Serum albumin polymorphism and its relationship to economic traits in crossbred cattle

The serum albumin genotypes of 65 Jersey × Hariana (F₁), 75 Holstein Friesian × Hariana (F₁) and 47 Brown Swiss × Hariana (F₁) crossbred cows were determined by starch gel electrophoresis. Two albumin alleles Alb^F and Alb^s , but only either as Alb^F homozygotes or Alb^Fs heterozygotes, were observed amongst these animals. There were no Alb^S homozygotes or other genotypes. Highly significant relationships between albumin genotypes and both birth weights and first-lactation milk yields of these cows were observed. The Alb^F allele was associated with increased milk yield and greater birth weights.     

Linkage between the loci for serum albumin and vitamin D binding protein (GC) in the Japanese quail

Vitamin D binding protein ( GC ) and serum protease inhibitor ( PI ) have been added to genetic markers in the Japanese quail. Both loci were controlled by autosomal codominant alleles named GC^A, GC^B and PI^A, PI^B, PI^C, respectively. Close linkage between the loci for serum albumin ( ALB ) and GC protein is reported. Two recombinants were observed in 145 informative offspring of 14 families. The recombination frequency between the loci was estimated as 0.014±0.006. Thus, GC was assigned to linkage group II in the Japanese quail. No signs of linkage were observed among the loci for the ALB-GC complex, PI. serum prealbumin 2 ( PA2 ), phosphoglucose isomerase ( PG1 ), 6-phosphogluconate dehydrogenase ( PGD ) and esterase-D ( ESD ).     

Transport of Zinc-65 at the Blood-Brain Barrier During Short Cerebrovascular Perfusion in the Rat: Its Enhancement by Histidine

Abstract: Zinc-65 transport into different regions of rat brain has been measured during short vascular perfusion of one cerebral hemisphere with an oxygenated HEPES-containing physiological saline at pH 7.40. The [Zn²⁺] was buffered with either bovine serum albumin or histidine. In each case uptake was linear with time up to 90 s. ⁶⁵Zn flux into brain in the presence of albumin followed Michaelis-Menten kinetics and for parietal cortex had a K _m of 16 n M and a V _max of 44 nmol/kg/min. Increasing concentrations of l -histidine enhanced ⁶⁵Zn flux into brain at [Zn²⁺] values between 1 and 1,000 n M . The combined effect of [histidine] and [Zn²⁺] was best accounted for by a function of [ZnHis⁺], i.e., flux = 64.4 · [ZnHis⁺]/(390 + [ZnHis⁺]) + 0.00378 · [ZnHis⁺], with concentrations being nanomolar. d -Histidine had an influence similar to that of l -histidine. ⁶⁵Zn flux in the presence of 100 µ M l -histidine was not affected by either 500 µ M l -arginine or 500 µ M l -phenylalanine. The results indicate specific transport of Zn²⁺ across the plasma membranes of brain endothelium. The enhancement due to histidine has been attributed to diffusion of ZnHis⁺ across unstirred layers "ferrying" zinc to and from transport sites.     

Normal ranges of some blood chemistry parameters in adult farmed Atlantic salmon, Salmo salar

Blood samples from healthy adult Atlantic salmon fed an optimal diet in net sea pens were collected at intervals from October to May. Haematological determinations and biochemical serum analyses were carried out on 20 fish in each of seven samples. The ranges of haemato-logical values for sample means were: haematocrit 44–49%, haemoglobin 8.9–10.4 g 100ml⁻¹, red blood cell count 0.85–1.10 × 10¹² l⁻¹, MCV 441–553 × 10⁻¹⁵ 1, MCH 94–106 × 10⁻⁶ g, MCHC 19.4–21.7 g 100ml⁻¹ and leucocrit 0.43–0.96%. The ranges of enzyme activities in serum, for sample means, were: alkaline phosphatase 647–988Ul⁻¹, aspartate aminotrans-ferase 202–351 Ul⁻¹ and alanine aminotransferase 4–8 Ul⁻¹. The ranges of the other parameters analyzed in serum were: total protein 41.6–56.6 gl⁻¹, albumin 18.3–24.3 gl⁻¹, albumin/total protein ratio 39.3–44.0%, creatinine 26–46 μmol, triglycerides 2.53–4.98 μmol and cholesterol 9.3–12.8 μmol. These values are considered to be the normal ranges in healthy fish. Variations due to seasonal changes, and the clinical significance of the selected parameters, are discussed. Data showing the reproducibility of the biochemical analyses in serum are presented.     

Blood protein polymorphism in the one-humped camel (Camelus dromedarius) in the Sudan

The blood protein polymorphism of serum albumin, haptoglobin, transferrin, ceruloplasmin and haemoglobin have been studied in 135 samples from one-humped Arabian camel (Camelus dromedarius) of the Sudan by starch gel electrophoresis. Only the serum albumin and haptoglobin systems exhibited polymorphism with the estimated frequencies of 0.0222, 0.2227 and 0.7773 for Alb^v, Hp¹Hp⁰respectively. The frequency of Hp was 0.0325. No electrophoretic variant was observed at transferrin, ceruloplasmin and haemoglobin loci in the camel. The activity of the ceruloplasmin of the camel sera was weak.     

Determination of xanthoxin by immunoassay

2- Cis (-)xanthoxin (XA) was linked to bovine serum albumin through a Schiff's base and the adduct stabilized by sodium borohydride reduction. The conjugate (molar coupling ratio: 3 mol XA per mol protein) was highly immunogenic in rabbits. Antisera contained antibodies binding XA with high affinity (K_a= 1.8 × 10⁸ M ⁻¹). [³H]-XA (2.2 × 10¹⁴ Bq mol⁻¹) was synthesized by oxidation of [³H]-XA alcohol with MnO₂ and used to set up a radioimmunoassay [RIA, detection limit, 1 pmol; measuring range, 1 to 200 pmol (0.3 to 60 ng) XA]. The sera were also suitable for enzyme-linked-immunosorbent assay (ELISA) using XA-alkaline phosphatase conjugates. The technique was more sensitive [detection limit, 0.1 pmol; measuring range, 0.1 to 50 pmol (0.05 to 15 ng) XA] than the radioimmunoassay, but less precise.     

Potentiation of 45Ca Uptake and Acute Toxicity Mediated by the N-Methyl-D-Aspartate Receptor: The Effect of Metal Binding Agents and Transition Metal Ions

Abstract: The activities mediated by the N -methyl-D-aspartate (NMDA) receptor were studied in cultured rat cerebellar granule cells. Micromolar concentrations of the metal binding compounds, EDTA, cysteine, and histidine, as well as serum albumin strongly potentiated receptor activity in the presence of millimolar concentrations of Ca²⁺ and Mg²⁺. The findings indicated that these agents remove an endogenous metal, probably Zn²⁺, which attenuates NMDA receptor-mediated ⁴⁵Ca uptake and toxicity. Several added metal ions were therefore tested at low micromolar concentrations. Zn²⁺ was found to be the most potent inhibitor of NMDA-induced ⁴⁵Ca uptake, followed by Cu²⁺ and Fe²⁺. Co²⁺, Cd²⁺, Fe³⁺, and AI³⁺ had no significant effect, whereas Ni²⁺ potentiated the ⁴⁵Ca uptake but inhibited at much higher concentrations. The potentiating agents that remove the endogenous metal had a particularly dramatic effect in the presence of Mg²⁺, the voltage-dependent suppressor of the NMDA receptor. Mg²⁺ also played an important role in the inhibitory effect of added Zn²⁺. Much lower concentrations of Zn²⁺ were needed to achieve inhibition of NMDA-induced ⁴⁵Ca uptake in the presence of Mg²⁺. Under a variety of conditions, a very good correlation was found between NMDA receptor-mediated ⁴⁵Ca uptake and the magnitude of acute neurotoxicity.     

Stimulation of Na+-Dependent, Veratridine-Sensitive Synaptosomal Amino Acid Uptakes by Serum Albumin

Abstract: Bovine serum albumin (BSA) is shown to stimulate selectively the synaptosomal uptakes of those amino acids that are dependent on external Na⁺ and that are inhibited by veratridine. Thus, the stimulation can be seen in the case of aspartic acid, glutamic acid, glycine, proline, and γ-aminobutyric acid, but not with serine and threonine. Further, studies on the interaction of veratridine, valinomycin, and BSA on the uptake of proline suggest that the primary action of the albumin is to increase the influx of proline. Such an action could result as a consequence of stabilization of the Na⁺ gradient by increased endogenous levels of ATP. Intrasynaptosomal ATP was increased in the presence of BSA but significantly decreased by veratridine.     

Callus formation from cotyledon protoplasts of Pinus oocarpa and Pinus patula

Protoplasts were isolated from cotyledons of 11-day-old seedlings of Pinus oocarpa and P. patula ssp. tecunumanii . The best enzyme combination was Cellulase R10 + Pectolyase Y-23, associated with bovine serum albumin. When cultured at a low density [1.25 × 10³ to 5 × 10³ protoplast (ml)⁻¹] in a liquid medium, the cells divided. The medium contained glutamine and casein hydrolysate as nitrogen sources, and glucose as osmoticum. Rate of division was increased by supplementing the medium with l -ornithine, putrescine and spermidine. However, the rate remained low, with an absolute division frequency of ca 1%. Dilution allowed colony proliferation and fragmentation, leading to the formation of numerous microcalli that could be transferred to various solid media for further growth.     

Early Signals in Serum-Induced Increases in Ouabain-Sensitive Na+-K+ Pump Activity and in Glucose Transport in Rat Skeletal Muscle Are Amiloride-Sensitive

Abstract: The acute effects of serum on sodium-potassium (Na⁺-K⁺) pump activity and glucose uptake in cultured rat skeletal muscle were studied. Addition of serum to myo-tubes in phosphate-buffered saline caused Na⁺-K⁺ pump activity (as measured by changes in the ouabain-sensitive component of both membrane potential and ⁸⁶Rb uptake) to increase, with peak effects obtained after 30 min. The effect was blocked completely by treatment with amiloride, but not by tetrodotoxin, which blocks voltage-dependent Na⁺ channels. On transfer of myotubes to Na⁺-free, choline buffer, resting Na⁺-K⁺ pump activity decreased to about 10% of that in phosphate-buffered saline. Addition of regular serum, but not Na⁺-free serum, caused Na⁺-K⁺ pump activity to increase slightly. Similar results were obtained with serum on glucose uptake, the peak effect being reached within 15 min. Stimulation of glucose uptake by serum was partially reduced by amiloride and was not altered by tetrodotoxin. Removal of external Na⁺ also eliminated serum effects on glucose uptake. The results demonstrate that there are similar signals involving Na⁺-H⁺ exchange for serum-induced increases in Na⁺-K⁺ pump activity and glucose transport. The lack of complete blockade of serum-induced elevation of glucose transport suggests an additional, as yet undefined, intracellular signal for stimulation of this transport system.     

Further contribution to the H blood group system in pigs

The haemolytic reagent allowing direct serological detection of He homozygous pigs was obtained by the immunization of a Landrace sow. Another monospecific reagent prepared from immune serum of a Miniature pig made possible the evidence for a new factor of the H system - He. This factor is genetically controlled by the allele H ^be. Its frequency in Miniature pigs was 0.099.
The H system with alleles H ¹=H^a, H²= H^b, H³= H^ab, H⁴=H^cd, H⁵= H^bd, H⁶=H^be and H⁷= H- continues to be a genetically open system.     

Comparative Detection of Calcium Fluctuations in Single Female Sex Cells of Tobacco to Distinguish Calcium Signals Triggered by in vitro Fertilization

Double fertilization is a key process of sexual reproduction in higher plants. The role of calcium in the activation of female sex cells through fertilization has recently received a great deal of attention. The establishment of a Ca²⁺-imaging technique for living, single, female sex cells is a difficult but necessary prerequisite for evaluating the role of Ca²⁺ in the transduction of external stimuli, including the fusion with the sperm cell, to internal cellular processes. The present study describes the use of Fluo-3 for reporting the Ca²⁺ signal in isolated, single, female sex cells, egg cells and central cells, of tobacco plants. A suitable loading protocol was optimized by loading the cells at pH 5.6 with 2 μM Fluo-3 for 30 min at 30  °C. Under these conditions, several key factors related to in vitro fertilization were also investigated in order to test their possible effects on the [Ca²⁺]_cyt of the female sex cells. The results indicated that the bovine serum albumin-fusion system was superior to the polyethlene glycol-fusion system for detecting calcium fluctuations in female sex cells during fertilization. The central cell was fertilized with the sperm cell in bovine serum albumin; however, no evident calcium dynamic was detected, implying that a transient calcium rise might be a specific signal for egg cell fertilization.     

Effects of cortisol on immunity in red drum, Sciaenops ocellatus

The immunocompetence of juvenile laboratory-reared red drum treated with cortisol in the diet (250 and 500 mg kg⁻¹ food) was assessed following intramuscular administration of bacterio-phage ( Escherichia coli OX 174), washed bovine erythrocytes (1% suspension) and bovine serum albumin (100mg). Cortisol induced suppressive effects on the fish response to two of the three antigens (i.e., haemagglutinating activity to bovine erythrocytes and precipitin development to bovine serum albumin). These differential effects of cortisol on T-dependent antigens could be interpreted as evidence for heterogeneity of red drum T lymphocytes.     

An additional serum albumin type in the Romanov breed of sheep

29 imported rams and 26 imported ewes (2 to 4 years old), belonging to the native Russian Romanov breed, and their offspring from two subsequent lambings in 1973 and 1974 were tested for serum albumin types.
Four albumin types (D, F, S and W) have been found, with frequencies of 0.18, 0.16, 0.55 and 0.11, respectively.
AID, being the fastest moving albumin type known, has never been found before, and therefore it is considered as a 'new' type.
Available family data support the hypothesis that the synthesis of AID is controlled by a codominant and autosomal allele: AI ^D.     

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.     

Co-ordination between localized wound-induced Ca2+ signals and pre-wound serum signals is required for proliferation after mechanical injury

Abstract. The signals which initiate proliferation of endothelial cells after injury are important for selective blood vessel growth during wound healing or tumour growth. Upon mechanically wounding quiescent cells, a transient [Ca²⁺]_; increase was induced in cells at the wound edge. These same cells proliferated 18-24 h post wounding, as measured by bromodeoxyuridine incorporation. The localized Ca²⁺ signal was required specifically during wounding since blocking Ca²⁺ influx reduced proliferation by 40-50%. Proliferation also required serum since starvation reduced proliferation by 80%. Serum-starved cells proliferated if briefly primed with serum prior to wounding. The signals derived from serum and [Ca²⁺]j combined at least additively to induce proliferation. Therefore, serum priming followed by a single, transient Ca²⁺ signal induced by mechanical injury must occur in a temporally and spatially regulated manner for normal proliferation. Co-ordination between signalling cascades induced by growth factors and release from contact inhibition might be obligatory for localized re-endothelialization after injury.     

Rapid Brain Uptake of Manganese(II) Across the Blood-Brain Barrier

Abstract: ⁵⁴Mn²⁺ uptake into brain and choroid plexus from the circulation was studied using the in situ rat brain perfusion technique. Initial uptake from blood was linear with time (30 s to 6 min) and extrapolated to zero with an average transfer coefficient of ∼6 × 10^-5 ml/s/g for brain and ∼7 × 10^-3 ml/s/g for choroid plexus. Influx from physiologic saline was three- to fourfold more rapid and exceeded that predicted for passive diffusion by more than one order of magnitude. The lower uptake rate from blood could be explained by plasma protein binding as the free fraction of ⁵⁴Mn²⁺ in rat plasma was ≤30%. Purified albumin, transferrin, and α₂-macroglobulin were each found to bind ⁵⁴Mn²⁺ significantly and to restrict brain ⁵⁴Mn²⁺ influx. The results demonstrate that ⁵⁴Mn²⁺ is readily taken up into the CNS, most likely as the free ion, and that transport is critically affected by plasma protein binding. The results support the hypothesis that Mn²⁺ transport across the blood-brain barrier is facilitated by either an active or a passive mechanism.     

Francisella tularensis resistance to bactericidal action of normal human serum

Abstract Lipopolysaccharide and outer membranes from the three virulent encapsulated (Cap⁺) strains of three subspecies of Francisella tularensis and their isogenic avirulent capsule-deficient (Cap⁻) mutants were isolated. It was shown that the Cap⁻ cells and their outer membranes almost completely consumed the available complement of normal human serum whereas Cap⁻ LPS (R-LPS), Cap⁺ cells and their components activated the complement less effectively. Absorption of normal human serum with Cap⁻ strain dramatically reduced the complement consumption for homologous strain and its surface structures. This reduction reflected the loss of bactericidal antibodies. Addition of antibodies to whole cells of F. tularensis completely restored complement activity. The cross-absorbing experiments demonstrated that Cap⁻ cells more effectively deplete bactericidal antibodies than homologous virulent strain. From these results it can be concluded that normal human serum is bactericidal for serum-sensitive Cap⁻ F. tularensis strains through the action of complement initiated by the classical complement pathway and serum resistance of virulent strains is not due to absence of targets for bactericidal antibodies, but is due to their low accessibility because of O-side chains of lipopolysaccharide.     

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.     

Purification and characterization of M3 protein expressed on the surface of group A streptococcal type 3 strain C203

Abstract Monoclonal antibodies (mAbs) have been produced by immunizing BALB/C mice with whole M⁺ bacteria in incomplete Freund adjuvant and the resulting mAbs for M3 protein have been selected by an indirect immuno-fluorescent technique using formaldehyde-fixed M⁺ and M⁻ bacteria. Four mAbs reacted with a 65 kDa protein in an extract obtained from the cell wall of M⁺ bacteria after treatment with N -acetyl muramidase and lysozyme. The purified 65 kDa protein neutralized the phagocytic activity of rabbit anti-M3 antibody. The N-terminal amino acid sequence of the 65 kDa protein was identical with that of protein generated by the M3 gene which has been previously cloned and sequenced. The evidence indicates that the 65 kDa protein is M3 protein. The M3 protein bound not only human fibrinogen but also human serum albumin (HSA). When the M3 protein was purified by gel-filtration and ion-exchange chromatography in the absence of phenylmethyl sulfonyl fluoride (PMSF), four fragments (35 kDa, 32 kDa, 30 kDa, and 25 kDa) in addition to the intact molecule appeared. N-terminal amino acid sequence analysis showed that 35 kDa and 25 kDa fragments were ANAAD and DARSV, respectively, being identical at positions 1–5 and 198–202 to the M3 gene derived protein. Therefore, the 35 kDa and 25 kDa fragments, which were presumed to be cleavage products, may be derived from the C-terminal part and N-terminal part of the intact molecule, respectively. When the effect of purified M3 protein in the bactericidal activity of normal human blood in the presence of M⁻ bacteria was investigated, the M3 protein was responsible for the organism's resistance to attack by phagocytic cells.