Note: Intro -- D-AMINO ACIDS: PRACTICAL METHODS AND PROTOCOLS -- D-AMINO ACIDS: PRACTICAL METHODS AND PROTOCOLS -- CONTENTS -- PREFACE -- PURIFICATION OF ALANINE RACEMASE FROM HELICOBACTER PYLORI -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- DISRUPTION OF ALANINE RACEMASE GENE OF SCHIZOSACCHAROMYCES POMBE -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- ASSAY METHOD FOR ASPARTATE RACEMASE ACTIVITY BY HPLC -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- SERINE RACEMASE OF SILKWORM, BOMBYX MORI -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- RECOMBINANT SERINE RACEMASE PREPARATION -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- D-AMINO ACID OXIDASE ACTIVITY ASSAYS -- 1. INTRODUCTION -- 2. MATERIALS -- 1. Chemicals -- 2. Enzymes -- 3. Solutions -- 4. Equipment -- 3. METHODS -- 1. Polarographic Assay with D-α-Amino Acids -- 2. Spectrophotometric Assay with D-α-Phenylglycine -- 3. Peroxidase-Coupled Assay -- 4. Determination of Steady-State Kinetic Parameters of the Reactions -- 4. RESULTS -- Activity of DAAO with D-Amino Acids as Measured with the Polarographic and Peroxidase-Coupled Assays -- Use of the Spectrophotometric Assay with D-α-Phenylglycine to Characterize the Isoforms found in DAAO Preparations -- 5. NOTES -- ACKNOWLEDGMENTS -- REFERENCES -- HISTOCHEMICAL DETECTION OF D-AMINO-ACID OXIDASE ACTIVITY IN RAT BRAIN -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- EVALUATION OF CHIRAL INVERSION OF D-AMINO ACID BY STABLE ISOTOPE METHODOLOGY -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 3.1. Resolution of DL-[2H7]leucine -- 3.1.1. Preparation of N-acetyl-DL-[2H7]leucine. , 3.1.2. Hydrolysis of N-acetyl-DL-[2H7]leucine by acylase -- 3.1.3. Acid Hydrolysis of N-acetyl-D-[2H7]leucine -- 3.2. Preparation of Sodium [4,5,5,5,6,6,6-2H7]2-oxo-4-methylpentanoate ([2H7]KIC) -- 3.3. Dose Experiments -- 3.4. Sample Preparation for GC-MS -- 3.5. GC-MS-SIM -- 3.5.1. Apparatus -- 3.5.2. Determination of Labeled and Non-Labeled Leucine Enantiomers -- 3.5.3. Determination of Labeled and Non-Labeled KIC -- 3.6. Data Analysis -- 3.6.1. Pharmacokinetic Parameters -- 3.6.2. Extent of Conversion -- 4. RESULTS -- 4.1. Synthesis of D-[2H7]leucine, L-[2H7]leucine and [2H7]KIC -- 4.2. GC-MS -- 4.2.1. Simultaneous Determination of the D- and L-enantiomers of [2H7]leucine and leucine -- 4.2.2. Determination of [2H7]KIC and KIC -- 4.3. Estimation of the Extent of Conversion -- 5. NOTES -- REFERENCES -- ENGINEERING THE SUBSTRATE SPECIFICITY OF D-AMINO ACID OXIDASE BY RATIONAL AND DIRECTED EVOLUTION METHODS -- 1. INTRODUCTION -- 1.1. Rational Design -- 1.2. Directed Evolution (Site-Saturation Mutagenesis and Error-Prone PCR) -- 2. MATERIALS -- 2.1. Molecular Biology -- 2.2. SDM (Rational Design) -- 2.3. SSM (Irrational Design of Specific Positions) -- 2.4. EP-PCR (Irrational Design) -- 2.5. Screening of Mutants Libraries -- 3. METHODS -- 3.1. SDM (Rational Design) of yeast DAAO [14,15] -- 3.2. SSM (Irrational Design of Specific Positions) of yeast DAAO -- 3.3. EP-PCR (Irrational Design) of yeast DAAO [16,17] -- 3.4. Screening of Mutants Libraries -- 4. RESULTS -- 4.1. SDM (rational design) -- 4.2. SSM (Irrational Design of Specific Positions) -- 4.3. EP-PCR (Irrational Design) -- 5. NOTES -- ACKNOWLEDGMENTS -- REFERENCES -- TRANSFORMATION OF PLANTS WITH D-AMINO ACID RESISTANCE SELECTABLE MARKERS -- 1. INTRODUCTION -- 2. MATERIALS AND METHODS -- D-Amino Acid Oxidase -- D-Serine Dehydratase -- 3. RESULTS -- 4. NOTES -- REFERENCES. , SCREENING FOR MUTANT MICE LACKING D-AMINO-ACID OXIDASE ACTIVITY -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- ASTROGLIAL EXPRESSION OF D-AMINO ACID OXIDASE: REGIONAL AND CELL-TYPE SPECIFIC EXPRESSION -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. Cell Cultures -- 2.2. Immunocytochemistry -- 2.3. Reverse Transcriptase-PCR (RT-PCR) -- 3. METHODS -- 3.1. Cell Cultures -- 3.1.1. Preparation of PDL-Coated Culture Flask -- 3.1.2. Mixed Glial Cells -- 3.1.3. Type-1 Astrocytes -- 3.1.4. Microglia -- 3.1.5. Purification of O-2A Progenitor Cells by Panning -- 3.1.6. Type-2 Astrocytes -- 3.2. Immunocytochemistry -- 3.3. RT- PCR -- 4. RESULTS -- 5. NOTES -- REFERENCES -- D-AMINO ACID OXIDASE: A VERSATILE TOOL TO STUDY THE FUNCTION OF THE GLIAL-DERIVED NEUROMODULATOR D-SERINE AT GLUTAMATERGIC SYNAPSES -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. Animals -- 2.2. Cell Cultures -- 2.2.1. Media and Chemicals -- 2.2.2. Small Material and Equipment for Cell Cultures -- 3. METHODS -- 3.1. Preparation of Hippocampal Slices for Patch-Clamp Recordings -- 3.2. Neuron-Astrocyte Cultures -- 3.2.1. Dissection of Hippocampi for Neurons Cultures -- 3.2.2. Dissociation of Hippocampi for Astrocytes Cultures -- 3.2.3. Preparation and Culture of Hippocampal Neurons -- 3.2.4. Mixed Cultures -- 3.3. Electrophysiological Recordings -- 3.3.1. Recording Apparatus -- 3.3.2. The Whole-Cell Configuration -- 3.4. Perfusion Media and Intracellular Solutions -- 3.4.1. Composition of the Extracellular Solutions -- 3.4.2. Perfusion System and Application of Test Substances -- 3.4.3. Composition of the Intracellular Solution -- 3.5. Treatment of Brain Slices or Cultures with D-Amino Acid Oxidase -- 3.6. Protocols of Electrophysiological Recordings and Brain/Cells Treatment with DAAO. , 3.6.1. Basic Considerations for Recording Synaptic Activity from Brain Slices or Cultures -- 4. RESULTS -- 4.1. Brain Slices Experiments -- 4.2. Electrophysiological Recordings Experiments in Cultures -- ACKNOWLEDGMENTS -- REFERENCES -- RESONANCE RAMAN STUDY ON CHARGE-TRANSFER COMPLEXES OF PORCINE D-AMINO ACID OXIDASE -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. Enzymes -- 2.2. Chemicals -- 3. METHOD -- 3.1. Measurements of Raman and Resonance Raman Spectra -- 3.2. Sample Preparation -- 4. RESULTS -- 5. NOTES -- REFERENCES -- IMMUNODETECTION OF D-ASPARTATE OXIDASE -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 3.1. Affinity Purification of Anti-DASPO -- 3.2. Immunoblotting Using Anti-DASPO -- 3.3. Immunoprecipitation of DASPO from Tissues -- 4. RESULTS -- 5. NOTES -- REFERENCES -- HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC DETERMINATION OF D-ASPARTATE OXIDASE ACTIVITY IN RAT TISSUES -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- CLONING OF THE HUMAN BRAIN D-ASPARTATE OXIDASE CDNA AND ITS EXPRESSION IN ESCHERICHIA COLI -- 1. INTRODUCTION -- 2. MATERIALS AND METHODS -- Isolation of Human Brain D-Aspartate Oxidase cDNA -- 1. Materials -- 2. Methods -- Construction of Expression Plasmid -- 1. Materials -- 2. Methods -- Expression and Purification of D-Aspartate Oxidase -- 1. Materials -- 2. Methods -- 3. RESULTS -- 4. NOTES -- REFERENCES -- METHODS FOR N-ACYL-D-AMINO ACID AMIDOHYDROLASE RESEARCH -- 1. INTRODUCTION -- 2. MATERIALS -- a. Screening and Assay -- b. Gene Cloning -- 3. METHODS -- 3-1. N-Acetyl-D-amino Acid Synthesis -- 3-2. Culture Condition -- a. Screening for Microorganisms Producing N-Acyl-D-Amino Acid Amidohydrolase -- b. Enzyme Production -- 3-3. Assay Methods for N-Acyl-D-Amino Acid Amidohydrolase -- a. TNBS Method -- b. D-Amino Acid Oxidase Method -- c. HPLC -- d. Ninhydrin Method. , 3-4. Gene Cloning of N-Acyl-D-Amino Acid Amidohydrolase -- a. Shotgun Cloning by the D-Amino Acid Oxidase Method -- b. Gene Amplification by PCR Using Mixed Primers Designed Based on the Conserved Amino Acid Sequence -- 4. RESULTS -- 5. NOTES -- REFERENCES -- ACTIVITY MEASUREMENT OF HYDANTOIN RACEMASE ENZYME: A KEY FOR THE PRODUCTION OF OPTICALLY PURE D-AMINO ACIDS -- 1. INTRODUCTION -- 2. MATERIALS -- 1. Substrate Synthesis -- 2. Activity Assay -- 3. METHODS -- 1. Substrate Synthesis -- 2. Activity Assay -- 4. RESULTS -- 5. NOTES -- 1. Substrate Synthesis -- 2. Activity Assay -- REFERENCES -- ENZYMES ACTING ON D-AMINO ACID AMIDES -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 1. Synthesis of D-Alanine Amide, (D-Phe)4 and other Substrates for the Characterization of the Enzymes -- 2. Screening for D-Aminopeptidase (DAP) [12,13] -- 3. Activity Measurement of DAP -- 4. Preparation of DAP -- 5. Screening (D-Phe)4 Degrading Microorganisms for Alkaline D-Peptidase (ADP) -- 6. Activity Measurement of ADP -- 7. Cloning of the adp Gene -- 4. RESULTS -- 1. Substrate Specificity and Distinguishing Features of D-Aminopeptidase (DAP) -- 2. Structure of DAP -- 3. Features of Alkaline D-Peptidase (ADP) -- 4. Substrate Specificity of ADP -- 5. Structure of ADP -- REFERENCES -- ACTIVITY ON D-TRYPTOPHAN ATTRIBUTABLE TO TRIVIAL CONFORMATIONAL CHANGE OF TRYPOTPHANASE IN HIGHLY CONCENTRATED AMMONIUM PHOSPHATE SOLUTION -- 1. INTRODUCTION -- 2. MATERIALS -- 1. Salts, Reaction Mixture and Enzyme Assay -- 2. Resolution of free D-Trp -- 3. Fluorescence and Circular Dichroism Spectrophotometers -- 3. METHODS -- 1. Effect of Different Salts on D-tryptophan Activity -- 2. Dependence on Reaction Temperature -- 3. Analysis of free D-Trp -- 4. Measurement of Fluorescence -- 5. Measurement of Circular Dichroism -- 6. D-Trp as a Competitive Inhibitor -- 4. RESULTS. , 5. NOTES.

Note: Intro -- D-AMINO ACIDS: PRACTICALMETHODS AND PROTOCOLS -- CONTENTS -- PREFACE -- AN ESSAY ON D-AMINO ACIDS:RETROSPECTION AND PERSPECTIVE -- 1. INTRODUCTION: NATURALOCCURRENCE OF D-AMINO ACIDS -- 2. METABOLISM OF D-AMINO ACIDS -- 1) Oxidative Deamination Pathway -- 2) Dehydrogenation Pathway -- 3) Racemization Pathway -- 4) Transamination Pathway -- 3. ENZYMOLOGICAL ASPECTS OFD-AMINO ACID METABOLISM -- 4. PERSPECTIVE -- REFERENCES -- DETECTION OF THE SPECIFIC D-ASPARTICACID RESIDUES IN PROTEIN -- 1. INTRODUCTION -- A. LENS: TO SEARCH THE SPECIFIC ASP RESIDUES IN LENSPROTEIN BY BIOCHEMICAL ANALYSIS -- A-1. Materials -- A-2. Methods -- A-2-1. The Identification of D-Asp Containing Protein in Lens -- A-2-2. Purification of Alpha A- and Alpha B-Crystallins from Lenses of ElderyDonors -- A-2-3. The Determination of the D-Asp Sites in Human Alpha A- orAlpha B-crystallin -- A-2-4. Method for the Determination of D/L Ratio of Amino Acids inProtein or Peptides -- A-3. Results -- A-3-1. The Specific Sites of D-Asp Residues in Alpha A- orAlpha B-Crystallins -- A-3-2. Mechanism of D-Asp and Beta-Asp Formation in Protein -- B. SKIN: DETECTION OF D-BETA-ASP-CONTAININGPROTEIN IN SKIN BY IMMUNOHISTOCHEMICAL ANALYSISUSING ANTI D-BETA-ASP-CONTAINING PROTEIN ANTIBODY -- B-1. Materials -- B-2. Methods -- B-3. Results -- C. DETECTION OFD-BETA-ASP-CONTAINING PROTEIN IN CELLS -- C-1. Materials -- C-2. Methods -- C-2-1. The Culture of the N/N1003A Cell Line -- C-2-2. Two-Dimensional Gel Electrophoresis -- C-2-3. Western Blot -- C-2-4. Image Analysis -- C-2-5. In-Gel Digestion -- C-2-6. Identification of D-beta-Asp-Containing Proteins by MALDI-TOF-MS -- C3 Results -- PROSPECTS -- REFERENCES -- DETECTION OF AMYLOID β PEPTIDES WITHL-ISOASPARTATE IN ALZHEIMER'S DISEASE -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 3-1. Peptide Synthesis [6,7]. , 3-2. Preparation of Anti-isoAsp Aβ Antibodies [8,9] -- 3-3. Specificity of Anti-isoAsp Aβ Antibodies on Dot Blot Analysis [9] -- 3-4. Immunohistochemistry [8-10] -- 4. RESULTS -- Characterization of Anti-isoaspartyl Aβ Antibodies -- Isoaspartate Formation of Aβ in Senile Plaques and Vascular Amyloid -- REFERENCES -- DETECTION OF D-AMINO ACID IN PEPTIDESBY RP-HPLC AND MASS SPECTROMETRY -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- REFERENCES -- DETERMINATION OF D-AMINO-ACIDRESIDUES IN PEPTIDES FROM ANIMALVENOM BY NMR: APPLICATION TOPLATYPUS VENOM PEPTIDE OVCNPS -- 1. INTRODUCTION -- 2. MATERIALS -- Venom Samples -- Reverse-Phase HPLC -- Synthetic OvCNPa and OvCNPb -- NMR Spectroscopy -- 3. METHODS -- Preparation of Crude Venom and Venom Gland Extracts -- Reverse-phase HPLC -- NMR Spectroscopy -- 4. RESULTS -- 5. NOTES -- REFERENCES -- PROTEIN L-ISOASPARTYL-OMETHYLTRANSFERASECATALYZES IN SITUFORMATION OF D-ASPARTATE ANDD-ISOASPARTATE IN PROTEINS -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 1. Acid Hydrolysis of Proteins and Peptides -- 2. Derivatization of Amino Acids with OPA-NAC Reagent -- 3. HPLC-Fluorescence Analysis of derivatized Amino Acids -- 4. Recommendations Regarding Blanks -- 5. Correcting for Acid Hydrolysis-Induced Racemization -- 4. RESULTS -- 5. NOTES -- REFERENCES -- PURIFICATION OF A NOVELMAMMALIAN PROTEINASE FOR D-ASPARTATECONTAININGPROTEIN, D-ASPARTYLENDOPEPTIDASE (DAEP) -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 1. Proteolytic Assay for DAEP -- 2. Purification of DAEP from Mouse Liver -- 4. RESULTS -- 5. REFERENCES -- MICRO-PURIFICATION AND STRUCTURALASSAY OF POLY-γ-GLUTAMATE,A D-AMINO ACID-CONTAINING BIOPOLYMER -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 3.1. Anion-exchange Chromatography of PGA -- 3.2. Micro-purification of PGA -- 3.3. Determination and Stereochemical Assay of PGA. , 3.4. Molecular-Size Estimation of PGA (FDNB method) -- 4. RESULTS -- 5. NOTES -- REFERENCES -- ESTIMATION OF CHRONOLOGICAL AGEUSING THE ASPARTIC ACID RACEMIZATIONMETHOD ON DENTIN SAMPLES -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS [13] -- 4. RESULTS -- Differences in the Degree of Racemization in Dentin from Different Partsof the Tooth (Upper Central Incisor) -- Estimation of Age from Whole Dentin -- 5. NOTES -- REFERENCES -- THE USE OF D-AMINO ACIDSIN PEPTIDE DESIGN -- 1. INTRODUCTION -- 2. CONFORMATIONAL CHARACTERISTICS OF D-AMINO ACIDS -- 3. β-HAIRPIN DESIGN -- 4. D-AMINO ACID AS HELIX TERMINATION SIGNALS -- 5. GUEST D-AMINO ACIDS INRIGHT-HANDED HELICAL SEGMENTS -- 6. ALTERNATING LD SEQUENCES -- 7. CONCLUSIONS -- ACKNOWLEDGMENTS -- REFERENCES -- CELLULAR APPROACH OF THE BIOGENESIS OFD-AMINO-ACID-CONTAINING PEPTIDES INEUKARYOTES: THE CRUSTACEAN MODEL -- ABSTRACT -- INTRODUCTION -- THE CRUSTACEAN HYPERGLYCEMIC HORMONE FAMILYAND THE X-ORGAN-SINUS GLAND COMPLEX -- CHARACTERIZATION OF A D-AA RESIDUE IN CRUSTACEANHYPERGLYCEMIC HORMONE SEQUENCES -- CELLULAR STUDY OF PHE3 ISOMERIZATION -- CHARACTERIZATION OF A D-AA RESIDUE INVITELLOGENESIS INHIBITING HORMONE SEQUENCE -- CELLULAR STUDY OF TRP4 ISOMERIZATION -- CONCLUDING REMARKS -- AKNOWLEDGMENTS -- REFERENCES -- DETECTION OF D-β-ASP-CONTAINING PROTEINSIN PARAFFIN-EMBEDDED OCULAR SAMPLESUSING ANTI-D-β-ASP-CONTAINING PROTEINANTIBODIES -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- Immunohistochemical Localization of D-β-Asp-Containing Proteins inAged Eyes -- Lens -- Iris-Ciliary Body -- Retina-Choroid-Sclera -- Immunohistochemical Localization of D-β-Asp-Containing Proteins inAge- and UV-Related Ocular Diseases -- Pinguecula -- Age-Related Macular Degeneration -- REFERENCES -- INDEX.

Note: Intro -- D-AMINO ACIDS: PRACTICALMETHODS AND PROTOCOLS -- CONTENTS -- PREFACE -- A SYSTEMATIC APPROACH TOTHE BRAIN D-SERINE SYSTEM -- 1. INTRODUCTION -- 2. QUANTITATIVE ASSAY OF D-SERINEIN THE CENTRAL NERVOUS SYSTEM -- 2.1. Dissection of the Brain Areas -- 2.2. GC and GC-MS Analysis -- 2.2.1. Reagents [5,6] -- 2.2.2. GC Analysis [5,6] -- 2.2.3. MS Analysis [5] -- 2.3. HPLC Analysis -- 2.3.1. Reagents [7,8] -- 2.3.2. HPLC Analysis [7,8] -- 3. RELEASE MECHANISM -- 3.1. In Vivo Microdialysis -- 3.2. Efflux of D-Serine Preloaded in the Xenopus Oocyte -- 4. BINDING SITE -- 4.1. DCK-Insensitive D-Serine Biding Site -- 4.1.1. Membrane Preparation [16] -- 4.1.2. [3H]D-Serine Binding Experiments [16] -- 5. UPTAKE ACTIVITY -- 5.1. Uptake by Synaptosomal P2 Fraction Isolated from Fresh Rat Brain -- 5.1.1. Tissue Preparation [18] -- 5.1.2. Uptake Experiments [18] -- 5.2. Uptake by C6 Glioma Cells -- 5.2.1. Culture of Rat Glioma C6BU-1 Cells [17] -- 5.2.2. Uptake experiments [17] -- 6. DIFFERENTIAL CLONING -- 6.1. RNA Finger Printing by RAP-PCR -- 6.2. Semiquantitave Co-Amplification RT-PCR [19,20] -- 7. FUNCTIONAL CLONING USING XENOPUS OOCYTES -- 7.1. Construction and Screening of Rat Cerebral Neocortex cDNA Library[21] -- 7.2. Expression Screening in Xenopus Oocytes [21] -- 7.3. Uptake Assay of D-Serine [21] -- 7.4. Semi-Quantitative RT-PCR (see also 6.3.) -- 7.5. Expression and Functional Assay of dsm-1 in Xenopus Oocyte -- 8. BEHAVIORAL STUDIES -- 8.1. Intraventricular Injection -- 8.2. Behavioral Ratings -- 8.3. Evaluation of Anti-Ataxic Effect by Falling Index -- 9. CHEMICALS -- 10. ETHICS -- ACKNOWLEDGEMENTS -- REFERENCES -- IMMUNOHISTOCHEMICAL STUDY OF D-SERINE -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 3.1. Preparation of Antigen (D-serine-glutaraldehyde-albumin conjugate)for Immunization -- 3.2. Purification of D-Serine Antibody. , 3.2.1: Making Affinity Columns -- 3.2.2: Immunoaffinity Chromatography -- 3.2.3: Checking the Crossreactivities -- 3.2.4: Further Purification if Necessary -- 3.2.5: Storage of Antibody -- 3.3. Tissue preparation -- 4. RESULTS -- 5. DISCUSSION -- 6. NOTES -- REFERENCES -- IMMUNOHISTOCHEMISTRYFOR D-SERINE IN BRAIN -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. For Purifying the Antibody and Testing the Antibody by dot Blot -- 2.2. For Using the Purified D-Serine Antibody for Immunohistochemistry -- 3. METHODS -- 3.1. Purifying and Testing the Antibody -- 3.1.1. Making Conjugates and Beads for Purification and Specificity Testing -- 3.1.2. Testing specificity by Dot Blot -- 3.2. Using the Purified Antibody for Immunohistochemistry -- 4. RESULTS -- 5. NOTES -- REFERENCES -- BRAIN SLICE PREPARATION FOREVALUATION OF THE PATHOPHYSIOLOGICALFUNCTIONS OF D-SERINE -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- NEPHROTOXICITY OF D-SERINE IN RATS -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- ACKNOWLEDGEMENTS -- REFERENCES -- PREPARATION OF A POLYCLONALANTISERUM AGAINST D-ASPARTATE -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. Preparation of Immunogen: -- 2.1.1. Conjugation of D-Asp and BSA via Glutaraldehyde -- 2.1.2. Antigen and Adjuvant Peptide Co-Adsorbed on Gold Particles -- 2.2. Production of the Polyclonal Antiserum in Rabbit -- 2.2.1. Subcutaneous Immunization -- 2.2.2. Bleeding from the Ear Artery -- 2.3. Purification of the D-Asp Antibody by Affinity Chromatography -- 2.3.1. Conjugation of D-Asp to CNBr-Activated Sepharose 4B -- 2.3.2. Purification of D-Asp Antibody -- 2.4. Dot-Blot Assay -- 2.5. Immunohistochemistry -- 3. METHODS -- 3.1. Preparation of Immunogen: -- 3.1.1. Conjugation of D-Asp and BSA via Glutaraldehyde. , 3.1.2. Antigen and adjuvant peptide co-adsorbed on gold particles -- 3.2. Production of the Polyclonal Antiserum in Rabbit -- 3.2.1. Subcutaneous Immunization -- 3.2.2. Intravenous Immunization -- 3.2.3. Bleeding from the Ear Artery -- 3.2.4. Preparation of Antiserum from Whole Blood -- 3.3. Purification of the D-Asp Antibody by Affinity Chromatography -- 3.3.1. Conjugation of D-Asp to CNBr-Activated Sepharose 4B -- 3.3.2. Purification of the D-Asp Antibody -- 3.4. Dot-blot Assay -- 3.5. Immunohistochemistry -- 4. RESULTS -- 5. NOTES -- ACKNOWLEDGEMENTS -- REFERENCES -- VESICULAR STORAGE IN AND SECRETION OFD-ASPARTATE FROM NEUROENDOCRINE CELLS -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. Reagents and Materials for Coating Coverslips with poly-L-lysine -- 2.2. Reagents for Immunohistochemistry -- 2.3. Reagents for Cell Culture -- 2.4. Reagents for Measurement of Vesicular Storage of D-Aspartate -- 2.5. Reagents for Measurement of Secretion of D-Aspartate -- 2.6. Reagents and Materials for Concentration of Acidic Amino Acids -- 2.7. Reagents and Materials for Quantitative Analysis of D-Aspartate byHPLC [see Ref. 13] -- 3. METHODS -- 3.1. Coating Coverslips with Poly-L-lysine -- 3.2. Immunohistochemistry -- 3.3. Cell Culture -- 3.4. Vesicular Storage of D-Aspartate -- 3.5. Exocytosis of D-Aspartate -- 3.6. Concentration of Acidic Amino Acids -- 3.7. Quantitative Analysis of D-Aspartate by HPLC -- 4. RESULTS -- 5. NOTE -- REFERENCES -- D-ASPARTATE IN FROG HARDERIAN GLAND -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- THE ENDOCRINE REGULATION OF THE TESTISIS MEDIATED BY D-ASPARTIC ACID:A STUDY IN THE LIZARD PODARCIS S. SICULA -- 1. INTRODUCTION -- 2. MATERIALS -- Animals -- In Vivo Experimental Procedures on the Lizards: -- Sex Steroid Assay in Plasma and Testis -- Specific Determination of D-Aspartic Acid. , Biosynthesis of D-Asp by a Racemase Activity -- 3. METHODS -- Administration of D-Asp into Lizards -- Sex Steroid Assay in Plasma and Testis -- Specific Determination of D-Aspartic Acid -- Biosynthesis of D-Asp by a Racemase Activity -- Statistical Analysis -- 4. RESULTS -- 5. NOTES -- REFERENCES -- D-ASPARTIC ACID IN THE NERVOUS SYSTEM OFA MOLLUSC: AN IMMUNOHISTOCHEMICALPROTOCOL -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- ACKNOWLEDGEMENTS -- REFERENCES -- ANALYSIS, DISTRIBUTION, ANDPHYSIOLOGICAL FUNCTIONS OF FREED-ALANINE IN AQUATIC INVERTEBRATES -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 1. Analyses of D- and L-Amino Acids in Invertebrate Tissues -- 2. Distribution of D-Alanine in Aquatic Invertebrate -- 3. Physiological Function of D-Alanine as a Potent Osmolyte inInvertebrate Tissues -- 4. Other Possible Physiological Functions of Free D-Alanine in AquaticInvertebrates -- 5. Biosynthesis of D-Alanine in Aquatic Invertebrates -- 6. Utilization of D-Amino Acids in Fish -- 5. CONCLUSION -- REFERENCES -- D-GLUTAMIC ACID INDUCED MUSCLECONTRACTION IN SILKWORM LARVA -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- PRIMARY STUDY OF D-AMINOACID ACCUMULATION SYSTEM -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- D-AMINO ACIDS AND IMMUNE RESPONSE -- 1. INTRODUCTION -- 2. IMMUNOGENICITY -- 3. ANTIGENIC SPECIFICITY AT THE B CELL LEVEL -- 4. PREFERENTIAL FORMATION OFANTIBODIES TO D-AMINO ACID SEQUENCES -- 5. THYMUS INDEPENDENCE -- 6. T-CELL IMMUNITY -- 7. RETRO-INVERSO PEPTIDES -- 8. AN L-POLYMER, BUT NOT ITSD-ENANTIOMER, AS A DRUG AGAINST MS -- 9. CONCLUDING REMARKS -- REFERENCES. , CONSIDERATIONS FOR THE USE OF 'RAPID'CELL SUPERFUSION AND VOLTAGE CLAMP TOINVESTIGATE THE ROLE OF RARE AMINOACIDS IN SYNAPTIC TRANSMISSION -- 1. INTRODUCTION AND THEORETICAL CONSIDERATIONS -- 2. MATERIALS -- 3. METHODS -- Whole-Cell Voltage Clamp -- Application of Amino Acids -- 4. RESULTS -- 5. NOTES -- Synaptic Complexity -- Application of Amino Acids -- AKNOWLEDGEMENTS -- REFERENCES -- ARE D-AMINO ACIDSPREVALENT AMONG EUKARYOTES? -- 1. FREE AMINO ACIDS -- 2. PEPTIDE AMINO ACIDS -- REFERENCES -- NUTRITIONAL EVALUATION OF D-AMINO ACIDS -- 1. INTRODUCTION -- 2. METHODS -- Growth Assay in Mice -- 3. RESULTS -- D-Alanine -- D-Arginine -- D-Aspartic Acid -- D-Cysteine -- D-Cystine -- D-Histidine -- Lanthionine (LAN) Isomers -- D-Lysine and Lysinoalanine (LAL) Isomers -- D-Methionine -- D-Phenylalanine -- D-Proline -- Selenomethionine Isomers -- D-Serine -- D-Threonine -- D-Tryptophan -- D-Tyrosine -- D-Valine -- 4. NOTES -- ACKNOWLEDGMENTS -- REFERENCES -- D-AMINO ACIDS IN FOOD -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 3.1. GC -- 3.2. HPLC -- 3.3. CE -- 3.4. Sensor and Biosensor -- 3.5. Yoghurt Production -- 3.6. Yoghurt Sample Preparation for Amino Acid Analysis -- 3.7. Cheese Sample Preparation for Amino Acid Analysis -- 3.8. In Vivo Experiments -- 3.9. Serum Sample Preparation for Amino Acid Analysis -- 4. RESULTS AND DISCUSSION -- 4.1. Methods for Chiral Separation of D,L-Amino Acids -- 4.2. D-Amino Acids formed in Technological Processes (Heat Treatments) -- 4.3. D-Amino Acids as Markers of Contamination in Unfermented Foods -- 4.4. D-Amino Acids in Fermented Foods -- 4.4.1. Cheese -- 4.4.2. Yoghurt -- 4.5. Metabolic Fate of Dietary D-Amino Acids -- 5. CONCLUSIONS -- ACKNOWLEDGEMENTS -- REFERENCES -- D-AMINO ACID DETERMINATION IN FOODS,BEVERAGES, AND BIOLOGICAL SAMPLES -- 1. INTRODUCTION -- 2. DETERMINATION OF NATIVE AMINO ACIDS. , 3. DERIVATIZATION METHODS FOR HPLC ANALYSIS.

Note: Intro -- D-AMINO ACIDS: PRACTICAL METHODS AND PROTOCOLS -- CONTENTS -- PREFACE -- CHROMATOGRAPHIC DETERMINATION OF FREE D- AND L-AMINO ACIDS IN MARINE ALGAE -- 1. INTRODUCTION -- 2. MATERIALS AND METHODS -- Materials -- Instruments -- Reagents -- Methods -- Sources of Algae -- Sample Preparation -- Derivatization of Amino Acids -- HPLC Analysis -- Calculations -- Identification of D-Aspartate and D-Alanine Peaks -- 3. RESULTS AND DISCUSSION -- Chromatograms of Standard L- and D-Amino Acids and Sample Extract -- Applications -- REFERENCES -- DETERMINATION OF ACIDIC D-AMINO ACIDS AND THEIR N-METHYL DERIVATIVES IN BIVALVES AND OTHER MOLLUSKS -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. Animals -- 2.2. Chemicals and Others -- 3. METHODS -- 3.1. Determination of Acidic D-Amino Acids Together with Neutral and Basic D-Amino Acids [4] -- 3.1.1. Preparation of Tissue Extracts -- 3.1.2. Derivatization -- 3.1.3. HPLC -- 3.2. Determination of Acidic N-Methyl D-Amino Acids [6,7,8] -- 3.2.1. Preparation of Tissue Extracts -- 3.2.2. Pretreatment and Derivatization -- 3.2.3. HPLC Analysis -- 4. RESULTS -- 4.1. Determination of D-Aspartate and D-Glutamate together with Neutral and Basic D-Amino Acids [4] -- 4.2. Determination of N-Methyl-D-Aspartate and N-Methyl-D-Glutamate [6,7,8] -- 5. NOTE -- REFERENCES -- PURIFICATION AND DETERMINATION OF N-METHYL-D-ASPARTIC ACID (NMDA) IN BIOLOGICAL TISSUES -- 1. INTRODUCTION -- 2. CHEMICALS AND REAGENTS -- 3. METHODS FOR THE PURIFICATION AND DETERMINATION OF NMDA -- 3.1. Purification -- 3.2. Determination of NMDA by HPLC -- 3.3. Thin-Layer Chromatography and Enzymatic Method for the Determination of NMDA -- 4. RESULTS -- 5. NOTES -- REFERENCES -- A SENSITIVE ONE STEP HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF N-METHYL-(D AND L)-ASPARTATE, N-METHYL- (D AND L)-GLUTAMATE AND (D AND L)- ASPARTATE IN BIOLOGICAL TISSUES. , 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- ACKNOWLEDGMENTS -- REFERENCES -- METHODS FOR TOTAL DETERMINATION OF D-ASPARTIC ACID, D-GLUTAMIC ACID AND N-METHYL-D-ASPARTIC ACID AND FOR THE SPECIFIC DETERMINATION OF D-ASPARTIC ACID -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 3.1. Extraction and Purification of free Amino Acids from Biological Tissues -- 3.2. Colorimetric Method for total Determination of D-Asp, D-Glu and NMDA Based on the Measurement of the α-Ketoacids -- 3.3. Colorimetric Method for total Determination of D-Asp, D-Glu and NMDA based on the H2O2 Measurement -- 3.4. Fluorimetric Method for Determination of D-Asp, D-Glu and NMDA Based on the Oxidized Tyramine Measurement -- 3.5. Enzymatic-HPLC Method for the Specific Determination of D-Asp -- 4. RESULTS AND NOTES -- REFERENCES -- AN IMPROVED HPLC METHOD FOR DETERMINATION AND QUANTIFICATION OF D- AND L-ASPARTIC ACID -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- ACKNOWLEDGMENTS -- REFERENCES -- DETERMINATION OF FREE D-ASPARTATE AND D-SERINE IN RAT BRAIN AND PERIPHERY BY GAS CHROMATOGRAPHY AND HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. Materials for GC -- 2.2. Materials for HPLC -- 3. METHODS -- 3.1. Animals and Tissue Preparation for GC and HPLC Analysis -- 3.2. Derivatization Procedure for GC -- 3.3. GC Conditions -- 3.4. Derivatization Procedure for HPLC -- 3.5. HPLC Conditions -- 4. CALCULATIONS -- 5. RESULTS -- 6. NOTES -- 6.1. Notes for GC -- 6.2. Notes for HPLC -- REFERENCES -- AN LC METHOD FOR THE ANALYSIS OF D-AMINO ACIDS -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- I. Separation -- One-Column System - Chiral Column for Separation of Single D- and L-Amino Acids -- Coupled-column - System for Separation of D- and L- Amino Acids as Constituents of a Mixture or a Protein. , II. Hydrolysis -- 4. RESULTS -- 5. NOTES -- REFERENCES -- SENSITIVE DETERMINATION OF D-AMINO ACIDS IN RAT BRAIN BY HPLC WITH A PIRKLE-TYPE CHIRAL COLUMN, FOLLOWING PRECOLUMN FLUORESCENCE DERIVATIZATION WITH NBD-F -- 1. INTRODUCTION -- 2. MATERIALS -- 2-1. D-Aspartate in the Pineal Gland -- 2-2. D-Serine in the Microdialysate from Rat Brain -- 3. METHODS -- 3-1. D-Aspartate in the Pineal Gland -- 3-2. D-Serine in the Microdialysate from Rat Brain -- 4. RESULTS -- 4-1. D-Aspartate in the Pineal Gland -- 4-2. D-Serine in the Microdialysate from Rat Brain -- 5. NOTES -- REFERENCES -- DETERMINATION OF SMALL AMOUNTS OF D-AMINO ACIDS IN MAMMALS USING COLUMN-SWITCHING HPLC -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. Reagents and Equipment for Tissue Treatment -- 2.2. Reagents and Equipment for NBD Derivatization of Amino Acids -- 2.3. HPLC System -- 3. METHODS -- 3.1. Preparation of Tissue Samples -- 3.2. Fluorescence Derivatization of Amino Acids -- 3.3. HPLC Determination of Amino Acids -- 4. RESULTS -- 4.1. Determination of D-Ala in the Rat Tissues and Physiological Fluids [11] -- 4.2. Determination of D-Leu and D-Pro in the Brains of ddY/DAO+ Mice and ddY/DAO- Mice [12,13] -- 5. CONCLUSIONS -- 6. NOTES -- REFERENCES -- SENSITIVE DETERMINATION OF AMINO ACID ENANTIOMERS BY CE/LIF -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- METHODS OF DETECTION FOR AMINO ACID ENANTIOMERS IN THE VERTEBRATE RETINA -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- REFERENCES -- DETERMINATION OF NG-NITRO-ARGININE ENANTIOMERS IN RAT PLASMA BY CAPILLARY ELECTROCHROMATOGRAPHY WITH A CHIRAL MOBILE PHASE -- 1. INTRODUCTION -- 2. MATERIALS -- 3. METHODS -- 4. RESULTS -- 5. NOTES -- REFERENCES -- DETERMINATION OF D-ENANTIOMERS IN AMINO ACID MIXTURES AND PEPTIDES SUBJECTED TO SEQUENCING -- 1. INTRODUCTION -- 2. MATERIALS. , 3. METHODS -- 3.1. Quantitative Determination of Amino acid Chirality in Protein Hydrolysates -- 3.2. Determination of Amino acid Chirality During Subtractive Edman Degradation of Peptides -- 4. RESULTS -- 5. NOTES -- REFERENCES -- QUANTITATIVE DETERMINATION OF THE PROTEIN OF BACTERIAL ORIGIN BASED ON D-AMINO ACID CONTENTS -- 1. INTRODUCTION -- 2. MATERIAL AND METHOD -- 2.1. Methodology for the Animal Experiment -- 2.2. Preparation of Samples for Chemical Analysis -- 2.3. Chemical Analysis of Samples -- 3. RESULTS AND DISCUSSION -- 4. CONCLUSIONS -- REFERENCES -- METHODS FOR DETECTION OF D-AMINO ACIDS USING D-AMINO ACID OXIDASE -- 1. INTRODUCTION -- 2. MATERIALS -- 2.1. Materials for the Oxygen Consumption Assay -- 2.2. Materials for the Spectrophotometric Assays -- 2.3. Materials for the Biosensor -- 2.4. Materials for in vivo Determination of D-Serine -- 3. METHODS -- 3.1. Determination of Oxygen Consumption -- 3.2. Methods for the Spectrophotometric Assays -- 3.2.1. Determination of -Keto Acid Production: Direct Method -- 3.2.2. Determination of -Keto Acid Production: Indirect Method with DNP -- 3.2.3. Determination of Hydrogen Peroxide Production: Indirect Method with o-Dianisidine -- 3.2.4. Determination of Hydrogen Peroxide Production: Indirect Method with 4-Aminoantipyrine -- 3.2.5. Determination of Ammonia Production -- 3.3. Methods for the Biosensor -- 3.4. Determination of D-Serine Released from Astrocytes and C6 Glioma Cells -- 4. RESULTS -- 4.1. Oxygen Consumption Assay -- 4.2. Spectrophotometric Assays -- 4.3. RgDAAO Biosensor -- 4.4. Enzyme-linked Assay to Monitor D-Serine Release from Cultured Cells -- 5. NOTES -- 5.1. Oxygen Consumption Assay -- 5.2. Spectrophotometric Assays -- 5.3. RgDAAO Biosensor -- 5.4. Enzyme-linked Assay to Monitor D-Serine Release from Cultured Cells -- ACKNOWLEDGMENTS -- REFERENCES -- INDEX.