Organ donation by Maastricht-III pediatric patients: Recommendations of the Groupe Francophone de Réanimation et Urgences Pédiatriques (GFRUP) and Association des Anesthésistes Réanimateurs Pédiatriques d'Expression Française (ADARPEF). Part II: Specific organizational and technical considerations.

A panel of pediatric experts met to develop recommendations on the technical requirements specific to pediatric controlled donation after planned withdrawal of life-sustaining therapies (Maastricht category III). The panel recommends following the withdrawal of life-sustaining therapies protocol usually applied in each unit, which may or may not include immediate extubation. The organ retrieval process should be halted if death does not occur within 3 h of life-support discontinuation. Circulatory arrest is defined as loss of pulsatile arterial pressure and should be followed by a 5-min no-touch observation period. Death is declared based on a list of clinical criteria assessed by two senior physicians. The no-flow time should be no longer than 30, 45, and 90 min for the liver, kidneys, and lungs, respectively. At present, the panel does not recommend pediatric heart donation after death by circulatory arrest. The mean arterial pressure cutoff that defines the start of the functional warm ischemia (FWI) phase is 45 mmHg in patients older than 5 years and/or weighing more than 20 kg. The panel recommends normothermic regional perfusion in these patients. The FWI phase should not exceed 30 and 45 min for retrieving the pancreas and liver, respectively. There is no time limit to the FWI phase for the lungs and kidneys. The panel recommends routine sharing of experience with Maastricht-III donation among all healthcare institutions involved in order to ensure optimal outcome assessment and continuous discussion on the potential difficulties, notably those related to the management of normothermic regional perfusion in small children.

Organ donation by Maastricht-III pediatric patients: Recommendations of the Groupe Francophone de Réanimation et Urgences Pédiatriques (GFRUP) and Association des Anesthésistes Réanimateurs Pédiatriques d'Expression Française (ADARPEF) Part I: Ethical considerations and family care.

The French Transplant Health Authority (Agence de la Biomédecine) has broadened its organ- and tissue-donation criteria to include pediatric patients whose death is defined by circulatory criteria and after the planned withdrawal of life-sustaining therapies (WLST) (Maastricht category III). A panel of pediatric experts convened to translate data in the international literature into recommendations for organ and tissue donation in this patient subgroup. The panel estimated that, among children aged 5 years or over with severe irreversible neurological injury (due to primary neurological injury or post-anoxic brain injury) and no progression to brain death, the number of potential donors, although small, deserves attention. The experts emphasized the importance of adhering strictly to the collegial procedure for deciding to withdraw life support. Once this decision is made, the available data should be used to evaluate whether the patient might be a potential donor, before suggesting organ donation to the parents. This suggestion should be reserved for parents who have unequivocally manifested their acceptance of WLST. The discussion with the parents should include both the pediatric intensive care unit (PICU) team under the responsibility of a senior physician and the hospital organ- and tissue-procurement team. All recommendations about family care during the end of life of a child in the PICU must be followed. The course and potential challenges of organ donation in Maastricht-III pediatric patients must be anticipated. The panel of experts recommended strict compliance with French recommendations (by the Groupe Francophone de Réanimation et Urgences Pédiatriques) about WLST and providing deep and continuous sedation until circulatory arrest. The experts identified the PICU as the best place to implement life-support discontinuation and emphasized the importance of returning the body to the PICU after organ donation. French law prohibits the transfer of these patients from one hospital to another. A description of the expert-panel recommendations regarding the organization and techniques appropriate for children who die after controlled circulatory arrest (Maastricht III) is published simultaneously in the current issue of this journal..

The crucial choice of reference genes: identification of miR-191-5p for normalization of miRNAs expression in bone marrow mesenchymal stromal cell and HS27a/HS5 cell lines.

Bone marrow mesenchymal stromal cells (BM-MSCs) have a critical role in tissue regeneration and in the hematopoietic niche due to their differentiation and self-renewal capacities. These mechanisms are finely tuned partly by small non-coding microRNA implicated in post-transcriptional regulation. The easiest way to quantify them is RT-qPCR followed by normalization on validated reference genes (RGs). This study identified appropriate RG for normalization of miRNA expression in BM-MSCs and HS27a and HS5 cell lines in various conditions including normoxia, hypoxia, co-culture, as model for the hematopoietic niche and after induced differentiation as model for regenerative medicine. Six candidates, namely miR-16-5p, miR-34b-3p, miR-103a-3p, miR-191-5p, let-7a-5p and RNU6A were selected and their expression verified by RT-qPCR. Next, a ranking on stability of the RG candidates were performed with two algorithms geNorm and RefFinder and the optimal number of RGs needed to normalize was determined. Our results indicate miR-191-5p as the most stable miRNA in all conditions but also that RNU6a, usually used as RG is the less stable gene. This study demonstrates the interest of rigorously evaluating candidate miRNAs as reference genes and the importance of the normalization process to study the expression of miRNAs in BM-MSCs or derived cell lines.

Abalakov: Posterior mandibular anchorage technique for orthodontic treatment.

INTRODUCTION: Abalakov is a mountaineering technique consisting in drilling two holes that intersect to form a V in order to thread a rope through to make an anchorage point. This technique can be applied to orthodontic treatment and constitutes a posterior mandibular orthodontic anchorage. TECHNICAL NOTE: The technique can be performed on its own during local anaesthesia or during surgery for the extraction of wisdom teeth. The posterior anchorage point is located in the ramus of the mandible. It consists in drilling two holes that intersect to form a V and then threading a steel wire through to make an anchorage point. DISCUSSION: This technique is simple, inexpensive, fast and non-invasive, providing an anchorage system with immediate loading and which is not dependent on the quality of the bone.

Genomic organization and transcription of satellite DNA in the ant Aphaenogaster subterranea (Hymenoptera, Formicidae).

A satellite DNA family (APSU) was isolated and characterized in the ant Aphaenogaster subterranea. This satellite DNA is organized in tandem repeats of 162 bp and is relatively AT rich (51.9%). Sequence analysis showed a high level of homogeneity between monomers. Loss of satellite DNA has been detected in queens in relation to workers, because the amount of satellite DNA in queens is about 25% of the amount found in workers. Restriction analysis of the total DNA with methylation-sensitive enzymes suggests that this DNA is not methylated. Analysis of the electrophoretic mobility of satellite DNA on non-denaturing polyacrylamide showed that this satellite DNA is only very lightly curved. Their possible transcription was analyzed using reverse transcription and polymerase chain reaction (RT-PCR). The satellite DNA is transcribed on the two DNA strands at the same level in worker and queen pupae, as well as in worker adults.

Tissular distributions and kinetics of two renal 99mTc-radiopharmaceuticals in rat.

A digital radioimager (RI), conventional radioautography (RA), and tracks microradioautography (MRA) were used to assess the biodistributions and kinetics of 99mTc-dimercaptosuccinic (99mTc-DMSA) and 99mTc-mercaptoacetyltriglycine (99mTc-MAG3) in rat at both macroscopic and microscopic levels. Three groups of male Wistar rats were studied. Using gamma-counting, kidney, liver, spleen and blood kinetics of both tracers were assessed in the three groups. Using RA and RI, renal slices were analyzed in group 1 the animals being sacrified from 2 to 60 min after injection of 99mTc-MAG3, and in group 2 the animals being sacrificed from 0.5 to 24 hr after injection of 99mTc-DMSA. Using MRA, renal slices were analyzed for 99mTc-DMSA (group 3). RA films and RI images displayed the variation with time of the cortical and medullary uptakes of the tracers. No regional heterogeneity within the different structures could be seen neither with RA films nor with MRA. The remaining activity in the blood 24 hr after injection of 99mTc-DMSA was evaluated. The tissular distributions of both tracer being homogenous, mean values of cortical uptake seems to be acceptable for dosimetric studies. Our results incite to use of 99mTc-MAG3 instead of 99mTc-DMSA when both tracers may be indicated.

Scintigraphic screening for renal damage in siblings of children with symptomatic primary vesico-ureteric reflux.

OBJECTIVE: To define prospectively the incidence of renal parenchymal lesions in the siblings of patients treated at one institution for primary vesico-ureteric reflux (VUR). PATIENTS AND METHODS: From January 1997 to October 1998, a prospective study including renal scintigraphy (using dimercaptosuccinic acid, DMSA) and a radionuclide cystogram was proposed systematically to the asymptomatic siblings of children treated for primary VUR. The radionuclide cystograms were interpreted as showing the presence or absence of VUR and the DMSA scan as symmetrical or asymmetrical differential function, with or with no renal defect. RESULTS: Fifty-five families gave informed consent, of whom 46 completed the study (eight refused secondarily and one was omitted by exclusion criteria), representing 46 symptomatic patients and 65 siblings. There were 17 siblings with VUR (26%) including two of 13 infants and 15 of 52 children aged > 18 months. One radionuclide cystogram failed. Of the 17 refluxing siblings, four had a history of symptomatic urinary tract infection; 62 of the 65 siblings had a DMSA scan, of which 56 were normal and six (10%) showed abnormalities (five asymmetrical differential function and one parenchymal defect). Only one of these six patients had VUR at the time of the evaluation and only one had a small kidney detected by ultrasonography on one side (and no VUR). There were no adverse effects associated with screening. CONCLUSION: This study confirms a significant overall incidence of VUR (26%) in the asymptomatic siblings of patients treated for primary VUR. From the results of the DMSA scan (only one sibling had a parenchymal defect), the systematic screening of asymptomatic siblings does not appear to be beneficial.

Contribution of single-photon emission computed tomography in the diagnosis and follow-up of CNS toxicity of a cytarabine-containing regimen in pediatric leukemia.

PURPOSE: Cytarabine (ara-C) is one of the most effective chemotherapeutic agents in patients with acute leukemia (AL), with a clear dose effect. Use of high-dose ara-C is hampered, however, by a noticeable toxicity, particularly to the CNS. We investigated the usefulness of CNS perfusion imaging with technetium-99m ((99m)Tc)-hexamethyl-propylene-amine oxime (HMPAO) single-photon emission computed tomography (SPECT) concurrent to magnetic resonance imaging (MRI) to specifically assess the effects of standard- and high-dose ara-C in children with AL. PATIENTS AND METHODS: Twenty-six perfusion studies using (99m)Tc-HMPAO SPECT were performed in 12 children (age range, 4 to 15 years) with AL after induction therapy, which consisted of a standard-dose ara-C, immediately after consolidation with high-dose ara-C, and later during follow-up (range, 6 to 44 months). The chemotherapy-related adverse events were monitored and correlated to SPECT and MRI. RESULTS: After the induction phase, all children were neurologically normal on MRI. On SPECT imaging, four children displayed a slightly heterogeneous perfusion. After high-dose ara-C (4 to 36 g/m(2)), five children had regressive neurologic signs of potential toxic origin. Of these five children, only one had an abnormal MRI scan, whereas all patients showed evidence of diffuse cerebral and/or cerebellar heterogeneous perfusion on SPECT. The seven other patients without any neurologic symptoms had normal MRI scans; SPECT was normal for three patients and abnormal for four patients. On follow-up, for four children who had presented with clinical neurologic toxicity, SPECT improved in three patients and remained unchanged in one patients. In two of these four children, delayed abnormalities (T2 white matter hypersignal and cerebellar atrophy) appeared on MRI scans. CONCLUSION: In our series, diffuse heterogeneous brain hypoperfusion is often the sole early objective imaging feature identified by SPECT of high-dose ara-C neurotoxicity, where MRI still demonstrates normal pictures.

Satellite DNA transcription in Diadromus pulchellus (Hymenoptera).

Previous studies have shown that the satellite DNAs in Hymenoptera account for 1-25% of the genome. They mainly correspond to a single family, or to several subfamilies having the same evolutionary origin. We have now showed that the satellite DNAs in the genomes of the hymenopterans Diadromus pulchellus, Diadromus collaris, Eupelmus vuilletti and Eupelmus orientalis are transcribed in both males and females. Satellite DNA transcripts could only be extracted with NP40/Urea, indicating that they are strongly associated with proteins. The satellite DNA in D. pulchellus was transcribed on the two DNA strands. The satellite DNA transcripts were single-stranded and not polyadenylated in vivo. The transcripts were found in embryos, larvae and imagos stages. The transcripts detected included one major transcript (1.9 kb) and several discrete smaller transcripts. The in vivo synthesis of these satellite DNA transcripts was explored by identifying their putative initiation sites.

Free serosal cells originating from the embryo of the wasp Diadromus pulchellus in the pupal body of parasitized leek-moth, Acrolepiosis assectella. Are these cells teratocyte-like?

In braconid species, teratocytes are derived from a serosal cell membrane which envelops the developing parasitoid embryo. On hatching, this membrane dissociates into individual cells, the teratocytes, which then circulate in the haemolymph of the host. We describe herein such a membrane, surrounding the embryo in eggs of the ichneumonid parasitoid wasp, Diadromus pulchellus. This membrane consisted of a single sheet of tightly packed cells with large 12+/-1.4 &mgr;m nuclei. These cells were released after hatching in vitro and cells of the same size were detected in vivo, in the vicinity of the D. pulchellus embryo. The number of nuclei detected suggests that the serosal membrane consists of about 450+/-150 cells. These cells did not grow after hatching of the parasitoid egg in the parasitized host, Acrolepiosis assectella, during the development of the parasitoid wasp larva. Southern blot experiments, using D. pulchellus satellite DNA or the ribosomal genes as probes, showed that free-living floating cells of wasp origin were present in the body of the parasitized host. This is the first time that free-floating teratocyte-like cells have been described in species of the Ichneumonidae.

Quantitative autoradiography using a radioimager based on a multiwire proportional chamber.

Determination of the biodistribution of radiopharmaceuticals is an important issue for the evaluation of their performance in diagnosis and therapy. In this study, we evaluated a digital radioimager (RI) based on a multiwire proportional chamber for quantitative autoradiography (AR). The RI allows direct detection of electronic emissions of gamma emitters. Its qualitative and quantitative performances were tested on 99mTc and (111)In labelled sections and compared with conventional film AR. Linearity of count rate versus activity was verified over a 104 range of activity. As compared with film AR, a substantial improvement of the detection limit was obtained even for acquisition periods up to 20 times less than film exposure times. We provided the basis for quantitative analysis with tissue equivalent paste standards: the 99mTc and (111)In RI counting efficiencies were respectively 1.19% and 2.35%. We illustrated the respective values of RI and film AR in two rat studies: 99mTc-DMSA in kidney and dual-isotope 99mTc-MIBI and (111)In-antimyosin in heart. Calculated activity concentrations on sections of rat organs confirmed good correlation to gamma counting (deviation less than 12%). We suggest RI as a convenient technique for fast localization of single or dual-isotope tracers and determination of activity distribution.

[Urology and nuclear medicine in children. Experience from a series of 1200 renal scintigraphies]. / Médecine nucléaire en urologie pédiatrique. A propos d'une série de 1,200 scintigraphies rénales.

During the last 4 years, we have performed 1200 renal scintigraphies in children under the age of 6 years: 57% of dynamic renal scintigraphies using MAG3 for antenatally diagnosed uropathies (mainly pelvic dilatations and megaureters), 36% of static renal scintigraphies using DMSA for renal sequelae of pyelonephritis with or without vesicoureteric reflux, and 6% of direct isotope cystography for follow-up of vesicoureteric reflux. Renal scintigraphy, which provides low radiation hazards (1 mSv), is now a major imaging modality for paediatric urinary tract disease.

[First-line vesico-psoas hitch ureteroneocystostomy for primary obstructive megaureter]. / Méga-uretère primitif obstructif: réimplantation sur vessie psoïque de première intention.

Between 1989 and 1997, 23 children with primary obstructive megaureter underwent surgical repair, routinely using a vesico-psoas hitch ureteroneocystostomy. Ten of them had a antenatal diagnosis. Mean postoperative follow-up was 3 years. No postoperative obstruction and no vesicoureteric reflux occurred. Assessment of renal function on the operated side showed improvement in 7 cases and stabilization in all other cases. No bladder dysfunction was observed. Vesicopsoas hitch is an advantageous procedure in this indication. It is almost always unnecessary to perform ureteral tailoring and its specific complications can be avoided. Psoas hitch is easily performed in infants and young children and this technique is particularly appropriate in the subgroup of patients with antenatal diagnosis.

[Ureteropyelostomy for obstructed duplicated ureter an easy and reliable operation in infants]. / La place de l'anastomose urétéro-pyélique dans le traitement des duplications urétérales pathogènes.

AIM: Ureteropyelostomy is one of the surgical options to treat obstructed duplicated ureter. We retrospectively analysed the results of this procedure in a group of young children with prenatal diagnosis. MATERIAL AND METHODS: Between 1992 and 1996, 11 children underwent ureteropyelostomy for obstructed functioning duplicated ureter using the following procedures. 7 cases were revealed by prenatal ultrasound and 4 by acute pyelonephritis; an ureterocele was identified in 7 cases. The mean age at surgery was 6 months for the whole group (3-46 weeks), while in the subgroup of prenatal diagnosis, it was 8 weeks (3-9 weeks). The follow-up ranged from 6 months to 4 years. RESULTS: There were no postoperative complications. None of the anastomoses needed revision, and the conserved segments maintained their functioning appearance, on follow-up. 3 children needed a subsequent ureterovesical reimplantation for urinary tract infections due to reflux in the lower segment ureter. DISCUSSION: Ureteropyelostomy is of great interest for the management of the obstructed dilated upper pole ureter revealed by prenatal diagnosis. This specific group of children needs early management to avoid septic complications and ureteropyelostomy is an easy and reliable method at this young age.

[Nuclear medicine in children]. / Médecine nucléaire chez l'enfant.

Pediatric nuclear medicine (PNM) is now well established as a major imaging modality in children. Recent technical advances in gamma cameras and associated data processing, together with the development of new tracers has modified routine practice in PNM. In order to better ascertain how PNM is routinely used in France, all nuclear medicine departments in Paris and its suburbs answered a detailed questionnaire. The results show three main indications for PNM: exploration of the skeleton (33.3%), the urinary tract (23.5%) and the lung (22.5%).

Complementarity of lung scintigraphy and D-dimer test in pulmonary embolism.

D-dimer assay (DDA), measuring fibrin degradation products, was compared with lung scintigraphy (LS) in a prospective unselected series of 83 consecutive patients referred owing to suspicion of pulmonary embolism (PE). This patient series was also used to compare several methods of performing and interpreting LS images. The final diagnosis was established independently by a separate panel with all available information except for the result of DDA. D-dimer was determined by ELISA (threshold value 500 ng/ml). LS, including perfusion (.Q) and pseudo-ventilation (Technegas) (.V), was classified according to PIOPED, (1) immediately by the physician on duty, and (2) retrospectively by a blinded panel. A positive (19) or negative (61) diagnosis of PE was achieved in 80 patients, the prevalence of PE being 24%. Only one false-negative was noted on DDA (sensitivity=95%) but there were 42 false-positives (specificity=31%), resulting in a positive predictive value of 30% and a negative predictive value of 95%. Emergency and retrospective interpretations of LS were close (kappa=0.4). In a minority of patients, PE may be excluded with reasonable certainty if DDA is normal, resulting in a significant saving in terms of time and money.

Auxiliary liver transplantation in patients with fulminant hepatic failure: hepatobiliary scintigraphic follow-up.

Auxiliary liver transplantation (ALT), retaining in place the liver of the recipient, has been proposed as an alternative to liver replacement in patients with fulminant hepatic failure (FHF). Hepatobiliary scintigraphy (HS) has proved a unique tool for the separate assessment of graft and native liver function. Forty-eight HS scans were performed, following the injection of technetium-99m trimethyl-bromo-imino-diacetic acid, in six patients who underwent ALT for FHF. Quantitative parameters were derived from the time-activity curves of both the graft and the native liver. The function of the graft remained normal as long as the patients remained under immunosuppressive therapy (IST). The function of the native liver was almost completely absent in the 1st month in five patients, but it improved gradually in four of them. IST was then decreased in four patients and finally withdrawn in three. Spontaneous graft atrophy occurred in two patients and the graft was removed in two. All of the patients in whom IST was reduced had a normal global hepatic function and selective uptake (RU) >30% at that time. In ALT patients with FHF, HS can distinguish non-invasively the functional performance of both the donor and the recipient liver and its evolution with time.

Transcription of four satellite DNA subfamilies in Diprion pini (Hymenoptera, Symphyta, Diprionidae).

Four satellite DNA subfamilies Ps, Pv, Pv65 and Ec, resulting from the evolution of a common ancestral motif, were isolated and characterized in the genomic DNA of Diprion pini, a phytophagous of Pinus sylvestris. Consensus sequences were 148-312 bp long. Sequence analyses revealed that these satellite subfamilies have evolved from a 45-bp ancestral motif. The amounts of each satellite in the genome (0 - 10%) and the accessibility of the DNA to restriction enzymes were sex dependent. The migration of each monomer in polyacrylamide gels and the electrophoretic migration of d(AT) n > or = 3 residues showed that all four satellite subfamilies are curved. Their transcription was analyzed using reverse transcription and the polymerase chain reaction. Three satellite DNA subfamilies were transcribed on both strands, and in both sexes. However, the female satellite DNAs seem to be more actively transcribed than those of males, indicating that transcription is not constitutive. The lack of any significant open reading frame in satellite monomers indicates that the RNA may function as structural or catalytic RNA rather than encoding protein.