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Background: To identify the correlation among female age, cellular aging markers, and aneuploidy rate in in vitro fertilization (IVF) and the preimplantation genetic test for aneuploidy (PGT-A) cycles. Methods: This is a prospective cohort study recruiting 110 infertile women between August 2017 and July 2018. They were divided into young-age (<38 years, n = 60) and advanced-age (≥38 years, n = 50) groups. Peripheral leukocytes were assessed, and the granulosa cells were pooled during oocyte pickup. Mitochondrial DNA (mtDNA) copy number and telomere length (TL) were measured using real-time polymerase chain reaction. PGT-A was performed on the NGS platform. Results: mtDNA copy number and TL were positively correlated in both leukocytes (rho = 0.477, p < 0.001) and granulosa cells (rho = 0.361, p < 0.001), but the two parameters in leukocytes were not correlated with those in granulosa cells. In the young-age group, TL in the granulosa cells was the only factor correlated with the aneuploidy rate (rho = −0.283, p = 0.044), whereas in the advanced-age group, age was the main factor (rho = 0.358, p = 0.018). Conclusions: TL in the granulosa cells was negatively correlated with the aneuploidy rate in the young-age group, supporting the application of PGT-A in younger women. [ABSTRACT FROM AUTHOR]

Avoiding aneuploid embryo transfers has been shown to improve pregnancy outcomes in patients with implantation failure and pregnancy loss. This retrospective cohort study aims to analyze the correlation of time-lapse (TL)-based variables and numeric blastocyst morphological scores (TLBMSs) with different mosaic levels. In total, 918 biopsied blastocysts with time-lapse assessments at a uniform time-point were subjected to next-generation sequencing–based preimplantation genetic testing for aneuploidy. In consideration of patient- and cycle-related confounding factors, all redefined blastocyst morphology components of low-grade blastocysts, that is, expansion levels (odds ratio [OR] = 0.388, 95% confidence interval [CI] = 0.217–0.695; OR = 0.328, 95% CI = 0.181–0.596; OR = 0.343, 95% CI = 0.179–0.657), inner cell mass grades (OR = 0.563, 95% CI = 0.333–0.962; OR = 0.35, 95% CI = 0.211–0.58; OR = 0.497, 95% CI = 0.274–0.9), and trophectoderm grades (OR = 0.29, 95% CI = 0.178–0.473; OR = 0.242, 95% CI = 0.143–0.411; OR = 0.3, 95% CI = 0.162–0.554), were less correlated with mosaic levels ≤20%, <50%, and ≤80% as compared with those of top-grade blastocysts (p < 0.05). After converting blastocyst morphology grades into scores, high TLBMSs were associated with greater probabilities of mosaic levels ≤20% (OR = 1.326, 95% CI = 1.187–1.481), <50% (OR = 1.425, 95% CI = 1.262–1.608), and ≤80% (OR = 1.351, 95% CI = 1.186–1.539) (p < 0.001). The prediction abilities of TLBMSs were similar for mosaic levels ≤20% (AUC = 0.604, 95% CI = 0.565–0.642), <50% (AUC = 0.634, 95% CI = 0.598–0.671), and ≤80% (AUC = 0.617, 95% CI = 0.576–0.658). In conclusion, detailed evaluation with TL monitoring at the specific time window reveals that redefined blastocyst morphology components and converted numeric TLBMSs are significantly correlated with all of the threshold levels of mosaicism. However, the performance of TLBMSs to differentiate blastocysts with aberrant ploidy risk remains perfectible. [ABSTRACT FROM AUTHOR]

Liver cancer is one of the most common malignant tumors globally and is the second leading cause of cancer deaths in Taiwan. The activity of α‐fucosidase (AFU) in the human body has been confirmed by many clinical studies and experiments to be applicable in the early diagnosis of primary liver cancer. Herein, an electrochemical method for determining AFU activity was developed based on measuring p‐nitrophenol (pNP), which is a product of the reaction between AFU and p‐nitrophenyl‐α‐L‐fucopyranoside. The electrochemical detection of pNP in serum samples exhibited a wide linear range (2.5–250 μM), which corresponds to approximately 2.5–250 U/L AFU. The recovery of AFU measured upon enzyme addition to standard serum was 97–110%. Moreover, no potential interferants had a significant effect on pNP quantification. In contrast, an optical absorption method was subject to interference by the serum color. Further, the linear range for pNP was only 2.5–50 μM and the recovery of AFU was 165–219%. The performance of this absorption method could lead to a misjudgment of the disease course. Therefore, the electrochemical method for AFU activity determination has greater potential for use in clinical applications. [ABSTRACT FROM AUTHOR]

Chloramphenicol (CAP) is a veterinary antibiotic that has been banned due to its severe side effects in humans. Through the application of manure, veterinary antibiotics can enter the soil, where they can be taken up by crops and vegetables and pose a potential health hazard to humans. Thus, it is highly desirable to develop a rapid and sensitive tool for on‐site detection of CAP to ensure food safety and to control the abuse of antibiotics. To this end, nitrogen‐doped graphene quantum dots (N‐GQDs) were successfully prepared via microwave‐assisted synthesis using citric acid and urea as carbon and nitrogen sources, respectively. Analytical results suggested that the interaction between N‐GQDs and CAP could occurs via π‐π stacking, which quenched N‐GQD fluorescence. CAP spiked into chicken feed could be rapidly extracted with ethanol and quantified based on N‐GQD fluorescence quenching without further separation. This method showed good recovery (97–102.6%), a low detection limit (1.8 ppm), and was not affected by interference from florfenicol, and thiamphenicol, legal substitute antibiotics. This method has excellent potential for determination of CAP in livestock feed and soil. [ABSTRACT FROM AUTHOR]

Are the morphokinetics of euploid blastocysts evaluated by a generally applicable algorithm associated with the clinical outcomes of single-embryo transfer (SET)? Time-lapse microscopy was used to compare morphokinetic variables between expanded blastocysts derived from preimplantation genetic testing for aneuploidy cycles using high-resolution next-generation sequencing (hr-NGS). The clinical efficacy of the morphokinetic algorithm KIDScore D5 was evaluated after euploid SET. Compared with euploid blastocysts, low-level mosaic blastocysts presented comparable morphokinetic and morphological features. However, high-level mosaic blastocysts exhibited significant delays in t5 (median 51.9 h post insemination (hpi), P = 0.034) (where t is the time for the embryo to reach the specific stage in hours after ICSI or conventional IVF) and t8 (median 58.6 hpi, P = 0.032) accompanied by a prolonged time period for the third cell cycle (median 14.7 h, P = 0.012). A significantly higher incidence (P = 0.011) of multinucleation indicated a susceptibility of high-level mosaic blastocysts to mitotic errors. Only a delay in the time for the embryo to reach the full blastocyst stage (median 106.0 hpi, P = 0.039) was revealed in aneuploid blastocysts, reflecting the reduced formation of good-quality blastocysts (42.6% versus 65.7%, P < 0.001). Euploid blastocysts with specific morphokinetic characteristics were graded using the KIDScore D5 algorithm. Grade C embryos achieved significantly lower rates of clinical pregnancy, implantation and ongoing pregnancy (25%, 25% and 10%, respectively) compared with the grade A (76.2%, 79.4% and 68.3%, respectively) or grade B (62.5%, 66.7% and 62.5%, respectively) embryos (P = 0.0171 to <0.0001). Although morphokinetic features appear dissimilar in embryos with different diploid–aneuploid mosaic levels, predicting chromosomal abnormalities using morphokinetics alone is still insufficient. When combined with hr-NGS, use of the generally applicable KIDScore D5 algorithm has the potential to discriminate euploid blastocysts with different developmental competence. [ABSTRACT FROM AUTHOR]

The porcine epidemic diarrhoea virus (PEDV) devastates the health of piglets but may not infect piglets whose CMP-N-glycolylneuraminic acid hydroxylase (CMAH) gene is mutated (knockouts, KO) by using CRISPR/Cas9 gene editing techniques. This hypothesis was tested by using KO piglets that were challenged with PEDV. Two single-guide RNAs targeting the CMAH gene and Cas9 mRNA were microinjected into the cytoplasm of newly fertilized eggs. Four live founders generated and proven to be biallelic KO, lacking detectable N-glycolylneuraminic acid (NGNA). The founders were bred, and homozygous offspring were obtained. Two-day-old (in exps. I, n = 6, and III, n = 15) and 3-day-old (in exp. II, n = 9) KO and wild-type (WT, same ages in respective exps.) piglets were inoculated with TCID50 1x103 PEDV and then fed 20 mL of infant formula (in exps. I and II) or sow’s colostrum (in exp. III) every 4 hours. In exp. III, the colostrum was offered 6 times and was then replaced with Ringer/5% glucose solution. At 72 hours post-PEDV inoculation (hpi), the animals either deceased or euthanized were necropsied and intestines were sampled. In all 3 experiments, the piglets showed apparent outward clinical manifestations suggesting that infection occurred despite the CMAH KO. In exp. I, all 6 WT piglets and only 1 of 6 KO piglets died at 72 hpi. Histopathology and immunofluorescence staining showed that the villus epithelial cells of WT piglets were severely exfoliated, but only moderate exfoliation and enterocyte vacuolization was observed in KO piglets. In exp. II, delayed clinical symptoms appeared, yet the immunofluorescence staining/histopathologic inspection (I/H) scores of the two groups differed little. In exp. III, the animals exhibited clinical and pathological signs after inoculation similar to those in exp. II. These results suggest that porcine CMAH KO with nullified NGNA expression are not immune to PEDV but that this KO may lessen the severity of the infection and delay its occurrence. [ABSTRACT FROM AUTHOR]

Reaction of compounds [(NO)2Fe(TMEDA)] and [BH4]− affords the first hydride-containing dinitrosyl iron complex (DNIC) [(NO)2Fe(η2-BH4)]− (2). Hydride-insertion reactivity of DNIC 2 promotes the reductive transformation of CS2 into DNIC [(NO)2Fe(η3-HCS2)]−, the first DNIC featuring both linear/bent NO ligands and Fe 3dz2-to-HCS2π* backbonding interaction. [ABSTRACT FROM AUTHOR]

This research resolves an inconsistency problem that arises from assessing circularity of workpiece measured by coordinate measuring machines (CMMs). Although the notion of circularity is employed to constrain two-dimensional circular features, in practice the measured points are obtained in a three-dimensional space and are in general not in the same plane, let alone being in a 'perpendicular' cross-section. All of the algorithms currently used for assessing circularity deal with data in a single plane that is perpendicular to the axis of a cylindrical feature from which the circular feature is extracted. This discrepancy causes the assessed circularity significantly departing from the actual circularity and resulting in rejection of in-tolerance parts. This research designs a compensation procedure for deriving two-dimensional data from three-dimensional biased measured points. The circularity is assessed based on the compensated two-dimensional data points. The assessment results with the compensation are compared with those without compensation. A program containing a variety of implementations of form fitting algorithms is developed and used to illustrate the improvement on the accuracy of assessment. [DOI: 10.1115/1.1312336]

In artificial photosynthesis, water splitting plays an important role for the conversion and storage of renewable energy sources. Here, we report a study on the electrocatalytic properties of the electrodeposited-film electrodes derived from irreversible electro-reduction/-oxidation of a molecular dinitrosyl iron complex (DNIC) {Fe(NO)2}9 [(Me6tren)Fe(NO)2]+ (Me6tren = tris[2-(dimethylamino)ethyl]amine) for the hydrogen evolution reaction (HER) and the oxygen evolution reaction (OER) in alkaline solution, individually. For HER, the overpotential and Tafel slope for the electrodeposited-film cathode are lower than those of the equiv.-weight Pt/C electrode. The electrodeposited-film anode for the OER is stable for 139 h. Integration of the electrodeposited-film cathode and anode into a single electrode-pair device for electrocatalytic water splitting exhibits an onset voltage of 1.77 V, achieving a geometrical current density of 10 mA cm−2. [ABSTRACT FROM AUTHOR]

This study was conducted to confirm that 1-site and 4-site ppU6-GGTA1-gRNA CRISPR vectors together with the pCX-Flag2-NLS1-Cas9-NLS2plasmid can both generate KO pigs by direct pronuclear microinjection. In total, 41 and 53 fertilized eggs were microinjected on 1-site and 4-site strategies, respectively. The 1-site construction generated a litter of 8 piglets, and 2 were mono-allelic mutant (mMt). The injection of 4-site constructions resulted in one biallelic mutant (bMt) and one mMt piglet in a litter of 7. Those 3 mMt pigs had a 4 bp deletion, 5 bp insertion, or 7 bp insertion at site I, and the bMt pig had 5 types of mutations at cleavage sites I and III. The expression of alpha-Gal on the bMt peripheral blood mononuclear cells (PBMCs) was reduced, and survival rate of bMt PBMCs was maintained as indicated by results of cultivation with sera of humans or Formosan Macaques. We concluded that mutant pigs could be generated by direct pronuclear microinjection of ppU6-GGTA1-gRNA CRISPR vectors with the pCX-Flag2-NLS1-Cas9-NLS2plasmid and that the 4-site strategy has a better mutant efficiency. Porcine U6 promoter was firstly used to express KO vectors and effectively generate mutant pigs, worthily to adopt for future KO studies. [ABSTRACT FROM AUTHOR]

We developed nonintegrated methods to reprogram mouse embryonic fibroblast (MEF) cells into induced pluripotent stem cells (iPSCs) using pig pOct4, pSox2, and pc-Myc as well as human hKLF4, hAID, and hTDG that were carried by plasmid vectors. The 4F method employed pOct4, pSox2, pc-Myc, and hKLF4 to derive iPSC clones with naive embryonic stem cell (ESC)-like morphology. These 4F clones expressed endogenous mouse Nanog protein and could generate chimeras. In addition to the four conventional reprogramming factors used in the 4F method, hAID and hTDG were utilized in a 6F method to increase the conversion efficiency of reprogramming by approximately five-fold. One of the 6F plasmid derived iPSC (piPSC) clones was shown to be germline transmission competent. [ABSTRACT FROM AUTHOR]

The rabbit is a useful animal model for regenerative medicine. We previously developed pluripotent rabbit embryonic stem cell (rbESC) lines using fresh embryos. We also successfully cryopreserved rabbit embryos by vitrification. In the present work, we combined these two technologies to derive rbESCs using vitrified-thawed (V/T) embryos. We demonstrate that V/T blastocysts (BLs) can be used to derive pluripotent rbESCs with efficiencies comparable to those using fresh BLs. These ESCs are undistinguishable from the ones derived from fresh embryos. We tested the developmental capacity of rbESCs derived from V/T embryos by BL injection experiments and produced chimeric kits. Our work adds cryopreservation to the toolbox of rabbit stem cell research and applications and will greatly expand the available research materials for regenerative medicine in a clinically relevant animal model. [ABSTRACT FROM AUTHOR]

A positive myocardial inotropic effect achieved using HNO/NO−, compared with NO⋅, triggered attempts to explore novel nitroxyl donors for use in clinical applications in vascular and myocardial pharmacology. To develop M-NO complexes for nitroxyl chemistry and biology, modulation of direct nitroxyl-transfer reactivity of dinitrosyl iron complexes (DNICs) is investigated in this study using a FeIII-porphyrin complex and proteins as a specific probe. Stable dinuclear {Fe(NO)2}9 DNIC [Fe(μ-MePyr)(NO)2]2 was discovered as a potent nitroxyl donor for nitroxylation of FeIII-heme centers through an associative mechanism. Beyond the efficient nitroxyl transfer, transformation of DNICs into a chemical biology probe for nitroxyl and for pharmaceutical applications demands further efforts using in vitro/in vivo studies. [ABSTRACT FROM AUTHOR]

We attempted to isolate ES cell lines using inner cell masses from high-quality cloned porcine blastocysts. After being seeded onto feeders, embryos had better (P < 0.05) attachment, outgrowth formation and primary colonization in both 2× and 3× aggregated cloned embryos (62.8, 42.6 and12.8% vs. 76.2, 55.2 and 26.2%, respectively) compared to the non-aggregated group (41.6, 23.4 and 3.9%). Effects of feeder types (STO vs. MEF) and serum sources (FBS vs. KSR) on extraction of cloned embryo-derived porcine ES cells were examined. More (17.1%) ntES cell lines over Passage 3 were generated in the MEF/KSR group. However, ntES cells cultured in KSR-supplemented medium had a low proliferation rate with defective morphology, and eventually underwent differentiation or apoptosis subsequently. Approximately 26.1, 22.7 and 35.7% of primary colonies were formed after plating embryos in DMEM, DMEM/F12 and α-MEM media, respectively. Survival rates of ntES cells cultured in α-MEM, DMEM and DMEM/F12 were 16.7, 4.3 and 6.8%, respectively (P > 0.05). We further examined the beneficial effect of TSA treatment of 3× aggregated cloned embryos on establishment of ntES cell lines. Primary colony numbers and survival rates of ntES cells beyond passage 3 were higher (P < 0.05) in those derived from TSA-treated 3× blastocysts (36.7 and 26.7%) than from the non-treated aggregated group (23.1 and 11.5%). These cells, remaining undifferentiated over 25 passages, had alkaline phosphatase activity and expressed ES specific markers Oct4, Nanog, Sox2, and Rex01. Moreover, these ntES cells successfully differentiated into embryoid bodies (EBs) that expressed specific genes of all three germ layers after being cultured in LIF-free medium. In conclusion, we have successfully derived putative porcine ntES cells with high efficiency from quality cloned embryos produced by embryo aggregation, and optimized the ES cell culture system suitable for establishing and maintaining ntES cell lines in undifferentiated state. [ABSTRACT FROM AUTHOR]

The monomeric {Fe(NO)2}10 dinitrosyliron complexes (DNICs) [K-18-crown-6-ether][(NO)2Fe(NC9H6-NH)] ( 1), [K-18-crown-6-ether][(NO)2Fe(NC9H6-O)] ( 2), and [K-18-crown-6-ether][(NO)2Fe(NC9H6-S)] ( 3) containing [N,NH], [N,O], and [N,S] ligations were synthesized by displacing the N, N, N′, N′-tetramethylethylenediamine (tmeda) group of {Fe(NO)2}10 (tmeda)Fe(NO)2 with quinolin-8-amide, quinolin-8-olate, and quinolin-8-thiolate, respectively. Anionic {Fe(NO)2}10 DNICs 1- 3 were characterized by IR, 1H NMR, and UV/Vis spectroscopy and single-crystal X-ray diffraction. The binding affinity of the ligands (i.e., [NC9H6NH]-, [NC9H6O]-, and [NC9H6S]-) toward the {Fe(NO)2}10 fragment was also investigated, and ligand substitution was found to follow the order [NC9H6S]- > [NC9H6O]- > [NC9H6NH]-. Displacement of the tmeda group of (tmeda)Fe(NO)2 by [K-18-crown-6-ether][SC6H4- o-NH2] and [PPh4][SC6H4- o-NH2] afforded monomeric {Fe(NO)2}10 DNIC [K-18-crown-6-ether][(NO)2Fe(SC6H4- o-NH2)] ( 4) and dimeric {Fe(NO)2}10-{Fe(NO)2}10 DNIC [PPh4]2[(NO)2Fe(μ-SC6H4- o-NH2)]2 ( 5), respectively. The function of the countercation in the formation of monomeric {Fe(NO)2}10 DNIC 4 or dimeric {Fe(NO)2}10-{Fe(NO)2}10 DNIC 5 was also demonstrated upon reduction of the {Fe(NO)2}9 DNICs [cation][(NO)2Fe(SC6H4- o-NH2)2] ( 6) {cation = K-18-crown-6-ether ( 6a), [PPh4] ( 6b)}. In contrast to the formation of thiolate-containing {Fe(NO)2}9 DNICs through nucleophilic cleavage of the bridging thiolates of Roussin's red esters, the transformation of complexes 4 and 5 into complexes 6a and 6b, respectively, in the presence of disulfide provides a new synthetic pathway for the synthesis of thiolate-containing {Fe(NO)2}9 DNICs. [ABSTRACT FROM AUTHOR]

A Ni(II)-mixed-chalcogenolate carbonyl complex, [PPN][Ni(CO)(SePh)2(S- p-C6H4-Cl)] ( 1) was obtained from the treatment of [CpNi(μ-S- p-C6H4-Cl)]2 and fac-[PPN][Fe(CO)3(SePh)3]. Using 1 as a precursor, two four-coordinated square planar Ni(II)-chalcogenolate/chalcogenol complexes, [PPN][Ni-(SePh)(N(CH2CH2S)2(CH2CH2SH))] ( 2) and [Ni(PEt3)(N(CH2CH2S)2(CH2CH2SH))] ( 4) containing the thiol in the secondary coordination sphere have been synthesized and identified by single-crystal X-ray diffraction. Analyses of structural data and vibrational energy (νSH = 2431 cm−1 for 4) reveal that the pendant thiol in complexes 2 and 4, respectively do not form intramolecular interaction with the nickel and thiolato sulfur. Compared to previously reported analogues coordinated by [P( o-C6H4S)2( o-C6H4SH)]2−ligand (Chen, C.-H.; Lee, G.-H.; Liaw, W.-F. Inorg. Chem. 2006, 45, 2307), the lack of intramolecular interaction in complexes 2 and 4 may be attributed to the rotatable C-S and C-C bonds of [N(CH2CH2S)2(CH2CH2SH)]2− ligand, precluding the pendant thiol from pointing a direction to the nickel and thiolato sulfur. [ABSTRACT FROM AUTHOR]

Abstract: The reaction of [Fe(μ-I)(NO)2]2 and TMEDA in a 1:2 molar ratio in THF affords the neutral five-coordinate DNIC [(TMEDA)Fe(NO)2I] (1). The single-crystal X-ray structure shows that the geometry of iron center of complex 1 is best described as a distorted trigonal bipyramidal with two nitrosyl groups positioned in the equatorial plane. The EPR spectrum of complex 1 displays the six-line signal with g =2.031 (a I =37.6G) at 298K. The coincident g values of EPR among complex 1, protein-bound DNICs and low-molecular-weight DNICs implicate that the five-coordinate DNICs may exist in biological system. The interconversion between complex 1 and [(TMEDA)Fe(NO)2] (2) reveals that the {Fe(NO)2}9 DNICs containing [amine, amine] ligation mode could be stabilized by the five-coordinated geometry while the {Fe(NO)2}10 DNICs containing [amine, amine] ligation mode favors the four coordination sphere. In addition, the transformation from complex 1 to [Fe(NO)2(C3H3N2)]4 (3), [Fe(μ-SPh)(NO)2]2 (4), [PPh4][(PhS)2Fe(NO)2] (5) and [Na-18-crown-6-ether][(C3H3N2)2Fe(NO)2] (6), respectively, in the presence of thiolates or imidazolates indicates that complex 1 could be employed as the precursor for the syntheses of the DNICs containing the [N,N]/[N,S]/[S,S] different ligations. [Copyright &y& Elsevier]

Mononuclear, distorted square planar [NiII(ER)(P(o-C6H4S)2(o-C6H4SH))]- (ER = SePh (1), 2-S-- C4H3S (2)) with a S-H proton directly interacting with both nickel and sulfur atoms were prepared by reaction of [Ni(CO)(SePh)3]/[Ni(CO)(2-S-C4H3S)3] and P(o-C6H4SH)3, individually. The presence of combinations of intramolecular [Ni-S…H-SR]/[Ni…H-SR] interactions was verified in the solid state by the observation of an IR vSH stretching band (2273 and 2283 cm-1 (KBr) for complexes 1 and 2, individually) and ¹H NMR spectra (δ 8.079 (d) (CD2Cl2) and 8.39 (d) (C4D8O) ppm (-SH) for complexes 1 and 2, respectively) and subsequently confirmed by X-ray diffraction study. The exo-thiol proton (o-C6H4SH) in complexes 1 and 2 was identified as a D2O exchangeable proton from NMR and IA studies and was quantitatively removed by Lewis base Et3N to yield Ni(II) dimer [NiII(P(o-C6H4S)3)]62- (5). Instead of the ligand-based oxidation to form dinuclear Ni(Il) complexes and dichalcogenide, oxidation of THF-CH3CN solution of complexes 1 and 2 by O2 resulted in the formation of the mononuclear, distorted trigonal bipyramidal [NiII(ER)(P(o-C6H4S)3)y (ER = SePh (3), 2-S-C4H3S (4)) accompanied by byproduct H2O identified by ¹H NMR, respectively. The 4.2 K EPA spectra of complexes 3 and 4 exhibiting high rhombicities with three principal g values of 2.304, 2.091, and 2.0 are consonant with Ni(Ill) with the odd electron in the d22 orbital. Complex 3 undergoes a reversible NiIII/II process at E… = -0.67 V vs Ag/AgCl in MeCN. [ABSTRACT FROM AUTHOR]

Mononuclear, distorted trigonal bipyramidal [PPN][NiIII(R)- (P(C6H3-3-SiMe3-2-S)3)] (R = Me (1); R = Et (2)) were prepared by reaction of [PPN][NiIIICI(P(C6H3-3-SiMe3-2-S)3)] and CH3MgCI/ C2H5MgCI, individually. EPR, SQUID studies as well as DFT computations reveal that the NiIII in 1 has a low-spin d7 electronic configuration in a distorted trigonal bipyramidal ligand field. The Ni-C bond lengths of 1.994(3) Å in 1 and 2.015(3) Å in 2 are comparable to that in the NiIII-methyl state of MCR (∼2.04 Å) (Sarangi, R.; Dey, M.; Ragsdale, S. W. Biochemistry 2009, 48, 3146). Under a CO atmosphere, CO triggers homolytic cleavage of the NiIIICH3 bond in 1 to produce NiIII-thiolate carbonyl [PPN][NiIII(CO)(P(C6H3-3-SiMe3-2-S)3)] (3). Additionally, protonation of 1 with phenylthiol generates NiIII-thiolate [PPN][NiIIISPh)- (P(C6H3-3-SiMe3-2-S)3)] (4). [ABSTRACT FROM AUTHOR]

This retrospective study attempts to elucidate the relevance of the interval between human chorionic gonadotropin priming and oocyte pick-up (hCG-OPU) to the euploidy probability of biopsied blastocysts in preimplantation genetic tests for aneuploidy (PGT-A) cycles. A total of 1889 blastocysts from 511 patients undergoing PGT- A cycles were used. An analysis of generalized estimating equations (GEE) was used to identify whether the hCG–OPU interval is associated with euploidy probabilities of blastocysts. Accordingly, maternal age (OR: 0.925, 95% CI: 0.903–0.948, p < 0.001) and the hCG–OPU interval (OR: 1.138, 95% CI: 1.028–1.260, p = 0.013) were the two significant factors associated with the euploidy probabilities. The Cochran-Armitage trend test demonstrated that the blastocyst euploidy percentage increased progressively with the increasing hCG-OPU interval in normal responders (p = 0.006) and advanced maternal age (age ≥38 years; p = 0.020) groups. In normal responders, the euploidy rate was highest in the 38–39 h interval (43.1%, 47/109). In contrast, the euploidy rate was lowest in the 34–35 h interval (28.7%, 29/105). In conclusion, the present study demonstrated that at an hCG-OPU interval between 34–39 h, the longer the hCG-OPU interval, the higher the probability of euploidy for blastocysts. [ABSTRACT FROM AUTHOR]

The present study aimed to establish embryonic stem (ES) cell lines, i.e., ntES cells, using rabbit blastocyst stage embryos cloned by somatic cell nuclear transfer. First, we investigated the development of cloned rabbit embryos reconstructed with normal fibroblasts and fibroblasts transfected with enhanced green fluorescence protein (eGFP). Blastocyst rates were 27.4% and 23.9%, respectively, for the embryos reconstructed with normal fibroblasts and fibroblasts transfected with eGFP compared with that from the parthenogenetic group (43.1%). One ntES cell line was established from embryos reconstructed with eGFP-transfected fibroblasts (1 of 17, 5.9%), and three ntES cell lines were derived from those with normal fibroblasts (3 of 17, 17.6%). All the ntES cell lines retained alkaline phosphatase activity and expressed ES cell–specific markers SSEA-4, Oct-4, TRA-1-60, and TRA-1-81. The pluripotency was further confirmed by reverse transcription–polymerase chain reaction analyses of Oct-4 , Nanog , and Sox-2 expressions in ntES cell lines. The differentiation capacity of ntES cells was also examined in vitro and in vivo , by which these ntES cell lines were able to differentiate into all three germ layers through embryoid bodies and teratomas. In conclusion, it is apparent that the efficiency of ntES cells derived using eGFP-transfected donor cells is lower than that with nontransfected, normal fibroblasts donor cells. Similar to those from parthenogenetic embryos, all ntES cell lines derived from cloned rabbit embryos are able to express pluripotency markers and retain their capability to differentiate into various cell lineages both in vitro and in vivo . [ABSTRACT FROM AUTHOR]

Abstract: The objectives of this study were to characterize the nuclear and cytoskeletal changes of pig oocytes during in vitro maturation (IVM) and the development of the reconstructed embryos after injection with membrane intact or disintegrated donor cells. Cumulus-oocyte complexes (COCs) were collected from abattoir ovaries by follicle (2–8mm) aspiration. In Experiment 1, COCs were cultured in NCSU-23 medium for 0, 11, 22, 33, and 44h. Oocytes were fixed at different time points for nuclear and cytoskeletal labeling. Forty-three percent and 75% oocytes progressed to MII stage at 33 and 44h after IVM culture, respectively. Dynamic shift of spindle and cytoplasmic microtubules was evident. In Experiment 2, matured oocytes were injected with either the whole cumulus cell with or without intact cell membranes after enucleation. The reconstructed oocytes were fixed at 0, 2, or 4h after cell injection for nuclear and cytoskeletal evaluation. When an intact cumulus cell was injected, the injected cell remained intact within 4h after injection. When a cell with disintegrated membrane was injected, 59–63% (n =146) of the injected cell underwent premature chromosome condensation (PCC). In Experiment 3, the reconstructed pig oocytes received membrane-disintegrated cumulus cells or fetal fibroblasts were cultured in PZM medium. The blastocyst rate of the fibroblast-injected embryos was 10%, which was lower than the non-cloned parthenotes (33%, P <0.05) but higher than the cumulus cell-injected embryos (2.7%). These results suggest that pig oocytes are subjected to nuclear and cytoskeletal reorganization during maturation. Pig oocytes injected with membrane-disintegrated fibroblast cells support better blastocyst development of the cloned embryos. [Copyright &y& Elsevier]

Reaction of compounds [(NO)2Fe(TMEDA)] and [BH4]− affords the first hydride-containing dinitrosyl iron complex (DNIC) [(NO)2Fe(η2-BH4)]− (2). Hydride-insertion reactivity of DNIC 2 promotes the reductive transformation of CS2 into DNIC [(NO)2Fe(η3-HCS2)]−, the first DNIC featuring both linear/bent NO ligands and Fe 3dz2-to-HCS2π* backbonding interaction. [ABSTRACT FROM AUTHOR]