Dynamic tensile properties of human placenta

Automobile crashes are the largest cause of injury death for pregnant females and the leading cause of traumatic fetal injury mortality in the United States. Computational models, useful tools to evaluate the risk of fetal loss in motor vehicle crashes, are based on a limited number of quasi-static material tests of the placenta. This study presents a total of 20 dynamic uniaxial tensile tests on the maternal side of the placenta and 10 dynamic uniaxial tensile tests on the chorion layer of the placenta. These tests were completed from 6 human placentas to determine material properties at a strain rate of 7.0 strains/s. The results show that the average peak strain at failure for both the maternal portion and the chorion layer of the placenta are similar with a value of 0.56 and 0.61, respectively. However, the average failure stress for the chorion layer, 167.8 kPa, is much higher than the average failure stress for the placenta with the chorionic plate removed, 18.6 kPa. This is due to differences in the structure and function of these layers in the placenta. In summary, dynamic loading data for the placenta have been determined for use in computational modeling of pregnant occupant kinematics in motor vehicle crashes. Moreover the computational model should utilize the material properties for the placenta without the chorion layer.     

Material properties of the placenta under dynamic loading conditions

Trauma during pregnancy especially occurring during car crashes leads to many foetal losses. Numerical modelling is widely used in car occupant safety issue and injury mechanisms analysis and is particularly adapted to the pregnant woman. Material modelling of the gravid uterus tissues is crucial for injury risk evaluation especially for the abruption placentae which is widely assumed as the leading cause of foetal loss. Experimental studies on placenta behaviour in tension are reported in the literature, but none in compression to the authors' knowledge. This lack of data is addressed in this study. To complement the already available experimental literature data on the placenta mechanical behaviour and characterise it in a compression loading condition, 80 indentation tests on fresh placentae are presented. Hyperelastic like mean experimental stress versus strain and corridors are exposed. The results of the experimental placenta indentations compared with the tensile literature results tend to show a quasi-symmetrical behaviour of the tissue. An inverse analysis using simple finite element models has permitted to propose parameters for an Ogden material model for the placenta which exhibits a realistic behaviour in both tension and compression.     

Quantifying dynamic mechanical properties of human placenta tissue using optimization techniques with specimen-specific finite-element models

Motor-vehicle crashes are the leading cause of fetal deaths resulting from maternal trauma in the United States, and placental abruption is the most common cause of these deaths. To minimize this injury, new assessment tools, such as crash-test dummies and computational models of pregnant women, are needed to evaluate vehicle restraint systems with respect to reducing the risk of placental abruption. Developing these models requires accurate material properties for tissues in the pregnant abdomen under dynamic loading conditions that can occur in crashes. A method has been developed for determining dynamic material properties of human soft tissues that combines results from uniaxial tensile tests, specimen-specific finite-element models based on laser scans that accurately capture non-uniform tissue-specimen geometry, and optimization techniques. The current study applies this method to characterizing material properties of placental tissue. For 21 placenta specimens tested at a strain rate of 12/s, the mean failure strain is 0.472±0.097 and the mean failure stress is 34.80±12.62 kPa. A first-order Ogden material model with ground-state shear modulus (μ) of 23.97±5.52 kPa and exponent (α₁) of 3.66±1.90 best fits the test results. The new method provides a nearly 40% error reduction (p<0.001) compared to traditional curve-fitting methods by considering detailed specimen geometry, loading conditions, and dynamic effects from high-speed loading. The proposed method can be applied to determine mechanical properties of other soft biological tissues.     

Effect of frozen storage and aging on the Kashkaval cheese starter culture

Summary The changes in the number of the starter microorganisms Lb. delbrueckii subsp. bulgaricus and Str. thermophiluswere followed in frozen-stored Kashkaval cheese made from cow’s milk. Kashkaval samples of various aging times were produced industrially, frozen at T=−16 °C and stored at T=−10 to −12 °C for 12 months. It was found that the number of Lb. delbrueckiisubsp. bulgaricus and Str. thermophilusdecreased considerably during frozen storage. The decrease was more substantial for Lb. delbrueckiisubsp. bulgaricus, which was evidence for its greater sensitivity to the impact of low temperatures. The aging time of Kashkaval did not influence the changes in the starter culture during frozen storage but is important for its amount in the product aged after defrosting. There was an increase in the Str. thermophilus: Lb. delbrueckiisubsp. bulgaricus ratio in samples with shorter aging time subjected to frozen storage and aged after defrosting. The changes in the starter culture in frozen stored Kashkaval cheese can be controlled by an appropriate combination of the two factors: aging time and period of frozen storage.     

Immunogold localisation of endogenous immunoglobulin-G in ultrathin frozen sections of the human placenta

Summary Endogenous immunoglobulin-G was localised in ultrathin frozen sections of human term placenta by use of an indirect immuno electron-histochemical methodology. Immunoreactivity of endogenous IgG to rabbit anti-human immunoglobulin-G antibody was visualised by use of protein-A — colloidal gold complex. Gold marked the syncytiotrophoblast in both coated and uncoated regions of the apical plasmalemma, in vesicles and multivesicular bodies, and in vesicles near the basal plasmalemma. Immunoreactivity was also seen in the interstitial space between the trophoblast and the fetal endothelial layer as well as in various types of vesicles within the endothelial cells. No immunoreactivity was seen in the intercellular clefts of the endothelium. The pattern of localisation observed is consistent with receptor-mediated uptake of immunoglobulin-G into the syncytiotrophoblast of the human placenta followed by release into the interstitial space and then vesciular transport through the endothelium.     

Purification and properties of arylsulfatase C from human placenta

Arylsulfatase C (ASC) was purified about 1,000-fold from human placenta. The major steps in the procedure included chromatography on Con A-Sepharose and Bio-Gel A-1.5 m. The purified enzyme was homogeneous by sodium dodecylsulfate/polyacrylamide gel electrophoresis. The native enzyme has an apparent molecular weight of 238,000 resulting from three identical subunits of 78,000 daltons. The purified enzyme hydrolyzes the artificial substrate p-nitrophenyl sulfate (NPS), and the two natural substrates estronesulfate (ES) and dehydroepiandrosterone sulfate (DHEAS), the ratio of these three activities being constant throughout the purification. ES and DHEAS are powerful competitive inhibitors of the enzymatic hydrolysis of NPS. ASC, ESase and DHEASase activities show the same thermal stability. These results strongly suggest that a single enzyme is responsible for the hydrolysis of the two natural and the artificial substrates.     

Multipotent properties of myofibroblast cells derived from human placenta

Human uterine fibroblasts (HuF) isolated from the maternal part (decidua parietalis) of a term placenta provide a useful model of in vitro cell differentiation into decidual cells (decidualization, a critical process for successful pregnancy). After isolation, the cells adhere to plastic and have either a small round or spindle-shaped morphology that later changes into a flattened pattern in culture. HuF robustly proliferate in culture until passage 20 and form colonies when plated at low densities. The cells express the mesenchymal cell markers fibronectin, integrin-β1, ICAM-1 (CD54), and collagen I. Flow cytometry of HuF has detected the presence of CD34, a marker of the hematopoietic stem cell lineage, and an absence of CD10, CD11b/Mac, CD14, CD45, and HLA type II. Furthermore, they also express the pluripotency markers SSEA-1, SSEA-4, Oct-4, Stro-1, and TRA-1–81 as detected by confocal microscopy. Treatment for 14–21 days with differentiation-inducing media leads to the differentiation of HuF into osteoblasts, adipocytes, and chondrocytes. The presence of α-smooth muscle actin, calponin, and myosin light-chain kinase in cultured HuF implies their similarity to myofibroblasts. Treatment of the HuF with dimethyl sufoxide causes reversion to the spindle-shaped morphology and a loss of myofibroblast characteristics, suggesting a switch into a less differentiated phenotype. The unique abilities of HuF to exhibit multipotency, even with myofibroblast characteristics, and their ready availability and low maintenance requirements make them an interesting cell model for further exploration as a possible tool for regenerative medicine. Electronic Supplementary Material The online version of this article (doi:) contains supplementary material, which is available to authorized user. This work was supported by National Institutes of Health Grant HD-44713 (to Z.S.).     

Purification and various properties of uracil-DNA-glycosylase from human placenta

Uracil-DNA-glycosylase was isolated from human placenta and purified 2100-fold. The apparent Km value for non-methylated DNA substrate of the enzyme is 3.10(-7) M. However, Km for uracil-DNA-glycosylase was 3 times as low when methylated DNA was used as a substrate. It was shown that the initial rate of uracil excision was greater for the non-methylated than for the hypermethylated DNA. The experimental results indicate that the postreplicative methylation of DNA can interfere with uracil excision.     

Purification and properties of nucleotide pyrophosphatase from human placenta

Nucleotide pyrophosphatase was purified from human placenta to near homogeneity with a specific activity of about 500-fold over the Triton extract of the homogenate. Purification was achieved most effectively by successive chromatographic steps with AMP-agarose and ADP-agarose columns, based on the affinity of the enzyme towards 5'-adenylate and adenosine 3',5'-diphosphate, and a lectin-Sepharose column, based on the glycoprotein nature of the enzyme. The purified enzyme was found to be essentially homogeneous on SDS-polyacrylamide gel electrophoresis with a mobility corresponding to 130K. The purified enzyme was found to hydrolyze a wide variety of nucleotides, i.e. 3'-phosphoadenosine 5'-phosphosulfate (PAPS), adenosine 5'-phosphosulfate (APS), NADH, ATP, nucleotide sugars, oligonucleotides, and p-nitrophenyl-thymidine 5'-phosphate (PNTP). From the oligonucleotides, the enzyme produced 5'-phosphates. Mg2+ was required for full activity. Glycine and sulfhydryl compounds such as 2-mercaptoethanol and 2,3-dimercapto-1-propanol were inhibitory. Most of these properties are common to nucleotide pyrophosphatases [EC 3.6.1.9] and type I (5'-phosphate forming) phosphodiesterases [EC 3.1.4.1] from various sources. The relevance of this enzyme to a unique genetic disease, Lowe's syndrome, is discussed.     

Purification and kinetic properties of galactokinase from human placenta

Galactokinase from human placenta was purified about 350-fold using DEAE-Sephadex-A-50 chromatography followed by Sephadex-G-200 and CM-Sephadex-C-50 filtration. The final steps of purification involved electrofocusing and ammonium sulfate precipitation. In analytical disc electrophoresis the purified enzyme moved as a single protein band.     

Purification and properties of human placenta arylsulphatase A.

1. Arylsulphatase A (arylsulphate sulphohydrolase E.N. 3.1.6.1) has been purified 7200-fold from human placenta using concanavalin A Sepharose chromatography. 2. Ultracentrifugation studies indicated that the purified enzyme was homogenous with respect to sedimentation coefficient and molecular weight and has a molecular weight of 102000. 3. The purified enzyme could hydrolyze cerebroside 3-sulphate, seminolipid and sulphogalactosylsphingosine under identical conditions. 4. The kinetic parameters for the hydrolysis of all sulphate esters used in the present study were the same. 5. Both seminolipid and sulphogalactosylsphingosine were competitive inhibitors for the hydrolysis of cerebroside-3-sulphate with an inhibition constant of 0.2 mM. 6. Kinetic parameters, metal ion effect and heat inactivation profile of enzyme suggest that the same active site of enzyme is responsible for the hydrolysis of cerebroside 3-sulphate, seminolipid and sulphogalactosylsphingosine.     

Purification and properties of aldehyde reductases from human placenta

Aldehyde reductases (alcohol: NADP⁺-oxidoreductases, EC 1.1.1.2) I and II from human placenta have been purified to homogeneity. Aldehyde reductase I, molecular weight about 74 000, is a dimer of two nonidentical subunits of molecular weigths of about 32 500 and 39 000, whereas aldehyde erductase II is a monomer of about 32 500. Aldehyde reductase I can be dissociated into subunits under high ionic concentrations. The isoelectric pH for aldehyde reductases I and II are 5.76 and 5.20, respectively. Amino acid compositions of the two enzymes are significantly different. Placenta aldehyde reductase I can utilize glucose with a lower affinity, whereas aldehyde reductase II is not capable to reducing aldo-sugars. Similarly, aldehyde reductase I does not catalyse the reduction of glucuronate while aldehyde reductase II has a high affinity for glucuronate. Both enzymes, however, exhibit strong affinity towards various other aldehydes such as glyceraldehyde, propionaldehyde, and pyridine-3-aldehyde. The pH optima for aldehyde reductases I and II are 6.0 and 7.0, respectively. Aldehyde reductaase I can use both NADH and NADPH as cofactors, whereas aldehyde reductase II activity is dependent on NADPH only. Both enzymes are susceptible to inhibition by sulfhydryl group reagents, aldose reductase inhibitors, lithium sulfate, and sodium chloride to varying degrees.     

Effect of hydration on the molecular dynamic properties of human serum albumin at low temperatures

The effect of temperature and hydration on phosphorescence of chromatophores and on saturation curves of ESR spectra of spin labels covalently bound to human serum albumin was studied. It has been shown that at 90-260 degrees K albumin hydration results in intensification of motions of hydrophobic parts with low frequencies (vc less than or equal to 10(3) s-1) and does not affect the motions of hydrophobic and surfacial parts with high frequency.