Stress during first gestation of ewes impairs memory and learning of male offspring.

This study aimed to investigate the influence of gestational stress induced by lipopolysaccharide (LPS, Escherichia coli) on the physiological changes of ewes, as well as on the subsequent behavioral interaction between ewes and lambs and on the memory and learning of 30-day-old offspring in a T-maze. Thirty-six nulliparous pregnant crossbred Santa Ines ewes with an initial live weight of 45 ± 6 kg, age of 12 ± 2 months, and body condition score between 3 and 3.5 (on a scale of 1 to 5) were divided into two treatments: LPS treatment (E. coli; 0.8 µg.kg-1) and Control (placebo/saline) administered in late pregnancy (day 120). Blood samples were collected before (0 h at 5:00 h) and 1 h, 2 h, 4 h, 8 h, 12 h, 24 h after the administration of LPS or placebo to determine the cortisol release curve. Rectal temperature was measured at the same time points. After birth, male lambs (N = 19) were used to evaluate the maternal-offspring behavioral interaction, weight, and cognitive ability in a T-maze. Blood cortisol and rectal temperature of ewes increased after LPS administration and returned to baseline levels after 24 h. The activities facilitating and stimulating suckling were higher on LPS group (P < 0.05). Lambs whose mothers were challenged with LPS during late pregnancy showed greater learning and memory disabilities including fear behavior and the inability to make decisions at 30 days of age in the T-maze. In sheep, the immunological stress induced by LPS in late pregnancy promotes an inflammatory response characterized by specific rectal temperature and cortisol release profiles, improving maternal care that can increase offspring survival; however, the exposure of sheep fetuses to maternal inflammation causes cognitive impairment in lambs at 30 days of age, which could not be reduced by the behavioral interaction between the mother and offspring.

Evidence of Air Trapping During Ex Vivo Lung Perfusion: A Swine Experimental Lung Imaging and Mechanics Study.

Ex vivo lung perfusion (EVLP) allows the ventilation and perfusion of lungs to evaluate their viability for transplantation. The aim of this study is to compare the mechanical, morphologic and functional properties of lungs during EVLP with values obtained in vivo to guide a safe mechanical ventilation strategy. Lungs from 5 healthy pigs were studied in vivo and during 4 hours of EVLP. Lung compliance, airway resistance, gas exchange, and hemodynamic parameters were collected at positive end-expiratory pressure (PEEP) of 5 cm H2O. Computed tomography was performed at PEEP 0, PEEP 5, and total lung capacity (TLC). Lung pressure-volume (PV) curves were performed from PEEP 0 to TLC. Lung compliance decreased during EVLP (53 ± 5 mL/cm H2O vs 29 ± 7 mL/cm H2O, P < .05), and the PV curve showed a lower inflection point. Gas content (528 ± 118 mL vs 892 ± 402 mL at PEEP 0) and airway resistance (25 ± 5 vs 44 ± 9 cmH2O/L∗s-1, P < .05) were higher during EVLP. Alveolar dead space (5% ± 2% vs 17% ± 6%, P < .05) and intrapulmonary shunt (9% ± 2% vs 28% ± 13%, P < .05) increased ex vivo compared to in vivo, while the partial pressure of oxygen to inspired oxygen fraction ratio (PO2/FiO2) did not differ (468 ± 52 mm Hg vs 536 ± 14 mm Hg). In conclusion, during EVLP lungs show signs of air trapping and bronchoconstriction, resulting in low compliance and increased alveolar dead space. Intrapulmonary shunt is high despite oxygenation levels acceptable for transplantation.

Hospital surge capacity in a tertiary emergency referral centre during the COVID-19 outbreak in Italy.

The first person-to-person transmission of the 2019 novel coronavirus in Italy on 21 February 2020 led to an infection chain that represents one of the largest known COVID-19 outbreaks outside Asia. In northern Italy in particular, we rapidly experienced a critical care crisis due to a shortage of intensive care beds, as we expected according to data reported in China. Based on our experience of managing this surge, we produced this review to support other healthcare services in preparedness and training of hospitals during the current coronavirus outbreak. We had a dedicated task force that identified a response plan, which included: (1) establishment of dedicated, cohorted intensive care units for COVID-19-positive patients; (2) design of appropriate procedures for pre-triage, diagnosis and isolation of suspected and confirmed cases; and (3) training of all staff to work in the dedicated intensive care unit, in personal protective equipment usage and patient management. Hospital multidisciplinary and departmental collaboration was needed to work on all principles of surge capacity, including: space definition; supplies provision; staff recruitment; and ad hoc training. Dedicated protocols were applied where full isolation of spaces, staff and patients was implemented. Opening the unit and the whole hospital emergency process required the multidisciplinary, multi-level involvement of healthcare providers and hospital managers all working towards a common goal: patient care and hospital safety. Hospitals should be prepared to face severe disruptions to their routine and it is very likely that protocols and procedures might require re-discussion and updating on a daily basis.

Development and validation of a multiplex fluorescent microsphere immunoassay assay for detection of porcine cytokines.

Cytokines are cell signalling proteins that mediate a number of different physiological responses. The accurate measurement of cytokine profiles is important for a variety of diagnostic and prognostic scenarios in relation to animal health and welfare. Simultaneous quantification of cytokine profiles in a single sample is now possible using fluorescent microsphere immunoassays (FMIA). We describe the development and validation of a novel multiplex assay using the Bio-Plex® 200 system to quantify cytokines in five different porcine tissues (brain, placenta, synovial tissue and fluid, plasma). The cytokine profiles are both tissue, and research hypothesis, -dependent but include Interleukin-1beta (IL-1ß), Interleukin-4 (IL-4), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10) and Tumor necrosis factor (TNF-α). This methods paper is reported in two parts: the development of a FMIA for porcine tissues and validation of pre-treatment for optimal cytokine recovery in porcine brain, placenta, synovial tissue and plasma. Validation steps are critical in ensuring an assay is suitable for novel sample types. This technique advances traditional ELISAs by: •FMIA provides insight into the profiles of multiple porcine cytokines in certain situations (e.g. disease, parturition).•Use of the Bio-Plex® 200 system to investigate novel sample types, including brain, placenta and synovial tissue.•Multiplexing utilises a fraction of the sample volume compared with multiple ELISAs.

Preliminary Experience With Hypothermic Oxygenated Machine Perfusion in an Italian Liver Transplant Center.

BACKGROUND: Machine perfusion is increasingly utilized in liver transplantation to face the detrimental consequences of the use of extended-criteria donors. Hypothermic oxygenated machine perfusion (HOPE) appears to be more protective relative to static cold storage. Conversely, normothermic machine perfusion (NMP) allows a better graft evaluation. We describe a pilot prospective study on machine perfusion in selected grafts. METHODS: HOPE was executed for all the grafts procured from donors after cardiac death (DCDs) and for livers from donors after brain death (DBDs) requiring prolonged preservation time. NMP was used when a more precise evaluation was needed. Both HOPE and NMP were performed through the portal vein and hepatic artery. RESULTS: From July 2016 to November 2017, we performed 7 HOPE procedures: 5 for DCD and 2 for DBD grafts. Two livers presented with macrovesicular steatosis >30% (1 DCD and 1 DBD). HOPE lasted 240 minutes (180-320 min) with a total ischemia time of 575 minutes (410-810 min). Six grafts were successfully transplanted. One DCD graft required additional evaluation using NMP. The graft was then discarded due to extensive hepatocellular necrosis. In the post-transplant course, acute and chronic renal failure were the main complications affecting 3 and 2 recipients, respectively. In our series, steatosis was the main risk factor for kidney injury. Patient and graft survival rate was 100% and no ischemic cholangiopathies were observed after 270 days (106-582 days). CONCLUSIONS: Our study confirms HOPE safety and efficacy for DCD and DBD grafts. These data are particularly significant for DCD management in the Italian setting where the mandatory 20-minute hands-off interval before death declaration further prolongs warm ischemia time.

Recruitment manoeuvres dislodge mucus towards the distal airways in an experimental model of severe pneumonia.

BACKGROUND: Recruitment manoeuvres generate a transient increase in trans-pulmonary pressure that could open collapsed alveoli. Recruitment manoeuvres might generate very high inspiratory airflows. We evaluated whether recruitment manoeuvres could displace respiratory secretions towards the distal airways and impair gas exchange in a porcine model of bacterial pneumonia. METHODS: We conducted a prospective randomised study in 10 mechanically ventilated pigs. Pneumonia was produced by direct intra-bronchial introduction of Pseudomonas aeruginosa. Four recruitment manoeuvres were applied randomly: extended sigh (ES), maximal recruitment strategy (MRS), sudden increase in driving pressure and PEEP (SI-PEEP), and sustained inflation (SI). Mucus transport was assessed by fluoroscopic tracking of radiopaque disks before and during each recruitment manoeuvre. The effects of each RM on gas exchange were assessed 15 min after the intervention. RESULTS: Before recruitment manoeuvres, mucus always cleared towards the glottis. Conversely, mucus was displaced towards the distal airways in 28.6% ES applications and 50% of all other recruitment manoeuvres (P=0.053). Median mucus velocity was 1.26 mm min-1 [0.48-3.89] before each recruitment manoeuvre, but was reversed (P=0.007) during ES [0.10 mm min-1 [-0.04-1.00]], MRS [0.10 mm min-1 [-0.4-0.48]], SI-PEEP [0.02 mm min-1 [-0.14-0.34]], and SI [0.10 mm min-1 [-0.63-0.75]]. When PaO2 failed to improve after recruitment manoeuvre, mucus was displaced towards the distal airways in 68.7% of the cases, compared with 31.2% recruitment manoeuvres associated with improved PaO2 (odds ratio: 4.76 (95% confidence interval: 1.13-19.97). CONCLUSIONS: Recruitment manoeuvres dislodge mucus distally, irrespective of airflow generated by different recruitment manoeuvres. Further investigation in humans is warranted to corroborate these pre clinical findings, as there may be limited benefits associated with lung recruitment in pneumonia.

An extremely young massive clump forming by gravitational collapse in a primordial galaxy.

When cosmic star formation history reaches a peak (at about redshift z ≈ 2), galaxies vigorously fed by cosmic reservoirs are dominated by gas and contain massive star-forming clumps, which are thought to form by violent gravitational instabilities in highly turbulent gas-rich disks. However, a clump formation event has not yet been observed, and it is debated whether clumps can survive energetic feedback from young stars, and afterwards migrate inwards to form galaxy bulges. Here we report the spatially resolved spectroscopy of a bright off-nuclear emission line region in a galaxy at z = 1.987. Although this region dominates star formation in the galaxy disk, its stellar continuum remains undetected in deep imaging, revealing an extremely young (less than ten million years old) massive clump, forming through the gravitational collapse of more than one billion solar masses of gas. Gas consumption in this young clump is more than tenfold faster than in the host galaxy, displaying high star-formation efficiency during this phase, in agreement with our hydrodynamic simulations. The frequency of older clumps with similar masses, coupled with our initial estimate of their formation rate (about 2.5 per billion years), supports long lifetimes (about 500 million years), favouring models in which clumps survive feedback and grow the bulges of present-day galaxies.

ICSH guidelines for the laboratory diagnosis of nonimmune hereditary red cell membrane disorders.

INTRODUCTION: Hereditary spherocytosis (HS), hereditary elliptocytosis (HE), and hereditary stomatocytosis (HSt) are inherited red cell disorders caused by defects in various membrane proteins. The heterogeneous clinical presentation, biochemical and genetic abnormalities in HS and HE have been well documented. The need to raise the awareness of HSt, albeit its much lower prevalence than HS, is due to the undesirable outcome of splenectomy in these patients. METHODS: The scope of this guideline is to identify the characteristic clinical features, the red cell parameters (including red cell morphology) for these red cell disorders associated, respectively, with defective cytoskeleton (HS and HE) and abnormal cation permeability in the lipid bilayer (HSt) of the red cell. The current screening tests for HS are described, and their limitations are highlighted. RESULTS: An appropriate diagnosis can often be made when the screening test result(s) is reviewed together with the patient's clinical/family history, blood count results, reticulocyte count, red cell morphology, and chemistry results. SDS-polyacrylamide gel electrophoresis of erythrocyte membrane proteins, monovalent cation flux measurement, and molecular analysis of membrane protein genes are specialist tests for further investigation. CONCLUSION: Specialist tests provide additional evidence in supporting the diagnosis and that will facilitate the management of the patient. In the case of a patient's clinical phenotype being more severe than the affected members within the immediate family, molecular testing of all family members is useful for confirming the diagnosis and allows an insight into the molecular basis of the abnormality such as a recessive mode of inheritance or a de novo mutation.

Hereditary red cell membrane disorders and laboratory diagnostic testing.

This overview describes two groups of nonimmune hereditary hemolytic anemias caused by defects in membrane proteins located in distinct layers of the red cell membrane. Hereditary spherocytosis (HS), hereditary elliptocytosis (HE), and hereditary pyropoikilocytosis (HPP) represent disorders of the red cell cytoskeleton. Hereditary stomatocytoses represents disorders of cation permeability in the red cell membrane. The current laboratory screening tests for HS are the osmotic fragility test, acid glycerol lysis time test (AGLT), cryohemolysis test, and eosin-5'-maleimide (EMA)-binding test. For atypical HS, SDS-polyacrylamide gel electrophoresis of erythrocyte membrane proteins is carried out to confirm the diagnosis. The diagnosis of HE/HPP is based on abnormal red cell morphology and the detection of protein 4.1R deficiency or spectrin variants using gel electrophoresis. None of screening tests can detect all HS cases. Some testing centers (a survey of 25 laboratories) use a combination of tests (e.g., AGLT and EMA). No specific screening test for hereditary stomatocytoses is available. The preliminary diagnosis is based on presenting a compensated hemolytic anemia, macrocytosis, and a temperature or time dependent pseudohyperkalemia in some patients. Both the EMA-binding test and the osmotic fragility test may help in differential diagnosis of HS and hereditary stomatocytosis.

Performance of different PEEP valves and helmet outlets at increasing gas flow rates: a bench top study.

BACKGROUND: Aim of the paper was to assess the performance of different expiratory valves and the resistance of helmet outlet ports at increasing gas flow rates. METHODS: A gas flow-meter was connected to 10 different expiratory peep valves: 1 water-seal valve, 4 precalibrated fixed PEEP valves and 5 adjustable PEEP valves. Three new valves of each brand, set at different pressure levels (5-7.5-10-12.5-15 cmH(2)O, if available), were tested at increasing gas flow rates (from 30 to 150 L/min). We measured the pressure generated just before the valves. Three different helmets sealed on a mock head were connected at the inlet port with a gas flow-meter while the outlet was left clear. We measured the pressure generated inside the helmet (due to the flow-resistance of the outlet port) at increasing gas flow rates. RESULTS: Adjustable valves showed a variable degree flow-dependency (increasing difference between the measured and the expected pressure at increasing flow rates), while pre-calibrated valves revealed a flow-independent behavior. Water seal valve showed low degree flow-dependency. The pressures generated by the outlet port of the tested helmets ranged from 0.02 to 2.29 cmH(2)O at the highest gas flow rate. CONCLUSION: Adjustable PEEP valves are not suggested for continuous-flow CPAP systems as their flow-dependency can lead to pressures higher than expected. Precalibrated and water seal valves exhibit the best performance. Different helmet outlet ports do not significantly affect the pressure generated during helmet CPAP. In order to avoid iatrogenic complications gas flow and pressure delivered during helmet CPAP must always be monitored.

Long-term extracorporeal membrane oxygenation with minimal ventilatory support: a new paradigm for severe ARDS?

Pulmonary tuberculosis can lead to acute respiratory distress syndrome (ARDS) which is associated with high mortality. We report the case of a patient with pulmonary tuberculosis and severe ARDS (PaO2/FiO2<100 mmHg) who was initially managed with advanced up-to-date treatments (protective ventilation and extracorporeal membrane oxygenation, ECMO) but failed to improve. After a month of failure and the development of bilateral pneumothoraces, we drastically changed our therapeutic strategy: we maximized ECMO support to maintain oxygenation, we greatly reduced ventilation pressures and we left the pneumothoraces undrained. From then on, the patient improved and he eventually survived. This case suggests that ECMO permits large reductions in lung inflation and ventilation to rest the lungs, while maintaining acceptable oxygenation. The combination of ECMO and markedly attenuated ventilation strategy may be effective in cases of severe ARDS.

Two-dimensional and Doppler echocardiographic findings in healthy non-sedated red-eared slider terrapins (Trachemys scripta elegans).

Echocardiographic evaluation was performed in six healthy young adult non-sedated terrapins (Trachemys scripta elegans). The best imaging quality was obtained through the right cervical window. Base-apex inflow and outflow views were recorded, ventricular size, ventricular wall thickness and ventricular outflow tract were measured, and fractional shortening was calculated. Pulsed-wave Doppler interrogation enabled the diastolic biphasic atrio-ventricular flow and the systolic ventricular outflow patterns to be recorded. The following Doppler-derived functional parameters were calculated: early diastolic (E) and late diastolic (A) wave peak velocities, E/A ratio, ventricular outflow systolic peak and mean velocities and gradients, Velocity-Time Integral, acceleration and deceleration times, and Ejection Time. For each parameter the mean, standard deviation and 95% confidence interval were calculated. Echocardiography resulted as a useful and easy-to-perform diagnostic tool in this poorly known species that presents difficulties during evaluation.

Gentle vs. aversive handling of pregnant ewes: I. Maternal cortisol and behavior.

The aim of the experiment was to study the effects of aversive vs. gentle handling in late pregnancy on maternal behavior of ewes. Sixteen Norwegian Dala ewes bearing twins were subjected to 10 min of either gentle (GEN--soft talking and calm behavior) or aversive (AVS--swift movements and shouting) handling twice a day during the last 5 weeks of pregnancy. Salivary cortisol was recorded before and after treatments. The following behaviors were recorded post-partum in the ewes: grooming duration, number of vocalizations and in the lambs: number of vocalizations, latency and duration of standing, latency and duration in udder-directed position. The ability of the ewe to follow her lamb carried away by a human was scored on day 1 and 7. After the treatment sessions, cortisol levels tended to increase in AVS ewes but not GEN ewes. At parturition, AVS ewes groomed their offspring for a longer duration than GEN ewes. AVS lambs tended to be heavier than GEN lambs at 24 h of age. Follow Scores from GEN ewes were higher than for the AVS ewes at day one, but no difference between treatment groups was detected after one week. These results show that aversively treated ewes increased their grooming behavior towards their offspring, but that fear of humans disrupted their ability to follow their lambs closely when carried away by a human. We conclude that the type of handling of ewes during pregnancy may have some impact on important maternal behaviors.

Gentle vs. aversive handling of pregnant ewes: II. Physiology and behavior of the lambs.

We compared the effects of aversive and gentle handling in late pregnant ewes on fearfulness, heart rate variability and spatial learning in lambs. Twenty-four Norwegian-Dala ewes were studied. Ewes were subjected to gentle (i.e. soft talking and calm behavior) or aversive handling (i.e. swift movements and shouting) for 10 min twice a day during the last five weeks of pregnancy. Lambs from aversively (AVS) or gently (GEN) treated ewes were tested at 4 weeks of age. Lamb behavior was recorded during a) a human approach test, composed of 4 min of isolation and 4 min of exposure to an unfamiliar human, b) an umbrella startle test followed by 5-min recording, and c) two repetitions of a maze test. In addition, heart rate variability was recorded telemetrically before and after the human and startle tests. The baseline heart rate variability measures suggested a lower influence of vagal stimulation in AVS lambs. In the human approach test, AVS lambs vocalized and explored the environment less, and were slower to approach the human. They also tended to have higher flight distances during the startle test than the GEN lambs. The prenatal treatment had no significant effect in the maze test. In conclusion, we showed that aversive handling of pregnant ewes increased fearfulness and reduced vagal tone in their progeny compared to GEN lambs. These effects can have consequences for how lambs cope with rearing conditions.

Risk factors for skin lesions on the necks of Norwegian dairy cows.

Physical restrictions at the feed barrier may cause skin lesions, especially on the necks of cows. Thus, some aspects of the functionality of feed barrier design can be assessed by observing skin lesions on the necks of dairy cattle. In this study, 232 herds, mainly of the breed Norwegian Red (94%), were investigated between September 2006 and June 2007. The herds were categorized into 2 groups. One data set, based on 1,148 cows (115 herds), was from farms using feed barriers with post-and-rail designs (86 herds) and combinations of barriers (29 herds). The second data set, which included 1,168 cows (117 herds), was from farms using only vertical feed barriers such as tombstone, diagonal, and headlock. Skin lesions were observed on the necks of 21% of the 2,335 cows observed. Forty-two percent of cows had neck lesions in herds with post-and-rail barriers, whereas 4% of cows had neck lesions in herds with vertical barriers and 30% of cows had neck lesions in herds with a combination of both horizontal and vertical barriers. Alternating logistic regression was used to identify factors associated with skin lesions on the necks of cows. The estimates were transformed into odds ratios (OR). In herds in which post-and-rail design barriers were used, the following risk factors were associated with skin lesions: manger front lower than 63 cm versus higher (OR=3.6), no feed stalls versus feed stalls (OR=10.9), mixed rations versus no mixed ration (OR=3.8), cows later than 30 DIM versus earlier (OR=1.9), cows with low milk production versus those producing 1,000 kg higher (OR=1.4), feeding frequency of less than 3 times daily versus 3 and more times daily (OR=1.9), using only post-and-rail feed barriers (OR=1.8) versus using combinations, and farmer disagreement with the statement "animals experience physical pain as humans do" (OR=1.9) versus agreement. Diagonal barriers, categorized as vertical barriers, increased the association of skin lesions with increasing manger front heights. Feed barrier design was associated with skin lesions on the necks of cows, particularly for barriers of a post-and-rail design. To minimize the risk of neck lesions, the top rail height of post-and-rail barriers should be above 109 cm, especially for cows taller than 126 cm, and the manger front should not be lower than 63 cm. Feed stalls, feeding frequency of 3 or more times daily, and the feeding of unmixed rations were also identified as preventive factors.

Dairy farmer attitudes and empathy toward animals are associated with animal welfare indicators.

Attitudes and empathy of farmers influence human-animal interaction, thereby affecting their behavior toward animals. The goal was to investigate how measures of attitude and empathy toward animals were associated with animal welfare indicators such as milk yield, mastitis incidence, fertility index, and the prevalence of skin lesions on cows. To assess empathy toward animals, a photo-based pain assessment instrument was developed depicting various conditions that could be associated with some degree of pain in cattle and included questions aimed at assessing attitudes toward animals. Photos of painful conditions are useful in eliciting measurable empathic responses to pain in humans. A total of 221 farmers were sampled via e-mail and 154 responses were obtained. In the first analysis, farmers were categorized into 2 groups according to their agreement or disagreement with the attitude statement "animals experience physical pain as humans do." In the second analysis, farmers were assigned a median pain assessment score obtained from their estimates on the visual analog scale of 21 conditions assumed painful for cattle. In the third analysis, farmers were clustered in 3 groups according to their visual analog scale responses. Three conditions were ranked as the most painful: fracture of tuber coxae, dystocia, and serious mastitis. Farmers with positive attitudes toward animals scored 2 points higher on their empathy score compared with farmers with negative attitudes. Personal experience with each additional condition resulted in a 0.09 higher score. Cluster analysis revealed 3 groups. Farmers in group 3 had the highest median pain assessment score (6.7+/-0.2), indicating a high level of empathy and a positive attitude toward animals. They had the lowest prevalence of skin lesions over the carpus (24+/-6%) and the lowest milk production (6,705+/-202 kg). The complex associations between indicators of empathy and attitudes with relevant welfare outcomes suggest that competence building to safeguard animal welfare could benefit from including both attitudes and empathy in human-animal interactions studies.

Copper mobilization affected by weather conditions in a stormwater detention system receiving runoff waters from vineyard soils (Champagne, France).

Copper, a priority substance on the EU-Water Framework Directive list, is widely used to protect grapevines against fungus diseases. Many vineyards being located on steep slopes, large amounts of Cu could be discharged in downstream systems by runoff water. The efficiency of stormwater detention basins to retain copper in a vineyard catchment was estimated. Suspended solids, dissolved (Cu(diss)) and total Cu (Cu(tot)) concentrations were monitored in runoff water, upstream, into and downstream from a detention pond. Mean Cu(tot) concentrations in entering water was 53.6 microg/L whereas it never exceeded 2.4 microg/L in seepage. Cu(tot) concentrations in basin water (>100 microg/L in 24% of the samples) exceeded LC(50) values for several aquatic animals. Copper was principally sequestered by reduced compounds in the basin sediments (2/3 of Cu(tot)). Metal sequestration was reversible since sediment resuspension resulted in Cu remobilization. Wind velocity controlled resuspension, explained 70% of Cu(diss) variability and could help predicting Cu mobilization.