bluuebunny/crossref_metadata_2025 huggingface dataset:
In vivo
T lymphocyte dynamics in humans and the impact of human T-lymphotropic virus 1 infection
Authors: Asquith Becca , Zhang Yan , Mosley Angelina J. , de Lara Catherine M. , Wallace Diana L. , Worth Andrew , Kaftantzi Lambrini , Meekings Kiran , Griffin George E. , Tanaka Yuetsu , Tough David F. , Beverley Peter C. , Taylor Graham P. , Macallan Derek C. , Bangham Charles R. M.
Published: May 2007

Human T-lymphotropic virus type 1 (HTLV-1) is a persistent CD4
+
T-lymphotropic retrovirus. Most HTLV-1-infected individuals remain asymptomatic, but a proportion develop adult T cell leukemia or inflammatory disease. It is not fully understood how HTLV-1 persists despite a strong immune response or what determines the risk of HTLV-1-associated diseases. Until recently, it has been difficult to quantify lymphocyte kinetics in humans
in vivo
. Here, we used deuterated glucose labeling to quantify
in vivo
lymphocyte dynamics in HTLV-1-infected individuals. We then used these results to address four questions. (
i
) What is the impact of HTLV-1 infection on lymphocyte dynamics? (
ii
) How does HTLV-1 persist? (
iii
) What is the extent of HTLV-1 expression
in vivo
? (
iv
) What features of lymphocyte kinetics are associated with HTLV-1-associated myelopathy/tropical spastic paraparesis? We found that CD4
+
CD45RO
+
and CD8
+
CD45RO
+
T lymphocyte proliferation was elevated in HTLV-1-infected subjects compared with controls, with an extra 10
12
lymphocytes produced per year in an HTLV-1-infected subject. The
in vivo
proliferation rate of CD4
+
CD45RO
+
cells also correlated with
ex vivo
viral expression. Finally, the inflammatory disease HTLV-1-associated myelopathy/tropical spastic paraparesis was associated with significantly increased CD4
+
CD45RO
+
cell proliferation. We suggest that there is persistent viral gene expression
in vivo
, which is necessary for the maintenance of the proviral load and determines HTLV-1-associated myelopathy/tropical spastic paraparesis risk.
The G protein-coupled receptor regulatory kinase GPRK2 participates in Hedgehog signaling in
Drosophila
Authors: Molnar Cristina , Holguin Helena , Mayor Federico , Ruiz-Gomez Ana , de Celis Jose F.
Published: May 2007

Signaling by Smoothened (Smo) plays fundamental roles during animal development and is deregulated in a variety of human cancers. Smo is a transmembrane protein with a heptahelical topology characteristic of G protein-coupled receptors. Despite such similarity, the mechanisms regulating Smo signaling are not fully understood. We show that Gprk2, a
Drosophila
member of the G protein-coupled receptor kinases, plays a key role in the Smo signal transduction pathway. Lowering
Gprk2
levels in the wing disc reduces the expression of Smo targets and causes a phenotype reminiscent of loss of
Smo
function. We found that Gprk2 function is required for transducing the Smo signal and that when
Gprk2
levels are lowered, Smo still accumulates at the cell membrane, but its activation is reduced. Interestingly, the expression of
Gprk2
in the wing disc is regulated in part by Smo, generating a positive feedback loop that maintains high Smo activity close to the anterior–posterior compartment boundary.
Mechanism and uses of a membrane peptide that targets tumors and other acidic tissues
in vivo
Authors: Andreev Oleg A. , Dupuy Allison D. , Segala Michael , Sandugu Srikanth , Serra David A. , Chichester Clinton O. , Engelman Donald M. , Reshetnyak Yana K.
Published: May 2007

The pH-selective insertion and folding of a membrane peptide, pHLIP [pH (low) insertion peptide], can be used to target acidic tissue
in vivo
, including acidic foci in tumors, kidneys, and inflammatory sites. In a mouse breast adenocarcinoma model, fluorescently labeled pHLIP finds solid acidic tumors with high accuracy and accumulates in them even at a very early stage of tumor development. The fluorescence signal is stable for >4 days and is approximately five times higher in tumors than in healthy counterpart tissue. In a rat antigen-induced arthritis model, pHLIP preferentially accumulates in inflammatory foci. pHLIP also maps the renal cortical interstitium; however, kidney accumulation can be reduced significantly by providing mice with bicarbonate-containing drinking water. The peptide has three states: soluble in water, bound to the surface of a membrane, and inserted across the membrane as an α-helix. At physiological pH, the equilibrium is toward water, which explains its low affinity for cells in healthy tissue; at acidic pH, titration of Asp residues shifts the equilibrium toward membrane insertion and tissue accumulation. The replacement of two key Asp residues located in the transmembrane part of pHLIP by Lys or Asn led to the loss of pH-sensitive insertion into membranes of liposomes, red blood cells, and cancer cells
in vivo
, as well as to the loss of specific accumulation in tumors. pHLIP nanotechnology introduces a new method of detecting, targeting, and possibly treating acidic diseased tissue by using the selective insertion and folding of membrane peptides.
Probing the folding intermediate of Rd‐apocyt b562 by protein engineering and infrared T‐jump
Authors: Wang Ting , Zhou Zheng , Bunagan Michelle R. , Du Deguo , Bai Yawen , Gai Feng
Published: May 2007

Small proteins often fold in an apparent two‐state manner with the absence of detectable early‐folding intermediates. Recently, using native‐state hydrogen exchange, intermediates that exist after the rate‐limiting transition state have been identified for several proteins. However, little is known about the folding kinetics from these post‐transition intermediates to their corresponding native states. Herein, we have used protein engineering and a laser‐induced temperature‐jump (T‐jump) technique to investigate this issue and have applied it to Rd‐apocyt b562, a four‐helix bundle protein. Previously, it has been shown that Rd‐apocyt b562 folds via an on‐pathway hidden intermediate, which has only the N‐terminal helix unfolded. In the present study, a double mutation (V16G/I17A) in the N‐terminal helix of Rd‐apocyt b562 was made to further increase the relative population of this intermediate state at high temperature by selectively destabilizing the native state. In the circular dichroism thermal melting experiment, this mutant showed apparent two‐state folding behavior. However, in the T‐jump experiment, two kinetic phases were observed. Therefore, these results are in agreement with the idea that a folding intermediate is populated on the folding pathway of Rd‐apocyt b562. Moreover, it was found that the exponential growth rate of the native state from this intermediate state is roughly (25 μsec)−1 at 65°C.
Conformational change of the methionine 20 loop of Escherichia coli dihydrofolate reductase modulates pKa of the bound dihydrofolate
Authors: Khavrutskii Ilja V. , Price Daniel J. , Lee Jinhyuk , Brooks Charles L.
Published: May 2007

We evaluate the pKa of dihydrofolate (H2F) at the N5 position in three ternary complexes with Escherichia coli dihydrofolate reductase (ecDHFR), namely ecDHFR(NADP+:H2F) in the closed form (1), and the Michaelis complexes ecDHFR(NADPH:H2F) in the closed (2) and occluded (3) forms, by performing free energy perturbation with molecular dynamics simulations (FEP/MD). Our simulations suggest that in the Michaelis complex the pKa is modulated by the Met20 loop fluctuations, providing the largest pKa shift in substates with a “tightly closed” loop conformation; in the “partially closed/open” substates, the pKa is similar to that in the occluded complex. Conducive to the protonation, tightly closing the Met20 loop enhances the interactions of the cofactor and the substrate with the Met20 side chain and aligns the nicotinamide ring of the cofactor coplanar with the pterin ring of the substrate. Overall, the present study favors the hypothesis that N5 is protonated directly from solution and provides further insights into the mechanism of the substrate protonation.
Structural determinants of nitroxide motion in spin‐labeled proteins: Tertiary contact and solvent‐inaccessible sites in helix G of T4 lysozyme
Authors: Guo Zhefeng , Cascio Duilio , Hideg Kálmán , Kálái Támás , Hubbell Wayne L.
Published: May 2007

A nitroxide side chain (R1) has been substituted at single sites along a helix–turn–helix motif in T4 lysozyme (residues 114–135). Together with previously published data, the new sites reported complete a continuous scan through the motif. Mutants with R1 at sites 115 and 118 were selected for crystallographic analysis to identify the structural origins of the corresponding two‐component EPR spectra. At 115, R1 is shown to occupy two rotamers in the room temperature crystal structure, one of which has not been previously reported. The two components in the EPR spectrum apparently arise from differential interactions of the two rotamers with the surrounding structure, the most important of which is a hydrophobic interaction of the nitroxide ring. Interestingly, the crystal structure at 100 K reveals a single rotamer, emphasizing the possibility of rotamer selection in low‐temperature crystal structures. Residue 118 is at a solvent‐inaccessible site in the protein core, and the structure of 118R1, the first reported for the R1 side chain at a buried site, reveals how the side chain is accommodated in an overpacked core.
Calcium‐induced folding of a fragment of calmodulin composed of EF‐hands 2 and 3
Authors: Lakowski Ted M. , Lee Gregory M. , Okon Mark , Reid Ronald E. , McIntosh Lawrence P.
Published: May 2007

Calmodulin (CaM) is an EF‐hand protein composed of two calcium (Ca2+)‐binding EF‐hand motifs in its N‐domain (EF‐1 and EF‐2) and two in its C‐domain (EF‐3 and EF‐4). In this study, we examined the structure, dynamics, and Ca2+‐binding properties of a fragment of CaM containing only EF‐2 and EF‐3 and the intervening linker sequence (CaM2/3). Based on NMR spectroscopic analyses, Ca2+‐free CaM2/3 is predominantly unfolded, but upon binding Ca2+, adopts a monomeric structure composed of two EF‐hand motifs bridged by a short antiparallel β‐sheet. Despite having an “even–odd” pairing of EF‐hands, the tertiary structure of CaM2/3 is similar to both the “odd–even” paired N‐ and C‐domains of Ca2+‐ligated CaM, with the conformationally flexible linker sequence adopting the role of an inter‐EF‐hand loop. However, unlike either CaM domain, CaM2/3 exhibits stepwise Ca2+ binding with a Kd1 = 30 ± 5 μM to EF‐3, and a Kd2 > 1000 μM to EF‐2. Binding of the first equivalent of Ca2+ induces the cooperative folding of CaM2/3. In the case of native CaM, stacking interactions between four conserved aromatic residues help to hold the first and fourth helices of each EF‐hand domain together, while the loop between EF‐hands covalently tethers the second and third helices. In contrast, these aromatic residues lie along the second and third helices of CaM2/3, and thus are positioned adjacent to the loop between its “even–odd” paired EF‐hands. This nonnative hydrophobic core packing may contribute to the weak Ca2+ affinity exhibited by EF‐2 in the context of CaM2/3.
Structure of the fungal β‐glucan‐binding immune receptor dectin‐1: Implications for function
Authors: Brown James , O'Callaghan Chris A. , Marshall Andrew S.J. , Gilbert Robert J.C. , Siebold Christian , Gordon Siamon , Brown Gordon D. , Jones E. Yvonne
Published: May 2007

The murine molecule dectin‐1 (known as the β‐glucan receptor in humans) is an immune cell surface receptor implicated in the immunological defense against fungal pathogens. Sequence analysis has indicated that the dectin‐1 extracellular domain is a C‐type lectin‐like domain, and functional studies have established that it binds fungal β‐glucans. We report several dectin‐1 crystal structures, including a high‐resolution structure and a 2.8 Å resolution structure in which a short soaked natural β‐glucan is trapped in the crystal lattice. In vitro characterization of dectin‐1 in the presence of its natural ligand indicates higher‐order complex formation between dectin‐1 and β‐glucans. These combined structural and biophysical data considerably extend the current knowledge of dectin‐1 structure and function, and suggest potential mechanisms of defense against fungal pathogens.
Induction of mictic females in the rotiferBrachionus: oocytes of amictic females respond individually to population‐density signal only during oogenesis shortly before oviposition
Authors: GILBERT JOHN J.
Published: May 2007

1. One at a time during the reproductive period of amictic females, oocytes fill with yolk and undergo a mitotic maturation division (oogenesis), are oviposited as single cells, and then develop parthenogenetically into females. Sexual reproduction inBrachionusand several other genera is initiated when amictic females are crowded and oviposit some eggs induced to differentiate into mictic females. Mictic females produce haploid eggs that can develop parthenogentically into males or be fertilised and develop into diapausing embryos called resting eggs.

2. This study examines the time when oocytes in amictic females respond to maternal population density. Is the fate of all oocytes in the germarium irreversibly determined during the early postnatal life of the mother, or is each oocyte labile until just before oviposition? In the former case, the probability of an amictic female producing a mictic daughter at any time throughout her reproductive period would reflect the population density she experienced while young and not that at the time she oviposited an egg.

3. Amictic females of two clones of a Florida strain ofB. calycifloruswere cultured singly from birth at a low or high density (in a large or small volume) until about halfway through their reproductive period and then switched (experimental treatment), or not (control treatment), to the other density condition. The results indicate that the female fate of an oocyte is determined by maternal population density during oogenesis. Eggs oviposited soon after transfer from low to high density had the same, or a higher, probability of becoming mictic females compared with those produced by control females kept at the high density; eggs oviposited after transfer from the high to the low density had the same low probability of becoming mictic females as those produced by control females kept at the low density.

4. Control females kept at the high density were less likely to produce mictic daughters as they aged. This decline is not because of a decreased propensity of older females to respond to crowding, as older females responded maximally when transferred from a low to a high population density.

5. As oocytes in amictic females respond to maternal population density only during oogenesis, there is a negligible lag between the population‐density signal in the environment and the commitment to sexual reproduction. This minimises the obligatory two‐generation lag between this signal and production of resting eggs, and thus reduces the possibility that crowding will lead to food limitation before production of these eggs.
Passive ultrasonic irrigation of the root canal: a review of the literature
Authors: Van Der Sluis L. W. M. , Versluis M. , Wu M. K. , Wesselink P. R.
Published: May 2007

Ultrasonic irrigation of the root canal can be performed with or without simultaneous ultrasonic instrumentation. When canal shaping is not undertaken the term passive ultrasonic irrigation (PUI) can be used to describe the technique. In this paper the relevant literature on PUI is reviewed from a MEDLINE database search.

Passive ultrasonic irrigation can be performed with a small file or smooth wire (size 10–20) oscillating freely in the root canal to induce powerful acoustic microstreaming. PUI can be an important supplement for cleaning the root canal system and, compared with traditional syringe irrigation, it removes more organic tissue, planktonic bacteria and dentine debris from the root canal. PUI is more efficient in cleaning canals than ultrasonic irrigation with simultaneous ultrasonic instrumentation. PUI can be effective in curved canals and a smooth wire can be as effective as a cutting K‐file. The taper and the diameter of the root canal were found to be important parameters in determining the efficacies of dentine debris removal. Irrigation with sodium hypochlorite is more effective than with water and ultrasonic irrigation is more effective than sonic irrigation in the removal of dentine debris from the root canal. The role of cavitation during PUI remains inconclusive. No detailed information is available on the influence of the irrigation time, the volume of the irrigant, the penetration depth of the instrument and the shape and material properties of the instrument. The influence of irrigation frequency and intensity on the streaming pattern as well as the complicated interaction of acoustic streaming with the adherent biofilm needs to be clarified to reveal the underlying physical mechanisms of PUI.
The (un)‐certainties of district nurses in the context of cultural diversity
Authors: Peckover Sue , Chidlaw Robert G.
Published: May 2007

Aim.  This paper is a report of part of a study exploring district nurses’ understanding and practices in relation to discrimination and inequalities issues.

Background.  Clients from minority ethnic groups continue to experience inequalities in health status and care provision. District nurses work with a wide range of clients, yet research has demonstrated inequities in service provision to clients from minority ethnic groups due to organizational, professional and personal constraints.

Methods.  A qualitative methodology was used to explore district nurses’ understanding of discrimination and equalities issues in the context of their work. Semi‐structured interviews were undertaken during 2003 with 18 district nurses employed in two primary care organizations in England providing healthcare services to a large and culturally diverse population.

Findings.  District nurses’ accounts were marked by uncertainty, suggesting that many lacked confidence when discussing issues of ethnicity and cultural difference. There were marked differences in the extent to which they appeared to recognize or engage with clients’ experiences of discrimination or inequalities, or to recognize inequitable service provision to clients from minority ethnic groups. Key issues were a lack of provision of district nursing services to some clients, and failures to meet clients’ language and communication needs, and although reflecting organizational constraints, such continuing inequities were largely unquestioned.

Conclusion.  There is a need for further educational and practice developments to enable district nurses to provide more equitable care to clients from culturally diverse communities, ensuring that the key professional discourse of individual care and advocacy are fully realized in their work with all clients.
A nurse‐ and pharmacist‐led treatment advice clinic for patients attending an HIV outpatient clinic
Authors: Griffiths C. , Miles K. , Aldam D. , Cornforth D. , Minton J. , Edwards S. , Williams I.
Published: May 2007

Aim.  This paper is a report of a study to map care pathways, examine the approach of different treatment advisors and explore the acceptability of a nurse‐ and pharmacist‐led treatment advice clinic in order to aid decision‐making for the future development and evaluation of the clinic.

Background.  High levels of adherence to antiretroviral drugs are a prerequisite for a successful and durable virological and immunological response to HIV. Treatment guidelines acknowledge that adherence is a process, not a single event, and that adherence support must be integrated into clinical follow‐up for all patients receiving these drugs.

Method.  Data were collected between September 2004 and January 2005 through 17 consultation observations and 10 patient interviews in a specialist treatment advice clinic located within a central London HIV outpatient clinic providing care for over 2200 patients, of whom more than 1300 are taking highly active antiretroviral therapy.

Findings.  The nurses and pharmacist had similar consultation approaches, although follow‐up care varied in extent. Benefits of the clinic approach included permitting patients to observe real tablets, tailoring regimens to lifestyles and telephone follow‐up. These factors, particularly telephone support, were perceived by patients to assist with adherence.

Conclusion.  The role of telephone support, perceived to assist with initial adherence, requires further investigation. Future work is also needed to explore the health economics of this approach and to determine the actual impact of the clinic on clinical and adherence outcomes.
Use of complementary and alternative medicine among people living with diabetes: literature review
Authors: Chang Hsiao‐yun , Wallis Marianne , Tiralongo Evelin
Published: May 2007

Aim.  This paper is a report of a literature review to explore the prevalence of complementary and alternative medicine use amongst people with diabetes to inform nursing practice, education and research.

Background.  Diabetes mellitus affects the entirety of a person's being and increasingly people use complementary and alternative medicine in conjunction with other medical treatments and lifestyle modifications to manage their condition and improve well‐being.

Methods.  The CINAHL, Medline, ProQuest nursing journals and Psych INFO databases were searched for the period 1990–2006 using identified keywords.

Results.  A total of 18 studies from nine countries were found. The results suggest that the prevalence of complementary and alternative medicine use among people with diabetes ranges from 17% to 72·8%. The most widely used therapies among diabetic populations are nutritional supplements, herbal medicines, nutritional advice, spiritual healing and relaxation techniques. The characteristics which influence complementary and alternative medicine use are age, duration of diabetes, degree of complications and self‐monitoring of blood glucose.

Conclusion.  Although inconsistency in the definition of complementary and alternative medicine and varying research designs make estimation of usage prevalence difficult, evidence suggests that a high proportion of people with diabetes use these therapies concurrently with conventional healthcare services. Healthcare professionals need to be aware of this issue and may need to incorporate complementary and alternative medicine information into patient assessment and intervention.
MHC class II Tetramer Guided Detection of Mycobacterium tuberculosis‐specific CD4+ T Cells in Peripheral Blood from Patients with Pulmonary Tuberculosis
Authors: Höhn H. , Kortsik C. , Zehbe I. , Hitzler W. E. , Kayser K. , Freitag K. , Neukirch C. , Andersen P. , Doherty T. M. , Maeurer M.
Published: May 2007

Novel diagnostic tools are needed to diagnose latent infection and to provide biologically meaningful surrogate markers to define cellular immune responses against Mycobacterium tuberculosis (MTB). Interferon gamma‐based assays have recently been developed in addition to the more than 100‐year‐old tuberculin skin test (TST) for the immune diagnosis of MTB in blood. The advent of soluble MHC/peptide tetramer molecules allows to objectively enumerate antigen‐specific T cells. We identified novel MHC class II‐restricted MTB epitopes and used HLA‐DR4 tetrameric complexes to visualize ex vivo CD4+ T cells directed against the antigens Ag85B and the 19‐kDa lipoprotein, shared between MTB and other Mycobacterium species, and CD4+ T cells which recognize the MTB‐associated ESAT‐6 antigen. MTB‐reactive CD4+ T cells reside predominantly in the CD45RA+ CD28+ and CD45− CD28+ T‐cell subset and recognize naturally processed and presented MTB epitopes. HLA‐DR4‐restricted, Ag85B or ESAT‐6‐specific CD4+ T cells show similar dynamics over time in peripheral blood mononuclear cells (PBMC) when compared with CD8+ T cells directed against the corresponding HLA‐A2‐presented MTB epitopes in patients with pulmonary MTB infection and subsequent successful therapy. This was not found to be true for T‐cell responses directed against the 19‐kDa lipoprotein. The dissection of the cellular immune response in M. tuberculosis infection will enable novel strategies for monitoring MTB vaccine candidates and to gauge CD4+ T cells directed against MTB.
An Agonist Antibody Specific for CD40 Induces Dendritic Cell Maturation and Promotes Autologous Anti‐tumour T‐cell Responses in an In vitro Mixed Autologous Tumour Cell/Lymph Node Cell Model
Authors: Hunter T. B. , Alsarraj M. , Gladue R. P. , Bedian V. , Antonia S. J.
Published: May 2007

CD40‐mediated interactions play an important role in the response to a variety of diseases, including cancer. Engagement of CD40 on antigen‐presenting cells, namely dendritic cells (DC), by CD40L leads to maturation and up‐regulation of co‐stimulatory molecules B7.1 and B7.2 (CD80 and CD86). These molecules are requisite to subsequent antigen‐specific activation of T cells. T‐cell activation is a critical aspect of specific anti‐tumour immune responses that have become the focus of a variety of cancer immunotherapy approaches. Clinical trials involving immunologic interventions have shown clinical responses confirming that the immune system can be harnessed for the treatment of cancer. However, the clinical response rate has been low, signifying the need for new immunotherapeutic strategies. To this end, an agonist antibody specific for CD40, CP‐870,893, has been developed. A fully autologous mixed tumour cell/lymph node cell model was utilized to demonstrate that CP‐870,893 promotes the responsiveness of lymph node‐derived T cells to autologous tumour. Specifically, T cells from the tumour‐draining lymph nodes are not responsive to autologous tumour cells; however, in the presence of CP‐870,893, this unresponsiveness is reversed, as indicated by lymph node cell proliferation and cytokine secretion. Monocyte‐derived DC treated with CP‐870,893 consistently display a mature phenotype: up‐regulation of CD80, CD83, CD86 and HLA‐DR expression, increased Mip1α and IL‐12 secretion, and the loss of exogenous antigen‐presenting capability subsequent to treatment with the antibody. These data indicate that CP‐870,893 binds to and activates DC, ultimately driving a specific anti‐tumour T‐cell response.
Epitope Mapping of Series of Monoclonal Antibodies Against the Hepatocellular Carcinoma‐associated Antigen HAb18G/CD147
Authors: Ku X.‐M. , Liao C.‐G. , Li Y. , Yang X.‐M. , Yang B. , Yao X.‐Y. , Wang L. , Kong L.‐M. , Zhao P. , Chen Z.‐N.
Published: May 2007

The hepatocellular carcinoma‐associated antigen HAb18G/CD147, a member of CD147 family, could promote tumour invasion and metastasis via inducing the secretion of matrix metalloproteinases (MMP). Anti‐CD147 monoclonal antibodies (MoAb) have exhibited obvious inhibitory effect on MMP induction. However, none of the epitopes of these MoAb has been reported. We previously prepared five MoAb against HAb18G/CD147, named HAb18, 3B3, 1B3, 5A5 and 4D2. To map the epitopes of these MoAb, a series of truncated fragments of extracellular region of HAb18G/CD147 was expressed in Escherichia coli and the MoAb‐binding affinity to these fragments was examined with an enzyme‐linked immunosorbent assay and Western blot. The residues 39LTCSLNDSATEV50, 36KILLTCS42 and 22AAGTVFTTVEDL33 were determined to be the epitopes of HAb18, 3B3 and 1B3, respectively, which were further proved by a dot‐blot analysis with synthesized peptides and bioinformatics epitope prediction. The binding regions of MoAb 5A5 and 4D2 were located at residues E120–R203. Then we constructed and expressed full‐length HAb18G/CD147 and truncated HAb18G/CD147 without residues A22–V50 in COS‐7 cells. Gelatin zymography and Boyden chamber assay showed that the COS‐7 cells expressing truncated HAb18G/CD147 failed to induce MMP production and enhance the cells’ invasive potential, compared with the cells expressing full‐length HAb18G/CD147. Taken together with the obviously inhibitory effects of HAb18 on the function of full‐length HAb18G/CD147, these findings suggest that residues 22AAGTVFTTVEDLGSKILLTCSLNDSATEV50 may play a critical role in the functions of HAb18G/CD147 on MMP secretion and tumour invasion. These key residues can be used as potential drug target in cancer therapy.
The Influence of MHC Class II Molecules Containing the Rheumatoid Arthritis Shared Epitope on the Immune Response to Aggrecan G1 and Its Peptides
Authors: Brintnell W. , Bell D. A. , Hill J. A. , Jevnikar A. M. , Sette A. , Sidney J. , Doege K. , Cairns E.
Published: May 2007

Aggrecan has been implied as an autoantigen in rheumatoid arthritis (RA). Immunization with aggrecan induces arthritis in BALB/c (H‐2d) mice but not in other strains of mice [e.g. C57BL/6 (H‐2b)]. In humans, the strongest genetic association with RA is to the shared epitope (SE), and aggrecan peptides are predicted to bind to the SE. Therefore, we hypothesized that C57BL/6 mice transgenic (tg) for the RA SE (DR4 tg mice) may be susceptible to aggrecan‐induced arthritis. C57BL/6 and DR4 tg mice were immunized with a mixture of SE‐binding aggrecan peptides and tested for immune responses to the corresponding peptides as well as aggrecan. Sustained T‐ and B‐cell immune responses to aggrecan and several of its peptides were detected in DR4 tg mice. C57BL/6 mice showed only transient T‐cell responses to different immunizing peptides and little B‐cell response. Therefore, an immune response to peptides of aggrecan can be induced experimentally in DR4 tg mice as anticipated from the predicted and actual binding affinities of these peptides for the RA SE. Failure to induce arthritis in these DR4 tg mice may be due to a lack of appropriate non‐MHC genes.
Genotypic characterization of Porphyromonas gingivalis isolated from Swedish patients with periodontitis and from periodontal abscesses
Authors: Yoshino T. , Laine M. L. , Van Winkelhoff A. J. , Dahlén G.
Published: May 2007

Introduction:  A significant genetic polymorphism has been shown for Porphyromonas gingivalis isolates from different geographical areas. It is, however, possible that genetic similarities can be found among isolates obtained from a more specific population. The aim of the present study was to evaluate genetic heterogeneity among P. gingivalis isolates obtained from Swedish subjects with chronic periodontitis and from periodontal abscess lesions.

Methods:  A total of 78 P. gingivalis strains, including 55 fresh clinical isolates obtained from 52 Swedish periodontitis subjects, eight isolates from eight Swedish periodontal abscess subjects and 15 reference strains, were subjected to amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) genotyping assays.

Results:  A total of 62 AFLP genotypes and 70 RAPD genotypes were identified among the 78 P. gingivalis strains. Forty‐six strains were clustered at 70% similarity level into 15 clusters. Six identical RAPD genotypes were identified among the strains. The AFLP/RAPD profiles were compared for identical genotypes. A total of 56 AFLP/RAPD genotypes were found. Four pairs of identical AFLP/RAPD genotypes were found for two strains obtained from two different periodontal pockets each of four subjects. Interestingly, two strains showed an RAPD/AFLP genotype, which was identical to the type strain W83.

Conclusion:  The present study demonstrated that Swedish P. gingivalis isolates exhibit a wide variety of genotypes with only a weak clustering pattern. No predominant genotype at the whole chromosomal DNA level was present among Swedish P. gingivalis strains.
Identification of periodontopathic bacteria in gingival tissue of Japanese periodontitis patients
Authors: Thiha K. , Takeuchi Y. , Umeda M. , Huang Y. , Ohnishi M. , Ishikawa I.
Published: May 2007

Introduction:  The identification of invading periodontopathic bacteria in tissues is important to determine their role in the pathogenesis of periodontal disease. The objective of this study was to identify periodontopathic bacteria in diseased gingival tissue of periodontitis patients.

Methods:  Subgingival plaque and gingival tissue were collected from 32 generalized chronic periodontitis (CP), 16 generalized aggressive periodontitis (GAgP) and eight localized aggressive periodontitis (LAgP) patients. Detection frequencies and quantities of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Tannerella forsythensis were investigated by polymerase chain reaction. The prevalences of Streptococcus oralis and Streptococcus sobrinus were also examined and the distribution of A. actinomycetemcomitans serotypes was observed.

Results:  P. gingivalis and T. forsythensis were detected in approximately 70% of tissue samples and 50% of plaque samples in the three periodontitis groups. Prevalence of A. actinomycetemcomitans in tissue samples was higher in the LAgP (63%) group than in either the CP (16%) or the GAgP (38%) group. A. actinomycetemcomitans serotype c was detected in 50% of LAgP patients. Detection frequencies of S. oralis and S. sobrinus were markedly low in both plaque and tissue samples from all three periodontitis groups. Amounts of P. gingivalis, A. actinomycetemcomitans and T. forsythensis in the tissue samples were not different among the three periodontitis groups.

Conclusion:  P. gingivalis, A. actinomycetemcomitans and T. forsythensis can localize in diseased gingival tissue and may be involved in periodontal tissue destruction. Serotype c is the predominant serotype of A. actinomycetemcomitans in Japanese LAgP patients.
Effects of Exogenous Glutathione on the Quality of Chilled Bull Semen
Authors: Munsi MN , Bhuiyan MMU , Majumder S , Alam MGS
Published: May 2007

Contents

Glutathione (GSH) at concentrations of 0.0 (control), 0.5, 1.0, 2.0 and 3.0 mM was added to chilled bull semen to determine its effects on the keeping quality of semen used for artificial insemination (AI). The semen was preserved with egg yolk citrate extender. All samples were stored at 4–8°C for 5 days. Sperm motility and proportion of abnormal acrosome were assessed daily. Sperm motility was significantly (p < 0.01) higher in the semen treated with 0.5 mM glutathione than in untreated semen on each day. The optimum sperm motility (≥50%) for AI was retained significantly (p < 0.01) for 3 days in 0.0, 0.5, 1.0 and 2.0 mM glutathione treated semen, whereas in 0.3 mM glutathione‐treated semen, sperm motility was 46.8% for 3 days. Acrosomal damage was significantly (p < 0.01) reduced after addition of 0.5 mM GSH in the preserved semen. Bull semen can be preserved in chilled condition for 5 days with 0.5 mM GSH with sperm motility above 40% and 12% acrosome abnormality.
Superovulatory Ovarian Response in Mangalica Gilts is Not Influenced by Feeding Level
Authors: Egerszegi I , Hazeleger W , Rátky J , Sarlós P , Kemp B , Bouwman E , Solti L , Brüssow K‐P
Published: May 2007

Contents

The aim of the study was to compare how different feeding levels affect the ovarian potential of follicular development and oocyte maturation in response to superovulatory treatment in native Mangalica (M, n = 17) compared with Landrace (L, n = 20) pigs. Gilts of both breeds were fed high‐energy (HI–2.5 kg) or low‐energy (LO – 1.25 kg) feed during oestrus synchronization (15 days of Regumate® feeding) till the time of oocyte aspiration (Day 6 after Regumate®). Follicular growth was stimulated by the administration of 1000 IU equiue choriou gonadotropiu (eCG) 24 h after Regumate® treatment, and ovulation was induced by injection of 750 IU human choriou gonadotropiu (hCG) 80 h after eCG adminstration. Ultrasound (US) investigation was done three times (4–10 h before, and 40–44 and 72–74 h after eCG administration) for the observation of follicular development. Oocyte and follicular fluid (FF) were collected endoscopically 34 h after hCG injection. Cumulus–oocyte complexes were evaluated, their morphology determined, and thereafter fixed and stained for chromatin evaluation. Oocytes were classified as meiosis‐resumed (germinal vesicle breakdown, diakinesis, metaphase I to anaphase I) or matured (telophase I and metaphase II). FF concentrations of oestradiol and progesterone were measured by validated radioimmunoassays. In L gilts, differences were observed between HI and LO in the number of preovulatory follicles (32.3 ± 10.5 vs 17.1 ± 12.3, p < 0.05), but not in M (25.3 ± 2.9 vs 28.8 ± 7.3, p > 0.05). Initial follicular growth was not affected by feeding levels; however, preovulatory follicle size was larger in M (7.1 ± 0.9 and 6.9 ± 1.1 mm vs 5.7 ± 0.7 and 5.5 ± 0.8 mm; p < 0.05). No differences were obtained with relation to mature chromatin configuration in both breeds (L gilts: HI – 70% and LO–67% vs M gilts: HI – 67% and LO – 63%). A twofold higher oestradiol concentration was detected in FF of HI‐M and LO‐M (29.6 ± 6.8 and 30.9 ± 10.3 ng/ml respectively) compared with that of L (16.9 ± 9.7 and 17.9 ± 3.6 ng/ml, respectively; p < 0.05). The mean FF progesterone level was nearly fivefold higher in M (2020.4 ± 1056 and 1512.2 ± 1121.8 ng/ml) compared with L (386.2 ± 113.7 and 298.8 ± 125.9 ng/ml, p < 0.05). The results indicate an influence of the feeding of altered energy on the number of recruitable preovulatory follicles in modern Landrace but not in native Mangalica breed. Moreover, the follicular steroid hormone milieu differs between Landrace and Mangalica gilts but not depending on feeding levels. Oocyte maturation was not affected by diet.
Ultrasonographic Characteristics of Ovulatory Follicles and Associated Endocrine Changes in Cyclic Ewes Treated with Medroxyprogesterone Acetate (MAP)‐releasing Intravaginal Sponges and Equine Chorionic Gonadotropin (eCG)
Authors: Liu X , Dai Q , Hart EJ , Barrett DMW , Rawlings NC , Pierson RA , Bartlewski PM
Published: May 2007

Contents

The aim of this study was to assess the ultrasonographic characteristics of ovulatory follicles in cyclic Western White Face ewes (December) that had received intravaginal sponges containing medroxyprogesterone acetate (MAP; 60 mg) for 12 days, with or without an injection of 500 IU of equine chorionic gonadotropin (eCG) at sponge removal. We hypothesized that quantitative echotextural attributes of the follicles in ewes treated only with MAP would differ from those in MAP/eCG‐treated ewes, reflecting the increased antral follicular growth and secretory function under eCG influence. Digital images of ovulatory follicles obtained at 0 and 24 h after MAP sponge removal and at 24 h before ovulation in the eCG‐treated (five ewes, 13 follicles) and control (six ewes, 9 follicles) animals, were subjected to computerized analyses. The mean diameter of ovulatory follicles increased (p < 0.001) 24 h after eCG treatment. The mean pixel intensity and heterogeneity of the follicular antrum (p < 0.001), as well as mean pixel intensity of the follicular wall and perifollicular ovarian stroma (p < 0.05), were greater in eCG‐treated animals compared with control ewes 24 h after sponge removal and at 24 h before ovulation. Mean serum concentrations of oestradiol‐17β tended to increase (p = 0.06) 24 h after eCG treatment and the eCG‐treated ewes exceeded (p < 0.05) control animals in progesterone concentrations from days 9–15 after ovulation. Our results support the hypothesis that large antral follicles in eCG‐treated ewes exhibit distinctive echotextural characteristics. Follicular image attributes in eCG‐treated ewes appear to be indicative of the changes in follicular morphology and secretory activity caused by the administration of the exogenous gonadotropin, which has both FSH‐ and LH‐like activities.
Antioxidants in Bovine Colostrum
Authors: Przybylska J , Albera E , Kankofer M
Published: May 2007

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Quality, content and properties of colostrum are crucial for the neonate and its further development. Due to essential differences between intrauterine and extrauterine environment, the neonate is exposed to oxidative stress conditions. Colostrum apart from nutrient and immunological components should contain antioxidative systems necessary for the protection against reactive oxygen species. This review describes available data on enzymatic and non‐enzymatic antioxidants in colostrum. Due to the fact that the literature concerning bovine colostrum is scanty, the information based on bovine mature milk determinations as well as other species is provided. Bovine colostrum is used not only by calves, but also for the production of hyperimmunized colostrum, medicines or feed supplements. Quality of colostrum influences quality of mature milk. This is another reason, except from health of neonate, why antioxidative properties of bovine colostrum are of special importance and require further detailed elucidation.
Synchrony of Ovulation and Follicular Dynamics in Merino Ewes Treated with GnRH in the Breeding and Non‐breeding Seasons
Authors: Reyna J , Thomson PC , Evans G , Maxwell WMC
Published: May 2007

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The effect of gonadotropin releasing hormone (GnRH) treatment on the time of ovulation and the occurrence of follicular dominance during the non‐breeding and breeding seasons (experiment 1), and on fertility after artificial insemination (AI) in the non‐breeding season (experiment 2), was examined in Merino ewes. Oestrus was synchronized in 40 nulliparous ewes (experiment 1; n = 20, in the non‐breeding and breeding seasons) and in 79 multiparous ewes (experiment 2) using intravaginal sponges and pregnant mare serum gonadotropin. Thirty six hours after sponge removal (SR), half the ewes were injected (i.m.) with 40 μg of synthetic GnRH and the remainder used as controls. GnRH improved the synchrony of ovulation compared with the controls in the breeding (SD = 2.8 vs 5.7 days, p = 0.04) but not the non‐breeding season (SD = 3.8 vs 4.4 days, p = 0.69), with ewes ovulating from 42 to 54 h (mean 50.4 ± 4.08 h) and 42–60 h (mean 54.4 ± 5.47 h) after SR for GnRH and control, respectively. For both treated and control ewes, ovulation occurred earlier in the non‐breeding than the breeding season (50.1 vs 54.6 h; p = 0.002). GnRH had no effect on follicular dominance, as assessed by divergence (D: the time the ovulatory follicle exceeded the average size of the other non‐ovulating follicles) or on the interval from D to ovulation (IDO). However, follicular dynamics differed between seasons. The mean follicle diameter increased at a faster rate up to 36 h after SR in the non‐breeding compared with the breeding season and then rapidly declined, compared with a later peak (42 h after SR) in mean follicular size during the breeding season. IDO was shorter in the non‐breeding than in the breeding season (26.7 ± 4.30 h vs 39.6 ± 4.53 h; p = 0.05). In experiment 2, ewes (n = 38 GnRH‐treated, n = 40 controls) were inseminated in the uterus by laparoscopy 42 h or 48 h after SR with frozen‐thawed sperm. The fertility of ewes treated with GnRH (nine of 39, 23%) was not different to the controls (eight of 38, 21%; p = 0.01). In conclusion the application of GnRH improved synchronization of ovulation but did not improve fertility rates after AI.
Effect of Insemination–Ovulation Interval and Addition of Seminal Plasma on Sow Fertility to Insemination of Cryopreserved Sperm
Authors: Abad M , Garcia JC , Sprecher DJ , Cassar G , Friendship RM , Buhr MM , Kirkwood RN
Published: May 2007

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In swine, the use of frozen‐thawed (FT) sperm for artificial insemination (AI) is limited because of poor sow fertility, possibly associated with a post‐thaw capacitation‐like status resulting in fewer fully viable sperm. Sow fertility to AI with FT sperm may improve with deeper deposition of sperm within the female tract, insemination very close to ovulation, or reversal of cryocapacitation by seminal plasma (SP). We performed two experiments to examine these suggestions. In experiment 1, 122 multiparous Yorkshire sows received 600 IU equine chorionic gonadotrophin at weaning and 5 mg pLH 80 h later to control time of ovulation. The predicted time of ovulation (PTO) was 38 h after pLH injection. Thereafter, sows were assigned on the basis of parity to a single AI of FT sperm at 2 h before PTO, or at 12 h before PTO, or FT sperm supplemented with 10% SP at 12 h before PTO. Control sows received fresh semen at 12 h before PTO. All semen doses were adjusted to 3 × 109live cells and deposited into the cervix. Experiment 2 employed 99 multiparous crossbred sows and repeated the treatments of experiment 1 except that all FT inseminations were intrauterine. In both experiments, farrowing rates were lower (p < 0.01) following FT inseminations with no effect of time of insemination or of supplemental SP. In experiment 1, litter size was smaller following FT insemination (p < 0.05), but no effect on litter size was evident in experiment 2. Supplemental SP had no effect on litter size in either experiment. The lack of effect of either SP or timing of FT insemination on sow fertility suggests that the non‐lethal sperm cryoinjury affecting fertility involves more than just cryocapacitation.
Comparison of Five Radioimmunoassay Systems for PAG Measurement: Ability to Detect Early Pregnancy in Cows
Authors: Ayad A , Sousa NM , Sulon J , Iguer‐Ouada M , Beckers JF
Published: May 2007

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This study was conducted to describe the minimum detection limit, reproducibility, accuracy, specificity and parallelism of different pregnancy‐associated glycoprotein radioimmunoassay (PAG‐RIA) systems: RIA‐497, RIA‐706, RIA‐780, RIA‐809 and RIA‐Pool. Their ability to distinguish between non‐pregnant and pregnant females at day 30 after artificial insemination (AI) was investigated. The antisera were raised in rabbits against different PAG preparations. All RIA systems proved to be sensitive, repeatable and accurate for measuring PAG concentrations. The dilutions of plasma samples taken at an early stage of pregnancy were found to be parallel to the standard curves. No cross‐reaction was observed with different carbohydrates, either with Pregnant Mare Serum Gonadotropin (PMSG) or human Chorionic Gonadotropin (hCG). The concentrations of PAG in pregnant females at day 30 after AI were shown to be higher with the use of antisera R#706, R#780, R#809 and Pool when compared with antiserum R#497. All the RIA systems gave 100% sensitivity and negative predictive values. On the other hand, the use of antisera R#780 and R#809 resulted in lower specificity and positive predictive values. The present study clearly shows that the ability of PAG‐RIA systems to diagnose pregnancy specifically at day 30 after AI can be improved by using a combination of antisera raised against different forms of PAG.
Operation Child‐ID: reunifying children with their legal guardians after Hurricane Katrina
Authors: Brandenburg Mark A. , Watkins Sue M. , Brandenburg Karin L. , Schieche Christoph
Published: May 2007

Children constitute a vulnerable population and special considerations are necessary in order to provide proper care for them during disasters. After disasters such as Hurricane Katrina, the rapid identification and protection of separated children and their reunification with legal guardians is necessary in order to minimise secondary injuries (i.e. physical and sexual abuse, neglect and abduction). At Camp Gruber, an Oklahoma shelter for Louisianans displaced by Hurricane Katrina, a survey tool was used to identify children separated from their guardians. Of the 254 children at the camp, 36 (14.2 per cent) were separated from their legal guardians. Answering ‘no’ to the question of whether the accompanying adult was the guardian of the child prior to Hurricane Katrina was a strong predictor (27.8 per cent versus 3.2 per cent) of being listed as ‘missing’ by the National Center for Missing and Exploited Children (NCMEC). All the children at Camp Gruber who were listed as ‘missing’ by the NCMEC were subsequently reunited with their guardians.
572. BERGENIA EMEIENSIS
Authors: Ogisu Mikinori , Rix Martyn
Published: May 2007

Summary Bergenia emeiensis C.Y. Wu ex J.T. Pan is illustrated and described. Its relationship to other Himalayan species is discussed, and instructions for its cultivation are given.
573. GERANIUM GLABERRIMUM
Authors: Wilford Richard , Walters Graham
Published: May 2007

Summary The rare Turkish endemic Geranium glaberrimum Boiss. & Heldr. is illustrated and described. Its close relatives, classification, distribution and cultivation are discussed.