Abstract: A method for making a surfactant-based monolithic column is provided. The method comprises providing a mixture comprising at least one surfactant monomer, at least one crosslinker, at least one initiator, and at least one porogen and polymerizing the mixture to form the surfactant-based monolithic column. The present disclosure also provides a surfactant-based monolithic column, a method for separating molecules, and a process for preparing a surfactant monomer.

Abstract: The present invention provides antibiofilm composition comprising two or more agents selected from the group consisting of DispersinB™, 5-Fluorouracil, Deoxyribonuclease I and Proteinase K for preventing growth and proliferation of biofilm-embedded microorganisms in wound care, oral care, and disease-related infections and methods of treatment in mammals. The invention further provides methods for preparing medical devices, and wound care devices using an antibiofilm composition comprising two or more antimicrobial agents selected from the group consisting of DispersinB™, 5-Fluorouracil, Deoxyribonuclease I and Proteinase K.

Abstract: The present invention concerns in general PCR reaction mixtures comprising a mixture of hot-start primers and non-hot-start primers for a given target sequence, including multiplex PCR reaction mixtures; methods utilizing same for detection of one or more target polynucleotide sequences; and kits comprising same.

Abstract: A nicking endonuclease is described which has an amino acid sequence with at least 70% identity to SEQ ID NO:6 and comprising a mutation at least one of an arginine or gutamic acid corresponding to position 507 and position 546 respectively in SEQ ID NO:6.

Abstract: A method for detecting a mutation related to the gene encoding OAS1. This and other disclosed mutations correlate with resistance of humans to viral infection including hepatitis C. Also provided is a therapeutic agent consisting of a protein or polypeptide encoded by the mutated gene, or a polynucleotide encoding the protein or polypeptide. Inhibitors of human OAS1, including antisense oligonucleotides, methods, and compositions specific for human OAS1, are also provided.

Abstract: Method of preparing 1-CreI meganuclease variants having a modified cleavage specificity, variants obtainable by said method and their applications either for cleaving new DNA target or for genetic engineering and genome engineering for non-therapeutic purposes. Nucleic acids encoding said variants, expression cassettes comprising said nucleic acids, vectors comprising said expression cassettes, cells or organisms, plants or animals except humans, transformed by said vectors.

Abstract: An I-MsoI homing endonuclease variant able to cleave mutant I-MsoI sites having variation at positions ±8 to ±10, a vector encoding said variant, a cell, an animal or a plant modified by said vector. Use of said I-MsoI endonuclease variant and derived products for genetic engineering, genome therapy and antiviral therapy.

Abstract: The present invention provides methods for selectively amplifying a target population of nucleic acid molecules in a population of RNA template molecules (e.g., all mRNA molecules expressed in a cell type except for the most highly expressed mRNA species). The present invention also provides a first population of oligonucleotides including the nucleic acid sequences set forth in SEQ ID NOS:1-749 and a second population of oligonucleotides including the nucleic acid sequences set forth in SEQ ID NOS:750-1498. The first population of oligonucleotides can be used, for example, to prime the synthesis of first strand cDNA molecules complementary to mRNA molecules isolated from mammalian cells without priming the synthesis of cDNA molecules complementary to ribosomal RNA molecules.

Abstract: Variants of Bacillus sp. no. 707 alpha amylase are provided that are produced more efficiently and thus more economically. Higher fermentation yields are achieved through introducing amino acid variations that promote solubility of the variant in a fermentation broth. Increased solubility allows more enzyme to remain in solution after expression in a host cell. This in turn increases the efficiency with which the expressed variant enzyme can be recovered from the fermentation broth.

Abstract: The present invention relates to methods of improving the introduction of DNA into bacterial host cells.

Abstract: The invention provides methods of removing nucleic acid contamination from reverse transcription reactions and hot-start PCR, wherein said hot-start PCR is a barrier hot-start PCR set up and/or involves a hot-start DNA polymerase, which methods comprise use of a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA. The invention further provides a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA, nucleic acids encoding said DNase and kits or compositions comprising said DNase or said nucleic acid.

Abstract: A polypeptide having a novel endoribonuclease activity; a nucleic acid encoding the polypeptide; recombinant DNA having the nucleic acid therein; a transformant transformed with the recombinant DNA; a process for producing the polypeptide comprising the steps of cultivating the transformant and collecting the polypeptide from the culture; a process for producing a digest of single-stranded RNA comprising the step of reacting the polypeptide with the single-stranded RNA; and a method for the digestion of single-stranded RNA.

Abstract: NMB1343 is an ADP-ribosylating toxin from Neisseria meningitidis. The invention provides a mutant toxin having a substitution at one or more of amino Glu-109, Glu-111 or Glu-120. The mutations(s) is/are preferably Glu to Asp. The protein of the invention preferably has reduced or eliminated ADP-ribosyltransferase and/or NAD-glycohydrolase activity relative to the wild-type protein.

Abstract: New RNases, and methods for making them, are disclosed. Each includes an amino acid sequence that is disclosed in U.S. Pat. No. 6,239,257 B1, but that is preceded by a different N-terminus residue or leader sequence.

Abstract: The present invention relates to the regulation of reproductive development, particularly to the genetic ablation of reproductive tissues in angiosperm and gymnosperm species. Reproductive-preferred promoters, regulatory elements, and cytotoxic nucleotide sequences are disclosed herein, as are constructs and methods for genetic ablation.

Abstract: The present invention features isolated polypeptides that have bactericidal and angiogenic activities. The invention features isolated polypeptides comprising amino acid sequences of RNase A ribonucleases, fragments and variants thereof, pharmaceutical compositions, and methods for treatment of a subject.

Abstract: A new RNase, and a method for making it, are disclosed. The RNase has an amino acid sequence that is disclosed in U.S. Pat. No. 6,239,257 B1, but that is preceded by a different N-terminus residue.

Abstract: A single chain homing endonuclease, comprising a first variant of I-CreI having the amino acid sequence of accession number pdb 1g9y (SEQ ID NO: 23 is residues 1-153 of pdb Ig9y) and a second variant of I-CreI variant having the amino acid sequence of accession number pdb 1g9y (SEQ ID NO: 23 is residues 1-153 of pdb Ig9y) in a single polypeptide.

Abstract: New RNases, and methods of making them, are disclosed. Each includes an amino acid sequence that is disclosed in U.S. Pat. No. 6,239,257 B1, but that is preceded by a different N-terminus residue or leader sequence.

Abstract: The present invention is a method for detecting DNA sequence differences including single nucleotide mutations or polymorphisms, one or more nucleotide insertions, and one or more nucleotide deletions. Labeled heteroduplex PCR fragments containing base mismatches are prepared. Endonuclease cleaves the heteroduplex PCR fragments both at the position containing the variation (one or more mismatched bases) and to a lesser extent, at non-variant (perfectly matched) positions. Ligation of the cleavage products with a DNA ligase corrects non-variant cleavages and thus substantially reduces background. This is then followed by a detection step in which the reaction products are detected, and the position of the sequence variations are determined.

Abstract: The present invention provides improved methods of attenuating gene expression through the phenomenon of RNA interference. The invention provides methods of synthesis of double stranded RNAs (dsRNAs) of increased potency for use as small interfering RNA (siRNA). Surprisingly and unexpectedly, siRNAs made by the methods of the invention are significantly more potent than previously available siRNAs.

Abstract: The present invention relates to means for cleaving a nucleic acid cleavage structure in a site-specific manner. Enzymes, including 5? nucleases and 3? exonucleases, are used to detect and identify nucleic acids derived from microorganisms. Methods are provided which allow for the detection and identification of bacterial and viral pathogens in a sample.

Abstract: This invention relates, in part, to newly identified polynucleotides, polypeptides, variants and derivatives thereof; processes for making the polynucleotides and the polypeptides, and their variants and derivatives; and uses of the polynucleotides, polypeptides, variants and derivatives. The invention also relates to compositions of orthogonal aminoacyl-tRNA synthetases, and pairs of orthogonal aminoacyl-tRNA synthetases, and orthogonal tRNAs that incorporate fluorinated amino acids into proteins in response to selector codons. The present invention also includes translation biochemistry methods for site-specific incorporation of fluorinated amino acids, for example, 18F- or 19F-labelled amino acids, into proteins or peptides. Such amino acids may be used as an NMR probe for characterizing protein structure, dynamics, and reactivity or for radionuclide imaging (e.g., PET). Fluorinated amino acids may also be used to stabilize proteins or peptides.

Abstract: The present invention provides humanized, chimeric and human anti-alpha-fetoprotein antibodies, fusion proteins, and fragments thereof. The antibodies, fusion proteins, and fragments thereof, as well as combinations with other suitable antibodies, are useful for the treatment and diagnosis of hepatocellular carcinoma, hepatoblastoma, germ cell tumors carcinoma and other AFP-producing tumors.

Abstract: The synthesis, biological evaluation, and molecular modeling of alkoxysuccinyl-peptidyl-haloalkyl ketones for use as proteinase K inhibitors are described. Sample preparation processes for in situ RNA or DNA analysis using such inhibitors, methods and compositions therefor are provided.

Abstract: The invention relates to the field of polynucleotide amplification. More particularly, the invention provides methods, compositions and kits for amplification of (i.e., making multiple copies of) a multiplicity of different polynucleotide template sequences using a randomly primed RNA/DNA composite primer.

Abstract: A method for obtaining improved fertility restorer lines for male sterile crop plants and a DNA construct for use in said method are disclosed. The invention relates to the simultaneous use of two different gene sequences encoding the same protein product, one being the naturally occurring wild type sequence and the other sequence being generated by modification of the wild type sequence for expression in crop plants by using codon degeneracy to avoid homology between the two sequences at the DNA and mRNA levels, each of the said sequences being placed under independent transcriptional control of different overlapping plant tissue-specific regulatory elements in the same DNA construct.

Abstract: LSD1, a homolog of nuclear amine oxidases, functions as a histone demethylase and transcriptional co-repressor. LSD1 specifically demethylates histone H3 lysine 4, which is linked to active transcription. Lysine demethylation occurs via an oxidation reaction that generates formaldehyde. Importantly, RNAi inhibition of LSD1 causes an increase in H3 lysine 4 methylation and concomitant de-repression of target genes, suggesting that LSD1 represses transcription via histone demethylation. The results thus identify a histone demethylase conserved from S. pombe to human and reveal dynamic regulation of histone methylation by both histone methylases and demethylases.

Abstract: A deployment of a toxin gene for developmental programmed cell death in bacteria is described. M. xanthus is demonstrated to have a solitary mazF gene that lacks a cotranscribed antitoxin gene. Deletion of mazF results in elimination of the obligatory cell death during development causing dramatic reduction in spore formation. Surprisingly, MrpC functions as a MazF antitoxin and a mazF transcription activator. Transcription of mrpC and mazF is negatively regulated via MrpC phosphorylation by a Ser/Thr kinase cascade. Various methods of exploiting this novel pathway are described herein.

Abstract: A polypeptide having a novel endoribonuclease activity; a nucleic acid encoding the polypeptide; recombinant DNA having the nucleic acid therein; a transformant transformed with the recombinant DNA; a process for producing the polypeptide comprising the steps of cultivating the transformant and collecting the polypeptide from the culture; a process for producing a digest of single-stranded RNA comprising the step of reacting the polypeptide with the single-stranded RNA; and a method for the digestion of single-stranded RNA.

Abstract: The present invention relates to the use of ribonucleases (RNases) in the treatment or prevention of disease.

Abstract: The present invention discloses an expression method for non-naturally-occurring amino acid-containing protein comprising: expressing in animal cells: (A) a mutant tyrosyl-tRNA synthetase that is a mutation of tyrosyl-tRNA synthetase originating in E. coli with an enhanced specificity for a non-naturally-occurring tyrosine derivative as compared with the specificity for tyrosine; (B) suppressor tRNA originating in Bacillus species, Mycoplasma species or Staphylococcus species of eubacteria and capable of binding with the tyrosine derivative in the presence of the mutant tyrosyl tRNA synthetase; and, (C) a desired protein gene that has undergone a nonsense mutation at a desired site; wherein, the tyrosine derivative is incorporated at the location of this nonsense mutation.

Abstract: Method of preparing 1-CreI meganuclease variants having a modified cleavage specificity, variants obtainable by said method and their applications either for cleaving new DNA target or for genetic engineering and genome engineering for non-therapeutic purposes. Nucleic acids encoding said variants, expression cassettes comprising said nucleic acids, vectors comprising said expression cassettes, cells or organisms, plants or animals except humans, transformed by said vectors.

Abstract: Stabilized angiogenin compositions and methods of preparing a stabilized angiogenin compositions by non-covalent immobilization on a naturally occurring substrate, such as a protein, lipid, nucleic acid or nucleotide substrate, are disclosed.

Abstract: This invention relates to cytotoxic variants of human ribonuclease 1 (RNase 1) identified through analysis of the interaction between RNase 1 and the human ribonuclease inhibitor (hRI) as defined by the three dimensional (3-D) atomic structure of the RNase1 hRI complex. Also disclosed is the 3-D structure of the hRI•RNase 1 complex and methods for designing the RNase 1 variants.

Abstract: This invention provides compositions and methods for producing translational components that expand the number of genetically encoded amino acids in eukaryotic cells. The components include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, orthogonal pairs of tRNAs/synthetases and unnatural amino acids. Proteins and methods of producing proteins with unnatural amino acids in eukaryotic cells are also provided.

Abstract: The invention relates to modified restriction enzymes capable of being used for promoting homologous recombination in organisms, in particular plants, making it possible to either target gene integration or excise unwanted DNA sequences in the genome of said organisms.

Abstract: Methods and compositions for producing siRNAs, e.g., in the form of a d-siRNA composition, from dsRNAs are provided. In the subject methods, a dsRNA is contacted with a composition that includes an activity that cleaves dsRNA into siRNAs, where the composition efficiently cleaves dsRNA into siRNAs. siRNAs produced by the subject methods find use in a variety of applications, particularly in applications where the specific reduction or silencing of a gene is desired. Also provided are kits for use in practicing the subject invention.

Abstract: This invention provides compositions and methods for producing translational components that expand the number of genetically encoded amino acids in eukaryotic cells. The components include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, orthogonal pairs of tRNAs/synthetases and unnatural amino acids. Proteins and methods of producing proteins with unnatural amino acids in eukaryotic cells are also provided.

Abstract: The invention relates to a method for producing recombinant RNase A in E. coli. Said method is characterised in that it uses a DNA sequence, which codes for an RNase A of bovine origin and which is adapted to the codon usage in E. coli. The invention relates to nucleic acid molecules, which contain a nucleic acid sequence that has been adapted to the codon usage in E. coli and to recombinant nucleic acid molecules, which contain an nucleic acid molecule of this type and permit the expression of the recombinant RNase A in E. coli.

Abstract: The present invention relates to methods and kits for the treatment of cancer. A novel drug combination comprising a ribonuclease compound and a thiazolidinedione compound has been identified as producing a synergistic cytotoxicity effect in cancer cells. Methods and kits pertaining to the co-administration of these compounds are discussed herein.

Abstract: Methods and agents for suppressing expression of a mutant allele of a gene having a polymorphism are provided. The methods of the invention provide suppression effectors such as antisense nucleic acids or ribozymes, that bind to nucleic acid regions having a polymorphism within a gene such that one allele of a gene is exclusively or preferentially suppressed. The method also provides the administration of a replacement nucleic acid, if required. The invention has the advantage that the same suppression strategy, when directed to polymorphisms, could be used to suppress, in principle, many mutations in a polymorphic gene. This is particularly relevant when large numbers of mutations within a single gene cause disease pathology.

Abstract: A method of expressing a protein having an unnatural amino acid integrated thereinto comprising expressing (A) a mutant tyrosyl tRNA synthase that is derived from Escherichia coli-origin tyrosyl tRNA synthase and has an elevated specificity for an unnatural tyrosine derivative compared with the specificity for tyrosine, (B) a suppressor tRNA originating in an eubacterium belonging to the genus Bacillus, Mycoplasma or Staphylococcus which is capable of binding to the above tyrosine derivative in the presence of the above mutant TyrRS, and (C) a desired protein gene having a nonsense mutation at a desired site in animal cells to thereby incorporate the above tyrosine derivative into the nonsense mutation site in the above protein.

Abstract: A composition for in vitro use as a culture medium or in vivo use as a pharmaceutical composition or a medical device, capable of accelerating the differentiation of stem cells into cells with a chondrocytic phenotype and of restoring the original trophism of chondrocytes, is described. The composition comprises, in combination, at least one proteolytic enzyme, at least one growth factor and at least one from a sugar, an amino acid, a vitamin factor, a vitamin, a nucleotide and a nucleoside, in a physiologically acceptable carrier or diluent. A method of differentiating stem cells in cells having a chondrocytic phenotype, the cells obtained by the method and their uses, for example in human or animal cell therapy, for example by CBMP (Cellular Based Medicinal products) are also described.

Abstract: Mutant forms of ribonuclease inhibitor are described which are rendered more resistant to oxidation while retaining affinity for both ribonuclease and angiogenin. The mutant forms have another amino acid, typically an alanine, substituted for one or more of the adjacent cysteine residues in the wild-type sequence to prevent the formation of unwanted disulfide bonds which can disrupt the effectiveness of the molecule.

Abstract: Recombinant immunotoxins containing a cytotoxic RNAse fused to an antibody or antibody fragment may be produced in mammalian cell culture. Surprisingly, immunotoxins containing a cytotoxic RNAse fused to the N-terminus of one antibody variable domain can be prepared and retain the ability to specifically bind antigen. The immunotoxins may be used in a variety of therapeutic methods for treating diseases or syndromes associated with unwanted or inappropriate cell proliferation or activation.

Abstract: The invention relates to composition or a kit containing an enzyme that is reversibly inhibited by means of a chemical modification and an enzyme which is reversibly inhibited using non-covalent binding, the use of a mixture of enzymes reversibly inhibited in such a manner for processing or multiplying polynucleotides, and a method for specifically amplifying DNA by simultaneously using both types of reversibly inhibited enzymes.

Abstract: Deficiencies in the enzymatic activity of acid sphingomyelinase (ASM) result in Niemann-Pick disease. A variety of modifications which eliminate the activity of the free thiol on the C-terminal cysteine residue of ASM all result in substantially increased specific activity of the enzyme. Methods used to alter the activity of this residue include site-directed mutagenesis to delete or alter the residue, enzymatic degradation of the ASM to remove the residue, copper-promoted dimerization of ASM (via the terminal cysteine residues) and chemical modification of the free thiol group on this residue.

Abstract: Compositions and methods of producing components of protein biosynthetic machinery that include glutamyl orthogonal tRNAs, glutamyl orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of glutamyl tRNAs/synthetases are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins using these orthogonal pairs.

Abstract: The present invention relates to a method for producing a polypeptide comprising using a signal peptide, to nucleic acid constructs comprising a first nucleotide sequence encoding the signal peptide and a second nucleotide sequence encoding a polypeptide which is foreign to the first nucleotide sequence. Furthermore, it also relates to expression vectors and host cells comprising the nuclei acid construct.