bluuebunny/crossref_metadata_2025 huggingface dataset:
Veterans Health Administration and Medicare Outpatient Health Care Utilization by Older Rural and Urban New England Veterans
Authors: Weeks William B. , Bott David M. , Lamkin; Rebecca P. , Wright Steven M.
Published: June 2006

ABSTRACT: Context: Older veterans often use both the Veterans Health Administration (VHA) and Medicare to obtain health care services. Purpose: The authors sought to compare outpatient medical service utilization of Medicare‐enrolled rural veterans with their urban counterparts in New England. Methods: The authors combined VHA and Medicare databases and identified veterans who were age 65 and older and enrolled in Medicare fee‐for‐service plans, and they obtained records of all their VHA services in New England between 1997 and 1999. The authors used ZIP codes to designate rural or urban residence and categorized outpatient utilization into primary care, individual mental health care, non–mental health specialty care, or emergency room care. Findings: Compared with their urban counterparts, veterans living in rural settings used significantly fewer VHA and Medicare‐funded primary care, specialist care, and mental health care visits in all 3 years examined (P<.001 for all). Compared with urban veterans, veterans living in rural settings used fewer VHA emergency department services in 1998 and 1999 but more Medicare‐funded emergency department visits in 1997. The authors found some evidence of substitution of Medicare for VHA emergency visits in rural veterans, but no other evidence of like‐service substitution. Rural veterans were more reliant on Medicare for primary care and on VHA services for specialty and mental health care. Conclusions: These findings suggest that rural access to federally funded health care is restricted relative to urban access. Older veterans may choose different systems of care for different health care services. With poor access to primary care, rural veterans may substitute emergency room visits for routine care.
Improvements in Care and Reduced Self‐management Barriers Among Rural Patients With Diabetes
Authors: Dettori Nancy , Flook Benjamin N. , Pessl Erich , Quesenberry Kim , Loh Johnson , Harris Colleen , McDowall Janet M. , Butcher Marcene K. , Helgerson Steven D. , Gohdes Dorothy , Harwell Todd S.
Published: June 2006

ABSTRACT: Context: Improved preventive care and clinical outcomes among patients with diabetes can reduce complications and costs; however, diabetes care continues to be suboptimal. Few studies have described effective strategies for improving care among rural populations with diabetes. Purpose: In 2000, the Park County Diabetes Project and the Montana Diabetes Control Program collaboratively implemented a countywide effort, which included health systems interventions and coordinated diabetes education, to improve the quality of diabetes care. Methods: Clinical data from the diabetes registries in 2 primary care practices, in addition to baseline and follow‐up telephone surveys, were used to evaluate improvements in care, outcomes, education, and barriers to self‐management. Findings: In the cohort of patients, the proportion receiving the following services increased significantly from 2000 to 2003: annual foot examination (43% to 58%), influenza (30% to 53%), and pneumoccocal immunizations (39% to 70%). The median hemoglobin A1c values decreased significantly from baseline to follow‐up (7.2% to 6.8%). Mean systolic and diastolic blood pressure decreased significantly over the 2 time periods (139 mmHg to 135 mmHg, and 78 mmHg to 75 mmHg, respectively). Significant decreases were also observed in barriers to self‐management, including lack of knowledge (decrease from 12% to 5%), difficulties making lifestyle changes (36% to 27%), cost of monitors and test strips (25% to 16%), cost of medications (37% to 24%), and diabetes education (22% to 4%). Conclusions: Findings suggest that system changes in primary care practices and the implementation of accessible diabetes education can improve care and reduce barriers for rural patients with diabetes.
Gender differences in the insulin‐like growth factor axis response to a glucose load
Authors: Flanagan D. E. , Holt R. I. G. , Owens P. C. , Cockington R. J. , Moore V. M. , Robinson J. S. , Godsland I. F. , Phillips D. I. W.
Published: June 2006

Aims:  The insulin‐like growth factors (IGFs) are thought to contribute to glucose homeostasis. The aim of our study was to examine the response of the IGFs and their binding proteins to an intravenous load of glucose in a cohort of young men and women with normal glucose tolerance.

Methods:  The intravenous glucose tolerance test (IVGTT) was used to quantify insulin sensitivity and insulin secretion in 160 adults aged 20–21 years in Adelaide, Australia. Serum IGF‐I, IGF‐II, IGF‐binding protein (IGFBP)‐1 and IGFBP‐3 were measured during the IVGTT.

Results:  Women were less insulin sensitive than men with higher fasting insulin (women 55.6 ± 4.4, men 44.1 ± 3.6 pmol L−1, P = 0.001) and first phase insulin secretion (women 3490 ± 286, men 3038 ± 271 pmol L−1 min, P = 0.042). Women showed lower fasting free IGF‐I (women 0.29 ± 0.02, men 0.36 ± 0.02 μg L−1, P = 0.004) but higher IGFBP‐3 (women 46.3 ± 0.53, men 43.3 ± 0.58 mg dL−1, P = 0.001) and higher IGFBP‐1 concentrations (women 37.0 ± 2.9, men 24.8 ± 2.3 μg L−1, P = 0.012). IGFBP‐1 fell by 5 min and remained suppressed. IGFBP‐3 and total IGF‐I fell until 60 min rising again by 2 h. IGF and IGFBP values were all higher in women. IGFBP‐1 showed a negative association with fasting and stimulated insulin concentrations in both genders. First phase insulin secretion however showed positive correlations with IGFBP‐3 (r = 0.321, P = 0.004) and IGF‐I (r = 0.339 P = 0.002) in men but not women.

Conclusion:  Our data show that IGFBP‐1, IGFBP‐3 and IGF‐I show acute changes following a glucose load and there are marked gender differences in these responses.
Correlation of Cytokine Gene Expression with Pathology in White-Tailed Deer (
Odocoileus virginianus
) Infected with
Mycobacterium bovis
Authors: Thacker Tyler C. , Palmer Mitchell V. , Waters W. Ray
Published: June 2006

Mycobacterium bovis
-infected white-tailed deer (WTD) in northeast Michigan are a reservoir of mycobacteria that pose a threat to both domestic animals and humans. Relatively little work has been done to characterize the immune response of WTD to
M. bovis
infection; however, an understanding of the immune response to infection and pathogenesis may be critical to the development of an effective vaccine. Immunological responses to infection were characterized by monitoring cytokine gene expression in
M. bovis
-infected and uninfected deer. Peripheral blood leukocytes (PBL) from infected WTD expressed more gamma interferon (IFN-γ), interleukin-12p40 (IL-12p40), granulocyte-monocyte colony-stimulating factor, and IL-4 mRNA than did PBL from uninfected deer; however, differences were not detected in expression of IL-10 and transforming growth factor-β mRNA. Infected animals could be divided into two groups based on pathology. Lesions were confined primarily to the lymph nodes of the head in animals with less severe pathology. Animals with more severe pathology had lesions in the lung and associated lymph nodes as well as the lymph nodes of the head. More robust IFN-γ mRNA expression correlated with pathology early in infection. These findings indicate that IFN-γ expression likely plays a role in both protection and pathogenesis.
Immune Responses to Defined Antigens of
Mycobacterium bovis
in Cattle Experimentally Infected with
Mycobacterium kansasii
Authors: Waters W. R. , Palmer M. V. , Thacker T. C. , Payeur J. B. , Harris N. B. , Minion F. C. , Greenwald R. , Esfandiari J. , Andersen P. , McNair J. , Pollock J. M. , Lyashchenko K. P.
Published: June 2006

Cross-reactive responses elicited by exposure to nontuberculous mycobacteria often confound the interpretation of antemortem tests for
Mycobacterium bovis
infection of cattle. The use of specific proteins (e.g., ESAT-6, CFP-10, and MPB83), however, generally enhances the specificity of bovine tuberculosis tests. While genes for these proteins are absent from many nontuberculous mycobacteria, they are present in
M. kansasii
. Instillation of
M. kansasii
into the tonsillar crypts of calves elicited delayed-type hypersensitivity and in vitro gamma interferon and nitrite concentration responses of leukocytes to
M. avium
and
M. bovis
purified protein derivatives (PPDs). While the responses of
M. kansasii
-inoculated calves to
M. avium
and
M. bovis
PPDs were approximately equivalent, the responses of
M. bovis
-inoculated calves to
M. bovis
PPD exceeded their respective responses to
M. avium
PPD. The gamma interferon and nitrite responses of
M. kansasii
-inoculated calves to recombinant ESAT-6-CFP-10 (rESAT-6-CFP-10) exceeded corresponding responses of noninoculated calves as early as 15 and 30 days after inoculation, respectively, and persisted throughout the study. The gamma interferon and nitrite responses of
M. bovis
-inoculated calves to rESAT-6-CFP-10 exceeded the corresponding responses of
M. kansasii
-inoculated calves beginning 30 days after inoculation. By using a lipoarabinomannan-based enzyme-linked immunosorbent assay, specific serum antibodies were detected as early as 50 days after challenge with
M. kansasii
. By a multiantigen print immunoassay and immunoblotting, serum antibodies to MPB83, but not ESAT-6 or CFP-10, were detected in
M. kansasii
-inoculated calves; however, responses to MPB83 were notably weaker than those elicited by
M. bovis
infection. These findings indicate that
M. kansasii
infection of calves elicits specific responses that may confound the interpretation of bovine tuberculosis tests.
Comparison of Two Hepatitis B Vaccines (GeneVac-B and Engerix-B) in Healthy Infants in India
Authors: Shivananda  , Somani Virbhadra , Srikanth B. S. , Mohan M. , Kulkarni P. S.
Published: June 2006

Hepatitis B is a major problem in many parts of the world. The WHO has recommended the inclusion of hepatitis B vaccines in routine immunization schedules. We wanted to compare two recombinant hepatitis B vaccines in an infant population for immunogenicity and reactogenicity when given at 6, 10, and 14 weeks of age. One hundred seventy-three infants meeting eligibility criteria were given either GeneVac-B (Serum Institute of India Ltd.) or Engerix-B (GlaxoSmithKline Beecham) in a random fashion. Three 0.5-ml (10-μg) doses of the vaccines were given at 6, 10, and 14 weeks of age along with diphtheria-pertussis (whole cell)-tetanus (DTPw) vaccine. Blood samples were collected at baseline and 1 month after administration of the third dose of the vaccines to measure anti-HBs antibody levels. Seroconversion was defined as a titer of more than 1 × 10
−3
IU/ml, while seroprotection was defined as a titer of more than 10 × 10
−3
IU/ml. Of the GeneVac-B recipients, 98% seroconverted versus 99% of the Engerix-B group. The anti-HBs geometric mean titer was slightly greater for GeneVac-B (229 × 10
−3
IU/ml) than for Engerix-B (167 × 10
−3
IU/ml), but the difference was not significant. The seroprotection rates were similar for both vaccines (96% and 95%, respectively). The most common systemic reaction events were mild to moderate fever, excessive crying, local swelling, rash, and irritability, and the local reactions were redness, induration, and edema, which most probably were caused by the simultaneously administered DTPw vaccine. All events were transient and resolved without sequelae. Reactogenicity was similar for the two vaccines. The present study shows that GeneVac-B is as immunogenic and as well tolerated as Engerix-B when administered with DTPw vaccine at 6, 10, and 14 weeks of age.
Sequence-Based Typing of
Mycoplasma genitalium
Reveals Sexual Transmission
Authors: Hjorth Sofie Vetli , Björnelius Eva , Lidbrink Peter , Falk Lars , Dohn Birthe , Berthelsen Lene , Ma Liang , Martin David H. , Jensen Jørgen Skov
Published: June 2006

Mycoplasma genitalium
causes male nonchlamydial, nongonococcal urethritis and is associated with cervicitis and pelvic inflammatory disease in women. Epidemiological studies indicate that
M. genitalium
is sexually transmitted, and the aim of the present study was to further substantiate this by means of a DNA typing system. A typing assay based on a diagnostic
mgpB
gene PCR was developed, evaluated, and applied directly to urogenital specimens. The assay had a low limit of detection and hence a high typeability. Sequences of isolates from 52 unrelated patients were divided into 29 different sequence types, giving a discriminatory index of 0.95. Two to six
M. genitalium
-positive specimens were collected from each of 44 patients over a median interval of 56 days (range, 11 to 1,395). Forty had the same sequence type in consecutive specimens. Specimens collected from two men were repeatedly positive at intervals of 472 and 1,395 days, respectively, but the sequence types had changed. A new strain was introduced in one sexual dyad, and the sequence types changed subsequently. Seventy-nine
M. genitalium
-positive specimens from 19 couples were investigated, and all partners initially had concordant sequence types, but one couple had discordant types at one time point before a newly introduced strain took over. The present typing system is simple and reproducible and has an excellent discriminatory capacity which might prove useful in studies of sexual networks and for evaluation of treatment failures. In the laboratory, this system may document the uniqueness of newly isolated
M. genitalium
strains.
Enhanced Detection and Typing of Human Papillomavirus (HPV) DNA in Anogenital Samples with PGMY Primers and the Linear Array HPV Genotyping Test
Authors: Coutlée François , Rouleau Danielle , Petignat Patrick , Ghattas Georges , Kornegay Janet R. , Schlag Peter , Boyle Sean , Hankins Catherine , Vézina Sylvie , Coté Pierre , Macleod John , Voyer Hélène , Forest Pierre , Walmsley Sharon ,   The Canadian Women's HIV Study Group, Franco Eduardo
Published: June 2006

The Roche PGMY primer-based research prototype line blot assay (PGMY-LB) is a convenient tool in epidemiological studies for the detection and typing of human papillomavirus (HPV) DNA. This assay has been optimized and is being commercialized as the Linear Array HPV genotyping test (LA-HPV). We assessed the agreement between LA-HPV and PGMY-LB for detection and typing of 37 HPV genotypes in 528 anogenital samples (236 anal, 146 physician-collected cervical, and 146 self-collected cervicovaginal swabs) obtained from human immunodeficiency virus-seropositive individuals (236 men and 146 women). HPV DNA was detected in 433 (82.0%) and 458 (86.7%) samples with PGMY-LB and LA-HPV (P= 0.047), respectively, for an excellent agreement of 93.8% (kappa = 0.76). Of the 17,094 HPV typing results, 16,562 (1,743 positive and 14,819 negative results) were concordant between tests (agreement = 96.9%; kappa = 0.76). The mean agreement between tests for each type was 96.4% ± 2.4% (95% confidence interval [CI], 95.6% to 97.2%; range, 86% to 100%), for an excellent mean kappa value of 0.85 ± 0.10 (95% CI, 0.82 to 0.87). However, detection rates for most HPV types were greater with LA-HPV. The mean number of types per sample detected by LA-HPV (4.2 ± 3.4; 95% CI, 3.9 to 4.5; median, 3.0) was greater than that for PGMY-LB (3.4 ± 3.0; 95% CI, 3.1 to 3.6; median, 2.0) (P< 0.001). The number of types detected in excess by LA-HPV in anal samples correlated with the number of types per sample (r= 0.49 ± 0.06;P= 0.001) but not with patient age (r= 0.03 ± 0.06;P= 0.57), CD4 cell counts (r= 0.06 ± 0.06;P= 0.13), or the grade of anal disease (r= −0.11 ± 0.06;P= 0.07). LA-HPV compared favorably with PGMY-LB but yielded higher detection rates for newer and well-known HPV types.
Value of the Chlorhexidine Decontamination Method for Recovery of Nontuberculous Mycobacteria from Sputum Samples of Patients with Cystic Fibrosis
Authors: Ferroni Agnès , Vu-Thien Hoang , Lanotte Philippe , Le Bourgeois Muriel , Sermet-Gaudelus Isabelle , Fauroux Brigitte , Marchand Sophie , Varaigne Françoise , Berche Patrick , Gaillard Jean-Louis , Offredo Catherine
Published: June 2006

The chlorhexidine method was compared to the
N
-acetyl-
l
-cysteine-NaOH-oxalic acid decontamination method currently recommended for the recovery of nontuberculous mycobacteria (NTM) from patients with cystic fibrosis. Sputum samples (
n
= 827) treated with chlorhexidine yielded twice as many NTM-positive cultures as those treated by the reference method (54 [6.50%] versus 27 [3.25%];
P
< 0.0001) despite a higher contamination rate (20% versus 14.2%;
P
= 0.0017).
Rapid Discrimination of
Listeria monocytogenes
Strains by Microtemperature Gradient Gel Electrophoresis
Authors: Tominaga Tatsuya
Published: June 2006

Microtemperature gradient gel electrophoresis (μ-TGGE) was examined for use for the rapid subtyping of
Listeria monocytogenes
strains. Comparison of genomes between
L. monocytogenes
strains F2365 and H7858 identified a sequence encoding a portion of the PRT/PTS system IIA 2 protein domain as appropriate for μ-TGGE analysis. Thirty-one strains belonging to 10 different serovar types were tested by PCR, and sequence analysis of the amplified products revealed that the strains comprise 11 groups. All 55 possible pairs within the 11 groups were examined by μ-TGGE analysis. Of these, 47 pairs could be successfully discriminated, with a total electrophoresis time of only 7 min. Moreover, Cy3/Cy5 labeling allowed rapid identification of the sequence type in unknown strains of
L. monocytogenes
isolated from meat. These findings collectively indicate that μ-TGGE can be used for the rapid analysis of
L. monocytogenes
strains, facilitating determination of routes of contamination when these bacteria are found in food products.
Identification and Characterization of Variable-Number Tandem-Repeat Markers for Typing of
Brucella
spp
Authors: Whatmore Adrian M. , Shankster Stephen J. , Perrett Lorraine L. , Murphy Terry J. , Brew Simon D. , Thirlwall Rachel E. , Cutler Sally J. , MacMillan Alastair P.
Published: June 2006

Members of the genus
Brucella
infect many domesticated and wild animals and cause serious zoonotic infection in humans. The availability of discriminatory molecular typing tools to inform and assist conventional epidemiological approaches would be invaluable in controlling these infections, but efforts have been hampered by the genetic homogeneity of the genus. We report here on a molecular subtyping system based on 21 variable-number tandem-repeat (VNTR) loci consisting of 13 previously unreported loci and 8 loci previously reported elsewhere. This approach was applied to a collection of 121
Brucella
isolates obtained worldwide and representing all six classically recognized
Brucella
species. The size of repeats selected for inclusion varied from 5 to 40 bp giving VNTR loci with a range of diversities. The number of alleles detected ranged from 2 to 21, and Simpson's diversity index values ranged from 0.31 to 0.92. This assay divides the 121 isolates into 119 genotypes, and clustering analysis results in groups that, with minor exceptions, correspond to conventional species designations. Reflecting this, the use of six loci in isolation was shown to be sufficient to determine species designation. On the basis of the more variable loci, the assay could also discriminate isolates originating from restricted geographical sources, indicating its potential as an epidemiological tool. Stability studies carried out in vivo and in vitro showed that VNTR profiles were sufficiently stable such that recovered strains could readily be identified as the input strain. The method described here shows great potential for further development and application to both epidemiological tracing of
Brucella
transmissions and in determining relationships between isolates worldwide.
Staphylococcus aureus
Carriage Patterns and the Risk of Infections Associated with Continuous Peritoneal Dialysis
Authors: Nouwen Jan , Schouten Jeroen , Schneebergen Peter , Snijders Susan , Maaskant Jolanda , Koolen Marjan , van Belkum Alex , Verbrugh Henri A.
Published: June 2006

The epidemiology and risks of
Staphylococcus aureus
carriage in continuous peritoneal dialysis (CPD) patients was studied in a single tertiary-care institution. On outpatient visits samples for culture were routinely taken prospectively from the CPD catheter exit site and the vestibulum nasi. Seventy-five patients with at least one culture positive for
S. aureus
in this period were included: 43 had genotypically identical
S. aureus
strains in over 80% of the cultures and were classified as persistent carriers; 32 were intermittent carriers. Persistent carriage was associated with a threefold higher risk for CPD-related infections and sixfold higher rates of vancomycin consumption compared to those for the intermittent carriers. No methicillin or vancomycin resistance was detected.
Rapid Detection of Rifampin Resistance in
Mycobacterium tuberculosis
by Pyrosequencing Technology
Authors: Jureen Pontus , Engstrand Lars , Eriksson Solveig , Alderborn Anders , Krabbe Margareta , Hoffner Sven E.
Published: June 2006

We developed an assay for rapid detection of rifampin resistance in
Mycobacterium tuberculosis
based on Pyrosequencing technology, involving a technique for real-time sequencing. A 180-bp region of the
rpoB
gene was amplified in clinical isolates of both rifampin-resistant and -susceptible
M. tuberculosis
. The PCR products were subjected to Pyrosequencing analysis using four different sequencing primers in four overlapping reactions. These four sequencing reactions covered the 81-bp region where >96% of the mutations associated with rifampin resistance are located. The results were compared to those obtained with two other molecular methods, the line probe assay and cycle sequencing, and the phenotypic BACTEC method. The genotypic determination methods all detected the mutations that previously have been correlated with rifampin resistance. In addition, Pyrosequencing analysis and the two other molecular methods found additional mutations within the
rpoB
gene in phenotypically susceptible strains. We found that Pyrosequencing technology, in particular, offers high accuracy, short turnaround time, and a potentially high throughput in detection of rifampin resistance in
M. tuberculosis
.
Combining Multiplex Reverse Transcription-PCR and a Diagnostic Microarray To Detect and Differentiate Enterovirus 71 and Coxsackievirus A16
Authors: Chen Tsan-Chi , Chen Guang-Wu , Hsiung Chao Agnes , Yang Jyh-Yuan , Shih Shin-Ru , Lai Yiu-Kay , Juang Jyh-Lyh
Published: June 2006

Cluster A enteroviruses, including enterovirus 71 (EV71) and coxsackievirus A16 (CA16), are known to cause hand-foot-and-mouth disease (HFMD). Despite the close genetic relationship between these two viruses, EV71 is generally known to be a more perpetuating pathogen involved in severe clinical manifestations and deaths. While the serotyping of enteroviruses is mostly done by conventional immunological methods, many clinical isolates remain unclassifiable due to the limited number of antibodies against enterovirus surface proteins. Array-based assays are able to detect several serotypes with high accuracy. We combined an enterovirus microarray with multiplex reverse transcription-PCR to try to develop a method of sensitively and accurately detecting and differentiating EV71 and CA16. In an effort to design serotype-specific probes for detection of the virus, we first did an elaborate bioinformatic analysis of the sequence database derived from different enterovirus serotypes. We then constructed a microarray using 60-mer degenerate oligonucleotide probes covalently bound to array slides. Using this enterovirus microarray to study 144 clinical specimens from patients infected with HFMD or suspected to have HFMD, we found that it had a diagnostic accuracy of 92.0% for EV71 and 95.8% for CA16. Diagnostic accuracy for other enteroviruses (non-EV71 or -CA16) was 92.0%. All specimens were analyzed in parallel by real-time PCR and subsequently confirmed by neutralization tests. This highly sensitive array-based assay may become a useful alternative in clinical diagnostics of EV71 and CA16.
Evaluation of a Multiple-Cycle, Recombinant Virus, Growth Competition Assay That Uses Flow Cytometry To Measure Replication Efficiency of Human Immunodeficiency Virus Type 1 in Cell Culture
Authors: Dykes Carrie , Wang Jiong , Jin Xia , Planelles Vicente , An Dong Sung , Tallo Amanda , Huang Yangxin , Wu Hulin , Demeter Lisa M.
Published: June 2006

Human immunodeficiency virus type 1 (HIV-1) replication efficiency or fitness, as measured in cell culture, has been postulated to correlate with clinical outcome of HIV infection, although this is still controversial. One limitation is the lack of high-throughput assays that can measure replication efficiency over multiple rounds of replication. We have developed a multiple-cycle growth competition assay to measure HIV-1 replication efficiency that uses flow cytometry to determine the relative proportions of test and reference viruses, each of which expresses a different reporter gene in place of
nef
. The reporter genes are expressed on the surface of infected cells and are detected by commercially available fluorescence-labeled antibodies. This method is less labor-intensive than those that require isolation and amplification of nucleic acids. The two reporter gene products are detected with similar specificity and sensitivity, and the proportion of infected cells in culture correlates with the amount of viral p24 antigen produced in the culture supernatant. HIV replication efficiencies of six different drug-resistant site-directed mutants were reproducibly quantified and were similar to those obtained with a growth competition assay in which the relative proportion of each variant was measured by sequence analysis, indicating that recombination between the
pol
and reporter genes was negligible. This assay also reproducibly quantified the relative fitness conferred by protease and reverse transcriptase sequences containing multiple drug resistance mutations, amplified from patient plasma. This flow cytometry-based growth competition assay offers advantages over current assays for HIV replication efficiency and should prove useful for the evaluation of patient samples in clinical trials.
Molecular Epidemiology of Group A Rotavirus Diarrhea among Children in Buenos Aires, Argentina, from 1999 to 2003 and Emergence of the Infrequent Genotype G12
Authors: Castello A. A. , Argüelles M. H. , Rota R. P. , Olthoff A. , Jiang B. , Glass R. I. , Gentsch J. R. , Glikmann G.
Published: June 2006

To examine the epidemiology of rotaviruses in Buenos Aires, Argentina, we screened 1,212 stool samples from children with diarrhea in the southern district of Buenos Aires from 1999 to 2003. We identified 187 samples (15.4%) that were positive for group A rotavirus by use of antigen enzyme-linked immunosorbent assay. Among these specimens, 112 were available for typing: 93 (83.0%) were single-type infections, 9 (8.0%) were mixed-type infections with more than one G or P type, and 10 (8.9%) were G and/or P nontypeable. In contrast to the findings in our last study, from 1996 to 1998, genotype P[4], G2 strains were almost completely absent and P[8], G1 and P[8], G4 strains were dominant, representing more than 80% of the G and P types found. Genotypes G2 and G9 were detected in few samples, and type G3 was completely absent. We identified several uncommon genotype G12 strains, representing the first detections outside of Asia and the United States, by sequencing. Using a genotype G12-specific reverse transcription-PCR, we identified eight (6.7%) positive samples for the 1999 to 2003 period. The high degree of sequence identity between recent G12 isolates from Argentina, the United States, and Asian countries suggests a relatively recent introduction(s) of these strains into humans from a common progenitor. The Argentinean G12 strains belonged to genotype P[9], similar to most of the recently described Asian G12 strains. The finding of G12 strains in several other regions of the world raises the possibility that G12 may be emerging globally and suggests that surveillance for this strain should be conducted routinely.
Genetically Diverse Group A Streptococci from Children in Far-Western Nepal Share High Genetic Relatedness with Isolates from Other Countries
Authors: Sakota Varja , Fry Alicia M. , Lietman Thomas M. , Facklam Richard R. , Li Zhongya , Beall Bernard
Published: June 2006

The genetic diversity of group A streptococci (GAS) throughout much of the world has not been adequately explored. To assess genetic variation among GAS in western Nepal, 120 noninvasive GAS, collected from eight different villages, were genetically characterized using
emm
typing,
sof
sequencing, and multilocus sequence typing (MLST). A high level of genetic diversity was observed among these isolates, with 51 genotypes based upon 51 multilocus sequence types (STs), 45
emm
sequence types, and 28
sof
sequence types. On the basis of shared ST
-emm
and
sof
-
emm
associations, 40 of the 51 genotypes were identical or highly related to genotypes characterized from locations outside of Nepal, even though most of the
emm
sequence and clonal types are rare among GAS within the United States. When analyzing all known STs highly related to Nepal STs, only one example of similar STs shared between a
sof
PCR-positive strain and a
sof
PCR-negative strain was observed. Since previous data indicate free exchange of MLST loci between
sof
-positive and
sof
-negative strains, there is possibly selection against the expansion of subclones resulting from horizontal transfers of
sof
or
emm
genes between
sof
-positive and
sof
-negative strains. All 45
emm
types encountered in Nepal have also been documented from other countries. These data, together with data encompassing the past decade of
emm
type surveillance, support the possibility that most existing GAS
emm
types have been discovered. Similarly, since most (40/51) strain types were highly related to strains found elsewhere, it is likely that a major fraction of the existing GAS clonal complexes have been discovered.
Analysis of the 5′ Noncoding Region versus the NS5b Region in Genotyping Hepatitis C Virus Isolates from Blood Donors in France
Authors: Cantaloube Jean-François , Laperche Syria , Gallian Pierre , Bouchardeau Françoise , de Lamballerie Xavier , de Micco Philippe
Published: June 2006

The 5′ noncoding region (5′ NCR) of the hepatitis C virus (HCV) has become the standard for genotyping even though several reports show that its use can result in classification errors. The purpose of this study was to perform genotyping based on sequence analysis of the NS5b region in a set of 357 HCV strains isolated from blood donors in France in 2002 and 2003. Results were compared with those previously obtained using 5′ NCR analysis, and HCV subtype distribution was reevaluated. Twenty-six of 120 strains (∼22%) initially identified as genotype 1b by 5′ NCR region sequence analysis were reclassified as genotype 1a by NS5b region sequence analysis. Similarly, 14 of 23 strains (∼61%) initially identified as 2a/2c were reclassified as non-2a and non-2c subtypes, and 12 of 22 strains (∼45%) initially identified as 4c/4d subtypes were reclassified as non-4c and non-4d subtypes. Sequence analysis of the NS5b region also revealed 5 putative new subtype 2 variants and 2 putative new subtype 4 variants. Although these findings demonstrated full agreement between 5′ NCR and NS5b sequence analysis with regard to type classification, genotyping based on phylogenetic analysis of the NS5b region is more accurate for subtype determination than genotyping based on analysis of the 5′ NCR. Sequence analysis of the NS5b region is mandatory for epidemiologic studies.
Acute Cholecystitis Accompanied by Acute Pancreatitis Potentially Caused by
Dolosigranulum pigrum
Authors: Lin Jung-Chung , Hou Shu-Jin , Huang Li-Ung , Sun Jun-Ren , Chang Wei-Kuo , Lu Jang-Jih
Published: June 2006

We report a case of acute cholecystitis accompanied by acute pancreatitis and caused by
Dolosigranulum pigrum
in a 76-year-old male with gallstones.
D. pigrum
was isolated from a blood culture and confirmed by biochemistry tests and 16S rRNA sequencing. The isolate was susceptible to the β-lactams ampicillin, penicillin, cephalothin, ceftriaxone, ceftazidime, chloramphenicol, and vancomycin but was intermediate to erythromycin and clindamycin. The patient recovered without sequelae after treatment with appropriate antibiotics for two weeks.
Diagnosis of Mixed
Plasmodium malariae
and
P. vivax
Infection in a Development Aid Volunteer by Examination of Bone-Marrow Specimens by Real-Time PCR
Authors: Imirzalioglu Can , Soydan Nedim , Schaller Markus , Bretzel Reinhard G. , Chakraborty Trinad , Domann Eugen
Published: June 2006

Mixed
Plasmodium malariae
and
P. vivax
infections in humans are reported very infrequently. The case of a 27-year-old male who sustained malaria quartana/tertiana caused by an unbalanced mixed
P. malariae
-
P. vivax
infection is reported here. Conventional tests and serology for malarial parasites were uniformly negative. Identification and quantification of the parasites were accomplished by examining bone-marrow specimens using specific real-time TaqMan PCR.
Antimicrobial Agent of Susceptibilities and Antiseptic Resistance Gene Distribution among Methicillin-Resistant
Staphylococcus aureus
Isolates from Patients with Impetigo and Staphylococcal Scalded Skin Syndrome
Authors: Noguchi Norihisa , Nakaminami Hidemasa , Nishijima Setsuko , Kurokawa Ichiro , So Hiromu , Sasatsu Masanori
Published: June 2006

The susceptibilities to antimicrobial agents of and distributions of antiseptic resistance genes in methicillin-resistant
Staphylococcus aureus
(MRSA) strains isolated between 1999 and 2004 in Japan were examined. The data of MRSA strains that are causative agents of impetigo and staphylococcal scalded skin syndrome (SSSS) were compared with those of MRSA strains isolated from patients with other diseases. The susceptibilities to antiseptic agents in MRSA isolates from patients with impetigo and SSSS were higher than those in MRSA isolates from patients with other diseases. The distribution of the
qacA/B
genes in MRSA strains isolated from patients with impetigo and SSSS (1.3%, 1/76) was remarkably lower than that in MRSA strains isolated from patients with other diseases (45.9%, 95/207). Epidemiologic typings of staphylococcal cassette chromosome
mec
(SCC
mec
) and pulsed-field gel electrophoresis (PFGE) showed that MRSA strains isolated from patients with impetigo and SSSS had type IV SCC
mec
(75/76), except for one strain, and 64.5% (49/76) of the strains had different PFGE types. In addition, the patterns of restriction digestion of all tested
qacA/B
plasmid in MRSA isolates having different PFGE types were identical. The results showed that a specific MRSA clone carrying
qacA/B
was not prevalent, but
qacA/B
was spread among health care-associated MRSA strains. Therefore, it was concluded that the lower distribution rate of
qacA/B
resulted in higher susceptibilities to cationic antiseptic agents in MRSA isolated from patients with impetigo and SSSS.
Plant Pathogen Forensics: Capabilities, Needs, and Recommendations
Authors: Fletcher J. , Bender C. , Budowle B. , Cobb W. T. , Gold S. E. , Ishimaru C. A. , Luster D. , Melcher U. , Murch R. , Scherm H. , Seem R. C. , Sherwood J. L. , Sobral B. W. , Tolin S. A.
Published: June 2006

A biological attack on U.S. crops, rangelands, or forests could reduce yield and quality, erode consumer confidence, affect economic health and the environment, and possibly impact human nutrition and international relations. Preparedness for a crop bioterror event requires a strong national security plan that includes steps for microbial forensics and criminal attribution. However, U.S. crop producers, consultants, and agricultural scientists have traditionally focused primarily on strategies for prevention and management of diseases introduced naturally or unintentionally rather than on responding appropriately to an intentional pathogen introduction. We assess currently available information, technologies, and resources that were developed originally to ensure plant health but also could be utilized for postintroduction plant pathogen forensics. Recommendations for prioritization of efforts and resource expenditures needed to enhance our plant pathogen forensics capabilities are presented.
Cell Wall Assembly in
Saccharomyces cerevisiae
Authors: Lesage Guillaume , Bussey Howard
Published: June 2006

An extracellular matrix composed of a layered meshwork of β-glucans, chitin, and mannoproteins encapsulates cells of the yeast
Saccharomyces cerevisiae
. This organelle determines cellular morphology and plays a critical role in maintaining cell integrity during cell growth and division, under stress conditions, upon cell fusion in mating, and in the durable ascospore cell wall. Here we assess recent progress in understanding the molecular biology and biochemistry of cell wall synthesis and its remodeling in
S. cerevisiae
. We then review the regulatory dynamics of cell wall assembly, an area where functional genomics offers new insights into the integration of cell wall growth and morphogenesis with a polarized secretory system that is under cell cycle and cell type program controls.
Defenses against Oxidative Stress inNeisseria gonorrhoeae: a System Tailored for a Challenging Environment
Authors: Seib Kate L. , Wu Hsing-Ju , Kidd Stephen P. , Apicella Michael A. , Jennings Michael P. , McEwan Alastair G.
Published: June 2006

Neisseria gonorrhoeaeis a host-adapted pathogen that colonizes primarily the human genitourinary tract. This bacterium encounters reactive oxygen and reactive nitrogen species as a consequence of localized inflammatory responses in the urethra of males and endocervix of females and also of the activity of commensal lactobacilli in the vaginal flora. This review describes recent advances in the understanding of defense systems against oxidative stress inN. gonorrhoeaeand shows that while some of its defenses have similarities to the paradigm established withEscherichia coli, there are also some key differences. These differences include the presence of a defense system against superoxide based on manganese ions and a glutathione-dependent system for defense against nitric oxide which is under the control of a novel MerR-like transcriptional regulator. An understanding of the defenses against oxidative stress inN. gonorrhoeaeand their regulation may provide new insights into the ways in which this bacterium survives challenges from polymorphonuclear leukocytes and urogenital epithelial cells.
IS
CR
Elements: Novel Gene-Capturing Systems of the 21st Century?
Authors: Toleman Mark A. , Bennett Peter M. , Walsh Timothy R.
Published: June 2006

“Common regions” (CRs), such as Orf513, are being increasingly linked to mega-antibiotic-resistant regions. While their overall nucleotide sequences show little identity to other mobile elements, amino acid alignments indicate that they possess the key motifs of IS
91
-like elements, which have been linked to the mobility
ent
plasmids in pathogenic
Escherichia coli
. Further inspection reveals that they possess an IS
91
-like origin of replication and termination sites (
ter
IS), and therefore CRs probably transpose via a rolling-circle replication mechanism. Accordingly, in this review we have renamed CRs as IS
CR
s to give a more accurate reflection of their functional properties. The genetic context surrounding IS
CR
s indicates that they can procure 5′ sequences via misreading of the cognate
ter
IS, i.e., “unchecked transposition.” Clinically, the most worrying aspect of IS
CR
s is that they are increasingly being linked with more potent examples of resistance, i.e., metallo-β-lactamases in
Pseudomonas aeruginosa
and co-trimoxazole resistance in
Stenotrophomonas maltophilia
. Furthermore, if IS
CR
elements do move via “unchecked RC transposition,” as has been speculated for IS
CR
1, then this mechanism provides antibiotic resistance genes with a highly mobile genetic vehicle that could greatly exceed the effects of previously reported mobile genetic mechanisms. It has been hypothesized that bacteria will surprise us by extending their “genetic construction kit” to procure and evince additional DNA and, therefore, antibiotic resistance genes. It appears that IS
CR
elements have now firmly established themselves within that regimen.
Regulation of the Cell Cycle by Protein Phosphatase 2A in
Saccharomyces cerevisiae
Authors: Jiang Yu
Published: June 2006

Protein phosphatase 2A (PP2A) has long been implicated in cell cycle regulation in many different organisms. In the yeast
Saccharomyces cerevisiae
, PP2A controls cell cycle progression mainly through modulation of cyclin-dependent kinase (CDK) at the G
2
/M transition. However, CDK does not appear to be a direct target of PP2A. PP2A affects CDK activity through its roles in checkpoint controls. Inactivation of PP2A downregulates CDK by activating the morphogenesis checkpoint and, consequently, delays mitotic entry. Defects in PP2A also compromise the spindle checkpoint and predispose the cell to an error-prone mitotic exit. In addition, PP2A is involved in controlling the G
1
/S transition and cytokinesis. These findings suggest that PP2A functions in many stages of the cell cycle and its effect on cell cycle progression is pleiotropic.
Studies on Improved Synthesis of 2'-Deoxyribonucleosides of Pyridazine Derivatives
Authors: Kazimierczuk Zygmunt , Kamiński Jarosław , Meggio Flavio
Published: June 2006

A number of 2'-deoxyribonucleosides of halogenated pyridazine derivatives were prepared by glycosylation of their respective potassium or DBU salts in acetone. The reaction yielded predominatly β-anomers that could be purified by simple crystallization or column chromatography. Of the studied pyridazines and deoxynucleosides, only 4-bromo-6-chloropyridazin-3-one and 6-chloro-2-(2'-deoxyribofuranosyl)pyridazin-3-one showed modest inhibition of CK2 kinase.
Effect of a Hydrogen Bonding Carboxamide Group on Universal Bases
Authors: Too Kathleen , Brown Daniel M. , Holliger Philipp , Loakes David
Published: June 2006

A number of aromatic universal base analogues have been described in the literature, but most are non-hydrogen bonding. We have examined the effect of introducing hydrogen bonding carboxamide groups onto the pyrrole ring of 5-nitroindole. The modified analogues retain universal base features, but there are no overall effects on duplex stability. This leads to the suggestion that the nitro group is within the hydrogen bonding face of the duplex, and the hydrogen bonding carboxamide group is in the duplex major groove.
Exposure to non-arsenic pesticides is associated with lymphoma among farmers in Spain
Authors: van Balen E , Font R , Cavallé N , Font L , Garcia-Villanueva M , Benavente Y , Brennan P , de Sanjose S
Published: June 2006

Objectives:To estimate the risk of lymphoma among farmers in Spain.

Methods:This is a multicentre case control study conducted in Spain. Cases were subjects diagnosed with lymphoma according to the World Health Organization (WHO) classification in four hospitals between 1998–2002. Hospital controls were frequency matched to the cases by sex, age, and centre. All subjects were interviewed about jobs ever held in lifetime for at least one year and the exposures in those jobs were recorded. The risk of lymphomas among subjects ever having had a job as a farmer was compared with all other occupations. Farmers were analysed according to the type of farming job performed: crop farming, animal farming, and general farming. Occupational exposure was summarised into 15 main categories: organic dust, radiation, contact with animals, PAH, non-arsenic pesticides (carbamates, organophosphates, chlorinated hydrocarbons, triazines and triazoles, phenoxy herbicides, chlorophenols, dibenzodioxin, and dibenzofuran), arsenic pesticides, contact with meat, contact with children, solvents, asbestos, soldering fumes, organic colourants, polychlorinated biphenyls, ethylene oxide, and hair dyes.

Results:Although farmers were not at an increased risk of lymphoma as compared with all other occupations, farmers exposed to non-arsenic pesticides were found to be at increased risk of lymphoma (OR = 1.8, 95% CI 1.1 to 2). This increased risk was observed among farmers working exclusively either as crop farmers or as animal farmers (OR = 2.8, 95% CI 1.3 to 5.8). Risk was highest for exposure to non-arsenic pesticides for over nine years (OR = 2.4, 95% CI 1.2 to 2.8).

Conclusions:Long term exposure to non-arsenic pesticides may induce lymphomagenesis among farmers.
Wheezing, asthma, hay fever, and atopic eczema in relation to maternal occupations in pregnancy
Authors: Magnusson L L , Wennborg H , Bonde J P , Olsen J
Published: June 2006

Objectives:To examine whether prenatal occupational exposures, especially to organic solvents, are associated with atopic diseases in childhood.

Methods:The study comprised children born in Odense or Aalborg, Denmark between 1984 and 1987. Occupational job titles were derived from questionnaires filled out by the mothers when attending midwife centres. Assessment of organic solvent exposures was based on job titles selected by occupational specialists. A follow up questionnaire to the parents provided data on medical diagnoses as well as wheezing symptoms for 7844 children aged 14–18. Multivariate logistic regression analyses were performed to estimate the cumulative risk for wheezing (early wheezing not diagnosed as asthma), asthma, hay fever, and atopic eczema during childhood by means of odds ratios (OR) and 95% confidence intervals (CI).

Results:Explorative analyses by maternal job titles in pregnancy showed elevated odds ratios concerning different atopic diseases for occupational groups such as “bakers, pastry cooks, and confectionary makers”, “dental assistants”, “electrical and electronic assemblers”, “sewers and embroiders”, and “bookbinders and related workers”. An excess risk ratio for hay fever (OR 2.8, CI 1.1 to 7.5) was found following maternal gestational exposure to organic solvents. Furthermore, a slightly raised odds ratio for asthma was observed in children of shift workers (OR 1.2, CI 1.0 to 1.5).

Conclusion:The data suggest links between certain maternal occupations during pregnancy and atopic diseases, which merits further scrutiny. However, no consistent pattern was seen across the different atopic diseases.