In vitro activation of primordial follicles could serve as a safe method to preserve fertility in patients with cancer subjected to ovarian tissue cryopreservation during oncotherapy, however the culture medium for this purpose requires to be optimized. Granulosa cell conditioned medium (GCCM) has been recognized to enhance primordial follicle activation and the present study was conducted to understand whether addition of pyruvate, a combination of insulin, transferrin and selenium (ITS) or testosterone to GCCM could improve its efficiency in this regard. To this end, 1-day-old mouse ovaries were cultured in four different media including CON (control; containing GGCM only), PYR (containing GCCM plus pyruvate), ITS (containing GCCM plus ITS) or TES (containing GCCM plus testosterone) for 11 days. Furthermore, follicular dynamics and gene expression of factors involved in follicular development were assessed using histological examination and RT-PCR, respectively, on days 5 and 11 of culture. Pyruvate decreased follicular activation, but it enhanced the progression of follicles to the primary stage. Moreover, it upregulated Bmp15 and Cx37 (P < 0.05). In the ITS group, activation of follicles was not affected and total number of follicles was reduced by day 11 of culture. Additionally, ITS downregulated Pi3k, Gdf9, Bmp15 and Cx37 (P < 0.05). Although testosterone did not affect primordial follicle activation, it enhanced the development of follicles up to the preantral stage (P < 0.05). Furthermore, testosterone inhibited the expression of Pten but stimulated the expression of Gdf9 and Cx37 (P < 0.05). In conclusion, the present study revealed that inclusion of pyruvate and testosterone into GCCM could enhance the early development of follicles in cultured 1-day-old mouse ovaries.

Chlorimuron antagonized the activity of quizalofop on broadleaf signalgrass when applied in greenhouse studies as a postemergence tank mix. In vitro leaf disc assays utilizing 14C-acetate or 14C-pyruvate as substrates were conducted to ascertain the effect of clorimuron and quizalofop on fatty acid biosynthesis and to determine if antagonism between the two herbicides occurs at the biochemical sites of action. Incorporation of 14C-acetate in control treatments was linear with time to 120 min. Acetate incorporation in the presence of quizalofop (1.1 μM) was also linear but was inhibited 30 min after initialization of the reaction. The concentration of quizalofop that inhibited 14C-acetate incorporation 50% (I50) was 0.54 μM. Chlorimuron, up to 155 μM, had no effect on 14C-acetate incorporation. A mixture of quizalofop (1.1 μM) and chlorimuron (4.8 μM) inhibited 14C-acetate incorporation similar to that of quizalofop alone at 1.1 μM. Quizalofop I50 for incorporation of 14C-pyruvate was 1.1 μM, and clorimuron at 4.8 μM decreased incorporation 15%. Excess unlabeled pyruvate (5 μM) had no effect on either 14C-acetate or 14C-pyruvate incorporation in the presence of both herbicides. It is believed that antagonism of quizalofop by clorimuron is not due to an excess pool of pyruvate resulting from inhibition of acetolactate synthase by chlorimuron.

The effects of pyruvate (Pyr), creatine pyruvate (Cr-Pyr) and creatine (Cr) on lipid and protein metabolism were compared in broiler chickens. A total of 400 1-day-old male birds (Aconred) were allocated to four groups, each of which included four replicates (25 birds per replicate). Treatments consisted of unsupplemented basal diet (Control), basal diet containing 2% Pyr, basal diet containing 3% Cr and basal diet containing 5% Cr-Pyr. Cr-Pyr and Pyr significantly decreased the hepatic triglyceride and serum total cholesterol concentration (P < 0.01). Cr-Pyr markedly increased the serum non-esterified fatty acid and high-density lipoprotein cholesterol concentrations (P < 0.05), whereas the expression of carnitine palmitoyl transferase I (P < 0.05) and peroxisome proliferators-activated receptor-α (P < 0.01) mRNA in the liver were both decidedly enhanced in the Cr-Pyr group. The relative leg muscle weight was higher in the Cr-Pyr group than in the control group, whereas the serum uric acid content and hepatic glutamic-oxaloacetic transaminase activity were lower in the Cr-Pyr and Cr groups (P < 0.05), respectively. Muscle insulin-like growth factor I (P < 0.05) expression was enhanced, and the myostatin (P < 0.01) mRNA level was reduced in both the Cr-Pyr and Cr groups. In addition, Cr-Pyr did not alter body weight or the feed conversion ratio. These results indicate that, compared with Pyr and Cr alone, Cr-Pyr has a bifunctional role in broiler chickens, in that it influences both lipid and protein metabolism.

In the rods of Bufo marinus and other species, bleaching of the rhodopsin in isolated cells leads to a loss of sensitivity and response amplitude and to a shortened response duration. These changes are permanent for cells bathed in Ringer&s solution. They are due to as yet unknown modulations in the transduction biochemistry. In this paper, we report that these changes can be partly or completely reversed by supplying biotin, pyruvate, and elevated glucose to the rod. The time course of this reversal and the substances which promote it imply that these are metabolically mediated effects. Based on the reported action of biotin and pyruvate on the one hand and on the changes of the response waveforms on the other hand, we believe that the phenomena we observe involve the later steps of the transduction cycle.

This study was conducted to examine the effect of energy substrates in a serum-free culture medium on in vitro development of porcine embryos. Presumptive zygotes derived from in vitro fertilization were cultured in glucose-free North Carolina State University (NCSU)-23 medium with glucose, pyruvate, fructose and lactate added to the culture medium singly or in various combinations. In experiment 1, a higher percentage of embryos cleaved (53–63% vs 10–13%) and developed to the blastocyst stage (18–27% vs 0) after the single addition of glucose (5.6 mM), pyruvate (0.5 mM) or lactate (10 mM) than with no energy substrate addition or the addition only of fructose (5.6 mM). In experiment 2, the addition of pyruvate and lactate resulted in higher blastocyst formation (25%) than other combinations (6–22%), while the addition of glucose and pyruvate significantly inhibited blastocyst formation. Increasing lactate concentration, as a single energy supplement, from 5 to 20 mM significantly improved blastocyst formation (7% vs 14–18%), while no benefit was achieved from increasing pyruvate concentration up to 2 mM (experiment 3). Glucose-free NCSU-23 medium supplemented with 0.5 mM pyruvate and 5 mM lactate significantly improved blastocyst formation (28% vs 17%) compared with NCSU-23 medium supplemented with 5.6 mM glucose (experiment 4). In conclusion, pyruvate and lactate are preferable energy substrates to support in vitro development of porcine embryos cultured in a serum-free NCSU-23 medium.

This study was carried out to examine how different combinations of pyruvate and glucose affect spontaneous meiotic maturation of cumulus-cell-enclosed mouse oocytes (CEO) to metaphase II (MII). Most experiments used an open system in which oocytes were cultured in 1 ml medium in plastic tubes. Initial experiments examined the dose response effects of pyruvate or glucose alone in the presence or absence of 2 mM glutamine. When medium lacked both pyruvate and glucose, more than 91% of the oocytes died in glutamine-free medium during 15 h of culture; viability was restored with the addition of glutamine, but only 11% of the CEO reached MII. In the absence of glutamine, 62–68% of oocytes completed maturation in 0.23–2.3 mM pyruvate, while 44–60% MII was observed in 0.55–27.8 mM glucose. The addition of glutamine to these cultures had a general suppressive effect on the completion of maturation. When glucose was added to pyruvate-containing cultures, the combination of 1 mM pyruvate/5.5 mM glucose was most effective in supporting maturation (about 90% MII), with little effect of glutamine. No further increase in maturation was observed when glucose was increased five-fold (to 27.8 mM). The positive effect of glucose was in part attributed to stimulation of glycolysis and increased production of pyruvate, since a reduced culture volume (8 μl), which allows the accumulation of secreted pyruvate, improved maturation in glucose-containing, but not pyruvate-containing, medium, and FSH, which stimulates glycolysis, increased progression to MII in glucose-containing, but not pyruvate-containing, medium. Yet these results also suggest that glucose has a beneficial effect on maturation apart from simple provision of pyruvate. The pyruvate effect was directly on the oocyte, because denuded oocytes responded more effectively than CEO to this energy substrate. The highest percentage of MII oocytes (96–97%) occurred in microdrop cultures containing glucose but lacking glutamine. These results indicate that glutamine supports oocyte viability but is not an adequate energy source for the completion of spontaneous meiotic maturation and may be detrimental. In addition, while pyruvate and glucose alone can each support meiotic progression of CEO to MII, optimal maturation requires the provision of both substrates to the culture medium when a large volume (1 ml) is used. It is concluded that careful attention to specific energy substrate supplementation and culture volume is important to optimise spontaneous meiotic maturation in vitro.