Concentrations Of Fetal Serum Research Articles

Primary Human Fibroblasts in Culture Switch to a Myofibroblast-Like Phenotype Independently of TGF Beta.

Fibroblasts are the prevalent cell type and main source for extracellular matrix (ECM) in connective tissue. Depending on their origin, fibroblasts play a central role in non-pathological tissue remodeling and disease like fibrosis. This study examined the effect of established culture conditions of primary human fibroblasts, from different origins on the myofibroblast-like phenotype formation. We isolated primary human fibroblasts from aortic adventitia, lung, juvenile- and adult skin and investigated the expression levels of CD90, alpha smooth muscle actin (αSMA) and procollagen I under different concentrations of fetal calf serum (FCS) and ascorbic acid (AA) in culture media by immunoblot and immunofluorescence assays. Furthermore, we determined the viability using XTT and migration/wound healing in scratch assays. Collagen 1 secretion was quantified by specific ELISA. Primary human fibroblasts show in part a myofibroblast-like phenotype even without addition of FCS. Supplemented AA reduces migration of cultured fibroblasts with no or low concentrations of FCS. Furthermore, AA and higher concentrations of FCS in culture media lead to higher levels of collagen 1 secretion instead of procollagen I accumulation. This study provides evidence for a partial switch of primary human fibroblasts of different origin to a myofibroblast-like phenotype under common culture conditions.

Isolation and culture of pleural mesothelial cells

Purpose: Although the isolation of rat and mouse mesothelial cells has previously been reported, most mesothelial cells used for experimental studies are obtained from peritoneal cells. Here, we describe an optimized method for the isolation and in vitro propagation of rodent pleural mesothelial cells without the requirement for specialized surgical techniques. Materials and Methods: To harvest pleural mesothelial cells, the pleural space of 8–9-week-old rats or older mice was filled with 0.25% trypsin in ethylenediaminetetraacetic acid (EDTA) buffer for 20 min at 37 °C. Cells were then harvested, and incubated at 37 °C in a humidified atmosphere with 5% CO2. Immunofluorescence analysis of plated pleural mesothelial cells was performed using Alexa 546 (calretinin). To investigate optimal proliferation conditions, medium enriched with various concentrations of fetal calf serum (FCS) was used for pleural mesothelial cell proliferation. Results: By day 10, confluent cell cultures were established, and the cells displayed an obvious cobblestone morphology. Immunofluorescence analysis of the cells demonstrated that all stained positive for Alexa 546 (calretinin) expression. Mesothelial cells grew better in medium containing 20% FCS than with 10% FCS. Conclusions: This is a simple procedure for the efficient collection of primary pleural mesothelial cells, which were obtained in defined culture conditions from the euthanized rodent thoracic cavity using trypsin-EDTA treatment. The ability to easily culture and maintain identifiable pleural mesothelial cells from rodents will be helpful for future experiments using these cells.

Impacto de la concentración de suero fetal bovino sobre las células epiteliales dentales

El esmalte dental, tejido más duro del cuerpo humano, es formado por medio de la diferenciación de células epiteliales conocidas como ameloblastos. Recientemente, las células epiteliales dentales de incisivo de rata han sido utilizadas como modelo celular de eventos fisiopatológicos durante la formación del esmalte dental. Sin embargo, en función de los eventos estudiados es meritorio comprender su comportamiento, particularmente cuando la concentración del Suero Fetal Bovino (SFB) varia. El propósito de este trabajo es evaluar el impacto de la concentración del SFB sobre el crecimiento, proliferación y supervivencia de las células epiteliales dentales. Células epiteliales dentales fueron cultivadas en DMEM/F12, en presencia y ausencia de SFB. Observaciones morfológicas e inmunohistoquímicos de la actina, vimentina y fibronectina fueron realizados. Los resultados permitieron constatar que la ausencia de SFB afectó negativamente la proliferación de las células epiteliales dentales, pero mantuvo una expresión optima de la actina y vimentina. Se identificó una alteración en la expresión de la fibronectina en las células tratadas en ausencia de SFB. En conclusión, la carencia de SFB disminuyo drásticamente la supervivencia, proliferación y expresión de la fibronectina en las células epiteliales dentales de rata.

Reduced body weight at weaning followed by increased post-weaning growth rate interacts with part-per-trillion fetal serum concentrations of bisphenol A (BPA) to impair glucose tolerance in male mice.

There is evidence from longitudinal studies that being light at birth and weaning is associated with subsequent rapid weight gain in infants. This is referred to as “centile crossing”, which can lead to increased risk of lifetime obesity, glucose dysregulation and type 2 diabetes. Here, pregnant CD-1 mice were hemi-ovariectomized so that the entire litter was contained in one uterine horn to increase variability in fetal growth rate. Pregnant females were implanted on gestation day (GD) 9 with a Silastic capsule containing 6, 60 or 600 μg bisphenol A (BPA). On GD 18 the mean fetal serum unconjugated BPA concentrations were 17, 177 and 1858 pg/ml, respectively. Capsules were not removed, to avoid maternal stress, and were predicted to release BPA for at least 3 weeks. Body weight at weaning was strongly negatively correlated with post-weaning weight gain in both control and BPA-treated male mice, consistent with human data; female offspring were excluded, avoiding complications associated with postpubertal estrogens. Within each treatment group, male offspring were sorted into tertiles based on relative weight gain during the two weeks after weaning, designated as having Rapid (R), Medium (M) or Slow (S) growth rate. BPA exposure was associated with altered growth rate between weaning and postnatal week 12 (young adulthood), when a low-dose (20 mg/kg, i.p.) glucose tolerance test (GTT) was performed. We found altered glucose regulation in response to all doses of BPA. However, glucose tolerance was only significantly impaired (blood glucose levels were elevated) compared to controls in males in the rapid post-weaning growth group exposed perinatally to BPA. We conclude that male mice that are light at weaning, but then experience rapid catch-up growth immediately after weaning, represent a sensitive sub-population that is vulnerable to the metabolic disrupting effects of very low pg/ml fetal serum concentrations of BPA.

PDGFRα monoclonal antibody: Assessment of embryo-fetal toxicity and time-dependent placental transfer of a murine surrogate antibody of olaratumab in mice.

Olaratumab (Lartruvo™) is a recombinant human IgG1 monoclonal antibody that specifically binds PDGFRα. The maternal and in utero embryo-fetal toxicity and toxicokinetics of a human anti-mouse PDGFRα antibody (LSN3338786) were investigated in pregnant mice. A pilot study was used to set doses for the definitive study. In the definitive study, mice were administered vehicle, 5, 50, or 150 mg/kg LSN3338786 by intravenous injection on gestation days (GD) 6, 9, 12, and 15. Fetal tissues and/or serum samples were collected on GD 10, 12, 15, and 18 to evaluate exposure of antibody. There were no adverse maternal effects at 50 and 150 mg/kg although maternal deaths and adverse clinical signs were observed at 5 mg/kg. LSN3338786 crossed the placenta as early as GD 10 during organogenesis. Elimination half-life of LSN3338786 in dams decreased between GD 6 and 15. On GD 18, fetal serum concentrations of antibody were substantially higher than maternal serum concentrations at all doses. Increased incidences of malformations consisting of open and partially open eye and increased incidences of skeletal variation frontal/parietal additional ossification site occurred in fetuses from mid- and high-dose groups. The majority of transplacental migration of antibody occurred in concert with rapid maternal serum clearance before parturition. The no-observed effect level for teratogenicity of 5 mg/kg was associated with GD 15 maternal serum concentrations 3-11 times lower than clinical exposure of olaratumab, suggesting that olaratumab may cause fetal harm when administered to pregnant women.

A new epithelial cell line, SSK from kidney of striped snakehead Channa striatus (Bloch 1793)

A cell line was established from striped snakehead Channa striatus kidney. The cell line, named as SSK, has been passaged for over 62 times. Growth studies at different temperatures and foetal bovine serum (FBS) concentrations revealed that SSK cells show optimum growth at 28°C in L-15 medium containing 20% FBS. The chromosome number of SSK cells was 2n=40. Partial sequencing of mitochondrial cytochrome oxidase 1 gene of the cells confirmed that the cell line originated from C. striatus. The SSK cells were primarily epithelial, as determined using immunophenotyping. The cells from SSK cell line were transfected with phrGFP II-N mammalian expression vector. The SSK cell line was stored in liquid nitrogen (-196οC) at different passages and was successfully revived after four months of storage. The cell line could be successfullyemployed for cytotoxicity studies, as revealed by neutral red assay. The cell line can be a valuable surrogate to the whole fish studies.

Different roles of Akt and mechanistic target of rapamycin in serum‑dependent chondroprotection of human osteoarthritic chondrocytes.

Despite various animal serums being used widely to culture chondrocytes, the regulatory mechanism of serum on chondrocyte activities has not been elucidated. In the present study, human osteoarthritis (OA) chondrocytes were used to perform invitro investigations on the effect of different concentrations of bovine fetal serum on extracellular matrix synthesis, cell proliferation and autophagy using the Cell Counting Kit‑8 analysis, a laser‑scanning confocal microscope, and western blot analysis. The results demonstrated that 5% serum exerted a chondroprotective effect more than the other concentrations of serum, as it simultaneously promoted cell proliferation, autophagy, and ECM synthesis in human OA chondrocytes. Furthermore, the decreased mechanistic target of rapamycin (mTOR) and increased Akt were observed in 5% serum‑treated OA chondrocytes. Either mTOR or Akt inhibitor influenced the effect of 5% serum on cell proliferation and autophagy in human OA chondrocytes, which was associated with LC‑3B or B‑cell lymphoma-2 (Bcl‑2) signal molecules. Consistent with previous studies, the present study proposes that 5% serum promotes cell proliferation via the Akt/Bcl‑2 axis and induces autophagy via the mTOR/LC‑3B axis in human OA chondrocytes. Furthermore, the different roles of Akt and mTOR in the cell processes of human OA chondrocytes require consideration for preclinical and clinical therapy of OA.

Gestational 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126) exposure disrupts fetoplacental unit: Fetal thyroid-cytokines dysfunction

Exposure to polychlorinated biphenyls (PCBs) is related to several endocrine disorders. This study examined the effect of maternal exposure of 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126) on the fetoplacental unit and fetal thyroid-cytokine axis during the pregnancy. Pregnant albino rats received PCB 126 (20 or 40μg/kgb.wt.) by oral gavage from gestation day (GD) 1 to 20. Potential effects of PCB 126 were evaluated by following the histopathological changes in the placenta by Haematoxylin and Eosin (H&E) stain and measuring the maternofetal thyroid axis (ELIZA), maternofetal body weight, and fetal growth markers (ELIZA), and cytokines (ELIZA) at embryonic day (ED) 20. Placental tissues of both treated groups showed hyperemia, hemorrhage, degeneration and apoptosis in labyrinth layer and spiral artery at GD 20. Both administrations of PCB 126 elevated serum thyrotropin (TSH) concentration, and decreased free thyroxine (FT4) and free triiodothyronine (FT3) concentrations, resulting in a maternofetal hypothyroidism. The presence of hypothyroidism increased fetal serum concentration of transforming growth factor-β (TGF-β), leptin (LEP), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and decreased the fetal serum insulin growth factor-I (IGF-I), IGF-II, insulin, adiponectin (ADP), and growth hormone (GH) in both treated groups at ED 20. However, the increase in resistin (RETN) and interferon-γ (IFN-γ) was non-significant in low-dose group and highly significant in high-dose group. Simultaneously, the reduction in body weight of the dams and fetuses was observed in both PCB 126 groups of examined day with respect to the control group. The maternal PCB 126 distorted the fetoplacental unit might disrupt the fetal thyroid-cytokines axis and prenatal development.

Fetal DNA hypermethylation in tight junction pathway is associated with neural tube defects: A genome-wide DNA methylation analysis

ABSTRACTNeural tube defects (NTDs) are a spectrum of severe congenital malformations of fusion failure of the neural tube during early embryogenesis. Evidence on aberrant DNA methylation in NTD development remains scarce, especially when exposure to environmental pollutant is taken into consideration. DNA methylation profiling was quantified using the Infinium HumanMethylation450 array in neural tissues from 10 NTD cases and 8 non-malformed controls (stage 1). Subsequent validation was performed using a Sequenom MassARRAY system in neural tissues from 20 NTD cases and 20 non-malformed controls (stage 2). Correlation analysis of differentially methylated CpG sites in fetal neural tissues and polycyclic aromatic hydrocarbons concentrations in fetal neural tissues and maternal serum was conducted. Differentially methylated CpG sites of neural tissues were further validated in fetal mice with NTDs induced by benzo(a)pyrene given to pregnant mice. Differentially hypermethylated CpG sites in neural tissues from 17 genes and 6 pathways were identified in stage 1. Subsequently, differentially hypermethylated CpG sites in neural tissues from 6 genes (BDKRB2, CTNNA1, CYFIP2, MMP7, MYH2, and TIAM2) were confirmed in stage 2. Correlation analysis showed that methylated CpG sites in CTNNA1 and MYH2 from NTD cases were positively correlated to polycyclic aromatic hydrocarbon level in fetal neural tissues and maternal serum. The correlation was confirmed in NTD-affected fetal mice that were exposed to benzo(a)pyrene in utero. In conclusion, hypermethylation of the CTNNA1 and MYH2 genes in tight junction pathway is associated with the risk for NTDs, and the DNA methylation aberration may be caused by exposure to benzo(a)pyrene.

Bone Tissue Engineering Under Xenogeneic-Free Conditions in a Large Animal Model as a Basis for Early Clinical Applicability.

For decades, researchers have been developing a range of promising strategies in bone tissue engineering with the aim of producing a significant clinical benefit over existing therapies. However, a major problem concerns the traditional use of xenogeneic substances for the expansion of cells, which complicates direct clinical transfer. The study's aim was to establish a totally autologous sheep model as a basis for further preclinical studies and future clinical application. Ovine mesenchymal stromal cells (MSC) were cultivated in different concentrations (0%, 2%, 5%, 10%, and 25%) of either autologous serum (AS) or fetal calf serum (FCS). With an increase of serum concentration, enhanced metabolic activity and proliferation could be observed. There were minor differences between MSC cultivated in AS or FCS, comparing gene and protein expression of osteogenic and stem cell markers, morphology, and osteogenic differentiation. MSC implanted subcutaneously in the sheep model, together with a nanostructured bone substitute, either in stable block or moldable putty form, induced similar vascularization and remodeling of the bone substitute irrespective of cultivation of MSC in AS or FCS and osteogenic differentiation. The bone substitute in block form together with MSC proved particularly advantageous in the induction of ectopic bone formation compared to the cell-free control and putty form. It could be demonstrated that AS is suitable for replacement of FCS for cultivation of ovine MSC for bone tissue engineering purposes. Substantial progress has been made in the development of a strictly xenogeneic-free preclinical animal model to bring future clinical application of bone tissue engineering strategies within reach.

Enhanced in utero allogeneic engraftment in mice after mobilizing fetal HSCs by α4β1/7 inhibition

AbstractIn utero hematopoietic cell transplantation (IUHCT) is a novel nonmyeloablative approach that results in donor-specific tolerance and mixed allogeneic chimerism. Clinical application is limited by low levels of donor cell engraftment. Competition from endogenous hematopoietic stem cells (HSCs) for limited “space” in fetal hematopoietic organs remains a significant barrier to successful IUHCT. AMD3100, a CXCR4 inhibitor, and firategrast, an α4β1 and α4β7 integrin inhibitor (α4β1/7), have been shown to disrupt HSC retention in the postnatal hematopoietic niche. We hypothesized that maternal administration of AMD3100 and/or firategrast prior to IUHCT would mobilize endogenous HSCs from the fetal liver (FL) and result in preferential FL homing of donor HSCs and enhanced long-term engraftment following IUHCT in an allogeneic mouse model. We demonstrate that (1) both agents cross the placenta with rapidly detectable fetal serum concentrations following maternal administration; (2) firategrast treatment alone or with AMD3100 mobilizes endogenous HSCs from the FL and results in increased FL homing of donor HSCs following IUHCT; and (3) enhanced donor HSC homing following firategrast treatment translates into increased long-term multilineage donor cell engraftment. This approach highlights the potential of mobilization strategies to overcome barriers to successful engraftment and increase the clinical promise of IUHCT.

Development of the independent function of fetal thyroid glands in the dog in connection with iodothyronine concentrations in pregnant bitches, fetal fluids, and fetal serum

Thyroxine (T4) and triiodothyronine (T3) concentrations in pregnant and nonpregnant bitches were measured. The allantoic and amniotic fluid samples were collected separately in the third week of pregnancy, and fetal blood samples were collected in the fourth week of pregnancy. There was no difference between T4 results in the pregnant and nonpregnant animals, but the measured serum concentrations exceeded the healthy range for normal adults. Serum T4 concentrations were lower in the fetus than those in adults (P < 0.01). Fetal T4 concentrations continuously increased and reached 13.38 ± 6.19 nmol/L before birth. The fetal serum T4 concentrations were lower than the T4 concentrations in allantoic and amniotic fluid until the seventh week, and the fetal serum T3 concentrations were lower than those in fetal fluids throughout the pregnancy (P < 0.01). Maximum T3 concentrations in allantoic and amniotic fluid exceeded the concentrations in the fetal and maternal serum. It is conceivable that the considerable differences between maternal and fetal serum T4 concentrations in healthy animals are explained by the T4 impermeability of the placenta. Extremely high maternal T4 (193.5 nmol/L) in 1 bitch was associated with T4 concentrations under the detection limit in the fetal fluids and serum suggesting an inhibitory effect. The T4 concentrations in all the fetal fluids and serum were under the detectable concentration that can be defined by 3.0 nmol/L in that bitch. We have demonstrated that fetal thyroid glands start functioning independently at the same time as thyroid cell formation in the dog, but the overproduction of maternal T4 may have a suppressive effect on fetal iodothyronine production.

Placental Transfer of Perfluoroalkyl Substances and Associations with Thyroid Hormones: Beijing Prenatal Exposure Study.

Perfluoroalkyl substances (PFASs) have been detected in wildlife and human samples worldwide. Toxicology research showed that PFASs could interfere with thyroid hormone homeostasis. In this study, eight PFASs, fifteen PFAS precursors and five thyroid hormones were analyzed in 157 paired maternal and cord serum samples collected in Beijing around delivery. Seven PFASs and two precursors were detected in both maternal and cord sera with significant maternal-fetal correlations (r = 0.336 to 0.806, all P < 0.001). The median ratios of major PFASs concentrations in fetal versus maternal serum were from 0.25:1 (perfluorodecanoic acid, PFDA) to 0.65:1 (perfluorooctanoic acid, PFOA). Spearman partial correlation test showed that maternal thyroid stimulating hormone (TSH) was negatively correlated with most maternal PFASs (r = −0.261 to −0.170, all P < 0.05). Maternal triiodothyronin (T3) and free T3 (FT3) showed negative correlations with most fetal PFASs (r = −0.229 to −0.165 for T3; r = −0.293 to −0.169 for FT3, all P < 0.05). Our results suggest prenatal exposure of fetus to PFASs and potential associations between PFASs and thyroid hormone homeostasis in humans.

Mobilization of Host Hematopoietic Cells Increases Engraftment after in Utero Hematopoietic Cell Transplantation (IUHCT)

Background:IUHCT is a non-myeloablative non-immunosuppressive transplant approach that allows for donor cell engraftment across allogeneic barriers. It has the potential to treat a large number of congenital hematologic, genetic, and immunologic disorders. A significant barrier to the clinical application of IUHCT is the low level of donor cell engraftment secondary to competition for limited space in the developing hematopoietic niche. AMD3100, an inhibitor of the chemokine receptor CXCR4, and Firategrast, a small molecule inhibitor of the α4β1 integrin, have been shown to disrupt the retention of hematopoietic stem cells (HSCs) in postnatal bone marrow (BM) transplant models. We hypothesized that administration of these agents to a pregnant dam prior to IUHCT would mobilize endogenous fetal HSCs from the fetal liver (FL) hematopoietic niche resulting in preferential homing of donor HSCs and enhanced long-term donor cell engraftment.Methods:AMD3100 (5 mg/kg) and Firategrast (100 mg/kg) were maternally administered alone or in combination to time-dated Balb/c dams at either embryonic day 17 (E17) (pharmacokinetic studies) or E14 (endogenous HSC mobilization, donor cell homing, and long-term engraftment studies). Maternal and fetal sera were analyzed by mass spectrometry at 15, 45, 90, and 180 minutes following AMD3100 or Firategrast administration to determine serum drug concentrations. Based on these pharmacokinetic studies, FLs were harvested 60 minutes following AMD3100 and/or Firategrast administration and assessed by flow cytometry for lineage- sca-1+ c-kit+ (LSK) cells to determine endogenous HSC mobilization. Short-term homing and long-term engraftment were subsequently assessed in Balb/c fetal mice (H2Kd) intravenously injected with 10x106 B6 GFP (H2Kb) BM cells following maternal administration of AMD3100, Firategrast, or AMD3100+Firategrast (Combination). For homing analysis, the proportion of donor LSK within the donor lymphocyte compartment of the FL was measured 24 hours after IUHCT. Long-term engraftment of donor cells was assessed monthly by flow cytometry of peripheral blood (PB) through 6 months of age. Multilineage PB engraftment was assessed at 9 months of age. Statistical analyses were performed using one-way or two-way ANOVA with Bonferroni post-hoc tests.Results:Maternal administration of either agent resulted in transplancental passage with peak fetal serum concentrations occurring between 45 and 90 minutes for AMD3100, and between 15 and 90 minutes for Firategrast. Treatment with Firategrast or Combination (AMD3100+Firategrast) resulted in significant mobilization of endogenous HSCs from the FL, with Firategrast eliciting the greatest mobilization vs. control (no treatment) (2.8x103 vs. 9.6x103 HSCs/FL, p<0.05) (Figure 1). Maternal administration of Firategrast or Combination prior to IUHCT translated into 1) significantly increased short-term homing of allogeneic donor LSK in the fetal liver 24 hours after IUHCT (Control: 1.5% ± 0.10 donor LSK in donor lymphocyte population; Firategrast: 2.4% ± 0.3, p = 0.0135; Combination: 2.4% ± 0.1, p = 0.0136), and 2) sustained enhancement of long-term allogeneic engraftment through 6 months of age (Figure 2). There was no apparent benefit to Combination treatment over Firategrast treatment alone. All chimeric mice demonstrated multilineage engraftment at 9 months of age supporting engraftment at the level of the HSC or early progenitor cell following IUHCT.Conclusion:Maternal administration of antagonists of the CXCR4/SDF-1a and a4b1/VCAM-1 pathways results in mobilization of endogenous fetal HSCs from the FL hematopoietic niche and preferential homing of donor HSCs to the FL when administered prior to IUHCT. The combination of agents targeting both the CXCR4/SDF-1a and a4b1/VCAM-1 pathways in the current study did not yield a synergistic effect on mobilization and engraftment. This work demonstrates the potential of mobilizing endogenous fetal HSCs as a strategy to overcome barriers to successful engraftment in utero bringing IUHCT closer to clinical application. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Critical Parameters of the In Vitro Method of Vascular Smooth Muscle Cell Calcification.

BackgroundVascular calcification (VC) is primarily studied using cultures of vascular smooth muscle cells. However, the use of very different protocols and extreme conditions can provide findings unrelated to VC. In this work we aimed to determine the critical experimental parameters that affect calcification in vitro and to determine the relevance to calcification in vivo.Experimental Procedures and ResultsRat VSMC calcification in vitro was studied using different concentrations of fetal calf serum, calcium, and phosphate, in different types of culture media, and using various volumes and rates of change. The bicarbonate content of the media critically affected pH and resulted in supersaturation, depending on the concentration of Ca2+ and Pi. Such supersaturation is a consequence of the high dependence of bicarbonate buffers on CO2 vapor pressure and bicarbonate concentration at pHs above 7.40. Such buffer systems cause considerable pH variations as a result of minor experimental changes. The variations are more critical for DMEM and are negligible when the bicarbonate concentration is reduced to ¼. Particle nucleation and growth were observed by dynamic light scattering and electron microscopy. Using 2mM Pi, particles of ~200nm were observed at 24 hours in MEM and at 1 hour in DMEM. These nuclei grew over time, were deposited in the cells, and caused osteogene expression or cell death, depending on the precipitation rate. TEM observations showed that the initial precipitate was amorphous calcium phosphate (ACP), which converts into hydroxyapatite over time. In blood, the scenario is different, because supersaturation is avoided by a tightly controlled pH of 7.4, which prevents the formation of PO4 3--containing ACP.ConclusionsThe precipitation of ACP in vitro is unrelated to VC in vivo. The model needs to be refined through controlled pH and the use of additional procalcifying agents other than Pi in order to reproduce calcium phosphate deposition in vivo.

A simple and fast approach to confirm the presence of an intrauterine pregnancy

Oftentimes sonographic findings in early pregnancy are inconclusive, and differentiating between intrauterine and extrauterine pregnancies (IUP and EUP) is challenging. In the first trimester, alpha-fetoprotein (AFP) concentration in fetal serum is at least 1000 fold higher than in maternal serum. The goal of this study was to determine whether measurement of AFP concentration in material evacuated from the uterus can assist with confirmation of an IUP.

Fabrication of biocompatible lab‐on‐chip devices for biomedical applications by means of a 3D‐printing process

A new microfluidic assembly method for semiconductor‐based biosensors using 3D‐printing technologies was proposed for a rapid and cost‐efficient design of new sensor systems. The microfluidic unit is designed and printed by a 3D‐printer in just a few hours and assembled on a light‐addressable potentiometric sensor (LAPS) chip using a photo resin. The cell growth curves obtained from culturing cells within microfluidics‐based LAPS systems were compared with cell growth curves in cell culture flasks to examine biocompatibility of the 3D‐printed chips. Furthermore, an optimal cell culturing within microfluidics‐based LAPS chips was achieved by adjusting the fetal calf serum concentrations of the cell culture medium, an important factor for the cell proliferation.

The effects of maternal vitamin D on neonatal growth parameters.

Vitamin D facilitates calcium absorption and bone building. Presence of vitamin D is highly important in pregnant women due to its effect on the development of the fetal skeleton. The study population comprised 208 low-risk pregnant women of a heterogeneous population. Maternal and fetal serum concentrations of vitamin D were measured using the Liaison 25(OH)D Assay (DiaSorin, Italy). Maternal vitamin D serum concentrations correlate with neonatal vitamin D serum concentrations but do not affect neonatal weight and/or head circumference. WHAT IS KNOWN? • Vitamin D is known to be also involved in immunomodulation and cellular proliferation and differentiation. • Vitamin D is highly important in pregnant women for its effect on fetal musculoskeletal and neurological development. WHAT IS NEW? • No association was detected between maternal or neonatal vitamin D concentration with neonatal growth parameters or obstetrical complications, and no association was found between maternal vitamin D serum concentrations and maternal obstetrical complication rate. • A strong correlation was demonstrated between maternal and neonatal serum vitamin D concentrations.

Theophylline Pharmacokinetics in Foetal Sheep: Maternal Metabolic Capacity is the Principal Driver.

Understanding theophylline pharmacokinetics (PK) in the foetus is essential to prevent in utero toxicity and optimize prophylactic therapies. Previous studies in pregnancy have been obfuscated by maternal dosing and inadequate sampling in the foetus; both render modelling of foetal PK difficult. Six ewes carrying singleton foetuses received theophylline (60mg) into the foetal jugular vein. Blood samples were drawn from the foetus and ewe over 36hr. Serum concentrations were measured. Maternal and foetal pharmacokinetic parameters were estimated. Foetal non-compartmental pharmacokinetic parameters were as follows: half-life 7.37±1.22hr; volume of distribution 44.62±11.45L; area under the curve 14.82±2.71hr/(μg/mL); and clearance 4.15±0.70L/hr. Rapid theophylline distribution across the placenta was observed. Maternal non-compartmental pharmacokinetic parameters were as follows: half-life 6.54±2.44hr; volume of distribution 32.48±9.99L; area under the curve 16.28±4.53hr/(μg/mL); and clearance 3.69±1.47L/hr. Foetal and ewe serum concentration-time profiles were fit together into a 3-compartment population pharmacokinetic model, and parameters were as follows: central volume 1.38±0.11L; 2nd peripheral compartment volume 3.11±0.29L; 3rd peripheral compartment volume 60.14±6.02L; elimination clearance 9.89±0.90L/hr; distribution clearance between central and 2nd compartment 30.87±2.31L/hr; and distribution clearance between 2nd and 3rd compartments 13.89±1.11L/hr. Cytochrome P4501A expression was robust in maternal liver; negligible activities were observed in placenta, foetal liver and foetal kidney. In vitro protein binding of theophylline was 30% lower in foetal serum compared to maternal serum (29.7±4.4 versus 42.0±3.6%-bound). Free concentrations were lower in the foetus than in the ewe, suggesting active transport across placenta. In summary, foetal clearance of theophylline is attributable to rapid distribution into the maternal circulation across the placenta followed by greater maternal protein binding and metabolic activity.

Caffeine intake during pregnancy and adverse birth outcomes: a systematic review and dose-response meta-analysis.

Caffeine is commonly consumed during pregnancy, crosses the placenta, with fetal serum concentrations similar to the mother's, but studies of birth outcome show conflicting findings. We systematically searched Medline and Embase for relevant publications. We conducted meta-analysis of dose-response curves for associations between caffeine intake and spontaneous abortion, stillbirth, preterm delivery, low birth weight and small for gestational age (SGA) infants. Meta-analyses included 60 unique publications from 53 cohort and case-control studies. An increment of 100g caffeine was associated with a 14% (95% CI 10-19%) increase in risk of spontaneous abortion, 19% (5-35%) stillbirth, 2% (-2 to 6%) preterm delivery, 7% (1-12%) low birth weight, and 10% (95% CI 6-14%) SGA. There was substantial heterogeneity in all models, partly explained by adjustment for smoking and previous obstetric history, but not by prospective assessment of caffeine intake. There was evidence of small-study effects such as publication bias. Greater caffeine intake is associated with an increase in spontaneous abortion, stillbirth, low birth weight, and SGA, but not preterm delivery. There is no identifiable threshold below which the associations are not apparent, but the size of the associations are generally modest within the range of usual intake and are potentially explained by bias in study design or publication. There is therefore insufficient evidence to support further reductions in the maximum recommended intake of caffeine, but maintenance of current recommendations is a wise precaution.