Association study of candidate DNA-repair gene variants and acute graft versus host disease in pediatric patients receiving allogeneic hematopoietic stem-cell transplantation.

Acute Graft versus Host Disease (aGvHD) grades 2-4 occurs in 15-60% of pediatric patients undergoing allogeneic haematopoietic stem-cell transplantation (allo-HSCT). The collateral damage to normal tissue by conditioning regimens administered prior to allo-HSCT serve as an initial trigger for aGvHD. DNA-repair mechanisms may play an important role in mitigating this initial damage, and so the variants in corresponding DNA-repair protein-coding genes via affecting their quantity and/or function. We explored 51 variants within 17 DNA-repair genes for their association with aGvHD grades 2-4 in 60 pediatric patients. The cumulative incidence of aGvHD 2-4 was 12% (n = 7) in the exploratory cohort. MGMT rs10764881 (G>A) and EXO rs9350 (c.2270C>T) variants were associated with aGvHD 2-4 [Odds ratios = 14.8 (0 events out of 40 in rs10764881 GG group) and 11.5 (95% CI: 2.3-191.8), respectively, multiple testing corrected p ≤ 0.001]. Upon evaluation in an extended cohort (n = 182) with an incidence of aGvHD 2-4 of 22% (n = 40), only MGMT rs10764881 (G>A) remained significant (adjusted HR = 2.05 [95% CI: 1.06-3.94]; p = 0.03) in the presence of other clinical risk factors. Higher MGMT expression was seen in GG carriers for rs10764881 and was associated with higher IC50 of Busulfan in lymphoblastoid cells. MGMT rs10764881 carrier status could predict aGvHD occurrence in pediatric patients undergoing allo-HSCT.

Antioxidant and antifungal activities of marrubiin, extracts and essential oil from Marrubium vulgare L. against pathogenic dermatophyte strains.

OBJECTIVE: Medicinal plants extracts and plant-derived compounds are one of the natural sources for discovering new antifungal agents, the objectives of this work were to investigate for the first time the antidermatophytic, antipathogenic activities of methanol, acetone extracts, and essential oil of Marrubium vulgare L. grown in Tunisia and its active compound marrubiin on pathogenic for animals and humans, such as some dermatophytes and pathogenic for plants, and to evaluate antioxidant activities of different extracts with consideration to their chemical compositions. MATERIAL AND METHODS: Acetone and methanol extracts were evaluated by HPLC, the essential oil was also analyzed by GC/MS. PCL assay was used to determine the antioxidant activity. RESULTS: Results showed that methanol and acetone extracts exhibited a significant antioxidant activity (261.41 and 272.90µmol TE/g respectively), while the lowest one was observed in the case of marrubiin and essential oil. The antifungal activity of different extracts, marrubiin and essential oil at two concentrations (20 and 100µg/mL) were screened against the dermatophytes fungi Microsporum gypseum, Microsporum canis, Arthroderma cajetani, Trichophyton mentagrophytes, Trichophyton tonsurans, Epidermophyton floccosum and against two fungi strains (Botrytis cinerea, Pythium ultimum). Among tested extracts, marrubiin at 100µg/mL showed about 50% inhibition for T. mentagrophytes and E. floccosum. The anti-phytopathogenic activity was also carried out, only marrubiin had in activity against B. cinerea at the highest dose (32.40%), while methanol extract of M.vulgare and marrubiin are able to increase the mycelial growth of P. ultimum at the highest concentration (45.15 and 40.30% respectively). CONCLUSION: In our study, we conclude that M.vulgare and marrubiin can be used as natural antioxidants and antifungal agent for treatment of skin dermatophyte infections.

Association of CTH variant with sinusoidal obstruction syndrome in children receiving intravenous busulfan and cyclophosphamide before hematopoietic stem cell transplantation.

Sinusoidal obstruction syndrome (SOS) is a severe complication of hematopoietic stem cell transplantation (HSCT) that can be fatal, often attributed to the conditioning regimen prior to HSCT. We evaluated the association of SOS risk with gene variants in cystathionase (CTH), an enzyme involved in glutathione synthesis, in 76 children receiving intravenous busulfan (Bu) before HSCT. Our results indicated an association with CTHc.1364 G>T (ORTT=10.6, 95% confidence interval (CI)=2.16, 51.54) and SOS risk, which was sex dependent (female patients, ORTT=21.82, 95% CI=3.590-132.649). The interaction between CTHc.1364 G>T and another risk variant (GSTA1*B) was explored. A recessive model with the use of GSTA1*B*B and CTH c.1364 TT genotypes proved to be useful at predicting SOS occurrence, indicating the possibility of using these gene variants as markers of SOS occurrence and to further individualize preemptive treatment aimed at reducing SOS incidence.

Influence of glutathione S-transferase gene polymorphisms on busulfan pharmacokinetics and outcome of hematopoietic stem-cell transplantation in thalassemia pediatric patients.

Hematopoietic stem-cell transplantation (HSCT) is currently the only curative therapeutic option for the treatment of thalassemia. In spite of the high cure rate, HSCT can lead to life-threatening adverse events in some patients. Busulfan (Bu) is a key component of the conditioning regimen prior to HSCT. Inter-individual differences in Bu pharmacokinetics (PK) are hypothesized to influence Bu efficacy and toxicity. Since Bu is mainly metabolized by glutathione S-transferase (GST), we investigated the relationship of GSTA1 and GSTM1 genotypes with first-dose PK and HSCT outcomes in 44 children with thalassemia intermedia and thalassemia major. All children received a myeloablative conditioning regimen with IV Bu. Association analysis revealed a relationship between GSTA169C>T (or haplotype *A/*B) and first Bu dose PK that was dependent on sex and Pesaro risk classification (PRC). Among female patients and patients with PRC I-II, homozygous individuals for the GSTA1T-69 allele defining haplotype *B, had higher Bu exposure and lower clearance (P⩽0.01). Association with HSCT outcomes showed that patients with the GSTM1 null genotypes had higher occurrence of regimen-related toxicity (P=0.01). These results suggest that GST genotypes could be useful to tailor the first Bu dose accordingly to improve HSCT outcome.

The association of cytochrome P450 genetic polymorphisms with sulfolane formation and the efficacy of a busulfan-based conditioning regimen in pediatric patients undergoing hematopoietic stem cell transplantation.

Cytochrome P450 enzymes (CYPs) and flavin-containing monooxygenases (FMOs) likely have a role in the oxidation of intermediate metabolites of busulfan (Bu). In vitro studies to investigate the involvement of these enzymes are cumbersome because of the volatile nature of the intermediate metabolite tetrahydrothiophene (THT) and the lack of sensitive quantitation methods. This study explored the association between the CYP2C9, CYP2C19, CYP2B6 and FMO3 genotypes and sulfolane (Su, a water soluble metabolite of Bu) plasma levels in children undergoing hematopoietic stem cell transplantation (HSCT). The relationship between these genotypes and the effectiveness of myeloablative conditioning was also analyzed. Sixty-six children receiving an intravenous Bu-based myeloablative conditioning regimen were genotyped for common functional variant alleles in CYP2C9 (*2 and *3), CYP2C19 (*2 and *17), FMO3 (rs2266780, rs2266782 and rs1736557) and CYP2B6 (*5 and *9). The plasma levels of Bu and its metabolite Su were measured after the ninth Bu dose in a subset of 44 patients for whom plasma samples were available. The ratio of Bu to Su was considered the metabolic ratio (MR) and was compared across the genotype groups. Higher MRs were observed in CYP2C9*2 and *3 allele carriers (mean±s.d.: 7.8±3.6 in carriers vs 4.4±2.2 in non-carriers; P=0.003). An increased incidence of graft failure was observed among patients with an MR>5 compared with those with MR values <5 (20% vs 0%; P=0.02). In contrast, a significantly higher incidence of relapse and graft failure (evaluated as event-free survival) was observed in patients with malignant disease who carried CYP2B6 alleles with reduced function on both chromosomes compared with carriers of at least one normal allele (100% vs 40%; P=0.0001). These results suggest that CYP2C9 has a role in the oxidation reactions of THT and indicate that it may be possible to predict the efficacy of Bu-based myeloablative conditioning before HSCT on the basis of CYP genotypes and Bu MRs.

Periodic variation of the water use efficiency in durum wheat.

Water Use Efficiency (WUF) of five durum wheat varieties (Khiar, Karim, Nasr, Razzak and Sobirano) cultivated in Tunisia was investigated. The experiment was conducted under rainfed field conditions, irrigated pots and test tubes at ESA Kef experiment station during the cropping season of 2010-2011. Under rainfed conditions, Khiar has the highest grain yield with 29.35 q ha(-1). Biological WUE ranged from 1.72-1.49 mg dry matter per g of water for Razzak and Nasr, respectively. The WUE of grain ranged.from 0.81-0.48 mg grain per g of water for Khiar and Sobirano, respectively. Biological WUE showed positive correlation to grain yield and stomatal density and negatively correlated with leaf area. Periodic water use efficiency in pot and test tube trials indicated variation of wheat genotypes according to stage periods and how the experiment was conducted. Test tubes trials showed high, accurate and maximum value of water use. Therefore, we can consider that Khiar has a high WUE in the first period; germination-early tillering (6.70 mg dry matter per g water). Nasr and Sobirano valorise well had got water use during the second period; tillering (9.16 and 10.20 mg dry matter per g of water). Razzak shows a high WUE for the third period; booting-early heading (9.06 mg dry matter per g of water). Finally, Karim cultivar has the highest WUE during for the fourth period; heading-physiological maturity (22.13 mg dry matter per g of water). Periods of high WUE can be targeted for appropriate time of wheat cultivar irrigation.

Glutathione S-transferase gene variations influence BU pharmacokinetics and outcome of hematopoietic SCT in pediatric patients.

BU is a key compound of conditioning regimens in children undergoing hematopoietic SCT (HSCT). Inter-individual differences in BU pharmacokinetics (PKs) might affect BU efficacy and toxicity. As BU is mainly metabolized by glutathione S-transferase (GST), we investigated the relationship between GSTA1, GSTM1 and GSTP1 genotypes with first-dose BU PKs, and the relationship with HSCT outcomes in 69 children receiving myeloablative conditioning regimen. GSTM1 null genotype correlated with higher BU exposure and lower clearance in patients older than 4 years (P ≤ 0.04). In accordance with the suggested functional role, GSTA1*A2 haplotype was associated with lower drug levels and higher drug clearance (P ≤ 0.03). Gene-dosage effect was also observed (P ≤ 0.007). GSTA1 haplotypes were associated with HSCT outcomes. Patients with two copies of haplotype *A2 had better event free survival (P=0.03). In contrast, homozygous individuals for haplotypes *B and *B1 had higher occurrence of veno-occlusive disease (P=0.009). GSTM1 null individuals older than 4 years had more frequently graft versus host disease (P=0.03). In conclusion, we showed that GST gene variants influence BU PK and outcomes of HSCT in children. A model for the dosage adjustment with the inclusion of genetic and non-genetic factors should be evaluated in a future prospective validation cohort.

[Calcification following intradiscal injection, a continuing problem?]. / Calcifications post-nucléorthèse, un problème toujours d'actualité?

INTRODUCTION: Intradiscal injection of triamcinolone hexacetonide, used to treat sciatica caused by herniated discs was discontinued after discovery that it was followed in some cases by epidural calcification. OBSERVATION: Ten years after an intradiscal injection at L4-L5, a 40 year-old man developed voluminous perivertebral, bilateral foraminal and intraductal calcifications that compressed the dural sheath. After preliminary excision, these calcifications recurred and required further excision, followed by posterior lateral grafting and osteosynthesis. DISCUSSION: Calcifications following intradiscal injections are symptomatic in 14 to 68% of cases. They require radical surgical treatment to avoid recurrence.

[Evaluation of outcome of patients hospitalized after pre-hospital cardiac arrest]. / Evaluation des patients hospitalisés au décours d'un arrêt cardiaque extrahospitalier.

The aim of this study was to assess management of patients resuscitated after pre-hospital cardiac arrest, initially indicated to preserve neurological status, the aetiological investigation only being undertaken when the outcome is favourable. Eighty-nine pre-hospital cardiac arrests were analysed retrospectively. The hospital survival was 16%, death being due to neurological lesions (55%), uncontrollable haemodynamic instability -39%) or other causes (7%). One year after the initial episode, none of the survivors had died, all living autonomously without (8 patients) or with minimal neurological sequellae (5 patients). These results are concordant with reports in the literature. The 11 cases of cardiac arrest with a favourable outcome of presumed cardiac origin underwent coronary angiography (6 cases) or endocavitary electrophysiological investigation (8 cases). These investigations showed or suggested an ischaemic process in 4 cases, an arrhythmia in 6 cases and severe valvular heart disease in 1 case. The independent predictive factors of survival were a Glasgow score of 6 or more on admission, the persistence of a light reflex and benign EEG appearances according to Synek's classification. The authors conclude that these results are comparable to those reported in the literature with aetiological investigations reserved for cases of favourable neurological outcome. The investigations including coronary angiography and electrophysiological investigation are essential as shown by the diversity of the cardiac pathologies identified.

[Long-term survival after extra-hospital cardiac arrest]. / Survie à long terme après arrêt cardiaque extra-hospitalier.

Survival rate after out-of-hospital cardiac arrest varies according to evaluation criteria. It can be estimated that in 22 to 63% of the cases, effective hemodynamic performance is restored although hospital mortality is much higher, reaching 63%. Death, frequent after prolonged cardiac arrest, is usually due to recurrent cardiac arrest or the effects of prolonged anoxia. Mortality in patients who survive the hospitalization period is approximately 20% during the year following discharge. Consequently one year after out-of-hospital cardiac arrest, only 5% of the patients are still alive. The quality of life varies greatly in these survivors; the course of neurological sequellae may be favorable in approximately half but leads to death in others. The primary factor predicting survival is the underlying pathology, highly influenced by age. Inversely, factors predicting a more favorable outcome include ventricular tachycardia as the origin of cardiac arrest, presence of other people at onset and rapid recovery of spontaneous hemodynamic activity. Loss of consciousness for more than 24 hours, defective bulbar reflexes and anomalies on the electroencephalogram are signs of gravity as are high blood glucose, major brain edema and abolition of somesthesic and auditive evoked potentials.

[Maternal mortality at the Maternity and Neonatology Center in Rabta of Tunis from 1986 to 1989]. / La mortalité maternelle au Centre de Maternité et de Néonatologie de la Rabta de Tunis de 1986 à 1989.

We studied 29 cases of maternal death occurring over a period of 3 years in the "Centre de Maternité et de Néonatologie de La Rabta-Tunis"; managed by the same staff. 42,028 live births occurred during the study years with 43,220 total births from April 24th 1986 to April 23rd 1989. The maternal mortality rate was 69 per 100,000 live births. A maternal age of under 35, nulliparity and grand multiparity were found, as is well known, to be risk factors. Maternal transfer in obstructed labour from rural maternity units raises the maternal death risk 12 times: 14 deaths out of 29 occurred in transferred patients. Haemorrhage represents a quarter of the causes of death (8 cases) while anaesthetic accidents were responsible for one in six maternal deaths. Some factors were not found such as abruptio placentae, while others such as eclampsia were reduced. We concluded that the maternal mortality rate even though it has been reduced over the last 3 decades it is still high compared with developed countries. We can cut it in half by avoiding maternal transfer in labour.

[Value of ultrasonics in the monitoring of intrauterine devices: apropos of 120 cases]. / Intérêt de l'échographie dans la surveillance des dispositifs intra-utérins: à propos de 120 cas.

PIP: Ultrasound was used in 120 cases to examine women whose IUD strings could not be located during family planning consultations at a clinic in Tunisia. Before the IUD is inserted, ultrasound can be used to determine the exact position of the uterus, identify malformations, locate fibromas, rule out pregnancy, and study the tubes with greater precision. Preliminary ultrasound thus permits identification of contraindications to IUD use. After the IUD is inserted, ultrasound provides more detailed information on the location of the location of the device than either pelvic X-rays or hysterography. The bladder should be full for better analysis of the ultrasound image. 2 kinds of images are created depending on the type of IUD. With copper T devices, the vertical branch is in the form of a continuous, very dense line while the horizontal branch is less dense and may not appear from all angles. The image is similar with a Lippes loop but the dense line is discontinuous. The reported reliability of intrauterine detection of an IUD is almost 100% for all authors. If intraabdominal migration of the IUD has occurred on the long axis of the uterine cavity, the device is easily detected, but if migration was lateral or irregular, the device is more difficult to locate. In 2 cases of intraabdominal migration migration diagnosed by laparoscopy, ultrasound revealed an empty uterus and an image of the device outside the uterine cavity. Ultrasound does not permit diagnosis of common salpingitis, but in all cases of hydro- or pyosalpinx, the image is homogeneous and usually of liquid type, with the IUD visible in the uterus. No other examination can be made in such cases because pelvic infection is an absolute contraindication to all endouterine manipulations. There were 9 cases of pelvic infection in the sample. Ultrasound diagnosis of extrauterine pregnancy is among the most difficult. An empty uterus, a laterouterine image and a liquid image in the pouch of Douglass are highly suggestive of extrauterine pregnancy. 1 case of extrauterine pregnancy occurred; it was suspected on clinical evidence and confirmed by laparoscopy. 6 cases of intrauterine pregnancy were found. When the IUD remains outside the egg, it is usually found almost entirely in the isthmus and cervical canal. Removal is easily accomplished under ultrasound control with no hemorrhagic complications. There were 3 such cases in the series. When the IUD is encrusted in the trophoblast, removal of the device is not recommended unless the patient desires to terminate the pregnancy.^ieng