Optimization of morphine extraction method for the assay of its urinary 3-glucuronideconjuguate by gas chromatography-mass spectrometry.

In the field of doping, a great interest is carried for the analysis of morphine, a powerful narcotic analgesic opiate which use is prohibited during competitions. In order to confirm the abnormal analytical result in our anti-doping laboratory, a sensitive and selective gas chromatography-mass spectrometry (GC-MS) method was performed for the quantification of urinary morphine. As sample preparation is a key step for the determination of drugs in biological samples, the aim of this work consists of the optimization of the urinary human sample pretreatment conditions before quantification by GC/MS. Enzymatic hydrolysis associated with liquid-liquid extraction constitute the major pre-treatment steps. Our study has first focused on the optimization of the extraction solvents then to enzymatic hydrolysis which morphine is released from its glucuronide conjugated form. Onboard premiums, a study involving the effect of "amount of enzyme", "incubation temperature" and "duration of hydrolysis" was conducted. This univariate study has enabled us to evaluate the influence of each of these operating variables on the area ratio of morphine to the internal standard (Amorphine/AIS) response and to set the experimental fields for each one of them. Based on these results, an experimental design was established using the Box-Behnken model to determine, by multivariate analysis, the optimal operating conditions maximizing the "Amophine/AIS" response. After validation, the analysis of response surface makes it possible to set the optimum operating conditions, which the ratio "Amorphine/AIS" is maximized. The retained conditions for enzymatic hydrolysis are 160µl of Escherichia coli glucuronidase enzyme during 6hours of incubation at a temperature of 36°C. The solvent mixture Methyl-t-Butyl Ether/isopropanol (4:1, v/v) was selected since it has improved morphine extraction from the urinary matrix allowing a gain of 50% when compared to that used in our routine laboratory. Our developed extraction method can be successfully applied for our forensic anti-doping analysis of morphin in human sample urine.

[Congenital laryngeal cyst: an uncommon cause of stridor in neonates]. / Kyste laryngé congénital: une cause rare de stridor chez le nouveau-né

Laryngeal cyst is a rare but generally benign lesion in the larynx. It may cause stridor and life-threatening airway obstruction in early infancy. We report the case of a neonate who developed respiratory distress 24h after birth in relation with materno-fetal infection. Airway obstruction was then suspected because of stridor. A laryngeal mass was discovered at the time of intubation. The infant experienced a nosocomial infection and died on day 14 of life before any surgical treatment. Autopsy concluded in an epiglottic congenital laryngeal cyst.

[Congenital pulmonary lymphangiectasis: an unusual cause of respiratory distress in neonates]. / Lymphangiectasies pulmonaires congénitales: une cause rare de détresse respiratoire néonatale.

Congenital pulmonary lymphangiectasis (CPL) is a rare condition in neonates characterized by abnormal dilatation of the lymphatics draining the interstitial and subpleural spaces of the lungs. Diagnosis is difficult in the neonatal period because respiratory features and radiological findings are not specific of the disease. Definitive diagnosis of CPL can be made only by pathologic examinations. We report a case of a male near-term neonate presenting with severe respiratory distress at birth. The initial chest X-ray showed frosted glass-like infiltrates with air bronchogram suggesting a maternofetal infection or respiratory distress syndrome. The infant required mechanical ventilation and chest tube insertion for right then bilateral pneumothorax. The child died 15 days later in spite of optimal high-frequency ventilation, bilateral pneumothorax drainage, and hemodynamic support. Autopsy revealed features consistent with the diagnosis of CPL. Although CPL is very rare, we should be aware that it is a possible cause of severe unexplained respiratory distress in neonates.

[Spontaneous neonatal arterial thrombosis: a report of 4 neonates]. / Thromboses artérielles néonatales non causées par un cathétérisme artériel: à propos de 4 observations.

Arterial thromboses are rare in newborns, mostly caused by arterial catheters. Neonatal arterial thrombosis occurring in other contexts is much rare. We report arterial thrombosis, not caused by catheterisation in 4 neonates hospitalised in the neonatology department of the Farhat Hached de Sousse hospital in Tunisia. The diagnosis of arterial thrombosis was made based on signs of ischemia in 2 patients affected by thrombosis of the iliac arteries. The symptoms were less clear: anuria associated with arterial high blood pressure in 1 patient affected by thrombosis of the abdominal aorta and by anuria with melena in a newborn with aortic and mesenteric thrombosis. Diagnosis was confirmed by Doppler sonography in 3 patients and based on autopsy data in 1 patient. A delay to consultation was noted in 3 patients, whose outcome was fatal. The progression was favourable after thrombolysis and anticoagulation using heparin in 1 patient with major aortic thrombosis. A review of the literature on the epidemiological, clinical, therapeutic and outcome data of the arterial thrombosis in the newborn child is provided.

[Thoracic congenital ectopic right kidney with diaphragmatic hernia: a rare developmental anomaly]. / Rein droit intrathoracique et hernie diaphragmatique congénitale : une association rare chez le nouveau-né

Ectopic intrathoracic kidney is a very rare congenital anomaly, which is often asymptomatic and discovered incidentally on chest radiography. Diagnosis of this ectopia in the neonatal period is extremely rare. We report the case of a female infant admitted for mild respiratory distress at birth in relation with maternofetal infection in whom a right posterior opacity on chest x-ray was diagnosed as intrathoracic kidney by sonography and chest CT scan. Diaphragmatic hernia could not be confirmed at this stage. At 2 months, the presence of air blebs on chest x-ray performed for viral bronchiolitis confirmed an associated right Bochdalek hernia. The infant, although asymptomatic, underwent surgery. The intrathoracic kidney was reduced into the abdominal cavity at the time of surgery. The postoperative course was uneventful and renal function was normal.

[Neonatal seizures revealing incontinentia pigmenti]. / Convulsions néonatales révélant une incontinentia pigmenti.

Incontinentia pigmenti (IP) is a rare affection inherited as X-linked dominant disease. It is usually lethal in male infants. IP can affect ectodermal tissues such as the skin, teeth, eyes, bones, and the central nervous system. Skin lesions occur mostly during the neonatal period and are characterized by a classic progression in 4 stages leading to hyperpigmentation. We report on the case of a female neonate presenting on the 3rd day of life with seizures without obvious cause, in which the diagnosis of IP was made 1 week later when skin lesions appeared.

[Systemic congenital candidiasis, a rare condition in neonates: case report in a premature infant]. / Candidose congénitale systémique, forme rare de candidose néonatale : à propos d'une observation chez un nouveau-né prématuré

UNLABELLED: Congenital cutaneous candidiasis presenting at birth is very uncommon and is due to intra-uterine infection. The systemic form has to be considered when cutaneous signs are associated with sepsis symptoms. CASE REPORT: A female infant was born by vaginal delivery at 35 weeks' gestation to a mother whose pregnancy had been complicated by urinary tract infection treated 3 days before delivery. The infant was admitted because of respiratory distress. Clinical features consisted of respiratory retraction signs associated with hepatomegaly and rash on the trunk. The white blood cell (WBC) count was 50 × 10(9)/L and C-reactive protein was negative. Maternofetal bacterial infection was suspected and intravenous antibiotics were prescribed. Over the next 6h, macules appeared on the trunk, back, and limbs, which changed after 24h into papulovesicular lesions over the trunk, back, limbs, palms, and scalp. Congenital candidiasis was suspected, confirmed by cultures from vesicle swabs and maternal vaginal discharge. The systemic form was considered because of respiratory distress requiring oxygen therapy for 4 days, hepatomegaly, elevated WBC count, and chest X-ray infiltrates. The infant was started on intravenous systemic antifungal therapy (fluconazole, 6 mg/day). Treatment was continued for 3 weeks. The rash resolved by desquamation after about 1 week and hepatomegaly disappeared. The infant remained well at follow-up.