bluuebunny/crossref_metadata_2025 huggingface dataset:
The complex oral microflora of high‐risk individuals and groups and its role in the caries process
Authors: Beighton David
Published: July 2005

Abstract –  The involvement of the oral biofilm in the caries process requires re‐evaluation. The essential role of mutans streptococci (Streptococcus mutans and Streptococcus sobrinus) in the caries process is not proven. Acid production by dental plaque is not dependent upon the presence of mutans streptococci; caries occurs in the absence of these species and their presence does not necessarily indicate caries activity. Other oral bacteria, non‐mutans streptococci, Actinomyces spp. and Bifidobacterium spp., are acidogenic and aciduric. They outnumber mutans streptococci in dental plaque, and there are data which support a role for these bacteria in the initiation and progression of caries. Molecular studies demonstrate the great diversity and complexity of the flora associated with caries. Many taxa identified have not been cultured and the role of these taxa is not known. We have, in mutans streptococci, good markers of disease but not necessarily the aetiological agents of the disease. Considerably more research is required to investigate the transition of tooth surfaces from being intact and sound to the white spot lesion stage. A combination of conventional and molecular approaches are required to elucidate the involvement of an individual taxon and of microbial populations with particular traits in the caries process.
Cariogram – a multifactorial risk assessment model for a multifactorial disease
Authors: Bratthall Douglas , Hänsel Petersson Gunnel
Published: July 2005

Abstract –  This paper reviews some common methods for the assessment of caries risk. It also describes a new way of illustrating the caries risk profile of an individual, the Cariogram. Past caries experience and socioeconomic factors are often used for prediction of caries. As prediction models, the methods are simple, inexpensive and fast. However, they are not risk models, as they do not specify which particular risk factors are operating. Various biological factors can be used for risk assessment. Common ones are bacteria, diet and host factors. Taken separately, these biological factors often have limited predictive values. Socioeconomic factors often have a heavy impact on the biological factors as they can explain why an individual, for example, has a cariogenic diet or neglects oral hygiene. The biological factors are the immediate cause of the cavities. Caries experience is an illustration of how the host copes up with the biological activity. To facilitate the interpretation of biological data, the Cariogram was developed. It is a computer program showing a graphical picture that illustrates a possible overall caries risk scenario. The program contains an algorithm that presents a ‘weighted’ analysis of the input data, mainly biological factors. It expresses as to what extent different etiological factors of caries affect caries risk. The Cariogram identifies the caries risk factors for the individual and provides examples of preventive and treatment strategies to the clinician.
Sociobehavioural risk factors in dental caries – international perspectives
Authors: Petersen Poul Erik
Published: July 2005

Abstract –  Diseases probably have their roots in a complex chain of environmental and behavioural events which are shaped by broader socioeconomic determinants. Most studies of sociobehavioural risk factors in dental caries have been carried out in industrialized countries, but such reports from low‐ and middle‐income countries have been published in recent years. World Health Organization international collaborative studies and other international studies of social factors in dental caries using the same methodology provide empirical evidence of social inequality in oral health across countries and across oral health care systems. The paper highlights the challenges to dental public health practice, particularly the importance of risk assessment in estimating the potential for prevention. In future public health programmes, systematic risk factor assessment may therefore be instrumental in the planning and surveillance of oral health promotion and oral disease intervention programmes.
Deoxycoformycin (pentostatin) in the treatment of splenic marginal zone lymphoma (SMZL) with or without villous lymphocytes
Authors: Iannitto Emilio , Minardi Viviana , Calvaruso Giuseppina , Ammatuna Emanuele , Florena Ada Maria , Mulè Antonino , Tripodo Claudio , Quintini Gerlando , Abbadessa Vincenzo
Published: July 2005

Abstract:  Background: Splenic marginal zone lymphoma (SMZL) is an infrequent B‐cell neoplasm that pursues an indolent course. Signs and symptoms, mostly related to hypersplenism, are successfully managed by splenectomy. However, the therapy of patients who are not fit for a surgical procedure or who relapse after splenectomy, is still an unsettled issue. Patients and methods: We report a phase‐II study on 16 patients with SMZL, three therapy naïve and 13 pretreated, all showing systemic symptoms or progressive worsening of peripheral cytopenia, who were treated with pentostatin at a dose of 4 mg/m2 every other week for 6–10 wk. In relapsed patients, the median interval between diagnosis and treatment was 26 month (range: 8–49). Results: Overall, 68% of the patients showed a clinical response. Two out three patients, who received pentostatin as first line therapy, attained a complete response (CR). One CR and seven minor or good haematological responses were recorded in relapsed patients. Treatment toxicity, mostly haematological, proved manageable. With a median follow‐up of 35 month the median overall survival (OS) is 40 month and the median progression free survival (PFS) is 18 month. Conclusion: Our data show that pentostatin administered every other week has a good degree of activity in the treatment of SMZL and suggest that this schedule could be considered a possible therapeutic option for patients who are not fit for splenectomy or have relapsed.
Self‐reported quality of life in elderly patients with aggressive non‐Hodgkin's lymphoma treated with CHOP chemotherapy
Authors: Doorduijn Jeanette , Buijt Ivonne , van der Holt Bronno , Steijaert Monique , Uyl‐de Groot Carin , Sonneveld Pieter
Published: July 2005

Abstract:  We studied the impact of CHOP chemotherapy on the quality of life (QoL) of elderly patients with aggressive non‐Hodgkin's lymphoma (NHL). 132 patients aged 65 or older, who participated in a randomized, multicenter trial, completed QoL questionnaires (EuroQol‐5D, EORTC QLQ‐C30 and MFI‐20) on 8 predefined time‐points before, during and following treatment. At baseline, QoL was significantly better on almost all dimensions in patients with a lower compared to patients with a higher age‐adjusted International Prognostic Index (aaPI). During treatment, physical and role functioning and global QoL deteriorated and fatigue increased in the lower aaPI group, whereas QoL of the higher aaPI group remained stable. During follow‐up, the QoL was significantly better for patients in complete response (CR) or partial remission (PR) than for patients with progression/relapse. Soon after completion of therapy, the QoL of the lower aaPI group returned to pretreatment levels or better, while patients with higher aaPI showed a significant improvement in QoL compared to baseline levels. The effect of CHOP on the quality of life of elderly patients could be used in counseling this group of patients.
Macrocytosis associated with monoclonal gammopathy
Authors: Horstman Andrew L. , Serck Stephanie L. , Go Ronald S.
Published: July 2005

Abstract:  Objective: To report the potential association between unexplained macrocytosis and monoclonal gammopathy. Methods: We retrospectively reviewed the medical records of patients who had monoclonal protein detected by serum electrophoresis and immunofixation from October 1999 until September 2003 at our institution. Patients with concomitant macrocytosis were included in this study. We collected data on patient demographics, evaluations performed for macrocytosis, pertinent laboratory tests relevant to the diagnosis of monoclonal gammopathy and presence of associated hematologic disorders. Results: We identified 258 patients with monoclonal gammopathy. Thirty‐one (12%) of them had concomitant macrocytosis. Of the latter group, 14 (5%) patients had no identifiable cause of macrocytosis after thorough evaluation and were considered to have macrocytosis associated with monoclonal gammopathy. The median values for mean cell volume and serum monoclonal protein were 103.9 fL (range 100.8–109.8) and 1.95 gm/dL (range 0.8–4.3), respectively. Most patients had IgG (71%) and kappa light chain (79%). All of the 11 (of 14) patients who underwent a bone marrow biopsy as part of the initial evaluation had megaloblastoid maturation of the erythroid precursors. No correlation was found between the level of serum monoclonal protein and the degree of macrocytosis (r = +0.48, P = 0.08). After a median follow‐up of 22.5 month (range 3–60+), all but one patient had persistent but stable macrocytosis. Conclusion: Macrocytosis can be a manifestation of monoclonal gammopathy. Disorders associated with monoclonal gammopathy should be considered in the differential diagnoses during evaluation of macrocytosis.
Expression of cyclin A1 and cell cycle proteins in hematopoietic cells and acute myeloid leukemia and links to patient outcome
Authors: Ekberg Jenny , Holm Caroline , Jalili Sara , Richter Johan , Anagnostaki Lola , Landberg Göran , Persson Jenny Liao
Published: July 2005

Abstract:  Abnormal expression of several key regulators essential for G1/S transitions has been implicated in tumorigenesis. A critical role of cyclin A1 in the development of acute myeloid leukemia (AML) has previously been demonstrated in transgenic mice. Our present study focused on the expression and prognostic significance of cyclin A1 and a panel of cell cycle regulatory proteins including cyclin A2, cyclin B1, cyclin E, CDK1, CDK2, p21 and p27 in bone marrow samples from 40 patients with AML. Freshly isolated CD34+ hematopoietic cells and bone marrow samples from 10 healthy donors were also assessed for cell type‐ and subcellular‐specific expression of the cell cycle regulatory proteins. The level of cyclin A1 expression was the only factor that showed a significant correlation with patient outcome. In log‐rank test stratified by levels of cyclin A1 expression, patients with high levels of cyclin A1 had significantly worse overall survival (OS) (P = 0.012) compared to those with low levels. Further, patients with high levels of cyclin A1 had significantly lower disease‐free survival (DFS) (P = 0.028). Multivariate analysis indicated that cyclin A1 protein expression was an independent prognostic factor for predicting DFS (P = 0.035) and OS (P = 0.045). No correlation between cyclin A1 expression and age was found. However, expression of cyclin A2, cyclin B1, cyclin E, CDK1, CDK2, p21 and p27 did not show prognostic significance in these AML patients.
The influence of uridine diphosphate glucuronosyl transferase 1A promoter polymorphisms, βS‐globin gene haplotype, co‐inherited α‐thalassemia trait and Hb F on steady‐state serum bilirubin levels in sickle cell anemia
Authors: Adekile A. , Kutlar F. , McKie K. , Addington A. , Elam D. , Holley L. , Clair B. , Kutlar A.
Published: July 2005

Abstract:  Purpose: Homozygosity for the (AT)7 allele of uridine diphosphate glucuronosyl transferase 1A (UGT1A1) gene polymorphism is associated with increased bilirubin levels in sickle cell anemia (SCA). In the present study, in addition to UGT1A1 promoter genotype, serum bilirubin level was related to other genetic modifiers –βS‐globin gene haplotype, Hb F, co‐inherited α‐thal trait, age and gender. Methods: The patients were randomly selected from the sickle cell clinic, Medical College of Georgia. UGT1A1 promoter polymorphisms were determined using automated sequencing. Other investigations were with standard techniques. Results: There were 67 SCA patients (41 males and 26 females), aged 2–44 yr (mean of 20.6 ± 10.7). Ten (14.9%) patients were homozygous for the (AT)6 UGT1A1 allele, 35 (52.2%) were heterozygous for (AT)6 and (AT)7 alleles while 22 (32.8%) were homozygous for (AT)7. Serum bilirubin was significantly higher in the homozygous (AT)7 group (3.7 ± 1.5, 3.8 ± 2.3 and 5.6 ± 2.4 mg/dL, respectively). It was also significantly higher in males than females and in patients aged >10 yr. There was a significant negative linear correlation (r = −0.304, P = 0.016) of serum bilirubin with Hb F. The β‐globin haplotype and co‐existing α‐thal trait did not have any significant influence on serum bilirubin levels. Patients on hydroxyurea were older, had lower Hb F, but higher mean serum bilirubin. The latter also was signifcantly higher among those with UGT1A1 (AT)7 homozygosity. Conclusions: Apart from UGT1A1 (AT)7 homozygosity, Hb F, age and gender are the other factors that significantly influence serum bilirubin level in SCA.
Combination chemotherapy with gemcitabine and vinorelbine in the treatment of relapsed or refractory diffuse large B‐cell lymphoma: a phase‐II trial by the Hellenic Cooperative Oncology Group
Authors: Papageorgiou Efstathios S. , Tsirigotis Panagiotis , Dimopoulos Meletios , Pavlidis Nikolaos , Fountzilas George , Papageorgiou Sotirios , Economopoulos Theofanis
Published: July 2005

Abstract:  To investigate the efficacy and toxicity of the combination of gemcitabine and vinorelbine in patients with relapsed or refractory diffuse large B‐cell lymphoma (DLBL), 22 patients with relapsed or refractory DLBL were treated with gemcitabine 1000 mg/m2 and vinorelbine 30 mg/m2 on days 1 and 8 every 3 wk for a maximum of six cycles. Fourteen patients were considered chemosensitive while eight patients were considered chemoresistant to the last treatment regimen. All 22 patients were assessed for response to treatment. Three patients (14%) achieved complete remission and eight patients (36%) had partial remission of their disease, with an overall response rate of 50%. With a median follow up of 44 months, the median time to progression (TTP) for all patients was 8.1 months while the median overall survival (OS) was 12.9 months. Toxicity was minimal and all patients were treated on an outpatient basis. The combination of gemcitabine and vinorelbine is an effective and well‐tolerated regimen for patients with relapsed of refractory DLBL.
Long‐term experience with low‐dose interferon‐α and PUVA in the management of early mycosis fungoides
Authors: Rupoli Serena , Goteri Gaia , Pulini Stefano , Filosa Alessandra , Tassetti Angela , Offidani Massimo , Filosa Giorgio , Mozzicafreddo Giorgio , Giacchetti Alfredo , Brandozzi Giuliano , Cataldi Ivana , Barulli Sara , Ranaldi Renzo , Scortechini Anna Rita , Capretti Roberta , Fabris Guidalberto , Leoni Pietro ,   (for the Marche Regional Multicentric Study Group of Cutaneous Lymphomas)
Published: July 2005

Abstract:  Objectives: Combined high‐dose Interferon‐α and psoralen plus ultraviolet A irradiation (PUVA) have been reported to be effective in the treatment of early mycosis fungoides (MF); however, our study is the first controlled prospective study in the literature exploring the activity and tolerability of the combination with low dosages and evaluating further clinical outcome of early‐MF patients. Methods: We carried out a multicentric prospective Phase II clinical study on 89 patients with early‐stage IA to IIA MF treated for 14 months with low‐dose IFN‐α2b (6–18 MU/wk) and PUVA. Treatment success was analysed in terms of freedom from treatment failure. Results and conclusions: Complete remission (CR) was achieved in 84% and an overall response rate in 98% of cases: six‐month CR was associated with a non‐confluent skin infiltrate at histology (P = 0.044) and 14‐month CR with high epidermal CD1a+ dendritic‐cell density (P = 0.030). The combination protocol was successfully tolerated and the most common reason of ‘failure’ was related to relapse and not to toxicity. Sustained remissions were achieved in 20% of patients. High CD8+ lymphoid T‐cell density was associated with a lower relapse rate (P = 0.002). We think that our combination therapy can be considered an alternative approach compared with other modalities. Good immunological host surveillance in the skin lesions seems to be an optimal basis for the therapeutic success.
Etiology of thrombocytopenia in all patients treated with heparin products
Authors: Laber Damian A. , Martin Monte E.
Published: July 2005

Abstract:  Purpose: To characterize the cause of thrombocytopenia in all patients treated with heparin products, to determine the incidence of heparin‐induced thrombocytopenia (HIT) in unselected hospitalized patients, and to have modern data of the magnitude of this problem. Methods: Retrospective hospital‐based cohort study. During a random 2‐month period, we reviewed the medical records of all patients treated with heparin agents, screened them for thrombocytopenia, and determined the cause of it. Results: Out of 674 patients who received heparin products, 110 (16%) had thrombocytopenia. The most common causes included cancer chemotherapy, surgery, sepsis, and medications. Three patients met the clinical criteria for HIT. One had antibodies for heparin‐platelet factor‐4, and received a direct thrombin inhibitor. The other two individuals had a clinical syndrome that resembled immune HIT, but were not tested for HIT antibodies. One suffered a thrombotic episode that led to the death of her fetus. The other died of a possible thromboembolism. Conclusions: This study provides evidence‐based data for the differential diagnosis of thrombocytopenia after treatment with heparin products. Our findings suggest that increased awareness of the HIT syndrome might reduce morbidity and mortality. Patients exposed to heparin products, who develop thrombocytopenia, should not be overlooked.
Adult colonic intussusception
Authors: Lai Eric Chun‐Hung , Lee Janet Fung‐Yee , Ng Simon Siu‐Man , Yiu Raymond Ying‐Chang , Leung Ka‐Lau
Published: July 2005

Objective:  To study the clinical characteristics, investigations, management and pathology in adult colonic intussusception.

Methods:  A retrospective study of seven adult patients who were operated on for the diagnosis of colonic intussusception in our department from January 1999 to September 2003 was carried out. The case records of the seven patients were retrieved. Their clinical presentations, investigations, operative records and pathology were studied.

Results:  The mean age of the patients was 67.4 years. There were four men and three women. Five of the patients presented with chronic non‐specific symptoms and the other two patients presented with acute symptoms. Transabdominal ultrasonography (USG) and computed tomography (CT) scan were used for investigation. In the majority of the patients (87.5%), radiological features of intussusception were shown by USG or CT scan. All seven patients underwent laparotomy exploration with bowel resection. Six patients (85.7%) had lead points with definable pathology. Within the lead points, five patients had primary malignant lesions.

Conclusion:  With more liberal use of imaging techniques, more adult intussusception can be diagnosed before explorative surgery. Ultrasound would be an ideal tool for initial evaluation. In view of the high chance of associated malignancy, adult colonic intussusception should be resected without prior reduction at the time of surgery.
Aortoesophageal fistula: A rare and dreaded cause of gastrointestinal haemorrhage
Authors: Kapoor Sorabh , Singh Rajneesh K. , Chattopadhyay Tushar K.
Published: July 2005

Aortoesophageal fistula (AEF) is an uncommon cause of gastrointestinal bleeding. Most cases are identified post‐mortem after an exsanguinating haemorrhage. Common causes include rupture of thoracic aortic aneurysm, oesophageal foreign bodies and thoracic surgery. AEF following previous aortic or oesophageal surgery are termed as secondary AEF, whereas fistulas that are the result of a ruptured aortic aneurysm, aortic dissection, foreign bodies, oesophageal cancer etc. are termed as primary AEF. The presentation is with the classic Chiari's triad, however, not all components might be present. Endoscopy, computed tomography scan and aortography are useful for diagnosis and for identifying the site of the fistula. Emergency surgery is the treatment of choice, although isolated reports of aortic stenting are available. Aortic repair, restoration of circulation and delayed oesophageal reconstruction are essential principles in the management of aortoesophageal fistula. With early recognition and proper management, more survivors are being reported in literature.
Gallstone ileus masquerading as an obstructed femoral hernia
Authors: Saad Al Skaini Mohammad , Ezzedien Rabie Mohammad , Al Ghamdi Abdullah H. , Kandeel Ahmad Y , Sulaiman Al Mahdi Mohammad
Published: July 2005

Gallstone ileus affects primarily elderly women. In many instances, no history of previous biliary disease is discernible. Although it has been known since the 17th century, gallstone ileus continues to present in such an intriguing way that, more often than not, the diagnosis is made intraoperatively. A 68‐year‐old woman, presented with abdominal pain and vomiting. She had a slightly distended and mildly tender abdomen, and the bowel sounds were normal. There was an irreducible, slightly tender right femoral hernia. Plain X‐rays of the abdomen were normal and an ultrasound showed a contracted fibrotic gall bladder with small stones. Laparoscopy failed to detect an obstructing gallstone, which was discovered in a second laparotomy. Cholecystectomy and fistula closure were carried out in the same sitting. A retrospective review of a preoperative Gastrografin study showed clear radiological evidence of the condition. Gallstone ileus tends to be forgotten as a cause of bowel obstruction or abdominal pain in elderly women. With this in mind, careful interpretation of a Gastrografin study might provide the diagnosis preoperatively. In relatively fit patients, cholecystectomy and fistula closure could be safely added to the emergency enterolithotomy.
Laparoscopy for diagnosis and staging of hepatobiliary malignancies
Authors: Abdalla Eddie K. , Aloia Thomas A. , Vauthey Jean‐Nicolas
Published: July 2005

Objective:  To review the role of diagnostic laparoscopy (DL) for staging of malignant diseases of the liver and biliary tract.

Methodology:  Critical review of the current literature.

Results:  Analysis of the utility of DL in hepatobiliary cancers depends on several criteria, particularly in the era of high quality prelaparotomy and pre‐DL imaging. Selection criteria for DL, selection criteria for resection, definition of resectability, patterns of intra‐ and extrahepatic spread, association with underlying liver disease and frequency of indications for palliative laparotomy impact the utility of DL depending on the disease studied.

Conclusions:  DL has a very limited role for staging patients with colorectal liver metastases as a result of expanding definitions of resectability, multistage approaches to bilateral metastases, and methods to increase resectability such as portal vein embolization and preoperative chemotherapy. For hepatocellular carcinoma, DL can be useful for staging patients with advanced tumours and cirrhosis, and might have an emerging role for the evaluation of transplant candidates with equivocal imaging findings. For biliary cancers, DL is indicated for patients with advanced stage hilar cholangiocarcinoma and gall bladder carcinoma.
Effect of pH and Antibiotics on Microbial Overgrowth in the Stomachs and Duodena of Patients Undergoing Percutaneous Endoscopic Gastrostomy Feeding
Authors: O'May Graeme A. , Reynolds Nigel , Smith Aileen R. , Kennedy Aileen , Macfarlane George T.
Published: July 2005

Enteral nutrition via a percutaneous endoscopic gastrostomy (PEG) tube is often part of management in patients with dysphagia due to neurological or oropharyngeal disease. Gastrostomy placement can affect normal innate defense mechanisms in the upper gut, resulting in bacterial overgrowth. In this study microbiological investigations were done with gastric and duodenal aspirates from 20 patients undergoing PEG tube placement and PEG tubes from 10 patients undergoing tube replacement. Aspirate and PEG tube microbiotas were assessed by using viable counts and selective solid media followed by aerobic and anaerobic incubation to assess cell viabilities. The antibiotic susceptibility profiles of the isolates were determined by the disk diffusion method, and gas chromatography was used to study the bacterial metabolic products in the aspirates. The aspirates and PEG tubes contained mainly streptococci, staphylococci, lactobacilli, yeasts, and enterobacteria. Enterococci were detected only in PEG tube biofilms and not in aspirates. Gastric pH affected the composition of the aspirate microbiotas but not the total microbial counts. Staphylococci,
Escherichia coli
, and
Candida
spp. were isolated only from antibiotic-treated patients, despite the sensitivities of the bacteria to the agents used. Antibiotic treatment had no effect on the incidence of infection or the length of hospital stay in these patients.
Diagnostic Value of Measuring Epstein-Barr Virus (EBV) DNA Load and Carcinoma-Specific Viral mRNA in Relation to Anti-EBV Immunoglobulin A (IgA) and IgG Antibody Levels in Blood of Nasopharyngeal Carcinoma Patients from Indonesia
Authors: Stevens Servi J. C. , Verkuijlen Sandra A. W. M. , Hariwiyanto Bambang , Harijadi  , Fachiroh Jajah , Paramita Dewi K. , Tan I. Bing , Haryana Sophia M. , Middeldorp Jaap M.
Published: July 2005

Nasopharyngeal carcinoma (NPC) is a prevalent malignancy in Southeast Asia and is strongly associated with Epstein-Barr virus (EBV). We investigated the primary diagnostic value of circulating EBV DNA and anti-EBV immunoglobulin G (IgG) and IgA levels in Indonesian NPC patients (
n
= 149). By a 213-bp Epstein-Barr virus nuclear antigen 1 (EBNA1)-based real-time LightCycler PCR, 72.5% of patients were positive for EBV DNA in whole blood, with 29.5% having levels above a previously determined clinical cutoff value (COV) of 2,000 EBV DNA copies/ml, the upper level in healthy carriers. In a 99-bp LightCycler PCR, 85.9% of patients were positive and 60.4% had levels above the COV. This assay quantified a significantly higher EBV load than the 213-bp PCR assay (
P
< 0.0001), suggesting that circulating EBV DNA is fragmented. Using data from 11 different studies, we showed a significant inverse correlation between PCR amplicon size and the percentage of patients positive for circulating EBV DNA (Spearman's rho = −0.91;
P
< 0.0001). EBV DNA loads were unrelated to anti-EBV IgG or IgA levels, as measured by VCA-p18 and EBNA1-specific synthetic peptide-based enzyme-linked immunosorbent assays. The presence of circulating tumor cells was assessed by amplification of BamHI-A rightward frame 1 (BARF1) mRNA, a viral oncogene abundantly expressed in EBV-carrying carcinomas but virtually absent from EBV-associated lymphomas. Despite high EBV DNA loads and the presence of EBNA1 and human U1A small nuclear ribonucleoprotein mRNA, BARF1 mRNA was never detected in blood. We conclude that amplicon size significantly influences EBV DNA load measurement in NPC patients. The circulating EBV DNA load is independent of serological parameters and does not reflect intact tumor cells. The primary diagnostic value of the EBV DNA load for the detection of NPC is limited.
Detection and Typing of Integrons in Epidemic Strains of
Acinetobacter baumannii
Found in the United Kingdom
Authors: Turton Jane F. , Kaufmann Mary E. , Glover Judith , Coelho Juliana M. , Warner Marina , Pike Rachel , Pitt Tyrone L.
Published: July 2005

Integrons were sought in
Acinetobacter
isolates from hospitals in the United Kingdom by integrase gene PCR. Isolates were compared by pulsed-field gel electrophoresis, and most belonged to a small number of outbreak strains or clones of
A. baumannii
, which are highly successful in the United Kingdom. Class 1 integrons were found in all of the outbreak isolates but in none of the sporadic isolates. No class 2 integrons were found. Three integrons were identified among the main outbreak strains and clones. While a particular integron was usually associated with a strain or clone, some members carried a different integron. Some integrons were associated with more than one strain. The cassette arrays of two of the integrons were very similar, both containing gene
aacC1
, which confers resistance to gentamicin, two open reading frames coding for unknown products (orfX, orfX′), and gene
aadA1a
, which confers resistance to spectinomycin and streptomycin. The larger of these integrons had two copies of the first (orfX) of the gene cassettes coding for unknown products. The third integron, with a cassette array containing gene
aacA4
, which codes for amikacin, netilmicin, and tobramycin resistance; a chloramphenicol acetyltransferase,
catB8
; and gene
aadA1
, conferring resistance to spectinomycin and streptomycin, was associated with an OXA-23 carbapenemase-producing clone, which has spread rapidly in hospitals in the United Kingdom during 2003 and 2004. These integron cassette arrays have been found in other outbreak strains of
A. baumannii
from other countries. We conclude that integrons are useful markers for epidemic strains of
A. baumannii
and that integron typing provides valuable information for epidemiological studies.
Genomic and Bioinformatics Analyses of HAdV-4vac and HAdV-7vac, Two Human Adenovirus (HAdV) Strains That Constituted Original Prophylaxis against HAdV-Related Acute Respiratory Disease, a Reemerging Epidemic Disease
Authors: Purkayastha Anjan , Su Jing , McGraw John , Ditty Susan E. , Hadfield Ted L. , Seto Jason , Russell Kevin L. , Tibbetts Clark , Seto Donald
Published: July 2005

Vaccine strains of human adenovirus serotypes 4 and 7 (HAdV-4vac and HAdV-7vac) have been used successfully to prevent adenovirus-related acute respiratory disease outbreaks. The genomes of these two vaccine strains have been sequenced, annotated, and compared with their prototype equivalents with the goals of understanding their genomes for molecular diagnostics applications, vaccine redevelopment, and HAdV pathoepidemiology. These reference genomes are archived in GenBank as HAdV-4vac (35,994 bp;
AY594254
) and HAdV-7vac (35,240 bp; AY594256). Bioinformatics and comparative whole-genome analyses with their recently reported and archived prototype genomes reveal six mismatches and four insertions-deletions (indels) between the HAdV-4 prototype and vaccine strains, in contrast to the 611 mismatches and 130 indels between the HAdV-7 prototype and vaccine strains. Annotation reveals that the HAdV-4vac and HAdV-7vac genomes contain 51 and 50 coding units, respectively. Neither vaccine strain appears to be attenuated for virulence based on bioinformatics analyses. There is evidence of genome recombination, as the inverted terminal repeat of HAdV-4vac is initially identical to that of species C whereas the prototype is identical to species B1. These vaccine reference sequences yield unique genome signatures for molecular diagnostics. As a molecular forensics application, these references identify the circulating and problematic 1950s era field strains as the original HAdV-4 prototype and the Greider prototype, from which the vaccines are derived. Thus, they are useful for genomic comparisons to current epidemic and reemerging field strains, as well as leading to an understanding of pathoepidemiology among the human adenoviruses.
Variations inemmType among Group A Streptococcal Isolates Causing Invasive or Noninvasive Infections in a Nationwide Study
Authors: Ekelund Kim , Darenberg Jessica , Norrby-Teglund Anna , Hoffmann Steen , Bang Didi , Skinhøj Peter , Konradsen Helle Bossen
Published: July 2005

Since the late 1980s several studies have described the increased incidence and severity of invasive group A streptococcal (GAS) infections. However, most studies on GAS pathogenesis have focused on information obtained during outbreaks. We analyzed isolate distribution and host susceptibility as part of a nationwide prospective surveillance study performed between January 2001 and August 2002. GAS isolates collected from 201 patients with invasive infections, 335 patients with noninvasive infections, and 17 asymptomatic carriers were characterized with respect to theiremmtypes and superantigen genotypes. The superantigen-neutralizing capacity and levels of antibodies against streptolysin O and DNAse B were determined for isolates from the sera from 36 invasive cases and 91 noninvasive cases.emmtype 1 (emm-1) isolates were significantly more common among invasive cases, whereasemm-4,emm-6, andemm-12 dominated among the noninvasive cases. The distributions of the phage-associated superantigen genes (speA,speC,speH,speI,ssa) differed among invasive and noninvasive isolates, mainly due to their linkage to certainemmtypes. No significant differences in serum superantigen-neutralizing capacities were observed. The levels of anti-streptolysin O and anti-DNAse B antibodies were highest in the sera from invasive cases. Our study emphasizes the importance of obtaining data during years with stable incidences, which will enable evaluation of future outbreak data.
AmpC Disk Test for Detection of Plasmid-Mediated AmpC β-Lactamases in
Enterobacteriaceae
Lacking Chromosomal AmpC β-Lactamases
Authors: Black Jennifer A. , Moland Ellen Smith , Thomson Kenneth S.
Published: July 2005

Although plasmid-mediated AmpC β-lactamases were first reported in the late 1980s, many infectious disease personnel remain unaware of their clinical importance. These enzymes are typically produced by isolates of
Escherichia coli
,
Klebsiella
spp.,
Proteus mirabilis
, and
Salmonella
spp. and are associated with multiple antibiotic resistance that leaves few therapeutic options. Plasmid-mediated AmpC β-lactamases have been associated with false in vitro susceptibility to cephalosporins. Many laboratories do not test for this resistance mechanism because current tests are inconvenient, subjective, lack sensitivity and/or specificity, or require reagents that are not readily available. In this study a new test, the AmpC disk test, based on filter paper disks impregnated with EDTA, was found to be a highly sensitive, specific, and convenient means of detection of plasmid-mediated AmpC β-lactamases in organisms lacking a chromosomally mediated AmpC β-lactamase. Using cefoxitin insusceptibility as a screen, the test accurately distinguished AmpC and extended-spectrum β-lactamase production and differentiated AmpCs from non-β-lactamase mechanisms of cefoxitin insusceptibility, such as reduced outer membrane permeability. The test is a potentially useful diagnostic tool. It can provide important infection control information and help to ensure that infected patients receive appropriate antibiotic therapy.
Drug Resistance Evolution of a
Mycobacterium tuberculosis
Strain from a Noncompliant Patient
Authors: Meacci Francesca , Orrù Germano , Iona Elisabetta , Giannoni Federico , Piersimoni Claudio , Pozzi Gianni , Fattorini Lanfranco , Oggioni Marco R.
Published: July 2005

The emergence and spread of multidrug-resistant (MDR)
Mycobacterium tuberculosis
(MT) represents a worldwide health care problem because of the difficulty in treating these infections. Development of drug resistance in MT arises mainly by mutation of chromosomal genes. To investigate the evolution of a MT population during a long-lasting infection, the phenotypic and genotypic changes in the drug resistance of 10 sequential MT isolates from a noncompliant chronically infected patient were investigated. During more than 12 years of active disease, a MDR population developed; molecular typing showed one single parental strain that infected the patient and persisted throughout the disease. Molecular analysis of the drug resistance-related genes revealed that discrete subpopulations evolved over time from the parental strain by acquiring and accumulating resistance-conferring mutations to isoniazid, rifampin, and streptomycin. Overall, these observations indicate that during a chronic infection, several subpopulations may coexist in the same patient with different drug susceptibility profiles.
Genetic Relatedness between Pneumococcal Populations Originating from the Nasopharynx, Adenoid, and Tympanic Cavity of Children with Otitis Media
Authors: Tonnaer Edith L. , Rijkers Ger T. , Meis Jacques F. , Klaassen Corné H. , Bogaert Debby , Hermans Peter W. , Curfs Jo H.
Published: July 2005

Previous studies have shown that
Streptococcus pneumoniae
exists in both middle ear effusions and the upper respiratory region from children with otitis media with effusion (OME), but it remains unclear whether these strains represent genetically identical clones. Therefore, it cannot be determined whether these bacteria originate from a common source. To determine the presence of pneumococci at different anatomical locations of OME patients, conventional culture and PCR techniques were used. To analyze the possible genetic relatedness between pneumococci from different anatomical sites, molecular typing by amplified fragment length polymorphism was utilized. The percentage of middle ear effusions of OME patients that are positive for pneumococci after PCR analysis (13%) was higher than after conventional culture (5%). Molecular fingerprints from pneumococci derived from two different anatomic sites within patients were very similar in 80% of OME patients and in 90% of acute otitis medium patients, indicating their genetic relatedness. Biofilm formation or pneumococcal L-forms probably play a role in OME, since culture-negative effusions prove to contain pneumococcal DNA. Bacteria involved in this process most likely originate from the nasopharynx since they show a close genetic relatedness with their nasopharyngeal counterparts.
Drug Susceptibility Testing of
Mycobacterium tuberculosis
by a Nitrate Reductase Assay Applied Directly on Microscopy-Positive Sputum Samples
Authors: Musa Humberto R. , Ambroggi Marta , Souto Alejandro , Ängeby K. A. Kristian
Published: July 2005

Current methods for drug susceptibility testing of
Mycobacterium tuberculosis
are either costly or slow. As the prevalence of multidrug-resistant strains increases, the need for fast, reliable, and inexpensive methods that can also be applied in settings with scarce resources is obvious. We evaluated a rapid colorimetric nitrate reductase assay (NRA) for direct drug susceptibility testing of
M. tuberculosis
directly from clinical sputum samples with positive microscopy results for acid-fast bacilli with more than 10 acid-fast bacilli per high-power field. We have saved valuable time by omitting the preisolation step. The sensitivity (ability to detect true drug resistance) and specificity (ability to detect true drug susceptibility) of the direct NRA, using the direct proportion method as the reference, were 100 and 100%, 93 and 100%, 76 and 100%, and 55 and 99% for rifampin, isoniazid, streptomycin, and ethambutol, respectively, when tested on
M. tuberculosis
strains present in 121 samples. The results were in most cases available in 14 days. The direct NRA could be used as a rapid, inexpensive, and accurate method to determine rifampin and isoniazid susceptibility directly from sputum. The technique might become a valid alternative to traditional methods, especially in low-income countries.
Susceptibility of
Neisseria meningitidis
to 16 Antimicrobial Agents and Characterization of Resistance Mechanisms Affecting Some Agents
Authors: Jorgensen James H. , Crawford Sharon A. , Fiebelkorn Kristin R.
Published: July 2005

Neisseria meningitidis
represents a pathogen of great public health importance in both developed and developing countries. Resistance to some antimicrobial agents used either for therapy of invasive infections or for prophylaxis of case contacts has long been recognized, although specific guidelines for susceptibility testing have not been fully developed. We have examined the susceptibilities of a collection of 442 meningococcal clinical isolates from 15 countries to 16 antimicrobial agents. These included isolates recovered between 1917 and 2004, with representatives of all major serogroups. All isolates were tested by the Clinical and Laboratory Standards Institute (formerly NCCLS) broth microdilution method using Mueller-Hinton lysed horse blood broth, while a subset of 102 isolates was tested by agar dilution using Mueller-Hinton sheep blood agar. Most isolates provided adequate growth for MIC determinations by both broth and agar methods. Growth in broth was enhanced by CO
2
incubation and was required for two strains (1.7%). MICs of the study drugs compared favorably between the broth and agar methods (79 to 100% essential agreement), and MICs also generally agreed closely (92 to 100% essential agreement, excluding azithromycin) between broth tests incubated in the two different atmospheres. Elevated penicillin and ampicillin MICs (≥0.12 μg/ml and ≥0.25 μg/ml, respectively) occurred in 14.3% and 8.6% of strains and were associated with polymorphisms of the
penA
gene encoding a modified penicillin-binding protein 2. None of the 442 isolates produced beta-lactamase. Elevated tetracycline and doxycycline (but not minocycline) MICs were associated with efflux-mediated resistance encoded by
tet
(B) in 13 strains. Resistance to sulfisoxazole in 21.7% of strains and to trimethoprim-sulfamethoxazole in 21.0% resulted from polymorphisms of
folP
encoding a modified dihydropteroate synthetase. Seven strains were resistant to rifampin due to mutations in the
rpoB
gene, and two strains were resistant to chloramphenicol due to production of chloramphenicol acetyltransferase mediated by
catP
. Two strains had reduced quinolone susceptibility due to mutations of
gyrA
. The determination of the susceptibilities of a large group of meningococcal strains (including strains with characterized resistance mechanisms) to 16 antimicrobial agents has served as the essential first step in defining susceptibility testing breakpoints specific for this organism.
Detection of
Balamuthia
Mitochondrial 16S rRNA Gene DNA in Clinical Specimens by PCR
Authors: Yagi Shigeo , Booton Gregory C. , Visvesvara Govinda S. , Schuster Frederick L.
Published: July 2005

Balamuthia mandrillaris
is a free-living ameba that causes granulomatous amebic encephalitis in both immunocompromised and immunocompetent individuals. Because of a lack of pathognomonic symptoms and the difficulty in recognizing amebas in biopsied tissues, most cases are not diagnosed or effectively treated, leading to a >95% mortality. We report here on five cases of balamuthiasis that were diagnosed by indirect immunofluorescence (IIF) staining of serum for anti-
Balamuthia
antibodies (titer ≥ 1:128) and confirmed by IIF of unstained brain tissue sections and/or detection of amebas in hematoxylin-eosin-stained slides. Additionally, we have used the PCR for the detection of mitochondrial 16S rRNA gene DNA from the ameba in clinical specimens such as brain tissue and cerebrospinal fluid (CSF) from individuals with
Balamuthia
encephalitis.
Balamuthia
DNA was successfully detected by the PCR in clinical samples from all five individuals. It was detected in brain tissue from three cases, in CSF from three cases, and in one of two samples of lung tissue from two individuals, but not in two samples of kidney tissue tested. One sample of unfixed brain tissue was culture positive for
Balamuthia
. In order to test the sensitivity of the PCR for detection of
Balamuthia
DNA, CSF specimens from two individuals negative for amebic infection were spiked with
Balamuthia
amebas. We found that it was possible to detect
Balamuthia
DNA in the PCR mixtures containing mitochondrial DNA from 1 to as little as 0.2 ameba per reaction mixture. A single
Balamuthia
ameba contains multiple mitochondrial targets; thus, 0.2 ameba represents multiple targets for amplification and is not equivalent to 0.2 of an ameba as a target.
Frequent Carriage of Panton-Valentine Leucocidin Genes by
Staphylococcus aureus
Isolates from Surgically Drained Abscesses
Authors: Issartel Bertrand , Tristan Anne , Lechevallier Sylvain , Bruyère Franck , Lina Gerard , Garin Benoît , Lacassin Flore , Bes Michèle , Vandenesch François , Etienne Jerome
Published: July 2005

Between 1 February and 15 April 2002, 95 patients were admitted to Gaston Bourret Territorial Hospital (New Caledonia, France) for drainage of community-acquired soft tissue abscesses.
Staphylococcus aureus
was detected in 68 cases (72%). Two-thirds of the patients with
S. aureus
infection had furuncles, which were located on the limbs in 82% of cases. The median interval between symptom onset and hospital admission was 5.7 days. Three-quarters of the patients were Melanesians living in tribes. Fifty-four
S. aureus
isolates were screened for toxin genes. Panton-Valentine leucocidin (PVL) genes were detected in 48 isolates (89%), the exfoliative toxin A gene was detected in 1 isolate, and no toxin genes were detected in 4 isolates.
S. aureus
nasal carriage was detected in 39.7% of patients with
S. aureus
infections. Two infecting
S. aureus
strains and two nasal carriage strains were resistant to methicillin. Comparative pulsed-field gel electrophoresis, performed in 16 cases, showed that five of six patients with PVL-positive nasal carriage strains were infected by the same strains. In contrast, 8 of 10 patients with PVL-negative nasal carriage strains were infected by PVL-positive strains. PVL genes thus appear to be a major virulence factor in both primary and secondary
S. aureus
skin infections.
Quantitative PCR Assay Using Sputum Samples for Rapid Diagnosis of Pneumococcal Pneumonia in Adult Emergency Department Patients
Authors: Yang Samuel , Lin Shin , Khalil Ambreen , Gaydos Charlotte , Nuemberger Eric , Juan George , Hardick Justin , Bartlett John G. , Auwaerter Paul G. , Rothman Richard E.
Published: July 2005

Accurate diagnosis of pneumococcal pneumonia in the acute-care setting remains a challenge due to the inadequate sensitivity of conventional diagnostic tests. Sputum cultures, which are likely to have the highest diagnostic yields of all specimen types, have been considered unreliable, due to their inability to differentiate colonization from infection. Our objective was to evaluate the potential clinical utility of a rapid quantitative real-time PCR assay using sputum samples for
Streptococcus pneumoniae
in adult patients with community-acquired pneumonia (CAP). A prospective clinical observational study of consecutively enrolled emergency department patients with CAP was performed; only those patients with excess good-quality sputum samples were included for evaluation. Sputum samples were tested for the presence of
S. pneumoniae
by using a quantitative PCR that targets the pneumolysin gene. PCR findings were compared with those of a composite reference standard comprising Gram staining of sputum samples and sputum/blood cultures. The area under the curve (AUC) and a log-transformed threshold, which provides the maximal sensitivity and specificity, were calculated. Of 487 subjects enrolled, 129 were evaluable. Receiver operating characteristic curve analysis demonstrated an AUC of 0.87. Sensitivity and specificity were 90.0 percent and 80.0 percent, respectively; positive and negative predictive values were 58.7 percent and 96.2 percent, respectively. We have demonstrated that a quantitative rapid pneumolysin PCR assay has favorable accuracy for diagnosis of pneumococcal pneumonia in adult patients with CAP; this assay may be a useful diagnostic adjunct for clinicians, particularly those practicing in the acute-care setting, where rapid pathogen identification may assist in selection of the most appropriate antibiotics.