BackgroundMesenchymal stromal cells are multipotent cells considered to be of great promise for use in regenerative medicine. However, the cell dose may be a critical factor in many clinical conditions and the yield resulting from the ex vivo expansion of mesenchymal stromal cells derived from bone marrow may be insufficient. Thus, alternative sources of mesenchymal stromal cells need to be explored. In this study, mesenchymal stromal cells were successfully isolated from second trimester amniotic fluid and analyzed for chromosomal stability to validate their safety for potential utilization as a cell therapy product.

In utero diagnosis of ultrasound patterns associated with DV anomalies is feasible. Fetal karyotyping should be considered, serial ultrasound examinations recommended and, in the presence of heart failure, delivery can be anticipated.

To determine whether S100␤, an acidic calcium-binding protein previously demonstrated as a reliable indicator of a brain lesion, could be helpful in the detection of brain distress in intrauterine growth-retarded (IUGR) fetuses, we studied, by a case-control study, the correlation between S100B protein and the degree of fetoplacental blood flow impairment. Maternal and umbilical blood samples and placental tissue specimens were collected at delivery from IUGR pregnancies with normal (n ϭ 10) or abnormal (n ϭ 10) umbilical artery Doppler findings and from 40 uncomplicated pregnancies. S100␤ protein levels were measured by means of a specific RIA, and flow velocimetry waveforms were recorded from uterine, umbilical, and fetal middle cerebral arteries. Overall mean S100␤ proteins in umbilical plasma levels were higher (p Ͻ 0.05) in IUGR patients (121.8 Ϯ 70.4 fmol/mL) than in control patients (54.7 Ϯ 21.9 fmol/mL). IUGR fetuses with redistribution of blood flow showed the higher concentration of the protein (163.7 Ϯ 55.2 fmol/mL).Fetal S100␤ concentrations correlated with middle cerebral artery pulsatility index (r ϭ Ϫ0.536, p Ͻ 0.03) and with umbilical artery pulsatility index to middle cerebral artery pulsatility index ratio (r ϭ 0.469, p Ͻ 0.03). No difference in the localization or intensity of S100␤ staining in the placental tissues or cord between uncomplicated and IUGR pregnancies was found. This study provides evidence that circulating S100␤ protein is increased in IUGR fetuses and correlates with cerebral hemodynamics, suggesting that it may represent an index of cerebral cell damage in the perinatal period. The S100 family of calcium-binding proteins, first isolated in 1965 by Moore (1) in a subcellular fraction from bovine brain, contains approximately 16 members, each of which exhibits a unique pattern of tissue-or cell type-specific expression. Although the distribution of these proteins is not restricted to the nervous system, the involvement of several members of this family in nervous system development, function, and disease has sparked new interest in these proteins. S100␤, one of the original two members of this family, is an acidic calcium-binding protein with a molecular weight of 21 kD. It is present extracellularly, intracellularly, and in the cytosol; its half-life is approximately 2 h, and it is mainly eliminated by the kidney (2). S100␤ is present in CNS and is concentrated in the glial cells, astrocytes, Schwann cells, and neurons. It regulates several cellular functions (cell-cell communication, cell growth, cell structure, energy metabolism, contraction, and intracellular signal transduction). Elevated plasma levels are found in patients with brain damage (3). Abnormal S100␤ levels have been associated with neurobehavioral abnormalities and microcephaly caused by in utero cocaine exposure (4), and abnormal S100␤ immunoreactivity cells in anencephalic fetuses have been shown (5).The S100␤ concentration in blood and in cerebrospinal fluid is increased as result of brain damage in adults a...

nisolone produced a weak signal at a slightly different retention time. At the evaluated concentration of 5000 g/L, prednisolone produced a signal equivalent to 20 g/L cortisol, representing a 0.4% interference. Fenofibrate generated parent ions at m/z 361 and 363 in an abundance ratio of 3:1, a distinctive pattern related to the presence of chlorine in the chemical composition. In MS/MS mode, fenofibrate produced a m/z 3633121 transition that interfered with the quantitative transition of cortisol, but it did not produce the m/z 363397 transition. Thus, switching to the secondary transition for quantification eliminated interference from this drug. In addition, the elution time of the drug was ϳ30 s longer than that of cortisol.Taylor et al.(1 ) noted isotopic exchange between the deuterated IS and hydrogen-containing vapors in an APCI ion source. Unlike the observations of Taylor et al., we have not observed isotopic exchange with our APCI interface. The reason for this difference is unclear, but it may be related to ion source conditions. The good agreement obtained between our method and the comparison LC-MS/MS method suggests that isotopic exchange did not affect the proposed method.Apparently healthy adult volunteers (25 males and 25 females; age range, 19 -53 years) collected 24-h urine samples without preservative. The volunteers were asked to keep samples under refrigeration during the collection. Statistical results for this study are presented in Table 1.To evaluate agreement between the established reference interval with UFC values in the population, we evaluated results for 2089 random 24-h urine specimens analyzed with the proposed method (Fig. 1). The mean (SD) value for log-transformed UFC excretion in 24 h was 1.26 (0.24) log g/24 h (minimum and maximum, 0.3 and 3.84 log g/24 h, respectively), and skewness and kurtosis for the distribution were 1.51 and 5.57, respectively.The stability of cortisol in urine was evaluated in the presence of acetic (15 mmol/L), boric (15 mmol/L), and hydrochloric (30 mmol/L) acid. Two samples without added acid were stored and analyzed under the same conditions as the samples stored with the acids. Samples were stored at room temperature, 4°C, and Ϫ20°C and analyzed every 4 -7 days during 1 month of storage. Cortisol concentrations in samples stored with the acids were higher by ϳ30% than in samples stored without acid, possibly as a result of partial hydrolysis of sulfate and glucuronide conjugates.In conclusion, the rapid LC-MS/MS method for UFC analysis appears to be free from interference and agrees closely with a HPLC-MS/MS method that uses sample extraction. The method has been demonstrated reliable in a high-volume clinical laboratory environment.

Longitudinal S100B protein measurements in urine soon after birth are a useful tool to identify which asphyxiated infants are at risk of long-term neurologic sequelae.