the internal and external genitals. Here we focus on the importance of Wnts in the female reproductive system. J. Exp. Zool. 290:616–623, 2001. © 2001 Wiley‐Liss, Inc.
Combinatorial curvature for planar graphs
Authors: Higuchi Yusuke
Published: August 2002

Regarding an infinite planar graph G as a discrete analogue of a noncompact simply connected Riemannian surface, we introduce the combinatorial curvature of G corresponding to the sectional curvature of a manifold. We show this curvature has the property that its negative values are bounded above by a universal negative constant. We also prove that G is hyperbolic if its curvature is negative. © 2001 John Wiley & Sons, Inc. J Graph Theory 38: 220–229, 2001
Sequence Heterochrony and the Evolution of Development
Authors: Smith Kathleen K.
Published: August 2002

One of the most persistent questions in comparative developmental biology concerns whether there are general rules by which ontogeny and phylogeny are related. Answering this question requires conceptual and analytic approaches that allow biologists to examine a wide range of developmental events in well‐structured phylogenetic contexts. For evolutionary biologists, one of the most dominant approaches to comparative developmental biology has centered around the concept of heterochrony. However, in recent years the focus of studies of heterochrony largely has been limited to one aspect, changes in size and shape. I argue that this focus has restricted the kinds of questions that have been asked about the patterns of developmental change in phylogeny, which has narrowed our ability to address some of the most fundamental questions about development and evolution. Here I contrast the approaches of growth heterochrony with a broader view of heterochrony that concentrates on changes in developmental sequence. I discuss a general approach to sequence heterochrony and summarize newly emerging methods to analyze a variety of kinds of developmental change in explicit phylogenetic contexts. Finally, I summarize a series of studies on the evolution of development in mammals that use these new approaches. J. Morphol. 252:82–97, 2002. © 2002 Wiley‐Liss, Inc.
Evolution of body‐wall musculature in the Platyhelminthes (Acoelomorpha, Catenulida, Rhabditophora)
Authors: Hooge Matthew D.
Published: August 2002

In an effort to understand the phylogeny of the Platyhelminthes, the patterns of body‐wall musculature of flatworms were studied using fluorescence microscopy and Alexa‐488‐labeled phalloidin. Species of the Catenulida have a simple orthogonal gridwork of longitudinal and circular muscles. Members of the Rhabditophora have the same gridwork of musculature, but also have diagonal muscles over their entire body. Although a few species of Acoelomorpha possessed a simple orthogonal grid of musculature, most species typically have distinctly different patterns of dorsal and ventral body‐wall musculature that include sets of longitudinal, circular, U‐shaped, and several kinds of diagonal muscles. Several distinct patterns of musculature were identified, including 8 patterns in 11 families of acoels. These patterns have proven to be useful in clarifying the phylogeny of the Acoelomorpha, particularly with regard to the higher acoels. Patterns of musculature as well as other morphological characters are used here for revisions of acoel systematics, including the return of Eumecynostomum sanguineum (Mecynostomidae) to the genus Aphanostoma (Convolutidae), the revision of the family Childiidae, and the formation of a new family, Actinoposthiidae. J. Morphol. 249:171–194, 2001. © 2001 Wiley‐Liss, Inc.
Ultrastructural evidence for two‐cell and three‐cell neural pathways in the tentacle epidermis of the sea anemone Aiptasia pallida
Authors: Westfall Jane A. , Elliott Carol F. , Carlin Ryan W.
Published: August 2002

Sensory and ganglion cells in the tentacle epidermis of the sea anemone Aiptasia pallida were traced in serial transmission electron micrographs to their synaptic contacts on other cells. Sensory cell synapses were found on spirocytes, muscle cells, and ganglion cells. Ganglion cells, in turn, synapsed on sensory cells, spirocytes, muscle cells, and other neurons and formed en passant axo‐axonal synapses. Axonal synapses on nematocytes and gland cells were not traced to their cells of origin, i.e., identified sensory or ganglion cells. Direct synaptic contacts of sensory cells with spirocytes and sensory cells with muscle cells suggest a local two‐cell pathway for spirocyst discharge and muscle cell contraction, whereas interjection of a ganglion cell between the sensory and effector cells creates a local three‐cell pathway. The network of ganglion cells and their processes allows for a through‐conduction system that is interconnected by chemical synapses. Although the sea anemone nervous system is more complex than that of Hydra, it has similar two‐cell and three‐cell effector pathways that may function in local responses to tentacle contact with food. J. Morphol. 251:83–92, 2002. © 2002 Wiley‐Liss, Inc.
Fine structure and immunocytochemistry of a new chemosensory system in the chiton larva (Mollusca: Polyplacophora)
Authors: Haszprunar Gerhard , Friedrich Stefan , Wanninger Andreas , Ruthensteiner Bernhard
Published: August 2002

Combined electron microscopy and immunocytochemistry of the larvae of several polyplacophoran species (Chiton olivaceus, Lepidochitona aff. corrugata, Mopalia muscosa) revealed a sensory system new to science, a so‐called “ampullary system.” The cells of the “ampullary system” are arranged in four symmetrically situated pairs lying dorsolaterally and ventrolaterally in the pretrochal part of the trochophore‐like larva and they send axons into the cerebral commissure. They are lost at metamorphosis. The fine structure of these cells strongly resembles that of so‐called “ampullary cells” known from various sensory organs of other molluscs, such as the apical complex of gastropod and bivalve larvae, osphradia of vetigastropods, and olfactory organs of cephalopods, and nuchal organs of certain polychaetes. The ampullary cells and their nerves are densely stained by anti‐FMRF‐amide fluorescence dyes, whereas antiserotonin staining is only weak. While cytological homology of the ampullary cells with those of other organs is probable, the ampullary system as a whole is regarded as a synapomorphy of the Polyplacophora or Chitonida. J. Morphol. 251:210–218, 2002. © 2002 Wiley‐Liss, Inc.
Ontogenetic and phylogenetic transformations of the ear ossicles in marsupial mammals
Authors: Sánchez‐Villagra Marcelo R. , Gemballa Sven , Nummela Sirpa , Smith Kathleen K. , Maier Wolfgang
Published: August 2002

This study is based on the examination of histological sections of specimens of different ages and of adult ossicles from macerated skulls representing a wide range of taxa and aims at addressing several issues concerning the evolution of the ear ossicles in marsupials. Three‐dimensional reconstructions of the ear ossicles based on histological series were done for one or more stages of Monodelphis domestica, Caluromys philander, Sminthopsis virginiae, Trichosurus vulpecula, and Macropus rufogriseus. Several common trends were found. Portions of the ossicles that are phylogenetically older develop earlier than portions representing more recent evolutionary inventions (manubrium of the malleus, crus longum of the incus). The onset of endochondral ossification in the taxa in which this was examined followed the sequence; first malleus, then incus, and finally stapes. In M. domestica and C. philander at birth the yet precartilaginous ossicles form a supportive strut between the lower jaw and the braincase. The cartilage of Paauw develops relatively late in comparison with the ear ossicles and in close association to the tendon of the stapedial muscle. A feeble artery traverses the stapedial foramen of the stapes in the youngest stages of M. domestica, C. philander, and Sminthopsis virginiae examined. Presence of a large stapedial foramen is reconstructed in the groundplan of the Didelphidae and of Marsupialia. The stapedial foramen is absent in all adult caenolestids, dasyurids, Myrmecobius, Notoryctes, peramelids, vombatids, and phascolarctids. Pouch young of Perameles sp. and Dasyurus viverrinus show a bicrurate stapes with a sizeable stapedial foramen. Some didelphids examined to date show a double insertion of the Tensor tympani muscle. Some differences exist between M. domestica and C. philander in adult ossicle form, including the relative length of the incudal crus breve and of the stapes. Several differences exist between the malleus of didelphids and that of some phalangeriforms, the latter showing a short neck, absence of the lamina, and a ventrally directed manubrium. Hearing starts in M. domestica at an age in which the external auditory meatus has not yet fully developed, the ossicles are not fully ossified, and the middle ear space is partially filled with loose mesenchyme. The ontogenetic changes in hearing abilities in M. domestica between postnatal days 30 and 40 may be at least partially related to changes in middle ear structures. J. Morphol. 251:219–238, 2002. © 2002 Wiley‐Liss, Inc.
Ultrastructure of the reproductive system of the black swamp snake (seminatrix pygaea). III. Sexual segment of the male kidney
Authors: Sever David M. , Stevens Rebecca A. , Ryan Travis J. , Hamlett William C.
Published: August 2002

In mature male snakes and lizards, a distal portion of the nephron is hypertrophied in relation to its appearance in females and immature males. This sexual segment of the male kidney apparently provides seminal fluid that is mixed with sperm and released into the female cloaca during copulation. In this article, we provide the first study at the ultrastructural level of seasonal variation in the sexual segment of the kidney of a squamate, the natricine snake Seminatrix pygaea. Previous workers have indicated that the sexual segment is secretory only when the testes are spermatogenically active. The sexual segment of the kidney in S. pygaea does not go through an extended period of inactivity but does show a cycle of synthesis and secretion that can be related to the spermatogenic cycle and mating activity. We show that synthesis of secretory product is initiated with the onset of spermatogenic activity in the spring and culminates with completion of spermiation in the fall. Secretion of the product, however, occurs in a premating period in March when the testes are inactive. Secretion during this premating period is probably necessary to provide time for the passage of the products down the ureter in order to mix with sperm during mating later in spring. J. Morphol. 252:238–254, 2002. © 2002 Wiley‐Liss, Inc.
Comparative morphological assessment of the psyllid pleuron (insecta, hemiptera, sternorrhyncha)
Authors: Ouvrard David , Bourgoin Thierry , Campbell Bruce C.
Published: August 2002

A unique set of morphological characters based on the hemipteran (sensu lato) thorax are used to define the pleuron of Psylloidea. New external and internal topographical descriptions of pleurites of all three thoracic segments are provided based on observations of specimens from various genera representing the current taxonomic arrangement of Psylloidea. Variations in propleurite morphology and the anteroventral angle of the episternum among taxa are clarified. The mesothoracic pleural sulcus is found to be a distinct groove formed by the deep fossa of the pleural apophysis and is not a secondary structure, as assigned by previous authors. A newly discovered internal apodeme of the metathoracic trochantin and a serially homologous structure in the mesothorax isolate the trochantinal territory of the second segment. The metathoracic pleural sulcus appears to be pressed against the ventral edge of the metepimeron, as previously described for the mesothorax of certain species of other insects having a strongly developed meron. Use of morphological interpretations and newly discovered apodemes to assess primary homology of structures for phylogenetic and taxonomic studies is discussed. A glossary of standardized morphological terms for thoracic structures of non‐heteropteran Hemiptera based on this study and other recent and former dissertations on the insect thorax is provided. J. Morphol. 252:276–290, 2002. © 2002 Wiley‐Liss, Inc.
Functional morphology of the forelimb of Tupaiids (Mammalia, Scandentia) and its phylogenetic implications
Authors: Sargis Eric J.
Published: August 2002

In this study, the forelimb of 12 species of tupaiids was analyzed functionally and compared to that of other archontan mammals. Several differences that relate to differential substrate use were found in the forelimb morphology of tupaiids. These differences included shape of the scapula, length and orientation of the coracoid process, size of the lesser tuberosity, shape of the capitulum, length of the olecranon process, and shape of the radial head and central fossa. The forelimb of the arboreal Ptilocercus lowii, the only ptilocercine, is better adapted for arboreal locomotion, while that of tupaiines is better adapted for terrestrial (or scansorial) locomotion. While the forelimb of the arboreal Ptilocercus appears to be habitually flexed and exhibits more mobility in its joints, a necessity for movement on uneven, discontinuous arboreal supports, all tupaiines are characterized by more extended forelimbs and less mobility in their joints. These restricted joints limit movements more to the parasagittal plane, which increases the efficiency of locomotion on a more even and continuous surface like the ground. Even the most arboreal tupaiines remain similar to their terrestrial relatives in their forelimb morphology, which probably reflects the terrestrial ancestry of Tupaiinae (but not Tupaiidae). The forelimb of Urogale everetti is unique among tupaiines in that it exhibits adaptations for scratch‐digging. Several features of the tupaiid forelimb reflect the arboreal ancestry of Tupaiidae and it is proposed that the ancestral tupaiid was arboreal like Ptilocercus. Also, compared to the forelimb character states of tupaiines, those of Ptilocercus are more similar to those of other archontans and it is proposed that the attributes of the forelimb of Ptilocercus are primitive for the Tupaiidae. Hence, Ptilocercus should be considered in any phylogenetic analysis that includes Scandentia. J. Morphol. 253:10–42, 2002. © 2002 Wiley‐Liss, Inc.
Skull osteology of Parvilacerta parva, a small‐sized lacertid lizard from Asia Minor
Authors: Müller Johannes
Published: August 2002

The skull of the poorly known Asia Minor lacertid lizard Parvilacerta parva is described. In comparison to other palaearctic lacertids, Parvilacerta parva displays several peculiar traits such as a large braincase, a short parietal table, closed post‐temporal openings, reduced ossification, or a large lacrimal. Many features are related to the small size of Parvilacerta parva and are of apparently heterochronic nature, as they can also be found in certain ontogenetic stages of other lacertid species. Small size and heterochrony even seem to be factors involved in the less‐developed sexual dimorphism of Parvilacerta parva. The described morphology is interpreted to be related to a life history strategy in r‐selecting, semiarid environments. J. Morphol. 253:43–50, 2002. © 2002 Wiley‐Liss, Inc.
Focal liver lesions: Comparison of dual‐phase CT and multisequence multiplanar MR imaging including dynamic gadolinium enhancement
Authors: Semelka Richard C. , Martin Diego R. , Balci Cem , Lance Trang
Published: August 2002

The purpose of this study was to compare dual‐phase spiral computed tomography (CT) and magnetic resonance imaging (MRI) using dynamic gadolinium enhancement for liver lesion detection and characterization. Twenty‐two consecutive patients underwent dual‐phase spiral CT and MRI for the evaluation of focal liver disease within a 1‐month period. Spiral CT and MR images were interpreted prospectively, in a blinded fashion by separate, individual, experienced investigators, to determine lesion detection and characterization. Liver lesions were confirmed by surgery and pathology in 6 patients, and by clinical and imaging follow‐up in the other 16 patients. Pathological correlation of a primary extrahepatic malignancy was available in 5 of the 16 patients who had metastatic liver disease. Spiral CT and MRI detected 53 and 63 lesions, and characterized 39 and 62 true positive lesions, respectively. A kappa statistic test was applied to assess agreement between MR and CT results. MR versus CT for lesion detection resulted in a kappa statistic of 0.54 (95% confidence interval), indicating moderate agreement, and 0.32 (95% confidence interval) for lesion characterization, indicating only slight agreement. More lesions were detected on MR images than CT images in 6 (27%) patients, with lesions detected only on MR images in 4 (18%) patients. More lesions were characterized on MR images in 9 (41%) patients. In 9 patients with a discrepancy between MR and CT findings, the MR images added information considered significant to patient management in all 9 cases. MRI was moderately superior to dual‐phase spiral CT for lesion detection, and was markedly superior for lesion characterization, with these differences having clinical significance. J. Magn. Reson. Imaging 2001;13:397–401. © 2001 Wiley‐Liss, Inc.
Identification of adulteration in milk by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Authors: Cozzolino R. , Passalacqua S. , Salemi S. , Malvagna P. , Spina E. , Garozzo D.
Published: August 2002

The development is described of a rapid, simply and accurate analytical method aimed at evaluating both the presence of cow milk in either raw ewe and water buffalo milk samples employed in industrial processes and the addition of powdered milk to samples of fresh raw milk, using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS). The presence of adulteration is defined by evaluating the protein patterns coming from the most abundant whey proteins, α‐lactalbumin and β‐lactoglobulin, used as molecular markers. As no pretreatment of the milk samples is required and owing to the speed and ease of use of MALDI‐MS the proposed analytical protocol can be used as a routine strategy for the identification of possible adulteration of the raw fresh milk samples that the dairy industry receives from producers every day. Copyright © 2001 John Wiley & Sons, Ltd.
Charge‐reduced nano electrospray ionization combined with differential mobility analysis of peptides, proteins, glycoproteins, noncovalent protein complexes and viruses
Authors: Bacher Gerold , Szymanski Wladyslaw W. , Kaufman Stanley L. , Zöllner Peter , Blaas Dieter , Allmaier Günter
Published: August 2002

This study explores the potential of a novel electrospray‐based method, termed gas‐phase electrophoretic mobility molecular analysis (GEMMA), allowing the molecular mass determination of peptides, proteins and noncovalent biocomplexes up to 2 MDa (dimer of immunglobulin M). The macromolecular ions were formed by nano electrospray ionization (ESI) in the ‘cone jet’ mode. The multiple charged state of the monodisperse droplets/ions generated was reduced by means of bipolar ionized air (generated by an α‐particle source) to yield exclusively singly charged positive and negative ions as well as neutrals. These ions are separated subsequently at atmospheric pressure using a nano differential mobility analyzer according to their electrophoretic mobility in air. Finally, the ions are detected using a standard condensation particle counter. Data were expressed as electrophoretic mobility diameters by applying the Millikan equation. The measured electrophoretic mobility diameters, or Millikan diameters, of 32 well‐defined proteins were plotted against their molecular weights in the range 3.5 to 1920 kDa and exhibited an excellent squared correlation coefficient (r2 = 0.999). This finding allowed the exact molecular weight determination of large (glyco)proteins and noncovalent biocomplexes by means of this new technique with a mass accuracy of ±5.6% up to 2 MDa at the femtomole level. From the molecular masses of the weakly bound, large protein complexes thus obtained, the binding stoichiometry of the intact complex and the complex stability as a function of pH, for example, can be derived. Examples of specific protein complexes, such as the avidin or catalase homo‐tetramer, are used to illustrate the potential of the technique for characterization of high‐mass biospecific complexes. A discussion of current and future applications of charge‐reduced nano ESI GEMMA, such as chemical reaction monitoring (reduction process of immunglobulin G) or size determination of an intact virus, a supramolecular complex, and monitoring of partial dissociation of a human rhinoviruses, is provided. Copyright © 2001 John Wiley & Sons, Ltd.
Symptom pathogenesis during acute influenza: Interleukin‐6 and Other cytokine responses
Authors: Kaiser Laurent , Fritz R. Scott , Straus Stephen E. , Gubareva Larisa , Hayden Frederick G.
Published: August 2002

In experimental human influenza infection initiated by nasal inoculation, the magnitude of viral replication, fever, and symptoms correlate with nasopharyngeal lavage fluid levels of various cytokines. Our aim was to assess these relationships in patients with naturally occurring acute influenza. Patients with culture‐positive influenza illness of less than 36 hr of duration were studied. Nasopharyngeal washing were collected at enrollment and on Day 2, 4, 6 and 8 for quantitative virus isolation and IL‐6, TNF‐α, INF‐α, INF‐γ and IL‐10 determinations. Blood samples collected at entry and on Day 2 and 6 were processed to assess plasma cytokines and circulating influenza RNA. Patients received either oseltamivir or placebo for 5 days. We assessed the correlation between nasopharyngeal lavage fluid or blood levels of cytokines before treatment and viral titers, symptom severity and fever. Sixteen adult subjects (median age of 22 years) were studied. In this small group of patients no significant differences between placebo and oseltamivir patients were found in viral replication or measures of cytokines. Thus the data for all 16 subjects were pooled for analysis. At entry, influenza A viruses were cultured from nasopharyngeal washes at a median titer of 4.8 log10TCID50/ml of wash. Viral titers correlated positively with symptom score (P = 0.006) and temperature values (P < 0.001). Viral titers, fever and symptoms were highest at enrollment and fell in parallel during the subsequent days. RT‐PCR assays failed to detect influenza RNA in the white blood cells from any patient. We observed a significant release, in both nasopharyngeal lavage fluid and in plasma, of IL‐6, TNF‐α, INF‐α, INF‐γ and IL‐10. At entry high IL‐6 levels were detected in the nasopharyngeal lavage fluid (median 10.3 pg/ml) and plasma (median 5.1 pg/ml) of all patients. We found a positive correlation between plasma IL‐6 levels and both symptom scores and temperature values (P < 0.05), as well as a positive correlation between nasopharyngeal lavage fluid levels of IL‐6 and TNF‐α and temperature (P < 0.05). We did not find significant associations between symptoms, fever and levels of INF‐α, INF‐γ or IL‐10. The magnitude of early decrease in viral titers correlated with initial levels of INF‐γ in nasopharyngeal lavage fluid (P < 0.05). Significant production of IL‐6, TNF‐α, INF‐α, INF‐γ and IL‐10 occurs in response to community acquired influenza A illness. As in experimental influenza, symptoms and fever in natural acute influenza correlate with the release of IL‐6. J. Med. Virol. 64:262–268, 2001. © 2001 Wiley‐Liss, Inc.
Detection of multiple human papillomavirus types in the lower genital tract correlates with cervical dysplasia***
Authors: Fife Kenneth H. , Cramer Harvey M. , Schroeder Jill M. , Brown Darron R.
Published: August 2002

Some human papillomavirus (HPV) types, such as HPV 16, are clearly associated with cervical dysplasia; however, the role played by other HPV types occasionally found in dysplasia is less certain. In addition, most methods used to detect HPV in clinical specimens cannot easily distinguish among more than two or three HPV types in a single specimen. Therefore, the significance of infection with multiple HPV types is not known. To address this question, we analyzed cervicovaginal lavage specimens from three cohorts of women for HPV DNA using a PCR/reverse blot assay system that permits the detection and partial quantitation of 26 genital HPV types. As expected, 94.1% of women who had dysplasia (n = 34) and 71.4% of women who had atypical squamous cells of uncertain significance (ASCUS) (n = 21) on cytology had HPV DNA detected compared to 54.5% of age matched women with normal cytology. HPV 16 DNA was detected in 35% of dysplasia patients compared to 9% of cytologic normals (P = 0.0044). Dysplasia patients had a mean of 3.29 (range 0–10) different HPV types detected compared to 1.04 (range 0–7) HPV types among those with normal cytology (P < 0.0001). These data support a possible role for multiple HPV types in the development or progression of cervical dysplasia. J. Med. Virol. 64:550–559, 2001. © 2001 Wiley‐Liss, Inc.
Detection of adeno‐associated virus DNA in female genital samples by PCR‐ELISA
Authors: Venturoli S. , Cricca M. , Bonvicini F. , Gallinella G. , Gentilomi G. , Zerbini M. , Musiani M.
Published: August 2002

Adeno‐associated viruses (AAV) are human parvoviruses that require helper function for their replication. Several studies have demonstrated that AAV DNA sequences can be found in the female genital tract but the incidence of infection seems very variable. A PCR‐ELISA method detecting AAV DNA was developed for combining the specificity and the sensitivity of conventional PCR with an objective interpretation of the results. In the PCR‐ELISA, a defined number of cells from cervical specimens were digested and amplified with concomitant digoxigenin labeling. Digoxigenin‐labeled amplified products hybridized to a specific biotinylated probe were captured in streptavidin‐coated microtiter wells by a biotin‐streptavidin binding and were visualized by colorimetric immunoenzymatic reaction. PCR‐ELISA was carried out in 110 cervical cytological specimens of women with or without the concomitant detectable presence of papillomavirus (HPV) DNA and the results were compared with those obtained by conventional PCR followed by dot blot hybridization. When compared to conventional PCR considered as reference standard, PCR‐ELISA was found to be 98% sensitive and 96% specific. Out of the total 110 samples examined, 52.7% were positive for AAV DNA by both techniques, demonstrating a high prevalence of AAV infection in the uterine cervix. When analyzing samples with or without the presence of HPV DNA, 63.2 % of the samples were positive for HPV DNA and 41.5% of the samples were negative for HPV proved positive for AAV DNA by both PCR‐ELISA and conventional PCR. Hence, PCR‐ELISA, which can be completed in 1 day, proved to be a reliable method for an objective detection of AAV DNA in clinical samples. The present study showed a frequent infection of the cervical epithelium with AAV both in the presence and absence of HPV infection. J. Med. Virol. 64:577–582, 2001. © 2001 Wiley‐Liss, Inc.