Diversity and composition of macroinvertebrate communities in a rare inland salt marsh.

Inland salt marshes are rare habitats in the Great Lakes region of North America, formed on salt deposits from the Silurian period. These patchy habitats are abiotically stressful for the freshwater invertebrates that live there, and provide an opportunity to study the relationship between stress and diversity. We used morphological and COI metabarcoding data to assess changes in diversity and composition across both space (a transect from the salt seep to an adjacent freshwater area) and time (three sampling seasons). Richness was significantly lower at the seep site with both datatypes, while metabarcoding data additionally showed reduced richness at the freshwater transect end, consistent with a pattern where intermediate levels of stress show higher diversity. We found complementary, rather than redundant, patterns of community composition using the two datatypes: not all taxa were equally sequenced with the metabarcoding protocol. We identified taxa that are abundant at the salt seep of the marsh, including biting midges (Culicoides) and ostracods (Heterocypris). We conclude that (as found in other studies) molecular and morphological work should be used in tandem to identify the biodiversity in this rare habitat. Additionally, salinity may be a driver of community membership in this system, though further ecological research is needed to rule out alternate hypotheses.

Basal Ti level in the human placenta and meconium and evidence of a materno-foetal transfer of food-grade TiO2 nanoparticles in an ex vivo placental perfusion model.

BACKGROUND: Titanium dioxide (TiO2) is broadly used in common consumer goods, including as a food additive (E171 in Europe) for colouring and opacifying properties. The E171 additive contains TiO2 nanoparticles (NPs), part of them being absorbed in the intestine and accumulated in several systemic organs. Exposure to TiO2-NPs in rodents during pregnancy resulted in alteration of placental functions and a materno-foetal transfer of NPs, both with toxic effects on the foetus. However, no human data are available for pregnant women exposed to food-grade TiO2-NPs and their potential transfer to the foetus. In this study, human placentae collected at term from normal pregnancies and meconium (the first stool of newborns) from unpaired mothers/children were analysed using inductively coupled plasma mass spectrometry (ICP-MS) and scanning transmission electron microscopy (STEM) coupled to energy-dispersive X-ray (EDX) spectroscopy for their titanium (Ti) contents and for analysis of TiO2 particle deposition, respectively. Using an ex vivo placenta perfusion model, we also assessed the transplacental passage of food-grade TiO2 particles. RESULTS: By ICP-MS analysis, we evidenced the presence of Ti in all placentae (basal level ranging from 0.01 to 0.48 mg/kg of tissue) and in 50% of the meconium samples (0.02-1.50 mg/kg), suggesting a materno-foetal passage of Ti. STEM-EDX observation of the placental tissues confirmed the presence of TiO2-NPs in addition to iron (Fe), tin (Sn), aluminium (Al) and silicon (Si) as mixed or isolated particle deposits. TiO2 particles, as well as Si, Al, Fe and zinc (Zn) particles were also recovered in the meconium. In placenta perfusion experiments, confocal imaging and SEM-EDX analysis of foetal exudate confirmed a low transfer of food-grade TiO2 particles to the foetal side, which was barely quantifiable by ICP-MS. Diameter measurements showed that 70 to 100% of the TiO2 particles recovered in the foetal exudate were nanosized. CONCLUSIONS: Altogether, these results show a materno-foetal transfer of TiO2 particles during pregnancy, with food-grade TiO2 as a potential source for foetal exposure to NPs. These data emphasize the need for risk assessment of chronic exposure to TiO2-NPs during pregnancy.

How do parents perceive adverse drug events of their children's anticonvulsant medication?

BACKGROUND: The main source of knowledge on adverse drug events (ADE) are physicians' reports in controlled clinical trials. In contrast, little is known about the parents' perception of ADE of anticonvulsants their children receive. METHODS: After approval by the local ethics committee, we performed a survey in a neuropediatric outpatient clinic of a university hospital. Based on a structured questionnaire, we interviewed parents of children with current anticonvulsant treatment regarding (i) their fears about potential ADE, (ii) experienced ADE according to parents, and (iii) implications of ADE on the child's life. RESULTS: Parents of 150 patients took part in the interview. (i) 95 (63.3%) parents expressed fears concerning ADE, mostly liver injury/liver failure (33 [22%]). (ii) 129 (86%) parents reported experienced ADE, mostly sedation (65 [43.3%]) and abnormal behavior (54 [36%]). (iii) Parents reported substantial implications of ADE on the child's daily life for 84 (56%) children, and 63 (42%) parents expressed a negative impact on the child's development. CONCLUSION: We recognized a great discrepancy between those ADE that were feared and those that were experienced. Parents feared life-threatening ADE and experienced less severe ADE that nevertheless have a negative impact on the child's daily life.

Impact of prenatal and postnatal exposure to the pesticide chlorpyrifos on the contraction of rat ileal muscle strips: involvement of an inducible nitric oxide synthase-dependent pathway.

BACKGROUND: Prenatal/postnatal exposure to insecticides has been linked to developmental disorders in adulthood. Chlorpyrifos (CPF) is a widely used organophosphorus acetylcholinesterase (AChE)-inhibiting insecticide. The present study established whether prenatal and postnatal exposure to CPF is associated with intestinal motor dysfunction in adult rats. METHODS: Three groups of pregnant rats were exposed to either CPF (1 or 5 mg/kg/day; the CPF1 and CPF5 groups) or vehicle (the control group) by gavage from gestational day 1 until weaning. At weaning, the pups were separated from their dams and individually gavaged (with the same dose) until postnatal day 60. We then measured in vivo intestinal transit and the in vitro contractile responses of ileal smooth muscle strips to electrical field stimulation. Expression of inducible nitric oxide synthase (iNOS) in the ileum was determined using qRT-PCR and immunoblots. Morphometry and AChE assays were also performed. KEY RESULTS: At adulthood, the mean body mass was lower in the CPF1 and CPF5 groups than in controls. CPF5 exposure was associated with weaker in vitro contraction of ileal muscle strips, which was reversed by adding the NOS inhibitor (L-NAME). There was no significant intergroup difference in the mean in vivo transit time. Exposure to CPF was associated with greater iNOS expression, lower AChE activity and reduced circular and longitudinal smooth muscle thickness. CONCLUSIONS & INFERENCES: Prenatal and postnatal exposure to CPF in the rat is associated with weaker contraction of ileal longitudinal smooth muscle via a nitrergic mechanism with increased iNOS expression.

[Evaluation of the measurement of hemoglobin by the Hemocue System® in the preterm neonate less than 28 days old]. / Évaluation de la mesure du taux d'hémoglobine par Hémocue® chez le nouveau-né prématuré de moins de 28 jours de vie.

BACKGROUND: Hemoglobin (Hb) measurement is essential for the monitoring of anemia in preterm neonates to assess if any bleeding (pulmonary, cerebral, digestive) is present. EDTA samples require 500 µL vs. 10 µL for the Hemocue(®) system. This system has been evaluated and validated in adults and children but not in preterm neonates with fetal hemoglobin. The aim of the study was to compare Hb measurement with the Hemocue(®) system vs. the EDTA laboratory system on fetal Hb in preterm neonates. MATERIALS AND METHODS: This was a prospective study conducted in the preterm intensive care unit in the Amiens Hospital. Preterm neonates, before 28 days of life, requiring EDTA (Hb) measurement were included. Two Hemocues(®) were performed at the same time. Postnatal age (correlated to the fetal hemoglobin level decrease), blood sample site, and other factors that could influence the Hb result were evaluated. RESULTS: Seventy-six EDTA and 152 Hemocue(®) samples from 38 preterm neonates were included. The term was 28.1±3.7 weeks of gestation, the birth weight was 1215.5±657 g. We found a good correlation between the Hemocue(®) and EDTA samples (Hemocue(®)=EDTA*0.94-0.4; R(2)=0.63; P<0.001). The influence of confounding factors was insignificant. CONCLUSION: The use of the Hemocue(®) system showed a good correlation with the EDTA measurement of fetal Hb, with a moderate bias (-0.2±1.5 g/dL), which remained stable for the first 28 days of life.

A mean body temperature of 37°C for incubated preterm infants is associated with lower energy costs in the first 11 days of life.

AIM: This randomised trial compared the energy costs of providing incubated preterm infants born before 32 weeks of gestation with homeothermia using either air temperature control (ATC) or skin servocontrol (SSC). METHODS: We studied 38 incubated preterm infants for the first 11 days of life, calculating the frequency of hypothermia (<36.0°C), hyperthermia (>37.5°C) and thermal challenge, together with energy costs, based on a change in incubator air temperature of 2°C above or below thermoneutrality. RESULTS: The daily mean incubator air temperature was higher in ATC than SSC (p < 0.05) for the first 6 days, and the mean body temperature was higher in ATC (37.0 ± 0.03°C) than SSC (36.8 ± 0.02; p < 0.01) over the whole study period. The frequency of moderate hyperthermia was higher in ATC (p < 0.001), whereas warm and cold thermal challenges were higher in SSC (p < 0.001). The two groups did not differ in terms of energy costs. The time to recover birthweight was shorter in ATC (p < 0.05). CONCLUSION: In incubators using ATC, a body temperature of 37°C was associated with lower energy costs and greater weight gain at 11 days of life for preterm infants. Future studies should test SSC shielded abdominal skin temperature set to 37°C.

Usefulness of (1,3) ß-D-glucan detection in bronchoalveolar lavage samples in Pneumocystis pneumonia and Pneumocystis pulmonary colonization.

OBJECTIVE OF THE STUDY: Recent data demonstrate the usefulness of (1,3) ß-d-glucan (BG) detection in serum samples to distinguish patients developing Pneumocystis pneumonia and patients who are colonized by the fungus. In contrast, data of BG detection in bronchoalveolar lavage (BAL) samples from these patient populations are still rare. PATIENTS: In this context, we determined BG levels in BAL samples from 11 Pneumocystis pneumonia (PCP) patients, 10 colonized patients, and 24 Pneumocystis-uninfected patients. MATERIALS AND METHODS: BG levels were determined on each BAL sample using the Fungitell(®) kit (Associates of Cape Cod, Inc., Cape Cod, MA, USA) according to the manufacturer's instructions applied to serum sample examination. RESULTS: The BG levels in BAL samples from the PCP patient group (mean value 20 588 pg/mL) were significantly higher than those in the colonized patient group (mean value 105 pg/mL) (P=0.0001, Mann-Whitney test) and than those in the Pneumocystis-uninfected patient group (mean value 74 pg/mL) (P<0.0001, Mann-Whitney test). The BG levels in BAL samples from the colonized patient group did not differ significantly from those in the Pneumocystis-uninfected patients group (P=0.21). CONCLUSION: The results suggest that measurements of BAL BG levels may facilitate the differential diagnosis of PCP and pulmonary colonization with Pneumocystis.

[Does insulin pump therapy improve quality of life and satisfaction in children and adolescents with type 1 diabetes?]. / La pompe à insuline améliore-t-elle la qualité de vie et la satisfaction des enfants et adolescents diabétiques de type 1 ?

Insulin pumps are booming in pediatric diabetology. The objective of this study was to assess changes for children and adolescents with type 1 diabetes using a pump in terms of quality of life (QOL), satisfaction, and glycosylated hemoglobin. A retrospective self-evaluation questionnaire was distributed to 41 patients. It focused on general QOL, diabetes-specific QOL supplemented by specific questions on the pump, and satisfaction. Clinical and biological parameters (glycated hemoglobin: HbA1c) were compared before and after pump use. The score for QOL with the pump was positive, more so if started early after diagnosis of diabetes (P=0.03) and with children under the age of 8 years (P<0.02). These positive results are mainly related to the characteristics of the pump, "insulin management" and "injections," as well as "diabetes management," "behavior," "school," "family life," "daily life," and "physical activities." On the other hand, the improvement was not significant for the item "life in society, friends and family." A decrease in the number of injections and the flexibility of meals were the most positive points. HbA1c improved as soon as the pump was indicated before its use was begun (P=0.005) and remained constant for 4 years (P≤0.05). Forgotten injections, comments on diabetes, and technical problems appeared to be exceptional. The pump changed the patient's body image because of ambivalent feelings between being normal (greater freedom) and different (visibility and a reminder of the disease). The benefits in terms of QOL and glycemic control with the pump cannot be dissociated and can only be considered accompanied by paramedical and medical assistance. Improving QOL over the short and long term by reducing the risk of further complications is the daily challenge of families and diabetologists.

Perinatal nicotine/smoking exposure and carotid chemoreceptors during development.

Tobacco smoking is still a common habit during pregnancy and is the most important preventable cause of many adverse perinatal outcomes. Prenatal smoking exposure can produce direct actions of nicotine in the fetus with the disruption of body and brain development, and actions on the maternal-fetal unit by causing repeated episodes of hypoxia and exposure to many toxic smoke products (such as carbon monoxide). Specifically, nicotine through binding to nicotinic acetylcholine receptors have ubiquitous effects and can affect carotid chemoreception development through structural, functional and neuroregulatory alterations of the neural circuits involved in the chemoafferent pathway, as well as by interfering with the postnatal resetting of the carotid bodies. Reduced carotid body chemosensitivity and tonic activity have thus been reported by the majority of the human and animal studies. This review focuses on the effects of perinatal exposure to tobacco smoke and nicotine on carotid chemoreceptor function during the developmental period. A description of the effects of smoking and nicotine on the control of breathing related to carotid body activity, and of the possible physiopathological mechanisms at the origin of these disturbances is presented.

Impact of nursing care on temperature environment in preterm newborns nursed in closed convective incubators.

AIMS: Very-low-birth-weight (VLBW) neonates require regular nursing procedures with frequent opening of the incubator resulting in a decrease in incubator air temperature. This study was designed to assess changes in the thermal status of VLBW neonates according to the type of nursing care and incubator openings. METHODS: Thirty-one VLBW neonates (mean gestational age: 28.7 ± 0.3 weeks of gestation) were included. Over a 10-day period, each opening of the incubator was recorded together with details about caregiving. Body temperature was recorded continuously, and door opening and closing events were recorded by a video camera. RESULTS: This study analysed 1,798 caregiving procedures with mean durations ranging from 6.2 ± 2.1 to 88.5 ± 33.4 min. Abdominal skin temperature decreased by up to 1.08°C/h for procedures such as tracheal intubation (p < 0.01). The temperature decrease was strongly correlated with the type of procedure (p < 0.01), incubator opening (p < 0.01) and procedure duration (p < 0.01). The procedure duration accounted for only 10% of the abdominal skin temperature change (p < 0.01). CONCLUSIONS: For VLBW neonates nursed in skin temperature servo-control incubators, the decrease in abdominal skin temperature during caregiving was correlated with the type of procedure, incubator opening modalities and procedure duration. These parameters should be considered to optimize the thermal management of VLBW neonates.

Spectral and copper binding properties of methanobactin from the facultative methanotroph Methylocystis strain SB2.

Methanobactin (mb) is the first characterized example of a chalkophore, a class of copper-binding chromopeptides similar to iron-binding siderophores. Structural, redox, themodynamic, and spectral studies on chalkophores have focused almost exclusively on the mb from Methylosinus trichosporium OB3b (mb-OB3b). The structural characterization of a second mb from Methylocystis strain SB2 (mb-SB2) provides a means to examine the core structural features and metal binding properties of this group of chromopeptides. With the exception of the 5-membered rings (either oxazolone or imidazolone), enethiol groups, and the N-terminus oxo group, the structure of mb-SB2 differs markedly from mb-OB3b. In particular the amino acids commonly associated with metal coordination and redox activity found in mb-OB3b, Cys, Met, and Try, are replaced by Ala or are missing in mb-SB2. In this report the spectral and thermodynamic properties of mb-SB2 are presented and compared to mb-OB3b. The results demonstrate that the spectral and basic copper binding properties of both methanobactins are similar and the unique copper binding capacity of both methanobactins lies primarily in the pair of five-membered rings and associated enethiol groups. The remaining portions of the methanobactin appear to provide the scaffolding that brings together of the two ring systems to produce the tetrahedral binding site for copper binding.

Mast cells mediate Pseudomonas aeruginosa lipopolysaccharide-induced lung inflammation in rat.

Activated mast cells have been demonstrated to play a pivotal role in Pseudomonas aeruginosa lung infections. However, there is no report about the involvement of mast cells in P. aeruginosa lipopolysaccharide (LPS)-induced lung inflammation. This study aimed at evaluating the role of mast cells in P. aeruginosa LPS-induced lung inflammation in rats. Mast cells stabilization was carried out by intraperitoneal injections of cromolyn. Lung inflammation was induced by the intratracheal instillation of P. aeruginosa LPS (5 µg/kg bw) and inflammatory status was evaluated 4 h post-LPS instillation. We found that activated mast cells could constitute a pivotal source of several inflammatory cytokines, including TNF-α, IL-1ß, and IL-6. These cells might regulate polymorphonuclear neutrophil (PMN) recruitment and be implicated in the alteration of alveolar-capillary permeability via the release of TNF-α and IL-1ß. We also detected that activated mast cells could be involved in the alteration of the expression of two epithelial tight junction proteins (claudin-1 and occludin) during the acute phase of inflammation. Our results suggest that activated mast cells might play a critical role in P. aeruginosa LPS-induced lung inflammation. Therefore, mast cell stabilization may be a potential novel approach for the prevention and treatment of P. aeruginosa-induced lung infections.

[Factors related to extubation failure in premature infants less than 32 weeks of gestation]. / Facteurs associés à un échec d'extubation chez le nouveau-né prématuré de moins de 32 semaines d'aménorrhée.

UNLABELLED: Even though ventilatory support for preterm infants has improved over the past few decades, about 30% of intubated preterm infants fail an extubation attempt. There are still no simple standards to determine the optimal extubation time for those infants. OBJECTIVE: To identify factors related to extubation success or failure in preterm infants less than 32 weeks of gestation (WG). POPULATION AND METHODS: Retrospective study including 162 newborns less than 32 WG, requiring mechanical ventilation within the first 24h of life. Successful extubation was defined as no need for reintubation for any reason at least 7 days after the first extubation attempt. RESULTS: Forty-one infants failed extubation (25.3%); 149 infants (92%) had mechanical ventilation in their first 30min of life. These infants had a lower gestational age (p<0.001) and a lower birth weight (p=0.003). They required a higher FiO(2) (p<0.001) and maximum inspiratory pressure (p=0.002). The infants who failed extubation had a lower pH (p=0.001) and hematocrit (p=0.032), and a higher PCO(2) (p=0.003). CONCLUSION: As previously described, a low gestational age is a major factor predicting the extubation failure. However, our data suggests that ventilator settings and blood gas should also be considered when attempting extubation, following written guidelines.

[Heat exchanges and thermoregulation in the neonate]. / Echanges thermiques et thermorégulation chez le nouveau-né.

The newborn's energy expenditure is used in order of priority for: (i) basic metabolism; (ii) body temperature regulation and (iii) body growth. Thermal regulation is an important part of energy expenditure, especially for low birth-weight infants or preterm newborns. The heat exchanges with the environment are greater in the infant than in the adult, explaining the increased risk of body hypo- or hyperthermia. The newborn infant is a homeotherm, but over a long period of time, he cannot maintain the thermal processes. Further developments are expected to improve the infant's thermal environment, with assessment of the various heat exchange mechanisms by conduction, convection, radiation and evaporation. The quantification of the respective parts of these exchanges would improve nursing care through clinical procedures or equipment used to ensure the control of the optimal thermohygrometric conditions in incubators, especially when the likelihood of excessive body cooling is high. The present review focuses on the various body heat exchange mechanisms, the thermoregulation processes of the newborn, and their implications in clinical usage and limitations in the neonatal intensive care unit.

Dramatic improvement in survival after lung transplantation over time: a single center experience.

Lung transplantation (LT) is a recognized procedure for selected patients with end-stage respiratory failure. We performed 123 LT, including 32 single lung, 84 double lung, and 7 heart-lung transplantations in 48 patients with chronic obstructive pulmonary disease (COPD), 13 patients with pulmonary hypertension (PH), 33 with cystic fibrosis (CF), and 29 with interstitial lung disease (ILD) between July 1990 and January 2008. Survival was compared for periods before and after December 2001. The mean age of patients was 44.4 years (range 16-66.5 years); 84 (69%) were men. Before LT, 1 second forced expiratory volume was 28.7% +/- 18.1% and PaCO(2) = 6.3 kPa. Fifty-five patients were on noninvasive ventilation. Cold ischemia time was 320 +/- 91 minutes. Cardiopulmonary bypass (CPB) was used in 77 patients (64%). There were 18 early surgical reinterventions, 8 extracorporeal membrane oxygenations, and 38 bronchial stent insertions among 206 at-risk bronchial sutures. Crude survivals were 69%, 58%, 41%, and 18% at 1, 2, 5, and 10 years, respectively. Comparing before (n = 70 with 15 CF) vs after December 2001 (n = 53 with 17 CF), survivals were 63% vs 78%, 51% vs 71%, and 33% vs 60% at 1, 2, and 5 years, respectively (P = .01) and for CF patients, 52% vs 100%, 52% vs 94%, and 25% vs 94% at 1, 2, and 5 years, respectively (P = .005). There was significant improvement in survival before and after 2001 in 123 LT and particularly among CF patients. Improvement in survival after LT may be related to the sum of numerous changes in our practice since December 2001, including the use of pulmonary rehabilitation pre-LT, extracellular pneumoplegia, statins, macrolides for chronic rejection, monitoring of Epstein-Barr blood load, changes in maintenance immunosuppressants, as well as position movement up the coordinator nurse and learning curve.

Genotyping the HFE gene by melting point analysis with the LightCycler system: Pros and cons.

The assay that combines rapid-cycle PCR with allele-specific fluorescent probe melting profiles performed on the Roche Diagnostics LightCycler is commonly employed for genotyping the HFE gene. We report three illustrative cases of the pros and cons of this method. In two cases, atypical melting curves allows the identification of new DNA substitutions in the HFE gene, whereas, in the third case, a typical melting curve of c.845G>A mutation (C282Y) homozygosity overlooks a nucleotide change and promotes misdiagnosis of HH.

Effect of aprotinin on postoperative blood loss in off-pump coronary artery bypass surgery.

BACKGROUND AND AIM OF THE STUDY: Off-pump coronary artery bypass (OPCAB) enables a reduction in postoperative complications, particularly bleeding and transfusion. Nevertheless, a significant percentage of patients still needs transfusion. The effect of antifibrinolytic therapy on postoperative bleeding as part of OPCAB is still not widely described. The purpose of this study was to investigate the potential benefit of aprotinin in OPCAB. METHODS: We conducted a retrospective comparative study with a historical control group. Consecutive patients undergoing off-pump coronary bypass were divided in two groups: 40 patients were operated without any antifibinolytic drug (group C); 40 patients received aprotinin (group A) during surgery. Patients in group A received a bolus of 2 x 10(6) KIU during 30 minutes, followed by a continuous infusion of 0.5 x 10(6) KIU per hour until the end of surgery. The same protocol was used during the whole study period. RESULTS: Preoperative data of the two groups did not differ except for the number of grafts performed, which was higher in group A. Prothrombin time and activated clotting time increased in both groups after surgery. The use of packed red blood cells or fresh frozen plasma was not significantly different between both groups. Postoperative blood loss was significantly reduced in the aprotinin group (540 mL +/- 320 vs. 770 mL +/- 390, p = 0.006). No increase in postoperative troponin values was found in group A. CONCLUSIONS: Aprotinin significantly reduced postoperative blood loss without reducing the transfusion rate. Aprotinin was not associated with any increase in postoperative complications.

Autosomal dominant hereditary hemochromatosis associated with two novel Ferroportin 1 mutations in Spain.

Hereditary hemochromatosis is a common disorder of iron metabolism most frequently associated with mutations in the HFE gene. Hereditary hemochromatosis may be caused by other genetic mutations including those in the SLC40A1 gene. This report describes the clinical and laboratory findings of two Spanish families with autosomal dominant iron overload associated with previously unrecognized Ferroportin 1 mutations (p.R88T and p.I180T). The phenotype of iron overload in the patients carrying these mutations could correspond to the group of clinical mutations that lose their iron export function.

Mutation analysis of Wilson disease in the Spanish population -- identification of a prevalent substitution and eight novel mutations in the ATP7B gene.

Wilson disease (WD) is a copper metabolism disorder characterized by hepatic and/or neurological damage. More than 200 mutations in the ATP7B gene causing this autosomal recessive defect have been reported. In certain populations, a high prevalence of particular mutations allows rapid screening and diagnosis of the disease. We identified the ATP7B alterations in Spanish patients with WD. Mutations in the ATP7B gene were analysed in a total of 64 individuals from 40 different WD families by PCR amplification, single-strand conformation polymorphism (SSCP) analysis and sequencing. Twenty-one different ATP7B gene mutations were identified, eight of which were novel. 74% of the disease alleles were characterized among the 40 unrelated probands. We identified a prevalent mutation in our population (Met645Arg), present in 55% of this 40 patients. The frequency of the remaining ATP7B alterations was low. In addition, 17 different polymorphic variants were found. There is remarkable allele heterogeneity in WD in the Spanish population. Nevertheless, SSCP screening for the most frequent mutations in our population is feasible and leads to the detection of about 74% of the mutated chromosomes. Molecular diagnosis of WD is very useful in clinical practice to confirm or support clinical suspicion.