bluuebunny/crossref_metadata_2025 huggingface dataset:
Contact Activation in Low‐density Lipoprotein Apheresis Systems
Authors: Krieter Detlef H. , Steinke Jörn , Kerkhoff Markus , Fink Edwin , Lemke Horst‐Dieter , Zingler Christine , Müller Gerhard A. , Schuff‐Werner Peter
Published: January 2005

Abstract: Background: Anaphylactoid reactions due to contact activation have been observed in patients on ACE inhibitor therapy and hemodialysis with negatively charged dialysis membranes. Negatively charged surfaces are functional constituents of different LDL apheresis systems. Therefore, contact activation was investigated during LDL apheresis with three different systems: (i) heparin‐induced extracorporeal LDL precipitation (HELP); (ii) dextran sulfate cellulose (DSC) columns; and (iii) modified polyacrylate gels (DALI) in a clinical setting. Methods: 24 prevalent patients on regular LDL apheresis treatment were included in the study. Bradykinin, prekallikrein, and HMW kininogen were measured during a single LDL apheresis at different sites of the systems. Results: LDL apheresis with DSC and DALI was associated with an extreme release of bradykinin after the passage of plasma or blood through the LDL adsorbers as well as with a decrease of prekallikrein and HMW kininogen during the course of the treatment. Bradykinin release exceeded the degradation capacity of the kininase II, since markedly elevated bradykinin concentrations were observed in the arterial line of the extracorporeal circuits of both systems. This was not associated with anaphylactoid reactions. In contrast to the treatments with DSC and DALI, the HELP system did not lead to any activation of the kallikrein–kinin system. Conclusion: From our data we conclude that angiotensin converting enzyme (ACE) inhibitors are contraindicated in patients on LDL apheresis with the DSC and the DALI system. Because the HELP system does not activate the kallikrein–kinin system, patients who need ACE inhibitors are predisposed for this LDL apheresis procedure.
Pancreatic Polypeptide Administration Reduces Insulin Requirements of Artificial Pancreas in Pancreatectomized Dogs
Authors: Kono Tetsuya , Hanazaki Kazuhiro , Yazawa Kazuyuki , Ashizawa Satoshi , Fisher William E. , Wang Xiao‐Ping , Nosé Yukihiko , Brunicardi F. Charles
Published: January 2005

Abstract:  An artificial endocrine pancreas is a mechanical device that frequently measures blood glucose and adjusts the rate of insulin infusion to maintain normoglycemia. In this study, we evaluated the effect of pancreatic polypeptide (PP) on insulin requirements after total pancreatectomy. However, other endocrine hormones are needed not only to facilitate the effect of insulin, but also to regulate insulin functions in vivo. In this study, the effect of PP infusion on insulin requirements after total pancreatectomy in dogs is examined. After total pancreatectomy, five dogs were supported by artificial endocrine pancreas model STG‐22 for 72 h. In a second group of five dogs, both insulin and PP were infused. Mean blood glucose levels and insulin requirements were compared between the two groups. There was no difference in mean plasma glucose levels between the two groups. In all 10 dogs, the mean blood glucose level for 72 h was 110 ± 4 mg/dL and was tightly controlled between 65 and 190 mg/dL. However, the insulin requirement for the first and second postoperative days in the group treated with PP was significantly less than that of the control group (90.0 ± 20.8 mU/kg vs. 445.0 ± 151.9 mU/kg; P < 0.05, and 562.7 ± 126.5 mU/kg vs. 1007.7 ± 144.9 mU/kg; P < 0.05, respectively). We conclude that infusion of PP reduces the insulin requirement for the initial 48 h in pancreatectomized dogs treated with an artificial endocrine pancreas.
Erythrocyte Susceptibility to Oxidative Stress in Chronic Renal Failure Patients Under Different Substitutive Treatments
Authors: Lucchi Leonardo , Bergamini Stefania , Iannone Anna , Perrone Salvatore , Stipo Lucia , Olmeda Fabio , Caruso Francesco , Tomasi Aldo , Albertazzi Alberto
Published: January 2005

Abstract:  An increased oxidative stress is now considered one of the major risk factors in chronic renal failure (CRF) patients that may be exacerbated by dialysis. It has been postulated that this increased oxidative stress might cause an augmented red blood cell (RBC) membrane lipid peroxidation with the consequent alteration in membrane deformability. The aim of this study was to evaluate RBC susceptibility to an in vitro induced oxidative stress and RBC antioxidant potential in different groups of CRF patients undergoing different substitutive treatment modalities. Fifteen end‐stage CRF patients were evaluated in conservative treatment, 23 hemodialysis (HD) patients, 15 continuous ambulatory peritoneal dialysis (CAPD) patients, 15 kidney transplanted patients, and 16 controls. Their RBCs were incubated with the oxidative stress‐inducing agent tert‐butylhydroperoxide both in the presence and in the absence of the catalase inhibitor sodium azide, and the level of malondialdehyde (MDA) (a product of lipid peroxidation), was measured at 0, 5, 10, 15, and 30 min of incubation. In addition, the RBC content of reduced glutathione (GSH) was measured by HPLC. As opposed to the controls, RBCs from end‐stage CRF patients exhibited an increased sensitivity to oxidative stress induced in vitro, both in the absence and presence of a catalase inhibitor, as demonstrated by a significantly higher level of MDA production at all the incubation times (P < 0.05). Different substitutive treatments had different impacts on this phenomenon; CAPD and kidney transplantation were able to normalize this alteration while HD was not. GSH appeared to be related to the increase in RBC susceptibility to oxidative stress; its content being significantly elevated in end‐stage CRF and HD patients as compared with CAPD and transplanted patients and controls (P < 0.05). No significant changes were observed in the RBC glutathione content during the HD session. The increase of GSH in RBCs of end‐stage CRF and HD patients seems to indicate the existence of an adaptive mechanism under increased oxidative stress occurring in vivo. Unlike HD, the beneficial effect of CAPD on the anemia of dialysis patients might partly be due to a condition of lower oxidative stress that might in addition counterbalance the cardiovascular negative effects of dislipidemia  of  CAPD  patients.
Intra‐aortic Balloon Pumping: Does Posture Matter?
Authors: Khir Ashraf W. , Price Susana , Hale Colin , Young David A. , Parker Kim H. , Pepper John R.
Published: January 2005

Abstract:  Background: The effect of the posture of semirecumbent patients on the hemodynamics and performance of intra‐aortic balloon (IAB) were studied in vivo and in vitro. Methods: An IAB was inserted into a glass tube filled with saline, fully inflated and deflated using an intra‐aortic balloon pump (IABP). Three successive cycles were filmed at 125 frames/s with the tube positioned at various angles between 0° (horizontal) and 90° (vertical). Pressure and flow were measured distal to both ends of the balloon. In parallel, coronary left anterior descending (LAD) flow velocity and aortic pressure were recorded in 6 patients using IABP, postcardiac surgery in the intensive care unit. Recordings were made when the patient was lying horizontally (recumbent) and when the patient's torso was inclined at 30° to the horizontal (semirecumbent). Results: With the tube horizontal, the inflation was effectively uniform along the length of the balloon. At all other angles, the higher end of the balloon inflated first, and mean pressure and flow measured distal to the higher end of the balloon were less than those measured at 0°. Mean aortic pressure and LAD flow decreased by 10 ± 2% (P = 0.001) and 15 ± 3% (P = 0.001), respectively, when the patient was semirecumbent compared to when the patient was recumbent. Conclusion: The decreased mean aortic pressure and LAD flow velocity suggests that unless patients using IABP are required to be semirecumbent, it may be best to position them horizontally to gain the full benefits of balloon counter pulsation to the coronary circulation.
Drop outs from in‐patient treatment of anorexia nervosa
Authors: Zeeck A. , Hartmann A. , Buchholz C. , Herzog T.
Published: January 2005

Objective:  Aim of the study was to identify patient characteristics that distinguish drop outs and completers from in‐patient treatment for anorexia nervosa.

Method:  A total of 133 consecutively admitted in‐patients with anorexia nervosa (age range 16–50 years; 92.5% women) were analysed using sociodemographic variables as well as measures of psychopathology (SCL‐90‐R, EDI‐2) and interpersonal difficulties (IIP‐C). Patients were treated in a multimodal treatment setting, combining cognitive‐behavioural and psychodynamic components.

Results:  Patients, who reported fewer symptoms, were hospitalized before and had a comorbid depression stayed more often in psychotherapy. Patients dropping out of treatment (31.6%) showed a trend to higher levels of maturity fears. Subtype, age, duration of illness, comorbid personality disorders or previous drop outs were not predictive of dropping out.

Conclusion:  Addressing the high ambivalence and maturity fears of anorexic patients should be an essential issue in psychotherapy with this patient group.
General psychopathology is more important for executive functioning than diagnosis
Authors: Stordal K. I. , Mykletun A. , Asbjørnsen A. , Egeland J. , Landrø N. I. , Roness A. , Rund B. R. , Sundet K. S. , Lundervold A. J. , Lund A.
Published: January 2005

Objective:  Impaired executive functioning (EF) has often been reported in patients with major depression or schizophrenia. We hypothesize that the variance in EF is more affected by level of general psychopathology than by diagnosis.

Method:  Forty‐three patients with major depression and 47 with schizophrenia were included. EF was measured with Wisconsin Card Sorting Test, Stroop Colour Word Test, Paced Auditory Serial Addition Test, Digits Backwards and Controlled Oral Word Association Test. The level of general psychopathology was measured with Brief Psychiatric Rating Scale – Expanded and Positive and Negative Syndrome Scale, the General psychopathology subscale.

Results:  The level of general psychopathology predicted more of the variance in EF than diagnosis. In multivariate analyses, the effect of general psychopathology on EF was more robust for adjustment for diagnosis than vice versa.

Conclusion:  Future research on cognitive functioning in psychiatric patients should include level of general psychopathology to avoid overemphasising effects of diagnoses.
A right orbitofrontal region and OCD symptoms: a case report
Authors: Ogai M. , Iyo M. , Mori N. , Takei N.
Published: January 2005

Objective:  To discuss the relationship between obsessive‐compulsive symptoms and a right orbitofrontal lesion.

Method:  Single case report.

Results:  A 59‐year‐old man developed obsessive‐compulsive disorder (OCD) symptoms after his head injury. Magnetic resonance imaging brain scans showed a small contusion in the right orbitofrontal region, and single‐photon emission computed tomography revealed hypoperfusion in blood flow at the same region.

Conclusion:  The OCD symptoms that developed in the present case may be attributable primarily to hypofunction in the lesion localized to the right orbitofrontal area. Although caution is needed for interpretation of the observation because of our experience of only a single case, it suggests that the right orbitofrontal region may be important in forming OCD symptoms.
Help‐seeking behaviour in patients with anxiety disorder and depression
Authors: Roness A. , Mykletun A. , Dahl A. A.
Published: January 2005

Objective:  The objective of this study was to investigate help‐seeking behaviour among persons with anxiety disorder and depression based on self‐rating in a Norwegian population (the HUNT study).

Method:  Of the 92 100 inhabitants aged 20–89 years invited, 65 648 (71.3%) took part. Among them 60 869 (66.1%) persons delivered valid ratings on hospital anxiety and depression scale, and had answered the requested help for mental problems question.

Results:  Among HUNT attenders 13% of those with depression and 25% with anxiety disorders had been help seekers. Help seeking was only non‐significantly associated with demographic or other variables.

Conclusion:  Most persons with anxiety disorder and/or depression in the population had not sought help for their mental disorders, but the disparity between use and need of health service must not be overassessed. Improvement of the help‐seeking rate for common mental disorder should have high priority in mental health politics.
Have there been any changes in the public's attitudes towards psychiatric treatment? Results from representative population surveys in Germany in the years 1990 and 2001
Authors: Angermeyer M. C. , Matschinger H.
Published: January 2005

Objective:  During the 1990s, the reform of German mental health care made considerable progress. In addition, important advances in the treatment of mental disorders took place. The question arises as to whether these improvements were accompanied by a greater acceptance of psychiatric treatment by the public.

Method:  In 1990 and 2001, representative surveys were conducted among the adult German population, using the same sampling procedure and interview.

Results:  In the course of the 1990s, the German public became more inclined to recommend to seek help from psychiatrists or psychotherapists in case of schizophrenia or major depression. There was also an increase in the willingness to recommend therapy in general, which was particularly pronounced with regard to drug treatment and psychotherapy of schizophrenia.

Conclusion:  Our findings seem to support the notion that improvements in mental health care are reflected in the public's attitudes towards psychiatric treatment. This may, in turn, positively impact help‐seeking behaviour.
Multigene Phylogenetic Analysis of Pathogenic
Candida
Species in the
Kazachstania
(
Arxiozyma
)
telluris
Complex and Description of Their Ascosporic States as
Kazachstania bovina
sp. nov.,
K. heterogenica
sp. nov.,
K. pintolopesii
sp. nov., and
K. slooffiae
sp. nov
Authors: Kurtzman Cletus P. , Robnett Christie J. , Ward Jerrold M. , Brayton Cory , Gorelick Peter , Walsh Thomas J.
Published: January 2005

A yeast causing widespread infection of laboratory mice was identified from 26S rRNA gene sequences as
Candida pintolopesii
. To determine the relationship of
C. pintolopesii
with other members of the
Kazachstania
(
Arxiozyma
)
telluris
species complex, nucleotide sequences from domains 1 and 2 of the 26S rRNA gene, the mitochondrial small-subunit rRNA gene, and the RNA polymerase II gene were phylogenetically analyzed. That analysis resolved the 48 strains examined into five closely related species:
K. telluris
,
Candida bovina
,
C. pintolopesii
,
Candida slooffiae
, and a previously unknown species. One or more strains of each of the last four species formed an ascosporic state much like that of
K. telluris
. To place these ascosporogenous strains taxonomically, it is proposed that they be assigned to the teleomorphic genus
Kazachstania
as
K. bovina
(type strain NRRL Y-7283, CBS 9732, from the nasal passage of a pigeon),
K. heterogenica
(type strain NRRL Y-27499, CBS 2675, from rodent feces),
K. pintolopesii
(type strain NRRL Y-27500, CBS 2985, from the peritoneal fluid of a dead guinea pig), and
K. slooffiae
(type strain NRRL YB-4349, CBS 9733, from the cecum of a horse). On the basis of multigene sequence analyses,
K. heterogenica
appears to be a hybrid of
K. pintolopesii
and a presently unknown species. With the exception of
K. bovina
, the phylogenetically defined species show a moderate degree of host specificity.
Preclinical Diagnosis of African Swine Fever in Contact-Exposed Swine by a Real-Time PCR Assay
Authors: Zsak L. , Borca M. V. , Risatti G. R. , Zsak A. , French R. A. , Lu Z. , Kutish G. F. , Neilan J. G. , Callahan J. D. , Nelson W. M. , Rock D. L.
Published: January 2005

A fluorogenic probe hydrolysis (TaqMan) PCR assay for African swine fever virus (ASFV) was developed and evaluated in experimentally infected swine. This sensitive and specific one-step single-tube assay, which can be performed in 2 h or less, detected viral DNA in tonsil scraping samples 2 to 4 days prior to onset of clinical disease. Thus, the assay would have application for preclinical diagnosis of African swine fever and surveillance and/or emergency management of a disease outbreak.
Association between Respiratory Disease and Bacterial and Viral Infections in British Racehorses
Authors: Wood J. L. N. , Newton J. R. , Chanter N. , Mumford J. A.
Published: January 2005

Respiratory disease is important in horses, particularly in young Thoroughbred racehorses, and inflammation that is detected in the trachea and bronchi (termed inflammatory airway disease [IAD]) is more significant in this population in terms of impact and frequency than other presentations of respiratory disease. IAD, which is characterized by neutrophilic inflammation, mild clinical signs, and accumulation of mucus in the trachea, may be multifactorial, possibly involving infections and environmental and immunological factors, and its etiology remains unclear. This 3-year longitudinal study of young Thoroughbred racehorses was undertaken to characterize the associations of IAD and nasal discharge with viral and bacterial infections. IAD was statistically associated with tracheal infection withStreptococcus pneumoniae(capsule type 3),Streptococcus zooepidemicus,Actinobacillusspp., andMycoplasma equirhinisand equine herpesvirus 1 and 4 infections, after adjustment for variation between training yards, seasons, and age groups. The association withS. pneumoniaeandS. zooepidemicuswas independent of prior viral infection and, critically, was dependent on the numbers of organisms isolated.S. pneumoniaewas significant only in horses that were 2 years old or younger. The prevalence and incidence of IAD,S. zooepidemicus, andS. pneumoniaedecreased in parallel with age, consistent with increased disease resistance, perhaps by the acquisition of immunity. The study provided evidence forS. zooepidemicusandS. pneumoniaeplaying an important etiological role in the pathogenesis of IAD in young horses.
Risk Factors and Molecular Analysis of Community Methicillin-Resistant
Staphylococcus aureus
Carriage
Authors: Lu Po-Liang , Chin Lien-Chun , Peng Chien-Fang , Chiang Yi-Hsiung , Chen Tyen-Po , Ma Ling , Siu L. K
Published: January 2005

A total of 1,838 subjects from the community and 393 subjects from health care-related facilities in Taiwan were evaluated for the prevalence of nasal
Staphylococcus aureus
colonization and to identify risk factors associated with
S. aureus
and methicillin-resistant
S. aureus
(MRSA) colonization. Among the community subjects, 3.5% had nasal MRSA colonization. Subjects from health care-related facilities had a lower
S. aureus
colonization rate (19.1%) than community subjects (25.2%) but had a significantly higher rate of colonization with MRSA (7.63%). Age (
P
< 0.001) was a significant risk factor for
S. aureus
colonization, with subjects under age 20 years or between 71 and 80 years showing higher rates of colonization. Recent gastrointestinal disease (
P
= 0.011) and hospital admission (
P
= 0.026) were risk factors for nasal MRSA colonization. Comparison of hospital MRSA isolates with the colonization strains by staphylococcal cassette chromosome
mec
(SCC
mec
) gene typing and pulsed-field gel electrophoresis (PFGE) typing revealed that most MRSA strains carried in the community were SCC
mec
type IV and that most clinical hospital isolates were type III, while health care facility-related carriage isolates were mainly SCC
mec
type III and type IV. Two new variant SCC
mec
types were identified. Six clusters of PFGE patterns were distinguished: two mainly comprised health care facility-related MRSA strains, three mainly comprised community MRSA strains, and one comprised mixed community and health care facility-related MRSA strains. In conclusion, a high prevalence of MRSA colonization was observed among people with no relationship to the hospital setting. The high level of multiple-drug resistance among community MRSA strains in association with the previously reported excessive use of antibiotics in Taiwan highlights the importance of the problem of antibiotic selective pressure. Our results indicate that both the clonal spread of MRSA and the transmission of hospital isolates contribute to the high MRSA burden in the community.
Interlaboratory Comparison of PCR-Based Identification and Genogrouping of
Neisseria meningitidis
Authors: Taha Muhamed-Kheir , Alonso Jean-Michel , Cafferkey Mary , Caugant Dominique A. , Clarke Stuart C. , Diggle Mathew A. , Fox Andrew , Frosch Matthias , Gray Stephen J. , Guiver Malcolm , Heuberger Sigrid , Kalmusova Jitka , Kesanopoulos Konstantinos , Klem Anne-Marie , Kriz Paula , Marsh John , Mölling Paula , Murphy Karen , Olcén Per , Sanou Oumar , Tzanakaki Georgina , Vogel Ulrich
Published: January 2005

Twenty clinical samples (18 cerebrospinal fluid samples and 2 articular fluid samples) were sent to 11 meningococcus reference centers located in 11 different countries. Ten of these laboratories are participating in the EU-MenNet program (a European Union-funded program) and are members of the European Monitoring Group on Meningococci. The remaining laboratory was located in Burkina Faso.
Neisseria meningitidis
was sought by detecting several meningococcus-specific genes (
crgA
,
ctrA
, 16S rRNA, and
porA
). The PCR-based nonculture method for the detection of
N. meningitidis
gave similar results between participants with a mean sensitivity and specificity of 89.7 and 92.7%, respectively. Most of the laboratories also performed genogrouping assays (
siaD
and
mynB
/
sacC
). The performance of genogrouping was more variable between laboratories, with a mean sensitivity of 72.7%. Genogroup B gave the best correlation between participants, as all laboratories routinely perform this PCR. The results for genogroups A and W135 were less similar between the eight participating laboratories that performed these PCRs.
Characterization of
ermB
Gene Transposition by Tn
1545
and Tn
917
in Macrolide-Resistant
Streptococcus pneumoniae
Isolates
Authors: Okitsu Naohiro , Kaieda Satoru , Yano Hisakazu , Nakano Ryuichi , Hosaka Yoshio , Okamoto Ryouichi , Kobayashi Toshimitsu , Inoue Matsuhisa
Published: January 2005

In
Streptococcus pneumoniae
, the
ermB
gene is carried by transposons, such as Tn
917
and Tn
1545
. This study investigated the relationship between macrolide resistance and the presence of the
ermB
gene on Tn
917
or Tn
1545
in 84 Japanese pneumococcal isolates. Macrolide-resistant strains were classified into two groups as follows. Group 1 (19 strains) showed a tendency to high resistance to erythromycin (MIC at which 50% of isolates are inhibited, 4 mg/liter; MIC at which 90% of isolates are inhibited [MIC
90
], 128 mg/liter) but susceptibility to rokitamycin (MIC
90
, 1 mg/liter), with the
ermB
gene located on Tn
1545
. Group 2 (65 strains) showed a tendency to high resistance to both antibiotics (MIC
90
s for both erythromycin and rokitamycin, >128 mg/liter), with the
ermB
gene located on Tn
917
. There were no strains with constitutive macrolide resistance in either group. All of the strains in group 2 had a deletion in the promoter region of
ermB
and an insertion of the TAAA motif in the leader peptide. The results of pulsed-field gel electrophoresis and serogrouping showed that Tn
1545
spread clonally while Tn
917
spread both horizontally and clonally. In conclusion, in Japanese macrolide-resistant
S. pneumoniae
isolates, the
ermB
gene is carried and spread primarily by Tn
917
.
Benefit of Hepatitis C Virus Core Antigen Assay in Prediction of Therapeutic Response to Interferon and Ribavirin Combination Therapy
Authors: Takahashi Masahiko , Saito Hidetsugu , Higashimoto Makiko , Atsukawa Kazuhiro , Ishii Hiromasa
Published: January 2005

A highly sensitive second-generation hepatitis C virus (HCV) core antigen assay has recently been developed. We compared viral disappearance and first-phase kinetics between commercially available core antigen (Ag) assays, Lumipulse Ortho HCV Ag (Lumipulse-Ag), and a quantitative HCV RNA PCR assay, Cobas Amplicor HCV Monitor test, version 2 (Amplicor M), to estimate the predictive benefit of a sustained viral response (SVR) and non-SVR in 44 genotype 1b patients treated with interferon (IFN) and ribavirin. HCV core Ag negativity could predict SVR on day 1 (sensitivity = 100%, specificity = 85.0%, accuracy = 86.4%), whereas RNA negativity could predict SVR on day 7 (sensitivity = 100%, specificity = 87.2%, accuracy = 88.6%). None of the patients who had detectable serum core Ag or RNA on day 14 achieved SVR (specificity = 100%). The predictive accuracy on day 14 was higher by RNA negativity (93.2%) than that by core Ag negativity (75.0%). The combined predictive criterion of both viral load decline during the first 24 h and basal viral load was also predictive for SVR; the sensitivities of Lumipulse-Ag and Amplicor-M were 45.5 and 47.6%, respectively, and the specificity was 100%. Amplicor-M had better predictive accuracy than Lumipulse-Ag in 2-week disappearance tests because it had better sensitivity. On the other hand, estimates of kinetic parameters were similar regardless of the detection method. Although the correlations between Lumipulse-Ag and Amplicor-M were good both before and 24 h after IFN administration, HCV core Ag seemed to be relatively lower 24 h after IFN administration than before administration. Lumipulse-Ag seems to be useful for detecting the HCV concentration during IFN therapy; however, we still need to understand the characteristics of the assay.
Characterization of Sucrose-Negative
Pasteurella multocida
Variants, Including Isolates from Large-Cat Bite Wounds
Authors: Christensen Henrik , Bisgaard Magne , Angen Øystein , Frederiksen Wilhelm , Olsen John Elmerdahl
Published: January 2005

To validate the identification of
Pasteurella multocida-
like bacteria negative for acid formation from sucrose, including isolates from bite wounds caused by large cats, 17 strains were phenotypically and genotypically characterized. Phylogenetic analysis of partially sequenced
rpoB
and
infB
genes showed the monophyly of the strains characterized and the reference strains of
P. multocida
. The sucrose-negative strains formed two groups, one related to reference strains of
P. multocida
and the other related to a separate species-like group (taxon 45 of Bisgaard). DNA-DNA hybridization further documented the species-like nature of this group. Ribotyping showed the heterogeneity of all strains except four strains that shared the same ribotype and that were isolated from bovine lungs. Phylogenetic analysis by 16S rRNA sequence comparison showed the monophyly of the strains characterized and the reference strains of
P. multocida
. Two strains isolated from leopard bite wounds were related to the type strain of
P. dagmatis
; however, they represented a new taxon (taxon 46 of Bisgaard), in accordance with their distinct phenotypic and genotypic identifications. The present study documents that sucrose-negative strains of
P. multocida-
like bacteria belong to two genotypically distinct groups. The study further confirms the phenotypic heterogeneity of
P. multocida
strains and documents two new species-like taxa of
Pasteurella
related to
P. multocida
. Until diagnostic tools have been further elaborated, special care should be taken in the identification of
Pasteurella
-like bacteria isolated from bite wounds caused by large cats. The evidence of phenotypic and genotypic divergence calls for the further development of PCR tests and DNA sequencing to document doubtful isolates.
Candida orthopsilosis
and
Candida metapsilosis
spp. nov. To Replace
Candida parapsilosis
Groups II and III
Authors: Tavanti Arianna , Davidson Amanda D. , Gow Neil A. R. , Maiden Martin C. J. , Odds Frank C.
Published: January 2005

Two new species,
Candida orthopsilosis
and
C. metapsilosis
, are proposed to replace the existing designations of
C. parapsilosis
groups II and III, respectively. The species
C. parapsilosis
is retained for group I isolates. Attempts to construct a multilocus sequence typing scheme to differentiate individual strains of
C. parapsilosis
instead revealed fixed DNA sequence differences between pairs of subgroups in four genes:
COX3
,
L1A1
,
SADH
, and
SYA1
. PCR amplicons for sequencing were obtained for these four plus a further seven genes from 21 group I isolates. For nine group II isolates, PCR products were obtained from only 5 of the 11 genes, and for two group III isolates PCR products were obtained from a different set of 5 genes. Three of the PCR products from group II and III isolates differed in size from the group I products. Cluster analysis of sequence polymorphisms from
COX3
,
SADH
, and
SYA1
, which were common to the three groups, consistently separated the isolates into three distinct sets. All of these differences, together with DNA sequence similarities <90% in the
ITS1
sequence, suggest the subgroups should be afforded species status. The near absence of DNA sequence variability among isolates of
C. parapsilosis
and relatively high levels of sequence variability among isolates of
C. orthopsilosis
suggest that the former species may have evolved very recently from the latter.
Sequence Typing and Comparison of Population Biology of
Campylobacter coli
and
Campylobacter jejuni
Authors: Dingle Kate E. , Colles Frances M. , Falush Daniel , Maiden Martin C. J.
Published: January 2005

A multilocus sequence typing (MLST) scheme that uses the same loci as a previously described system for
Campylobacter jejuni
was developed for
Campylobacter coli
. The
C. coli-
specific primers were validated with 53 isolates from humans, chickens, and pigs, together with 15 Penner serotype reference isolates. The nucleotide sequence of the
flaA
short variable region (SVR) was determined for each isolate. These sequence data were compared to equivalent information for 17
C. jejuni
isolates representing the known genetic diversity of this species.
C. coli
and
C. jejuni
share approximately 86.5% identity at the nucleotide sequence level within the MLST loci. There is evidence of genetic exchange of the housekeeping genes between the two species, but at a very low rate; only one sequence type from each species showed evidence of imported DNA. The
flaA
gene was more variable and has been exchanged many times between the two species, making it an unreliable marker for species identification but useful for distinguishing closely related strains. All but 3 of 21 human
C. coli
clinical isolates were distinct, according to the combined MLST and SVR sequences. The use of a common MLST scheme allows direct comparisons of the population biology and molecular epidemiology of these two closely related human pathogens.
Quantitative Multiplex Assay for Simultaneous Detection and Identification of Indiana and New Jersey Serotypes of Vesicular Stomatitis Virus
Authors: Rasmussen Thomas B. , Uttenthal Åse , Fernández Jovita , Storgaard Torben
Published: January 2005

In order to establish a rapid and reliable system for the detection of vesicular stomatitis virus (VSV), we developed a quantitative reverse transcription-PCR assay for the detection, quantification, and differentiation of the major serotypes, VSV Indiana and VSV New Jersey, using a closed-tube multiplex format. The detection system is based on the recently invented primer-probe energy transfer (PriProET) system. A region of the gene encoding the RNA-dependent RNA polymerase was amplified by using VSV-specific primers in the presence of two serotype-specific fluorescent probes. By incorporating nucleotide analogues in the primers, both serotypes were amplified with similar efficiencies. The generation of specific amplicons resulted in fluorescent signals for either of the two serotypes, and the specificities of the reactions were confirmed from the melting temperature profiles of the fluorescent probes. The limits of detection were found to be less than 10 50% tissue culture infective doses/ml for both serotypes. The diagnostic value of the new method was tested with clinical materials from experimentally infected pigs, and it is concluded that the method is a powerful tool for the rapid identification of VSV.
Methicillin-Resistant
Staphylococcus caprae
in a Neonatal Intensive Care Unit
Authors: Ross T. L. , Fuss E. P. , Harrington S. M. , Cai M. , Perl T. M. , Merz W. G.
Published: January 2005

Staphylococcus caprae
, a hemolytic coagulase-negative staphylococcus that is infrequently associated with humans, was initially detected in specimens from six infants in our neonatal intensive care unit due to phenotypic characteristics common to methicillin-resistant
Staphylococcus aureus
. These isolates were subsequently identified as
S. caprae
by the Automated RiboPrinter microbial characterization system. This prompted an 8-month retrospective investigation in our neonatal intensive care unit.
S. caprae
was the cause of 6 of 18 episodes of coagulase-negative staphylococcal bacteremia, was the most common coagulase-negative staphylococcus recovered from the nares of 6 of 32 infants surveyed in a methicillin-resistant
S. aureus
surveillance program, and was isolated from 1 of 37 health care providers' hands. Of 13 neonatal intensive care unit isolates tested, all were methicillin resistant and positive for the
mecA
gene. All 21 isolates were found to be a single strain by Automated RiboPrinter and pulsed-field gel electrophoresis with ApaI or SmaI digestion; ApaI was more discriminating in analyzing epidemiologically unrelated strains than Automated RiboPrinter or electrophoresis with SmaI. These findings extend the importance of
S. caprae
, emphasize its similarities to methicillin-resistant
S. aureus
, and demonstrate its ability to persist in an intensive care unit setting.
Genetic Typing of the Porin Protein of
Neisseria gonorrhoeae
from Clinical Noncultured Samples for Strain Characterization and Identification of Mixed Gonococcal Infections
Authors: Lynn Freyja , Hobbs Marcia M. , Zenilman Jonathan M. , Behets Frieda M. T. F. , Van Damme Kathleen , Rasamindrakotroka Andry , Bash Margaret C.
Published: January 2005

Molecular methods that characterize the
Neisseria gonorrhoeae
porin protein Por are needed to study gonococcal pathogenesis in the natural host and to classify strains from direct clinical samples used with nucleic acid amplification-based tests. We have defined the capabilities of
por
variable region (VR) typing and determined suitable conditions to apply the method to direct clinical specimens. Nested PCR from spiked urine samples detected 1 to 10 copies of template DNA; freezing spiked whole urine greatly reduced the ability to amplify
porB
. In a laboratory model of mixed gonococcal infections, the
por
type of one strain could be determined in the presence of a 100-fold excess of another.
por
VR typing was used to examine clinical samples from women enrolled in studies conducted in Baltimore, Md., and Madagascar.
por
type was determined from 100% of paired cervical swab and wick samples from 20 culture-positive women from Baltimore; results for eight individuals (40%) suggested infection with more than one strain. In frozen urine samples from Madagascar,
porB
was amplified and typed from 60 of 126 samples from ligase chain reaction (LCR)-positive women and 3 samples from LCR-negative women. The
por
VR types of 13 samples (21%) suggested the presence of more than one gonococcal strain. Five
por
types, identified in >45% of women with typed samples, were common to both geographic areas. Molecular typing is an important adjunct to nucleic acid amplification-based diagnostics. Methods that utilize direct clinical samples and can identify mixed infections may contribute significantly to studies of host immunity, gonococcal epidemiology, and pathogenesis.
Latency-Related Gene Encoded by Bovine Herpesvirus 1 Promotes Virus Growth and Reactivation from Latency in Tonsils of Infected Calves
Authors: Perez Sandra , Inman Melissa , Doster Alan , Jones Clinton
Published: January 2005

Infection of calves with bovine herpesvirus 1 (BHV-1) results in transient immunosuppression that may lead to bacterium-induced pneumonia and, occasionally, death. Although sensory neurons in the trigeminal ganglia (TG) are the primary site of BHV-1 latency, viral genomes are detected in the tonsils of latently infected calves. Dexamethasone (DEX) consistently induces reactivation from latency, and viral gene expression is detected in TG and tonsils. In sensory neurons of latently infected calves, the latency-related (LR) gene is abundantly expressed and is required for reactivation from latency. In the present study, we compared the abilities of wild-type (wt) BHV-1 and a strain with a mutation in the LR gene (the LR mutant strain) to grow in the tonsils of infected calves and reactivate from latency. Lower levels of the LR mutant virus were detected in the tonsils of acutely infected calves. LR mutant viral DNA was consistently detected by PCR in the tonsils of latently infected calves, suggesting that the establishment of a latent or persistent infection occurred. Although the LR mutant did not reactivate from latency in vivo after DEX treatment, explantation of tonsil tissue from calves latently infected with the LR mutant yielded infectious virus. Relative to wt BHV-1, the LR mutant did not induce explant-induced reactivation as efficiently. These studies indicate that the LR gene promotes virus shedding from tonsil tissue during acute infection and reactivation from latency in tonsil tissue in vivo. We suggest that incorporation of the LR gene mutation into existing modified live vaccines would prevent reactivation from latency in neural and nonneural sites and would thus prevent transmission to other animals.
Multispacer Typing Technique for Sequence-Based Typing ofBartonella quintana
Authors: Foucault C. , La Scola B. , Lindroos H. , Andersson S. G. E. , Raoult D.
Published: January 2005

Bartonella quintanais a worldwide fastidious bacterium of the Alphaproteobacteria responsible for bacillary angiomatosis, trench fever, chronic lymphadenopathy, and culture-negative endocarditis. The recent genome sequencing of aB. quintanaisolate allowed us to propose a genome-wide sequence-based typing method. To ensure sequence discrimination based on highly polymorphic areas, we amplified and sequenced 34 spacers in a large collection ofB. quintanaisolates. Six of these exhibited polymorphisms and allowed the characterization of 4 genotypes. However, the strain variants suggested by the noncoding sequences did not correlate with the results of pulsed-field gel electrophoresis (PFGE), which suggested a higher degree of variability. Modification of the PFGE profile of one isolate after nine subcultures confirmed that rearrangement frequencies are high in this species, making PFGE unreliable for epidemiological purposes. The low extent of sequence heterogeneity in the species suggests a recent emergence of this bacterium as a human pathogen. Direct typing of natural samples allowed the identification of a fifth genotype in the DNA extracted from a human body louse collected in Burundi. We have named the typing technique herein described multispacer typing.
Detection of Human Influenza A Viruses by Loop-Mediated Isothermal Amplification
Authors: Poon Leo L. M. , Leung Cynthia S. W. , Chan Kwok H. , Lee Jack H. C. , Yuen Kwok Y. , Guan Yi , Peiris Joseph S. M.
Published: January 2005

Here we describe the use of the loop-mediated isothermal amplification (LAMP) method to detect human influenza viruses (H1 to H3). Our results were correlated 100% with results deduced from routine clinical diagnostic tests. In addition, we also developed a LAMP assay specific for human β-actin cDNA as a quality control test.
Effect of Inoculum Size on Detection of
Candida
Growth by the BACTEC 9240 Automated Blood Culture System Using Aerobic and Anaerobic Media
Authors: George Benjamin J. , Horvath Lynn L. , Hospenthal Duane R.
Published: January 2005

Simulated candidemia was produced with 20
Candida
isolates at three inoculum sizes (100, 10, and 1 CFU/ml of blood). Growth detection was better with larger inocula. The time to growth detection was shorter with larger inocula. Inoculum size does effect
Candida
growth detection and time to detection in BACTEC 9240 automated systems.
Cetyl-Pyridinium Chloride Is Useful for Isolation of
Mycobacterium tuberculosis
from Sputa Subjected to Long-Term Storage
Authors: Pardini Manuela , Varaine Francis , Iona Elisabetta , Arzumanian Erchanik , Checchi Francesco , Oggioni Marco Rinaldo , Orefici Graziella , Fattorini Lanfranco
Published: January 2005

Recovery of
Mycobacterium tuberculosis
from sputa treated with cetyl-pyridinium chloride (CPC) and stored for 20 ± 9 days was significantly higher than that from sputa that were untreated and processed by the
N
-acetyl-
l
-cisteine-NaOH method. Addition of CPC is useful for isolation of
M. tuberculosis
from sputa subjected to long-term storage received from remote areas of the world.
High Prevalence of Respiratory Viral Infections in Patients Hospitalized in an Intensive Care Unit for Acute Respiratory Infections as Detected by Nucleic Acid-Based Assays
Authors: Legoff Jérôme , Guérot Emmanuel , Ndjoyi-Mbiguino Angélique , Matta Mathieu , Si-Mohamed Ali , Gutmann Laurent , Fagon Jean-Yves , Bélec Laurent
Published: January 2005

Forty-seven bronchoalveolar lavages (BAL) were obtained from 41 patients with acute pneumonia attending an intensive care unit. By molecular diagnosis, 30% of total BAL and 63% of bacteria-negative BAL were positive for respiratory viruses. Molecular detection allows for high-rate detection of respiratory viral infections in adult patients suffering from severe pneumonia.
Activities of Available and Investigational Antifungal Agents against
Rhodotorula
Species
Authors: Diekema D. J. , Petroelje B. , Messer S. A. , Hollis R. J. , Pfaller M. A
Published: January 2005

Rhodotorula
species are emerging pathogens in immunocompromised patients. We report the in vitro activities of eight antifungals against 64
Rhodotorula
isolates collected in surveillance programs between 1987 and 2003.
Rhodotorula
strains are resistant in vitro to fluconazole (MIC at which 50% of the isolates tested are inhibited [MIC
50
], >128 μg/ml) and caspofungin (MIC
50
, >8 μg/ml). Amphotericin B (MIC
50
,1 μg/ml) and flucytosine (MIC
50
, 0.12 μg/ml) are both active in vitro, and the new and investigational triazoles all have some in vitro activity, with ravuconazole being the most active (MIC
50
, 0.25 μg/ml).
Polymorphism of the Human Immunodeficiency Virus Type 2 (HIV-2) Protease Gene and Selection of Drug Resistance Mutations in HIV-2-Infected Patients Treated with Protease Inhibitors
Authors: Damond F. , Brun-Vezinet F. , Matheron S. , Peytavin G. , Campa P. , Pueyo S. , Mammano F. , Lastere S. , Farfara I. , Simon F. , Chene G. , Descamps D.
Published: January 2005

We described the baseline polymorphism of the human immunodeficiency virus type 2 (HIV-2) protease gene from 94 treatment-naive patients and the longitudinal follow-up of 17 protease inhibitor-treated patients. Compared to the HIV-2 consensus sequences, baseline polymorphism involved 47 positions. Substitutions selected under treatment were observed at positions corresponding to HIV-1 resistance mutations as well as at positions of currently unknown impact on HIV-1.
Variable gp43 Secretion by
Paracoccidioides brasiliensis
Clones Obtained by Two Different Culture Methods
Authors: Berzaghi Rodrigo , Marques da Silva Silvia Helena , Pires de Camargo Zoilo
Published: January 2005

The main objectives of this study were to obtain clones of
Paracoccidioides brasiliensis
by two methods (micromanipulation and plating assay) and to determine if the secretion of the 43-kDa glycoprotein (gp43) is dependent on the clonal culture. The results show that the secretion of gp43 is not dependent on clonal cultures. Clones that originally were secretors of this molecule, after subculturing, lost this characteristic; on the other hand, clones that originally did not secrete gp43 began to secrete gp43 after subculturing.
Comparison of a New Commercial Test, GLABRATA RTT, with a Dipstick Test for Rapid Identification of
Candida glabrata
Authors: Willinger Birgit , Wein Susanne , Hirschl Alexander M. , Rotter Manfred L. , Manafi Mammad
Published: January 2005

This study compares the performance of a 3-h dipstick trehalose test with GLABRATA RTT, a new commercially available 20-min test for the rapid identification of
Candida glabrata
. With the exception of blood agar, GLABRATA RTT gave reliable results with all media tested and was always superior to the dipstick test.
Vibrio fluvialis
Peritonitis in a Patient Receiving Continuous Ambulatory Peritoneal Dialysis
Authors: Ratnaraja Natasha , Blackmore Timothy , Byrne Joan , Shi Shuning
Published: January 2005

We describe a case of peritonitis due to
Vibrio fluvialis
in a patient receiving continuous ambulatory peritoneal dialysis; we believe the case to be associated with the consumption of poorly prepared seafood. This was shown to be an important but rare cause of recurrent infection in our patient.
First Isolation of Metallo-β-Lactamase-Producing Multiresistant
Klebsiella pneumoniae
from a Patient in Brazil
Authors: Lincopan Nilton , McCulloch John Anthony , Reinert Cristina , Cassettari Valéria C. , Gales Ana C. , Mamizuka Elsa M.
Published: January 2005

A multiresistant
Klebsiella pneumoniae
isolate was taken from the blood of a 75-year-old patient with nosocomial pneumonia who developed septic shock and failed therapy with imipenem. The isolate presented an MIC of imipenem of 128 μg/ml, and the production of a metallo-β-lactamase was confirmed by phenotypic and genotypic techniques. We here report, for the first time, the detection of a metalloenzyme (IMP-1)-producing
K. pneumoniae
clinical strain in Latin America. The gene responsible for this phenotype was found to be
bla
IMP-1
, carried in a class 1 integron.
Molecular Epidemiology of Pneumococcal Colonization in Response to Pneumococcal Conjugate Vaccination in Children with Recurrent Acute Otitis Media
Authors: Bogaert D. , Veenhoven R. H. , Sluijter M. , Wannet W. J. W. , Rijkers G. T. , Mitchell T. J. , Clarke S. C. , Goessens W. H. F. , Schilder A. G. , Sanders E. A. M. , de Groot R. , Hermans P. W. M.
Published: January 2005

A randomized double-blind trial with a 7-valent pneumococcal conjugate vaccine was conducted in The Netherlands among 383 children, aged 1 to 7 years, with a history of recurrent acute otitis media. No effect of vaccination on the pneumococcal colonization rate was found. However, a shift in serotype distribution was clearly observed (R. Veenhoven et al., Lancet 361:2189-2195, 2003). We investigated the molecular epidemiology of 921 pneumococcal isolates retrieved from both the pneumococcal vaccine (PV) and control vaccine (CV) groups during the vaccination study. Within individuals a high turnover rate of pneumococcal restriction fragment end labeling genotypes, which was unaffected by vaccination, was observed. Comparison of the genetic structures before and after completion of the vaccination scheme revealed that, despite a shift in serotypes, there was clustering of 70% of the pneumococcal populations. The remaining isolates (30%) were equally observed in the PV and CV groups. In addition, the degree of genetic clustering was unaffected by vaccination. However, within the population genetic structure, nonvaccine serotype clusters with the serotypes 11, 15, and 23B became predominant over vaccine-type clusters after vaccination. Finally, overall pneumococcal resistance was low (14%), and, albeit not significant, a reduction in pneumococcal resistance as a result of pneumococcal vaccination was observed. Molecular surveillance of colonization in Dutch children shows no effect of pneumococcal conjugate vaccination on the degree of genetic clustering and the genetic structure of the pneumococcal population. However, within the genetic pneumococcal population structure, a clear shift toward nonvaccine serotype clusters was observed.