bluuebunny/crossref_metadata_2025 huggingface dataset:
Microtubule Organization and Function in Epithelial Cells
Authors: Müsch Anne
Published: November 2004

Microtubules are essential for many aspects of polarity in multicellular organisms, ranging from the asymmetric distribution of cell‐fate determinants in the one‐cell embryo to the transient polarity generated in migrating fibroblasts. Epithelial cells exhibit permanent cell polarity characterized by apical and basolateral surface domains of distinct protein and lipid composition that are segregated by tight junctions. They are also endowed with a microtubule network that reflects the asymmetry of their cell surface: microtubule minus‐ends face the apical‐ and microtubule plus‐ends the basal domain. Strikingly, the formation of distinct surface domains during epithelial differentiation is accompanied by the re‐organization of microtubules from a uniform array focused at the centrosome to the noncentrosomal network that aligns along the apico‐basolateral polarity axis. The significance of this coincidence for epithelial morphogenesis and the signaling mechanisms that drive microtubule repolymerization in developing epithelia remain major unresolved questions that we are only beginning to address. Studies in cultured polarized epithelial cells have established that microtubules serve as tracks that facilitate targeted vesicular transport. Novel findings suggest, moreover, that microtubule‐based transport promotes protein sorting, and even the generation of transport carriers in the endo‐ and exocytic pathways.
Stimulation‐Dependent Recycling of Integrin β1 Regulated by ARF6 and Rab11
Authors: Powelka Aimee M. , Sun Jianlan , Li Jian , Gao Minggeng , Shaw Leslie M. , Sonnenberg Arnoud , Hsu Victor W.
Published: November 2004

In comparison to the internalization pathways of endocytosis, the recycling pathways are less understood. Even less defined is the process of regulated recycling, as few examples exist and their underlying mechanisms remain to be clarified. In this study, we examine the endocytic recycling of integrin β1, a process that has been suggested to play an important role during cell motility by mediating the redistribution of integrins to the migrating front. External stimulation regulates the endocytic itinerary of β1, mainly at an internal compartment that is likely to be a subset of the recycling endosomes. This stimulation‐dependent recycling is regulated by ARF6 and Rab11, and also requires the actin cytoskeleton in an ARF6‐dependent manner. Consistent with these observations being relevant for cell motility, mutant forms of ARF6 that affect either actin rearrangement or recycling inhibit the motility of a breast cancer cell line.
Antibodies to Chlamydia trachomatis heat shock proteins Hsp60 and Hsp10 and subfertility in general population at age 31
Authors: Karinen L. , Pouta A. , Hartikainen A.‐L. , Bloigu A. , Paldanius M. , Leinonen M. , Saikku P. , Järvelin M.‐R.
Published: November 2004

Problem:  To assess the association between antibodies to Chlamydia trachomatis heat shock proteins 60 and 10 (Hsp60 and Hsp10) and subfertility in a general population sample.

Method of Study:  A nested case (n = 146)–control (n = 278) study in a population‐based birth cohort. Serum immunoglobulin (Ig)G and IgA antibodies against C. trachomatis Hsp60 and Hsp10, explanatory factors, were measured by enzyme immunoassay, using recombinant proteins as antigens. The main outcome variable was subfertility (time to pregnancy ≥12 months).

Results:  The prevalence and medians of serum IgA antibodies to Hsp60 and Hsp10 were significantly higher in the female partners of subfertile couple than in their fertile controls. On the contrary, among male partners of subfertile couple, especially among smokers serum antibody levels to Hsp antigens were lower than in the controls.

Conclusion:  The results indicate a serological association of antibodies to chlamydial Hsp antigens with female subfertility in a population‐based sample.
Identification of the M‐CSF Receptor in Endometriosis by Immunohistochemistry and RT‐PCR
Authors: Mettler Liselotte , Schmutzler Andreas G , Koch Kerstin , Schollmeyer Thoralf , Salmassi Ali
Published: November 2004

Problem:  The aim of this paper is to provide further evidence that the dystopic proliferation of endometriotic epithelia is caused by the stimulation of peritoneal macrophages. It is essential to show that endometriotic epithelial cells express the macrophage colony‐stimulating factor receptor (M‐CSFR) which binds the M‐CSF produced by the peritoneal macrophages.

Method of study:  For the detection of M‐CSFR, samples of ectopic endometrium (n = 79) and eutopic endometrium (n = 18) were compared. The specimens were gained at operative laparoscopy in the proliferative phase of the cycle. Cryostat sections were used for immunohistochemical detection. For in vitro reverse transcriptase polymerase chain reaction (RT‐PCR) tests, the tissue was immediately shock frozen on paraffin sections. For the in situ RT‐PCR technique the specimens were placed in a para‐formaldehyde solution, embedded in paraffin and later processed. The Gene Amp 1000 in situ PCR system (Perkin Elmer) was used as the thermal cycler.

Results:  M‐CSF and the M‐CSF receptor are present in eutopic and ectopic endometrium. Qualitatively, with both PCR techniques we found the M‐CSF receptor to be present in all samples examined. Using the histochemical detection technique, the M‐CSF receptor was found in nearly 70% of endometriosis patients compared with a statistically significant lower percentage in normal endometrium.

Conclusions:  The in situ RT‐PCR technique and immunohistochemistry elaborated the need to trace the cellular sources of the M‐CSF receptor. The identification of the M‐CSF receptor in endometriotic tissue and in endometrium is apt to open a new experimental field in endometriosis research.
Assessment of periodontal knowledge and periodontal status of an adult population in Jordan
Authors: El‐Qaderi SS , Quteish Ta'ani D
Published: November 2004

Abstract:  The aim of this study was to assess periodontal knowledge and periodontal status in 20–60‐year‐old adult population in northern Jordan. A convenient sample consisting of 722 adults was included in this study. A questionnaire incorporating items related to socio‐demographic and periodontal knowledge questions was completed before clinical examination, using the Community Periodontal Index of Treatment Needs (CPITN). The results showed that the proportions of subjects who reported bleeding gums, gums’ irritation, rough tooth surface and gum disease decreased steadily with age, there is an evident linear trend with high statistical significant difference among various age groups (P < 0.001). Furthermore, the proportions of subjects who answered correctly the periodontal knowledge questions related to plaque definition and its role in disease aetiology and prevention, decreased with age, with a high significant difference for all knowledge questions (P < 0.001). The prevalence of subjects with healthy periodontium (score 0) decreased with age, being 41.1% among 20–29‐year‐old group and 11.1% in 50–60‐year‐old group. Similar trend was seen for bleeding on probing (score 1) whilst calculus deposit (score 2) was the major problem in all age groups over 29 years. The prevalence of periodontal pockets was relatively low, with a range of 4.5–18.6% for shallow pockets (score 3) and 2.9–11.1% for deep pockets in younger and older age groups, respectively. Younger age group had more healthy sextants than older age groups. In addition, sextants with bleeding on probing were highest in 20–29‐year‐old age group (1.8) compared with that in 50–60‐year‐old age group (0.60) whilst those with calculus deposits varied slightly among different age groups (1.50–1.65). Sextants affected by shallow or deep pockets increased steadily with age. Such a finding was also noted in excluded sextants, with an increase from 0.15 at 20–29 year‐old group to 0.94 at the age of 50–60 years. It is concluded that health knowledge and status related to periodontal disease is still poor in northern Jordan. Therefore, dental health education provision is necessary to improve oral health knowledge and conditions among population in general and among older population in particular.
Clinical comparison of an adhesive precoated vs. an uncoated ceramic bracket system
Authors: Verstrynge A. , Willems G. , Ghesquiere A.
Published: November 2004

Structured Abstract

Authors – Verstrynge A, Ghesquiere A, Willems G.

Objectives – To clinically evaluate the bond characteristics of the adhesive precoated ceramic (APC) Clarity bracket and compare it with the uncoated Clarity bracket used with Transbond XT bonding system.

Design – A randomized clinical trial.

Settings and Sample Population – Department of Orthodontics, Katholieke Universiteit Leuven. Twenty patients requiring bonded fixed orthodontic appliance from central incisor to second bicuspid.

Experimental Variable – APC Clarity brackets were bonded in the first and fourth quadrant of group 1 and in the second and third quadrant of group 2. Uncoated Clarity brackets were bonded with Transbond XT in the second and third quadrant of group 1 and in the first and fourth quadrant of group 2.

Outcome Measure – During orthodontic treatment, bracket failures were registered. At debonding, the tooth surfaces were intraorally scored according to the Adhesive Remnant Index.

Results – During treatment one tie wing fractured in the APC group and no bond failures occurred. At debonding, no significant differences between the APC and Transbond XT could be shown in any of the quadrants.

Conclusion – The two groups performed identically.
Self‐reported denture hygiene of a sample of edentulous attendees at a University dental clinic and the relationship to the condition of the oral tissues
Authors: Marchini L. , Tamashiro E. , Nascimento D. F. F. , Cunha V. P. P.
Published: November 2004

Objectives:  This article aims to evaluate the habits of hygiene and usage of a sample group of Brazilian wearers of complete dentures.

Design:  This study has a cross‐sectional design.

Setting:  It was performed in the dental clinic of the University of Mogi das Cruzes.

Subjects:  A sample group of 236 complete denture wearers was selected.

Intervention:  The individuals were interviewed and clinically examined.

Main outcome measures:  Most commonly used hygiene habits were assessed and recorded. Possible relationships between habits and oral conditions were also assessed.

Results:  Dental treatment was sought by 43.6% 10 years after their last dental appointment, 77.5% declared they had been given no instructions regarding the hygiene of their dentures, only 22.9% said they had been instructed about oral hygiene, and 91.9% stated they had not been told to return for periodical review visits. Denture stomatitis was found in 42.4%, although 89% of these patients presented with no symptoms; 98.7% of the group brushed their dentures; 27.1% regularly immersed their dentures in chemical products; 26.3% removed their dentures overnight. A positive relationship was observed between the lack of recommendations regarding oral and denture cleansing and the presence of denture‐related stomatitis and hyperplasia. Family income and periodicity of visits to the dentist were also found to be related.

Conclusion:  Mechanical cleaning is the most prevalent method of hygiene. Self‐reported lack of guidance to oral care was statistically related to inflammatory oral conditions. Future research is necessary to clarify possible causal role between these factors.
Oral health promotion programme for older migrant adults
Authors: Mariño Rodrigo , Calache Hanny , Wright Clive , Schofield Margot , Minichiello Victor
Published: November 2004

Objectives:  This study evaluated the impact of a community‐based oral health promotion programme on the use of oral health services, oral health knowledge, attitudes, and practices of older Greek and Italian adults attending community clubs and living in Melbourne, Australia.

Methods:  The oral health promotion intervention consisted of three components: a series of oral health seminars, held at the clubs in the participants’ native languages, the provision of oral health care products, and the production of oral health information sheets. The intervention programme was known as the Oral Health Information Seminars/Sheets (ORHIS). The content of each session was determined following suggestions and findings from the data collected.

A pre‐test–post‐test non‐equivalent control group quasi‐experimental design was chosen to evaluate the intervention. A total of 520 independent‐living older adults, members of Greek or Italian social clubs participated in this evaluation.

Results:  Participants who took part in the intervention responded with higher levels of achievement than those in the control groups. After controlling for baseline variables, experimental groups were significantly more likely than the control groups at post‐test to have improved oral health attitudes, oral health knowledge, and self‐assessed physical health status, as well as, self‐reported oral hygiene practices and use of oral health services.

Discussion:  The ORHIS approach was successful within the setting of social clubs, and highly acceptable to these communities. As such, it represents a helpful approach for the design of (oral) health interventions in older adults. Further research is required to test the long‐tem impact including the economic evaluation of the ORHIS approach.
Endodontic considerations in the elderly
Authors: Allen P. Finbarr , Whitworth John M.
Published: November 2004

Tooth retention has increased significantly in older adults, and dentists are now challenged by the need to preserve critical teeth. There will be a need to consider endodontic therapy, and this paper describes how successful endodontics can be provided for elderly patients. Strategic treatment planning is essential, and preservation of key teeth will facilitate satisfactory oral function for elderly patients. These teeth may be important in achieving and maintaining an intact anterior dental arch, for removable partial denture retention or preservation of alveolar bone. In some cases, this can only be achieved if endodontic procedures are undertaken. When infection of a root canal is present, there is no reason why good quality endodontic therapy should not work in a healthy elderly patient. Elimination of infection can be challenging in narrow root canals, and a systematic approach for improving access into and negotiating these canals is outlined.
Radiological imaging to improve the emergency department diagnosis of acute appendicitis
Authors: Rosengren David , Brown Anthony FT , Chu Kevin
Published: November 2004

Objectives:  To determine the institution's current non‐therapeutic (negative) appendicectomy rate; the frequency of clinical predictors for appendicitis in patients who underwent appendicectomy; and the utilization and accuracy of ultrasound scans (USS) and computed tomography (CT) in the diagnosis of appendicitis.

Methods:  A retrospective chart review was conducted in an adult, metropolitan teaching hospital. Patients who presented to the ED and underwent an appendicectomy over a 12‐month period were analysed. Symptoms and signs predictive of appendicitis, results of USS and CT scans if performed, and histopathology findings were abstracted from patient records.

Results:  Two hundred and forty patients had appendicectomies, 147 (61%) were male and the median age was 25 years (range 14–78 years). The negative appendicectomy rate was 14.3% (95% CI 9.1–21.0%) and 18.3% (95% CI 11.0–26.7%) in males and females, respectively. Abdominal pain shifting to the right iliac fossa (RIF), anorexia and RIF rebound tenderness were found more frequently in patients with positive than negative appendicectomies (P < 0.05). USS and CT scans were performed in 68 (28%) and 15 (9.5%) patients, respectively. The likelihood ratio for appendicitis in patients with a normal USS or a normal CT scan was 0.83 (95% CI 0.56–1.24) and 0.08 (95% CI 0.01–0.60), respectively. There were no false positive CT scan results.

Conclusion:  Computed tomoraphy scanning should play an increasing role in the ED management of suspected appendicitis. Our negative appendicectomy rate could potentially be halved by the introduction of CT scans in the diagnostic work up of these patients.
Epistaxis
Authors: M Middleton Paul
Published: November 2004

Epistaxis is a frequent complaint, and may often cause great anxiety in patients and clinicians. Epistaxis results from the interaction of factors that damage the nasal mucosal lining, affect the vessel walls, or alter the coagulability of the blood, and which may be categorized into environmental, local, systemic and medication related. The knowledge of the first aid treatment of epistaxis is very poor, amongst not only the public, but also health professionals. Immediate emergency department management of epistaxis depends on prioritized assessment and treatment, including resuscitation if necessary, together with the application of relatively simple otolaryngological techniques. There is little high quality evidence regarding routine, alternative or adjunctive treatments.
Medical support for Adventure Racing
Authors: Greenland Keith
Published: November 2004

Adventure Racing has recently become a popular sport. In the past medical and paramedical staff were frequently positioned at checkpoints throughout the course of the race. Presently there is a developing role for appropriately qualified individuals to access the course by biking, running or swimming and then providing injured racers with on‐site first aid prior to their transfer to the nearest medical checkpoint. These medical staff should have not only appropriate first aid skills but also rock rescue techniques, rappelling and other basic rope work as well as be athletically fit. This article explores the new role and attempts to outline what characteristics may be required.
Bilateral theca lutein cysts: A rare cause of acute abdomen in pregnancy
Authors: Upadhyaya Geetika , Goswami Anu , Babu Suresh
Published: November 2004

Theca lutein cysts (hyperreactio luteinalis) are benign cysts usually associated with molar pregnancy. We report a case of bilateral theca lutein cysts with normal intrauterine singleton pregnancy presenting as an acute abdomen requiring surgical intervention. Laparotomy revealed bilateral theca lutein cysts one of which was torted, necessitating salpingo‐ovariotomy.
Inhibition of the Protease Activity of the Light Chain of Type A Botulinum Neurotoxin by Aqueous Extract from Stinging Nettle (Urtica dioica) Leaf
Authors: Gul Nizamettin , Ahmed S. Ashraf , Smith Leonard A.
Published: November 2004

Abstract: We investigated the inhibitory effect of stinging nettle leaf extract on the protease activity of botulinum neurotoxin type A and B light chains. The nettle leaf infusion was fractionated and HPLC‐based enzymatic assays were performed to determine the capacity of each fraction to inhibit the protease activity of botulinum neurotoxin type A and B light chains. Assay results demonstrated that a water‐soluble fraction obtained from the nettle leaf infusion inhibited type A, but did not inhibit type B light chain protease activity. The inhibition mode of water soluble fraction against protease activity of type A light chain was analyzed and found to be a non‐competitive.
Effect of Hibiscus rosa sinensis Extract on Hyperproliferation and Oxidative Damage Caused by Benzoyl Peroxide and Ultraviolet Radiations in Mouse Skin
Authors: Sharma Sonia , Sultana Sarwat
Published: November 2004

Abstract: The present study was conducted to investigate the ameliorative potential of Hibiscus rosa sinensis extract in mice skin. Combination of a single topical application of benzoyl peroxide (20 mg/0.2 ml/animal) followed by ultraviolet radiations (0.420 J/m2/s) was used to induce hyperproliferation and oxidative stress. Single benzoyl peroxide application prior to ultraviolet B radiations exposure caused significant depletion in the detoxification and antioxidant enzymes, while malondialdehyde formation, hydrogen peroxide content, ornithine decarboxylase activity and DNA synthesis were raised significantly. However, pretreatment of H. rosa sinensis extract (3.5 mg and 7 mg/ kg b.wt.) partly restored the levels of cellular protective enzymes (P<0.05). Besides, malondialdehyde formation and hydrogen peroxide content (P<0.05) were statistically significantly reduced at both doses. The ornithine decarboxylase activity and thymidine incorporation in DNA were also reduced dose dependently (P<0.05) by the plant extract. Therefore, we propose that H. rosa sinensis extract exerts a protective effect against the tumour promotion stage of cancer development.
Cooperative effect of S4–S5 loops in domains D3 and D4 on fast inactivation of the Na+ channel
Authors: Oana Popa M. , Alekov Alexi K. , Bail Sigrid , Lehmann‐Horn Frank , Lerche Holger
Published: November 2004

Cytoplasmic S4–S5 loops have been shown to be involved in fast inactivation of voltage‐gated ion channels. We studied mutations in these loops and their potential cooperative effects in domains D3 (N1151C, A1152C, I1160C/A) and D4 (F1473C, L1482C/A) of the human skeletal muscle Na+ channel α‐subunit (hNav1.4) using expression in tsA201 cells and the whole cell patch‐clamp technique. All cysteine mutations were accessible to intracellularly applied sulfhydryl reagents which considerably destabilized fast inactivation. For different combinations of corresponding D3/D4 double mutations, fast inactivation could be almost completely removed. Thermodynamic cycle analysis indicated an additive effect for N1151C/F1473C and a significant cooperative effect for I1160/L1482 double mutations. Application of oxidizing reagents such as Cu‐phenanthroline to link two cysteines via a disulfide bridge did not reveal evidence for a direct physical interaction of cysteines in D3 and D4. In addition to the pronounced alterations of fast inactivation, mutations of I1160 shifted steady‐state activation in the hyperpolarizing direction and slowed the kinetics of both activation and deactivation. Sulfhydryl reagents had charge‐dependent effects on I1160C suggesting interaction with negative charges in another protein region. We conclude that fast inactivation of the Na+ channel involves both S4–S5 loops in D3 and D4 in a cooperative manner. D3/S4–S5 also plays an important role in activation and deactivation.
Developmental programming of the metabolic syndrome by maternal nutritional imbalance: how strong is the evidence from experimental models in mammals?
Authors: Armitage James A. , Khan Imran Y. , Taylor Paul D. , Nathanielsz Peter W. , Poston Lucilla
Published: November 2004

The incidence of the metabolic syndrome, a cluster of abnormalities focusing on insulin resistance and associated with high risk for cardiovascular disease and diabetes, is reaching epidemic proportions. Prevalent in both developed and developing countries, the metabolic syndrome has largely been attributed to altered dietary and lifestyle factors that favour the development of central obesity. However, population‐based studies have suggested that predisposition to the metabolic syndrome may be acquired very early in development through inappropriate fetal or neonatal nutrition. Further evidence for developmental programming of the metabolic syndrome has now been suggested by animal studies in which the fetal environment has been manipulated through altered maternal dietary intake or modification of uterine artery blood flow. This review examines these studies and assesses whether the metabolic syndrome can be reliably induced by the interventions made. The validity of the different species, diets, feeding regimes and end‐point measures used is also discussed.
Infusion of collaborative inquiry throughout a biology curriculum increases student learning: a four-year study of “Teams and Streams”
Authors: Luckie Douglas B. , Maleszewski Joseph J. , Loznak Sarah D. , Krha Marija
Published: November 2004

Are traditional laboratories in the core introductory biology courses teaching physiology majors the art and trade of science, or simply leaving them with a memory of trivial experiments done for unknown reasons? Our students, a population dominated by premed and physiology majors, think the latter and have encouraged us to challenge this model, and it turns out scientists and education researchers agree with our students ( 4 , 31 , 32 ). In an effort to remedy this, we began a long-term redesign of the introductory biology sequence to become what is now a sequence of inquiry laboratories we term “Teams and Streams” (TS). In these TS inquiry labs, student research teams pose a scientific question/hypothesis, propose an experimental design, perform multi-week investigations and then present their findings in various forms (web, interviews, and papers). The response to this classroom laboratory design has been overwhelmingly positive. In a qualitative study of student opinion (where 260 student responses were studied), surveys conducted at the end of semesters where traditional scripted labs were used ( n = 70 comments) had predominately negative opinions (80% negative responses), whereas the reverse was true for students ( n = 190 comments) who participated in courses using the TS inquiry labs (78% positive responses). In a quantitative assessment of content knowledge, students who participated in new TS inquiry labs ( n = 245) outscored their peers in traditional labs ( n = 86) on Medical College Admission Test-style standardized exams (59.3 ± 0.8% vs. 48.9 ± 1.3%, respectively; P < 0.0001). We believe these quantitative data support the qualitative findings and suggest the TS inquiry lab approach increases student learning.
Generation of resting membrane potential
Authors: Wright Stephen H.
Published: November 2004

This brief review is intended to serve as a refresher on the ideas associated with teaching students the physiological basis of the resting membrane potential. The presentation is targeted toward first-year medical students, first-year graduate students, or senior undergraduates. The emphasis is on general concepts associated with generation of the electrical potential difference that exists across the plasma membrane of every animal cell. The intention is to provide students a general view of the quantitative relationship that exists between 1) transmembrane gradients for K+ and Na+ and 2) the relative channel-mediated permeability of the membrane to these ions.
Regulation of intracellular pH
Authors: Boron Walter F.
Published: November 2004

The approach that most animal cells employ to regulate intracellular pH (pHi) is not too different conceptually from the way a sophisticated system might regulate the temperature of a house. Just as the heat capacity (C) of a house minimizes sudden temperature (T) shifts caused by acute cold and heat loads, the buffering power (β) of a cell minimizes sudden pHishifts caused by acute acid and alkali loads. However, increasing C (or β) only minimizes T (or pHi) changes; it does not eliminate the changes, return T (or pHi) to normal, or shift steady-state T (or pHi). Whereas a house may have a furnace to raise T, a cell generally has more than one acid-extruding transporter (which exports acid and/or imports alkali) to raise pHi. Whereas an air conditioner lowers T, a cell generally has more than one acid-loading transporter to lower pHi. Just as a house might respond to graded decreases (or increases) in T by producing graded increases in heat (or cold) output, cells respond to graded decreases (or increases) in pHiwith graded increases (or decreases) in acid-extrusion (or acid-loading) rate. Steady-state T (or pHi) can change only in response to a change in chronic cold (or acid) loading or chronic heat (or alkali) loading as produced, for example, by a change in environmental T (or pH) or a change in the kinetics of the furnace (or acid extrudes) or air conditioner (or acid loaders). Finally, just as a temperature-control system might benefit from environmental sensors that provide clues about cold and heat loading, at least some cells seem to have extracellular CO2or extracellular HCO3−sensors that modulate acid-base transport.
In the beginning, there was the cell: cellular homeostasis
Authors: Romero Michael F.
Published: November 2004

In the past 5 years, the biomedical, scientific community has sequenced the genomes of several organisms (including Homo sapiens), has cloned entire organisms and has determine the molecular structures for several membrane proteins. These advances combined with the advances in technology enabling high-throughput drug screening, gene expression readout using DNA chips and evolving proteomic techniques, make it imperative that physiologist and biomedical professionals understand the basis of cellular function and homeostasis. The Cellular Homeostasis Refresher Course at Experimental Biology 2004 in Washington, DC, was designed to fulfill this need. The specific topics covered were 1) generation of membrane potential, 2) an update on cellular mechanisms of ion homeostasis, channels and transporters, and 3) cellular volume homeostasis, and regulation of intracellular pH.
Conservation genomics: applying whole genome studies to species conservation efforts
Authors: Ryder O.A.
Published: November 2004

Studies of complete genomes are leading to a new understanding of the biology of mammals and providing ongoing insights into the fundamental aspects of the organization and evolution of biological systems. Comparison of primate genomes can identify aspects of their organization, regulation and function that appeared during the primate radiation, but without comparison to more evolutionarily distant mammals and other vertebrates, highly conserved aspects of genome architecture will not be accurately identified nor will the lineage-specific changes be identified as such. Many species of primates face risks of extinction; yet the knowledge of their genomes will provide a deeper understanding of primate adaptations, human origins, and provide the framework for discoveries anticipated to improve human medicine. The great apes, the closest relatives of the human species, are among the most vulnerable and most important for human medical studies. However, apes are not the only species whose genomic information will enrich humankind. Comparative genomic studies of endangered species can benefit conservation efforts on their behalf. Increased knowledge of genome makeup and variation in endangered species finds conservation application in population evaluation monitoring and management, understanding phylozoogeography, can enhance wildlife health management, identify risk factors for genetic disorders, and provide insights into demographic management of small populations in the wild and in captivity.
The primates of the Neotropics: genomes and chromosomes
Authors: Seuánez H.N. , Bonvicino C.R. , Moreira M.A.M.
Published: November 2004

The classification of neotropical primates has been controversial. Different arrangements have been proposed, depending on taxonomic criteria and on the traits selected for phylogenetic reconstructions. These include gross morphologic characters, karyotypic attributes and DNA sequence data of nuclear and mitochondrial genes and of repetitive genomic components. These approaches have substantially clarified the main intergeneric relationships although several intrageneric arrangements still remain to be elucidated. In this review, we compare karyologic and molecular data of this speciose group.
Identity by descent and DNA sequence variation of human SINE and LINE elements
Authors: Salem A.-H. , Ray D.A. , Batzer M.A.
Published: November 2004

To test the hypothesis that Alu and L1 elements are genetic characters that are essentially homoplasy-free, we sequenced a total of five human L1 elements and eleven recently integrated Alu elements from 160 chromosomes (80 individuals representing four diverse human populations). Analysis of worldwide samples at L1 loci revealed 292 segregating sites and a nucleotide diversity of 0.0050. For Ya5 Alu loci, there were 129 segregating sites and nucleotide diversity was estimated at 0.0045. The Alu and L1 sequence diversity varied element to element. No completely or partially deleted Alu or L1 alleles were identified during the analysis. These data suggest that mobile element insertions are identical by descent characters for the study of human population genetics.
Nucleotide sequence comparison of a chromosome rearrangement on human chromosome 12 and the corresponding ape chromosomes
Authors: Shimada M.K. , Kim C.-G. , Kitano T. , Ferrell R.E. , Kohara Y. , Saitou N.
Published: November 2004

Chromosome rearrangement has been considered to be important in the evolutionary process. Here, we demonstrate the evolutionary relationship of the rearranged human chromosome 12 and the corresponding chromosome XII in apes (chimpanzee, bonobo, gorilla, orangutan, and gibbon) by examining PCR products derived from the breakpoints of inversions and by conducting shotgun sequencing of a gorilla fosmid clone containing the breakpoint and a “duplicated segment” (duplicon). We confirmed that a pair of 23-kb duplicons flank the breakpoints of inversions on the long and short arms of chimpanzee chromosome XII. Although only the 23-kb duplicon on the long arm of chimpanzee chromosome XII and its telomeric flanking sequence are found to be conserved among the hominoids (human, great apes, and gibbons), the duplicon on the short arm of chimpanzee chromosome XII is suggested to be the result of a duplication from that on the long arm. Furthermore, the shotgun sequencing of a gorilla fosmid indicated that the breakpoint on the long arm of the gorilla is located at a different position 1.9 kb from that of chimpanzee. The region is flanked by a sequence homologous to that of human chromosome 6q22. Our findings and sequence analysis suggest a close relationship between segmental duplication and chromosome rearrangement (or breakpoint of inversion) in Hominoidea. The role of the chromosome rearrangement in speciation is also discussed based on our new results.
Genomic structure and paralogous regions of the inversion breakpoint occurring between human chromosome 3p12.3 and orangutan chromosome 2
Authors: Yue Y. , Grossmann B. , Tsend-Ayush E. , Grützner F. , Ferguson-Smith M.A. , Yang F. , Haaf T.
Published: November 2004

Intrachromosomal duplications play a significant role in human genome pathology and evolution. To better understand the molecular basis of evolutionary chromosome rearrangements, we performed molecular cytogenetic and sequence analyses of the breakpoint region that distinguishes human chromosome 3p12.3 and orangutan chromosome 2. FISH with region-specific BAC clones demonstrated that the breakpoint-flanking sequences are duplicated intrachromosomally on orangutan 2 and human 3q21 as well as at many pericentromeric and subtelomeric sites throughout the genomes. Breakage and rearrangement of the human 3p12.3-homologous region in the orangutan lineage were associated with a partial loss of duplicated sequences in the breakpoint region. Consistent with our FISH mapping results, computational analysis of the human chromosome 3 genomic sequence revealed three 3p12.3-paralogous sequence blocks on human chromosome 3q21 and smaller blocks on the short arm end 3p26→p25. This is consistent with the view that sequences from an ancestral site at 3q21 were duplicated at 3p12.3 in a common ancestor of orangutan and humans. Our results show that evolutionary chromosome rearrangements are associated with microduplications and microdeletions, contributing to the DNA differences between closely related species.
Cytochrome b polymorphisms and population structure of two species of Alouatta (Primates)
Authors: Nascimento F.F. , Bonvicino C.R. , da Silva F.C.D. , Schneider M.P.C. , Seuánez H.N.
Published: November 2004

We carried out a phylogenetic and population study in Alouatta caraya and Alouatta belzebul based on cytochrome b DNA sequence data. Maximum Parsimony and Median-Joining analyses grouped A. caraya from different localities showing a population structure in accordance with geographic distribution. The relation between A. caraya haplotypes could be explained with respect to the species range in the Cerrado, one of the most ancient morphoclimatic domains of South America, and the Chaco. Conversely, A. belzebul from the Amazonas and Atlantic forests grouped in a paraphyletic arrangement without an evident geographic pattern. Recent geologic events resulting in the separation of A. belzebul might explain why these geographically distant groups shared similar haplotypes and why ancestral polymorphisms might have been maintained in this species. Time of divergence estimates indicated that the splitting of the Alouatta lineage leading to A. caraya occurred some 4.58 MYA while the lineage leading to A. belzebul emerged 4.14 MYA.
The impact of chromosome sorting and painting on the comparative analysis of primate genomes
Authors: Ferguson-Smith M.A. , Yang F. , Rens W. , O’Brien P.C.M.
Published: November 2004

Chromosome sorting by flow cytometry is the main source of chromosome-specific DNA for the production of painting probes. These probes have been used for cross-species in situ hybridization in the construction of comparative maps, in the study of karyotype evolution and phylogenetics, in delineating territories in interphase nuclei, and in the analysis of chromosome breakpoints. We review here the contributions that this technology has made to the analysis of primate genomes.
Investigation of marmoset hybrids (Cebuella pygmaea × Callithrix jacchus) and related Callitrichinae (Platyrrhini) by cross-species chromosome painting and comparative genomic hybridization
Authors: Neusser M. , Münch M. , Anzenberger G. , Müller S.
Published: November 2004

We report on the cytogenetics of twin offspring from an interspecies cross in marmosets (Callitrichinae, Platyrrhini), resulting from a pairing between a female Common marmoset (Callithrix jacchus, 2n = 46) and a male Pygmy marmoset (Cebuella pygmaea, 2n = 44). We analyzed their karyotypes by multi-directional chromosome painting employing human, Saguinus oedipus and Lagothrix lagothricha chromosome-specific probes. Both hybrid individuals had a karyotype with a diploid chromosome number of 2n = 45. As a complementary tool, interspecies comparative genomic hybridization (iCGH) was performed in order to screen for genomic imbalances between the hybrids and their parental species, and between Callithrix argentata and S. oedipus, respectively. These genomic imbalances were confined to centromeric and telomeric heterochromatin, while euchromatic chromosome regions appeared balanced in all species investigated. When comparing marmosets and tamarins, sequence divergence of centromeric heterochromatin was already clearly noticeable. In the C. argentata and C. pygmaea genomes numerous subtelomeric regions were affected by amplification of different repetitive sequences. Cross-species FISH with a microdissection-derived C. pygmaea repetitive probe revealed species specificity of this repetitive sequence at the molecular cytogenetic level of resolution.
Application of molecular cytogenetics for chromosomal evolution of the Lemuriformes (Prosimians)
Authors: Warter S. , Hauwy M. , Dutrillaux B. , Rumpler Y.
Published: November 2004

R-banding chromosomal studies of 21 species of Lemuriformes allowed us to reconstruct the presumed ancestral karyotype of all the Lemuriformes except for Daubentoniidae and permitted the construction of their phylogenetic tree. Chromosome painting with fluorescently labeled heterologous DNA probes permitted comparative chromosome maps to be established. The Zoo-FISH method was used to reassess the karyotypes of 22 species or subspecies. While our results largely confirm the previous reconstruction of the ancestral karyotype, they resulted in a modification of the previously established phylogenetic tree. The Daubentoniidae emerged first followed by the divergence of the families Cheirogaleidae, Indriidae, Lepilemuridae and Lemuridae. Eight chromosome rearrangements occurred in all Lemuriformes except for Daubentoniidae in the common trunk. The present findings do not allow us to propose the occurrence of any rearrangement common to Daubentoniidae and other Lemuriformes, and probably other Prosimii. Conserved syntenies previously described in various mammalian orders were also conserved, while others were specific to the Lemuriformes.
Comparative mapping of human claudin-1 (CLDN1) in great apes
Authors: Nanda I. , Krämer F. , Weber B.H.F. , Schempp W. , Schmid M.
Published: November 2004

The gene encoding claudin-1 (CLDN1) has been mapped to human chromosome 3 (HSA3; 3q28→q29) using a radiation hybrid panel. Employing fluorescence in situ hybridization (FISH) we here show that a human P1-derived artificial chromosome (PAC) containing CLDN1 detects the orthologous sites in chromosomes of the great apes, chimpanzee, gorilla, and orangutan. Furthermore, the chromosomal position of CLDN1 was determined in mouse chromosomes by FISH. The position of fluorescent signals is confined to a single chromosomal site in both great apes and mouse and in each case maps to the chromosomal region that has conserved synteny with HSA3 (PTR2q28, GGO2q28, PPY2q38 and MMU16B1). Using a gene-specific probe our results are consistent with reports of the striking similarity of great ape and human genomes as illustrated previously by chromosome painting.
Chimpanzee chromosomes: retrotransposable compound repeat DNA organization (RCRO) and its influence on meiotic prophase and crossing-over
Authors: Hirai H. , Matsubayashi K. , Kumazaki K. , Kato A. , Maeda N. , Kim H.-S.
Published: November 2004

The terminal C-bands that are a specific feature of chimpanzee chromosomes were dissected using a molecular cytogenetic technique, PRINS, with primers for telomeric sequences, subterminal satellite, and retrotransposable elements (HERV-K and -W). These DNA elements jointly formed a large block of retrotransposable compound repeat DNA organization (RCRO) at the terminal C-band regions of 30 chromosomes, and are also located at the centromeric regions of some chromosomes. Additionally, a block consisting of all members of the RCRO has transposed to the middle (q31.1) of the long arm of chromosome 6, and three members, the subterminal satellite and the two HERVs, have integrated into the proximal region (q14.4) of the long arm of chromosome 14. Terminal RCROs seem to induce and prolong the bouquet stage in meiotic prophase, and to affect chiasma formation, together with interstitial RCROs. It is also postulated that RCROs may cause a position effect to gene expression, resulting in gene silencing and/or late replication.
Fine structure and meiotic behaviour of the male multiple sex chromosomes in the genus Alouatta
Authors: Solari A.J. , Rahn M.I.
Published: November 2004

The meiotic cytology and fine structure of the sex multiples in males from two species of the genus Alouatta are presented and compared with descriptions from other species of this genus. As shown in pachytene by synaptonemal complex analysis and in metaphase I by spreading, there is a quadrivalent in male meiosis in A. caraya, which is formed by an X1X2Y1Y2 complex, while in A. palliata there is a trivalent formed by an X1X2Y1 complex. Chromosome painting with human probes shows that A. caraya sex multiples share the same components as those of A. seniculus sara and A. seniculus arctoidea. However, as shown here for A. palliata and by others in A. fusca, there are differences among the multiples of some species. It is shown that in this genus there are several varieties of sex multiples that share some features, and that the origin of these multiples is most probably a primitive development in the genus Alouatta.
South China Sea Wind-Wave Characteristics. Part I: Validation of Wavewatch-III Using TOPEX/Poseidon Data
Authors: Chu Peter C. , Qi Yiquan , Chen Yuchun , Shi Ping , Mao Qingwen
Published: November 2004

A full-spectral third-generation ocean wind-wave model, Wavewatch-III, has been implemented in the South China Sea (SCS) for investigating wind-wave characteristics. This model was developed at the Ocean Modeling Branch of the National Centers for Environmental Prediction (NCEP). The NASA QuickSCAT data (0.25° resolution) 2 times daily were used to simulate the wind waves for the entire year of 2000. The significant wave heights from Wavewatch-III are compared to the TOPEX/Poseidon (T/P) significant wave height data over the satellite crossover points in SCS. The model errors of significant wave height have Gaussian-type distribution with a small mean value of 0.02 m (almost no bias). The model errors are comparable to the T/P altimeter accuracy (0.5 m) in the central SCS and are smaller than the T/P altimeter accuracy in the northern and southern SCS, which indicates the capability of Wavewatch-III for SCS wave simulation.
DOV 216,303, a “Triple” Reuptake Inhibitor: Safety, Tolerability, and Pharmacokinetic Profile
Authors: Beer Bernard , Stark Jill , Krieter Philip , Czobor Pal , Beer Gary , Lippa Arnold , Skolnick Phil
Published: November 2004

This report describes the first evaluation in humans of DOV 216,303, a putative antidepressive that inhibits the reuptake of norepinephrine, serotonin, and dopamine. Subjects received single oral doses of 5 to 150 mg of DOV 216,303 or placebo. At 150 mg, 4 of 7 subjects reported gastrointestinal disturbances. In the multiple‐dose phase of the evaluation, subjects received total daily doses of 50, 75, or 100 mg of DOV 216,303 or placebo for 10 days. At a total daily dose of 100 mg, gastrointestinal disturbances were reported in 4 of 6 volunteers. In both the single‐ and multiple‐dose evaluations, no significant changes were noted in vital signs, electrocardiogram, hematology, or clinical chemistry. DOV 216,303 was rapidly absorbed (plasma tmax of 0.7–1.2 hours and t1/2 of 3.3–4.4 hours), with dose‐proportional Cmax and AUC values. Furthermore, no remarkable difference was apparent in either the Cmax or AUC of DOV 216,303 following 1 and 10 days of dosing. The present results demonstrate that DOV 216,303 is safe and well tolerated both at single doses of up to 100 mg and multiple doses of up to 100 mg/day for 10 days. Plasma concentrations of DOV 216,303 after doses >10 mg exceed its reported IC50 values for inhibition of biogenic amine reuptake.
Utility of Sparse Concentration Sampling for Citalopram in Elderly Clinical Trial Subjects
Authors: Bies Robert R. , Feng Yan , Lotrich Francis E. , Kirshner Margaret A. , Roose Steven , Kupfer David J. , Pollock Bruce G.
Published: November 2004

The objective of this study was to evaluate whether the disposition of the selective serotonin reuptake inhibitor, citalopram, could be robustly captured using 1 to 2 concentration samples per subject in 106 patients participating in 2 clinical trials. Nonlinear mixed‐effects modeling was used to evaluate the pharmacokinetic parameters describing citalopram's disposition. Both a prior established 2‐compartment model and a de novo 1‐compartment pharmacokinetic model were used. Covariates assessed were concomitant medications, race, sex, age (22–93 years), and weight. Covariates affecting disposition were assessed separately and then combined in a stepwise manner. Pharmacokinetic characteristics of citalopram were well captured using this sparse sampling design. Two covariates (age and weight) had a significant effect on the clearance and volume of distribution in both the 1‐ and 2‐compartment pharmacokinetic models. Clearance decreased 0.23 L/h for every year of age and increased 0.14 L/h per kilogram body weight. It was concluded that hyper‐sparse sampling designs are adequate to support population pharmacokinetic analysis in clinically treated populations. This is particularly valuable for populations such as the elderly, who are not typically available for pharmacokinetic studies.