Esterases in quail (Coturnix coturnix). Physicochemical and developmental aspects

• 1.1. Soluble esterases of digestive system organs of various developmental stages in the quail (Coturnix coturnix) were resolved by polyacrylamide gel electrophoresis into several molecular forms which were characterized as carboxylesterases, acetylesterases, cholinesterases and esterases sensitive to eserine.
• 2.2. The pI of the majority of esterasic activity in several quail and chicken tissues was observed in the range of 5.1–5.6, while the apparent molecular weight in liver extracts was 60,000.
• 3.3. The expression of the esterase multiple molecular forms was found to be both tissue- and developmental stage-specific, with electrophoretic patterns becoming more complex in number and/or staining intensity upon hatching and thereafter, especially in liver and intestine.

     

Comparative studies of phosvitin from chicken and salmon egg yolk

• 1.1. A method developed for the isolation of phosvitin from chicken egg yolk was successfully applied to the isolation of phosvitin from salmon eggs.
• 2.2. Salmon roe phosvitin is smaller in molecular size than chicken egg phosvitin.
• 3.3. Circular dichroism spectra of all phosvitins investigated displayed good similarities with spectra showing characteristics of unordered and β-sheet secondary structure.
• 4.4. The main component in the Fourier transform infrared spectra of chicken egg phosvitin is indicative of unordered conformation, whereas the Fourier infrared data of the salmon egg phosvitin are consistent with more of β-sheet structure compared to the chicken egg phosvitin.

     

Comparative study of three different forms of both native and subunit vitellogenin identified in quail plasma

• 1.1. Three different species of native vitellogenin, designated Vgα, Vgβ and Vgγ, were detected by gradient polyacrylamide gel electrophoresis (2–16%) in the plasma of untreated mature female quail and in the plasma of estrogen-induced female or male quail. The molecular weights of Vgγ, Vgβ and Vgα (in order of increasing size) were estimated to be 400,000 to 450,000 (by PAGE) or 466,000 (by analytical ultracentrifugation).
• 2.2. DEAE-cellulose chromatography resolved vitellogenin-containing plasma into peak IV, fractions of which contained Vgα and Vgβ in equal quantities, and peak III, fractions of which contained Vgα, Vgβ and Vgγ in varying proportions.
• 3.3. Peak IV fractions disssociated to give two bands (designated Vg1 and Vg2) on SDS-polyacrylamide gel electrophoresis. Pooled peak III fractions and plasma from untreated female or estrogen-induced female and male quail dissociated to give three bands (Vg1, Vg2 and Vg3). The mol. wt of Vg1, Vg2 and Vg3 were approx. 232,500, 212,000 and 194,000, respectively.
• 4.4. Peak III and peak IV vitellogenin fractions were shown to have similar amino acid compositions except that the peak III vitellogenin fraction contained twice as much cystine as the peak IV vitellogenin fraction (2.2 vs 1.1 mol%). The peak IV vitellogenin fraction contained more serine than the peak III vitellogenin fraction (11.8 vs 10.9 mol%) and more phosphorus (0.584 vs 0.516 nmol/μg protein).
• 5.5. Vgα or Vg1, in trace amounts, were detected in the plasma of untreated male quail.
• 6.6. The amino acid contents, phosphorus contents, and mol. wt of quail vitellogenins were similar to published values for other egg-laying species.

     

Gut microflora can modify fatty acid composition in liver and egg yolk lipids of laying japanese quail (Coturnix coturnix japonica)

• 1.1. The influence of the gut microflora on lipid metabolism was investigated in germ-free (GF) and conventional (CV) laying Japanese quail.
• 2.2. Serum and egg yolk cholesterol concentrations showed comparable values in both GF and CV environments.
• 3.3. The fatty acid compostion of liver lipids was modified by the presence of gut microflora. Notably, in the presence of the gut microflora, proportion of oleic acid was reduced and conversely, stearic and linoleic acids were enhanced.
• 4.4. In egg yolk lipids, the proportion of myristoleic and palmitoleic acids was significantly lowered and that of stearic acid was significantly enhanced by the presence of the gut microflora, though the difference was very small.
• 5.5. It was suggested that oleic acid could be easily either hydrogenated to stearic acid or desaturated to linoleic acid by the action of the gut microflora in Japanese quail.

     

Identification of the major annexins in Ehrlich ascites tumor cells

• 1.1. Three calcium-binding proteins have been purified from Ehrlich ascites tumor cells.
• 2.2. They were identified by amino acid sequence analysis on selected fragments obtained by tryptic digestion.
• 3.3. The proteins belong to the annexin family and were identified as annexins II, III and V.
• 4.4. Antibodies raised against the proteins were used to examine for their presence in a number of murine tissues.
• 5.5. The occurrence was found to be in reasonable accordance with earlier reports.

     

Significance of scrotal afferents within the general thermoafferent system

• 1.1.|Switching neurons, in the past referred to as a special scrotal afferent system, intergrate information not only from the scrotum, but from various body areas.
• 2.2.|They are present in male and female rats, as well as in guinea-pigs.
• 3.3.|They are present in midbrain, thalamus, hypothalamus and cortical areas.
• 4.4.|Information processing from the scrotum, is not a special system, but part of the general thermo-and noci-afferent system.
• 5.5.|Switching neurons seem to interact with behavioral, perhaps with autonomic thermoregulatory mechanisms, too.

     

The effects of cooling the egg on the respiratory movements of the hatching fowl,Gallus g. domesticus,with a note on vocalization

• 1.1. The ambient temperature of pipped eggs of the domestic fowl was reduced from 39 to 20°C for a period of 2 hr.
• 2.2. In the majority of embryos the amplitude of respiration more than doubled during the first hour, and in each embryo the frequency fell to a minimum value by the end of cooling.
• 3.3. When the eggs were rewarmed the respiratory frequency usually returned to normal within a period of 1 hr. Vocal activity was often stimulated and was accompanied by a marked increase in the amplitude of respiration.
• 4.4. These results are discussed in relation to the development of thermoregulation, and are compared with results obtained elsewhere in a similar study on the quail.

     

Phosphorylation of a 25,000-dalton protein in slow and fast muscles of the chicken

• 1.1. Protein phosphorylation in intact chicken latissimus dorsi muscle, slow anterior (ALD) and fast posterior (PLD), was compared.
• 2.2. A major difference in [³²P]phosphate incorporation was found between the ALD and PLD in a 25,000-dalton heat soluble protein.
• 3.3. The 25,000-dalton protein was purified from both the ALD and PLD.
• 4.4. The two proteins had similar amino acid composition and both contained approximately 1 mole phosphate per mole of protein.
• 5.5. The difference in their content of radioactive phosphate was determined to be due to faster turnover in the ALD.

     

The effect of water loss upon the urate,urea and ammonia content of the egg of the Japanese quail Coturnix coturnix japonica

• 1.1. The urate, urea and ammonia content of the whole egg of the Japanese quail was measured in late incubation in eggs subject to different rates of water loss.
• 2.2. High rates of water loss substantially increased egg urate content, but had little or no effect on urea or ammonia content.
• 3.3. Allopurinol, an inhibitor of urate synthesis, reduced egg urate content to low levels, but produced no effect on urea content, and a small reduction in ammonia content.
• 4.4. The urea concentration of the embryo was lower than in allantoic fluid.
• 5.5. It is concluded that urate production by the avian embryo is primarily concerned with the modification of allantoic fluid composition.

     

Contrast in adenine uptake by chicken and rabbit erythrocytes in vitro

• 1.1. Relative to rabbit erythrocytes, chicken red blood cells exhibit a much greater capacity to utilize [³H]adenine for nucleotide synthesis in vitro, even at 5°C and in the absence of added inorganic phosphate.
• 2.2. This difference is largely due to a higher concentration of phosphoribosylpyrophosphate and greater activity of adenine phosphoribosyltransferase in the avian cells. lli]3. The capacity of avian erythrocytes for utilization of guanine and hypoxanthine is several fold less than that of adenine.
• 3.4. The data are consistent with lower activity for hypoxanthine/guanine phosphoribosyltransferase than for adenine phosphoribosyltransferase in intact chicken erythrocytes.
• 4.5. The results indicate that reutilization of adenine by chicken erythrocytes may be physiologically significant.

     

Effect of adenosine on gluconeogenesis in the perfused liver of chicken and guinea-pig

• 1.1. Glucose formation from lactate by the perfused liver of 48 hr starved chickens was strongly inhibited by adenosine (Ado); the half-maximal inhibition was attained at 40 μM. This effect was paralleled by a four- to five-fold increase of ATP content as determined in freeze-clamped liver.
• 2.2. In chicken liver homogenate gluconeogenesis from precursors such as alanine, glutamate, glutamine and aspartate, which are not converted into glucose by the perfused chicken liver, proceeded at rates equal to or higher than that with lactate, being markedly inhibited by Ado.
• 3.3. In the perfused guinea-pig liver glucose synthesis with lactate, propionate, glycerol and fructose was also inhibited by Ado; however, when precursors such as pyruvate, glutamine and a mixture of lactate + pyruvate were supplied to the liver Ado did not inhibit gluconeogenesis.
• 4.4. Assay of adenine nucleotides in the perfused guinea-pig liver, stopped by freeze-clamping technique in a number of experimental variants, revealed no correlation between the rate of gluconeogenesis and the changes induced by Ado in the adenine nucleotide pool.
• 5.5. In the perfused liver of both chicken and guinea-pig Ado produced an increase of the lactate to pyruvate ratio and, in general, a diminution of the content of malate-aspartate shuttle intermediates.
• 6.6. The results are interpreted as suggesting that the inhibitory effect of Ado on hepatic gluconeogenesis is not necessarily mediated by the changes in the adenine nucleotide pool.

     

The changes in body temperature,oxygen consumption,CO2 production and muscle protein turnover rate by selection for body size in Japanese quail,Coturnix coturnix japonica

• 1.1. Body temperature, oxygen consumption, CO₂ production and muscle protein degradation rate were measured in the three quail lines selected for body size, a random bred line (RR) and two lines selected for large (LL) or small (SS) body size.
• 2.2. The body temperature at 15 weeks of age was highest for small body size line and lowest for large body size line.
• 3.3. The body temperature, oxygen consumption and CO₂ production of females were significantly higher than that of males.
• 4.4. The fractional degradation rate of muscle protein of SS, RR and LL lines were measured as 2.4, 1.6 and 1.2% per day in male, and 2.6, 1.7 and 1.4% per day in female.

     

Effects of transforming growth factor β1 and basic fibroblast growth factor on lipoprotein lipase activity in primary cultures of chicken (Gallus domesticus) adipocyte precursors

• 1.1. The effect of TGF-β and bFGF on lipoprotein lipase activity in chicken adipocyte precursors was investigated.
• 2.2. Lipoprotein lipase activity was reduced by up to 80% by incubation with TGF-β whereas bFGF had no effect.
• 3.3. Contrary to that found with the 3T3-L1 preadipocyte cell line it was not necessary for TGF-β to be present prior to the start of differentiation in order to be effective.
• 4.4. Incubation of adipocyte precursors with actinomycin D abolished the effect of TGF-β suggesting that synthesis of a protein effector is required.
• 5.5. These results indicate differences in responsiveness to TGF-β and bFGF between primary chicken adipocyte precursors and some preadipocyte cell lines.

     

Kinetic behaviour of chicken liver mitochondrial malate dehydrogenase and lactate dehydrogenase mixtures

• 1.1. In the mitochondria of chicken liver cells there is lactate dehydrogenase activity that catalyses the reduction of the oxaloacetate by the NADH.
• 2.2. The presence of lactate dehydrogenase in the malate dehydrogenase preparations causes an apparent activation in the double-reciprocal plot at high oxaloacetate concentrations that depends on the lactate dehydrogenase/malate dehydrogenase ratio in the preparation.
• 3.3. The separation of the two molecular forms of chicken liver mitochondrial malate dehydrogenase, free from lactate dehydrogenase, is described.

     

Comparative study of ovalbumins from various avian species by Con A/Sepharose chromatography

• 1.1. Ovalbumin samples from 7 chicken subspecies and other avian species were subfractionated on Con A/Sepharose column.
• 2.2. The ovalbumins from chicken subspecies were separated into 4 fractions, OA, OB, OC and OD without exceptions, although the fractional ratio (OA:OB:OC:OD) was varied from one sample to another. Similarly, turkey ovalbumin also could be separated into the 4 fractions.
• 3.3. Quail ovalbumin was tightly bound to Con A/Sepharose column and effectively eluted as a new characteristic peak (OQ) behind OD from the column at room temperature, while goose ovalbumin was totally eluted as unadsorbed fraction from the column.

     

Effects of niacin deficiency on the relative turnover rates of proteins in various tissues of Japanese quail

• 1.1. The effects of niacin deficiency on the relative turnover rates of proteins in various tissues of Japanese quail were investigated.
• 2.2. The level of liver NAD was not affected by niacin deficiency whereas the level of pectoral muscle NAD was markedly reduced.
• 3.3. In all dietary treatments the liver had the highest turnover rates of proteins, heart and brain had intermediate rates, and pectoral muscle had the lowest rates.
• 4.4. Relative turnover rates of proteins in all tissues (particularly pectoral muscle) of the niacin deficient group were significantly higher than those of pair-fed control group, although there were no significant differences in turnover rate between pair-fed control and control groups.
• 5.5. The high turnover rate of proteins in niacin deficiency was primarily attributed to enhanced degradation rate of proteins rather than enhanced synthesis rate of proteins.
• 6.6. Optical density scanning (or densitometric) of water-soluble pectoral muscle proteins separated by isoelectric focusing revealed several additional minor protein bands between major protein bands in the niacin deficient group which were more pronounced in the acidic region of the gel.
• 7.7. These results suggest that proteins with a low pI value in pectoral muscle of the niacin deficient animal are highly sensitive to protein degradation.

     

In vitro modification of cholesterol/ phospholipid ratio of enterocytes brush border membrane and its effect on l-leucine accumulation

• 1.1. Incubation of intestinal segments from chicken jejunum in liposome suspensions with different cholesterol/phospholipid ratios lead to change in the enterocytes brush border membrane cholesterol/phospholipid molar ratio.
• 2.2. The fatty acid composition, total amount and the ratios of the individual phospholipids of enterocytes brush border membrane were not affected.
• 3.3. The change in the cholesterol/phospholipid molar ratio had no effect on l-leucine accumulation in the enterocytes.

     

Gas tensions within the egg of the quail (Coturnix c. Japonica) during the closing stages of embryonic development

• 1.1. The respiratory status of the embryonic quail during the two days prior to hatching was assessed by measuring the gas tensions within the air space of the egg and of blood collected from the chorioallantois.
• 2.2. When the lungs became inflated there was a significant decrease in the pO₂ of the gas in the air space.
• 3.3. After pipping, there was a rise in the pO₂ and fall in the pCO₂ within the air space, together with corresponding changes in the blood.
• 4.4. The outer shell membrane remained intact until the onset of hatching.
• 5.5. These results were compared with those obtained by other workers using the domestic fowl.

     

Separation and properties of a “Neutral” hexosaminidase from embryonic chicken brain

• 1.1. A “neutral” hexosaminidase has been separated from other hexosaminidase forms (I and II) by DEAE-cellulose chromatography and characterized in embryonic (16-days old) and 1-day old chicken brains.
• 2.2. Its properties differ from those of the forms I and II. It has optimum activity at about pH 6.0 and can be eluted from DEAE-cellulose with 0.25 M KCl only.
• 3.3. It has no N-acetylgalactosaminidase activity and cannot be successfully detected after isoelectric focusing since it is very acidic and completely unstable below pH 5.0.
• 4.4. “Neutral” hexosaminidase is heat-stable at pH 6.0 and is inhibited by chloride.
• 5.5. These properties, very different from those of forms I and II, suggest that this “neutral” form of hexosaminidase would be very similar to known hexosaminidase C separated from other materials.
• 6.6. We have found no significant differences for the above-mentioned three forms in chick embryos (16-days old) in comparison with those from 1-day old chicken.

     

Stearoyl-CoA desaturase activity in primary culture of chicken hepatocytes. influence of insulin,glucocorticoid, fatty acids and cordycepin

• 1.1. Stearoyl-CoA desaturase (Δ9-desaturase) activity was measured in chicken primary hepatocytes, as a function of time in culture.
• 2.2. When using fasted donor animals, the desaturase activity was low at the beginning of culture and then increased steadily to a maximum value between 30 and 70 hr of culture. When hepatocyte cultures were prepared from fed animals, enzyme activity was high at the beginning of culture and maintained thereafter at similar values to those obtained in cultured hepatocytes from fasted animals after 30 hr of culture.
• 3.3. Insulin significantly enhanced enzyme activity when added to the culture medium at a 10⁻⁹M concentration, and a small stimulating effect was also observed with 10⁻⁶M dexamethasone.
• 4.4. Linoleic acid (0.5 mM) added to the culture medium as albuminic complex partly inhibited Δ9-desaturase activity.
• 5.5. Cordycepin (3' deoxyadenosine) decreased enzyme activity when present at a 3 μg/ml concentration in the culture medium.
• 6.6. Taken together, the induction of enzyme activity in culture, its impairment by cordycepin and response to insulin and linoleic acid strongly suggest that synthesis and translation of the Δ9-desaturase mRNA occur in chicken hepatocytes in primary culture, and that this cellular model may be a useful tool for further studies on Δ9-desaturase regulatory mechanisms.