embryonic death
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2021 ◽  
Vol 12 ◽  
Author(s):  
Li Duan ◽  
Xiao Xu ◽  
Limei Xu ◽  
Caining Wen ◽  
Kan Ouyang ◽  
...  

Proteolytic targeting chimeras (PROTACs) is a rapid and reversible chemical knockout method. Compared with traditional gene-editing tools, it can avoid potential genetic compensation, misunderstandings caused by spontaneous mutations, or gene knockouts that lead to embryonic death. To study the role of estrogen receptor alpha (ERα) in the occurrence and progression of menopausal arthritis, we report a chemical knockout strategy in which stable peptide-based (PROTACs) against ERα to inhibit their function. This chemical knockdown strategy can effectively and quickly inhibit ERα protein in vivo and in vitro. In the rat menopausal arthritis model, this study showed that inhibiting estrogen function by degrading ERα can significantly interfere with cartilage matrix metabolism and cause menopausal arthritis by up-regulating matrix metalloproteinase (MMP-13). The results of this study indicate that ERα is a crucial estrogen receptor for maintaining cartilage metabolism. Inhibition of ERα function by PROTACs can promote the progression of osteoarthritis.


2021 ◽  
Author(s):  
Dilek Cicek ◽  
Nick Warr ◽  
Gozde Yesil ◽  
Hatice Kocak Eker ◽  
Firdevs Bas ◽  
...  

Context: Homozygous and heterozygous variants in PPP2R3C are associated with syndromic 46,XY complete gonadal dysgenesis (MEGD syndrome), and impaired spermatogenesis, respectively. This study expands the role of PPP2R3C in the aetiology of gonadal dysgenesis (GD).  Method: We sequenced the PPP2R3C gene in four new patients from three unrelated families. The clinical, laboratory and molecular characteristics were investigated. We have also determined the requirement for Ppp2r3c in mice (C57BL6/N) using CRISPR/Cas9 genome editing. Results: A homozygous c.578T>C (p.L193S) PPP2R3C variant was identified in one 46,XX girl with primary gonadal insufficiency, 2 girls with 46,XY complete GD, and one undervirilized boy with 46,XY partial GD. The patients with complete GD had low gonadal and adrenal androgens, low AMH, and high FSH and LH concentrations. All patients manifested characteristic features of MEGD syndrome. Heterozygous Ppp2r3c knockout mice appeared overtly normal and fertile. Inspection of homozygous embryos at 14.5, 9.5 and 8.5 days post coitum revealed evidence of dead embryos. We conclude that loss of function of Ppp2r3c is not compatible with viability in mice and results in embryonic death from 7.5 dpc or earlier. Conclusion: Our data indicate essential roles for PPP2R3C in mouse and human development. Germline homozygous variants in human PPP2R3C are associated with distinctive syndromic GD of varying severity in both 46,XY and 46,XX individuals.


2021 ◽  
Vol 51 (2) ◽  
pp. 205-211
Author(s):  
N. Okur ◽  
S.A. Eratalar

The aim of the research was to determine whether enrichment of the atmosphere in an incubator with carbon dioxide (CO2) and oxygen (O2) and eggshell thickness (EST) affected embryonic death (ED), hatchability of fertile eggs (HFE) and hatching time (HT). A total of 320 Ross 308 eggs were used and the experiment was repeated twice. Eggshell thickness was classified as thin (<31 μm), medium (31 - 32 μm) and thick (>32 μm). The incubators were operated with their internal atmosphere enriched with CO2 (1.57% CO2; 20.23% O2) or O2 (0.50% CO2; 22.44% O2). Embryonic death, HFE and HT data were monitored at three periods, namely early (<486 hours), middle (486 - 492 hours), and late (492 - 510 hours). Early ED, late ED and hatchability of fertile eggs were not affected by EST or by the incubator’s internal atmosphere (P >0.05). Thus, O2 supplementation to the incubator was deemed unnecessary at 822 m altitude. There was a highly significant interaction between EST and HT. Eggs with shells 31 to 32 μm thick hatched at an appreciably greater rate between 486 and 492 hours of incubation (17%) than eggs with thicker (0.6%) or thinner (0.4%) shells in both the CO2 and O2 enriched atmospheres. The hatching rate was significantly higher in the eggs with an intermediate EST than in thick-shelled eggs. A greater proportion of eggs hatched at the late HT as opposed to earlier, regardless of EST.


2021 ◽  
Vol 8 ◽  
Author(s):  
Katy A. Martin ◽  
Juli Henderson ◽  
Matthew T. Brewer

Tritrichomonas foetus is a sexually-transmitted protozoan parasite that causes early embryonic death in cattle. Tritrichomonas foetus is enzootic in the United States but is not a reportable disease at the national level. Thus, it is difficult to understand the prevalence and relative distribution of the disease for the purpose of developing appropriate control measures. In this study, a survey of state veterinarians was used to determine the number of reported cases in each state from 2015 to 2019. Our investigation revealed infections in 25 different states and a total of 3,817 reported cases nationwide. Infections occurred throughout different regions of the country, and numbers of cases were only weakly correlated with total number of cattle in each state. Tritrichomonas foetus is a significant pathogen in the United States and understanding the relative distribution of the parasite is useful for prioritizing surveillance and intervention strategies going forward.


Author(s):  
Guilherme Dutra Muniz Oliveira ◽  
Luiz Altamiro Nogueira Garcia ◽  
Luiza Aymée ◽  
Walter Lilenbaum ◽  
Guilherme Nunes de Souza

Life ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 762
Author(s):  
Yihong He ◽  
Wenxian Chen ◽  
Jindai Fan ◽  
Shuangqi Fan ◽  
Hongxing Ding ◽  
...  

Porcine parvovirus (PPV) infection is the primary cause of SMEDI (stillbirth; mummification; embryonic death; infertility) syndrome, which is a global burden for the swine industry. Thus, it is crucial to establish a rapid and efficient detection method against PPV infection. In the present work, we developed a recombinase-aided amplification (RAA) assay, coupled with a lateral flow dipstick (LFD), to achieve an amplification of PPV DNA at 37 °C within 15 min. The detection limits of PPV RAA-LFD assay were 102 copies/μL recombinant plasmid pMD19-T-VP1, 6.38 × 10−7 ng/μL PPV DNA, and 10−1 TCID50/mL virus, respectively. This method was highly specific for PPV detection with no cross-reactivity for other swine pathogens. In contrast to polymerase chain reaction (PCR), the PPV RAA-LFD assay is more sensitive and cost-saving. Hence, the established PPV RAA-LFD assay provided an alternative for PPV detection, especially in resource-limited regions.


2021 ◽  
Vol 99 (Supplement_2) ◽  
pp. 26-26
Author(s):  
Bangmin Liu ◽  
Sudikshya Paudel ◽  
Magdalina Cummings ◽  
Xiaoqiu Wang

Abstract Adrenomedullin (ADM) is an evolutionarily conserved peptide hormone that regulates implantation, embryo spacing and placentation in humans and rodents. However, the potential roles of ADM in domestic animals, particularly pigs (as litter-bearing species), are not known. This study investigated expression patterns of ADM and its receptor components that include calcitonin receptor-like receptor (CALCRL), and receptor activity modifying protein (RAMP2) in porcine uteri and conceptuses (embryonic/fetus and its extra-embryonic membranes) during the peri-implantation period of pregnancy when 30–40% of embryonic death loss occurs. Gilts were observed for estrus and/or bred via artificial insemination at 12 and 24 h after onset of estrus (Day 0). On D 10, 11, 12, 13, 14, 15, or 16 of the estrous cycle and pregnancy (n=6 gilts per day and status), uteri were flushed with 20 ml sterile PBS (pH 7.2) after gilts were subjected to a midventral laparotomy. Pregnancy was confirmed by the presence of one or more morphologically normal conceptuses. Real-time quantitative RT-PCR analyses revealed that ADM mRNA in the endometrium was elevated for pregnant than for cyclic gilts between D 10 and 16 of pregnancy (day x status, P &lt; 0.05). In cyclic gilts, endometrial ADM mRNA increased 9.0-fold (P &lt; 0.05) between D 10 and 16; whereas it increased 58.1-fold (P &lt; 0.01) in pregnant gilts between D 10 and 16. As ADM receptors, expression of mRNAs for CALCRL and RAMP2 also increased (P &lt; 0.05) 13.4- and 5.5-fold, respectively, in the porcine conceptuses between D 10 and 16 of pregnancy. Further, ELISA analyses showed that total recoverable ADM in the uterine flushings was greater (status, P &lt; 0.0001) for pregnant than for cyclic gilts in which ADM increased 173.8-fold (P &lt; 0.0001) between D 10 and 16 of pregnancy. These results indicate that ADM may play functional roles in survival, growth and development of the porcine conceptus during the peri-implantation period of pregnancy.


2021 ◽  
Vol 14 (4) ◽  
pp. 986-995
Author(s):  
Heba Roshdy ◽  
Azhar G. Shalaby ◽  
Ahmed Abd Elhalem Mohamed ◽  
Heba Badr

Background and Aim: Rabbits are a highly sensitive species and susceptible to various bacterial pathogens that may be causative agents for early embryonic death. This study aimed to explore the administration of different bacterial agents in does suffering from early embryonic death. Furthermore, identification of genes associated with virulence was performed to identify the phenotypic and genotypic antimicrobial resistance patterns that may increase the virulence of pathogens and lead to early embryonic death. Materials and Methods: We isolated and identified bacterial agents in 106 samples from live and dead female rabbits that had undergone early embryonic death, including liver and intestine tissue, aborted fetuses, discharges, and vaginal swabs. Conventional polymerase chain reaction (PCR) was conducted to confirm the identity of the isolated bacterial strains and their virulence. Moreover, antibiotic resistance was studied phenotypically and genotypically. Results: We isolated Escherichia coli, Salmonella, Staphylococcus aureus, Pasteurella multocida, and Listeria monocytogenes. PCR confirmed typical identification except in P. multocida, which was confirmed as Gallibacterium spp. in some cases. The final percentage of detection was 34%, 30.2%, 16.9%, 13.2%, and 11.3%, respectively. Virulence properties were investigated using different designated genes. All Salmonella strains harbored invA, stn, avrA, and ompf genes, while the sopE gene was identified in 31.25%. E. coli strains harboring the iss gene lacked the shiga toxin (stx1) gene. L. monocytogenes and S. aureus strains harbored the hemolysin gene (66.7% and 33.4%, respectively). Multidrug resistance was detected phenotypically and genotypically in most strains. Each bacterial pathogen had a different antibiotic resistance profile. Conclusion: Multiple bacterial species may contribute to early embryonic death in does. Furthermore, the combined infection could be the main cause of early embryonic death. Thus, monitoring programs should bear this in mind and focus on the early detection of these bacterial agents in female rabbits to avoid embryonic death.


2021 ◽  
Vol 282 (5) ◽  
pp. 720-732
Author(s):  
Francisco Acuña ◽  
Claudio G. Barbeito ◽  
Enrique L. Portiansky ◽  
María A. Miglino ◽  
Mirta A. Flamini

Author(s):  
G. Bonev

Abstract. The objectives of this study were: 1) to determine the percent of cows with low P4 at Day 7-8 post-AI; 2) to treat the animal with low progesterone for 14 days with a P4-releasing intravaginal device (PRID DELTA); 3) to analyze the influence of P4 exogenous supplementation on pregnancy rate in treated and non-treated cows and 4) to create a protocol using “P4 Rapid” test allowing increased pregnancy rate under farm conditions. For this purpose, the current study has been conducted on 155 repeat breeding Holstein cows (<3 artificially inseminated – AI) with prolonged estrus cycle (<26 days). At day 7 and 8 post-AI, the milk progesterone levels were measured (P4 Rapid test). The cows with low P4 (n=104 or 67%) were divided into two groups each n=52. In Group I the progesterone devices were inserted for two weeks and control Group II without progesterone devices. At days 40 and 50 post artificial insemination (AI) all animals (n=104) were submitted to ultrasound pregnancy detection. Pregnancy was registered in 27 animals (51.9%) for Group 1, and in 16 cows (30.7%) for Group II. P4 Rapid is a quick, easy and practically applicable test to measure milk progesterone levels in dairy farms. The method is highly accurate and detects animals with low or high P4 at Day 7-8 after insemination. It also allows identification of conditions such as ovarian cysts, ovarian hypofunction and distinguishing of cycling and non-cycling cows. The combination of P4 measurement by P4 Rapid test and treatment with exogenous P4 to increase circulating concentrations in the first few days after conception can prevent embryo death and improve reproductive efficiency in dairy cattle.


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