Exploration of Indole Alkaloids from Marine Fungus Pseudallescheria boydii F44-1 Using an Amino Acid-Directed Strategy

The composition of the culture medium has great influence on the metabolite production of the marine fungus Pseudallescheria boydii F44-1. By adding amino acids to GPY culture medium, two new bisindole alkaloids, pseudboindoles A and B (1 and 2), together with 11 known indole alkaloids were isolated from the culture broth. Their structures were elucidated by comprehensive analysis of the NMR, MS, IR, and UV spectra. The 3,3′-cyclohexylidenebis(1H-indole) (3) showed cytotoxic activity against various cancer cell lines.

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P–283 Combination between amino acids profile of the spent culture media and morphokinetics parameters of human embryos to determine its viability

Human Reproduction ◽

10.1093/humrep/deab130.282 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

N Adel ◽

M Kadah ◽

S Abdulghafar ◽

M Elmahdy ◽

D Ghareeb ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Embryo Development ◽

Culture Medium ◽

Essential Amino Acids ◽

Human Embryos ◽

Data Set ◽

Group A ◽

Amino Acids Profile ◽

Group D

Abstract Study question How to determine human embryo viability noninvasively before embryo transfer? Summary answer We propose that the combination of the amino acid profile of an individual embryo with its morphokinetics will provide noninvasive tool to determine its viability. What is known already It was already known that human embryos at early cleavage require non-essential amino acids, while at the 8-cell to blastocyst stages, a mixture of non-essential and essential amino acids. Amino acids have important roles during embryo development. Acting as biosynthetic precursors,buffers of intracellular pH in the embryo, antioxidants, energy sources and regulators of metabolic function and signaling pathways. Many studies have used time-lapse to analyze human embryonic development including the process of fertilization and assessment of early events and introducednoninvasive prognostic markers which predict embryo development and correlate it to IVF treatment outcomes. Study design, size, duration This study was a prospective cohort study approved by the Clinical Trial Ethical Committee of Faculty of Medicine, Alexandria University according to ethical standards of scientific research (Serial number: 0303721).Thirty females aged 30.13 ± 4.83 years undergoing ICSI cycle in the Madina Fertility Center, during the period of March 2018 to November 2019.202 MII oocytes were incubated individually in embryoscope. Participants/materials, setting, methods Embryos (n = 161) were divided on Day 5 into two groups –developed embryos “Group D” (embryos that developed to blastocyst) and arrested embryos “Group A” (embryos remain at cleavage stage and fail to develop to blastocys).Developed embryos (Group D) included 99 embryos, and Arrested embryos (Group A) included 62 embryos. For each group, morphokinetic developmental points using embryoscope and the different amino acids concentrations in spent culture medium were analyzed using LC- mass spectro etry. Main results and the role of chance On one hand, the first appearance of pronuclei (TPNa), t2, t4 and CC2 in group D occurred significantly earlier than those of Group A.Analysis of 19 essential and non-essential amino acids in spent culture medium of each embryo in the two studied groups D and A showed a significantly higher concentration of two essential amino acids L-Valine (145.73 ± 150.96) and L-Phenylalanine (61.59 ± 55.78) in Group D than their concentration in Group A ( 104.58 ± 33.58, 44.24 ± 14.61, respectively , p ≤ 0.05).and significantly lower concentration of three non-essential amino acids L-Tyrosine (62.56 ± 41.03) , L-Cysteine (19.48 ± 11.90), and L-Alanine (136.0 ± 389.83) observed in Group D when compared to Group A (69.57 ± 20.78, 22.37 ± 8.59,145.33 ± 165.22, respectively, Limitations, reasons for caution It is important to note, that results were developed on a data set from one clinic with different stimulation protocols, a multicenter data and a correlation with the stimulation protocol used should be involved in future studies, in addition a larger sample size to avoid high standard deviation is recommended Wider implications of the findings: We can conclude that amino acid turnover is independent of the traditional morphological assessment of embryos and it may reflect its viability. The prospective combined use of amino acids profile of individual embryo and its morphokinetic parameters may contribute to introduce a new noninvasivs tool that may improve implantation rate Trial registration number 0303721

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Amino acid metabolism of bovine blastocysts derived from parthenogenetically activated or in vitro fertilized oocytes

Reproduction Fertility and Development ◽

10.1071/r98052 ◽

1998 ◽

Vol 10 (3) ◽

pp. 279 ◽

Cited By ~ 10

Author(s):

Y. G. Jung ◽

T. Sakata ◽

E. S. Lee ◽

Y. Fukui

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Metabolism ◽

Culture Medium ◽

Acid Metabolism ◽

Essential Amino Acids ◽

Fluid Medium ◽

Vitro Fertilization ◽

Oviduct Fluid

The uptake and synthesis of 19 amino acids by fresh or frozen–thawed bovine blastocysts produced by parthenogenesis (PT) or in vitro fertilization (IVF) were compared in the present study. Fresh blastocysts, 180 h after IVF or PT activation, and frozen–thawed blastocysts, 168 h old and cultured for 12 h post-thawing, were cultured in synthetic oviduct fluid medium (SOFM) containing polyvinyl alcohol (PVA) with both essential and non-essential amino acids (EAA and NEAA, respectively) (Medium 1: M1) or SOFM containing PVA with only EAA (Medium 2: M2). In Experiment 1, when fresh or frozen–thawed PT blastocysts were cultured in M1, the uptake of glutamate (in fresh only), aspartate and arginine, and the synthesis of glutamine and alanine were significantly enhanced. In the culture with M2, serine, asparagine, glutamate, glutamine, glycine, arginine and alanine were significantly taken up. It was found that the glutamine concentrations was significantly higher (P < 0.001) in the culture medium drops containing embryos than in the drops without embryos. In Experiment 2, when PT blastocysts were cultured in M1, the uptake of aspartate and synthesis of alanine were greater (P < 0.01) than those by IVF blastocysts. When M2 was used, a significant (P < 0.01) production of serine, asparagine, glutamate, glutamine and alanine, and the uptake of arginine by PT blastocysts were observed. In Experiment 3, when IVF blastocysts were cultured in M1, fresh blastocysts depleted more aspartate and glutamate, and produced more glutamine and alanine than frozen–thawed blastocysts. When cultured in M2, frozen–thawed blastocysts depleted more threonine (P < 0.01) than fresh blastocysts. These results indicate that the uptake and synthesis of amino acids were different in fresh or frozen–thawed bovine blastocysts derived from PT or IVF. These differences in amino acid metabolism may be related to the viability of the blastocysts.

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Effects of an Electric Field of Sinusoidal Waves on the Amino Acid Biosynthesis by Azotobacter

Zeitschrift für Naturforschung C ◽

10.1515/znc-1980-3-413 ◽

1980 ◽

Vol 35 (3-4) ◽

pp. 258-261

Author(s):

A. Martin Gonzalez ◽

M. T. Izquierdo

Keyword(s):

Amino Acids ◽

Nitrogen Fixation ◽

Amino Acid ◽

Electric Field ◽

Electric Fields ◽

Culture Medium ◽

Azotobacter Vinelandii ◽

Amino Acid Biosynthesis ◽

Medium Composition ◽

Acid Biosynthesis

Abstract Electric Field Electric fields of sinusoidal waves have been applied in cultures of Azotobacter vinelandii, with potentials between 0 V and 10 V, intensities from 0 mA to 16 mA and frequencies between 5 Hz and 200 KHz. The influence of the electric field of sinusoidal waves on the nitrogen fixation on the post culture medium composition has a maximum at 5 V, 8 mA and 20 Hz. The rate of synthesis of specific amino acids by Azotobacter depends on the frequency and potential of the electric field applied. The concentration of each amino acid present in the post-culture medium is increased according to the electric field employed and the amino acid biosynthesis in culture medium is activated during the first days of incubation.

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Effects of amino acids on development in vitro of cleavage-stage bovine embryos into blastocysts

Reproduction Fertility and Development ◽

10.1071/rd9960835 ◽

1996 ◽

Vol 8 (5) ◽

pp. 835 ◽

Cited By ~ 28

Author(s):

T Pinyopummintr ◽

BD Bavister

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Embryo Development ◽

Culture Medium ◽

Essential Amino Acids ◽

Bovine Embryo ◽

Bovine Embryos ◽

Development In Vitro ◽

Defined Culture

Effects of amino acids on early bovine embryo development in vitro were examined using a chemically-defined, protein-free culture medium. Bovine embryos produced in vitro were cultured from 18 h to 72 h post insemination in a simple medium containing lactate as the only energy source except for the amino acid treatments. Subsequently, embryos were transferred to TCM-199 supplemented with serum for blastocyst development to substantiate their developmental competence. Treatments were: (1) non-essential amino acids from TCM-199 (NEA); (2) essential amino acids from TCM-199 (EA); (3) NEA+EA; (4) Eagle's minimum essential medium amino acids (MEM AA); (5) 11 amino acids present in HECM-6 (11 AA); and (6) 0.2 mM glutamine (GLN). A higher proportion of embryos (percentage of inseminated ova) cleaved to the > or = 8-cell stage by 72 h post insemination in NEA (56.7%), EA (41.2%), 11 AA (40.3%) and GLN (51.1%) than in either NEA+EA (30.0%) or MEM AA (33.1%). However, after transfer to complex medium, embryos that had developed in EA, as well as those in MEM AA or NEA+EA, produced significantly fewer blastocysts (37.1%, 34.4% and 25.6% respectively) than those in NEA (56.7%), GLN (48.9%) or 11 AA (37.7%). The ability of blastocysts to hatch from their zonae pellucidae was also affected by amino acid treatment during cleavage stages. The present study indicated that the addition of NEA or GLN or 11 AA to a chemically-defined culture medium during the cleavage phase of bovine embryo development increases their subsequent ability to reach the blastocyst stage. These data have implications for understanding the nutritional needs of bovine embryos produced in vitro and for optimizing the composition of culture media to support their development.

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Amino Acid Metabolism in Human Embryos

Physiological Research ◽

10.33549/physiolres.933240 ◽

2016 ◽

pp. 823-832 ◽

Cited By ~ 3

Author(s):

P. DRÁBKOVÁ ◽

L. ANDRLOVÁ ◽

R. HAMPL ◽

R. KANĎÁR

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Metabolism ◽

Culture Medium ◽

Acid Metabolism ◽

Culture Media ◽

Time Lapse ◽

Human Embryos ◽

Strongly Correlated ◽

Cell Stage

The aim of this study was to find some relationship between amino acid metabolism and the embryo morphokinetic parameters studied via time-lapse analysis. Study included 48 human embryo samples and their culture media. Two groups of embryos were identified: embryos reached the 8-cell stage on day 3 (n=34) and embryos failed to develop at any point during the incubation (n=14). Amino acids levels were measured on day 3 of embryo development; using time-lapse analysis, the precise timing of embryo cleavage, synchrony of division, grade of fragmentation etc. were established. No statistically significant differences between dividing and arresting embryos were observed in terms of amino acids production/consumption and turnover. Amino acids which were part of the culture medium did not exhibit any statistically significant correlation with kinetic parameters with the exception of the grade of fragmentation on day 3; there were negative correlation with glutamate, and positive with glutamine, glycine and taurine. In some dividing and in some arresting embryos appeared new amino acids which strongly correlated with each other, with methionine, but not with any other amino acid that is a regular part of the culture medium.

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Cloning of rat asparagine synthetase and specificity of the amino acid-dependent control of its mRNA content

Biochemical Journal ◽

10.1042/bj3040745 ◽

1994 ◽

Vol 304 (3) ◽

pp. 745-750 ◽

Cited By ~ 46

Author(s):

R G Hutson ◽

M S Kilberg

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Culture Medium ◽

Fold Increase ◽

Trna Synthetase ◽

Asparagine Synthetase ◽

Single Amino Acid ◽

General Control ◽

Nutrient Regulation ◽

Mrna Content

A full-length cDNA clone for rat asparagine synthetase (AS) was isolated from a cDNA library enriched for amino acid-regulated sequences. The AS cDNA was used to investigate the amino acid-dependent repression of AS mRNA content in rat Fao hepatoma cells. In response to complete amino acid starvation, there was an approximately 10-fold increase in the level of AS mRNA. Three species of mRNA, of approx. sizes 2.0, 2.5 and 4.0 kb, were detected and each was simultaneously regulated to the same degree. The expression of AS mRNA increased by 6 h after removal of amino acids, reached a plateau after 9 h, and was blocked by either actinomycin D or cycloheximide. Partial repression of the AS mRNA content was maintained by the presence of a single amino acid in the culture medium, but the degree of effectiveness for each one varied widely. Glutamine showed the greatest ability to repress the AS mRNA content, even at an extracellular concentration 10 times below its plasma level. Other effective repressors included the amino acids asparagine, histidine and leucine, as well as ammonia. Depletion of selected single amino acids from an otherwise complete culture medium also caused up-regulation. In particular, removal of histidine, threonine or tryptophan from the medium, or the addition of histidinol to inhibit histidinyl-tRNA synthetase, resulted in a significant increase in AS mRNA content. The data indicate that nutrient regulation of AS mRNA occurs by a general control mechanism that is responsive to a spectrum of amino acids.

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Alkaloids From the Marine Fungus Lecanicillium fusisporum Using an Amino Acid-Directed Strategy

Natural Product Communications ◽

10.1177/1934578x211046072 ◽

2021 ◽

Vol 16 (10) ◽

pp. 1934578X2110460

Author(s):

Qin Huang ◽

Hou-Jin Li ◽

Chang-Bai Huang ◽

Zi-Han Wang ◽

Wen-Jian Lan ◽

...

Keyword(s):

Amino Acid ◽

Spectroscopic Data ◽

Marine Fungi ◽

Optical Rotation ◽

Indole Alkaloid ◽

2D Nmr ◽

Marine Fungus ◽

Culture Broth ◽

Synthetic Approach ◽

C Nmr

An amino acid-directed strategy has been developed to explore the potential of marine fungi to produce alkaloids. The marine fungus Lecanicillium fusisporum was cultured in glucose-peptone-yeast (GPY) medium to which were added L-tryptophan, L-phenylalanine, L-threonine, D, L-methionine, L-serine, L-lysine and L-valine. A new indole alkaloid, lecasporinoid (1), along with five known alkaloids (2−6) were discovered from the culture broth. The planar structure of lecasporinoid (1) was determined by HR-ESIMS, and 1D and 2D NMR spectroscopic data. The absolute configuration was established by optical rotation and 13C NMR calculations combining with a chemical synthetic approach.

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Regulation of enzyme turnover during tissue differentiation. Interactions of insulin, prolactin and cortisol in controlling the turnover of fatty acid synthetase in rabbit mammary gland in organ culture

Biochemical Journal ◽

10.1042/bj1540359 ◽

1976 ◽

Vol 154 (2) ◽

pp. 359-370 ◽

Cited By ~ 43

Author(s):

Brian K. Speake ◽

Raymond Dils ◽

R. John Mayer

Keyword(s):

Amino Acids ◽

Protein Synthesis ◽

Fatty Acid ◽

Amino Acid ◽

Culture Medium ◽

Fatty Acid Synthetase ◽

Amino Acid Pool ◽

Acid Pool ◽

Intracellular Amino Acid ◽

Enzyme Degradation

1. Explants of mammary gland from mid-pregnant rabbits were cultured in Medium 199 containing combinations of insulin, prolactin and cortisol. With hormone combinations which included prolactin, a sustained increase in the apparent rate of synthesis and in the amount of fatty acid synthetase was measurable immunologically. Maximum increase was produced with insulin, prolactin and cortisol present together. 2. With prolactin present alone, synthetase activity in the explants decreased to undetectable values after 1 day in culture, whereas the incorporation of l-[U-14C]leucine into immunodetectable material increased. Prolactin may therefore direct the synthesis of immunologically cross-reactive precursors of fatty acid synthetase which are enzymically inactive. 3. Culture with dibutyryl cyclic AMP plus theophylline in the presence of insulin, prolactin and cortisol delayed the increase in the rate of synthesis and accumulation of the synthetase. These compounds may also prevent the apparent decrease in the rate of degradation of the synthetase which occurs on day 2 of culture. 4. A large decrease in the apparent rate of degradation of the synthetase on day 2 of culture occurs during culture with hormone combinations which include prolactin. The protein obtained by centrifugation of explant homogenates for 6min at 14000gav. is degraded continuously throughout the culture period. 5. This decrease in the apparent rate of degradation of the synthetase was measured by radio-immunological precipitation. It is probably part of a regulated programme of enzyme degradation and not a reflexion of the reutilization of radioactive amino acids for the following reasons. (a) The calculated increase in the amount of the synthetase in explants on day 2 of culture with insulin, prolactin and cortisol was approximately equal to the measured increase of the enzyme complex which accumulates in the explants. This suggests little or no enzyme degradation has occurred. (b) Explants were cultured for 24h with insulin, prolactin and cortisol. They were then incubated with l-[U-14C]leucine, washed and incubated again for up to 4½h. l-[U-14C]Leucine rapidly equilibrated with the intracellular amino acid pool. Within 10min of incubation after washing explants to remove endogenous l-[U-14C]leucine the previously linear incorporation of l-[U-14C]-leucine into total explant protein ceased. This suggests that protein is synthesized from an amino acid pool which rapidly equilibrates with amino acids in the culture medium. (c) Explants were cultured for 24h as described in (b) but after washing they were cultured with insulin, prolactin and cortisol for 24h. Approx. 90% of the radioactivity lost from the ‘free’ intracellular amino acid pool and from amino acids derived from the degradation of explant protein in this period was detected in the culture medium. This suggests that the ‘free’ intracellular amino acids and amino acids derived from protein degradation can equilibrate with amino acids in the medium. A residual ‘free’ radioactive amino acid pool was present in the tissue. (d) Casein represents approx. 20% of the protein synthesized after 1 day in culture with insulin, prolactin and cortisol. Histological evidence suggests that on day 2 of culture, casein is unlikely to be degraded in the tissue. No increase in the radioactivity incorporated into casein can be measured in the 23h after incubation of explants with l-[U-14C]leucine as described in (b). This suggests that the incorporation of radioactivity into proteins during culture after incubation with l-[U-14C]leucine is minimal. (e) Inhibition of protein synthesis in explants by cycloheximide after incubation with l-[U-14C]leucine does not reveal a latent continuous degradation of fatty acid synthetase on day 2 of culture which might have been masked by the high rates of protein synthesis and therefore the accumulation of the enzyme. 6. The conclusion is discussed that there is a real decrease (or even cessation) in the rate of degradation of fatty acid synthetase during the period when the enzyme accumulates in explants cultured with hormone combinations which contain prolactin.

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Photo-dissociation of self-assembled (anthracene-2-carbonyl)amino acid hydrogels

Chemical Communications ◽

10.1039/d0cc05292b ◽

2020 ◽

Vol 56 (89) ◽

pp. 13792-13795

Author(s):

Phillip R. A. Chivers ◽

Rebecca S. Dookie ◽

Julie E. Gough ◽

Simon J. Webb

Keyword(s):

Amino Acids ◽

Cell Culture ◽

Amino Acid ◽

Culture Medium ◽

Uv Light ◽

Cell Culture Medium ◽

Subsequent Exposure ◽

Self Assembled

Anthracene-amino acids self-assemble into supramolecular hydrogels upon the addition of salts in water or cell culture medium; subsequent exposure to 365 nm UV light disassembles the hydrogel by photodimerising the gelators.

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Development of Entomophaga aulicae protoplasts in synthetic eastern hemlock looper hemolymph

Canadian Journal of Microbiology ◽

10.1139/m89-045 ◽

1989 ◽

Vol 35 (2) ◽

pp. 304-308 ◽

Cited By ~ 2

Author(s):

Gary B. Dunphy ◽

Richard A. Nolan

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Culture Medium ◽

Eastern Hemlock ◽

Optimum Temperature ◽

Medium Ph ◽

Fungal Development ◽

Hyphal Bodies ◽

Hemlock Looper

The protoplast stages of Entomophaga aulicae grew in a medium based on the amino acid and amine composition of the hemolymph of the eastern hemlock looper, Lambdina fiscellaria fiscellaria. The optimum temperature and pH ranges for spindle-shaped protoplast growth were 15 to 25 °C and 5.5 to 6.5, respectively. Spindle-shaped protoplast development progressed asynchronously through the formation of mesoprotoplasts, elliptical mesoprotoplasts, and rod-shaped and spherical hyphal bodies. The production of these stages was not influenced by either medium pH or osmolality. Conidia were formed on conidiophores arising from spherical hyphal bodies near the surface of the culture medium. Changes in amino acid and amine levels during growth were correlated with the stage of fungal development. The major amino acids utilized during development included L-glutamine, L-asparagine, L-proline, L-leucine, L-isoleucine, L-tyrosine, and the amine O-phosphoethanolamine.Key words: Entomophaga aulicae, Lambdina fiscellaria fiscellaria, protoplasts, hemolymph, ninhydrin-positive compounds.

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