Health assessment of Black-crowned Night-herons (Nycticorax nycticorax) of the New York Harbor estuary

Blood samples from 145 Black-crowned Night-heron (Nycticorax nycticorax, BCNH) chicks (mean age 3 weeks) were taken from four island colonies (Goose (2004), Canarsie Pol (2005), Hoffman (2004 and 2005) and North Brother (2004 and 2005)) in New York Harbor in 2004 and 2005 to establish baseline health reference ranges for this species and to compare health indices of birds reared on different islands. Packed cell volume (PCV) and total solids (TS) did not differ among islands in either year. Herons raised on Hoffman Island in 2004 had lower white blood cell count (WBC), and higher activities of creatine phosphokinase (CPK), lactate dehydrogenase (LDH), aspartate amino transferase (AST), higher concentrations of potassium (K) and phosphorous (PHOS) and lower liver derived proteins (TP, prealbumin, albumin, alpha 1 globulins, alpha 2 globulins, beta globulins and gamma globulins) compared to herons from Goose and North Brother Islands. These changes suggest compromised health in chicks reared on Hoffman Island in 2004. On Hoffman in 2005, these biochemical analytes did not differ from concentrations and enzyme activities measured from birds on other islands. Although no single etiology can explain these extensive changes, it is likely that exposure to contaminants at foraging sites used by birds nesting on Hoffman and/or changes in prey availability and abundance causing birds to forage in different locations between years, led to differences measured in blood-based health indices. Avian health assessments coupled with foraging ecology serve as an excellent method for evaluating ecosystem health of the New York Harbor estuary system.     

Haplotype distribution in the GLI3 gene and their associations with growth traits in cattle

The glioma-associated oncogene family zinc finger 3 gene (GLI3) mediates in all vertebrates hedgehog (Hh) signaling that plays an essential role in the induction and patterning of numerous cell types during invertebrate and vertebrate development. In this study, a total of 6 single nucleotide polymorphisms (SNPs: 1–6) were identified by polymerase chain reaction–single stranded conformational polymorphism (PCR–SSCP) and DNA pool sequencing, including all 13 exons and 12 exon–intron boundaries within the bovine GLI3 gene. 16 haplotypes and 13 combined genotypes were revealed and the linkage disequilibrium was assessed in 708 individuals representing three main cattle breeds from China. The statistical analyses indicated that the SNP2, 3 and 4 are associated with the body weight at birth and 6 months in Nanyang cattle population (P < 0.05). No significant association was detected between 11 combined genotypes and body weight at five different ages. Our results provide evidence that polymorphisms in the GLI3 gene are associated with growth traits, and may be used for marker-assisted selection in beef cattle breeding program.     

Plasma membrane vesicles prepared from unadhered monocytes: Characterization of calcium transport and the calcium ATPase

We have purified unadhered human monocytes in sufficient quantities to prepare monocyte plasma membrane vesicles and study vesicular calcium transport. Monocytes were isolated from plateletpheresis residues by counterflow centrifugal elutriation. By combining this source and procedure, 7 x 10(8) monocytes of over 90% purity were obtained. The membranes, isolated on a sucrose step gradient, had an 18-fold enrichment in Na,K-ATPase, a 29-fold diminution of succinate dehydrogenase activity and were vesicular on transmission electron micrographs. The membrane vesicles loaded with oxalate accumulated calcium only in the presence of Mg and ATP. Calcium uptake did not occur if ATP was replaced by any of five nucleotide phosphates or if Mg was omitted. Calcium transport had a maximal velocity of 4 pmoles calcium/micrograms vesicle protein/min and a Km for calcium of 0.53 microM. The ionophore A23187 completely inhibited calcium accumulation while 5 mM sodium cyanide and 10 microM ouabain had no effect. A calcium-activated ATPase was present in the same plasma membrane vesicles. The calcium ATPase had a maximal velocity of 18.0 pmoles calcium/micrograms vesicle protein/min and a Km for calcium of 0.60 microM. Calcium-activated ATPase activity was absent if Mg was omitted or if (gamma - 32P) GTP replaced (gamma - 32P) ATP. Monocyte plasma membranes that were stripped of endogenous calmodulin by EGTA treatment showed a reduced level of calcium uptake and calcium ATPase activity. The addition of exogenous calmodulin restored the transport activity to that of unstripped monocyte plasma membranes. Thus, monocyte plasma membrane vesicles contain a highly specific, ATP-dependent calcium transport system and a calcium-ATPase with similar high calcium affinities.     

Sensitivity of pigmented mucosa and skin to freezing injury.

Pigmented areas of canine skin and oral mucosa were subjected to freezing in situ at various temperatures for the purpose of investigating possible differences in the sensitivity of epidermal cells to cold injury. Destruction of melanocytes occurred in the range of ?4 to ?7 °C, while squamous cells resisted injury even at ?20 °C. Replacement of the lost pigmented cells occurred from the normal tissue at the periphery of the injured area. The experiments suggest that selective destruction of pigmented lesions in clinical conditions may be achieved by freezing tissue to ?4 to ?7 °C or colder, but not to exceed ?20 °C in order to avoid destruction of squamous cells. The experiments also support wider use of cryosurgery for pigmented lesions of the skin and oral cavity.     

Determination of free Ca ion concentrations with an ion-selective electrode in the presence of chelating agents in comparison with calculated values.

Experimentally determined free Ca ion concentrations, measured with a Ca-selective electrode, were compared with values calculated with a computer program utilizing stability constants of the chelating agents: NTA, EDTA, and EGTA used to set the free ion concentration in the range of 10^?3 to 10^?6m. In the presence of 0.1 m KCl, 2 mm MgCl₂, 20 mm Hepes (pH 7.4), 2 mm ATP, 0.1 mm CaCl₂ (total concentration), and various ligand concentrations the measured free Ca²⁺ levels were found to be approximately six to seven times greater than the computer-derived values. Apparent stability constants for Ca-ATP, Ca-EDTA, and Ca-EGTA were determined under these experimental conditions.     

Inhibition of cobalt(II) carboxypeptidase A by ligands. Kinetic evidence for the formation of a ternary enzyme-metal-ligand complex

Saturation kinetics are observed in the inhibition of cobalt carboxypeptidase A by the chelating agent 1,10-phenanthroline. The association constant K₁ for the formation of the enzyme-metal-ligand ternary complex and k₂, the rate of breakup of the ternary complex, have been obtained. A mechanism is proposed to account for the pH profile of the reaction which, in conjunction with K₁, permits the calculation of the individual rate constants k₁, K_?1, k₂, k₃. The magnitude of the rate constant k₁ suggests that cobalt(II) in CoCPA is five-coordinate. Similar but less extensive studies on inhibition by 2,2′-bipyridyl and 8-hydroquinoline-5-sulfonic acid have also been carried out     

Wholemount histofluorescence of catecholamine-containing neurones in a hemipteran brain

Whole brains from Rhodnius prolixus treated with glyoxylic acid display catecholamine histofluorescence in eleven bilaterally distributed clusters of neurones, and in three circumscribed neuropile regions. This simple method offers resolution of some features not detected in studies on sectioned insect tissue; cephalic neurosecretory cells in chronically starved animals are histofluorescent and diverse catecholamine-handling fibres converge within the brain of this insect.     

Kinetic mechanism of the Cu(II) enzyme galactose oxidase

The steady-state kinetics of four redox reactions catalyzed by galactose oxidase have been determined. The alcohol substrate used in each case was galactose; the four oxidant substrates used were O₂, IrCl₆^2?, porphyrexide, and Fe(CN)₆^3?. With the exception of the last reagent, saturation behavior is exhibited by all substrates. Double reciprocal plots of rate data obtained varying one substrate at various concentrations of the other are intersecting for all pairs that exhibited saturation behavior. Thus, these reactions are kinetically sequential processes involving single central complexes. These complexes involve enzyme, galactose, and one molecule of oxidant, whether or not the oxidant is a one- or two-electron acceptor. This result indicates that for one-electron oxidants, an enzyme-alcohol-derived radical species may exist as a transient prior to the reaction of the second electron equivalent of oxidant. A similar substrate

transient is postulated in the reaction involving O₂. The inhibition by H₂O₂ has also been studied in detail. H₂O₂ apparently binds to the enzyme at two sites. The nature of alcohol and O₂ binding to the enzyme Cu(II) is discussed in light of these kinetic results.     

Oxidative phosphorylation by membrane vesicles from Bacillus alcalophilus

ADP and P_i-loaded membrane vesicles from l-malate-grown Bacillus alcalophilus synthesized ATP upon energization with $\text{ascorbate}\text{N,N,N′,N′-}\text{tetramethyl}\text{-p-}\text{phenylenediamine}$. ATP synthesis occurred over a range of external pH from 6.0 to 11.0, under conditions in which the total protonmotive force was as low as ?30 mV. The phosphate potentials (ΔG_p) were calculated to be 11 and 12 kcal/mol at pH 10.5 and 9.0, respectively, whereas the values in vesicles at these two pH values were quite different (?40 ± 20 mV at pH 10.5 and ?125 ± 20 mV at pH 9.0). ATP synthesis was inhibited by KCN, gramicidin, and by N,N′-dicyclohexylcarbodiimide. Inward translocation of protons, concomitant with ATP synthesis, was demonstrated using direct pH monitoring and fluorescence methods. No dependence upon the presence of Na⁺ or K⁺ was found. Thus, ATP synthesis in B. alcalophilus appears to involve a proton-translocating ATPase which functions at low .