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.

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 .

[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.

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.

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.

[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.

[Study of skin retraction applied to the treatment of skin tumors. Mapping of the human body]. / Étude de la rétraction cutanée appliquée à la prise en charge des tumeurs cutanées. Cartographie du corps humain.

SUBJECT: Skin, the main organ of the human body, is equipped with own biomechanical characteristics, highly variable depending on intra-individual factors (location, weight status, dermatological diseases…) and interindividual (age, sex…). Despite some recent cutometric studies, our review of the literature shows that there is no currently reliable analytical model representing the biomechanical behavior of the skin. Yet, this is a central issue in dermatology surgery, especially in the treatment of skin tumors, for the proper observance of surgical margins. PATIENTS AND METHODS: We studied prospectively on 75 resection specimens (about 71 patient(s)), for the treatment of skin lesions tumor suspicious or known malignant or benign. Room dimensions were measured before and 5 minutes after excision, leading us to calculate a ratio of retraction of the skin surface. This retraction was correlated with age, gender, tumor type, and anatomic location of the site of excision. RESULTS: The power of retraction of the skin varies significantly by region of the body. It is maximum in the upper limb (hand excluded) and in the cervical region. At the cephalic region, skin of the ear and periorbital skin have capacities of important early retraction. Unlike the lower limb (foot excluded), the back skin of the nose and face appear to be a minimum of shrinkage. Age also seems to change on that capacity shrinkage, sex would have no influence. CONCLUSION: Our study confirms the variations in the ability of skin retraction based on a number of factors. In dermato-oncology, that power retraction could cause significant differences between clinical surgical margins and final pathologist margins. We believe it must be taken into account by the couple surgeon-pathologist, especially in the context of invasive and/or recurrent tumors.

[Correction of the deep nasolabial fold using a temporalis superficialis fascia graft]. / Le comblement du sillon nasogénien par greffon de fascia temporalis superficialis.

SUBJECT: The deep nasolabial fold (NLF) due to age is a frequent complaint from patients in plastic surgery. Various surgical and medical management solutions have been described in literature and are often based on different concepts to justify their interest. We present a new surgical treatment of this region using a temporalis superficialis fascia graft in order to fill the NLF. We also propose a review of the literature about the anatomy and the physiology of this major fold of the face. PATIENTS AND METHODS: From 1990 to 2010, we practiced the replenishing of the NLF using a temporalis superficialis fascia graft on 24 female patients. They all were presenting NLF of great or mild importance due to ageing. We also performed on patients with a great importance NLF a lipofilling of the cheek and the lips in order to recover the volumes lost with ageing. The evaluation items were the quality of the filling (importance and symmetry), quality of the scars and complications occurring on the donor and receiving sites. RESULTS: Objective and subjective results were good. Projection of the NLF was in most of cases strongly and durably restored. CONCLUSION: Filling of the deep nasolabial fold using a temporalis superficialis fascia graft is a simple and reliable technique. Depending on the patients this method can be associated with a lipostructure of the cheek and the lips in order to optimize the facial rejuvenation. According to us this tissular implant allows a long lasting and important correction of the NLF.

Synchrotron radiation Fourier-transform infrared and Raman microspectroscopy study showing an increased frequency of creatine inclusions in the rat hippocampal formation following pilocarpine-induced seizures.

In the present work, synchrotron radiation Fourier-transform infrared (SRFTIR) and Raman microspectroscopies were used to evaluate a possible role of creatine in the pathogenesis and progress of pilocarpine-evoked seizures and seizure-induced neurodegenerative changes in the rat hippocampal tissue. The main goal of this study was to identify creatine deposits within the examined brain area, to analyze their frequency in epileptic animals and naive controls and to examine correlations between the number of inclusions in the hippocampal formation of epileptic rats and the quantitative parameters describing animal behavior during 6-h observation period after pilocarpine injection. The presence of creatine in the brain tissue was confirmed based on the vibrational bands specific for this compound in the infrared and Raman spectra. These were the bands occurring at the wavenumbers around 2800, 1621, 1398, and 1304 cm(-1) in IR spectra and around 1056, 908 and 834 cm(-1) in the Raman spectra. Creatine was detected in eight of ten analyzed epileptic samples and in only one of six controls under the study. The number of deposits in epileptic animals varied from 1 to 100 and a relative majority of inclusions were detected in the area of the Dentate Gyrus and in the multiform hippocampal layer. Moreover, the number of creatine inclusions was positively correlated with the total time of seizure activity.

[Hammer hypothenar syndrome: review of the literature and case report]. / Le syndrome du marteau hypothénarien: analyse de la littérature et cas clinique.

Hypothenar hammer syndrome is a rare disease first described by Conn et al. in 1970. It groups together symptoms of chronic microtraumatism to the ulnar artery or its superficial palmar branch against the hamate. Manual workers using vibrating tools are the most affected by this. Diagnosis is made by echodoppler, while arteriography is the gold standard for establishing the treatment plan. Hypothenar hammer syndrome may lead to severe complications secondary to ischemia and to embolic events resulting from delayed diagnosis or maltreatment. There is no real consensus as to the place of medical or surgical treatments. Medical treatment consists of eliminating favorizing factors and long-term antiplatelet aggregation treatment. Surgical treatment depends on the vascular lesions: simple arterial ligation, resection of the thrombosed arterial segment and end-to-end anastomosis, or revascularization using a pontage venous or arterial graft. Some authors suggested an associated thoracic sympathectomy. The diagnosis must be made early; the choice of treatment must be targeted at preventing serious embolic complications.

[Primary essential cutis verticis gyrata: Case report and literature review]. / Cutis verticis gyrata primitif essentiel : cas clinique et revue de la littérature.

INTRODUCTION: Cutis verticis gyrata (CVG) is a rare and slowly progressive deformity of the scalp with thick gyrated skin folds and ridges which are similar to gyri of the brain cortex. Those folds can lead to local skin infections, to a social and cosmetic complain. CVG can be classified into two forms: primary (essential and non-essential) and secondary. To date, fifteen operated cases of primary essential CVG have been reported in the medical literature. CASE REPORT: We report the case of an 18 year-old male patient with a primary essential CVG. There were several large skin folds in the sagittal axis on the vertex region, and in the coronal axis on the occipital region. He did not present any cutaneous complication. His main complains was the unaesthetic aspect of his scalp with a psychological complex. The disease had occurred during puberty. We present the excision pattern and the results with a six months follow-up. CONCLUSION: CVG can be treated surgically with resection of the thickened excess skin in coronal and sagittal axis. Scalp lift must be effective all over the different areas of the scalp. The scalp flaps must have a reliable vascularisation. Combined incisions of the galea help to treat the residual folds. The excision pattern must be reproductible, as this disease is progressive.

Estimating and correcting mie scattering in synchrotron-based microscopic fourier transform infrared spectra by extended multiplicative signal correction.

We present an approach for estimating and correcting Mie scattering occurring in infrared spectra of single cells, at diffraction limited probe size, as in synchrotron based microscopy. The Mie scattering is modeled by extended multiplicative signal correction (EMSC) and subtracted from the vibrational absorption. Because the Mie scattering depends non-linearly on alpha, the product of the radius and the refractive index of the medium/sphere causing it, a new method was developed for estimating the Mie scattering by EMSC for unknown radius and refractive index of the Mie scatterer. The theoretically expected Mie contributions for a range of different alpha values were computed according to the formulae developed by Van de Hulst (1957). The many simulated spectra were then summarized by a six-dimensional subspace model by principal component analysis (PCA). This subspace model was used in EMSC to estimate and correct for Mie scattering, as well as other additive and multiplicative interference effects. The approach was applied to a set of Fourier transform infrared (FT-IR) absorbance spectra measured for individual lung cancer cells in order to remove unwanted interferences and to estimate ranges of important alpha values for each spectrum. The results indicate that several cell components may contribute to the Mie scattering.