138 FATTY ACID β-OXIDATION REGULATES BOVINE EMBRYO OXYGEN METABOLISM

2013 ◽  
Vol 25 (1) ◽  
pp. 216
Author(s):  
P. J. McKeegan ◽  
F. G. Guerif ◽  
R. G. Sturmey
Keyword(s):  
Oxygen Consumption ◽  
Fatty Acid ◽  
Atp Synthesis ◽  
Oxygen Metabolism ◽  
Blastocyst Stage ◽  
Early Embryo ◽  
Metabolic Energy ◽  
Acid Oxidation ◽  
Bovine Embryo

Mammalian oocytes are rich in endogenous lipid, which provides a potential source of metabolic energy during oocyte maturation and early embryo development after fertilisation. Despite this, studies on early embryo metabolism have focussed on consumption of substrates from the culture medium, with comparatively little consideration for endogenous stores. Additionally, the interaction between fatty acid β-oxidation and oxygen consumption during pre-implantation development has not been investigated. This study has investigated the relationship between embryo fatty acid metabolism and oxygen consumption rate (OCR), a marker of overall oxidative metabolic activity. The amount of oxygen consumption by bovine blastocysts coupled to ATP synthesis was assessed by measuring embryo OCR following acute treatment with oligomycin, which inhibits ATP synthase. Bovine embryos were then produced in vitro and cultured from zygote to blastocyst stage (8 days) in the presence of β-mercaptoacetate (BMA), a competitive inhibitor of fatty acid β-oxidation, or L-carnitine, a cofactor that promotes β-oxidation. OCR of individual embryos was measured using a noninvasive, highly sensitive oxygen probe (Unisense), whereas OCR of embryos in groups was determined using the Oxygen Biosensor (OBS) fluorimetric assay (BD Biosciences, Erembodegem, Belgium). Mean OCR measured by each technique correlated well. Data are presented as means ± SEM. The effect of acute inhibition were compared by a paired t-test, with pre-treated embryos acting as their own control, whereas the effect of L-carnitine and BMA on OCR was compared across treatment groups using ANOVA with Dunn’s test for unequal groups post hoc, using Sigmaplot (Systat Software Inc., San Jose, CA, USA). Treatment with oligomycin caused blastocyst OCR to fall from 23.5 ± 3.3 to 7.9 ± 2.5 pmol/embryo/h (n = 3; P = 0.03). Inhibition of β-oxidation by BMA led to a modest, but consistent increased OCR in cleavage stage embryos: 4-cell: 14.8 ± 7.2 (n = 12) v. 8.9 ± 1.1 pmol/embryo/h (n = 3) control; 8-cell: 15.8 ± 2.2 (n = 6) v. 11.9 ± 2.3 pmol/embryo/h (n = 3) control. At the morula stage, BMA led to a significant rise in OCR (26.8 ± 3.6 (n = 7) v. 7.8 ± 1.7 pmol/embryo/h (n = 3) control; P = 0.003) and at the blastocyst stage, OCR was significantly elevated in the presence of BMA; 41.3 ± 2.6 (n = 19) v. 16.9 ± 1.3 pmol/embryo/h (n = 14) control (P = 0.01). By contrast, when groups of embryos were cultured in the presence of L-carnitine, blastocyst OCR was reduced by 45% from 24.7 ± 5.1 (group control) to 15.9 ± 3.7 pmol/embryo/h (n = 30; P < 0.06). We report that 66.5% of oxygen consumption by bovine blastocysts is oligomycin sensitive, meaning that 33.5% of oxygen consumption is used for processes other than ATP synthesis. Surprisingly, inhibition of fatty acid β-oxidation caused total bovine embryo OCR to rise, although the reasons for this are unclear. It is possible that inhibiting β-oxidation leads to an accumulation of free fatty acids in the mitochondrion, which can disrupt the inner membrane and lead decoupling of ATP synthesis from oxygen consumption. Thus, manipulating fatty acid oxidation in the early embryo dysregulates oxidative metabolism; the implications of this are under further investigation.

123 CHANGES IN EXPRESSION OF GENES ASSOCIATED WITH GENETIC VARIATION IN PRE-IMPLANTATION DEVELOPMENT OF THE BOVINE EMBRYO

2014 ◽  
Vol 26 (1) ◽  
pp. 175
Author(s):  
M. S. Ortega ◽  
J. B. Cole ◽  
T. S. Sonstegard ◽  
P. J. Hansen
Keyword(s):  
Genetic Variation ◽  
Intracellular Transport ◽  
Blastocyst Stage ◽  
Acid Oxidation ◽  
Lethal Gene ◽  
Bovine Embryo ◽  
Cell Stage ◽  
Glycoprotein Synthesis ◽  
Stage Of Development

The objective was to identify patterns of expression during the pre-implantation period of several genes associated with genetic variation in fertility (CWC15) or development to the blastocyst stage (C1QB, MON1B, PARM1, PCCB, PMM2, TBC1D24, and WBP1). These genes are involved in cellular processes such as mRNA splicing, immune protection, fatty acid oxidation, resistance to apoptosis, glycoprotein synthesis, and intracellular transport. Embryos were produced in vitro from slaughterhouse oocytes and semen using a mix of Bos taurus and Bos indicus cows and bulls. Pools of 40 matured oocytes or embryos at the 2-cell [27–31 h post-insemination (hpi)], 3- to 4-cell (46–52 hpi), 5- to 8-cell (49–59 hpi), 9- to 16-cell (72–75 hpi), morula (120–123 hpi), and blastocyst (168–171 hpi) stages were collected. The RNA was purified and synthesised into cDNA for real-time qPCR analysis. The YWHAZ, GAPDH, and SDHA were used as steady-state controls of expression. A total of 5 pools were analysed for each of the 6 stages. The C1QB was not detected at any stage; however, transcript amounts for the other genes were affected by stage of development (P < 0.05). The WBP1 remained low from the oocyte to the 5- to 8-cell stage (fold-change relative to matured oocytes: 1.0 ± 0.2 v. 1.4 ± 0.2), increased at the 9- to 16-cell stage (14.8 ± 0.2), and decreased to the blastocyst stage (7.1 ± 0.2). The expression pattern of PARM1 was similar, with greatest expression at the 9- to 16-cell stage. In contrast, expression of PMM2 and TBC1D24 was highest at the 2-cell stage and decreased at the morula and blastocyst stages. Expression of CWC15, MON1B, and PCCB decreased steadily from the oocyte to the blastocyst stage. Given that the major round of embryonic genome activation occurs at the 8- to 16-cell stage, it is possible that PARM1 and WBP1 play important roles around this time. The PMM2 and TBC1D24 may represent genes activated before the 8- to 16-cell stage. The CWC15 has been identified as a lethal gene; results suggest lethality occurs after the blastocyst stage. Further research will clarify the role and importance of these genes in the early development of the bovine embryo. The authors acknowledge support from AFRI Grant No. 2013–68004–20365 from USDA NIFA.

137 ROLE OF GLUCOSE AND FATTY ACID METABOLISM IN PORCINE EARLY EMBRYO DEVELOPMENT

2008 ◽  
Vol 20 (1) ◽  
pp. 149 ◽  
Author(s):  
R. G. Sturmey ◽  
H. J. Leese
Keyword(s):  
Energy Source ◽  
Fatty Acid ◽  
Glucose Metabolism ◽  
Embryo Development ◽  
Alternative Energy ◽  
Tca Cycle ◽  
Early Embryo ◽  
Metabolic Energy ◽  
The Tca Cycle

Glucose metabolism plays an important role in the preimplantation development of porcine embryos in vitro. As in mammalian species generally, a proportion of glucose consumed is converted to lactate by aerobic glycolysis generating small amounts of ATP, with the remainder oxidized by the TCA cycle. However, a striking feature of the porcine early embryo is the large amount of lipid present as triglyceride (TG), which represents an alternative energy source. The TG is metabolized via β-oxidation, producing acetyl Co A, which in turn is oxidized by the TCA cycle. This sequence of reactions requires a constant supply of carbohydrate to provide oxaloacetate (OA) to prime the TCA cycle. The provision of OA from pyruvate arising from glycolysis may represent an alternative role for glucose in early pig embryo development. We have therefore sought to determine the importance of interplay between glucose and TG metabolism in porcine embryos in vitro. Porcine embryos were generated in vitro by fertilization of in vitro-matured oocytes collected from abattoir-derived ovaries. Oocytes were matured in defined maturation medium and embryos cultured in NCSU23. Glucose consumption, lactate production, and TG content of single porcine blastocysts cultured throughout development in the presence of methyl palmoxirate (MP), an inhibitor of TG metabolism, were measured as described by Sturmey RG and Leese HJ 2003 (Reprod. 126, 197–204). The capacity of zygotes to form blastocysts when cultured with OA in place of glucose in the presence or absence of MP and the amount of TG in blastocysts grown in either glucose or OA-containing medium were then determined (6 replicates). When TG metabolism was inhibited, porcine blastocysts consumed significantly more glucose (32 � 9 pmol/embryo/h v. 11 � 1 pmol/embryo/h; P < 0.05; n = 34) and produced higher amounts of lactate (35 � 4 pmol/embryo/h v. 10 � 0.8 pmol/embryo/h; P < 0.01; n = 34). Blastocyst rates did not differ significantly between embryos grown in the presence of glucose or OA, and in blastocysts grown in OA-NCSU the TG content was significantly reduced (155 � 8 ng v. 240 � 12 ng; P = 0.015; n = 41). All embryos cultured in OA-containing medium in the presence of MP failed to develop beyond the zygote stage. The data support the notion that porcine embryos can use endogenous TG as a metabolic energy source. When this is prevented by chemical inhibition, the embryo upregulates glycolysis and glucose oxidation as an alternate means of generating ATP. When cultured in medium containing OA, a compound that cannot generate ATP per se, embryo development was similar to controls, again suggesting the ability to use endogenous energy stores, a proposition reinforced by a significant fall in the levels of TG in the presence of OA. However, by inhibiting β-oxidation in the absence of glucose, porcine embryos were unable to develop. The relationship between TG and glucose metabolism by porcine embryos is analogous to the glucose/fatty acid cycle in whole animals where glucose and TG can be used as energy sources, but in a reciprocal manner. The data also demonstrate the plasticity of energy metabolism by porcine early embryos.

145 CHANGES IN OXYGEN COMSUMPTION RATES OF EMBRYOS IN KOREAN CATTLE

2010 ◽  
Vol 22 (1) ◽  
pp. 231
Author(s):  
C. Y. Choe ◽  
S. R. Cho ◽  
J. K. Son ◽  
S. H. Choi ◽  
C. Y. Cho ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Embryo Quality ◽  
Blastocyst Stage ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Cell Stage ◽  
Significant Difference ◽  
Morula Stage

Oxygen consumption has been regarded as a useful indicator for assessment of mammalian embryo quality. This study was carried out to identify whether oxygen consumption rates measured in bovine embryos using SECM can be used as a standard criteria to evaluate bovine embryo quality. Oxygen consumption of bovine embryos at various developmental stages was measured and analyzed using SECM and ANOVA analysis, respectively. We found that the oxygen consumption significantly increased in blastocyst-stage embryos compared to other stage embryos (from 2-cell stage to morula stage), indicating that oxygen consumption reflects the cell number (5.2-7.6 × 1014 mol-1 s-1 v. 1.2-2.4 × 1014 mol-1 s-1, P < 0.05). There was no significant difference between 2-cell-stage embryos and 8-cell-stage embryos. In the morula-stage embryos, the oxygen consumption of in vivo derived embryos was significantly higher than that of in vitro produced embryos (4.0 × 1014 mol-1 s-1 v. 2.4 × 1014 mol-1 s-1, P < 0.05). However, there was no significant difference in consumption of oxygen by in vivo and in vitro derived bovine blastocyst-stage embryos (P > 0.05). Good-quality embryos with grade 1 or 2 showed significantly higher oxygen consumption than grade 3 or 4 embryos. These results showed that SECM could measure oxygen consumption in bovine embryos and the oxygen consumption could reflect embryonic development stage and embryo quality.

scholarly journals The PPARδ Agonist GW501516 Improves Lipolytic/Lipogenic Balance through CPT1 and PEPCK during the Development of Pre-Implantation Bovine Embryos

2019 ◽  
Vol 20 (23) ◽  
pp. 6066 ◽  
Author(s):  
Muhammad Idrees ◽  
Lianguang Xu ◽  
Marwa El Sheikh ◽  
Tabinda Sidrat ◽  
Seok-Hwan Song ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Embryo Development ◽  
Embryo Quality ◽  
Acid Oxidation ◽  
Peroxisome Proliferator ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Peroxisome Proliferator Activated Receptors

The PPARs (peroxisome proliferator-activated receptors) play critical roles in the regulation of lipid and glucose metabolism. PPARδ, a member of the PPARs family, is associated with decreased susceptibility to ectopic lipid deposition and is implicated in the regulation of mitochondrial processes. The current study aimed to determine the role of PPARδ in fatty acid β-oxidation and its influence on PEPCK for the lipogenic/lipolytic balance during in vitro bovine oocyte maturation and embryo development. Activation of PPARδ by GW501516, but not 2-BP, was indicated by intact embryonic PEPCK (cytosolic) and CPT1 expression and the balance between free fatty acids and mitochondrial β-oxidation that reduced ROS and inhibited p-NF-κB nuclear localization. Genes involved in lipolysis, fatty acid oxidation, and apoptosis showed significant differences after the GW501516 treatment relative to the control- and 2-BP-treated embryos. GSK3787 reversed the PPARδ-induced effects by reducing PEPCK and CPT1 expression and the mitochondrial membrane potential, revealing the importance of PPARδ/PEPCK and PPARδ/CPT1 for controlling lipolysis during embryo development. In conclusion, GW501516-activated PPARδ maintained the correlation between lipolysis and lipogenesis by enhancing PEPCK and CPT1 to improve bovine embryo quality.

TetR-family transcriptional repressor Thermus thermophilus FadR controls fatty acid degradation

Microbiology ◽  
2011 ◽  
Vol 157 (6) ◽  
pp. 1589-1601 ◽  
Author(s):  
Yoshihiro Agari ◽  
Kazuko Agari ◽  
Keiko Sakamoto ◽  
Seiki Kuramitsu ◽  
Akeo Shinkai
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Target Genes ◽  
Thermus Thermophilus ◽  
Acyl Chain ◽  
Metabolic Energy ◽  
Straight Chain ◽  
Fatty Acid Degradation ◽  
Acid Degradation

In the extremely thermophilic bacterium Thermus thermophilus HB8, one of the four TetR-family transcriptional regulators, which we named T. thermophilus FadR, negatively regulated the expression of several genes, including those involved in fatty acid degradation, both in vivo and in vitro. T. thermophilus FadR repressed the expression of the target genes by binding pseudopalindromic sequences covering the predicted −10 hexamers of their promoters, and medium-to-long straight-chain (C10–18) fatty acyl-CoA molecules were effective for transcriptional derepression. An X-ray crystal structure analysis revealed that T. thermophilus FadR bound one lauroyl (C12)-CoA molecule per FadR monomer, with its acyl chain moiety in the centre of the FadR molecule, enclosed within a tunnel-like substrate-binding pocket surrounded by hydrophobic residues, and the CoA moiety interacting with basic residues on the protein surface. The growth of T. thermophilus HB8, with palmitic acid as the sole carbon source, increased the expression of FadR-regulated genes. These results indicate that in T. thermophilus HB8, medium-to-long straight-chain fatty acids can be used for metabolic energy under the control of FadR, although the major fatty acids found in this strain are iso- and anteiso-branched-chain (C15 and 17) fatty acids.

scholarly journals Interleukin-6 Increases Insulin-Stimulated Glucose Disposal in Humans and Glucose Uptake and Fatty Acid Oxidation In Vitro via AMP-Activated Protein Kinase

Diabetes ◽  
2006 ◽  
Vol 55 (10) ◽  
pp. 2688-2697 ◽  
Author(s):  
A. L. Carey ◽  
G. R. Steinberg ◽  
S. L. Macaulay ◽  
W. G. Thomas ◽  
A. G. Holmes ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Protein Kinase ◽  
Glucose Uptake ◽  
Fatty Acid Oxidation ◽  
Interleukin 6 ◽  
Glucose Disposal ◽  
Acid Oxidation ◽  
Amp Activated Protein Kinase

P–219 mtDNA content in bovine cumulus cells does not predict oocytés developmental competence

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Á Martíne. Moro ◽  
I Lamas-Toranzo ◽  
L González-Brusi ◽  
A Pérez-Gómez ◽  
P Bermejo-Álvarez
Keyword(s):  
Cumulus Cell ◽  
Cumulus Cells ◽  
Oocyte Quality ◽  
Blastocyst Stage ◽  
Early Embryo ◽  
Developmental Competence ◽  
Success Rates ◽  
Developmental Potential ◽  
Mtdna Sequence

Abstract Study question Does cumulus cell mtDNA content correlate with oocyte developmental potential in the bovine model? Summary answer The relative amount of mtDNA content did not vary significantly in oocytes showing different developmental outcomes following IVF What is known already Cumulus cells are closely connected to the oocyte through transzonal projections, serving essential metabolic functions during folliculogenesis. These oocyte-supporting cells are removed and discarded prior to ICSI, thereby constituting an interesting biological material on which to perform molecular analysis aimed to predict oocyte developmental competence. Previous studies have positively associated oocytés mtDNA content with developmental potential in both animal models and women. However, it remains debatable whether mtDNA content in cumulus cells could be used as a proxy to infer oocyte developmental potential. Study design, size, duration Bovine cumulus cells were allocated into three groups according to the developmental potential of the oocyte: 1) oocytes developing to blastocysts following IVF (Bl+Cl+), 2) oocytes cleaving following IVF but arresting their development prior to the blastocyst stage (Bl-Cl+), and 3) oocytes not cleaving following IVF (Bl-Cl-). Relative mtDNA content was analysed in 40 samples/group, each composed by the cumulus cells from one cumulus-oocyte complex (COC). Participants/materials, setting, methods Bovine cumulus-oocyte complexes were obtained from slaughtered cattle and individually matured in vitro (IVM). Following IVM, cumulus cells were removed by hyaluronidase treatment, pelleted, snap frozen in liquid nitrogen and stored at –80 ºC until analysis. Cumulus-free oocytes were fertilized and cultured in vitro individually and development was recorded for each oocyte. Relative mtDNA abundance was determined by qPCR, amplifying a mtDNA sequence (COX1) and a chromosomal sequence (PPIA). Statistical differences were tested by ANOVA. Main results and the role of chance Relative mtDNA abundance did not differ significantly (ANOVA p &gt; 0.05) between the three groups exhibiting different developmental potential (1±0.06 vs. 1.19±0.05 vs. 1.11±0.05, for Bl+Cl+ vs. Bl-Cl+ vs. Bl-Cl-, mean±s.e.m.). Limitations, reasons for caution Experiments were conducted in the bovine model. Although bovine folliculogenesis, monoovulatory ovulation and early embryo development exhibit considerable similarities with that of humans, caution should be taken when extrapolating these data to humans. Wider implications of the findings: The use of molecular markers for oocyte developmental potential in cumulus cells could be used to enhance success rates following single-embryo transfer. Unfortunately, mtDNA in cumulus cells was not found to be a good proxy for oocyte quality. Trial registration number Not applicable

Metabolic influences on satiety in rats receiving parenteral nutrition

1994 ◽  
Vol 266 (2) ◽  
pp. R381-R386 ◽  
Author(s):  
J. L. Beverly ◽  
M. M. Meguid ◽  
Z. J. Yang ◽  
M. X. Yue ◽  
B. L. Fetterman
Keyword(s):  
Fatty Acid ◽  
Food Intake ◽  
Parenteral Nutrition ◽  
Metabolic Energy ◽  
Acid Oxidation ◽  
Relative Importance ◽  
Phase 2 ◽  
Light Phase ◽  
Single Intraperitoneal Injection ◽  
And Control

Food intake is reduced during parenteral nutrition (PN) proportionally to the amount of calories or composition of the solution infused. The relative importance of infused glucose and lipid, 50 and 30% of PN kilocalories, respectively, in reducing food intake during PN was examined. Glycolysis, fatty acid oxidation, or both were acutely disrupted with 2-deoxy-D-glucose (2-DG) and mercaptoacetate (MA). Rats receiving intravenous infusions of saline or a PN solution providing 100% of total daily calories (PN-100) received a single intraperitoneal injection of saline, 2-DG, and/or MA during the early light phase. 2-DG (1.4 or 2.2 mmol/kg) did not initiate feeding in PN-100 rats, although hyperglycemia was evident in all rats 1 h after 2-DG. Food intake of PN-100 rats after MA (0.4 mmol/kg) was approximately 50% that of control rats. When 2-DG (1.4 mmol/kg) and MA (0.4 mmol/kg) were administered concomitantly, PN-100 and control rats consumed the same amount of food. During PN-100, rats appeared to be more sensitive to losing metabolic energy derived from lipid than from glucose.

scholarly journals 136 EVIDENCE OF A DIRECT EFFECT OF P4 ON IVF-DERIVED BOVINE 8-CELL EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 219 ◽  
Author(s):  
C.E. Ferguson ◽  
T.R. Davidson ◽  
M.R.B. Mello ◽  
A.S. Lima ◽  
D.J. Kesler ◽  
...  
Keyword(s):  
Embryo Development ◽  
Direct Effect ◽  
Blastocyst Stage ◽  
Control Group ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Direct Role ◽  
Treatment Groups ◽  
And Control

There has been much debate over a direct role for progesterone (P4) in early bovine embryo development. While previous attempts to supplement bovine embryos in vitro with P4 produced results that vary and are often contradictory, this may be a response of administering P4 at inappropriate times. Therefore, the objective of these experiments was to determine if P4 could exert a direct effect on developing IVF-derived bovine embryos when administered at an appropriate time of embryo development. In Exp. I, IVF-derived bovine 8-cell embryos were randomly allotted to treatments: (1) control, CR1aa medium (n = 168); (2) vehicle, CR1aa + ETOH (0.01%) (n = 170); and (3) P4, CR1aa + ETOH + P4 (20 ng/mL in 50-μL droplet) (n = 173). In Exp. II, IVF-derived bovine 8-cell embryos were randomly allotted to treatments: (1) control, CR1aa medium (n = 160); (2) vehicle, CR1aa + DMSO (0.01%) (n = 180); and (3) P4, CR1aa + DMSO (0.01%) + P4 (20 ng/mL in 50-μL droplet) (n = 170). All embryos were evaluated on Days 6 to 9 post-insemination and rates calculated from 8-cell embryos. In Exp. I, ETOH tended to have a detrimental effect with significantly fewer (P < 0.05) embryos (53%) developing to the blastocyst stage on Day 7 compared with the control (62%) and P4 (71%) groups. At Day 7, significantly more embryos cultured in P4 (71%) developed to the blastocyst stage compared with the control group (62%). P4 treatment significantly increased the number of Grade 1 blastocysts (25%) on Day 7 compared with vehicle (15%) and control (17%) groups. At the end of culture, there were also significantly more Day 9 hatched blastocysts in the P4 group (33%) compared with vehicle (22%) and control (21%) groups. Supplementing P4 in the culture medium increased the rate of development, resulting in significantly more blastocysts (8%) on Day 6 and hatched blastocysts (21%) on Day 8 compared with vehicle (3% and 12%) and control (0% and 8%) groups, respectively. In Exp. II, there were no significant differences between treatment groups for Day 7 blastocysts (control 54%, DMSO 61%, P4 57%) and Day 9 hatched blastocysts (control 46%, DMSO 51%, P4 46%). However, there were significantly more Grade 1 blastocysts in the P4 group (22% and 36%) on Days 6 and 8 compared with vehicle (11% and 23%) and control (13% and 23%) groups, respectively. The lack of improvement in Day 7 blastocysts and Day 9 hatched blastocysts rates leads to further uncertainty in understanding the P4 vehicle interactions. In conclusion, the results of these two experiments indicate that P4 can exert a direct effect on the developing IVF-derived bovine embryo; however, due to P4 vehicle interactions; other inert vehicles need to be explored to further evaluate the direct effects of P4 on the developing bovine embryo.

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