The genetic control of arylesterase activity in the serum of Gotland Sheep

Esterase activity was determined in 955 serum samples of Gotland sheep. The activities showed a wide range and could be distinguished in six types. Family data were in agreement with the theory of three multiple alleles controlling the esterase activities (Lee, 1966). The following gene frequencies were estimated: Es ^a= 0.15, Es ^b= 0.75 and Es ^c= 0.10. The Es ^a allele controlled an enzym with high hydrolysis rate of α-naphthyl acetate, medium high of β-naphthyl acetate and low of indophenyl acetate. The enzym controlled by the Es ^b allele had medium high hydrolysis rate of α- and β-naphthyl acetate and relatively high of indophenyl acetate. The enzym of the Es ^c allele had low activities for all three substrates. The hydrolysis rate was increased by the addition of calcium. In starch-gel electrophoresis the esterase types could be divided in four groups due to the staining intensity of the arylesterase zone but no difference in migration rates was observed.     

Genetic control of liver esterase forms in chickens

Six regions of esterase activity designated I to VI were resolved from liver extracts of chickens by horizontal starch gel electrophoresis. These esterases were further characterized on the basis of their substrate affinities and differential responses to various inhibitors.
Genetic variation was found in esterases of region VI which appeared to be ali-esterase. Four phenotypes, A, B, AB and O, were observed. These phenotypes were shown to be controlled by one autosomal locus, designated Es-3 , with alleles Es-3 ^A, Es-3 ^B and Es-3 ^O. This locus is not closely linked to the blood group loci A and B , serum alkaline phosphatase ( Ap ), liver acid phosphatase ( Acp-2 ) and serum esterase ( Es-1 ) loci.     

Anti-Candida activity of four antifungal benzothiazoles

Abstract Anti- Candida activity of 6-amino-2- n -pentylthiobenzothiazole (I), benzylester of (6-amino-2-benzothiazolylthio)acetic acid (II) and of 3-butylthio-(1,2,4-triazolo)-2,3-benzothiazole (III) was followed and compared to that of 2-mercaptobenzothiazole (IV). I and II exhibited good activity against the C. albicans yeast form, similar to IV. They were inhibitorily active against other Candida strains, IC₅₀ values being of the order of 10⁻⁵ M, which means better activity than IV. Compound I also exhibited inhibitory activity on germ-tube formation and mycelial growth in the C. albicans strains, while II, III and IV were not active in these tests. III was the least active form of the compounds tested, IC₅₀ values being of the order of 10⁻⁴ M. All the compounds tested were highly active on a nystatin-resistant C. albicans mutant, with IC₅₀s of the order of 10⁻⁶ M−10⁻⁵ M.     

Population Dynamics of Pseudomonas syringae pv. tomato Strains on Tomato Cultivars Rio Grande and Rio Grande-Pto under Field Conditions

We examined the effects of the Pto resistance locus on the population dynamics of Pseudomonas syringae pv. tomato (Pst) strains in field experiments with the nearly isogenic tomato lines Rio Grande (RG, susceptible to Pst races 0 and 1) and Rio Grande-Pto (RG-Pto, resistant to Pst race 0, susceptible to Pst race 1). Pst strain SM78-1Sm^r (race 0) grew well under field conditions and caused ample bacterial speck disease on susceptible RG plants. In contrast, strain DC3000 failed to establish large populations when inoculated onto field grown RG plants. Mean population sizes of SM78-1Sm^r were 4–5 orders of magnitude larger on RG than RG-Pto plants indicating that RG-Pto plants were highly effective in attenuating pathogen population development. Most of the sampled leaflets from RG-Pto field plots harboured small numbers of SM78-1Sm^r. However, population sizes SM78-1Sm^r as large as 10⁵–10⁶ CFU were found on a few leaflets. Bacteria isolated from these leaflets had phenotypes characteristic of Pst race 1 strains. In growth chamber plant assays, the bacterial strains grew well and caused typical speck lesions on RG-Pto plants. The strains appeared to be race-shift mutants of SM SM78-1Sm^r. Interestingly, results from DNA hybridization experiments demonstrated that the race-shift mutants were deleted for the avirulence gene, avrPto but not for avrPtoB.     

Studies on strong and weak killer phenotypes of wine yeasts: production, activity of toxin in must, and its effect in mixed culture fermentation

R.A. MUSMANNO, T. DI MAGGIO and G. CORATZA.1999.Two different killer phenotypes were detected among K⁺ (killer) yeasts isolated from spontaneous wine fermentations using a plate bioassay. The two phenotypes differed in their degree of killer activity, and were designated as SK⁺(strong killer) and WK⁺(weak killer). Strains showing either phenotype were assayed for expression of killer activity under different growth conditions. Growth in must negatively affected expression of the killer activity of both phenotypes. The supernatant fluids from must cultures showed a lower killing effect than those from yeast phosphate dextrose broth (YPDB) cultures. The ability of the two K⁺ phenotypes to prevail on K-sensitive yeasts was studied in mixed-culture fermentation experiments. Under these conditions, only strains showing SK⁺ phenotype were able to prevail on the K-sensitive yeasts. These results suggest that the K⁺ phenotype could play a relevant role in spontaneous fermentations provided that the strain exhibits an SK⁺ phenotype, and that the latter phenotype should be preferred when selected K + strains are to be used as fermentation starters.     

Electrophoretic types of leucine aminopeptidase in sheep serum

Leucine aminopeptidase in sheep serum was studied by starch gel electrophoresis. Two phenotypes, A and B, were observed, of which the former was present in 70–90 % of all the sheep examined. These two phenotypes have been shown to be controlled by a single autosomal locus, with two alleles Lay^A and Lap^B . The Lap^A allele is dominant. The frequencies of Lap phenotypes and alleles were determined in eleven Spanish and two foreign breeds. Serum alkaline phospatase and serum leucine aminopeptidase are electrophoretically distinct.     

Esterase polymorphism in vitreous fluid of Pacific hake, Merluccius productus

Three regions of esterase activity designated I, II, and III were found on zymograms of vitreous fluids. Region I, the most anodal, exhibited two zones, one of which (the slower) was in all samples, occasionally accompanied by a second band which moved just ahead of it. Likewise, Region III, the most cathodal, exhibited two phenotypes, a common one-band and a rare two-band phenotype. Region II was occupied by one- and two-banded combinations of five different zones (a, b, c, d and e), which indicated that each band was the product of an allelic series of genes (EsII ^a, EsII ^b, etc.).
Only 11 of 15 expected phenotypes were encountered because of the rarity of alleles EsII ^a and EsII ^c. The most common allele, EsII ^b, had a frequency of 0.83 in Puget Sound hake and 0.60 in Pacific Ocean hake; this difference was highly significant. In the Pacific Ocean population agreement was good between observed and expected numbers, assuming Hardy-Weinberg equilibrium. In the Puget Sound samples deviations from expected frequencies did not appear in females or first-year fish, but deviations were significant in males 2 years and older; these deviations occurred as a marked deficiency of bd males combined with an excess of bb and dd males, particularly the latter. Inhibition tests indicated that these isozymes are B esterases. Estimations of molecular weight for the isozymes of Region II based on chromatographic and ultracentrifugation methods were 70,000 and 73,500, respectively.     

Creation and Characterization of Mitochondrial DNA-Depleted Cell Lines with "Neuronal-Like" Properties

Abstract: Mitochondrial dysfunction and attendant bioenergetic defects are increasingly recognized as playing an important role in neurodegenerative disorders. The increased attention on mitochondrial involvement points to the need for developing cell lines that have neuron-like characteristics for the genetic analysis and modeling of these diseases. We describe the creation of respiratory-deficient SH-SY5Y neuroblastoma cell lines (ρ⁰64/5) by selectively depleting mitochondrial DNA through prolonged exposure to ethidium bromide. Oxygen consumption in these cells and activities of the electron transport chain enzyme complexes I and IV that contain subunits encoded by the mitochondrial genome are eliminated. In contrast, the function of complex II, a nuclear-encoded electron transport chain component, is largely intact in these cells. The ρ⁰64/5 cells retain the ability to differentiate into cells with neuron-like phenotypes following treatment with phorbol ester or retinoic acid. Normal respiratory function is recovered by repopulation of ρ⁰64/5 cells with exogenous human platelet mitochondria. The ρ⁰64/5 cell line serves as a valuable model for the study of neurologic diseases suspected of involving mitochondrial dysfunction.     

The pha gene cluster of Rhizobium meliloti involved in pH adaptation and symbiosis encodes a novel type of K+ efflux system

The fix-2 mutant of Rhizobium meliloti affected in the invasion of alfalfa root nodules (Inf⁻/Fix⁻) is K⁺ sensitive and unable to adapt to alkaline pH in the presence of K⁺. Using directed Tn 5 mutagenesis, we delimited a 6 kb genomic region in which mutations resulted in both Inf⁻/Fix⁻ and K⁺-sensitive phenotypes. In this DNA region, seven open reading frames (ORFs) were identified and the corresponding genes were designated phaA , B , C , D , E , F and G . The putative PhaABC proteins exhibit homology to the subunits of a Na⁺/H⁺ antiporter from an alkalophilic Bacillus strain. Moreover, PhaA and PhaD also show similarity to the ND5 and ND4 subunits of the proton-pumping NADH:ubiquinone oxidoreductase respectively. Computer analysis suggests that all seven proteins are highly hydrophobic with several possible transmembrane domains. Some of these domains were confirmed by generating active alkaline phosphatase fusions. Ion transport studies on phaA mutant cells revealed a defect in K⁺ efflux at alkaline pH after the addition of a membrane-permeable amine. These results suggest that the pha genes of R. meliloti encode for a novel type of K⁺ efflux system that is involved in pH adaptation and is required for the adaptation to the altered environment inside the plant.     

Bovine plasma alkaline phosphatase activity in relation to age, J substance and β-lactoglobulin phenotypes

In Jersey cattle the plasma alkaline phosphatase (AP) activity was found to be markedly age-dependent. Among animals grouped with regard to age a marked decrease in the average plasma AP activity was shown to take place until an age of 2¹/₂ to 3 years. For the J substance and β-lactoglobulin phenotypes a dependency of age was not observed in the material studied.
Excluding animals less than 3 years old and thus eliminating variation in AP activity due to age, a significant association (P<0.01) between AP activity and J phenotypes was observed among 91 animals. The J^cs phenotype was associated with low AP activity and the j^a phenotype with high AP activity. On the average the J substance phenotypes, J^cs, J^s and j^a, exhibit an increasing plasma AP activity of 3.6, 4.2 and 6.8 King Armstrong Units, respectively, and a corresponding decrease in J substance.
In a similar comparison of the β-lactoglobulin phenotypes and plasma AP activity involving 82 lactating cows, an excess of the β-lactoglobulin BB phenotypes with high AP activity just at P = 0.05 was observed.     

Polymorphism of alkaline ribonuclease in the leucocytes of Black-and-White cattle

Studies were carried out on the polymorphism of alkaline ribonuclease in Black-and-White cattle in the north-eastern region of Poland. Leucocytes isolated from peripheral blood were used. Three phenotypes were defined, consisting of the combination of two alleles. The gene frequency was: q⁴= 0,7082, q^B= 0,2918. In calves the frequency of allele RNA-ase^B and number of RNA-ase^A/ RNA -ase^B heterozygotes were higher than in adult animals.     

Studies on the transferrins of goats. 3. Evidence for a third transferrin allele

A new allele Tf ^c in serum transferrin of goats is postulated. It was considered that serum transferrins in goats classified into six phenotypes are genetically controlled by three codominant alleles, Tf ^A, Tf ^B and Tf ^c. Frequencies of Tf ^c were low in native goats in Korea, Philipines and Thailand, and this allele is yet to be observed in other breeds of goats.     

Polymorphism of red cell acid phosphatase in dogs

In fresh red cell haemolysates from Labrador retriever dogs three acid phosphatase (Pac) phenotypes were found by starch gel electrophoresis. Family data were consistent with the theory that the Pac phenotypes are controlled by two codominant, autosomal alleles, designated Pac ^F and Pac ^S.     

Efficacy of certain disinfectants against infectious pancreatic necrosis virus

The virucidal properties of iodophor, chlorine (sodium hypochlorite), formalin, thimerosal (organic mercurial compound), malachite green, and acriflavine were tested on infectious pancreatic necrosis virus (IPNV). Iodine and chlorine showed good activity, but efficacy depended on the concentration of virus, the presence of organic matter (calf serum), and water _pH. Water hardness (0-300 mg 1⁻¹ as CaCO₃) did not affect virucidal activity. In a 5 min exposure, 4 mg 1⁻¹ available iodine inactivated 10^3.9 TCID₅₀ m1⁻¹ IPNV but 16 mg 1⁻¹ iodine were needed for inactivation of 10^6.3TCID₅₀m1⁻¹. The addition of 0-5% calf serum significantly reduced the iodine concentration and the virucidal activity. In comparison, 4 mg 1⁻¹ chlorine were needed to inactivate 10⁴⁶ TCID₅₀ m1⁻¹ IPNV in 5 min. However, the addition of 0-07 % serum greatly reduced the chlorine concentration and extended the virucidal contact time to 30 min or more. IPNV at 10^6.3 TCID₆₀ m1⁻¹ was not inactivated by exposures for 60 min to 0-2% formalin, 10 min to 0-2% thimerosal, 60 min to 5 mg 1⁻¹ malachite green, or 20 min to 500 mg 1⁻¹ acriflavine. However, acriflavine at 0-5 mg 1⁻¹ in cell culture media prevented the development of cytopathology caused by IPNV and may be useful in the treatment of the disease.     

Use-Dependent Suppression of the Nicotinic Acetylcholine Receptor Response by the Proadrenomedullin N-Terminal 20-Amino Acid Peptide in Rat Locus Coeruleus Neurons

Abstract: The effects of the proadrenomedullin N-terminal 20-amino acid peptide (PAMP) on the nicotinic acetylcholine (ACh) receptor (nAChR)-mediated inward current were investigated in neurons acutely dissociated from the rat locus coeruleus using whole-cell recording under voltage clamp. Nicotine and cytidine mimicked the ACh response, whereas the maximal response to dimethyl-phenylpiperazinium was lower in amplitude compared with that to ACh. Nicotine-induced current ( I _nic) was suppressed more effectively by mecamylamine than by hexamethonium. In addition, neither atropine nor α-bungarotoxin affected the I _nic. PAMP reversibly and noncompetitively suppressed the peak amplitude of 10⁻⁴ M I _nic. PAMP concentrations for the threshold, half-maximal inhibition, and maximal inhibition of 10⁻⁴ M I _nic were 10⁻⁸, 2.6 × 10⁻⁷, and 10⁻⁵ M , respectively. The peak amplitudes of 10⁻⁴ M I _nic elicited at 2-min intervals showed a gradual decline in the presence of 10⁻⁷ M PAMP. This decline in the I _nic was independent of the period of PAMP pretreatment. The suppression of I _nic by PAMP did not show any voltage dependency at a holding potential ( V _H) of <0 mV, although the inhibitory effect was masked by the marked inward rectification of I _nic at a V _H of 0 mV. These results suggest that PAMP could thus be a unique endogenous peptide that antagonizes the nAChR in the CNS.     

Genetic variants of phosvitin in egg yolk of the Japanese quail, Coturnix coturnix japonica

Summary. Phosvitin polymorphism in egg yolk of the Japanese quail was found by horizontal polyacrylamide gradient gel electrophoresis. Six phenotypes of yolk phosvitin designated A, B, C, AB, AC, and BC were observed in a population of 281 birds. Analysis of family data revealed that the phenotypic variation of quail yolk phosvitins was controlled by an autosomal Pv locus with three codominant alleles, Pv^a, Pv^b and Pv^c. The gene frequencies of Pv^a, Pv^b and Pv^c were 0.064, 0.824 and 0.112, respectively.     

Polymorphism of phosphoglucomutase in a German breed cattle

Haemolysates from cattle belonging to the Hochfleckvieh breed (N = 42), were studied for electrophoretic variation of phosphoglucomutase. Three phenotypes were observed which could be explained on the basis of two alleles PGMi^A and PGMi^B. The PGM'^B frequency of 0.7325 is comparatively lower than in other breeds.     

NADH diaphorase as a genetic marker for sheep red cells

Three NADH diaphorase phenotypes were observed in the red cells of sheep. Breeding data indicated that this polymorphism was under the control of two autosomal co-dominant alleles, designated Dia^F and Dia^s. Phenotype Dia F had significantly lower NADH diaphorase activity than phenotype Dia S. The frzquency of Dia^F and Dia^S was determined in 9 different breeds.     

The evidence of erythrocyte acid phosphatase by starch gel electrophoresis in the pig

Up to now no polymorphism of acid phosphatase in pigs has been reported. According to the method of Radam & Strauch (1971) 3 phenotypes of acid phosphatase, A, AB and B, were identified in haemolysates from 284 German Landrace boars. The corresponding gene frequencies¹ were found: 0.74 for P ^A and 0.26 for P^B.     

Transformation of Methanococcus maripaludis and identification of a Pst I-like restriction system

Abstract An optimized polyethylene glycol (PEG) method of transformation was developed for Methanococcus maripaludis using the pKAS102 integration vector. The frequency of transformation with 0.8 μg of plasmid and 3×10⁹ cells was 4.8×10⁻⁵ transformants cfu⁻¹, or 1.8×10⁵ transformants μg⁻¹, which was four orders of magnitude greater than with the natural transformation method. A Pst I restriction activity in M. maripaludis was also identified. Methylation of the plasmid with Pst I methylase increased the methanococcal transformation frequency at least four-fold. Also, chromosomal DNA from M. maripaludis was resistant to digestion by the Pst I endonuclease.