bluuebunny/crossref_metadata_2025 huggingface dataset:
An in vitro antimicrobial comparison of miswak extract with commercially available non‐alcohol mouthrinses
Authors: Almas K , Skaug N , Ahmad I
Published: January 2005

The aim of this study was to assess the antimicrobial activity of eight commercially available mouthrinses and 50% miswak extract against seven microorganisms. Corsodyl, Alprox, Oral‐B advantage, Florosept, Sensodyne, Aquafresh Mint, Betadine and Emoform mouthrinses were used while 50% aqueous extract of miswak (Salvadora persica) was used against Streptococcus faecalis, Streptococcus pyogenis, Streptococcus mutans, Candida albicans, Staphylococcus aureus and Staphylococcus epidermidis. The ditch plate method was used to test the antimicrobial activity. Inhibition zones of microorganisms around ditches were measured in millimetres. Range, mean and standard deviations were used for comparison of antimicrobial activity. Mouthrinses containing chlorhexidine was with maximum antibacterial activity, while cetylpyridinium chloride mouthrinses were with moderate and miswak extract was with low antibacterial activity. Further research is needed for the substantivity of these mouthrinses and further in vivo/in vitro studies are needed using Biofilm model to substantiate present findings. Dental professionals must exercise caution and provide guidance in assisting their patients in making informed choices regarding their use of mouthrinses for clinical efficacy.
International employment in clinical practice: influencing factors for the dental hygienist
Authors: Abbott A , Barrow S‐Y , Lopresti F , Hittelman E
Published: January 2005

Purpose:  To assess demographics, job characteristics, geographical regions, resources and commitment, which influence dental hygienists seeking international clinical practice employment opportunities.

Methods:  Questionnaires were mailed to a convenience sample of members of the Dental Hygienists’ Association of the City of New York. Statistical analyses were conducted and frequency distributions and relationships between variables were calculated.

Results:  Seventy‐two percent of respondents reported that they are or may be interested in working overseas. Italy and Spain (67%) were the regions of most interest. Salary (65%) was cited as the most influencing factor in selection, whereas non‐compliance with the equivalency to Occupational Safety and Health Administration standards (74%) was the most frequently perceived barrier. Multiple language fluency was statistically significant (0.003) regarding interest in overseas employment.

Conclusion:  Policy makers, employers and educators need to be aware of these findings should recruitment be a possibility to render urgently needed oral hygiene care in regions where there is a perceived shortage of dental hygienists.
Mapping an appropriate health promotion approach for crèches in an informal settlement
Authors: Brijlal P , Gordon N
Published: January 2005

Background:  People living in informal settlements in South Africa experience the double burden of poverty and ill health. Wallacedene, an informal settlement was highlighted in the media as being a socially and otherwise deprived community, with many accompanying health problems.

Aim:  It was against this background that this study was conducted to gain a better understanding of the health and oral health status of children attending crèches in Wallacedene. It was designed to inform the mapping of an appropriate approach to develop a health promotion programme for crèches.

Method:  Baseline data were collected through oral and general health examinations, site observations, a structured questionnaire and interviews with key people working with the children at two crèches.

Results:  The results indicate poor oral and general health. Gingival inflammation (82.8%), caries (81.5%), and moderate to abundant plaque deposits (95.7%), fungal infections (33.9%), runny nose (51.4%), lymphadenopathy (45.7%) and itchy skin (5.7%) were found. Caregivers were not well informed about oral health. However, they were enthusiastic to engage in new interventions. The community was impoverished; public health interventions were limited with minimal resources such as health centres and voluntary service providers. The limited resources were not coordinated and did not adequately address the health and educational needs of the children.

Conclusion:  A multi‐sectoral approach focusing on community development is an appropriate approach to address the needs of crèche children in this community.
A stylized computational model of the head for the reference Japanese male
Authors: Yamauchi M. , Ishikawa M. , Hoshi M.
Published: January 2005

Computational models of human anatomy, along with Monte Carlo radiation transport simulations, have been used by Snyder et al. [MIRD Pamphlet No. 5, revised (The Society of Nuclear Medicine, New York, 1978)], Cristy and Eckerman [ORNL/TM‐8381/VI, Oak Ridge National Laboratory, Oak Ridge, TN (1987)] and Zubal et al. [Med. Phys. 21, 299–302 (1994)] to estimate internal organ doses from internal and external radiation sources. These were created using physiological data from Caucasoid subjects but not from other races. There is a need for research to determine whether the obvious differences from the Caucasoid anatomy make these models unsuitable for estimating the absorbed dose in other races such as the Mongoloid. We used the cranial region of the adult Japanese male to represent the Mongoloid race. This region contains organs that are highly sensitive to radiation. The cranial region of a physical phantom produced by KYOTO KAGAKU Co., LTD. using numerical data from a Japanese Reference Man [Tanaka, Nippon Acta. Radiol. 48, 509–513 (1988)] was used to supply the data for the geometry of a stylized computational model. Our computational model was constructed with equations rather than voxel‐based, in order to deal with as small a number of parameters as possible in the computer simulation experiment. The accuracy of our computational model was checked by comparing simulated experimental results obtained with MCNP4C with actual doses measured with thermoluminescence dosimeters (TLDs) inside the physical phantom from which our computational model was constructed. The TLDs, whose margin of error is less than , were arranged at six positions. Co‐60 was used as the radiation source. The irradiated dose was 2 Gy in terms of air kerma. In the computer simulation experiments, we used our computational model and Cristy's computational model, whose component data are those of the tissue substitute materials and of the human body as published in ICRU Report 46. The observed absorbed dose values (Gy) at all six points were calculated as the percentage difference between MCNP4C simulation and the TLDs. In our computational model, the average values of all the percentage differences were (tissue substitute materials) and (ICRU Report 46), respectively. In Cristy's model, the corresponding values were (tissue substitute materials) and (ICRU Report 46), respectively. Considering the margin of error in the radiation sensitivity of the TLDs, this study validates our computational model as a test object for radiation dosimetry studies.
Reduction of exposure of newborns and caregivers to very high electromagnetic fields produced by incubators
Authors: Bellieni C. V. , Bagnoli F. , Pinto I. , Stacchini N. , Buonocore G.
Published: January 2005

The aims of this paper is to measure whether ferromagnetic panels sufficiently reduce the high electromagnetic fields (EMF) to which newborns are exposed in incubators and to which caregivers are exposed when working near the incubators. We measured EMF at mattress level in three neonatal incubators with and without ferromagnetic panels between the electric motor and the mattress. We then measured the EMF at the level of the maximum emission point for caregivers, i.e., near the display panel. The ferromagnetic panels were (a) 5 mm thick iron, (b), (c), (d) respectively, one, two, and three sheets of 0.3 mm thick mu‐metal. The weight of iron sheet was , and mu‐metal . The use of the ferromagnetic panels significantly reduced the EMF. No significant difference in attenuation was recorded using one, two, or three sheets of mu‐metal, or a single sheet of iron. One, two, and three sheets of mu‐metal reduced EMFs by 77%, 82%, and 84.3%, respectively; the reduction with iron was 80%. EMF values measured in incubators were higher than those to which the general population is exposed. The use of ferromagnetic panels significantly reduces the level of EMFs to which neonates and caregivers are exposed.
Rest period duration of the coronary arteries: Implications for magnetic resonance coronary angiography
Authors: Shechter Guy , Resar Jon R. , McVeigh Elliot R.
Published: January 2005

Magnetic resonance (MR) and computed tomography coronary imaging is susceptible to artifacts caused by motion of the heart. The presence of rest periods during the cardiac and respiratory cycles suggests that images free of motion artifacts could be acquired. In this paper, we studied the rest period (RP) duration of the coronary arteries during a cardiac contraction and a tidal respiratory cycle. We also studied whether three MR motion correction methods could be used to increase the respiratory RP duration. Free breathing x‐ray coronary angiograms were acquired in ten patients. The three‐dimensional (3D) structure of the coronary arteries was reconstructed from a biplane acquisition using stereo reconstruction methods. The 3D motion of the arterial model was then recovered using an automatic motion tracking algorithm. The motion field was then decomposed into separate cardiac and respiratory components using a cardiac respiratory parametric model. For the proximal‐to‐middle segments of the right coronary artery (RCA), a cardiac RP ( 3D displacement) of was measured at end systole (ES), and in mid‐diastole (MD). The cardiac RP was at ES and at MD for the proximal of the left coronary tree. At end expiration, the respiratory RP (in percent of the respiratory period) was for the RCA and for the left coronary tree. Left coronary respiratory RP ( 3D displacement) increased with translation (32% of the respiratory period), rigid body (51%), and affine (79%) motion correction. The RCA respiratory RP using translational (27%) and rigid body (33%) motion correction were not statistically different from each other. Measurements of the cardiac and respiratory rest periods will improve our understanding of the temporal and spatial resolution constraints for coronary imaging.
Annoyance with aircraft noise in local recreational areas and the recreationists’ noise situation at home
Authors: Krog Norun Hjertager , Engdahl Bo
Published: January 2005

Few socioacoustic studies have examined the effect of noise on outdoor recreationists. Most studies concentrate on one setting of the everyday life of a noise-exposed population, which mainly has been the residential setting. This article relates annoyance with aircraft noise in outdoor recreational areas to the recreationists’ noise situation at home. In conjunction with the relocation of the main airport of Norway in 1998, field studies were conducted before and after the change in one area near the old airport (1930 survey respondents), and one area near the new airport (1001 survey respondents). Multivariate linear regression analyses of the relationship between annoyance and aircraft noise exposure (LAeq for the aircraft events) in the recreational areas were conducted, controlled for noise annoyance at home, or aircraft noise exposure at home, the situation (before/after the change), context- and demographic variables. People more highly annoyed at home tended to be more annoyed than others while in the recreational areas. A significant effect of aircraft noise exposure at home on annoyance in the recreational setting was not found. More research is warranted regarding the relationship between noise exposure at home and outdoor recreational demands.
Iteration of transmit-beam aberration correction in medical ultrasound imaging
Authors: Måsøy Svein-Erik , Varslot Trond , Angelsen Bjørn
Published: January 2005

Simulations of iterative transmit-beam aberration correction using a time-delay and amplitude filter have been performed to study the convergence of such a process. Aberration in medical ultrasonic imaging is usually modeled by arrival-time and amplitude fluctuations concentrated on the transducer array. This is an approximation of the physical aberration process, and may be applied to correct the transmitted signal using a time-delay and amplitude filter. Estimation of such a filter has proven difficult in the presence of severe aberration. Presented here is an iterative approach, whereby a filter estimate is applied to correct the transmit-beam. This beam induces acoustic backscatter better suited for arrival-time and amplitude estimation, thus facilitating an improved filter estimate. Two correlation-based methods for estimating arrival-time and amplitude fluctuations in received echoes from random scatterers were employed. Aberration was introduced using eight models emulating aberration produced by the human abdominal wall. Results show that only a few iterations are needed to obtain corrected transmit-beam profiles comparable to those of an ideal aberration correction filter. Furthermore, a previously developed focusing criterion is found to quantify the convergence accurately.
Fine-tuning molecular acoustic models: sensitivity of the predicted attenuation to the Lennard–Jones parameters
Authors: Petculescu Andi G. , Lueptow Richard M.
Published: January 2005

In a previous paper [Y. Dain and R. M. Lueptow, J. Acoust. Soc. Am. 109, 1955 (2001)], a model of acoustic attenuation due to vibration-translation and vibration-vibration relaxation in multiple polyatomic gas mixtures was developed. In this paper, the model is improved by treating binary molecular collisions via fully pairwise vibrational transition probabilities. The sensitivity of the model to small variations in the Lennard–Jones parameters—collision diameter (σ) and potential depth (ε)—is investigated for nitrogen-water-methane mixtures. For a N2(98.97%)-H2O(338 ppm)-CH4(1%) test mixture, the transition probabilities and acoustic absorption curves are much more sensitive to σ than they are to ε. Additionally, when the 1% methane is replaced by nitrogen, the resulting mixture [N2(99.97%)-H2O(338 ppm)] becomes considerably more sensitive to changes of σwater. The current model minimizes the underprediction of the acoustic absorption peak magnitudes reported by S. G. Ejakov et al. [J. Acoust. Soc. Am. 113, 1871 (2003)].
Perceptual differences between low and high rates of stimulation on single electrodes for cochlear implantees
Authors: Landsberger David M. , McKay Colette M.
Published: January 2005

Previous research has shown that increases in the rate of stimulation on a single electrode yield changes in pitch perception until the rate is increased beyond a given critical rate, after which changes in rate are only perceived as changes in loudness. The critical rate beyond which a rate increase no longer elicits a pitch change in most subjects is approximately 300 Hz, although a small number of subjects have been observed to have critical rates up to approximately 1000 Hz. In this article, we sought to determine if increasing the rate of stimulation beyond the critical rate (up to 12.8 kHz) would eventually result in new changes of perception (other than loudness.) Our data replicate the previously observed results that rates between approximately 300 and 1500 Hz are indistinguishable from each other. However, we observed the finding that a rate of stimulation well above the critical rate (starting between 1500 Hz and 12.8 kHz, depending on electrode and subject) can elicit changes in perception. The perceptual differences between these high rates were sometimes but not always labeled as pitch changes. This phenomenon needs further research to assess its potential relevance to speech perception using high rates of stimulation.
Ambient noise cross correlation in free space: Theoretical approach
Authors: Roux Philippe , Sabra Karim G. , Kuperman W. A. , Roux Andre
Published: January 2005

It has been experimentally demonstrated that the Green’s function between two points could be recovered using the cross-correlation function of the ambient noise measured at these two points. This paper investigates the theory behind this result in the simple case of a homogeneous medium with attenuation.
Estimation of frequency-averaged loss factors by the power injection and the impulse response decay methods
Authors: Bloss Brandon C. , Rao Mohan D.
Published: January 2005

This paper describes a comparison, both analytically and experimentally, between two widely used loss factor estimation techniques frequently used in statistical energy analysis. Analytical models of simple spring/mass/damper systems were created to compare frequency-averaged loss factor values from the single subsystem power injection method and the impulse response decay method. The parameters of the analytical models were varied to study the effects of the total number of modes, amount of damping, location of modes within frequency bands, and the width of the frequency bands on loss factor estimation. The analytical study shows that both methods give accurate loss factor values as long as the damping values remain realistic for linear systems and at least one modal resonance is present in each frequency band. These analytical results were verified experimentally by measuring the loss factors of simple steel plates, with and without damping treatments applied.
Antigenic and Immunogenic Investigation of B-Cell Epitopes in the Nucleocapsid Protein of Peste des Petits Ruminants Virus
Authors: Choi Kang-Seuk , Nah Jin-Ju , Ko Young-Joon , Kang Shien-Young , Yoon Kyoung-Jin , Jo Nam-In
Published: January 2005

Attempts were made to identify and map epitopes on the nucleocapsid (N) protein of peste des petits ruminants virus (PPRV) (Nigeria75/1 strain) using seven monoclonal antibodies (MAbs) and deletion mutants. At least four antigenic domains (A-I, A-II, C-I, and C-II) were identified using the MAbs. Domains A-I (MAb 33-4) and A-II (MAbs 38-4, P-3H12, and P-13A9) were determined to be located on the amino-terminal half (amino acids [aa] 1 to 262), and domains C-I (P-14C6) and C-II (P-9H10 and P-11A6) were within the carboxy-terminal region (aa 448 to 521). Nonreciprocal competition between A-II MAbs and MAbs to C-I and C-II domains was observed, indicating that they may be exposed on the surface of the N protein and spatially overlap each other. Blocking or competitive enzyme-linked immunosorbent assay studies using PPRV serum antibodies revealed that epitopes on the domains A-II and C-II were immunodominant, whereas those on the domains A-I and C-I were not. The competition between MAb and rinderpest virus (RPV) serum antibodies raised against RPV strain LATC was found in two epitopes (P-3H12 and P-13A9) on the domain A-II, indicating that these epitopes may cause cross-reactivity between PPRV and RPV. Identification of immunodominant but PPRV-specific epitopes and domains will provide the foundation in designing an N-protein-based diagnostic immunoassay for PPRV.
Cytokine Responses to Stimulation of Whole Blood from Patients with Buruli Ulcer Disease in Ghana
Authors: Westenbrink B. Daan , Stienstra Ymkje , Huitema Minke G. , Thompson William A. , Klutse Erasmus O. , Ampadu Edwin O. , Boezen H. Marike , Limburg Piet C. , van der Werf Tjip S.
Published: January 2005

Buruli ulcer disease (BUD), caused byMycobacterium ulcerans, follows an indolent course of initial progression to ulceration accompanied by extensive tissue damage. It has been suggested that healing disease stages are accompanied by a protective immune response. We hypothesized that interleukin-4 (IL-4)- or IL-10-induced downregulation of Th-1 responses plays a key role in the progression of early BUD and that healing is accompanied by an augmented Th-1 response. Gamma interferon (IFN-γ), IL-4, and IL-10 responses were measured after in vitro stimulation with phytohemagglutinin (PHA) and tuberculin purified protein derivative (PPD) of whole blood from 39 (23 early- and 16 late-stage) BUD patients and 39 healthy control subjects in Ghana. Additionally, 30 patients with active or treated tuberculosis (TB) serving as PPD-responsive positive controls were studied. Early-stage BUD patients produced significantly lower levels of IFN and IFN-γ/IL-4 ratios compared to late-stage BUD patients after PHA stimulation. Compared to that of controls, IFN-γ production after tuberculin stimulation was significantly higher in late-stage but not in early-stage BUD patients (P= 0.009). IL-10 and IL-4 levels did not differ between BUD patients and controls, although active TB patients had significantly higher IL-10 production levels than did treated TB patients. Multivariate analysis showed no confounding factors. In conclusion, Th-1 down regulation in early BUD appears to reverse in later stages of BUD, although an association with IL-10 or IL-4 production does not emerge from our data. Here we show differences in Th-1-type cytokine production between early- and late-stage BUD that might reflect an improved immune defense over time.
Effects of Thalidomide on IntracellularMycobacterium lepraein Normal and Activated Macrophages
Authors: Tadesse A. , Shannon E. J.
Published: January 2005

Thalidomide is an effective drug for the treatment of erythema nodosum leprosum (ENL). ENL is an inflammatory reaction that may occur in multibacillary leprosy patients. Its cause(s) as well as the mechanism of thalidomide in arresting this condition are not fully understood. It has been suggested that ENL is an immune complex-mediated hypersensitivity precipitated by the release ofMycobacterium lepraefrom macrophages. The released antigen may complex with precipitating antibodies, initiating complement fixation and the production of inflammatory cytokines like tumor necrosis factor alpha (TNF-α). Thalidomide has been shown in vitro to reduce antigen- or mitogen-activated macrophage production of TNF-α. We investigated if thalidomide could also influence the viability of intracellularM. leprae. Mouse peritoneal macrophages were infected withM. leprae, activated with gamma interferon and endotoxin, or nonactivated, and treated with thalidomide. Intracellular bacilli were recovered, and metabolic activity was assessed by a radiorespirometric procedure. Thalidomide did not possess antimicrobial action againstM. lepraein normal and activated host macrophages. This suggests that thalidomide does not retard the release of mycobacterial antigens, a possible prelude or precipitating factor for ENL. A distinct sequence of events explaining the mechanism of action for thalidomide's successful treatment of ENL has yet to be established.
Efficacy of Several Serological Tests and Antigens for Diagnosis of Bovine Brucellosis in the Presence of False-Positive Serological Results Due to
Yersinia enterocolitica
O:9
Authors: Muñoz P. M. , Marín C. M. , Monreal D. , González D. , Garin-Bastuji B. , Díaz R. , Mainar-Jaime R. C. , Moriyón I. , Blasco J. M.
Published: January 2005

Yersinia enterocolitica
O:9 bears a smooth lipopolysaccharide (S-LPS) of
Brucella
sp. O-chain A + C/Y epitopic structure and is a cause of false-positive serological reactions (FPSR) in standard tests for cattle brucellosis.
Brucella
S-LPS, cross-reacting S-LPSs representing several O-chain epitope combinations,
Brucella
core lipid A epitopes (rough LPS),
Brucella abortus
S-LPS-derived polysaccharide, native hapten polysaccharide, rough LPS group 3 outer membrane protein complexes, recombinant BP26, and cytosolic proteins were tested in enzyme-linked immunosorbent assays (ELISA) and precipitation tests to detect cattle brucellosis (sensitivity) and to differentiate it from FPSR (specificity). No single serological test and antigen combination showed 100% sensitivity and specificity simultaneously. Immunoprecipitation tests with native hapten polysaccharide, counterimmunoelectrophoresis with cytosolic proteins, and a chaotropic ELISA with
Brucella
S-LPS were 100% specific but less sensitive than the Rose Bengal test, complement fixation, and indirect ELISA with
Brucella
S-LPSs and native hapten or S-LPS-derived polysaccharides. A competitive ELISA with
Brucella
S-LPS and M84 C/Y-specific monoclonal antibody was not 100% specific and was less sensitive than other tests. ELISA with
Brucella suis
bv. 2 S-LPS (deficient in C epitopes),
Escherichia hermannii
S-LPSs [lacking the contiguous α-(1-2)-linked perosamine residues characteristic of
Y. enterocolitica
S-LPS], BP26 recombinant protein, and
Brucella
cytosolic fractions did not provide adequate sensitivity/specificity ratios. Although no serological test and antigen combination fully resolved the diagnosis of bovine brucellosis in the presence of FPSR, some are simple and practical alternatives to the brucellin skin test currently recommended for differential diagnosis.
Serological Specificities of Murine Hybridoma Monoclonal Antibodies againstNeisseria meningitidisSerogroups B, C, Y, and W135 and Evaluation of Their Usefulness as Serogrouping Reagents by Indirect Whole-Cell Enzyme-Linked Immunosorbent Assay
Authors: Tsang Raymond S. W. , Zollinger Wendell D.
Published: January 2005

Murine hybridoma monoclonal antibodies (MAbs) were produced against the capsular antigens of serogroups B, C, Y, and W135 meningococci. Each serogroup-specific MAb reacted with the extracted capsular polysaccharide from its homologous serogroup only and did not react with capsules from the other three serogroups. The application of these MAbs for serogroup identification of meningococci was demonstrated by their abilities to correctly identify 183 clinical isolates of 185 meningococci recovered from individual invasive meningococcal disease (IMD) patients during routine surveillance in 2002. The remaining two meningococci were identified by PCR grouping as C in one case and Y in another, but neither isolate was positive by bacterial agglutination using rabbit antisera or by enzyme-linked immunosorbent assay using MAbs. The specificities of the anti-Y and anti-W135 MAbs were further assessed by tests with 37 serogroup W135 and 106 serogroup Y meningococci recovered from IMD cases during 1999 to 2001 and 2003. All 143 meningococci except one serogroup Y isolate were correctly identified by positive reactions with the corresponding MAbs that identified their homologous serogroups. The single serogroup Y isolate was received as nonagglutinable and tested as negative with both rabbit anti-Y antiserum and anti-Y MAb but was positive for the serogroup Y-specificsiaDgene. The advantage of using MAbs for serogrouping of meningococci is discussed.
Assessment of Antibody Response Elicited by a 7-valent Pneumococcal Conjugate Vaccine in Pediatric Human Immunodeficiency Virus Infection
Authors: Tarragó David , Casal Julio , Ruiz-Contreras Jesús , Ramos J. Tomás , Rojo Pablo , Snippe Harm , Jansen Wouter T. M.
Published: January 2005

We investigated antibody responses against pneumococci of serotypes 6B, 14, and 23F in 56 children and adolescents with perinatal human immunodeficiency virus (HIV) infection who were vaccinated with 7-valent pneumococcal conjugate vaccine. Overall immune responses differed greatly between serotypes. Correlation coefficients between immunoglobulin G (IgG) measured by enzyme-linked immunosorbent assay (ELISA) and functional antibodies measured by a flow cytometry opsonophagocytosis assay (OPA) varied with serotype and time points studied. After 3 months of administering a second PCV7 dose we got the highest correlation (with significant
r
values of 0.754, 0.414, and 0.593 for serotypes 6B, 14, and 23F, respectively) but no significant increase in IgG concentration and OPA titers compared to the first dose. We defined a responder to a serotype included in the vaccine with two criteria: frequency of at least twofold OPA and ELISA increases for each serotype and frequency of conversion from negative to positive OPA levels. Responders varied from 43.9% to 46.3%, 28.5% to 50.0%, and 38.0% to 50.0% for serotypes 6B, 14, and 23F, respectively, depending on the response criterion. The present research highlights the importance of demonstrating vaccine immunogenicity with suitable immunological endpoints in immunocompromised patients and also the need to define how much antibody is required for protection from different serotypes, since immunogenicity differed significantly between serotypes.
Induction of Mucosal B-Cell Memory by Intranasal Immunization of Mice with Respiratory Syncytial Virus
Authors: Valosky Janine , Hishiki Haruka , Zaoutis Theoklis E. , Coffin Susan E.
Published: January 2005

The capacity of live or inactivated respiratory syncytial virus (RSV) to induce B-cell memory in respiratory-associated lymphoid tissues of mice was examined. Eight weeks after primary inoculation with either live or inactivated RSV, adult BALB/c mice were challenged with 4 × 105PFU of RSV. Protection from viral shedding and mucosal production of RSV-specific antibodies were examined at various time points after challenge. We found that primary immunization with live, but not inactivated, RSV induced complete and durable protection upon challenge within the upper and lower respiratory tract. Also, primary immunization with live, but not inactivated, RSV enhanced the production of mucosal RSV-specific immunoglobulin A (IgA) upon challenge. Secondary mucosal IgA responses were characterized by (i) the early production of mucosal IgA by B cells that reside in organized nasal-associated lymphoid tissues, cervical lymph nodes, and bronchial lymph nodes, and (ii) the subsequent production of RSV-specific IgA by mucosal effector tissues, such as the tracheal lamina propria and lung. These findings suggest that primary infection of mice with live RSV might induce mucosal IgA-committed memory B cells. A greater understanding of the characteristics of RSA-specific mucosal memory B cells may facilitate the development of an RSV vaccine.
Antibody Responses of Pigs to Defined E
rns
Fragments after Infection with Classical Swine Fever Virus
Authors: Lin Min , Trottier Erin , Pasick John
Published: January 2005

Antibody responses of pigs to defined E
rns
fragments, after classical swine fever virus (CSFV) infection, were studied by using an enzyme-linked immunosorbent assay (ELISA). Selection of various E
rns
fragments was based on an immunodominant E
rns
region encompassing three overlapping antigenic regions, amino acids 65 to 145 (E
rns
aa
65-145
) (AR1), 84 to 160 (E
rns
aa
84-160
) (AR2), and 109 to 220 (E
rns
aa
10
9-220
) (AR3), identified earlier by our group (M. Lin, E. Trottier, J. Pasick, and M. Sabara, J. Biochem., in press). Defined E
rns
fragments, including AR1, AR2, AR3, E
rns
aa
65-160
(AR12), E
rns
aa
84-220
(AR23), E
rns
aa
65-220
(AR123), E
rns
aa
109-145
(the consensus region defined by the three overlapping regions), and E
rns
aa
109-160
(a fragment 15 amino acids larger than the consensus region), were expressed in
Escherichia coli
, purified by nickel chelate affinity chromatography, and used to measure antibody responses in 20 sera serially collected from pigs experimentally infected with CSFV. Based on the optimum cutoffs determined by receiver operating characteristic analysis after testing 238 negative field sera from Canadian sources, all the E
rns
fragments were capable of distinguishing positive from negative antibody responses with sensitivities ranging between 75 and 90% and specificities ranging between 83.2 and 100%. Detection of antibody responses to refolded E
rns
aa
109-145
and E
rns
aa
109-160
by ELISA (this study) but not by Western blots (Lin et al., in press) indicated that the epitopes within the consensus region are conformational. When cutoff values were raised to give a specificity of 100%, four E
rns
fragments (AR2, AR23, E
rns
aa
109-145
, and E
rns
aa
109-160
) offered much higher sensitivities (75 to 90%) than those obtained with other fragments (20 to 65%). E
rns
aa
109-145
and E
rns
aa
109-160
were capable of detecting antibody responses in infected pigs as early as 7 days postinfection. Demonstration of antibody responses to either one of the four fragments can thus be an alternative to use of the full-length protein in ELISA for serological diagnosis of CSFV infection. An advantage of such a test would be its utilization for serological survey in a classical swine fever-free country (e.g., Canada) in biocontainment level 2 laboratories.
Expression of Recombinant Small Hydrophobic Protein for Serospecific Detection of Avian Pneumovirus Subgroup C
Authors: Luo Lizhong , Sabara Marta I. , Li Yan
Published: January 2005

The small hydrophobic (SH) gene of the avian pneumovirus (APV) Colorado isolate (CO), which belongs to subgroup C (APV/C), was expressed with a baculovirus vector. The recombinant SH protein was evaluated as a potential subgroup-specific diagnostic reagent in order to differentiate infections resulting from APV/C from those induced by APV/A, APV/B, and human metapneumovirus (hMPV). When the recombinant baculovirus was used to infect insect cells, a 31- to 38-kDa glycosylated form of the SH protein was produced and subsequently tested for reactivity with antibodies specific for APV/A, APV/B, APV/C, and hMPV. Western blot analysis showed that the expressed recombinant SH protein could only be recognized by APV/C-specific antibodies. This result was consistent with sequence analysis of the APV/C SH protein, which had very low (24%) amino acid identity with the corresponding protein of hMPV and no discernible identity with the SH protein of APV/A or APV/B. A recombinant SH protein-based enzyme-linked immunosorbent assay (ELISA) was developed, and it further confirmed the lack of reactivity of this protein with antisera raised to APV/A, APV/B, and hMPV and supported its designation as a subgroup-specific antigen. This finding indicated that the recombinant SH protein was a suitable antigen for ELISA-based detection of subgroup-specific antibodies in turkeys and could be used for serologically based differential diagnosis of APV and hMPV infections.
Visualizing Cytokine-Secreting Cells In Situ in the Rhesus Macaque Model of Chronic Gut Inflammation
Authors: Ramesh Geeta , Alvarez Xavier , Borda Juan T. , Aye Pyone P. , Lackner Andrew A. , Sestak Karol
Published: January 2005

Cytokine-producing cells in gut-associated lymphoid tissues of rhesus macaques with chronic enterocolitis were studied. The confocal microscopy technique that we developed enables simultaneous in situ visualization of multiple extra- and/or intracellular antigens at a resolution higher than that allowed by light or epifluorescence microscopy. The presence of interleukin-6 (IL-6)-, tumor necrosis factor alpha (TNF-α)-, and IL-1-α-producing cells was focally intense in the colon lamina propria of the affected animals. The IL-1-α-producing cells were T lymphocytes (CD3+), while the TNF-α-producing cells were both macrophages (CD68+/HAM56+/LN5+) and T lymphocytes (CD3+). The IL-6-producing cells within the colon consisted of T lymphocytes and macrophages. The amount of IL-6-producing cells seen in macaques with enterocolitis was significantly higher (P< 0.001) than that seen in the healthy control animal, while TNF-α- and IL-1-α-producing cells were seen only in macaques with enterocolitis. Most of the T lymphocytes that produced cytokines were detected in the lamina propria, while the macrophages were most prominent in highly inflamed regions of the lamina propria. Taken together, our findings indicate that there might be immunological similarity between chronic enterocolitis of rhesus macaques and humans, suggesting the potential use of the nonhuman primate model for the validation of novel therapies.
Fast Dipstick Dye Immunoassay for Detection of Immunoglobulin G (IgG) and IgM Antibodies of Human Toxoplasmosis
Authors: Jin Si , Chang Zhu Yin , Ming Xu , Min Cao Li , Wei He , Sheng Liang You , Hong Guan Xiao
Published: January 2005

A dipstick dye immunoassay (DDIA) was developed to detect immunoglobulin G (IgG) or IgM antibodies of toxoplasmosis infection in humans. The assays employ a blue colloidal dye particles (D-1) conjugated to sheep anti-human IgG and rabbit anti-human IgM as the visualizing agents and a soluble antigen of tachyzoites of
Toxoplasma gondii
strain RH (TSA) as the detective antigen. The mixture of dye-labeled anti-human antibody-special human antibody was captured by TSA onto a nitrocellulose membrane dipstick by means of immunochromatography. The assays are rapid (the whole test can be completed within 15 min), simple, and cheap, and they don't require any equipment. They are sensitive and specific for the detection of anti-
Toxoplasma
IgG or IgM antibodies and generally agree closely with the results from the enzyme-linked immunosorbent assay. The assays are especially suitable for field applications.
Susceptible and Protective Human Leukocyte Antigen Class II Alleles and Haplotypes in Bahraini Type 2 (Non-Insulin-Dependent) Diabetes Mellitus Patients
Authors: Motala Ayesha A. , Busson Marc , Al-Harbi Einas M. , Khuzam Manal A. A. , Al-Omari Emtiaz M. D. , Arekat Mona R. , Almawi Wassim Y.
Published: January 2005

Whereas the genetic risk for type 1 diabetes is linked to human leukocyte antigen (HLA) class II genes, the HLA association in type 2 (non-insulin-dependent) diabetes is less clear. The association between HLA class II genotypes and type 2 diabetes was examined in adult Bahrainis, an Arab population with a high prevalence of type 2 diabetes. HLA-DRB1* and -DQB1* genotyping of 86 unrelated type 2 diabetes patients (age, 51.6 ± 8.2 years; mean duration of diabetes, 7.7 ± 7.1 years) who had a strong family history of diabetes (52 of 72 versus 0 of 89 for controls,P< 0.001) and 89 healthy subjects was done by PCR-sequence-specific priming. DRB1*040101 (0.1221 versus 0.0562,P= 0.019) and DRB1*070101 (0.2151 versus 0.0843,P< 0.001) were positively associated, while DRB1*110101 (0.0698 versus 0.1461,P= 0.014) and DRB1*160101 (0.0640 versus 0.1236,P= 0.038) were negatively associated with type 2 diabetes. DRB1*040101-DQB1*0302 (0.069 versus 0.0007;P= 0.004), DRB1*070101-DQB1*0201 (0.178 versus 0.0761,P= 0.007), DRB1*070101-DQB1*050101 (0.125 versus 0.0310,P= 0.002), and DRB1*150101-DQB1*060101 (0.0756 versus 0.0281,P= 0.008) were more prevalent among patients, while DRB1*160101-DQB1*050101 (0.0702 versus 0.0349,P= 0.05) was more prevalent among controls, conferring disease susceptibility or protection, respectively. In Bahrainis with type 2 diabetes, there is a significant association with select HLA class II genotypes, which were distinct from those in type 1 diabetes.
Assignment of Weight-Based Immunoglobulin G1 (IgG1) and IgG2 Units in Antipneumococcal Reference Serum Lot 89-S(F) for Pneumococcal Polysaccharide Serotypes 1, 4, 5, 7F, 9V, and 18C
Authors: Sikkema Daniel J. , Ziembiec Nancy A. , Jones Thomas R. , Hildreth Stephen W. , Madore Dace V. , Quataert Sally A.
Published: January 2005

Weight-based assignments for immunoglobulin G1 (IgG1) and IgG2 subclass antibodies to
Streptococcus pneumoniae
capsular polysaccharides (PnPs) in antipneumococcal standard reference serum lot 89-S (lot 89-S), also known as lot 89-SF, have been determined for serotypes 1, 4, 5, 7F, 9V, and 18C. This extends the usefulness of lot 89-S beyond the IgG1 and IgG2 subclass assignments for serotypes 3, 6B, 14, 19F, and 23F made previously (A. Soininen, H. Kayhty, I. Seppala, and T. Wuorimaa, Clin. Diagn. Lab. Immunol.
5:
561-566, 1998) to cover 11 major serotypes associated with the highest percentage of pneumococcal disease worldwide. A method of equivalence of absorbances in enzyme immunosorbent assays was used to determine the IgG1 and IgG2 antibody concentrations for the additional serotypes in lot 89-S, based on the subclass values previously assigned for PnPs serotypes 6B, 14, and 23F. This cross-standardization method assures consistency with previous antibody assignments in that reference serum. The newly assigned subclass values for serotype 9V, and previously assigned values for serotype 14, were used to quantitate PnPs antibodies in sera from adult and pediatric subjects immunized with a pneumococcal conjugate vaccine. There was a predominance of IgG1 anti-PnPs antibodies in pediatric sera and IgG2 anti-PnPs antibodies in the adult sera. The IgG1 and IgG2 subclass assignments for the 11 PnPs serotypes in antipneumococcal standard reference serum lot 89-S are useful for quantitating and characterizing immune responses to pneumococcal infection and vaccination regimens.
Comparison of a New, Affordable Flow Cytometric Method and the Manual Magnetic Bead Technique for CD4 T-Lymphocyte Counting in a Northern Nigerian Setting
Authors: Imade Godwin E. , Badung Bitrus , Pam Sunday , Agbaji Oche , Egah Daniel , Sagay Atiene S. , Sankalé Jean-Louis , Kapiga Saidi , Idoko John , Kanki Phyllis
Published: January 2005

We compared two techniques for CD4 T-lymphocyte counting: flow cytometry (Cyflow) and magnetic beads (Dynabead). Similar results with good correlation were obtained from the 40 adult blood samples counted (
P
= 0.057,
r
= 0.93). The Cyflow technique is more precise and cost-effective than the Dynabead method ($3 to $5 versus $12 to $22 per test, respectively), since as many as 200 samples can be measured per day.
Effects of Lymphocyte Isolation and Timing of Processing on Detection of CD127 Expression on T Cells in Human Immunodeficiency Virus-Infected Patients
Authors: Higgins Jeanette , Metcalf Julia A. , Stevens Randy A. , Baseler Michael , Nason Martha C. , Lane H. Clifford , Sereti Irini
Published: January 2005

Decreases in the detection of CD127 expression on T cells of human immunodeficiency virus-infected patients by flow cytometry can occur by delayed processing or by peripheral blood mononuclear cell isolation and cryopreservation. These observations should be considered in the interpretation of functional studies and the planning of multicenter clinical trials.
Prevalence of Antibodies to
Onchocerca volvulus
in Residents of Oaxaca, Mexico, Treated for 10 Years with Ivermectin
Authors: Gómez-Priego Alberto , Mendoza Raymundo , de-la-Rosa Jorge-Luis
Published: January 2005

Studies to determine the prevalence of antibodies to
Onchocerca volvulus
, prior to and after actions carried out to interrupt transmission, are scarce in Mexico. Here we report the prevalence of immunoglobulin G (IgG) and IgG4 antibodies in an enzyme-linked immunosorbent assay (ELISA) against a crude extract of
O. volvulus
adult worm in serum samples from persons under noninterrupted biannual treatment with ivermectin in areas of onchocercosis endemicity in Mexico. To perform the prevalence studies, the ELISA procedures were first evaluated. Serological studies were performed with serum samples from skin microfilaria carriers from Guatemala and from people microfilariodermic negative living in the same area as the Guatemalan patients. Sensitivity values for IgG or IgG4 detection were 71 and 86%, while specificities were 92 and 100%, respectively. No anti-
O. volvulus
antibodies were found in samples from nonendemic controls from Mexico, but 3 of 71 samples from residents in the onchocercosis area of Oaxaca, Mexico, and who have been under ivermectin treatment during the last 10 years were only positive to IgG. Notwithstanding that the IgG4 isotype was not detected and a low (4.2%) anti-
O. volvulus
IgG antibody prevalence was found, a seroepidemiological follow-up must be performed in order to confirm interruption of onchocercosis transmission in the area of Oaxaca, Mexico, in which onchocercosis is endemic.
Spinal Fluid Abnormalities in Patients with Chronic Fatigue Syndrome
Authors: Natelson Benjamin H. , Weaver Shelley A. , Tseng Chin-Lin , Ottenweller John E.
Published: January 2005

Arguments exist as to the cause of chronic fatigue syndrome (CFS). Some think that it is an example of symptom amplification indicative of functional or psychogenic illness, while our group thinks that some CFS patients may have brain dysfunction. To further pursue our encephalopathy hypothesis, we did spinal taps on 31 women and 13 men fulfilling the 1994 case definition for CFS and on 8 women and 5 men serving as healthy controls. Our outcome measures were white blood cell count, protein concentration in spinal fluid, and cytokines detectable in spinal fluid. We found that significantly more CFS patients had elevations in either protein levels or number of cells than healthy controls (30 versus 0%), and 13 CFS patients had protein levels and cell numbers that were higher than laboratory norms; patients with abnormal fluid had a lower rate of having comorbid depression than those with normal fluid. In addition, of the 11 cytokines detectable in spinal fluid, (i) levels of granulocyte-macrophage colony-stimulating factor were lower in patients than controls, (ii) levels of interleukin-8 (IL-8) were higher in patients with sudden, influenza-like onset than in patients with gradual onset or in controls, and (iii) IL-10 levels were higher in the patients with abnormal spinal fluids than in those with normal fluid or controls. The results support two hypotheses: that some CFS patients have a neurological abnormality that may contribute to the clinical picture of the illness and that immune dysregulation within the central nervous system may be involved in this process.
Acute Inflammatory Response to Endotoxin in Mice and Humans
Authors: Copeland Shannon , Warren H. Shaw , Lowry Stephen F. , Calvano Steve E. , Remick Daniel
Published: January 2005

Endotoxin injection has been widely used to study the acute inflammatory response. In this study, we directly compared the inflammatory responses to endotoxin in mice and humans.Escherichia colitype O113 endotoxin was prepared under identical conditions, verified to be of equal biological potency, and used for both mice and humans. The dose of endotoxin needed to induce an interleukin-6 (IL-6) concentration in plasma of ∼1,000 pg/ml 2 h after injection was 2 ng/kg of body weight in humans and 500 ng/kg in mice. Healthy adult volunteers were injected intravenously with endotoxin, and male C57BL/6 mice (n= 4 to 12) were injected intraperitoneally with endotoxin. Physiological, hematological, and cytokine responses were determined. Endotoxin induced a rapid physiological response in humans (fever, tachycardia, and slight hypotension) but not in mice. Both mice and humans exhibited lymphopenia with a nadir at 4 h and recovery by 24 h. The levels of tumor necrosis factor (TNF) and IL-6 in plasma peaked at 2 h and returned to baseline levels by 4 to 6 h. IL-1 receptor antagonist RA and TNF soluble receptor I were upregulated in both mice and humans but were upregulated more strongly in humans. Mice produced greater levels of CXC chemokines, and both mice and humans exhibited peak production at 2 h. These studies demonstrate that although differences exist and a higher endotoxin challenge is necessary in mice, there are several similarities in the inflammatory response to endotoxin in mice and humans.