Sensitivity and specificity of the VIKIA Rota-Adeno immuno-chromatographic test (bioMérieux) and the ELISA IDEIA Rotavirus kit (Dako) compared to genotyping

The performances of two diagnostic tests for rotavirus infection in stool samples were evaluated during a prospective study in children of less than 36 months in child-care centers of Lyon from November 2004 to May 2005. The VIKIA Rota-Adeno immuno-chromatographic test (bioMérieux) and the ELISA IDEIA Rotavirus kit (Dako) were compared with a referral method, the genotyping. Fifty-seven stool samples were collected and analyzed by RT-PCR. The virus genome was detected in 29 samples. The most frequent genotypic combinations were G9P[8] with a prevalence of 75.9%. Sensitivity and specificity of the VIKIA Rota-Adeno test and the ELISA IDEIA Rotavirus kit were strictly comparable and very good: 96.6% (83.0; 99.9) and 96.4% (81.6; 99.9), respectively. The immuno-chromatographic technique were in concordance with the ELISA tests in 93.6% of cases. Thus, the VIKIA Rota-Adeno test is a good alternative for the occasional analysis of stool samples in ambulatory practice.     

Evaluation of real-time RT-PCR Rhino&EV/Cc r-gene® (bioMérieux) kit versions 1 and 2 for rhinovirus detection

BackgroundHuman rhinoviruses (HRVs) are frequent etiologic agents of tract infections, ranging from benign upper to potentially life-threatening lower respiratory tract infections. Diagnosis is based on molecular methods. 169 HRV types, belonging to species A, B and C, have been identified. This high genetic diversity makes it difficult to accurately detect circulating HRVs and to diagnose severe infection.ObjectivesTo comparatively assess the ability to detect HRV clinical isolates of the first version (V1) of the commercial real-time RT-PCR Rhino&EV/Cc r-gene^® (bioMérieux) kit, of an in-house RT-PCR followed by genotyping, considered as the reference method, and of the second version of this commercial test (V2).Study designFrom September 2011 to April 2013, HRVs were prospectively detected in 2525 respiratory specimens, using V1 in combination with the in-house reference RT-PCR. In November 2013, 85 specimens that had given initially false negative results with V1 were retested simultaneously with V1 and V2 and the in-house RT-PCR. In addition, 421 negative specimens with the in-house assay were prospectively tested with V2.ResultsAmong the 2525 specimens, V1 detected 80.7% (502/622) of in-house RT-PCR positive isolates: 85.3% (220/258) of HRV-A, 84.4% (27/32) of HRV-B and 74.9% (176/235) of HRV-C. Among the 85 respiratory samples tested with V1, V2 and the in-house RT-PCR, V2 was more efficient than V1 in detecting 16 HRV isolates: 11/33 (33.3%) of HRV-A and 5/47 (10.6%) of HRV-C tested. The analytical sensitivity of V2 was greater for 8/18 HRV-A genotypes and 2/22 HRV-C genotypes. Relative to the in-house assay, the specificity of V2 was 100% (421/421).ConclusionsThis study showed a slightly higher sensitivity of V2. However, diverse genotypes, especially HRV-C, were undetected.     

Conservative treatment in epithelial ovarian cancer: results of a multicentre study of the GCCLCC (Groupe des Chirurgiens de Centre de Lutte Contre le Cancer) and SFOG (Société Francaise d'Oncologie Gynécologique)

BACKGROUND: Results of conservative management of epithelial ovarian cancer (EOC) remain controversial in the literature. The aim of this study was to assess the clinical outcomes and fertility following fertility-sparing surgical management of EOC in a retrospective multicentre study. METHODS: A multicentre retrospective study was performed by members of two French groups. Six inclusion criteria were defined: (i) Histological review by the same pathologist; (ii) age < or =40 years; (iii) conservative management; (iv) complete peritoneal staging; (v) delivery of a platinum-based chemotherapy in stage > or = IC; and (vi) follow-up >1 year. RESULTS: Thirty-four patients fulfilled the inclusion criteria: 30 had stage IA disease; three had stage IC and one had stage IIA. Eleven patients had recurrence: 10 patients had invasive disease and one had borderline recurrence. Among 10 patients with invasive recurrence, initial stage and grade were: stage IA G1, n = 1; stage IA G2, n = 4; stage IA G3, n = 1; and stage> or = IC, n = 4. All patients with stage > IA had recurrence. Ten pregnancies were observed in nine patients. CONCLUSION: Conservative surgery for patients with EOC could be considered in young patients with stage IA G1 disease. This procedure should not be performed in patients with FIGO stage > IA.     

Effect of aging on spontaneous micronucleus frequencies in peripheral blood of nine mouse strains: the results of the 7th collaborative study organized by CSGMT/JEMS · MMS: Mutation Res. 338 (1995) 51–57

The spontaneous frequencies of micronucleated reticulocytes (MNRETs) were examined monthly over the life spans of animals beloning to nine mouse strains for the 7th collaborative study organized by the CSGMT/JEMS · MMS. Both sexes of the BDF1 strain and females of the A/J strain showed a statistically significant increase ni mean spontaneous MNRET frequency in their last month of life, suggesting the possibility of strain-specific, age-dependent chromosomal instability. SAMP6/Tan, an accelerated senescence-prone strain, showed the same tendency, although it was not statistically significant. The other strains studied, ddY, CD-1, B6C3F1, SAMR1, and MS/Ae, did not show significant age-related differences in mean of MNRET frequencies. More extensive statistical analyses are underway, and the outcomes will be reported separately.     

ITS–RFLP- and RAPD-based genetic variability of Trypanosoma cruzi I, human and vector strains in Santander (Colombia)

Chagas disease is a severe public health problem in Latin-American countries. In Colombia, the predominance of Trypanosoma cruzi I has been described in the literature, with a broad heterogeneity between strains. However, most of the studies carried out centered on isoenzyme analysis, with a smaller number that focus on other molecular methods. In this report, we discuss the results of a molecular analysis of T. cruzi I strains, isolated from the domestic cycle, from the department of Santander, one of the territorial divisions where the prevalence of infection is highest. Internal transcribed spacer–restriction fragment length polymorphism and random amplification of polymorphic DNA were used to characterize 16 strains from human and vector (Triatominae) hosts. The data reveal a clustering based on the biological origin. Human and vector strains grouped separately; however, three vector strains clustered together with human strains. These results indicate that genetic differences exist in the strains that infect both hosts. We conclude that T. cruzi I populations in the domestic cycle of transmission of Chagas disease in Santander are heterogeneous and are composed of different clones. The epidemiological and biological implications are discussed.     

Evaluation of the methodological quality of the Rémic (microbiology guidelines - bacteriology and mycology) of the Société française de microbiologie

We have evaluated the methodological quality of the Rémic (microbiology guidelines?-?bacteriology and mycology) of the Société fran?aise de microbiologie (edition2007), using to AGREE criteria, which are consensual at an international level, in particular at the the World Health Organisation (WHO) and at the European Union. The methodological quality of the Rémic appears to be sub-optimal. These shortcomings in quality are mainly observed in AGREE domain n°?5 (applicability), in AGREE item n°?5 (patients' opinions were not considered), and in AGREE item n°?23 (conflicts of interest were not declared). The users of the Rémic must be aware of these few methodological shortcomings in order for them to be careful before they put its recommendation in practice. In conclusion, we advise the editors of the Rémic to insert at least a methodological chapter in their next edition.     

Envenomation accidents caused by snakebites in the surgical emergency unit of Gabriel-Touré Hospital, Bamako, Mali (1998-1999)

This study aimed at assessing envenomations caused by snakebite in the Gabriel Touré hospital between January 1998 and December 1999. We included in total 112 victims of snakebites proved by the identification of the snake, the fangs traces and envenomation signs. The bites concerned 34 women (30.36%) versus 78 men (69.64%) from the Bamako district, while 66.7% were from a rural environment. The farmers were the most affected category (35.1%). The bite concerned the lower limb in 83.3% of the cases. Snakes such as Echis ocellatus, Bitis arietans, Naja nigricollis and N. katiensis caused the envenomations. The viperine syndrome dominated the clinical picture; its severity depended on the bleeding stage and the delay of administration of the antivenom serum (AVS) after the envenomation: 19 hours for Echis sp. and 2 hours for Naja sp. This study shows that the envenomation accidents caused by snakes are frequent with a lethality rate of 9.8%.     

Interleukin-6 and the IL-6 (–174) C/G polymorphism in breast pathologies and in HIV-infected patients

Introduction

Breast cancer and acquired immunodeficiency syndrome (AIDS) are key issues for modern medicine. The aim of the current study was to present how cytokines, in the example of IL-6 and its polymorphism, can affect these two conditions.

Material and methods

Thirty-one women with benign breast tumours, 42 breast cancer patients and 40 HIV-infected females were enrolled in the study. Serum IL-6 levels were determined by ELISA. The IL-6 polymorphism was genotyped by PCR-RFLP.

Results

Serum IL-6 in patients with benign breast tumours was significantly lower than in females with breast cancer (p = 0.017) and HIV-infected women (p = 0.032). We did not find statistically significant differences in serum IL-6 level between females with breast cancer and HIV-infected women (p = 0.749). Comparing the distribution of genotypes and frequency of the IL-6 (–174) C/G polymorphism between the three study groups – breast cancer patients, patients with benign breast tumours, and HIV-infected patients – we did not find any statistically significant differences.

Conclusions

IL-6 can play an important role in pathogenesis of breast cancer and HIV infection and its level is higher than in the control group irrespective of distribution of genotypes and frequency of the IL-6 (–174) C/G polymorphism.     

Activation of PI 3-kinase/Akt/NF-κB and Stat3 signaling by avian reovirus S1133 in the early stages of infection results in an inflammatory response and delayed apoptosis

Avian reovirus (ARV) strain S1133 causes apoptosis in host cells in the middle to late stages of infection. This study investigated the early-stage biological response and intracellular signaling in ARV S1133-infected Vero and chicken cells. Treatment with conditioned medium from ARV S1133-infected cells increased the chemotactic activity of U937 cells. Neutralizing antibodies against IL-1β and IL-6 showed that both cytokines contribute to viral-induced inflammation but neither affect cell survival. Inhibition of Akt, NF-κB, and Stat3 released the chemotactic activity and anti-apoptotic effect elicited by ARV S1133. ARV S1133 activated PI 3-kinase-dependent Akt/NF-κB and p70 S6 kinase, as well as Stat3; however, p70 S6 kinase was not involved in ARV S1133-mediated effects. DF1 cells over-expressing constitutively active PI 3-kinase and Stat3 showed association with enhancement of anti-apoptotic activity. In conclusion, in the early stages of ARV S1133 infection, activation of cell survival signals contributes to virus-induced inflammation and anti-apoptotic response.     

Subunits α, β and γ of the epithelial Na+ channel (ENaC) are functionally related to the hypertonicity-induced cation channel (HICC) in rat hepatocytes

Specific small interfering RNA (siRNA) constructs were used to test for the functional relation of subunits α, β, and γ of the epithelial Na⁺ channel (ENaC) to the hypertonicity-induced cation channel (HICC) in confluent rat hepatocytes. In current-clamp recordings, hypertonic stress (300 → 400 mosM) increased membrane conductance from 75.4 ± 9.4 to 91.1 ± 11.2 pS (p < 0.001). The effect was completely blocked by 100 μM amiloride and reduced to 46, 30, and 45% of the control value by anti-α-, anti-β-, and anti-γ-rENaC siRNA, respectively. Scanning acoustic microscopy revealed an initial shrinkage of cells from 6.98 ± 0.45 to 6.03 ± 0.43 pl within 2 min. This passive response was then followed by a regulatory volume increase (RVI) by 0.42 ± 0.05 pl (p < 0.001). With anti-α-, anti-β-, and anti-γ-rENaC siRNA, the volume response was reduced to 31, 31, and 36% of the reference level, respectively. It is concluded that all three subunits of the ENaC are functionally related to RVI and HICC activation in rat hepatocytes.     

Working memory deficits and related disinhibition of the cAMP/PKA/CREB are alleviated by prefrontal α4β2*-nAChRs stimulation in aged mice

The present study investigates in aged mice the working memory (WM) enhancing potential of the selective α4β2* nicotinic receptor agonist S 38232 as compared with the cholinesterase inhibitor donepezil, and their effect on cAMP response element binding protein (CREB) phosphorylation (pCREB) as a marker of neuronal activity. We first showed that aged mice exhibit a WM deficit and an increase of pCREB in the prelimbic cortex (PL) as compared with young mice, whereas no modification appears in the CA1. Further, we showed that systemic administration of S 38232 restored WM in aged mice and alleviated PL CREB overphosphorylation. Donepezil alleviated age-related memory deficits, however, by increasing pCREB in the CA1, while pCREB in PL remained unaffected. Finally, whereas neuronal inhibition by lidocaine infusion in the PL appeared deleterious in young mice, the infusion of Rp-cAMPS (a compound known to inhibit CREB phosphorylation) or S 38232 rescued WM in aged animals. Thus, by targeting the α4β2*-nicotinic receptor of the PL, S 38232 alleviates PL CREB overphosphorylation and restores WM in aged mice, which opens new pharmacologic perspectives of therapeutic strategy.     

Deletion of the D domain of the human parainfluenza virus type 3 (HPIV3) PD protein results in decreased viral RNA synthesis and beta interferon (IFN-β) expression

Parvoviridae is a family of small non-enveloped viruses and divided into two subfamilies. The family members infect a wide range of organisms from insects to humans and some of the members (e.g., nonpathogenic adeno-associated viruses) are effective gene therapy delivery vectors. We detailed the synonymous codon usage pattern of Parvoviridae family from the available 58 sequenced genomes through multivariate statistical methods. Our results revealed that nine viruses showed some degree of strong codon bias, and the others possessed a general weak trend of codon bias. ENc-plot and neutrality plot results showed that selective pressure dominated over mutation in shapes coding sequence’s composition. The overall GC content and GC content at the third synonymous codon position were the principal determinants behind the variations within the codon usage patterns, as they both significantly correlated with the first axis of correspondence analysis. In addition, gene length had no direct influence on the codon usage pattern. Densovirinae subfamily and Parvovirinae subfamily possessed nine identical preferred codons, though most of the two subfamilies codon usage frequencies were significantly different. The result of cluster analysis based on synonymous codon usage was discordant with that of taxonomic classification. Adeno-associated viruses formed a separated clade far from other Parvoviridae members in the dendrogram. Thus, we concluded that natural selection rather than mutation pressure accounts for the main factor that affects the codon bias in Parvoviridae family.     

Influenza virus infections in patients with malignancies –– characteristics and outcome of the season 2014/15. A survey conducted by the Infectious Diseases Working Party (AGIHO) of the German Society of Haematology and Medical Oncology (DGHO)

Influenza virus infections (IVI) may pose a vital threat to immunocompromised patients such as those suffering from malignancies, but specific data on epidemiology and outcome in these patients are scarce. In this study, we collected data on patients with active cancer or with a history of cancer, presenting with documented IVI in eight centres in Germany. Two hundred and three patients were identified, suffering from haematological malignancies or solid tumours; 109 (54 %) patients had active malignant disease. Influenza A was detected in 155 (77 %) and Influenza B in 46 (23 %) of patients (genera not determined in two patients). Clinical symptoms were consistent with upper respiratory tract infection in 55/203 (27 %), influenza-like illness in 82/203 (40 %), and pneumonia in 67/203 (33 %). Anti-viral treatment with oseltamivir was received by 116/195 (59 %). Superinfections occurred in 37/203 (18 %), and admission on an intensive care unit was required in 26/203 (13 %). Seventeen patients (9 %) died. Independent risk factors for death were delayed diagnosis of IVI and bacterial or fungal superinfection, but not underlying malignancy or ongoing immunosuppression. In conclusion, patients with IVI show high rates of pneumonia and mortality. Early and rapid diagnosis is essential. The high rate of pneumonia and superinfections should be taken into account when managing IVI in these patients.     

Performance of Two Resin-Containing Blood Culture Media in Detection of Bloodstream Infections and in Direct Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS) Broth Assays for Isolate Identification: Clinical Comparison of the BacT/Alert Plus and Bactec Plus Systems

We compared the clinical performances of the BacT/Alert Plus (bioMérieux) and Bactec Plus (Becton Dickinson) aerobic and anaerobic blood culture (BC) media with adsorbent polymeric beads. Patients ≥16 years old with suspected bloodstream infections (BSIs) were enrolled in intensive care units and infectious disease wards. A single 40-ml blood sample was collected from each and used to inoculate (10 ml/bottle) one set of BacT/Alert Plus cultures and one set of Bactec Plus cultures, each set consisting of one aerobic and one anaerobic bottle. Cultures were incubated ≤5 days in the BacT/Alert 3D and Bactec FX instruments, respectively. A total of 128 unique BSI episodes were identified based on the recovery of clinically significant growth in 212 aerobic cultures (106 BacT/Alert and 106 Bactec) and 151 anaerobic cultures (82 BacT/Alert and 69 Bactec). The BacT/Alert aerobic medium had higher recovery rates for Gram-positive cocci (P = 0.024), whereas the Bactec aerobic medium was superior for recovery of Gram-negative bacilli (P = 0.006). BacT/Alert anaerobic medium recovery rates exceeded those of the Bactec anaerobic medium for total organisms (P = 0.003), Gram-positive cocci (P = 0.013), and Escherichia coli (P = 0.030). In terms of capacity for diagnosing the 128 septic episodes, the BacT/Alert and Bactec sets were comparable, although the former sets diagnosed more BSIs caused by Gram-positive cocci (P = 0.008). They also allowed earlier identification of coagulase-negative staphylococcal growth (mean, 2.8 h; P = 0.003) and growth in samples from patients not on antimicrobial therapy that yielded positive results (mean, 1.3 h; P < 0.001). Similarly high percentages of microorganisms in BacT/Alert and Bactec cultures (93.8% and 93.3%, respectively) were identified by direct matrix-assisted laser desorption ionization–time of flight mass spectrometry assay of BC broths. The BacT/Alert Plus media line appears to be a reliable, timesaving tool for routine detection of BSIs in the population we studied, although further studies are needed to evaluate their performance in other settings.Throughout the world, the number of patients at risk for bloodstream infections (BSIs) continues to rise (1). BSIs are associated with high rates of morbidity and mortality, and they markedly increase the costs of hospital care (1, 2). Prompt identification of the causative agent(s) and rapid initiation of appropriate antimicrobial therapy are critical for reducing mortality, especially in patients with septic shock (2,–4). Blood culture (BC) remains the gold standard for diagnosing BSIs (5, 6). Over the decades, improvements in culture media and the availability of software-assisted, automated growth detectors have enhanced the recovery of bloodstream pathogens and decreased the time to detection (TTD) of microbial growth. The time-consuming process of isolate identification with conventional culture-based methods has also been improved, thanks to the development of new methods that can be used directly on broth samples from BC bottles, such as matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry (MS) (7). Various studies have shown this approach to be a reliable, cost-effective, timesaving alternative for routine identification of bacteria and yeasts causing BSIs (7,–10). Its routine use can increase the proportion of patients who receive effective antimicrobial treatment within 24 h of BC positivity, and its potential impact on outcome is even greater than that of Gram-stain reporting (4, 11).Despite these advances, however, limitations persist. One of the most important involves the diagnostic performance of BCs collected from patients who are already on antimicrobial therapy. In up to 87% of patients with severe sepsis, empirical antimicrobial therapy is started before blood samples for cultures are drawn, and this practice can reduce or delay pathogen recovery (12,–15). Increasing the diagnostic yield of positive BCs in this setting would allow more effective management of a highly vulnerable patient population (12, 16).To this end, several manufacturers have designed BC media containing resins (Bactec Plus Aerobic/F and Anaerobic/F bottles; Becton Dickinson Instrument Systems, Sparks, MD) or charcoal (BacT/Alert FAN, FA, and FN bottles; bioMérieux, Marcy l''Etoile, France) designed to adsorb antimicrobial drugs present in the blood. The ability of these media to improve the detection of bacteremia and fungemia in patients on antimicrobial therapy has been widely documented (13, 14, 17,–28). In earlier studies, resin- and charcoal-based systems displayed similar recovery rates and TTDs, regardless of whether antibiotics were being administered when the BCs were drawn (13, 27, 28), but two more recent studies suggested that, in the presence of antibiotics, resin-containing media are superior (14, 15). In several studies, both types of media improved recovery and reduced the TTDs of significant pathogens in simulated BCs inoculated with blood containing therapeutic levels of commonly used antibiotics and antifungal agents (29,–37). However, significant differences emerged between the resin- and charcoal-based media relative to specific microorganism-antimicrobial combinations. Charcoal particles also appear to interfere with the reading of Gram-stained smears, and charcoal-containing broths must be subjected to more complex, time-consuming extraction protocols before undergoing direct MALDI-TOF MS assay for species-level isolate identification (28, 38, 39).Three recent studies (40,–42) found that the new bioMérieux BacT/Alert FA Plus, FN Plus, and PF Plus BC media, which contain adsorbent polymeric beads, improve and accelerate the detection of bloodstream pathogens compared with standard BacT/Alert media and with BacT/Alert media containing charcoal particles. The present study represents the first attempt to evaluate the performance of the new BacT/Alert FA Plus and FN Plus media compared with Becton Dickinson''s Bactec Plus Aerobic/F and Anaerobic/F media, which also contain antimicrobial-binding resins.(Portions of the data from this study were presented at the 23rd European Congress of Clinical Microbiology and Infectious Diseases, Berlin, Germany 2013.)