Optimization of embryonic thermal programming confirms increased liver fattening in mule ducks and changes in lipid metabolism.

Introduction: The embryonic thermal programming (TM) in birds has been shown to impact several physiological parameters such as resistance to thermal stress, muscle growth or immunity. In mule ducks, it has recently been shown that TM can induce metabolic programming resulting in increased liver weight and fat storage after overfeeding. However, a decrease in hatchability and foie gras quality was also observed, suggesting that this technique needs to be optimized. Here, we tested a new thermal manipulation condition determined with the objective of avoiding negative impacts while maintaining or improving liver properties. Methods: The eggs of the control group were incubated at 37.6°C during the whole incubation period while those of the experimental group (TM group) were incubated at 39.3°C 16 h/24 h from the 11th day of incubation to the 21st. After hatching, all the animals were fed and raised under the same conditions until the age of 12 weeks. At this stage, one part of the animals was overfed and then slaughtered 2 h (to measure rapid changes in metabolism) or 10 h after the last meal (to obtain the best technological yields), while the other part was ration-fed and slaughtered 2 h after the last meal, at the same age. Results: An 8% increase in foie gras production was measured in the TM group compared to the control group without altering the quality of the final product (nor hatchability), confirming the successful optimization of the metabolic programming. Interestingly, these results allowed us not to reject the previously suggested hypothesis of a potential delay in metabolic processes involved in liver fattening in programmed animals, in particular by measuring a trend reversal regarding the amount of total hepatic lipids in both groups at 2 h and then 10 h after the last meal. Discussion: This study therefore validates the optimization of metabolic programming by embryonic thermal manipulation for duck liver fattening. The understanding of the mechanisms of embryonic thermal programming in birds remains today very incomplete and the search for epigenetic marks (main hypothesis of the concept of programming) at the origin of the observed phenotypes could be the next step of this work.

[Crossed eyes on arterial blood pressure in sitting position]. / L'image du mois. Regards croisés sur la pression artérielle en position assise.

Prolonged standing or even sitting position can induce a fall in arterial blood pressure because of gravity that provokes a blood volume sequestration in lower limbs. Such a phenomenon may be associated with orthostatic dizziness or sometimes syncope. We present the image of a continuous monitoring using a Finometer® of systolic blood pressure and heart rate. Crossing legs while in sitting position allows immediate increase of arterial blood pressure. This pathophysiological mechanism explains why numerous people cross legs in sitting position, for instance during a prolonged meeting, in an auditorium or in a movie theater. This common phenomenon has potential diagnostic, therapeutic and prognostic implications.

La position debout, ou même assise, prolongée peut entraîner une baisse de la pression artérielle compte tenu d'une certaine séquestration du volume sanguin dans les membres inférieurs due à la gravité, ce qui peut provoquer un malaise lipothymique, voire une syncope. Nous présentons l'image d'un enregistrement continu, avec un Finometer®, de la pression artérielle systolique et de la fréquence cardiaque, démontrant que le simple fait de croiser les jambes en position assise permet de faire remonter, quasi instantanément, la pression artérielle. Ce mécanisme physiopathologique explique pourquoi de nombreuses personnes croisent les jambes en position assise, que ce soit, par exemple, lors d'une réunion prolongée, dans un auditoire ou dans une salle de cinéma. Ce phénomène, a priori banal, a potentiellement des implications diagnostiques, thérapeutiques et pronostiques. Mots-clés : Pression artérielle - Hypotension - Position corporelle - Mesure continue - Finometer®.

Clinical Pharmacokinetics and Pharmacodynamics of Atezolizumab in Metastatic Urothelial Carcinoma.

Atezolizumab, a humanized immunoglobulin G1 (IgG1) monoclonal antibody targeting human programmed death-ligand 1 (PD-L1), is US Food and Drug Administration (FDA) approved in metastatic urothelial carcinoma (MUC) and is being investigated in various malignancies. This analysis based upon 906 patients from two phase I and one phase II MUC studies, is the first report of the clinical pharmacokinetics (PK) and pharmacodynamics (PD) of atezolizumab. Atezolizumab exhibited linear PK over a dose range of 1-20 mg/kg, including the labeled 1,200 mg dose. The clearance, volume of distribution, and terminal half-life estimates from population pharmacokinetic (PopPK) analysis of 0.200 L/day, 6.91 L, and 27 days, respectively, were as expected for an IgG1. Exposure-response analyses did not identify statistically significant relationships with either objective response rate or adverse events of grades 3-5 or of special interest. None of the statistically significant covariates from PopPK (body weight, gender, antitherapeutic antibody, albumin, and tumor burden) would require dose adjustment.

Susceptibility to acidification of groundwater-dependent wetlands affected by water level declines, and potential risk to an early-breeding amphibian species.

Eggs of the Western spadefoot toad (Pelobates cultripes) reached a 100% mortality in all 29 clutches deposited at a pH below 5.0 in a temporary pond of the Doñana National Park (SW Spain) throughout the wet season of 2006-2007. A similar trend was detected in a neighbouring pond. The proximity of these two ponds to a groundwater pumping area (<1.5km), prompted us to elucidate the possible links between the reduction in pond hydroperiod over past decades (1989-2008) and the decrease of groundwater pH-buffering capacity. The average hydroperiod had decreased by 4months since 1998-99 in the pond where the extensive egg mortality had occurred. The total alkalinity, and the Mg(2+)concentration had also significantly declined in the shallow water-table since 1998-99, from an average of 8.56 to 0.32meql(-1), and of 3.57 to 1.15meql(-1), respectively. This decline of the shallow groundwater buffering capacity could turn this pond more susceptible to the inorganic acidity associated with pyrite oxidation as the sediment S content was often above 0.03%. The initial ratio of S/Ca+Mg in the summer dry sediment was a good predictor of pore-water pH on re-wetting after desiccation (r(2)=0.802, p<0.01). Therefore, this ratio can give some anticipation to mitigate the impact of acidity on toad hatching before these temporary ponds are reflooded on the next wet season. Our results suggest that the long-term damage to pond water levels can trigger a potential risk of soil acidification in the presence of iron-sulphide minerals.

Early psychological care of the French victims of the Costa Concordia shipwreck.

Most of the French passengers who survived the shipwreck of the cruise ship Costa Concordia were repatriatedfrom Italy to Marseille, one of the stopovers of the cruise. The shipwreck happened during the nightof 13th-14th January 2012 and entailed the forced evacuation of 4195 passengers and crewmembers.Thirty-two persons died and 2 others are still reported missing. The massive and unexpected inflow of402 French citizens in the port of Marseille required the quick setting up of welcome facilities, not only tosolve logistical problems, but also to address psychological and sometimes even medical problems. ThePrehospital Psychological Emergency Service (CUMP) and the Prehospital Emergency Medical Service(SAMU) of Marseille examined 196 persons in total, and were able to avoid a great number of emergencyadmissions deemed necessary because of difficult psychological situations (death, missing or lost persons,acute stress). The objective of this report is to rapidly present the emergency committee as a whole andto describe in more detail the work that the CUMP accomplished during the 36 hours necessary to takecharge of the majority of the French passengers of the Costa Concordia.

[Psychiatric disorders associated with montelukast: data from the National Pharmacovigilance Database]. / Troubles psychiatriques associés au montélukast : données de la base nationale de pharmacovigilance.

Montelukast (Singulair(®)) has been the subject of post-marketing warnings about psychiatric events occurring that had not been identified during clinical trials. The objective of this study was to take stock of the adverse events (AEs) related to montelukast reported in France. Cases of psychiatric disorders reported to regional pharmacovigilance centers (CRPV) and the literature data were analyzed. The 56psychiatric AEs account for 20% of all AEs reported in the montelukast CRPV: essentially sleep disorders, behavioral disorders and depression. This risk is also found in pharmacovigilance databases in other countries, especially in the North American database, which recorded a significant number of cases of "suicidality", including suicidal ideation, suicide attempts, and suicides. Analysis of clinical efficacy studies have failed to confirm these AEs. The potential severity of these events prompts physicians to seek the existence of psychiatric disorders before prescribing the drug and to carefully monitor the occurrence of AEs during treatment.

Substantiation of an artificial saliva formulated for use in a masticatory apparatus.

The aim of this work was to substantiate artificial saliva prepared for use in a masticator apparatus. Mastication's goal is to produce a viscous and plastic food bolus where these properties authorize a safe swallow. Apart from its biochemical contribution, saliva is mainly used in this kind of apparatus to provide a viscous component to the bolus. Artificial saliva was prepared with water and minerals, and completed with mucin and amylase. Different physico-chemical conditions were applied and the resultant viscosity was compared to that of human saliva. Mechanically- or chemically-stimulated salivas of ten healthy subjects were collected. Viscosity was measured with a capillary viscometer in response to changes in measurement's temperature, air exposure or pH. The effects of circadian saliva collection and the stimulation type on viscosity of human saliva were also studied. Viscosity of artificial and human salivas was comparable. An increase in the measurement's temperature or a 30 min-exposure of saliva to air led to a significant decrease in viscosity of both types of saliva. Amylase in artificial saliva did not change viscosity. The viscosity of human saliva displayed important subject variability as well as a dependence on the stimulation type of saliva production. This work allowed a useful evaluation of the formulated artificial saliva. It exhibited similar viscosity as the natural saliva in response to different methodological conditions. Therefore the proposed artificial saliva satisfies the major requirement of viscosity for a use in the masticator apparatus designed to prepare a food bolus.

[Pharmacovigilance in children]. / Pharmacovigilance en pédiatrie.

Drug safety in children must take into account the frequency of « off label ¼ prescriptions, children's growth dynamics, and possible long-term consequences (growth, neurodevelopment). The pharmacovigilance methodology is based on spontaneous notification and pharmacoepidemiology studies usually included the in risk management plan. Despite an increased drug risk (pharmacokinetic and pharmacodynamic specificities), drug safety is better in children than in adults. The incidence of drug side effects depends on the country, the type of study (in or out of the hospital), and age. Antibiotics, central nervous, respiratory and dermatologic drug systems are most often involved. The target organs are gastrointestinal and neurologic. In neonates, the most frequent side effects are due to pregnancy exposure to psychotropic drugs, beta-blockers, and antiepileptics. Some studies have shown an increased risk of off-label prescriptions in children. During the last 6 years in France, pediatric alerts (desmopressin, metoclopramide, bronchial mucolytic drugs, first-generation anti-H1, Uvesterol D(®), and Uvesterol A.D.E.C(®), rotavirus vaccines, growth hormone, cisapride) have been less frequent than in adults.

[Relationships between baroreflex gain and pulsatile stress in type 1 diabetic patients]. / Relations entre gain baroréflexe et stress pulsatile chez le patient diabétique de type 1.

AIM OF THE STUDY: Cardiovascular autonomic neuropathy (CAN) and early arterial stiffness are frequent complications in type 1 diabetes. The aim of our work is to study the relationships between CAN (estimated by baroreflex gain calculation) and arterial stiffness (estimated by pulsatile stress) in type 1 diabetic patients. PATIENTS AND METHODS: In a cross-sectional study, we calculated baroreflex gain and pulsatile stress in 167 type 1 diabetic patients and 160 matched non-diabetic subjects whose blood pressure was continuously monitored with a Finapres(®) device in a postural test (squatting test). The baroreflex gain was calculated by plotting the pulse intervals (R-R) against systolic blood pressure values during the transition phase from squatting to standing. Pulsatile stress was estimated by the pulse pressure×heart rate product. In a longitudinal study, the baroreflex gain and pulsatile stress were calculated before and after a mean follow-up of 79±33 months in type 1 diabetic patients. RESULTS: Cross-sectional data showed a decrease in baroreflex gain and an increase in pulsatile stress in type 1 diabetic patients versus the matched non-diabetic subjects. A significant correlation between the baroreflex gain and pulsatile stress was present. Type 1 diabetic patients with lower baroreflex gain had a higher value of pulsatile stress when compared to those with higher baroreflex gain. During follow-up, a significant reduction in baroreflex gain (but without significantly increased pulsatile stress) was observed. A univariate analysis showed that the decrease of the baroreflex gain is not correlated with the time interval between the two tests, neither type 1 diabetes duration nor mean glycated hemoglobin values, but significantly with the pulsatile stress increase. CONCLUSION: In type 1 diabetic patients, the baroreflex gain is decreased and the pulsatile stress is increased when these markers are compared to age-matched non-diabetic subjects. There is a relationship between indices of CAN and arterial stiffness. Nevertheless, the baroreflex gain (marker of CAN) is impaired earlier than the pulsatile stress in this type 1 diabetic population with inadequate glycaemic control.

Haemodynamic changes during a squat test, pulsatile stress and indices of cardiovascular autonomic neuropathy in patients with long-duration type 1 diabetes.

AIM: Cardiovascular autonomic neuropathy (CAN) and pulsatile stress are considered to be independent cardiovascular risk factors. This study compared haemodynamic changes during an active orthostatic test in adult patients with type 1 diabetes (T1DM), using low versus high RR E/I ratios as a marker of CAN. METHODS: A total of 20 T1DM patients with low RR E/I ratios were compared with 20 T1DM patients with normal RR E/I ratios, matched for gender (1/1 ratio), age (mean: 46 years) and diabetes duration (22-26 years); 40 matched healthy subjects served as controls. All subjects were evaluated by continuous monitoring of arterial blood pressure (Finapres(®)) and heart rate using a standardized posture test (1-min standing, 1-min squatting, 1-min standing), thus allowing calculation of baroreflex gain. RESULTS: Compared with controls, T1DM patients showed lower RR E/I ratios, reduced baroreflex gains, higher pulsatile stress (pulse pressure×heart rate), greater squatting-induced pulse pressure rises, orthostatic hypotension and reduced reflex tachycardia. Compared with T1DM patients with preserved RR E/I ratios, T1DM patients with low RR E/I ratios showed reduced post-standing reflex tachycardia and baroreflex gain, and delayed blood pressure recovery, but no markers of increased pulsatile stress. Interestingly, decreased baroreflex gain was significantly associated with both pulsatile stress and microalbuminuria. CONCLUSION: The use of RR E/I ratios to separate T1DM patients allows the detection of other CAN markers during an orthostatic posture test, but with no significant differences in pulsatile stress or microalbuminuria. In this context, squatting-derived baroreflex gain appears to be more informative.

Feature Selection with Conjunctions of Decision Stumps and Learning from Microarray Data.

One of the objectives of designing feature selection learning algorithms is to obtain classifiers that depend on a small number of attributes and have verifiable future performance guarantees. There are few, if any, approaches that successfully address the two goals simultaneously. To the best of our knowledge, such algorithms that give theoretical bounds on the future performance have not been proposed so far in the context of the classification of gene expression data. In this work, we investigate the premise of learning a conjunction (or disjunction) of decision stumps in Occam's Razor, Sample Compression, and PAC-Bayes learning settings for identifying a small subset of attributes that can be used to perform reliable classification tasks. We apply the proposed approaches for gene identification from DNA microarray data and compare our results to those of the well-known successful approaches proposed for the task. We show that our algorithm not only finds hypotheses with a much smaller number of genes while giving competitive classification accuracy but also having tight risk guarantees on future performance, unlike other approaches. The proposed approaches are general and extensible in terms of both designing novel algorithms and application to other domains.

Squatting, a posture test for studying cardiovascular autonomic neuropathy in diabetes.

Cardiovascular autonomic neuropathy (CAN) is a frequent complication of diabetes mellitus, which is associated with increased morbidity and mortality. It involves both the parasympathetic and sympathetic nervous systems, and may be diagnosed by classical dynamic tests with measurements of heart rate (HR) and/or arterial blood pressure (BP). An original squat test (1-min standing, 1-min squatting, 1-min standing) was used with continuous monitoring of HR and BP, using a Finapres(®) device. This active test imposes greater postural stress than the passive head-up tilt test, and provokes large changes in BP and HR that can be analyzed to derive indices of CAN. In healthy subjects, squatting is associated with BP increases and HR decreases (abolished by atropine: SqTv index), whereas the squat-stand transition is accompanied by a deep but transient drop in BP associated with sympathetic-driven tachycardia (abolished by propranolol: SqTs index). In diabetic patients with CAN, BP increases are accentuated during squatting whereas reflex bradycardia is reduced. When standing from squatting position, the fall in BP tends to be more pronounced and orthostatic hypotension more prolonged, while reflex tachycardia is markedly dampened. The baroreflex gain, similar to that calculated during pharmacological testing with vasodilator/vasopressor agents, can be derived by plotting pulse intervals (R-R) against systolic BP levels during the biphasic response following the squat-stand transition. The slope, which represents baroreflex sensitivity, is significantly reduced in patients with CAN. This discriminatory index allows study of the natural history of CAN in a large cohort of diabetic patients.

Changes in pulse pressure, heart rate and the pulse pressure x heart rate product during squatting in Type 1 diabetes according to age.

AIMS: We assessed changes in pulse pressure and heart rate during a squatting test, as indirect markers of arterial stiffness and cardiovascular autonomic neuropathy, respectively, according to age and sex in patients with Type 1 diabetes mellitus. METHODS: We evaluated 160 diabetic patients, divided into four groups of 20 men and 20 women according to age (G1, 20-30 years old; G2, 31-40 years old; G3, 41-50 years old; and G4, 51-60 years old), and 160 non-diabetic matched control subjects. Each subject underwent a 3 min posture test (standing-squatting-standing) with continuous measurement of arterial blood pressure and heart rate by a Finapres device. Overall values throughout the test, baseline levels in initial standing position and squatting-induced changes in pulse pressure, heart rate and the pulse pressure x heart rate product were compared between diabetic patients and healthy control subjects. RESULTS: In the standing position, a greater increase in pulse pressure and lower reduction in heart rate with age led to a significantly higher pulse pressure x heart rate product in diabetic patients compared with control subjects. In the squatting position, a more marked pulse pressure increase in the absence of appropriate reduction in heart rate resulted in a greater rise in the pulse pressure x heart rate product in diabetic patients than in healthy subjects. No major differences were noted between the sexes, with the exception of a stronger relationship between pulse pressure and age in the female population with diabetes. Squatting-derived indices of cardiovascular autonomic neuropathy were also noted with increasing age in diabetic patients. CONCLUSIONS: The marked increase in the pulse pressure x heart rate product ('pulsatile stress') according to age, combined with cardiovascular autonomic neuropathy, may contribute to the higher cardiovascular risk of patients with Type 1 diabetes.

Pancreatic hypoplasia presenting with neonatal diabetes mellitus in association with congenital heart defect and developmental delay.

Congenital pancreatic hypoplasia is a rare cause of neonatal diabetes. We report on a series of three patients with pancreatic agenesis and congenital heart defects. All had abdominal scan evidence of pancreatic agenesis. In addition, Patient 1 had a ventricular septal defect, patent ductus arteriosus and pulmonary artery stenosis; Patient 2 had a truncus arteriosus and Patient 3 had tetralogy of Fallot. Two of the three patients have developmental delay. All three patients were isolated cases within the family. Investigations included sequencing of GCK, ABCC8, IPF1, NEUROD1, PTF1A, HNF1B, INS, ISL1, NGN3, HHEX, G6PC2, TCF7L2, SOX4, FOXP3 (Patients 1 and 2), GATA4 and KCNJ11 genes (all three patients), but no mutations were found. Genetic investigation to exclude paternal UPD 6, methylation aberrations and duplications of 6q24 was also negative in all three. 22q11 deletion was excluded in all three patients. Array CGH in Patient (1) showed a approximately 250 kb, paternally inherited duplication of chromosome 12q [arr cgh 12q24.33 (B35:CHR12:131808577-132057649++) pat], not found in the other two patients. Permanent neonatal diabetes mellitus due to pancreatic hypoplasia with congenital heart defects has been reported before and may represent a distinct condition. We discuss this rare association and review previously reported literature.

Supporting the recommended paediatric dosing regimen for rufinamide in Lennox-Gastaut syndrome using clinical trial simulation.

Rufinamide was approved for the treatment of seizures associated with Lennox-Gastaut syndrome (LGS) as adjunctive therapy in patients aged 4 years and older. Rufinamide pharmacokinetics (PK) has been established on pooled data from several clinical studies in epilepsy, including one in LGS patients. Demographic covariates and drug-drug interactions with several antiepileptic drugs have been explored using population PK modelling. Two types of drug-drug interactions models were developed and compared. The PK analysis demonstrated that the coadministration of valproate decreases rufinamide clearance, requiring potential dose adjustment. To explore rufinamide exposure under different dosing regimens in LGS patients, clinical trial simulations were performed. The objective of the simulations was to select the doses giving an exposure shown to be safe and efficacious in larger populations. The concentrations simulated in a subgroup of patients with body weight less than 30 kg presented a larger inter-individual variability than in other patients. Additional simulations demonstrated that this increased variability was due partly to greater valproate concentrations in some of the children treated with rufinamide. Simulations of the rufinamide exposure under different maximum daily dose in presence and in absence of valproate co-administration were used to establish the dosing recommendation. The simulations support the proposal of a lower maximum daily rufinamide dose for patients under 30 kg receiving both drugs: the dose of 600 mg/day was proposed as a maximum daily dose in children also receiving valproate concomitantly, whereas in absence of valproate, the maximum daily dose is 1000 mg/day.

Pulse pressure and cardiovascular autonomic neuropathy according to duration of type 1 diabetes.

BACKGROUND: To evaluate changes in pulse pressure (PP) and markers of cardiovascular autonomic neuropathy (CAN) according to duration of type 1 diabetes mellitus (T1DM). METHODS: This cross-sectional controlled study evaluated 159 diabetic patients during a 3-min posture test (standing-squatting-standing) with continuous measurement of systolic (SBP), diastolic (DBP) and mean (MBP) blood pressure by a Finapres device. Arterial stiffness was indirectly assessed by PP and the slope of PP as a function of MBP calculated during the whole 3-min test. CAN was assessed by the expiration/inspiration pulse interval ratio (E/I R-R ratio) during deep breathing and by three indices measured during the squatting test. Patients were divided into four groups according to diabetes duration (<10 years, 11-20 years, 21-30 years and > 30 years from group 1 to group 4, respectively) and compared with age-matched non-diabetic subjects. RESULTS: PP progressively increased (p < 0.0001) and PP/MBP decreased (p < 0.0005) according to T1DM duration, whereas these parameters remained almost unchanged in age-matched control subjects. E/I ratio (p < 0.0001) and baroreflex gain (p < 0.0005) progressively decreased with T1DM duration. The parasympathetic index (squatting test vagal ratio-SqTv) significantly increased (p < 0.0001), whereas the sympathetic index (squatting test sympathetic ratio-SqTs) only tended to decrease (p = 0.12) according to diabetes duration. No such changes in CAN indices were observed in the non-diabetic population. CONCLUSIONS: PP increased according to T1DM duration in an age range where PP remained almost stable in controls, in agreement with accelerated arterial stiffening due to chronic hyperglycaemia. The baroreflex gain decreased and other indices of CAN also deteriorated with diabetes duration, more so indices reflecting parasympathetic rather than sympathetic dysfunction.

Histopathological alterations in the liver of the sharptooth catfish Clarias gariepinus from polluted aquatic systems in South Africa.

There is a need for sensitive bio-monitoring tools in toxicant impact assessment to indicate the effect of toxicants on fish health in polluted aquatic ecosystems. Histopathological assessment of fish tissue allows for early warning signs of disease and detection of long-term injury in cells, tissues, or organs. The aim of this study was to assess the degree of histopathological alterations in the liver of C. gariepinus from two dams in an urban nature reserve, (Gauteng, South Africa). Two dams (Dam 1 and Dam 2) were chosen for their suspected levels of toxicants. Water and sediments were sampled for metal and potential endocrine disrupting chemical analysis. A quantitative and qualitative histology-based health assessment protocol was employed to determine the adverse health effects in fish. The analysis of blood constituents, fish necropsy, calculation of condition factors, and hepatosomatic indices were employed to support the findings of the qualitative and quantitative histological assessment of liver tissue. Assessment of the liver tissue revealed marked histopathological alterations including: structural alterations (hepatic cord disarray) affecting 27% of field specimens; plasma alterations (granular degeneration 98% and fatty degeneration 25%) of hepatocytes; an increase in melanomacrophage centers (32%); hepatocyte nuclear alterations (90%); and necrosis of liver tissue (14%). The quantitative histological assessment indicated that livers of fish collected from Dam 1 were more affected than the fish livers collected from Dam 2.

[Sweat testing: review of technical requirements]. / Recommandations pour l'exécution et l'interprétation du test de la sueur.

The sweat test, a quantitative measurement of chloride in sweat, remains a key laboratory test to support the diagnosis of cystic fibrosis. However, because of its delicate execution, sweat test result should be interpreted with biological, clinical and genetic arguments. The following guidelines which we propose, were established in order to harmonize the practices of the sweat test. They are elaborated in a consensual way by biologists from cystic fibrosis reference centers and/or from the working group "Sweat Testing" of the National College of Biochemistry Hospital praticiens, according to the current state of knowledge on the subject, the experiment of the biologists and the recommendations established in the United States and in the United Kingdom.