85 results about "Tetrahydrofolates" patented technology

The invention provides a non-naturally occurring microbial organism having an acetyl-CoA pathway and the capability of utilizing syngas or syngas and methanol. In one embodiment, the invention provides a non-naturally occurring microorganism, comprising one or more exogenous proteins conferring to the microorganism a pathway to convert CO, CO₂ and/or H₂ to acetyl-coenzyme A (acetyl-CoA), methyl tetrahydrofolate (methyl-THF) or other desired products, wherein the microorganism lacks the ability to convert CO or CO₂ and H₂ to acetyl-CoA or methyl-THF in the absence of the one or more exogenous proteins. For example, the microbial organism can contain at least one exogenous nucleic acid encoding an enzyme or protein in an acetyl-CoA pathway. The microbial organism is capable of utilizing synthesis gases comprising CO, CO₂ and/or H₂, alone or in combination with methanol, to produce acetyl-CoA. The invention additionally provides a method for producing acetyl-CoA, for example, by culturing an acetyl-CoA producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding an acetyl-CoA pathway enzyme or protein in a sufficient amount to produce acetyl-CoA, under conditions and for a sufficient period of time to produce acetyl-CoA.

Described herein is a method for reducing levels of the harmful metabolic waste product of S-adenosylmethionine (SAMe), homocysteine, and provide vitamin and other nutritional co-factors that reduce the production of homocysteine and either re-methylate homocysteine back to S-adenosylmethionine, or facilitate its conversion downstream to cystathione. The method of the invention may be achieved by administering 5-methyl tetrahydrofolate, methylcobalamin, and one or more compounds selected from the group consisting of betaine, pyridoxal-5-phosphate, N-acetyl-cysteine, and other cofactors.

The present invention provides novel uses and compositions for 5,10-methylene tetrahydrofolate (“5,10-CH₂-THFA”) in the treatment of cancer. The present invention is based on the surprising result that 5,10-CH₂-THFA, while increasing the efficacy of 5-fluoruracil (5-FU) in reducing the rate of tumor growth and increasing survivorship, also reduces the toxicity to the patient of 5-FU. The present invention provides methods and compositions for treating cancer patients that include 5-FU, 5,10-CH₂-THFA, and one or more additional anticancer drugs. Such methods and compositions can provide increased efficacy and reduced toxicity when compared with current treatment modalities.

The invention discloses a kit for quantitative guidance of folic acid supplement dosage before and during pregnancy and application thereof. The kit comprises a specific primer pair, a specific fluorescent probe pair, a convention assembly and the like, wherein the specific primer pair and a specific fluorescent probe pair can simultaneously detect three single nucleotide polymorphism genotypes on a 5, 10-methylene tetrahydrofolate reductase (MTHFR) gene and a methionine synthase reductase (MTRR) gene, and the convention assembly is used to for fluorescent quantitative PCR detection. The kit of the invention can be used for the quantitative guidance of the folic acid supplement dosage before and during the pregnancy.

The present invention relates to a pharmaceutical composition which may comprise progestogens, preferably drospirenone, estrogens, preferably ethinylestradiol and 5-methyl-(6S)-tetrahydrofolate, which may be employed as oral contraceptive and moreover prevents disorders caused by folate deficiency in the consumers, in particular cardiovascular disorders and, after conception of the embryo, congenital malformations caused by folate deficiency such as, for example, neural tube defects, ventricular valve defects, urogenital defects, and cleft lip, jaw and palate, without masking the symptoms of vitamin B₁₂ deficiency, and at the same time even in the case of homozygous or heterozygous polymorphism of methylenetetrahydrofolate reductase facilitates unimpaired utilizability of the folate component 5-methyl-(6S)-tetrahydrofolate by the body and thus its biological activity for preventing the abovementioned congenital malformations caused by folate deficiency. In addition, a prolonged protective effect is maintained after discontinuation of the contraceptive.

The present invention describes drug delivery compositions in the form of thin water-soluble films (wafers), which contain particles that comprise at least one active ingredient—which is not an estrogen and/or a progestin and/or an alkaline earth metal salt of 5-methyl-(6S)-tetrahydrofolate—and at least one protective agent. The protective agent provides effective taste-masking of the active ingredient due to limited release of the active ingredient in the mouth. The active ingredient is hence not absorbed via the buccal route, but rather via the enteral (per-oral) route. The particles contained in the wafer provided by the present invention have a particle size of below 40 μm thereby resulting in an acceptable sensation in the mouth while dissolving. Such wafers are especially suitable for pediatric use.

The invention discloses a high-yield 5-methyl tetrahydrofolate recombinant bacillus subtilis and an application thereof, and belongs to the field of genetic engineering. The high-yield 5-methyl tetrahydrofolate recombinant bacillus subtilis uses the bacillus subtilis 168 as the expression host; by over-expresseing 5,10-methylene tetrahydrofolate reductase coding gene metF, dihydrofolate reductasecoding gene dfrA, dihydrofolate synthase coding gene folC, knocking-out double function homocysteine S-methyltransferase/5,10-methylene tetrahydrofolate reductase coding gene yitJ, formyltetrahydrofolate deformylase coding gene purU, the high-yield 5-methyl tetrahydrofolate recombinant bacillus subtilis obtains accumulation of 5-methyl tetrahydrofolate bacillus subtilis gene engineering bacteria;the yield of 5-methyl tetrahydrofolate reached 952.05 ug/ L, and the conversion cell dry weight is 342.40ug/ g DCW.

Preeclampsia and intrauterine growth restriction in a pregnant female mammal are prevented or decreased in severity by administering thereto a combination of a vitamin compound containing B1, Folic Acid (or active form 5-Methyl-Tetrahydrofolate i.e.; Metafolin®), B 6 (Pyridoxine or active form Pyridoxine 5-Phosphate P5P), B 12, Ascorbic Acid, Selenium, Zinc, Co-enzyme Q 10 and N-Acytyl Cysteine, Lycopene, optionally in further combination with Melatonin and/or Vitamin E or/and ASA 81 mg both of later, are cyclooxygenase inhibitors, a PGI.sub.2-mimetic, a thromboxane (TXA.sub.2) inhibitor, a compound possessing TXA.sub.2-agonistic and TXA.sub.2-inhibiting properties, a compound possessing TXA.sub.2-antagonistic and PGI.sub.2-mimetic activities, and a TXA.sub.2 antagonist.

The invention relates to an application of molecular beacon probes and a melting curve technology in detection of gene polymorphism, in particular to a method for detecting MTHFR (methylene tetrahydrofolate reductase) and MTRR (methionine synthase reductase) gene polymorphism simultaneously, and further relates to amplification primers of MTHFR and MTRR genes, the molecular beacon probes and a kitcontaining the amplification primers and the molecular beacon probes. The method is simple and fast to operate, good in specificity, high in sensitivity, accuracy and flux and low in detection cost and has broad application prospect. According to the method, PCR amplification and detection are performed synchronously, the overall detection process does not require uncovering operation, so that detection period is shortened and detection efficiency is improved while detection cost is saved, and besides, risk of false positive caused by PCR product pollution is reduced.

The invention discloses a kit for investigating polymorphism of an MTHFR (Methylene Tetrahydrofolate Reductase) and/or MTRR (Methylenetetrahydrofolate Reductase) gene on the basis of a Taqman-MGB probe. The kit comprises a primer group and a probe, wherein the primer group is at least one selected from the following three groups: primers of SEQ ID NO.1 and SEQ ID NO.4 for an MTHFR gene at a C677Tsite, primers of SEQ ID NO.7 and SEQ ID NO.10 for an MTRR gene at an A1298C site, primers of SEQ ID NO.13 and SEQ ID NO.14 for an MTRR gene at an A66G site, primers of SEQ ID NO.22 and SEQ ID NO.19, primers of SEQ ID NO.29 and SEQ ID NO.15, and primers of SEQ ID NO.33 and SEQ ID NO.30. The kit disclosed by the invention has the advantages that three mutation sites can be detected simultaneously, high sensitivity is achieved, plasma as low as 10copies can be accurately detected, and oral cavity swabs which are too long in preservation time or relatively low in concentration can be still accurately detected.

The invention provides a gene noninvasive detection kit for preventing neural tube defects of newborns. The kit comprises specific primers, DNA sequencing primers, a PCR (Polymerase Chain Reaction) reaction assembly, a PCR product purification assembly, a DNA sequencing reaction assembly and the like, wherein the specific primers are used for detecting No. rs1801133 SNP locus (MTHFR C677T) and No. rs1801131 SNP locus (MTHFR A1298C) on 5, 10-methylene tetrahydrofolate reductase (MTHFR) and No. rs1801394 SNP locus (MTRR A66G) on 5, 10-methylene methyl transferase reductase (MTRR). The kit evaluates the risk level of pregnant women giving birth to infants with neural tube defects by detecting the genotype of a single nucleotide polymorphism locus closely related to the main genetic factor causing newborn defects, namely female folic acid metabolism ability, and the pregnant women are individually instructed to supplement folic acid according to the gene detection result of each client. The method provided by the invention accords with the situation of our country, oral mucosa cell sampling is adopted as a sampling method which is painless and noninvasive, and cross infection is avoided. The sequencing detection results are accurate and reliable, expensive imported special instruments are not needed, and the method is easy to popularize and spread.

The invention discloses a preparation method of L-5-calcium methyltetrahydrofolate. The preparation method comprises the following steps of (1) adopting folic acid as a raw material, and reducing to preparing an intermediate 6-(R,S)-tetrahydrofolic acid; (2) carrying out chiral resolution on the 6-(R,S)-tetrahydrofolic acid, and obtaining an intermediate 6-S-tetrahydrofolate; (3) carrying out methylation on the 6-S-tetrahydrofolate to obtain an intermediate L-5-methyltetrahydrofolate; (4) reacting the L-5-methyltetrahydrofolate and salt containing calcium to obtain the L-5-calcium methyltetrahydrofolate. According to the method provided by the invention, the problem that hydrogen, wastewater and waste residues are easily generated by using a large number of sodium borohydride is avoided.

The invention relates to a folic acid metabolic capability and genotyping detection technology and in particular relates to a kit and a detection method of a folic acid metabolic capability and genotyping as well as application of the kit. The kit comprises specific primers aiming at 677C>T mutation and 1298A>C mutation of an MTHFR (Methylene Tetrahydrofolate Reductase) gene, and 66A>G mutation ofan MTRR (Methionine Synthase Reductase) gene, and a specific mutation detection probe, RT-PCR (Reverse Transcription-Polymerase Chain Reaction) buffer, an RT-PCR reaction Taq enzyme, sterile purifiedwater, a negative quality control product, a wild type quality control product, a mutant quality control product and a packaging box for separating and packaging a reagent bottle or pipe. The detection method of the folic acid metabolic capability and the genotyping, provided by the invention, has the advantages of strong specificity, high sensitivity, small pollution, simplicity and rapidness inoperation, high safety performance and the like; a detection result has relatively good accuracy and repeatability; the detection method is especially suitable for directly taking whole blood as a detection sample and related gene mutation detection is directly carried out; a genome DNA (Deoxyribonucleic Acid) template does not need to be extracted. The folic acid metabolic capability can be accurately judged and individualized folic acid oral administration and supplementing dosages are provided; the kit and the detection method have important value.

The invention discloses a kit for detecting an individual folate metabolism disorder. The kit comprises a specific primer pair and SYBRGreenI fluorochrome for simultaneously detecting rs1801133# SNP site on a 5,10-methylene tetrahydrofolate reductase gene, rs1801131# SNP on a 5,10-methylene tetrahydrofolate reductase gene and rs1801394# SNP site on a methionine synthase reductase gene, a conventional component for real-time quantitative polymerse chain reaction (PCR) detection. The kit disclosed by the invention evaluates the metabolic capability of individual folate by simultaneously detecting single nucleotide polymorphism site genotype of the 5,10-methylene tetrahydrofolate reductase gene and the methionine synthase reductase gene of folate metabolism.

The invention discloses a stable pharmaceutical composition, which contains (6S)-5-methyl-calcium tetrahydrofolate crystal and/or pharmaceutically acceptable reducing substances, and pharmaceutically acceptable auxiliary materials. Specifically, the reducing substances can protect (6S)-5-methyl-tetrahydrofolic acid and its salt from being oxidized, and can be selected from vitamin C and its salt, isovitamin C and its salt, mercaptoethanol, cysteine, mercaptoethyl sulfonic acid, dithiothreitol, reduced glutathione, and lipoic acid. The stable pharmaceutical composition provided by the invention has good stability during processing and storage, the drug risk caused by degradation can be avoided, and at the same time fast and reliable release of (6S)-5-methyl-tetrahydrofolic acid in the composition can be ensured.

There is provided is a nucleotide primer set for LAMP amplification used for detecting genotypes of single-nucleotide polymorphisms C677T and A1298C of an MTHFR gene. There is also provided a nucleotide probe for detecting an amplification product amplified by the primer set according to the present invention. There is also provided a method of detecting the genotypes of the single-nucleotide polymorphisms C677T and A1298C in the MTHFR gene, by using the primer set according to the present invention.

The present invention relates to a pharmaceutical composition which comprises progestogens, preferably drospirenone, estrogens, preferably ethinylestradiol and 5-methyl-(6S)-tetrahydrofolate, can be employed as oral contraceptive and moreover prevents disorders caused by folate deficiency in the consumers, in particular cardiovascular disorders and, after conception of the embryo, congenital malformations caused by folate deficiency such as, for example, neural tube defects, ventricular valve defects, urogenital defects, and cleft lip, jaw and palate, without masking the symptoms of vitamin B₁₂ deficiency, and at the same time even in the case of homozygous or heterozygous polymorphism of methylenetetrahydrofolate reductase facilitates unimpaired utilizability of the folate component 5-methyl-(6S)-tetrahydrofolate by the body and thus its biological activity for preventing the abovementioned congenital malformations caused by folate deficiency. In addition, a prolonged protective effect is maintained after discontinuation of the contraceptive.

The invention discloses a (6S)-5-methyl tetrahydrofolate crystal form and a preparation method thereof. The crystal form is a type C calcium (6S)-5-methyl tetrahydrofolate crystal form; the X-ray diffraction spectrum has diffraction peaks at angles 2 theta, namely 6.3+/-0.2 and 19.2+/-0.2. The type C calcium (6S)-5-methyl tetrahydrofolate crystal form has the advantages of excellent physicochemical properties, good stability, high purity, good reproducibility, better suitability for industrial large-scale preparation, and the like.

The invention relates to a detection method and a kit for coronary heart disease susceptibility loci. The method comprises the following steps: (1) extracting a genome deoxyribonucleic acid (DNA) of a sample, and amplifying a region containing rs1801133 loci in a methylene tetrahydrofolate reductase (MTHFR) gene exon of the sample, so as to obtain an amplification product; (2) detecting the genetype of single nucleotide polymorphism (SNP) loci rs1801133 in the amplification product by using a high resolution melting (HRM) analysis technology. The genetype analysis is carried out on the region containing the rs1801133 loci in the amplified MTHFR gene by adopting the HRM analysis technology. The method is simple and quick to operate, low in use cost and accurate in result, and can achieve batch inspection; whether the tested people carry with coronary heart disease susceptibility genes can be clearly judged by using the kit, the coronary heart disease susceptibility people can be screened out from people, the poor living habit is improved, and the target of prevention is achieved.

The present invention relates to a pregestational gene alarm diagnosis kit and its detection method, including DNA extraction liquor for extracting DNA template of testee, PCR mixed liquor for amplifying specific fragment on the genomic DNA of testee, DNA polymerase, PCR product enzyme excision mixed liquor, restriction endonuclease, positive control template of homozygous mutation and coupling pipe for making PCR reaction. The described alarm gene includes N5, N10-methylene tetrahydrofolate reductase and methionine synthetase reductase gene, their mulant sites are respectively 667-cytosine-thymine and 66 adenine-guanine, and their primer sequences respectively are 5'-TGA AGG AGA AGG TGT CTG CGG GA-3', 5'-AGG ACG GTG CGG TGA GAG TG-3', 5'-GCA AAG GCC ATC GCA GAA GAC AT-3' and 5'-GTG AAG ATC TGC AGA AAA TCC ATG TA-3'. It can be used for pregestational diagnosis of that filial generation produces neural tube deform or not.