Özlem Giray

Erythropoietin attenuates lipopolysaccharide-induced white matter injury in the neonatal rat brain.

Periventricular leukomalacia (PVL), a common neonatal brain white matter (WM) lesion, is frequently associated with cerebral palsy. Growing evidence has indicated that in addition to ischemia/reperfusion injury, cytokine-induced brain injury associated with maternal or fetal infection may also play an important role in the pathogenesis of PVL. Recent studies have shown that administration of lipopolysaccharide (LPS) to pregnant rats causes enhanced expression of the cytokines, i.e., IL-1β, TNF-α, and IL-6, in fetal brains. In recent years, it has been shown that erythropoietin (EPO) has a critical role in the development, maintenance, protection and repair of the nervous system. In the present study we investigated the effect of EPO on LPS-induced WM injury in Sprague-Dawley rats. LPS (500 µg/kg) suspension in pyrogen-free saline was administered intraperitoneally to pregnant rats at 18 and 19 days of gestation. The control group was treated with pyrogen-free saline. They were given 5,000 U/kg recombinant human EPO. Seven-day-old Sprague-Dawley rat pups were divided into four groups: control group, LPS-treated group, prenatal maternal EPO-treated group (5,000 U/kg, intraperitoneally given to pregnant rats at 18 and 19 days of gestation), and postnatal EPO-treated group (5,000 U/kg, intraperitoneally given to 1-day-old rat pups). Cytokine induction in the postnatal 7-day-old (P7) rat brain after maternal administration of LPS was determined by the ELISA method. The proinflammatory cytokine levels (IL-1β, TNF-α, and IL-6) in P7 rat pup brains were significantly increased in the LPS-treated group as compared with the control group. Prenatal maternal EPO treatment significantly reduced the concentration of TNF-α and IL-6 in the newborn rat brain following LPS injection. The concentration of IL-1β was decreased in the intrauterine EPO treatment group. Postnatal EPO treatment significantly decreased only the IL-6 concentration in the newborn rat brain following LPS injection. The concentration of cytokines, IL-1β and TNF-α, was reduced in the postnatal EPO treatment group. We demonstrated here that LPS administration in pregnant rats at gestational day 18 and 19 induced WM injury in P7 progeny characterized by apoptosis. Prenatal maternal and postnatal EPO treatment significantly reduced the number of apoptotic cells in the periventricular WM. Using immunohistochemistry techniques, we investigated the effects of maternal administration of LPS on myelin basic protein (MBP) staining, as a marker of myelination in the periventricular area in the neonatal rat brain. MBP staining was significantly less and weaker in the brains of the LPS-treated group as compared with the prenatal maternal EPO-treated group. However, the postnatal EPO treatment did not prevent LPS-stimulated loss of MBP-positive staining. In conclusion, especially prenatal maternal EPO treatment attenuates LPS-induced injury by reducing the expression of inflammatory cytokines and sparing MBP in the neonatal rat brain. While the postnatal EPO treatment prevented LPS-induced brain injury this effect was partial. To our knowledge, this is the first study that demonstrates a protective effect of EPO on LPS-induced WM injury in the developing brain. Regarding the wide use of EPO in premature newborns, this agent may be potentially beneficial in treating LPS-induced brain injury in the perinatal period.

Activated protein C reduces endotoxin-induced white matter injury in the developing rat brain.

Periventricular leukomalacia (PVL), the dominant form of brain injury in premature infants, is characterized by white matter injury (WMI) and is associated with cerebral palsy. The pathogenesis of PVL is complex and likely involves ischemia/reperfusion, free radical formation, excitotoxicity, impaired regulation of cerebral blood flow, a procoagulant state, and inflammatory mechanisms associated with maternal and/or fetal infection. Using an established animal model of human PVL, we investigated whether activated protein C (APC), an anti-coagulant factor with anti-inflammatory, anti-apoptotic, anti-oxidant, and cytoprotective activities, could reduce endotoxin-induced WMI in the developing rat brain. Intraperitoneal injections of lipopolysaccharide (LPS) (0.5 mg/kg body weight) were given at embryonic days 18 (E18) and 19 (E19) to pregnant Sprague-Dawley rats; control rats were injected with sterile saline. A single intravenous injection of recombinant human (rh) APC (0.2 mg /kg body weight) was given to pregnant rats following the second LPS dose on embryonic day 19 (E19). Reduced cell death in white matter and hypomyelination were shown on TUNEL and myelin basic protein (MBP) staining, respectively, on late postnatal days (P7) in APC-treated groups. There were significantly fewer TUNEL+nuclei in the periventricular WM in the APC+LPS group than in the untreated LPS group. Compared to the APC+LPS and control group, MBP expression was weak in the LPS group on P7, indicating endotoxin-induced hypomyelination in the developing rat brain. APC attenuated the LPS-induced protein expression of inflammatory cytokines, tumor necrosis factor-alpha, and interleukin-6, as evaluated by ELISA in neonatal rat brains. A single intraperitoneal injection of rhAPC (0.2 mg/kg body weight) to neonatal rats on P1 also had similar protective and anti-inflammatory effects against maternally administered LPS. Collectively, these data support the hypothesis that APC may provide protection against an endotoxin-evoked inflammatory response and WMI in the developing rat brain. Moreover, our results suggest that the possible use of APC in treatment of preterm infants and pregnant women with maternal or placental infection may minimize the risk of PVL and cerebral palsy.

Analyses of Polymorphism for UGT1*1 Exon 1 Promoter in Neonates with Pathologic and Prolonged Jaundice

In this study, we investigated whether a TATA box polymorphism in the promoter of the UGT1*1 exon I, the most common detected DNA polymorphism in Gilbert’s syndrome, is a contributory factor in unexplained pathologic or prolonged jaundice. 38 neonates who had unexplained pathologic jaundice, 37 neonates who had unexplained prolonged jaundice, and 35 healthy, nonjaundiced neonates were enrolled in the study. Genotypes were assigned as follows: 6/6 (homozygous for a normal allele bearing the sequence [TA]6TAA), 7/7 (homozygous for an abnormal allele with the sequence [TA]7TAA), and 6/7 (heterozygous with one of each allele). Of the 110 infants, 10 (9%) had 7/7, 51 (46%) had 6/7, and 49 (45%) had 6/6 genotype; the differences between the three groups were not statistically significant. Also no differences were observed among different genotypes and mean serum total bilirubin concentrations. In conclusion, we showed that TA 7/7 and TA 6/7 genotypes are not rare in our population and that the presence of these polymorphisms alone does not play a significant role in the etiology of unexplained pathologic or prolonged neonatal hyperbilirubinemia.

Clinical and morphological phenotype of geleophysic dysplasia

Abstract Geleophysic dysplasia (GD) is a rare, recessively inherited lysosomal storage disorder of unknown origin with a progressive course. A 9-year-old Turkish boy born to consanguineous parents with findings typical of GD is reported. Cardiac abnormalities included mitral and aortic stenosis with aortic insufficiency. There was persistent hypo-uricacidaemia, severe pulmonary hypertension and tricuspid insufficiency. He required aortic and mitral valve replacement but, unfortunately, died of a severe pulmonary infection in the post-operative period. The condition has to be differentiated from lysosomal storage disorders such as mucopolysaccharidosis.

Long-standing fever and Angelman syndrome: report of two cases.

Abstract:  An 8‐month‐old girl and a 20‐month‐old boy who presented with motor and developmental delay and long‐standing fever are presented. The patients were diagnosed as Angelman syndrome with fluorescence in situ hybridization (FISH) analysis. Despite extensive clinical and laboratory examinations, no inflammatory or infectious origin for the fever was found. It was considered that the long‐standing fever observed in these cases was due to hypothalamic dysfunction for thermoregulation.

Role of Apolipoprotein E in Febrile Convulsion

Apolipoprotein E is consistently associated with the progression of some common human neurodegenerative diseases, e.g., epilepsy. We hypothesized that genetic variations in the apolipoprotein E gene have implications for susceptibility to, and prognoses in, febrile convulsion, which plays an apparent role in the development of epilepsy. We used the polymerase chain reaction and restriction enzyme digestion to characterize variations of the apolipoprotein E gene. Sixty-nine patients with febrile convulsion (simple/complex) and a corresponding cohort of healthy patients (n = 75) were used. There was no significant difference in genotypic distribution and allelic frequencies of the apolipoprotein E gene between the febrile convulsion and control groups. Comparing subpopulations of the febrile convulsion group (patients with simple and complex febrile convulsion), we noted that no patients with the epsilon3/epsilon4 genotype had complex febrile convulsions. The apolipoprotein E epsilon3/epsilon4 genotype was more frequently seen in the simple febrile than in the complicated febrile convulsion group (9 versus 0 patients, respectively). The data indicate an association with the epsilon3/epsilon4 genotype of the apolipoprotein E gene with a milder phenotype. Although apolipoprotein E4 is not a vulnerability factor regarding febrile convulsions, it seems effective in regard to prognoses.

Multi-cystic white matter enlarged Virchow Robin spaces in a 5-year-old boy

Virchow–Robin spaces (VRS) are perivascular, pial-lined areas in the brain that surrounds small arteries and can be seen on brain magnetic resonance imaging (MRI) [6, 9]. On MRI sequences, VRS are fluid-filled spaces, and their signal intensities are identical to those of cerebrospinal fluid [4, 22]. VRS are typically less than 5 mm in diameter and accepted as normal anatomical structures. The VRS around the perforating arteries are in continuity with the perivascular spaces around arterioles in the subpial region, and when they perforate the brain surface and extend into the brain tissue, they are usually called enlarged VRS [3, 7, 21, 24]. Dilatation of VRS may be present in physiological conditions and many different diseases such as mucopolysaccharidosis, mannosidosis, Lowe syndrome, Coffin–Lowry syndrome, and ectodermal dysplasia [9–11, 15, 17]. In childhood, dilatation of VRS is rarely secondary to specific progressive disorders but usually appears in developmental delay [4]. VRS are usually asymptomatic, but if there is an expansion to the brain parenchyma, various clinical symptoms can be seen associated with the mass effect [21]. Extreme dilatation of VRS has been reported with different clinical findings, including headache, tics, dementia, seizures, extrapyramidal symptoms, ataxia, developmental delay, macrocephaly, or no neurologic abnormalities [1, 2, 5]. The visibility of VRS is dependent on the MRI sequence that is used [9]. T2-weighted MRI sequences lead to better visualization of VRS [20]. On MRI T2-weighted imagings, VRS are isointense relative to CSF and show no enhancement after contrast agent administration [21]. T1-weighted images show hyposignal intensity [4]. By the radiological appearances, that can be differentiated from cystic neoplasia, parasitic infections, cystic infarction, and some metabolic disorders [6, 10, 13]. VRS can occur in many different locations, and the most common location is around the lenticulostriate arteries. Occasionally, that occurs around the arteries which have perforated the cerebral cortex and extend into the white matter [10, 13, 21]. In this report, we described the 2-years follow-up of a 5-year-old boy with enlarged VRS, developmental delay, macrocephaly, hair with a double crown, and clinodactyly. The patient was not consistent with known syndrome descriptions. The symptoms of this patient may belong to a new syndrome that was previously unidentified.

Maternal preeclampsia and jitteriness in preterm infants.

Abstract Aim: To clarify the role of maternal preeclampsia in jitteriness in preterm infants.

Multiple pterygium syndrome with horseshoe kidney and polydactyly: a further case.

Case report The patient, a 7-year-old male born to nonconsanguineous parents was diagnosed to have arthrogryposis multiplex congenital at birth. His weight was 2510 g, length 42 cm, without correction for the severe contractures and head circumference 35 cm. A previous stillborn sibling had also had multiple webs involving the joints. In the newborn period, the patient was noted to have generalized hirsutism, unilateral postaxial polydactyly of the left hand, bilateral talipes equinovarus deformity, flexion deformity at the wrists, and cryptorchid testes. Abdominal ultrasound examination showed a horseshoe kidney. Development was normal and results of his cognitive evaluation revealed that he was performing within the average range of intellectual functioning. He was diagnosed to have multiple pterygium syndrome (MPS) based on his clinical findings (Fig. 1a and b).