HPLC & NMR-based forced degradation studies of ifosfamide: The potential of NMR in stability studies.

PURPOSE: The aim of this study is to conduct a forced degradation study on ifosfamide under several stress conditions to investigate the robustness of the developed HPLC method. It also aims to provide further insight into the stability of ifosfamide and its degradation profile using both HPLC and NMR. METHODS: Ifosfamide solutions (20mg/mL; n=15, 20mL) were stressed in triplicate by heating (70°C), under acidic (pH 1 & 4) and alkaline (pH 10 & 12) conditions. Samples were analysed periodically using HPLC and FT-NMR. RESULTS AND DISCUSSION: Ifosfamide was most stable under weakly acidic conditions (pH 4). NMR results suggested that the mechanism of ifosfamide degradation involves the cleavage of the PN bond. For all stress conditions, HPLC was not able to detect ifosfamide degradation products that were detected by NMR. CONCLUSION: These results suggest that the developed HPLC method for ifosfamide did not detect the degradation products shown by NMR. It is possible that degradation products co-elute with ifosfamide, do not elute altogether or are not amenable to the detection method employed. Therefore, investigation of ifosfamide stability requires additional techniques that do not suffer from the aforementioned shortcomings.

Neurologic Disorders in Immunocompetent Patients with Autochthonous Acute Hepatitis E.

Neurologic disorders, mainly Guillain-Barré syndrome and Parsonage­Turner syndrome (PTS), have been described in patients with hepatitis E virus (HEV) infection in industrialized and developing countries. We report a wider range of neurologic disorders in nonimmunocompromised patients with acute HEV infection. Data from 15 French immunocompetent patients with acute HEV infection and neurologic disorders were retrospectively recorded from January 2006 through June 2013. The disorders could be divided into 4 main entities: mononeuritis multiplex, PTS, meningoradiculitis, and acute demyelinating neuropathy. HEV infection was treated with ribavirin in 3 patients (for PTS or mononeuritis multiplex). One patient was treated with corticosteroids (for mononeuropathy multiplex), and 5 others received intravenous immunoglobulin (for PTS, meningoradiculitis, Guillain-Barré syndrome, or Miller Fisher syndrome). We conclude that pleiotropic neurologic disorders are seen in HEV-infected immunocompetent patients. Patients with acute neurologic manifestations and aminotransferase abnormalities should be screened for HEV infection.

[Mandibular metastasis of a cutaneous melanoma or metachronous amelanotic melanoma of the oral cavity? A case report and literature review]. / Métastase mandibulaire d'un mélanome cutané ou mélanome achromique primitif métachrone de la cavité orale ? À propos d'un cas et revue de la littérature.

Primary and metastatic mandibular melanoma are extremely rare. We report the original case of a 55-year-old woman treated 16 years before for a cutaneous melanoma, and now presenting with a huge mandibular amelanotic melanoma. Was it an histologically different mandibular metastasis of the previous cutaneous melanoma, or a metachronous oral amelanotic melanoma?

Gadolinium induced recurrent acute pancreatitis.

Acute pancreatitis is a sudden swelling and inflammation of the pancreas. The two most common causes are alcohol use and biliary stones. Drug-induced acute pancreatitis are rare (1.4-2%). In this present study, we present a case of recurrent acute pancreatitis induced by a specific magnetic-resonance-imaging (MRI) contrast agent called gadobenate dimeglumine.

[Buccal mucosal graft for the treatment of urethral stricture]. / Greffe de muqueuse jugale pour le traitement d'une sténose urétrale.

INTRODUCTION: The indication of the buccal mucosal graft for urethroplasty has evolved over recent years. The ease of its harvesting, availability, and immunohistological properties, as well as its satisfactory results, has made the buccal mucosal graft the current procedure of choice. We report a case of use of a buccal mucosal graft to treat an urethral stricture correction. CASE REPORT: A 48-year-old man underwent a buccal mucosal graft for post-infectious urethral stricture correction. An inner right cheek graft of 50 × 30 mm was harvested and inserted into place after complete excision of the stricture. Postoperative evolution was satisfactory with no pain at the 5th day, resumption of normal diet at the 12th day, a significant improvement of peak flow rate at the 21st day. Follow-up examination at the 7th week revealed a mouth opening to 40 mm with complete healing. DISCUSSION: Buccal mucosal graft has currently the highest success rate compared to other surgical techniques as full thickness skin graft from hair or graft of bladder mucosa. Its harvesting can be single or multiple, however care must be taken as regards Stenon's duct and to the labial commissure. The complications of the donor site are infrequent and can be characterized by numbness and limited mouth opening. Tissues that contain immunohistological properties, which are similar to those of the urothelium, buccal mucosal graft, have become the gold standard for this type of correction.

Influence of HCV genotype 1 subtypes on the virus response to PEG interferon alpha-2a plus ribavirin therapy.

New factors that influence the viral response in HCV non-genotype 2/3 patients must be identified in order to optimize anti-HCV treatment. This multicenter prospective study evaluates the influence of HCV variability and pharmacological parameters on the virological response of these patients to pegylated interferon α2a (peg-IFN-α2a: 180 µg/week) and ribavirin (RBV; 800-1,200 mg/day) for 48 weeks. HCV subtypes were identified by sequencing the NS5B region. Serum RBV and peg-IFN-α2a concentrations were measured at weeks 4 and 12. The 115 patients (67 men; median age = 49, range 31-76) included 64 who had never been treated and 27 co-infected with HIV. The mean baseline HCV RNA was 6.30 ± 0.06 log IU/ml and the HCV genotypes were: G1 (n = 93) with 1a (n = 37) and 1b (n = 50), G4 (n = 20) and G5 (n = 2). Most patients (79/108; 73%) had an early virological response. Independent predictors of an early virological response were interferon naive patients (OR= 2.98, 95% CI: 1.15-7.72) and RBV of >2,200 ng/ml at week 12 (OR = 3.41, 95% CI: 1.31-8.90). Forty of 104 patients (38%) had a sustained virological response. The only independent predictors of a sustained virological response were subtype 1b (OR = 6.82, 95% CI: 1.7-26.8), and HCV RNA <15 IU/ml at week 12 (OR = 25, 95% CI: 6.4-97.6). Thus a serum RBV concentration of >2,200 ng/ml was associated with an early virological response and patients infected with HCV subtype 1b had a better chance of a sustained virological response than did those infected with subtype 1a.

[An update on focal infection of oral origin]. / Point actuel sur l'infection focale d'origine buccodentaire.

Focal infection of oral origin means that an oral infectious focus may have widespread effects. This concept remains controversial since it is difficult to prove the oral origin of germs responsible for an extra-oral infection. Experiments on animal models and clinical studies suggested several physiopathological mechanisms: bacteremia, toxinic and immunological mechanisms. Various operations induce the passage of bacterial flora (transcytosis) and its toxins into the bloodstream: oral care, chewing, or tooth brushing. Bacteremia is worsened by poor oral hygiene or an infection. The germs are usually destroyed by the host's reticuloendothelial system in a few minutes, but the presence of a valvular disease or a weak immune system favors focal infection. Besides infectious endocarditis, this may concern cardiovascular diseases, lung infections, prematurity and hypotrophy, diabetes, prosthetic infections, cerebral abscesses, etc. This update is based on literature review, selected according to its high level of scientific proof, as well as on a selected choice of consensus conferences. The current recommendation is to limit antibiotic prophylaxis to the high bacteremia risk procedures and to patients highly at risk of developing a focal infection.

CT scan study of the mandibular nerve intra-mandibular path.

INTRODUCTION: It is mandatory to know the anatomic path of the mandibular nerve and its intra-mandibular connections for numerous odontology, stomatology, and maxillofacial surgical procedures. We present a computed tomography study of the mandibular nerve intra-mandibular path. PATIENTS AND METHOD: The computed tomography of 14 mandibles was performed (vestibulo-lingual cross-section) and four landmarks were defined behind the mental foramen. At each landmark, we measured the distance between the alveolar canal and the two cortical layers in the vestibulo-lingual plane, and the distance between the alveolar canal and the alveolar crest, and the inferior border of the mandible in the vertical plane. RESULTS: In the vestibulo-lingual plane, the mandibular nerve is located close to the vestibular cortical layer, then to the lingual cortical before it exits. In the crestal plane, it is located at the superior third of the inferior border of the mandible. DISCUSSION: In spite of anatomical variability, there is a globally common path which means that the utmost caution should be taken by performing systematic imaging before undertaking any surgery close to the nerve.

[Upper aerodigestive tract erythroplakia]. / Les érythroplasies des voies aérodigestives supérieures.

Erythroplakia is a red mucosal macule with a chronic evolution. It is diagnosed after excluding traumatic, vascular, or inflammatory etiologies. Erythroplakia is rare in the upper aerodigestive tract. It affects middle-aged adults. The main predisposing factors are those of in situ carcinoma. Lesions are mainly located on lips or mucosa. Erythroleukoplakia may also occur. Ninety-one percent of erythroplakia are severe dysplasia, in situ carcinoma, or invasive carcinoma. Excision and histopathological examination are mandatory.

[Polymorphous low-grade adenocarcinoma of the maxilla simulating an odontogenic cyst]. / Adénocarcinome polymorphe de bas grade du maxillaire simulant un kyste odontogénique.

INTRODUCTION: In 1990, the WHO classified Polymorphous Low Grade Adenocarcinoma as a low-grade malignant tumor of the minor salivary glands. This tumor often originates from the hard or soft palate minor salivary glands. We report the first case revealed as an infected maxillary odontogenic cyst around an impacted tooth. CASE REPORT: A 50-year-old female presented 20 months ago with a bulging, painful oral mass in the right superior vestibule. The diagnosis was infected odontogenic cyst associated with an impacted canine tooth. A course of oral antibiotics was given and cyst enucleation and tooth extraction were performed 6 days later. Ameloblastoma was suggested on macroscopic findings; nevertheless, the pathologic end diagnosis was Polymorphic Low-Grade Adenocarcinoma of minor salivary gland origin. After negative carcinological screening, wide surgical resection was performed. Postoperative external radiotherapy was applied to the operated area as well as to cervical lymph node areas. DISCUSSION: Most of the 565 published cases concern a palatine location. Even though wide surgical resection is sufficient, external radiotherapy may be used on a case-by-case basis.

Liberal selection criteria for liver transplantation for hepatocellular carcinoma.

BACKGROUND: To help increase the number of transplants available for hepatocellular carcinoma in cirrhotic livers, this single-centre retrospective study compared the safety and feasibility of new, more liberal, selection criteria--no more than five tumours, with the largest tumour no greater than 5 cm (5/5 criteria)--with classical criteria. METHODS: Data from operations performed in 1990-2005 were extracted from preoperative radiological findings and postoperative specimen analyses, and four groups were constructed: Paul Brousse, Milan, University of California, San Francisco (UCSF) and 5/5 criteria. A fifth group comprised patients whose tumour load exceeded the 5/5 criteria. Survival and recurrence rates were compared. RESULTS: For the 110 patients in the study, survival rates (overall and disease-free) were 72.8 and 66.8 per cent at 5 and 10 years respectively, with a 5.5 per cent recurrence rate. The 5-year survival rate was 65, 77, 68 and 77 per cent for Paul Brousse, Milan, UCSF and 5/5 preoperative radiological criteria, with recurrence rates of 4, 4, 3 and 3 per cent, respectively. On multivariable analysis, the only factor that influenced survival was tumour load in excess of the 5/5 criteria. CONCLUSION: Use of the more liberal 5/5 criteria for selecting patients for liver transplantation results in similar disease-free and overall survival rates to classical criteria.

Acute hepatitis E in south-west France over a 5-year period.

BACKGROUND: Hepatitis E was found in people living in industrialized countries who had not travelled to highly endemic areas. OBJECTIVES: To study the cases of acute hepatitis E confirmed thanks to viral genomic detection over a 5 years period in south-west France. STUDY DESIGN: 62 cases of hepatitis E were identified between 2003 and 2007. Their demographic, clinical, and virological features were analyzed. RESULTS: Cases of acute hepatitis E occurred regularly throughout this period. No seasonal variation was found. Patients, usually male (sex ratio=1.95), were adults living in both urban and rural areas. Sixty (96.8%) patients had not travelled abroad during the 6 months before diagnosis. Clinical manifestations ranged from asymptomatic infection to severe hepatitis. HEV was genotyped in 55 specimens. All the patients who had not travelled abroad were infected with genotype 3. CONCLUSION: The incidence of hepatitis E in south-west France was stable from 2003 to 2007, 96.8% of the cases were autochthonous. There was an age-related increase in the disease and patients tended to be men. The predominant genotype and subtype was 3f. However, contaminations pathways involved in hepatitis E in our area remain to clarify.

[Mixed odontogenic tumours]. / Tumeurs odontogéniques mixtes.

Mixed odontogenic tumors include all tumors of the teeth both epithelial and mesenchymatous. Most of these tumors are rare (ameloblastic fibroma, ameloblastic fibro-odontoma and fibrodentinoma, odontoameloblastoma, calcified odontogenic cyst). Other tumors such as odontoma are frequent. They have in common a benign aspect, their often-fortuitous observation, a higher rate in the young adult and surgical treatment by complete exeresis preventing recurrence. They present a radiolucent lacuna but appear different depending on the presence or not of intralesional calcifications revealing the production of more or less mature dental tissue. The diagnosis may be confirmed only by an anatomopathological examination, along with clinical and radiological observations.

[Gingivoperiosteoplasty associated to bone graft: radiological evaluation]. / Gingivopériostoplasties associées à une greffe osseuse : évaluation radiologique.

INTRODUCTION: Gingivoperiosteoplasty associated to bone graft is part of a therapeutic strategy applied to the first 20 years of a patient's life. Management is pluridisciplinary. Most authors recommend a bone graft in mixed dentition at the end of premaxillary growth. Retroalveolar and panoramic radiography are the most often used to assess the bone height of the grafted site. We retrospectively studied the radiographies of 57 alveolar grafts in 44 patients. MATERIAL AND METHODS: Between 1999 and 2005, 44 patients underwent gingivoperiosteoplasty associated to bone graft. Thirteen underwent bilateral reconstruction. The surgical interventions were performed by the same surgeon. One year after surgery, the panoramic radiographies were analyzed by a single expert. The bone height compared to roots of adjacent teeth was classified in four grades. Grades 1 and 2 were considered as satisfactory or good and grades 3 and 4 not satisfactory and an indication for a new bone graft. In case of bilateral cleft, each side was analyzed independently. RESULTS: Grades 1 and 2 accounted for 84.2% of grafts. There was no statistical difference in alveolar bone height between patients presenting with agenesis of the lateral incisive. Eighty-one percent of patients grafted with mixed dentition (66% of the cases) had satisfactory results (35% of grade 1 and 46% of grade 2). Patients operated on after 15 years (n=15) had good results in 75% of the cases, 33% were bilateral cleft patients. There was no statistical difference between patients operated on early and those with delayed surgery. DISCUSSION: Radiological results for gingivoperiosteoplasty associated to bone graft are satisfactory. The procedure is easy, cheap, and reproducible. Evaluation with panoramic radiography is not as accurate as with the Denta Scan. CT scan is not used systematically to follow up alveolar cleft palate in children so as to limit irradiation. Volumetric tomography (cone beam) may be the best assessment.

KLF6 transcription factor protects hepatocellular carcinoma-derived cells from apoptosis.

Hepatocellular carcinoma (HCC) is a major public health concern because of the absence of early diagnosis and effective treatments. Efficient diagnosis modalities and therapies to treat HCC are needed. Kruppel-like factor (KLF) family members, such as KLF6, are involved in cell proliferation and differentiation. KLF6 is inactivated in solid tumors, which may contribute to pathogenesis. However, KLF6 status in HCC is controversial. Thus, we undertook the characterization of KLF6 expression and function in HCC and HCC-derived cell lines. We found that HCC, HepG2 and HuH7 cells expressed KLF6 messenger ribonucleic acid and protein. Next, using RNA interference, we demonstrated that inhibiting KLF6 expression in vitro strongly impaired cell proliferation-induced G1-phase arrest, inhibited cyclin-dependent kinase 4 and cyclin D1 expression, and subsequent retinoblastoma phosphorylation. Finally, KLF6 silencing caused p53 upregulation and inhibited Bcl-xL expression, to induce cell death by apoptosis. Taken together, these data demonstrated that KLF6 is essential for HCC-derived cells to evade apoptosis.

Expression of the transcription factor Klf6 in cirrhosis, macronodules, and hepatocellular carcinoma.

BACKGROUND AND AIMS: Macronodules (MN) occurring in cirrhosis are considered to be precursor lesions for hepatocellular carcinoma (HCC). However, early molecular events in hepatocellular carcinogenesis are poorly understood. The aim of this study was to compare gene expression profiling between cirrhotic tissues, MN, and HCC, to identify genes early involved in liver carcinogenesis. METHODS: Tissues were obtained from explanted livers: nine cirrhosis, 10 MN, and seven HCC. Total RNAs were extracted by RNeasy and reverse transcribed with labelled [(33)P]-alpha ATP. Hybridations were performed on Atlas Human Cancer 1.2 membranes (1176 genes). RESULTS: A two-way hierarchical clustering algorithm successfully isolated specific gene expression profiles when comparing MN, cirrhosis, and HCC. A total of 16 and 14 genes were up- and down-expressed, respectively, in HCC as compared to cirrhotic tissues. The molecular signature of MN was characterized by the down-expression of 23 and 42 genes as compared to cirrhosis and HCC, respectively. Among them, Klf6 was down-expressed in all MN samples whereas it was over-expressed in cirrhosis and HCC. This result was confirmed at RNA level by quantitative real time-polymerase chain reaction and at protein level by Western blotting. However, no mutation in the exon 2 of Klf6 was detected. CONCLUSION: We identified a molecular signature of MN characterized by a down-expression of several genes. One of them, Klf6 was found to be down-expressed in all MN without evidence of somatic mutations in the exon 2. This gene could be involved at an early stage of hepatocarcinogenesis.

[Stereotactic hypofractionated radiation therapy as a bridge to transplantation for hepatocellular carcinoma: Case report of a complete pathological response and review of the literature]. / Place de la radiothérapie en conditions stéréotaxiques en traitement d'attente du carcinome hépatocellulaire avant transplantation hépatique : à propos d'un cas de réponse histologique complète sur explant et revue de la littérature.

Patients with hepatocellular carcinoma who are on liver transplant waiting list usually require local treatment to limit any risk of tumour growth. Historically percutaneous radiofrequency ablation or transarterial chemoembolization represented the major therapeutic alternatives. Depending on the size, or the topography of the lesion these two techniques may not be feasible. Radiation therapy under stereotactic conditions has recently emerged in the management of localized hepatocellular carcinoma as an alternative to the focused therapies performed to date. We herein report the case of a 43-year-old patient harbouring a complete histological response on explant after liver stereotactic irradiation and discuss its role in the management of hepatocellular carcinoma before liver transplantation.

Hepatitis C virus viral load after conversion from tacrolimus to cyclosporine in liver transplant patients: a pilot study.

UNLABELLED: We assessed whether conversion from tacrolimus (TAC) to cyclosporine (CsA) was associated with a reduction in hepatitis C virus (HCV) viral load among HCV-positive liver transplant (OLT) patients. PATIENTS AND METHODS: Nine OLT patients with recurrent HCV have TAC and prednisone immunosuppression. None received any HCV antiviral therapy. After the last intake of TAC, the patients underwent a 12-hour area under the curve (AUC(12)) measurement of both TAC and HCV viral loads. The next morning (D(0)) patients were given CsA (4 mg/kg bid). At the first intake of CsA and at 1 month (M(1)) later, the patients underwent AUC(12) for CsA and HCV viral loads. Biological data, including aspartate (AST) and alanine (ALT) aminotransferase, gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (AP), and bilirubin levels, were collected during AUC(12), and at M(1) and M(3). RESULTS: With respect to liver enzymes (AST, ALT, GGT), there was no significant difference between D(0), M(1), and M(3). Conversely, there was a significant decrease in AP between D(0) and M(3) (P = .02), and a significant increase in total bilirubin between D(0) and M(1) (P = .04), and between D(0) and M(3) (P = .01). HCV viral load significantly increased by M(3) (P = .01). At no time (D(0), M(1)) was there any correlation between the AUC(12) of TAC or CsA, and between AUC(12) HCV viral load. CONCLUSION: This pilot study found no acute or chronic anti-HCV effects from CsA that were evident within 12 hours after CsA administrations or beyond 1 month of CsA therapy, respectively.