Evaluation of SP3 for antibody-free quantification of tau in CSF mimic and brain by mass spectrometry.

Tubulin-associated unit (tau) has an important role in the pathogenesis and the diagnosis of Alzheimer's disease (AD) and other tauopathies. In view of the diversity of tau proteoforms, antibody-free methods represent a good approach for unbiased quantification. We adapted and evaluated the single-pot, solid-phase-enhanced sample-preparation (SP3) protocol for antibody-free extraction of the tau protein in cerebro-spinal fluid (CSF) mimic and in human brain. A total of 13 non-modified peptides were quantified by high-resolution mass spectrometry (HRMS) after digestion of tau by trypsin. We significantly improved the basic SP3 protocol by carefully optimizing the organic solvents and incubation time for tau binding, as well as the digestion step for the release directly from the SP3 beads of the 13 tau peptides. These optimizations proved to be primarily beneficial for the most hydrophilic tau peptides, increasing the sequence coverage of recombinant tau. Mean recovery in CSF mimic of the 13 non-modified peptides was of 53%, with LODs ranging from 0.75 to 10 ng/mL. Next, we tested the optimized SP3 protocol on pathological tau extracted from the soluble fraction from an AD brain sample (middle frontal gyrus). We could successfully identify and quantify biologically relevant tau peptides including representative peptides of two isoforms and two phospho-peptides (pTau217 and pTau181).

Transition-Metal-Free Dehydropolymerization of Phosphine-Boranes at Ambient Temperature.

Stoichiometric reaction of phosphine-borane adducts RR'PH⋅BH3 (R=Ph, R'=H, Ph, Et, and R=R'=t Bu) with the strong acid HNTf2 (Tf=SO2 CF3 ) leads to H2 elimination and the formation of the triflimido derivatives, RR'PH⋅BH2 (NTf2 ). Subsequent deprotonation by using bases, such as diisopropylethylamine or the carbene IPr (IPr=N,N'-bis(2,6-diisopropylphenyl)imidazol-2-ylidene), led to the formation of P-mono- or -disubstituted polyphosphinoboranes [RR'P-BH2 ]n . Evidence for the intermediacy of transient phosphinoborane monomers, RR'PBH2 , was provided by trapping reactions.

Laryngeal paralysis secondary to cervical bite injuries in five dogs.

CASE HISTORIES: Medical records of a veterinary hospital in Belgium were reviewed for dogs (n = 5) that presented between 2016 and 2019 with laryngeal paralysis secondary to bite wounds to the cervical region received while fighting with other dogs. The time elapsed between the trauma and presentation was from a few hours up to 5 days. CLINICAL FINDINGS AND TREATMENT: Bilateral laryngeal paralysis was identified in three dogs and unilateral laryngeal paralysis in two dogs via endoscopic assessment of laryngeal function. The primary concomitant lesions included tracheal injury in 3/5 dogs and oesophageal injury in 1/5 dogs. One dog with bilateral laryngeal paralysis was treated medically as no signs of dyspnoea were present. Surgical management was elected in 4/5 dogs based on evaluation of their clinical status and lesions revealed by endoscopic examination of upper gastrointestinal and respiratory tracts. Dogs underwent surgical procedures that were determined to be appropriate for treatment of the lesions identified on clinical examination, diagnostic imaging, and endoscopy. The cervical region was explored through a ventral midline approach in 2/4 cases, to close tracheal perforations. Temporary tracheostomy was performed in 2/4 cases. Procedures to correct brachycephalic airway obstructive syndrome were performed in 2/4 cases. Cricoarytenoid lateralisation was performed in 2/4 dogs. Dogs were hospitalised for 2-10 days and received antimicrobial therapy before surgery and for 2-3 weeks after surgery. Physical examination and respiratory function were normal in 3/5 dogs 4-6 months after discharge. Information regarding outcomes for two cases was obtained from the owners by telephone assessment 1-6 months after surgery. The owner of each dog reported the respiratory function to be excellent. DIAGNOSIS: Uni- or bilateral, transient or permanent laryngeal paralysis with concomitant oesophageal, tracheal, or laryngeal lesions following cervical dog bite injuries diagnosed by endoscopic examination of upper gastrointestinal and respiratory tracts. CLINICAL RELEVANCE: This case series describes the diagnosis and management of dogs with laryngeal paralysis secondary to cervical dog bite injuries. To the authors' knowledge, this is the first published report documenting bilateral laryngeal paralysis secondary to cervical dog bite injuries. Clinicians should be aware of this pathology and the importance of investigating laryngeal function in dogs presenting with cervical bites, particularly those with inspiratory dyspnoea. Upper airway and digestive endoscopy are recommended for complete assessment of cervical traumatic injuries.

Impact of non-steroidal anti-inflammatory drugs on the efficiency of enhanced recovery programmes after colorectal surgery: a retrospective study of the GRACE database.

BACKGROUND: Multimodal analgesia is considered a key element of enhanced recovery programmes (ERPs) after colorectal surgery. We investigated the effects of NSAIDs, a major component of multimodal analgesia on adherence to ERP, incidence of postoperative complications, and length of hospital stay (LOS). METHODS: This was a retrospective study of the GRACE database that included 8611 patients scheduled for colorectal surgery with an ERP between February 2016 and November 2019. Primary endpoints were adherence to the postoperative protocol, the rate and type of postoperative complications, and LOS. Data are median [IQR] and number (per cent). Multivariate models were used to assess the effects of NSAIDs on these variables taking into account potential confounding factors. RESULTS: Data from 8258 patients were analysed and classified into four groups according to whether NSAIDs had been given intra- and/or postoperatively or not at all; 4578 patients were given NSAIDs intra- and/or postoperatively and 3680 patients received no NSAIDs. Use of NSAIDs was significantly (P<0.001) associated with improved adherence to the postoperative protocol (4.0 [3.0-4.0] vs. 3.0 [2.0-4.0] items), a reduced incidence of complications (21.1% vs. 29.2%), and a shortened LOS (5.0 [3.0-7.0] vs. 6.0 [4.0-9.0] days) compared to the no-NSAIDs group. Multivariate analyses adjusted for the confounding factors confirmed a significant (P<0.001) beneficial impact of NSAIDs on these three primary endpoints. CONCLUSION: This study suggests that perioperative NSAID use results in better adherence to the postoperative protocol, fewer postoperative in-hospital complications, and shorter LOS after colorectal surgery.

[Perception by the oncologist of the parental distress in childhood cancer survivor]. / Perception par l'oncologue de la détresse des parents d'un enfant en rémission de cancer.

The aim of the study was to assess the degree of similarity between the distress expressed by parents of a child cancer survivor and the distress perceived by the oncologist. Overall, 61 parents of a child cancer survivor (between 4 and 6 years of cancer remission) were recruited in Belgian hospitals (Province of Liège). Parents filled out questionnaires about the intolerance of uncertainty (IUS), positive beliefs about worry (WW-II), parental worries about the evolution of their child's health (QIPS-R15), problem orientation (NPOQ), cognitive avoidance (CAQ), ruminations (Mini-CERTS), and anxiety/depression (HADS). Eight oncologists who having taken care of the child filled out the «OncoMed¼ questionnaire that examines their perception of the parental distress (e.g. anxiety, worries). Parents suffered from anxiety,worries about their child's health and showed a low level of tolerance of uncertainty. Overall, oncologists demonstrated a low perceived distress compared to the level of parental distress. The study highlights the need to detect early parents who are intolerant of uncertainty in order to offer them an efficient psychological follow-up. This study underlines finally the necessity to develop medical perception tools about the distress in order to strengthen the medical communication and the follow-up of these families.

L'objectif de l'étude est de mesurer le degré de concordance entre la détresse exprimée par les parents d'un enfant en rémission de cancer et la détresse perçue par l'oncologue. Au total, 61 parents d'un enfant en rémission de cancer (de 4 années à 6 années de rémission) sont recrutés dans les hôpitaux belges (Province de Liège). Les parents remplissent des questionnaires relatifs à l'intolérance à l'incertitude (QII), aux croyances positives à l'égard de l'inquiétude (PSI-II), aux inquiétudes relatives à l'évolution de la santé de l'enfant (QIPS-R15), à l'orientation de l'attitude face à un problème (QAP), à l'évitement cognitif (QEC), aux ruminations (Mini-CERTS) et à l'anxiété/dépression (HADS). Huit oncologues ayant pris en charge l'enfant répondent au questionnaire «OncoMed¼ évaluant leur perception de la détresse du parent (e.g. anxiété, inquiétudes). Les parents souffrent d'anxiété, d'inquiétudes relatives à la santé de leur enfant et ont une faible tolérance à l'incertitude. Les oncologues perçoivent en moyenne, une détresse moins élevée que celle rapportée par les parents. L'étude souligne l'importance de détecter, dès la prise en charge de l'enfant, le facteur d'intolérance à l'incertitude chez le parent afin de lui proposer un suivi psychologique adéquat. Cette étude souligne enfin la nécessité de développer des outils de perception médicale de la détresse afin de renforcer la communication médicale et le suivi de ces familles.

[Experimental study on the intolerance of uncertainty and cognitive biases in parents of child cancer survivor]. / Étude expérimentale sur l'intolérance à l'incertitude et biais cognitifs chez les parents d'un enfant survivant de cancer.

Studies in pediatric oncology describe a relative good quality of life in child cancer survivor. However, few studies were interested in the parents of a child cancer survivor. 61 parents are recruited in the Belgian hospitals. Three groups of parents are constituted : the parents whose child is in 4 years of survivorship (group 1), in 5 years of survivorship (group 2) and in 6 years of survivorship (group 3). Clinical scales and a Stroop emotion task are administered. Parents (of the 3 groups) present a low tolerance of uncertainty, have excessive worries about the evolution of the health of their child, and suffer from anxious symptoms. The Stroop emotion tasks reveals a cognitive bias of the attention in favour of threatening stimuli. The study highlights the importance to detect parents who are intolerant of uncertainty at the cancer diagnosis stage and their continuous psychological follow-up once the treatments are ended.

Les études en oncologie pédiatrique décrivent une relativement bonne qualité de vie chez les enfants survivants de cancer. À ce jour, peu d'études se sont intéressées aux parents d'un enfant survivant de cancer. Soixante-et-un parents sont recrutés dans les hôpitaux belges. Trois groupes de parents sont constitués : les parents dont l'enfant est à 4 ans de rémission (groupe 1), à 5 ans de rémission (groupe 2) et à 6 ans de rémission (groupe 3). Des échelles cliniques et une tâche de Stroop émotion sont administrées. Les parents (des 3 groupes) présentent une faible tolérance à l'incertitude, ont des inquiétudes excessives quant à l'évolution de la santé de leur enfant et souffrent de symptômes anxieux. Le Stroop émotion révèle un biais cognitif de l'attention en faveur des stimuli de nature menaçante. L'étude met en exergue l'importance de détecter les parents intolérants à l'incertitude lors du diagnostic d'annonce du cancer et leur suivi psychologique continu une fois les traitements terminés.

Prevalence, location and concurrent diseases of ultrasonographic cyst-like lesions of abdominal lymph nodes in dogs.

Lymph nodal cyst-like lesions are occasionally identified during abdominal ultrasound in dogs. However, a study evaluating their prevalence and clinical significance is lacking. The aim of this observational cross-sectional study was to evaluate prevalence, most common location and concurrent diseases of cyst-like lymph nodes detected during abdominal ultrasound. Affected lymph nodes, patient signalment and concurrent diseases of dogs with cyst-like lymph nodal lesions having undergone abdominal ultrasound over a one-year period were recorded. Twenty-three affected lymph nodes were observed in 17/553 dogs (prevalence=3 per cent). The most commonly affected was the lumbar lymphocenter (7/23), followed by the coeliac (6/23), the cranial mesenteric (5/23) and the iliosacral (5/23). Twenty-three concurrent diseases were diagnosed in 17 dogs, among which 16/23 were non-neoplastic (70 per cent). The most common concurrent disease was renal insufficiency (8/23), followed by neoplasia (7/23), gastroenteropathy (3/23), benign prostatic disease (2/23), pancreatitis (1/23), peritonitis (1/23) and neurological disease (1/23). No statistical correlation existed between cyst-like lymph nodal lesion and a specific neoplastic or non-neoplastic disease. In conclusion, in the present study, cyst-like lymph nodal lesions have a low prevalence, involve different lymphocenters and were found in dogs affected by different diseases, including both non-neoplastic and neoplastic aetiologies.

Activation of Si-H bonds across the nickel carbene bond in electron rich nickel PC(carbene)P pincer complexes.

Silicon-hydrogen bonds are shown to add to a nickel carbon double bond to yield nickel α-silylalkyl hydrido complexes. Kinetic and isotope labeling studies suggest that a concerted 4-centred addition across the Ni=C bond is operative rather than a mechanism involving Si-H oxidative addition. This constitutes an example of Si-H bond activation via ligand cooperativity.

[Quality of life in children with cancer history and parental adaptation]. / Qualité de vie de l'enfant avec antécédents oncologiques et adaptation parentale.

This study assesses quality of life (Qol) in children with cancer history as well as Qol of their parents and examines the relationship between parental adjustment and children Qol. Two groups were formed: an experimental group composed by children with cancer antecedents and their parents and a control group with healthy children and their parents. Children have filled a questionnaire about their Qol. Parents have completed questionnaires about their coping, their illness perceptions, their parenting stress and their Qol. The results show that Qol was satisfactory and similar in both groups. Significant correlations were found between parental illness representations (threat and personal control) and children Qol as well as between coping strategies based on maintaining family cohesion and children Qol. This study highlights the need to assess the adjustment of parents after child cancer and to develop interventions targeting parental representations and coping strategies.

Is it possible to improve the breaking bad news skills of residents when a relative is present? A randomised study.

BACKGROUND: Although patients with cancer are often accompanied by a relative during breaking bad news (BBN) consultations, little is known regarding the efficacy of training programmes designed to teach residents the communication skills needed to break bad news in a triadic consultation. METHODS: Residents were randomly assigned to a 40-h dyadic and triadic communication skills training programme (n=48) or a waiting list (n=47). A simulated BBN triadic consultation was audiotaped at baseline, and after training for the training group, and 8 months after baseline for the waiting list group. Transcripts were analysed using content analysis software (LaComm). A coder determined the moment of bad news delivery and the relative's first turn of speech regarding the bad news. A generalised estimating equation was used to evaluate residents' communication skills, BBN timing, and the relative's inclusion in the consultation. RESULTS: Ninety-five residents were included. After training, the duration of the pre-delivery phase was found to be longer for the trained residents (relative risk (RR)=3.04; P<0.001). The simulated relative's first turn of speech about the bad news came more often during the pre-delivery phase (RR=6.68; P=0.008), and was more often initiated by the trained residents (RR=19.17; P<0.001). Trained residents also used more assessment (RR=1.83; P<0.001) and supportive utterances (RR=1.58; P<0.001). CONCLUSION: This study demonstrates that a training programme that focuses on the practice of dyadic and triadic communication skills can improve the communication skills of the participating residents in a BBN triadic consultation. Such a training should be included in resident curriculum.

Interstitial laser photocoagulation for the treatment of osteoid osteoma: retrospective study on 35 cases.

PURPOSE: The aim of our retrospective study was to evaluate the efficacy of interstitial laser photocoagulation for the treatment of osteoid osteomas and to identify the factors leading to failure of the procedure. MATERIAL AND METHODS: Thirty-five patients received interstitial laser photocoagulation treatment. A minimum of 3 months follow-up was required. The laser fibre was positioned within the nidus under CT guidance, and energy between 500 and 1800 J was delivered at a power of 2 Watts/s. Follow-up was by outpatient appointment and through a questionnaire sent to the patients. RESULTS: Thirty patients replied to the questionnaire. Mean follow-up was 40 months. The technical success rate was 100%. The primary success rate (no pain at 1 month) was 94.3%. The rate of recurrence was 6%. The rate of complications was 11.4% (a skin burn, patellar enthesopathy, a broken item of material, sacro-iliac fibrous alterations). Factors likely to favour failure of the procedure are the size of the nidus (P=0.04) and poor positioning of the laser fibre (P=0.03). CONCLUSION: Interstitial laser photocoagulation is an effective and safe treatment for osteoid osteomas.

The increase from 2.5 to 5 mg/kg of rabbit anti-thymocyte-globulin dose in reduced intensity conditioning reduces acute and chronic GVHD for patients with myeloid malignancies undergoing allo-SCT.

We previously reported that reduced intensity conditioning (RIC) regimen with fludarabine, BU and 2.5 mg/kg of rabbit anti-thymocyte globulin (r-ATG) was effective but associated with a high rate of acute and chronic GVHD. Therefore, we increased the dose of r-ATG to 5 mg/kg. In this report, we analyzed 87 patients with AML or myelodysplastic syndrome (MDS) undergoing allo-SCT from an HLA-identical sibling donor from 2000 to 2010. RIC consisted of fludarabine, BU and r-ATG 2.5 mg/kg on 1 day (r-ATG1; n=53) or 2.5 mg/kg per day over 2 days (r-ATG2; n=22). Grade 2-4 acute GVHD incidence at day 100 was 30.2% and 8.8% in the r-ATG1 and r-ATG2 groups, respectively (P=0.038). Extensive chronic GVHD incidence was 60.4% and 12% in the r-ATG1 and r-ATG2 groups, respectively (P<0.001). The relapse incidences (RI) at 24 months were 18.9% and 28.5% in r-ATG1 and r-ATG2 groups, respectively (P=0.640). Overall and PFS were not different between the r-ATG1 and r-ATG2 groups. r-ATG dose at 5 mg/kg in the setting of RIC seems a good balance allowing GVHD prevention and antitumor effect with a remarkable reduction of GVHD incidence without an identical level of increased relapse rate.

Quality of work life in doctors working with cancer patients.

BACKGROUND: Although studies have shown that medical residents experience poor psychological health and poor organizational conditions, their quality of work life (QWL) had not been measured. A new tool, the Quality of Work Life Systemic Inventory (QWLSI), proposes to fill the gap in the definition and assessment of this concept. AIMS: To confirm the convergent validity of the QWLSI, analyse Belgian medical residents' QWL with the QWLSI and discuss an intervention methodology based on the analysis of the QWLSI. METHODS: One hundred and thirteen medical residents participated between 2002 and 2006. They completed the QWLSI, the Maslach Burnout Inventory and the Job Stress Survey to confirm the correspondence between these three tools. RESULTS: Residents' low QWL predicted high emotional exhaustion (ß = 0.282; P < 0.01) and job stress (ß = 0.370; P < 0.001) levels, confirming the convergent validity. This sample of medical residents had an average QWL (µ = 5.8; SD = 3.1). However, their QWL was very low for three subscales: arrangement of work schedule (µ = 9; SD = 6.3), support offered to employee (µ = 7.6; SD = 6.1) and working relationship with superiors (µ = 6.9; SD = 5.3). CONCLUSIONS: The results confirm that the QWLSI can provide an indication of workers' health well-being and of organizational performance in different areas of work life. The problem factors found among Belgian medical residents suggest that prevention should focus on reduction of work hours, development of support and change in leadership style.

Predictors of early outcome after acute appendicitis: is delaying surgery for acute appendicitis an option? A retrospective study.

PURPOSE: This study analysed the clinical and para-clinical criteria that may allow surgeons and emergency physicians to take a decision regarding the surgery of acute appendicitis. METHODS: A retrospective analysis was conducted on 284 acute appendicitis patients who underwent surgery between January 2007 and December 2009 in our institution. The registered data were extracted from patient files and statistically analysed. These data included past medical history, clinical, laboratory and imaging data, duration of hospital stay and post-operative complications. Patient delay (time between the appearance of symptoms and patient arrival at the emergency department) and hospital delay (time between hospital arrival and operation) were correctly investigated. Statistical analysis was done by using SPSS software. RESULTS: The patient delay is significantly increased in relation to the severity of appendicitis: 24 h (10.8-30.8 h) versus 37.4 h (36.8-38 h) (P < 0.05), unlike hospital delay, which remains constant (between simple and severe appendicitis): 7.5 h (5-14.8 h) versus 8 h (5-13 h). In severe appendicitis, the proportion of guarding, rebound tenderness, tachycardia (P < 0.05) and fever (P < 0.005) were significantly high, and leucocytosis (P < 0.05), C-reactive protein (CRP) (P < 0.001) and eosinopaenia [37.0 vs. 72.8 (P < 0.001)] were significantly different. Concerning computed tomography (CT) and echography, perforation, abscess formation (P < 0.05), phlegmon (P < 0.005) and peritonitis (P < 0.05) were significant signs of complicated cases. The length of hospital stay (P < 0.001) and duration of antibiotic therapy (P < 0.001) were statistically significant in cases of complicated appendicitis. CONCLUSION: Patient delay is a determining factor for the grade of appendicitis. It has an influence on the complications, length of hospital stay and duration of antibiotic treatment, unlike hospital delay.

Critically ill cancer patients in the intensive care unit: short-term outcome and 1-year mortality.

BACKGROUND: The short-term survival of critically ill patients with cancer has improved over time. Studies providing long-term outcome for these patients are scarce. METHODS: We prospectively analyzed outcomes and rates of successful discharge of 111 consecutive critically ill cancer patients admitted to intensive care unit (ICU) in 2008 and identified factors influencing these results. RESULTS: ICU mortality was 32% and hospital mortality was 41%. None of the characteristics of the malignancy nor age or neutropenia were significantly different between survivors and others. Two variables were independently associated with ICU mortality: high Logistic Organ Dysfunction score on day 7 and a diagnosis of viral infection and/or reactivation. The 1-year mortality rate for ICU survivors was 58% and was significantly lower in patients with a diagnosis of acute leukemia or multiple myeloma. CONCLUSION: Organ failure scores on day 7 can predict outcome for cancer patients in the ICU. Viral infection and reactivation appear to worsen the prognosis. One-year mortality rate is high and depends on the malignancy.

Comparison of radiation-induced segregation in ultrafine-grained and conventional 316 austenitic stainless steels.

Due to a high number density of grain boundaries acting as point defect sinks, ultrafine-grained materials are expected to be more resistant to irradiation damage. In this context, ultrafine-grained 316 austenitic stainless steel samples have been fabricated by high pressure torsion. Their behavior under ion irradiation has been studied using atom probe tomography. Results are compared with those obtained in an ion irradiated conventional coarse-grained steel. The comparison shows that the effects of irradiation are limited and that intragranular and intergranular features are smaller in the ultrafine-grained alloy. Using cluster dynamic modeling, results are interpreted by a higher annihilation of point defects at grain boundaries in the ultrafine-grained steel.

Is it possible to improve residents breaking bad news skills? A randomised study assessing the efficacy of a communication skills training program.

BACKGROUND: This study aims to assess the efficacy of a 40-h training programme designed to teach residents the communication skills needed to break the bad news. METHODS: Residents were randomly assigned to the training programme or to a waiting list. A simulated patient breaking bad news (BBN) consultation was audiotaped at baseline and after training in the training group and 8 months after baseline in the waiting-list group. Transcripts were analysed by tagging the used communication skills with a content analysis software (LaComm) and by tagging the phases of bad news delivery: pre-delivery, delivery and post-delivery. Training effects were tested with generalised estimating equation (GEE) and multivariate analysis of variance (MANOVA). RESULTS: The trained residents (n=50) used effective communication skills more often than the untrained residents (n=48): more open questions (relative rate (RR)=5.79; P<0.001), open directive questions (RR=1.71; P=0.003) and empathy (RR=4.50; P=0.017) and less information transmission (RR=0.72; P=0.001). The pre-delivery phase was longer for the trained (1 min 53 s at baseline and 3 min 55 s after training) compared with the untrained residents (2 min 7 s at baseline and 1 min 46 s at second assessment time; P<0.001). CONCLUSION: This study shows the efficacy of training programme designed to improve residents' BBN skills. The way residents break bad news may thus be improved.

A biochemical and functional protein complex involving dopamine synthesis and transport into synaptic vesicles.

Synaptic transmission depends on neurotransmitter pools stored within vesicles that undergo regulated exocytosis. In the brain, the vesicular monoamine transporter-2 (VMAT(2)) is responsible for the loading of dopamine (DA) and other monoamines into synaptic vesicles. Prior to storage within vesicles, DA synthesis occurs at the synaptic terminal in a two-step enzymatic process. First, the rate-limiting enzyme tyrosine hydroxylase (TH) converts tyrosine to di-OH-phenylalanine. Aromatic amino acid decarboxylase (AADC) then converts di-OH-phenylalanine into DA. Here, we provide evidence that VMAT(2) physically and functionally interacts with the enzymes responsible for DA synthesis. In rat striata, TH and AADC co-immunoprecipitate with VMAT(2), whereas in PC 12 cells, TH co-immunoprecipitates with the closely related VMAT(1) and with overexpressed VMAT(2). GST pull-down assays further identified three cytosolic domains of VMAT(2) involved in the interaction with TH and AADC. Furthermore, in vitro binding assays demonstrated that TH directly interacts with VMAT(2). Additionally, using fractionation and immunoisolation approaches, we demonstrate that TH and AADC associate with VMAT(2)-containing synaptic vesicles from rat brain. These vesicles exhibited specific TH activity. Finally, the coupling between synthesis and transport of DA into vesicles was impaired in the presence of fragments involved in the VMAT(2)/TH/AADC interaction. Taken together, our results indicate that DA synthesis can occur at the synaptic vesicle membrane, where it is physically and functionally coupled to VMAT(2)-mediated transport into vesicles.

Antiproliferative and antimalarial anthraquinones of Scutia myrtina from the Madagascar forest.

Bioassay-guided fractionation of an ethanol extract of a Madagascar collection of the bark of Scutia myrtina led to the isolation of three new anthrone-anthraquinones, scutianthraquinones A, B and C (1-3), one new bisanthrone-anthraquinone, scutianthraquinone D (4), and the known anthraquinone, aloesaponarin I (5). The structures of all compounds were determined using a combination of 1D and 2D NMR experiments, including COSY, TOCSY, HSQC, HMBC, and ROESY sequences, and mass spectrometry. All the isolated compounds were tested against the A2780 human ovarian cancer cell line for antiproliferative activities, and against the chloroquine-resistant Plasmodium falciparum strains Dd2 and FCM29 for antiplasmodial activities. Compounds 1, 2 and 4 showed weak antiproliferative activities against the A2780 ovarian cancer cell line, while compounds 1-4 exhibited moderate antiplasmodial activities against P. falciparum Dd2 and compounds 1, 2, and 4 exhibited moderate antiplasmodial activities against P. falciparum FCM29.

Myelofibrosis-associated lymphoproliferative disease: retrospective study of 16 cases and literature review.

Background. To better describe the clinical, biological, and the outcome of non-Hodgkin's lymphoma (NHL) with, at the initial presentation, bone marrow fibrosis (MF). Patients and Methods. From January 2001 to January 2007, 16 eligible patients with NHL and MF were retrieved from the Pathology Department of the University hospital of Amiens. Median age of patients was 62 years (range 16-74) with a sex ratio male/female of 3. Results. MF is associated with all types of lymphoma predominantly with B-cell phenotype and it seems to be more associated with low-grade NHL. B-symptoms are more frequent at diagnosis and more patients presented with an elevated LDH level. JAK-2 was negative in the 10 patients analysed. Two patients presented with features of primary MF with no evidence of lymphoma. Overall response rate was 94% after the first line of therapy with regression or improvement of MF. Relapse occurred in 8 patients (47%) with recurrence of MF in all of them. After a median follow-up of 42 months, 12 patients were alive with an overall survival rate for the entire group of 75%. Conclusions. MF-associated NHL is a rare manifestation which may be associated with all types of NHL and its presence does not seem to confer a poor prognosis. A search for lymphoproliferation should be considered when the cause of MF is not apparent.