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Mechanistic studies of carboxypeptidase Y from Saccharomyces cerevisiae. pH and pD profiles and inactivation at low pH (pD) values.

Steady-state kinetics of carboxypeptidase Y, a proteinase from yeast, were studied by using the reaction of 4-nitrophenyl trimethylacetate as a probe. The pH profile of kcat. is sigmoidal in H2O-based buffers for the carboxypeptidase Y-catalysed hydrolysis of this ester (kcat. referring to the rate of deacylation of trimethylacetyl-carboxypeptidase Y). The corresponding pD profile in 2H2O is doubly sigmoidal, with inflexions at pD approximately 3.8 and approximately 6.8. The ionization of pKDapp. approximately 3.8 is caused by a rapid inactivation in 2H2O media by a process that is only slowly reversed on transfer to pH 7.00 phosphate buffer in H2O. The corresponding inactivation in H2O-based buffers of low pH is considerably slower (approximately 30-fold), follows a first-order rate-dependence and is very strongly pH-dependent, indicating some form of co-operative change in enzyme tertiary structure.

Carboxypeptidases↗

Global optimization of bimetallic cluster structures. II. Size-matched Ag-Pd, Ag-Au, and Pd-Pt systems.

Genetic algorithm global optimization of Ag-Pd, Ag-Au, and Pd-Pt clusters is performed. The 34- and 38-atom clusters are optimized for all compositions. The atom-atom interactions are modeled by a semiempirical potential. All three systems are characterized by a small size mismatch and a weak tendency of the larger atoms to segregate at the surface of the smaller ones. As a result, the global minimum structures exhibit a larger mixing than in Ag-Cu and Ag-Ni clusters. Polyicosahedral structures present generally favorable energetic configurations, even though they are less favorable than in the case of the size-mismatched systems. A comparison between all the systems studied here and in the previous paper (on size-mismatched systems) is presented.

Journal Article↗

Novel antitumor antibiotics, CI-940 (PD 114,720) and PD 114,721. Taxonomy, fermentation and biological activity.

The novel, broad-spectrum antitumor antibiotics, CI-940 (PD 114,720) and PD 114,721 are components of bioactive compounds produced by a novel actinomycete (ATCC 39366). The organism is unique in that whole cell analysis revealed LL-diaminopimelic acid and major amounts of arabinose. These new antibiotics exhibit potent antitumor activity in vitro and in vivo versus an array of tumors and possess strong antibacterial and antifungal activity. The above antibiotics are produced in 7,570-liter fermentors at yields of 5 approximately 8 micrograms/ml.

Actinomycetales↗

A multinational clinical validation study of PD ADEQUEST 2.0. PD ADEQUEST International Study Group.

OBJECTIVE: To clinically validate the use of the newly released kinetic modeling program, PD ADEQUEST 2.0 for Windows (Baxter Healthcare Corporation, Deerfield, IL, U.S.A.), by assessing the level of agreement between measured and modeled values of urea and creatinine clearances (CCr), glucose absorption, total drain volumes, and net ultrafiltration for all forms of peritoneal dialysis. DESIGN: A nonrandomized, multinational, prospective longitudinal study. PATIENTS: The study involved 104 adult patients [41 on continuous ambulatory peritoneal dialysis (CAPD), 63 on automated peritoneal dialysis (APD)] from 16 centers in 7 countries. All patients underwent a 4-hour peritoneal equilibration test (PET) but with varying percentage dextrose concentrations (1.5% or 2.5% dextrose) and varying fill volumes (range 1.5 - 2.5 L). Patients with a residual renal function greater than 10 mL/min were excluded, as were patients who had peritonitis within 6 weeks prior to baseline. MAIN OUTCOME MEASURES: Correlation coefficients and Bland-Altman limits of agreement were used to assess the level of agreement between measured and modeled values of weekly peritoneal urea Kt/V (pKt/V) and total Kt/V, weekly peritoneal creatinine clearance (pCCr, L/week/1.73 m2) and total CCr (L/week/1.73 m2), daily drain volume (L/day), net ultrafiltration (UF, L/day), daily peritoneal urea and creatinine mass removal (g/day), and daily peritoneal glucose absorption (g/day). Measured values were obtained from three repeat 24-hour urine and dialysate collections per patient, while modeled values were calculated using the Baxter PD ADEQUEST 2.0 program in conjunction with kinetic parameters estimated from a 4-hour PET and long-dwell exchange independent of the 24-hour collections. RESULTS: The results show there is excellent agreement between measured and modeled urea Kt/V and CCr with concordance correlation coefficients ranging from 0.83 to 0.97 among CAPD and APD patients. There was also excellent agreement between measured and modeled values of glucose absorption and total drain volumes (concordance correlations of 0.90 and 0.98, respectively). This level of agreement was further supported by a Bland-Altman analysis of individual differences, including differences between measured and modeled net UF (coefficient of clinical agreement ranged from 0.66 to 0.92). CONCLUSIONS: Data from a carefully performed PET and overnight exchange can, in combination with a scientifically and clinically validated kinetic model, provide clinicians with a powerful mathematical tool for use in CAPD and APD prescription management. Although not intended to replace actual measurements, kinetic modeling can prove useful as a means for quickly estimating approximate levels of clearance for a wide variety of alternative prescriptions. This, in turn, should speed up the process by which a physician can optimize the dose of dialysis suitable for a given patient and his/her lifestyle.

Absorption↗

Diastereoselective formation of a dipalladium(I) complex supported by a bridging tetradentate ligand, and oxidative addition of RS-H across a phosphine-bridged Pd(I)-Pd(I) bond.

The ligand CH2(P(o-C6H4NMe2)2)2 (dmapm) gives access to the first example of a dipalladium(I) complex supported by a tetradentate ligand, Pd2Cl2(mu-N,P,P,N-dmapm), which, unlike the well-known Pd2X2(mu-dppm)2 complexes (X = halide, dppm = bis(diphenylphosphino)methane), reacts with thiols to give addition of RS-H across a phosphine-bridged Pd-Pd bond.

Journal Article↗

[Study of the spatial structure of the duplex (Phn-NH(CH2)NH)pd(CCAAACA).pd(TGTTTGGC) with covalently bound 10-(2-hydroxyethyl)phenazine by in aqueous solution by 2D-(1)H-NMR and by limited molecular mechanics].

Detailed investigation of the spatial structure of duplex (Phn-NH(CH2)2NH) x pd(CCAAACA).pd(TGTTTGGC) having a covalently linked N-(2-hydroxyethyl)-phenazine in aqueous solution was continued by means of one- and two-dimensional 1H-NMR spectroscopy. Distances between the protons of the oligonucleotides as well as distances between the phenazinium and the nearest nucleotide groups protons were determined from the series of one-dimensional NOE experiments. The effective correlation time tau c determined for some proton pairs shows the phenazinium fragment to have greater internal motion than the heterocyclic bases. The deoxyribose protons coupling constants show the sugars to be in 2'-endo-conformation. The restrained molecular mechanics have yielded a possible structure of duplex in the aqueous solution fitting the experimental set of interproton distances.

Animals↗

[Study of the spatial structure of the product of intracomplex alkylation of the octanucleotide pd(TGTTTGGC) by 4-(N-methyl-N-(2-chloroethyl) amino)benzyl-5'-phosphamide derivative of the heptanucleotide pd(CCAAACA) in an aqueous solution by two-dimensional 1H-NMR spectroscopy and constrained molecular mechanics].

Using one- and two-dimensional 1H NMR spectroscopy (400 MHz), a detailed study of the spatial structure of the covalent adduct, a product of the intracomplex alkylation of octanucleotide pd[TGTTTGGC] by the 4-[N-methyl-N-(2-chloroethyl)amino]benzyl-5'-phosphamide derivative of heptanucleotide pd[CCAAACA] in aqueous solution, was continued. The distances between closely spaced protons of oligonucleotides as well as between protons of the benzylamide fragment and protons of the nearest nucleotide units were determined by measurements of the nuclear Overhauser effect. Analysis of effective correlation times for some pairs of protons of the covalent adduct showed that the values of tau c for the benzylamide fragment and nucleotide units C-1 and C-8, which directly interact with it, coincide and are smaller than those for the internal nucleotide units, suggesting the increased rigidity of the structure in the vicinity of the modification region. Using the method of constrained molecular mechanics, a probable state of the covalent adduct in solution is proposed that is in the best agreement with the experimentally obtained set of interproton distances. Problems connected with calculating the distance between a pair of protons with coincident chemical shifts and any third proton are considered.

Alkylation↗

Coupling of Alkynes on a Pd-Pd Bond to Generate an Electrophilic µ-Butenediylidene Moiety.

The first dipalladium µ-PPh(3) complex (1) was obtained by the facile loss of two CH(3)CN ligands from [Pd(2)(PPh(3))(2)(CH(3)CN)(4)](PF(6))(2) in CH(2)Cl(2). Coupling of p-tolylacetylene (p-TolC identical withCH) with 1 or its precusor afforded the µ-butenediylidene complex 2, treatment of which with CH(3)CN resulted in PPh(3) migration to give 3.

Journal Article↗

[Biochemical characterization of a new variant of glucose-6-phosphate dehydrogenase (G-6-PD) deficiency with favism: G-6-PD Bielefeld (author's transl)].

A new variant of G-6-PD with favism has been detected in a German family. After partial purification the enzyme was biochemically characterized. It revealed normal electrophoretic mobility, normal thermostability and a flat pH-activity curve. The Km-value for G-6-P was reduced, whereas this value was normal for NADP. The substrate analogues 2-desoxy G-6-P, Gal-6-p and deamino-NADP were utilized to increased rates. The biochemical data were compatible with the benign clinical course.

Adult↗

Synthesis and characterization of a novel Pd(II) complex with the condensation product of 2-(diphenylphosphino)benzaldehyde and ethyl hydrazinoacetate. Cytotoxic activity of the synthesized complex and related Pd(II) and Pt(II) complexes.

A new palladium(II) complex 1 of the condensation product of 2-(diphenylphosphino)benzaldehyde (dpba) and ethyl hydrazinoacetate (etha) was synthesized and characterized by elemental analyses, IR, and (1)H NMR spectroscopy. The bound ligand is a bidentate (PN chromophore), the remaining two coordination places being occupied by chloride ions in overall square planar geometry. The cytotoxic activity of the complex 1 and two related Pd(II) and Pt(II) complexes 2 and 3 was tested against a panel of four tumor cell lines. The activity of the complexes was similar to that of cisplatin, the most widely used metal-based antitumor drug. It is important to notice that complexes 2 and 3 were active to cisplatin-resistant U2-OS/Pt cells. Cell cycle alteration investigation, apoptotic assay and gelatin zymography in relation to invasion and metastasis of tumor cells, were performed with all the investigated complexes on Human cervix carcinoma (HeLa) cells. The results suggest that 1 has a similar effect to cisplatin, inducing apoptosis followed by arrest of cells in S phase of cell cycle, while 2 and 3 induce apoptosis without significant perturbations of cell cycle distribution.

Animals↗

Short-time sonolysis of chlorobenzene in the presence of Pd(II) salts and Pd(0).

The effect of PdSO4, PdCl2 and Pd(0) on the degradation of chlorobenzene in aqueous solution within 10 min under 200 kHz ultrasonic irradiation, was investigated. The reaction products remaining in the aqueous mixture were analyzed and quantified. The mechanism of chlorobenzene decomposition having benzene as key intermediate is discussed.

Journal Article↗

Carbon-carbon bond formation by electrophilic addition at the central carbon of the mu-eta(3)-allenyl/propargyl ligand on the Pd-Pd bond.

The mu-eta(3)-allenyl/propargyldipalladium complexes were synthesized by the reaction of the corresponding eta(1)-allenyl- or eta(1)-propargylpalladium complexes with Pd(2)(dba)(3). The X-ray diffraction analysis indicates that the dinuclear complex has a unique structure, in which two palladium, three carbon, two phosphorus, and one halogen atoms are in the same plane. These dinuclear complexes react with electrophiles, such as HCl or AcCl, at the central carbon of the mu-eta(3)-allenyl/propargyl ligand to give the mu-eta(3)-vinylcarbenedipalladium complexes. Intramolecular reaction proceeded smoothly to give cyclization products quantitatively. Addition of a catalytic amount of a palladium(0) complex dramatically accelerated the carbon-carbon bond formation. The MO calculations on the mu-eta(3)-allenyl/propargyl complexes indicated that the reaction proceeds via orbital control.

Journal Article↗

Synthesis and characterization of Pd and Pt complexes of 1,3-bis(ferrocenylchalcogeno)propanes: crystal structures of FcSe(CH2)3SeFc and [M{FcE(CH2)3E'Fc}2](PF6)2 (M = Pd, Pt; E, E' = Se, Te; Fc = [Fe(eta5-C5H5)(eta5-C5H4)]).

Reaction of a 1,3-bis(ferrocenylchalcogeno)propane, FcE(CH2)3E'Fc (L: E, E' = Se or Te; Fc = [Fe(eta5-C5H5)(eta5-C5H4)]), with a palladium(II) or platinum(II) precursor [M(NCMe)4](PF6)2 (M = Pd or Pt) in acetonitrile at room temperature led in good yield to the bis-chelate complexes [ML2](PF6)2. The structures of FcSe(CH2)3SeFc and all six complexes have been determined by X-ray crystallography. Electrochemical studies showed that electronic communication between ferrocenyl groups, absent in all three bis(ferrocenylchalcogeno)propanes, is established on complexation only for E = Se and E' = Se or Te, when the through-bond Fe...Fe distance is reduced to 13.17 A or less.

Journal Article↗

The parkin gene and its phenotype. Italian PD Genetics Study Group, French PD Genetics Study Group and the European Consortium on Genetic Susceptibility in Parkinson's Disease.

Mutations of the parkin gene on chromosome 6 cause autosomal recessive, early onset parkinsonism. This is the most frequent form of monogenic parkinsonism so far identified. The associated phenotypical spectrum encompasses early onset, levodopa-responsive parkinsonism (average onset in the early 30s in Europe), and it overlaps with dopa-responsive dystonia in cases with the earliest onset, and with clinically typical Parkinson's disease in cases with later onset. Despite clinical features, Lewy bodies are not found at autopsy in brains of patients with parkin mutations. The parkin protein possesses ubiquitin ligase activity, which is abolished by the pathogenic mutations.

Age of Onset↗