A Multiplex PCR and DNA-Sequencing Workflow on Serum for the Diagnosis and Species Identification for Invasive Aspergillosis and Mucormycosis.

There has been significant increase in the use of molecular tools for the diagnosis of invasive aspergillosis (IA) and mucormycosis. However, their range of detection may be too limited as species diversity and coinfections are increasing. Here, we aimed to evaluate a molecular workflow based on a new multiplex PCR assay detecting the whole Aspergillus genus and the Mucorales order followed by a species-specific PCR or a DNA-sequencing approach for IA and/or mucormycosis diagnosis and species identification on serum. Performances of the MycoGENIE Aspergillus spp./Mucorales spp. duplex PCR kit were analyzed on a broad range of fungal strains and on sera from high-risk patients prospectively over a 12-month period. The kit allowed the detection of nine Aspergillus species and 10 Mucorales (eight genera) strains assessed. No cross-reactions between the two targets were observed. Sera from 744 patients were prospectively analyzed, including 35 IA, 16 mucormycosis, and four coinfections. Sensitivity varies from 85.7% (18/21) in probable/proven IA to 28.6% (4/14) in COVID-19-associated pulmonary aspergillosis. PCR-positive samples corresponded to 21 A. fumigatus, one A. flavus, and one A. nidulans infections. All the disseminated mucormycosis were positive in serum (14/14), including the four Aspergillus coinfections, but sensitivity fell to 33.3% (2/6) in localized forms. DNA sequencing allowed Mucorales identification in serum in 15 patients. Remarkably, the most frequent species identified was Rhizomucor pusillus (eight cases), whereas it is barely found in fungal culture. This molecular workflow is a promising approach to improve IA and mucormycosis diagnosis and epidemiology.

Atom probe tomography quantification of carbon in silicon.

Atom Probe Tomography (APT) was used to quantify carbon in implanted silicon at two various electric fields (~ 15 and 20 V/nm). Using equal proportions of implanted 12C and 13C, the numerous molecular ions that were observed were identified and their contribution to the carbon content statistically derived. Much more accurate carbon quantification was obtained in the lowest electric field analysis by comparing APT with Secondary Ion Mass Spectroscopy profiles. This was assigned to a lower amount of molecular ion dissociations. Furthermore, the number of self-interstitials trapped per carbon atom in clusters was derived. This value of interest for the microelectronics industry regarding dopant diffusion and implantation induced defects was estimated close to one, in agreement with the expected stoichiometry of the SiC phase present in the phase diagram. However, this was obtained only when using low electric field conditions.

[Invasive pulmonary aspergillosis]. / Aspergillose pulmonaire invasive.

INTRODUCTION: Invasive pulmonary aspergillosis (IPA) is an important cause of morbidity and mortality in a wide range of patients. Early recognition and diagnosis have become a major focus in improving the management and outcomes of this life-threatening disease. BACKGROUND: IPA typically occurs during a period of severe and prolonged neutropenia. However, solid organ transplant recipients, patients under immunosuppressive therapy or hospitalized in intensive care units are also at risk. The diagnosis is suspected in the presence of a combination of clinical, biological and CT scan evidence. The microbiological diagnostic strategy should be adapted to the patient's profile. Conventional methods with culture and species identification remain the standard but early diagnosis has been improved by the use of biomarkers such as galactomannan antigen in serum or in bronchoalveolar lavage. OUTLOOK: The epidemiology of IPA should change with the increased use of antifungal prophylactic regimens and the arrival of targeted therapies. Other microbiological tools, such as PCR and other biomarkers, are currently being assessed. CONCLUSIONS: IPA must be considered in a wide range of patients. Its prognosis remains poor despite progress in the microbiological diagnosis and therapeutic management.

Topologically frustrated ionisation in a water-ammonia ice mixture.

Water and ammonia are considered major components of the interiors of the giant icy planets and their satellites, which has motivated their exploration under high P-T conditions. Exotic forms of these pure ices have been revealed at extreme (~megabar) pressures, notably symmetric, ionic, and superionic phases. Here we report on an extensive experimental and computational study of the high-pressure properties of the ammonia monohydrate compound forming from an equimolar mixture of water and ammonia. Our experiments demonstrate that relatively mild pressure conditions (7.4 GPa at 300 K) are sufficient to transform ammonia monohydrate from a prototypical hydrogen-bonded crystal into a form where the standard molecular forms of water and ammonia coexist with their ionic counterparts, hydroxide (OH-) and ammonium [Formula: see text] ions. Using ab initio atomistic simulations, we explain this surprising coexistence of neutral/charged species as resulting from a topological frustration between local homonuclear and long-ranged heteronuclear ionisation mechanisms.

Unsupervised explorative data analysis of normal human leukocytes and BCR/ABL positive leukemic cells mid-infrared spectra.

We proved the ability of Fourier Transform Infrared microspectroscopy (microFTIR) complemented by Principal Component Analysis (PCA) to detect protein phosphorylation/de-phosphorylation in mammalian cells. We analyzed by microFTIR human polymorphonuclear neutrophil (PMNs) leukocytes, mouse-derived parental Ba/F3 cells (Ba/F3#PAR), Ba/F3 cells transfected with p210(BCR/ABL) (Ba/F3#WT) and expressing high levels of protein tyrosine kinase (PTK), and human-derived BCR/ABL positive K562 leukemic cell sub-clones engineered to differently express receptor-type tyrosine-protein phosphatase gamma (PTPRG). Synchrotron radiation (SR) and conventional (globar) IR sources were used to perform microFTIR respectively, on single cells and over several cells within the same sample. Ex vivo time-course experiments were run, inducing maximal protein phosphorylation in PMNs by 100 nM N-formylated tripeptide fMLP. Within the specific IR fingerprint 1800-850 cm(-1) frequency domain, PCA identified two regions with maximal signal variance. These were used to model and test the robustness of PCA in representing the dynamics of protein phosphorylation/de-phosphorylation processes. An IR signal ratio marker reflecting the homeostatic control by protein kinases and phosphatases was identified in normal leukocytes. The models identified by microFTIR and PCA in normal leukocytes also distinguished BCR/ABL positive Ba/F3#WT from BCR/ABL negative Ba/F3#PAR cells as well as K562 cells exposed to functionally active protein tyrosine phosphatase recombinant protein ICD-Tat transduced in cells by HIV-1 Tat technology or cells treated with the PTK inhibitor imatinib mesylate (IMA) from cells exposed to phosphatase inactive (D1028A)ICD-Tat recombinant protein and untreated control cells, respectively. The IR signal marker correctly reflected the degrees of protein phosphorylation associated with abnormal PTK activity in BCR/ABL positive leukemic cells and in general was inversely related to the expression/activity of PTPRG in leukemic sub-clones. In conclusion, we have described a new, reliable and simple spectroscopic method to study the ex vivo protein phosphorylation/de-phosphorylation balance in cell models: it is suitable for biomedical and pharmacological research labs but it also needs further optimization and its evaluation on large cohorts of patients to be proposed in the clinical setting of leukemia.

[Face and neck lift using a superficial musculo-aponeuro­t­ic system (SMAS) plication: Radiological analysis of tissu displacement, a case report].

Objectives: Face and neck lift (FNL) using SMAS plication is a simple and well-known surgical technique. The principle of smas plication from two fixed points allows an important tissue ascension. Our purpose was to objectively evaluate the SMAS plication results, although a common critic may be due to its early deep tissular displacement. Materials and methods: We present the case of a 65 y.o. female patient who received a FNL. Miniature metal pins were placed on the external surface of the SMAS in order to ensure radiological monitoring. A cephalo­metric and photographic monitoring were performed pre­operatively and on day 1, months 2, 6 and 16 postoperatively . We performed a computed numerised analysis of the movement of the markers in an orthogonal axis. Results: All our bearings kept a constant position in the radiographic follow-up , demons­tra­ting the fixity of SMAS kinked 16 months postoperatively. Clinically the correction remained a good appearance with a very slight relaxation of the superficial tissues (skin and subcutaneous fat). Conclusion: In the case of our patient, the tensioning of the deep facial plan (the SMAS) objectively appeared to be efficient over time, with an overall delay of 16 months. Our protocole appeared to be coherent without any specific complication. It permits to objectively differentiate the relaxation of the deep or superfial tissues after a facial lift procedure.

Use of G-CSF to hasten neutrophil recovery after auto-SCT for AML is not associated with increased relapse incidence: a report from the Acute Leukemia Working Party of the EBMT.

Application of G-CSF in AML is controversial as leukemic blasts may express receptors interacting with the cytokine, which may stimulate leukemia growth. We retrospectively analyzed the impact of G-CSF use to accelerate neutrophil recovery after auto-SCT on outcome. Adults with AML in first CR autografted between 1994 and 2010 were included. Nine hundred and seventy two patients were treated with G-CSF after auto-SCT whereas 1121 were not. BM and PB were used as a source of stem cells in 454 (22%) and 1639 (78%) cases, respectively. The incidence of relapse at 5 years in the BM-auto-SCT group was 38% for patients receiving post-transplant G-CSF and 43% for those not treated with G-CSF, P=0.46. In the PB-auto-SCT cohort, respective probabilities were 48% and 49%, P=0.49. No impact of the use of G-CSF could be demonstrated with respect to the probability of leukemia-free survival: in the BM-auto-SCT group, 51% for G-CSF(+) and 48% for G-CSF(-), P=0.73; in PB-auto-SCT group, 42% for G-CSF(+) and 43% for G-CSF(-), P=0.83. Although G-CSF administration significantly shortened the neutropenic phase, no beneficial effect was observed with regard to non-relapse mortality. In patients with AML, the use of G-CSF after auto-SCT is not associated with increased risk of relapse irrespective of the source of stem cells used.

Severe granulomatous reaction associated with hypercalcemia occurring after silicone soft tissue augmentation of the buttocks: a case report.

UNLABELLED: Liquid silicone is a permanent filler. Its use to augment soft tissues for aesthetic purposes was widespread worldwide in the 1960s. Although initially considered to be biologically inert, this substance may cause, after its injection, an inflammatory granulomatous effect of variable severity and, in very rare cases, a severe hypercalcemia, which can be life threatening. The reported case highlights the well-known physiopathology of hypercalcemia, and the various therapeutic options are discussed. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Investigation of lattice dynamical and dielectric properties of MgO under high pressure by means of mid- and far-infrared spectroscopy.

We investigate the lattice dynamical and dielectric properties of MgO single crystals and powders by measurements in the mid- and far-infrared frequency region under high pressures, ranging up to 21.7 GPa. The shift of the restrahlen region is used to determine the pressure dependence of the transverse and longitudinal optical modes. The analysis of the refractive index in the mid- and far-infrared region allowed us to obtain the pressure behavior of the static and electronic dielectric constants. The transverse effective charge slowly decreases under high pressure, reflecting the stability of MgO. As a consequence, the pressure dependence of the static and electronic dielectric constants is mainly determined by the pressure dependence of the polar phonon frequency and Penn gap, resulting in a pronounced decrease of the former and a moderate decrease of the latter.

Differences in the hippocampal frequency of creatine inclusions between the acute and latent phases of pilocarpine model defined using synchrotron radiation-based FTIR microspectroscopy.

Temporal lobe epilepsy (TLE) is the most common type of epilepsy in adults. Of the animal models developed to investigate the pathogenesis of TLE, the one with pilocarpine-induced seizures is most often used. After pilocarpine administration in animals, three distinct periods--acute, latent, and chronic--can be distinguished according to their behavior. The present paper is the continuation of our previous study which has shown an increased occurrence of creatine inclusions in rat hippocampal formations from the acute phase of pilocarpine-induced status epilepticus (SE) and positive correlation between their quantity and the total time of seizure activity within the observation period. In this paper, we tried to verify if anomalies in hippocampal creatine accumulation were the temporary or permanent effect of pilocarpine-evoked seizures. To realize this purpose, male Wistar rats in the latent phase (3 days after pilocarpine administration) were examined. The results obtained for the period when stabilization of animal behavior and EEG occurs were afterwards compared with ones obtained for the acute phase of pilocarpine-induced SE and for naive controls. To investigate the frequency of creatine inclusions within the hippocampal formation as well as in its selected areas (sectors 1-3 of Ammon's horn (CA1-CA3), dentate gyrus (DG), and hilus of DG) and cellular layers (pyramidal, molecular, multiform, and granular cell layers), synchrotron radiation-based Fourier-transform infrared microspectroscopy was used. The applied technique, being a combination of light microscopy and infrared spectroscopy, allowed us to localize microscopic details in the analyzed samples and provided information concerning their chemical composition. Moreover, the use of a synchrotron source of IR radiation allowed us to carry out the research at the diffraction-limited spatial resolution which, because of the typical size of creatine inclusions (from a few to dozens of micrometers), was necessary for our study. The comparison of epileptic animals in the latent phase with controls showed statistically significant increase in the number of creatine inclusions for most of the analyzed hippocampal regions, all examined cellular layers, as well as the whole hippocampal formation. Moreover, for the hilus of the DG and CA3 area, the number of creatine deposits was higher in the latent than in the acute phase after pilocarpine injection. In light of the obtained results, an anomaly in the hippocampal accumulation of creatine is the long-term effect of pilocarpine-evoked seizures, and the intensity of this phenomenon may increase with time passing from the primary injury.

[Treatment of Dupuytren's contracture by collagenase injection]. / Traitement des brides de la maladie de Dupuytren par la collagénase injectable.

Dupuytren's disease is a disorder of the palmar aponeurosis responsible for apparition of pathological collagen cords that will gradually lead to a digital retraction. These cords cause functional disability to the hand with a real handicap for the patient. No curative treatment exists currently in this disease. Injections of collagenase from Clostridium histolyticum cause lysis of the collagen present in the cords and cause an interruption of the palmar cord. It is a new treatment in the management of Dupuytren's disease. The use of collagenase injection in the treatment of palmar form of Dupuytren's disease gives good results in the short and medium terms. It is a simple, minimally invasive treatment, several studies have proven its effectiveness and it approximates percutaneous needle fasciotomy in its easy use and its indications. Patients should be well informed about local reactions and transient pain with the injection of C. histolyticum collagenase. Diffusion of this technique is still limited by: the assessment of its cost compared to that of other methods of treatment and particularly the needle fasciotomy, and the evaluation of results over the long-term.

Incidence and risk factors of EBV reactivation after unrelated cord blood transplantation: a Eurocord and Société Française de Greffe de Moelle-Therapie Cellulaire collaborative study.

EBV viremia and post-transplantation lymphoproliferative disorders (PTLDs) have been associated with high mortality rates after allogeneic hematopoietic SCT (allo-HSCT). Few retrospective studies, without EBV load monitoring postulated that umbilical cord blood transplantation (UCBT) might be associated with high incidence of EBV events. We retrospectively studied 175 UCBT recipients for whom RQ-PCR was used to monitor EBV blood load at least once a week during the first 3 months after UCBT. Median age was 23 years, 74% had leukemia. Conditioning was myeloablative in 54% and reduced intensity conditioning (RIC) was used in 46%. A total of 24 patients presented an EBV reactivation. For 15 patients, the reactivation occurred during the first 100 days (cumulative incidence: 8%) and included 4 EBV-PTLD. Rituximab as preemptive treatment was used in 12 of these 15 patients. In univariate analysis, the increased risk of early EBV reactivation was associated with RIC in combination with antithymocyte globulin (P=0.03) and previous history of auto-HSCT (P=0.01). Multivariate analysis did not find any independent risk factor. EBV reactivation as time-dependent covariate was not statistically associated with survival. Therefore, EBV events were not major complications after UCBT when EBV load is weekly monitored and preemptive treatment started.

Development of a percutaneous penetration predictive model by SR-FTIR.

This work focused on developing a new evaluation criterion of percutaneous penetration, in complement to Log Pow and MW and based on high spatial resolution Fourier transformed infrared (FTIR) microspectroscopy with a synchrotron source (SR-FTIR). Classic Franz cell experiments were run and after 22 h molecule distribution in skin was determined either by HPLC or by SR-FTIR. HPLC data served as reference. HPLC and SR-FTIR results were compared and a new predictive criterion based from SR-FTIR results, named S(index), was determined using a multi-block data analysis technique (ComDim). A predictive cartography of the distribution of molecules in the skin was built and compared to OECD predictive cartography. This new criterion S(index) and the cartography using SR-FTIR/HPLC results provides relevant information for risk analysis regarding prediction of percutaneous penetration and could be used to build a new mathematical model.

[Intra-osseous ganglion cyst of the carpal bones. A review of the literature underlining the importance of systematic computed tomography]. / Kystes synoviaux intraosseux du carpe : intérêt de la tomodensitométrie systématique pour l'évaluation du risque fracturaire.

The intraosseous ganglion cyst (IOGC) is a benign and lytic bone tumor affecting mostly the metaphyseal and epiphyseal regions of long bones. Its location on the short bones, including the carpal bones has been little reported in the literature. Our review of the literature shows consensus about the surgical techniques to use, but there is currently no real consensus about its pathophysiology, and its diagnostic work-up. Complications related to this lesion (mainly the risk of pathologic fracture) are potentially serious, and can cause irreversible damage. They therefore require accurate assessment to guide the choice of medical or surgical treatment, including a CT scan, which - we believe - is essential.

[PEEK customized implant for skull esthetic reconstruction]. / Reconstruction esthétique de la voûte crânienne par implant sur mesure en PEEK.

INTRODUCTION: Craniotomies provide unsightly sequels. Conservative methods for the removed bone, autologous graft, or use of surgical cement do not always allow restoring cranial symmetry. A customized PolyEtherEtherKetone (PEEK) implant may then be used. This implant is made from 3D CT reconstruction. The PEEK material is biocompatible, conformable, strong enough to protect the brain. TECHNICAL NOTE: The authors report a case of aesthetic reconstruction of a large cranial defect using a customized PEEK implant. Balloon scalp expansion was performed before implant placement. DISCUSSION: The aesthetic results are satisfactory, the postoperative short and simple. PEEK is a good alternative to titanium implants for customized implants; it is easier to use and lighter. This customized implant is well suited to correct unsightly sequels of craniotomy.

[Interest and reliability of frozen section biopsy in the treatment of skin tumors]. / Intérêt et fiabilité de l'examen extemporané dans la prise en charge des tumeurs cutanées.

INTRODUCTION: Association of surgical treatment combined to frozen section biopsies appears to be one of the most appropriate therapeutic solution for the treatment of skin carcinomas. We report our experience on 269 tumors assessed with frozen sections. Our goal was to first study the benefit of a one-time surgical approach, then to better target the tumors eligible for this examination and finally to number the difference between frozen section and final histology studies. MATERIALS AND METHODS: This is a retrospective analysis of 269 tumors, operated on an outpatient basis with frozen section followed later by a final histology study. All being performed by the same team of surgeons and pathologist. Only previously biopsied and diagnosed basal cell or squamous cell tumors were included. The recorded data were: location of tumor; histological type; involved margins; number of cuts; differences between frozen section and final histological studies, which conduct to a two-time surgical approach; number of tumors for which frozen section was impossible during surgery and that lead to a two-time surgical approach; type of reconstruction; number of recurrence. RESULTS: The follow up was 48 months (26.6 to 78.1). Histogical analyses were carried on basal cell carcinoma (92%), squamous cell carcinoma (8%) with a topographic distribution mainly in face and neck (72%). Of the 269 tumors excised, 207 representing 77% had a sufficient safety margin, 62 representing 23% had at least one invaded bank that required further surgical resection. We found seven cases of two-time surgical approach. In three cases, it was secondary to medical decision because of a technical difficulty of histologists. In one case, histological diagnosis can't be obtained by frozen section study. In three cases, we found a difference between frozen section and final study. CONCLUSION: The benefit provided by the frozen section takes its value in the treatment of face and neck tumors, whose optimal margins are sometimes difficult to obtain and minimal scar ransom necessary. A one-time surgery was made possible to us thanks to this fast examination. The low rate of second surgery and recurrence allows us to demonstrate the reliability of this technique.