Overexpression of SMN2 Gene in Motoneuron-Like Cells Differentiated from Adipose-Derived Mesenchymal Stem Cells by Ponasterone A.

Cell therapy and stem cell transplantation strategies have provided potential therapeutic approaches for the treatment of neurological disorders. Adipose-derived mesenchymal stem cells (ADMSCs) are abundant adult stem cells with low immunogenicity, which can be used for allogeneic cell replacement therapies. Differentiation of ADMSCs into acetylcholine-secreting motoneurons (MNs) is a promising treatment for MN diseases, such as spinal muscular atrophy (SMA), which is associated with the level of SMN1 gene expression. The SMN2 gene plays an important role in MN disorders, as it can somewhat compensate for the lack of SMN1 expression in SMA patients. Although the differentiation potential of ADMSCs into MNs has been previously established, overexpression of SMN2 gene in a shorter period with a longer survival has yet to be elucidated. Ponasterone A (PNA), an ecdysteroid hormone activating the PI3K/Akt pathway, was studied as a new steroid to promote SMN2 overexpression in MNs differentiated from ADMSCs. After induction with retinoic acid, sonic hedgehog, forskolin, and PNA, MN phenotypes were differentiated from ADMSCs, and immunochemical staining, specific for ß-tubulin, neuron-specific enolase, and choline acetyltransferase, was performed. Also, the results of real-time PCR assay indicated nestin, Pax6, Nkx2.2, Hb9, Olig2, and SMN2 expression in the differentiated cells. After 2 weeks of treatment, cultures supplemented with PNA showed a longer survival and a 1.2-fold increase in the expression of SMN2 (an overall 5.6-fold increase; *P ≤ 0.05), as confirmed by the Western blot analysis. The PNA treatment increased the levels of ChAT, Isl1, Hb9, and Nkx2 expression in MN-like cells. Our findings highlight the role of PNA in the upregulation of SMN2 genes from MSC-derived MN-like cells, which may serve as a potential candidate in cellular therapy for SMA patients.

Early infantile presentation of 3-methylglutaconic aciduria type 1 with a novel mutation in AUH gene: A case report and literature review.

3-Methylglutaconic aciduria is a member of inborn errors of leucine metabolism pathway. 3-Methylglutaconic aciduria type I (MGA1) causes neurological problems which are present during infancy or childhood but the diagnosis may be delayed until adulthood. Here we report a 3years old patient with developmental delay from a relative parent's that his medical evaluations include analyses of urinary organic acid and blood acylcarnitine showed high level of 3-methylglutacoic acid, 3-hydroxyisovaleric acid and increased level of 3-hydroxyisovalerylcarnitine respectively. Further evaluation and genetic tests revealed a novel homozygous mutation of variant c.179del G (p.Gly60Valfs*12) in exon 1 of the AUH gene that was compatible with the diagnosis of MGA1. In segregation analysis of his family, both parents were heterozygous for the respective mutation, confirming obligate parental carrier status and segregation of the mutation.

Sudden impolite behavior in a polite girl: a case report.

In this case we present a previously healthy child with sudden behavioral change and acute confusional state (ACS). ACS is a neurologic emergency. The clinical manifestations of ACS are; confused affect, limited verbal response, disturbance in performing orders, some automatism, stable vital signs and absence of tonic-clonic movements and it has a wide range of differential diagnosis which should be assessed by detailed medical history, medical examination and laboratory tests. The only abnormal test found in this patient, was cerebrospinal fluid abnormality, consistent with viral encephalitis.

Impact of nurses clothing on anxiety of hospitalised children.

AIM: To investigate anxiety levels in two groups of children exposed to nurses with white vs. coloured clothing in a university hospital in Iran. BACKGROUND: Hospitalisation causes anxiety in children and it is documented that nurses have an important role in alleviating children's distress and anxiety. Nurses characteristics, including their clothing is a factor that affects quality of care through child-nurse relationship. DESIGN: Clinical trial. METHODS: Children (n = 92) aged 7-15 years old hospitalised for 3-5 days in paediatric surgery ward were exposed to nurses in white or coloured clothing. Children's anxiety was assessed on admission and at discharge using Revised Children's Manifest Anxiety Scale. RESULTS: Children exposed to white nursing uniforms showed higher anxiety levels compared with children exposed to coloured nursing clothing (p < or = 0.05). Besides coloured nursing clothing, female sex, age >11 years old (guidance school) and living in families with more than four members were predictors of lower global anxiety scores. CONCLUSION: Providing a child-friendly environment through colourful nursing clothing can promote nurses' relationship with hospitalised children. This can satisfy children's expectations of the nursing care and alleviates the need for meeting ideals of nursing care through wearing a white nursing uniform provided that standards of nursing care are favoured. RELEVANCE TO CLINICAL PRACTICE: Using colourful nursing clothing in paediatric wards reduces anxiety as a psychological parameter which delays improvement and provides a child-friendly environment that helps promotion of quality of nursing care.

The effects of amniotic fluid on the histopathologic changes of exposed spinal cord in fetal sheep.

BACKGROUND: Experimental studies have shown that in myelomeningocele, the primary malformation is neural tissue damage resulting from exposure of neural tissue to amniotic fluid. In this study, the effects of amniotic fluid on histopathologic changes of exposed spinal cord in fetal sheep were evaluated. METHODS: In an experimental trial, 10 fetal sheep in two groups containing five subjects (group A) and five shams (group B) were studied. In the sheep at 90 - 100 days of gestation (term: 145 - 150 days) the lumbar skin was incised, paraspinal soft tissues were excised, laminectomy was performed at L2 - L4, and dura matter was opened. In group A, the dura matter was not dorsally closed and thus the spinal cord was left exposed to amniotic fluid, and in group B the skin was immediately closed. The lambs were delivered near term by cesarean section and were assessed clinically and morphologically. RESULTS: In group A, all lambs (n=5) had a complete or incomplete flaccid sensorimotor paraplegia and suffered from urine incontinence. Four lambs in this group were stool incontinent. In group B (n=4), only one lamb had paraparesis (P=0.048) and all lambs were urine and stool continent. In group A, all lambs had hypoplastic longitudinal muscles of the rectum but well- developed circular muscles. The anal sphincter muscles did not develop normally. In group B, all lambs had well-developed longitudinal and circular muscles and anal sphincter muscles developed normally (P=0.048). Histopathologic examination of the spinal cords showed edema, focal calcification, fibrosis, and capillary cell proliferation in group A, but in group B such changes were not seen. The number of ganglion cells was significantly higher in group B compared with group A (P<0.0005). CONCLUSION: Exposure of spinal cord to amniotic fluid causes structural neural tissue damage that can be prevented by fetal surgery through repairs of myelomeningocele.

Evaluation of diluted amniotic fluid effects on histological changes of intestine of rabbit fetus with gastroschisis.

Amniotic fluid exchange is a method for prevention of intestinal damage in gastroschisis, but its techniques are different in studies. We investigated the effects of amnioinfusion exchange on histological changes of intestine and feasibility and safety of amniotic fluid exchange through central vein catheter (CVC) placed in pregnant rabbit uterus. A total of 15 pregnant New Zealand white rabbits were selected. Fetuses were randomly divided into three groups (case, control, sham). On gestational day 25, under general anesthesia with midline laparotomy, the graved bicornuate uterus was exposed. In controls, fetus abdomen was opened by a transverse incision in right lower quadrant region and intestines were eviscerated. In cases, after intestine evisceration, a central venous catheter was passed from mother skin and uterus and fixed to uterus wall. In shams, fetus was delivered on gestational day 32 and its abdomen was opened. In case group, after operation, 1-2 cc of warm saline solution was replaced through catheter every 6 h. On gestational day 32, fetuses of case and control groups were delivered. Mucosal and serosal thickness, muscle thickness, fibrin deposition, serosal collagen and ganglia were compared. Ten fetuses as shams, 7 fetuses as controls and 7 fetuses as case group were studied. Serosal thickness was 4.5 +/- 3.6 microm in shams, 64.2 +/- 28.7 microm in controls and 6 +/- 4.1 microm in cases. Serosal thickness in control group was higher than sham (P < 0.001) and case (P < 0.002) groups. In case group, infiltration of inflammatory cells with mild edema without fibroblast infiltration was seen. Application of the CVC technique was found to be a simple procedure that effectively decreased serosal inflammatory response of intestine in gastroschisis.

Selenium and intractable epilepsy: is there any correlation?

Oxidative stress and generation of reactive oxygen species are strongly implicated in a number of neuronal and neuromuscular disorders, including epilepsy. The functions of selenium as an antioxidant trace element are believed to be carried out by selenoproteins that possess antioxidant activities and the ability to promote neuronal cell survival. Because of this protective role of selenium against oxidative damage, a case-control study was designed to compare its serum level between intractable epileptic patients and normal subjects. Eighty patients who met the criteria of intractable epilepsy were compared with a normal control group of the same age, socioeconomic level, and place of living. Serum selenium level was measured with an atomic absorption spectrophotometer. The mean (+/- S.D.) of serum selenium were 68.88 (+/-17.58) ng/mL and 85.93 (+/-13.93) ng/mL in the patient and control groups respectively. Independent sample t test with P < 0.05 indicated a significant lower mean of serum selenium in the patient group compared with that of the normal control group. However, there was no association between serum selenium and some suggested predictive factors of intractable seizures, including age at the onset of seizures, neonatal seizure, neurologic impairment, and etiology of epilepsy. Measurement of serum selenium in patients with intractable epilepsy should be considered.

Extensive Mongolian spots: a clinical sign merits special attention.

Although typical and limited Mongolian spots are benign skin markings at birth which fade and disappear as the child grows, extensive Mongolian spots deserve special attention as possible indications of associated inborn error of metabolism. A few cases of extensive Mongolian spots in association with inheritable storage diseases have been reported. Some hypotheses have been put forward, but further investigation is necessary to elucidate the causative factors. This report describes three infants with generalized Mongolian spots, two infants with GM1 gangliosidosis type 1, and one in association with Hurler syndrome. Findings of generalized Mongolian spots in newborns may lead to an early detection and early treatment before irreversible organ damage occurs.

Pancreatitis after liver transplantation in children: a single-center experience.

BACKGROUND AND METHOD: Posttransplantation acute pancreatitis (PTAP) is a rare but serious complication after pediatric liver transplantation (LTx). We performed a retrospective review in a large cohort of pediatric liver transplant recipients at a single institution to define the impact of this problem in children. RESULTS: Between January 1986 and December 1999, 634 pediatric LTx were performed. Twenty-six patients developed serious acute pancreatitis. The mean age at transplantation was 7.7 years (9 months to 19 years), and the indications for transplantation were biliary atresia in seven, fulminant hepatic failure in six, chronic rejection in seven, and other etiologies in six patients. PTAP was more likely to occur early after LTx (61% within the first week), was associated with the presence of an infrarenal aortic graft in 14 (54%) of 26 patients, was more likely to occur after retransplantation (11/26 patients), and was associated with blood loss and prolonged surgery in four cases. Acute renal failure occurred in 15 (58%) of 26 patients. Mortality was 42% (11/26); causes of death were sepsis or multiple organ failure in nine and hemorrhage in two patients. Management of PTAP included antibiotics, sphincterotomy, debridement with drainage, hepatic arterial revascularization, and arterial ligation. Of the 14 patients with complicated pancreatitis, 5 were treated conservatively and died. Nine patients had extensive operative interventions and four survived (45%). CONCLUSIONS: Several risk factors such as retransplantation, extensive dissection at the time of LTx, and use of infrarenal arterial graft contribute to development of PTAP in children. Early exploration and debridement in patients with complicated pancreatitis may result in a better outcome. Retransplantation in the presence of clinical pancreatitis has a high failure rate.