Fetal-fluid proteome analyses in late-term healthy pregnant mares and in mares with experimentally induced ascending placentitis

2019 ◽  
Vol 31 (9) ◽  
pp. 1486 ◽  
Author(s):  
Igor F. Canisso ◽  
Shavahn Loux ◽  
Kirsten E. Scoggin ◽  
Edward L. Squires ◽  
Mats H. Troedsson ◽  
...  

Characterisation of fetal fluids in healthy and disease states of pregnant mares can help to unravel the pathophysiology and to identify putative markers of disease. Thus, this study aimed to compare the protein composition of: (1) amniotic and allantoic fluids of healthy mares obtained immediately after euthanasia and (2) allantoic fluid harvested via centesis before and after experimental induction of placentitis via transcervical inoculation of Streptococcus equi ssp zooepidemicus in healthy mares. Fetal fluids were analysed with a high-throughput proteomic technique after in-gel digestion. Statistical comparisons were performed following normalisation of peptide spectral match. Global normalisation was performed to calculate relative expression. There were 112 unique proteins present in both allantoic and amniotic fluids. There were 13 and 29 proteins defined as amniotic- or allantoic-specific respectively that were present in at least two fluid samples. Another 26 proteins were present in both amniotic and allantoic fluids. Panther DB functional classification grouped fetal-fluid proteins as transfer carriers, signalling molecules, receptors, immunity, hydrolase, enzymes, membrane traffic, cytoskeleton, cell adhesion, calcium binding and extracellular matrix. Experimentally induced placentitis resulted in 10 proteins being upregulated and 10 downregulated in allantoic fluid. Newly identified proteins and changes in the fetal-fluid proteome provide clues about the physiology of pregnancy and pathogenesis of placentitis.

2005 ◽  
Vol 17 (2) ◽  
pp. 177 ◽  
Author(s):  
C. Morrow ◽  
M. Berg ◽  
R. McDonald ◽  
D. Wells ◽  
A. Peterson ◽  
...  

Abnormal placentation, pregnancy failure, and hydroallantois are associated with somatic cell nuclear transfer (SCNT) in cattle. Identification of diagnostic markers for abnormal placentation in early gestation would permit therapeutic intervention. Ultrasonography and transvaginal sampling of amniotic and/or allantoic (fetal) fluid enables regular monitoring of fetal health. We report on the composition of serial samples of fetal fluid from individual cows between Days 70–130 of gestation and the potential of steroid and electrolyte composition as an early diagnostic marker for the subsequent occurrence of hydroallantois in SCNT pregnancies in cattle. On Day 70, pregnancy rates were 50% and 60% for cows or heifers implanted with single in vitro-fertilized (IVP, 20/40) or SCNT (25/42) embryos, respectively, and 67% for pregnancies generated by artificial insemination (AI, 12/18). Resulting fetuses were either clones (SCNT) or offspring (IVP/AI) of a donor Holstein bull. Fetal fluids, sampled using ultrasound-guided transvaginal puncture, were collected on Days 70, 100, and 130 of gestation (n = 12 and 139 for amniotic and allantoic samples, respectively). Placental and fetal morphological data were collected following slaughter between Days 135–163 of gestation (n = 14, 20, and 10 for SCNT, IVP, and AI groups, respectively). Fetal fluids were analyzed for progesterone, estrone sulphate, sodium, chloride, potassium, creatinine, urea, calcium, magnesium and phosphate. Pregnancy outcomes for the SCNT group were retrospectively classified as: Fail 100 (pregnancies failing between Days 70–99; n = 6); Fail 130 (failing between Days 100–129; n = 5); Hydrops (greater than 10 L combined amniotic and allantoic fluid at postmortem between Days 135–163; n = 8) and SCNT Pregnant 150 (pregnant between Days 135–163; n = 6). IVP and AI pregnancies were classified as IVP or AI Pregnant 150. Fluid composition was analyzed by ANOVA on log-transformed data. On Day 70, allantoic progesterone and estrone sulphate concentrations were significantly higher (P < 0.05) for the SCNT cows compared to the IVP/AI Pregnant 150 cows. On Day 70, allantoic potassium, chloride, creatinine, and urea concentrations were significantly higher (P < 0.05) for the SCNT Hydrops cows compared to the IVP/AI Pregnant 150 cows. In addition, Day 70 allantoic creatinine and urea concentrations were significantly higher (P < 0.05) for the SCNT Hydrops cows compared to other SCNT groups. By Day 100, allantoic chloride, creatinine, and urea concentrations in SCNT Hydrops cows were significantly lower (P < 0.05) than in IVP/AI Pregnant 150 groups. We conclude that elevated Day 70 allantoic urea and creatinine concentrations are potential early diagnostic markers predicting hydroallantois in recipient cattle carrying SCNT fetuses. Further investigation of these markers in other somatic donor cell lines used for nuclear transfer is warranted to determine their general utility.


2018 ◽  
Vol 206 (4-5) ◽  
pp. 208-217
Author(s):  
Debora Kehl ◽  
Sabrina Görtz ◽  
Craig Wang ◽  
Simon P. Hoerstrup ◽  
Ulrich Bleul ◽  
...  

The ovine developmental model represents the standard in vivo model for studies involving maternofetal physiology, amniotic fluid (AF) research, and fetal cell therapy prior to human clinical use. Although being close to the human fetal anatomy, 2 separate extraembryonic fluid compartments remain during gestation, known as the amnion and the allantois. A clear distinction between AF versus allantoic fluid (AL) is therefore indispensable for correct scientific conclusions with regard to human translation. In the presented study, the biochemical composition of AF and AL was evaluated in ovine gravid uteri postmortem (n = 31) over the entire gestation. Four parameters, consisting of Na+, Cl–, Mg2+, and total protein, have been found to allow for specific discrimination of the 2 fetal fluids at all gestational phases and therefore as potential surrogate parameters for gestational age. In addition, volumetric changes of the developing fetus and the 2 fetal fluid cavities were analyzed by contrast-enhanced computed tomography (n = 12). AF showed a significant, linear volumetric increase over gestation, whereas AL volume maintained relatively static independent of gestational age. These results serve as a basis for future studies by providing surrogate markers enabling a reliable distinction of isolated fetal fluids and contained cells in the ovine developmental model over the entire gestation.


1982 ◽  
Vol 208 (1) ◽  
pp. 239-242 ◽  
Author(s):  
D C Wraith ◽  
C J Chesterton

Current evidence suggests that the major cell-surface modification occurring during mammalian erythropoiesis could be generated by two separate mechanisms: either selective loss of membrane proteins during enucleation or endocytosis at the subsequent reticulocyte and erythrocyte stages. The former idea was tested by collecting developing rabbit erythroid cells before and after the enucleation step and comparing their cell-surface protein composition via radiolabelling and electrophoresis. Few changes were observed. Our data thus lend support to the endocytosis mechanism.


PeerJ ◽  
2019 ◽  
Vol 6 ◽  
pp. e6228 ◽  
Author(s):  
Andrés Montoya ◽  
Manuel Carlos López ◽  
Ivan D. Vélez ◽  
Sara M. Robledo

Background Leishmaniasis is a parasitic disease caused by more than 20 species of the Leishmania genus. The disease is globally distributed and is endemic in 97 countries and three territories in the tropical and subtropical regions. The efficacy of the current treatments is becoming increasingly low either due to incomplete treatment or resistant parasites. Failure of treatment is frequent, and therefore, the search for early biomarkers of therapeutic response in cutaneous leishmaniasis (CL) is urgently needed. Objective The aim of this study was to compare the proteomic profiles in patients with CL before and after 7 days of treatment and identify early biomarkers of curative response. Methods Four patients with a parasitological diagnosis of leishmaniasis with confirmation of species by PCR-RFLP were recruited. All patients had a single lesion, and a protein from the middle of the ulcer was quantified by liquid chromatography and mass spectrometry. Results A total of 12 proteins showed differential expression in the comparative LC-electrospray ionization MS/MS (LC-ESI-MS/MS) triplicate analysis. Seven of them were up-regulated and five of them were down-regulated. Calcium binding proteins A2, A8, and A9 and hemoglobin subunits alpha-2 and delta showed high correlation with epidermis development and immune response. Conclusion We identified changes in the profiles of proteins that had a positive therapeutic response to the treatment. The proteins identified with differential expression are related to the reduction of inflammation and increased tissue repair. These proteins can be useful as biomarkers for early monitoring of therapeutic response in CL.


Reproduction ◽  
2004 ◽  
Vol 127 (1) ◽  
pp. 57-66 ◽  
Author(s):  
J A McGlothlin ◽  
G D Lester ◽  
P J Hansen ◽  
M Thomas ◽  
L Pablo ◽  
...  

An experimental model of ascending placentitis was developed in the mare to characterize the uterine myoelectrical pattern in late gestation and determine how ascending placentitis altered this pattern. In experiment 1, myometrial electrical activity was analyzed during the early morning, late morning and evening hours in four mares in the last 15 days of gestation to identify patterns of activity. In experiment 2, nine mares received intra-cervical inoculations ofStreptococcus equisubspecieszooepidemicus. Myoelectrical activity in the early morning and evening hours in these mares was compared with four control mares. In experiment 1, the number of spike burst clusters >30 s was greater in the evening than in the late morning hours (P< 0.04). Spike burst activity (number × duration) of mares in experiment 1 was similar during day and night recordings until the last 6 days of gestation when it gradually increased each evening until parturition (P< 0.05). In experiment 2, control mares experienced a gradual increase in the number of small spike burst clusters in the last 6 days (P= 0.008) and an increase in large and small spike burst clusters in the evening hours in the last 4 days of gestation (P= 0.03). Mares with experimentally induced placentitis never exhibited a rise in spike burst clusters but had an increase in the mean duration and activity index of large spike burst clusters in the 4 days before parturition (P< 0.04). In conclusion, control mares had a progressive, reversible rise in myoelectrical activity at night in the week preceding parturition. This was not observed in mares with experimentally induced placentitis. They exhibited an increase in the intensity and duration of large spike burst clusters possibly in response to local inflammation.


2010 ◽  
Vol 104 (5) ◽  
pp. 693-700 ◽  
Author(s):  
Robin F. J. Benus ◽  
Tjip S. van der Werf ◽  
Gjalt W. Welling ◽  
Patricia A. Judd ◽  
Moira A. Taylor ◽  
...  

The intestinal microbiota are a complex ecosystem influencing the immunoregulation of the human host, providing protection from colonising pathogens and producing SCFA as the main energy source of colonocytes. Our objective was to investigate the effect of dietary fibre exclusion and supplementation on the intestinal microbiota and SCFA concentrations. Faecal samples were obtained from healthy volunteers before and after two 14 d periods of consuming formulated diets devoid or supplemented with fibre (14 g/l). The faecal microbiota were analysed using fluorescentin situhybridisation and SCFA were measured using GLC. There were large and statistically significant reductions in the numbers of theFaecalibacterium prausnitzii(P ≤ 0·01) andRoseburiaspp. (P ≤ 0·01) groups during both the fibre-free and fibre-supplemented diets. Significant and strong positive correlations between the proportion ofF. prausnitziiand the proportion of butyrate during both baseline normal diets were found (pre-fibre freer0·881,P = 0·001; pre-fibre supplementedr0·844,P = 0·002). A significant correlation was also found between the proportional reduction inF. prausnitziiand the proportional reduction in faecal butyrate during both the fibre-free (r0·806;P = 0·005) and the fibre-supplemented diet (r0·749;P = 0·013). These findings may contribute to the understanding of the association between fibre, microbiota and fermentation in health, during enteral nutrition and in disease states such as Crohn's disease.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Paulo Cezar Rocha dos Santos ◽  
Claudine J. C. Lamoth ◽  
Fabio Augusto Barbieri ◽  
Inge Zijdewind ◽  
Lilian Teresa Bucken Gobbi ◽  
...  

Abstract We examined the effects of age on intermuscular beta-band (15–35 Hz) coherence during treadmill walking before and after experimentally induced fatigue. Older (n = 12) and younger (n = 12) adults walked on a treadmill at 1.2 m/s for 3 min before and after repetitive sit-to-stand, rSTS, to induce muscle fatigability. We measured stride outcomes and coherence from 100 steps in the dominant leg for the synergistic (biceps femoris (BF)-semitendinosus, rectus femoris (RF)-vastus lateralis (VL), gastrocnemius lateralis (GL)-Soleus (SL), tibialis anterior (TA)-peroneus longus (PL)) and for the antagonistic (RF-BF and TA-GL) muscle pairs at late swing and early stance. Older vs. younger adults had 43–62% lower GL-SL, RF-VL coherence in swing and TA-PL and RF-VL coherence in stance. After rSTS, RF-BF coherence in late swing decreased by ~ 20% and TA-PL increased by 16% independent of age (p = 0.02). Also, GL-SL coherence decreased by ~ 23% and increased by ~ 23% in younger and older, respectively. Age affects the oscillatory coupling between synergistic muscle pairs, delivered presumably via corticospinal tracts, during treadmill walking. Muscle fatigability elicits age-specific changes in the common fluctuations in muscle activity, which could be interpreted as a compensation for muscle fatigability to maintain gait performance.


Reproduction ◽  
2002 ◽  
pp. 259-265 ◽  
Author(s):  
◽  
Y Xia ◽  
T O'Shea ◽  
S Hayward ◽  
AE O'Connor ◽  
...  

The aim of this study was to investigate the changes in follistatin, an activin binding protein, during the oestrous cycle, gestation and parturition in ewes using a radioimmunoassay for total follistatin, which uses dissociating reagents to remove the interference of activin. Follistatin concentrations remained unchanged (2.7 +/- 0.2 ng ml(-1)) during the oestrous cycle and decreased as pregnancy progressed. Follistatin concentrations in allantoic fluid also decreased during gestation, whereas in amniotic fluid follistatin concentrations reached a peak at day 75 of gestation (9.8 ng ml(-1)) and had decreased to 4.4 ng ml(-1) at day 140. Follistatin concentrations in fetal blood (7.0 +/- 0.5 ng ml(-1)) did not change from day 50 to day 140 of gestation but were significantly higher than in matched maternal samples (3.1 +/- 0.3 ng ml(-1)). Circulating follistatin in ewes was significantly increased on the day of parturition (5.6 +/- 0.6 ng ml(-1)) compared with the days before parturition (2.7 +/- 0.4 ng ml(-1)), but had decreased by day 2 after birth. Blood samples from newborn lambs showed that plasma follistatin concentration (13.4 +/- 2.3 ng ml(-1)) was significantly higher than that of the mothers and remained high for at least 7 days after birth. These data support previous studies of the human menstrual cycle indicating that follistatin is not an endocrine signal from the ovary; however, in contrast to human pregnancies, follistatin concentrations in sheep decreased and become high only after or during parturition. This difference observed between species may reflect different physiological effects of follistatin or may be the result of measurement of different isoforms.


1985 ◽  
Vol 106 (3) ◽  
pp. 355-360 ◽  
Author(s):  
H. A. Robertson ◽  
R. J. Dwyer ◽  
G. J. King

ABSTRACT In order to correlate the concentrations of oestrogens in the fetal fluids of the pig with those observed in the maternal blood and urine, changes in the concentrations of oestrone, oestradiol-17β, oestrone sulphate, oestradiol sulphates and oestrone glucuronide were assessed throughout pregnancy in the fetal and maternal fluids. In general, the pattern of change was similar for all oestrogens measured in both fetal and maternal fluids. Since the concentration of oestrogens in allantoic fluid during early pregnancy is reflected in the concentration of these steroids in maternal plasma and excreted in the maternal urine, the rise and fall of oestrogen concentrations around day 30 is suggestive of synthesis followed by a virtual cessation of oestrogen production until the fetus or placenta again produce increasing amounts detectable after day 45. These findings contrast sharply with those in the cow and the ewe where, although similar peaks in oestrogen concentrations are observable in allantoic fluid during early pregnancy, they are not reflected in blood. J. Endocr. (1985) 106, 355–360


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