Simple Algorithm for Identification of Bordetella pertussis Pertactin Gene Variants

Studies performed in several countries have demonstrated the recent emergence and subsequent dominance of circulating Bordetella pertussis strains harboring pertactin and pertussis toxin variants not included in pertussis vaccines. Determination of the pertactin gene variants is commonly performed using a time-consuming and expensive sequence analysis. We developed a simple and reliable pertactin typing algorithm suitable for large-scale screening. The assay correctly identified all pertactin alleles in representative strains. The typing of 231 clinical strains of B. pertussis routinely isolated in Belgium showed that this algorithm was adequate to identify less-frequent prn types like prn9 and prn11.     

Evaluation of different cytomegalovirus (CMV) DNA PCR protocols for analysis of dried blood spots from consecutive cases of neonates with congenital CMV infections

Two protocols for the extraction of cytomegalovirus (CMV) DNA and two methods for the amplification of CMV DNA in dried blood spots were evaluated for the retrospective diagnosis of congenital CMV infection. During the period from 1996 to 2006, a urine screening program detected 76 congenitally infected neonates. Stored Guthrie cards with blood from 55 cases and 12 controls were tested. Two spots of dried blood were cut from each card and evaluated in two centers. CMV DNA was extracted from a whole single spot. Center 1 used phenol-chloroform extraction and ethanol precipitation followed by a conventional PCR. Center 2 used the NucliSens easyMAG automated DNA/RNA extraction platform (bioMérieux) followed by a real-time PCR. For evaluation of the extraction method, DNA extracted from each blood spot was evaluated by the amplification method used by the collaborating center. The sensitivities were 66% for center 1 and 73% for center 2. None of the controls were positive. A sensitivity as high as 82% could be obtained by combining the most sensitive extraction method (the phenol-chloroform procedure) with the most sensitive PCR method (real-time PCR). The detection rate was not influenced by the duration of storage of the spots. The sensitivity was higher with blood from congenitally infected cases due to a primary maternal CMV infection, regardless of the protocol used. However, the difference reached significance only for the least-sensitive protocol (P = 0.036).     

Use of anticancer agents in gynecological oncology during pregnancy: a systematic review of maternal pharmacokinetics and transplacental transfer

Introduction: Cancer affects one in a thousand pregnant women and gynecological cancers are one of the most frequent malignancies. Chemotherapy remains the cornerstone treatment for gynecological cancer. Although all chemotherapeutic agents can cross the placental barrier, the extent of placental transfer varies considerably. Furthermore, the significant physiological variations observed in pregnant women may have an impact on pharmacokinetic parameters. Given the complexity of predicting placental transfer, in vivo and ex vivo studies are essential in this context. In view of the paucity of data on chemotherapy during pregnancy, the objective of the present study was to summarize all the available data on the transplacental transfer of anticancer drugs used to treat gynecological cancers.

Areas covered: In order to evaluate the in vivo and ex vivo transplacental transfer of the anticancer drugs most frequently used in gynecological malignancies, we carried out a comprehensive review of the literature published from 1967 to 2015. Lastly, we summarized recent clinical guidelines on the treatment of gynecological cancers in pregnant patients.

Expert opinion: The preclinical and scarce clinical data must now be extrapolated to define the maternofetal toxicity/efficacy profile and thus guide the physicians to choose anticancer drugs more efficiently in this complex situation.     

Endothelial exosomes contribute to the antitumor response during breast cancer neoadjuvant chemotherapy via microRNA transfer

The interaction between tumor cells and their microenvironment is an essential aspect of tumor development. Therefore, understanding how this microenvironment communicates with tumor cells is crucial for the development of new anti-cancer therapies. MicroRNAs (miRNAs) are small non-coding RNAs that inhibit gene expression. They are secreted into the extracellular medium in vesicles called exosomes, which allow communication between cells via the transfer of their cargo. Consequently, we hypothesized that circulating endothelial miRNAs could be transferred to tumor cells and modify their phenotype. Using exogenous miRNA, we demonstrated that endothelial cells can transfer miRNA to tumor cells via exosomes. Using miRNA profiling, we identified miR-503, which exhibited downregulated levels in exosomes released from endothelial cells cultured under tumoral conditions. The modulation of miR-503 in breast cancer cells altered their proliferative and invasive capacities. We then identified two targets of miR-503, CCND2 and CCND3. Moreover, we measured increased plasmatic miR-503 in breast cancer patients after neoadjuvant chemotherapy, which could be partly due to increased miRNA secretion by endothelial cells. Taken together, our data are the first to reveal the involvement of the endothelium in the modulation of tumor development via the secretion of circulating miR-503 in response to chemotherapy treatment.     

Association of sleep and anaesthesia EEG biomarkers with preoperative MoCA score: A pilot study

Introduction

Preoperative cognitive impairments increase the risk of postoperative complications. The electroencephalogram (EEG) could provide information on cognitive vulnerability. The feasibility and clinical relevance of sleep EEG (EEG_sleep) compared to intraoperative EEG (EEG_intraop) in cognitive risk stratification remains to be explored. We investigated similarities between EEG_sleep and EEG_intraop vis-a-vis preoperative cognitive impairments.

Methods

Pilot study including 27 patients (63 year old [53.5, 70.0]) to whom Montreal cognitive assessment (MoCA) and EEG_sleep were administered 1 day before a propofol-based general anaesthesia, in addition to EEG_intraop acquisition from depth-of-anaesthesia monitors. Sleep spindles on EEG_sleep and intraoperative alpha-band power on EEG_intraop were particularly explored.

Results

In total, 11 (41%) patients had a MoCA <25 points. These patients had a significantly lower sleep spindle power on EEG_sleep (25 vs. 40 μv²/Hz, p = .035) and had a weaker intraoperative alpha-band power on EEG_intraop (85 vs. 150 μv²/Hz, p = .001) compared to patients with normal MoCA. Correlation between sleep spindle and intraoperative alpha-band power was positive and significant (r = 0.544, p = .003).

Conclusion

Preoperative cognitive impairment appears to be detectable by both EEG_sleep and EEG_intraop. Preoperative sleep EEG to assess perioperative cognitive risk is feasible but more data are needed to demonstrate its benefit compared to intraoperative EEG.     

Control of B cell lymphoma recognition via natural killer inhibitory receptors implies a role for human Vγ9/Vδ2 T cells in tumor immunity

The Vγ9/Vδ2 T cell receptor (TCR) is expressed by most human γδ T cells. We show here that cytotoxic T lymphocytes of the Vγ9/Vδ2 subset, but not of the Vδ1 subset of human γδ T cells, express natural killer inhibitory receptors (KIR) with specificity for different HLA class I alleles that down-regulate TCR-mediated signaling in response to HLA class I-expressing B cell lymphomas. Vγ9/Vδ2 T cell clones with a T helper cell phenotype lack KIR and produce lymphokines in response to most human B cell lymphomas, just as they do upon recognition of the HLA class l-deficient human Burkitt's lymphoma Daudi. Thus, human Vγ9/Vδ2 T cells have an innate specificity for nonpolymorphic cell surface structures expressed by many lymphoma cells and their cytotoxic activity is controlled by KIR. These results imply a general role of human Vγ9/Vδ2 T cells in the defense against hematopoietic tumors that is distinct from NK cells.     

Ten-year outcomes of islet transplantation in patients with type 1 diabetes: Data from the Swiss-French GRAGIL network

To describe the 10-year outcomes of islet transplantation within the Swiss-French GRAGIL Network, in patients with type 1 diabetes experiencing high glucose variability associated with severe hypoglycemia and/or with functional kidney graft. We conducted a retrospective analysis of all subjects transplanted in the GRAGIL-1c and GARGIL-2 islet transplantation trials and analyzed components of metabolic control, graft function and safety outcomes over the 10-year period of follow-up. Forty-four patients were included between September 2003 and April 2010. Thirty-one patients completed a 10-year follow-up. Ten years after islet transplantation, median HbA1c was 7.2% (6.2–8.0) (55 mmol/mol [44–64]) versus 8.0% (7.1–9.1) (64 mmol/mol [54–76]) before transplantation (p < .001). Seventeen of 23 (73.9%) recipients were free of severe hypoglycemia, 1/21 patients (4.8%) was insulin-independent and median C-peptide was 0.6 ng/ml (0.2–1.2). Insulin requirements (UI/kg/day) were 0.3 (0.1–0.5) versus 0.5 (0.4–0.6) before transplantation (p < .001). Median (IQR) β-score was 1 (0–4) (p < .05 when comparing with pre-transplantation values) and 51.9% recipients had a functional islet graft at 10 years. With a 10-year follow-up in a multicentric network, islet transplantation provided sustained improvement of glycemic control and was efficient to prevent severe hypoglycemia in almost 75% of the recipients.     

Comparison of 3D multi-echo gradient-echo and 2D T2* MR sequences for the detection of arterial thrombus in patients with acute stroke

Objectives

We compared a multi-echo gradient-echo magnetic resonance sequence (susceptibility-weighted angiography [SWAN]) with the T2* sequence for the detection of an arterial thrombus in acute ischaemic stroke.

Methods

Seventy-four consecutive patients with acute ischaemic stroke were included. Proximal arterial occlusions were diagnosed using time-of-flight (TOF) magnetic resonance angiography (MRA). Two-dimensional (2D) axial reformats from 3D SWAN were generated to match with 2D T2* images. For arterial thrombus detection, each set of MR images (T2*, 2D SWAN reformats and 3D multiplanar SWAN images) was examined independently and separately by three observers who assigned the images to one of three categories: (0) absence of thrombus, (1) uncertain thrombus, (2) certain thrombus. Agreement and diagnostic accuracy were calculated.

Results

Twenty-four proximal arterial occlusions involving the anterior (n?=?20) or posterior (n?=?4) circulation were found. Inter-observer agreement was moderate using T2* images (κ?=?0.58), good using 2D SWAN reformats (κ?=?0.83) and excellent using multiplanar SWAN images (κ?=?0.90). For the diagnosis of thrombus, T2* images were 54 % sensitive and 86 % specific, 2D SWAN reformats were 83 % sensitive and 94 % specific and SWAN multiplanar analysis was 96 % sensitive and 100 % specific.

Conclusions

Three-dimensional SWAN sequence improves the detection of arterial thrombus in patients with acute ischaemic stroke in comparison with the 2D T2* sequence.

Key Points

? Multi-echo gradient-echo MR (e.g. susceptibility-weighted angiograph, [SWAN]) is increasingly used in neuroradiology. ? Compared with conventional T2* sequences, SWAN improves detection of arterial thrombus. ? Multiplanar SWAN analysis had the best diagnostic performance for arterial thrombus detection. ? Sensitivity was 96?% and specificity 100?%. ? Findings support combination of time-of-flight and susceptibility effects in suspected acute stroke.