Cholesteremia in Japanese quail: response to a mixture of vitamins C and E and choline chloride.

Five-week old, male, Japanese quail (Coturnix coturnix japonica) were given ad libitum access to glucose- soybean meal-10% fat diets containing 0, 0.25, 0.5, or 1% cholesterol, with or without the addition of a vitamin supplement (vitamin C--1 g/kg of diet, vitamin E--30 I.U./kg of diet and choline chloride--5.5 g/kg of diet). After 12 weeks, 9 quail from the 24 quail fed each diet were killed and the total cholesterol concentration of serum, liver, kidney, and aorta was determined. Cholesterol concentrations of these organs increased with increasing levels of dietary cholesterol. The vitamin supplementation enhanced the increase in the cholesterol concentration of serum and kidney, lessened the elevation of the liver cholesterol concentration and had no effect on the aorta cholesterol concentration. The remaining quail were fed the same diets, for a subsequent 12 week period, except that cholesterol was deleted. At the termination of the experiment, the total cholesterol concentration of serum, liver, and kidney returned to control level for all treatments in which organ cholesterol concentrations had been increased previously. Aortic cholesterol concentration decreased during the second 12 week period (0.5 and 1% cholesterol diets fed for the first 12 weeks), however, the aortic cholesterol concentration remained higher than those of the control at 24 weeks. No significant effect of vitamin supplementation on organ cholesterol concentration was noted at 24 weeks although serum cholesterol concentration was significantly lower for the vitamin- fed groups at all levels of dietary cholesterol. Aortic ahteromata were observed at both 12 and 24 weeks in all groups fed 0.5 and 1% cholesterol.


The potential of some synthetic pyrethroids for control of the cattle tick (Boophilus microplus).

As part of a continuing programme to aid in the development of new acaricides, the potentials of three synthetic pyrethroids, permethrin, cypermethrin and decamethrin, were assessed against a range of resistant strains of the cattle tick (Boophilus microplus). The compounds were shown to be acaricidal, with decamethrin the most effective, giving efficient control of susceptible and organo-phosphorus resistant strains of the cattle tick at concentrations as low as one fifth of those previously used for any commercial acaricide. However higher concentrations were required for satisfactory control of the DDT-resistant strain. Permethrin-selection of a field strain containing a low percentage of DDT-resistant ticks led to rapid development of resistance to a level equivalent to that of the DDT-resistant strain. Further selection increased resistance to permethrin to a higher level than that shown in the DDT-resistant strain. This increased resistance to permethrin appears to be restricted mainly to this compound and is not associated with a marked increase in resistance to cypermethrin, decamethrin or DDT. The potential of organo-phosphorus compounds to potentiate the pyrethroids when the two classes of compounds are used as mixtures against the pyrethroid-resistant and susceptible strains was confirmed.


The effect of nutritional restriction of Hereford heifers in late pregnancy.

Two experiments were conducted to examine the consequences of attempting to control dystocia in Hereford heifers by restricting nutrition during late pregnancy. Nutritional level was controlled by varying feedlot rations in experiment 1 and by varying pasture availability in experiment 2. In neither experiment was the incidence of dystocia or stillbirths significantly affected by level of nutrition during late pregnancy. Calf birthweight was reduced and dam pelvic growth was retarded when the feed intake of heifers was restricted prior to calving. Clear evidence of the dangers associated with allowing heifers to lose too much weight in late pregnancy was provided by these experiments. The loss of around 0.5 kg/day during the last trimester of pregnancy was associated with weak labour, increase dystocia rates, increased perinatal mortality, reduced calf growth rate, prolonged post partum anoestrus and a reduced pregnancy rate (not all statistically significant differences). These effects were not evident in heifers which were maintaining or gaining weight at a moderate rate during late pregnancy. On the basis of this work it is suggested that heifers should be fed to allow modest rates of bodyweight gain (0--0.5 kg/day) during late pregnancy.


Toxoplasmosis as a cause of perinatal death in goats.

Lesions typical of congenital toxoplasmosis were found in 5 aborted and stillborn kids. Serological findings in 2 of these kids and all their dams supported the diagnosis of congenital toxoplasmosis. In additon. Toxoplasma gondii was isolated from twin kids.


Vaccination against experimental staphylococcosis in sheep -- observations on bacteriology and pathology following challenge.

Sheep were vaccinated with a killed Staphylococcus aureus vaccine (2 doses) which had been cultured in vitro (Group 1), a killed S. aureus vaccine (2 doses) cultured in vivo (Group 2) or a single dose of a live vaccine (Group 3). Other sheep were used as non-vaccinated controls. All sheep were challenged by intravenous injection of 2.6 x 10(11) washed, viable S. aureus organisms, the vaccinated animals being given the challenge inoculum at various intervals after vaccination. The control sheep survived for 29h (mean) after challenge. Animals given killed vaccines survived longer, (particularly Group 2) if challenged less than 40 days post-vaccination, compared with those challenged more than 40 days post-vaccination. Animals in Group 3 survived longer if challenged after 40 days post-vaccination than those in Groups 1 or 2. There were no significant differences between the treatment groups for numbers of S. aureus recovered from blood in the 3h period following challenge. Histological and bacteriological evidence showed that the kidneys were more severely affected by the challenge inoculum than heart, spleen, liver or lungs. The kidneys showed both toxigenic and lymphoreticular reactions and large numbers of staphylococci were recovered more reliably from kidneys than other organs.


Copper administration to young calves: oral dosing with copper oxide compared with subcutaneous copper glycinate injection.

Calves were dosed with 50 g of copper oxide granules orally or given 120 mg copper glycinate subcutaneously. There was no significant difference between the weights of calves given either treatment during the trial. However, the copper oxide treatment resulted in sustained higher concentration of plasma copper and higher liver reserves than the glycinate treatment.


Atypical porcine enterovirus encephalomyelitis: possible interraction between enteroviruses and arsenicals.

Porcine enteroviruses were isolated from weaner pigs that had nervous signs and mild non-suppurative meningoencephalomyelitis and ganglioneuritis. The clinical signs and lesions were not typical of enterovirus infection and it is believed that an organic arsenical present in feed enhanced pathogenicity of enteroviruses. Severe non-suppurative polioencephalomyelitis and ganglioneuritis were produced in gnotobiotic pigs by oral inoculation of the viruses.


Ossifying ameloblastoma in a horse.

The features of an ossifying ameloblastoma in a 5-year-old gelding are described. The tumour developed in the angle of the right mandible and microscopically consisted of multiple follicles and islands of epithelial tissue adjacent to which were trabeculae of bone, osteoid and compact collagenous tissue.


The role of inhibited larvae in the epidemiology of ovine haemonchosis.

Three paddocks were contaminated with Haemonchus contortus eggs from early spring to mid summer by yearling sheep. In Paddock 1, all contamination was derived from worms that had been previously inhibited in their development. The contamination in Paddock 2 was produced only by infections recently acquired from pasture, and in Paddock 3 contamination was attributable to both sources. When the yearling sheep were replaced in mid-summer by newly weaned lambs, severe outbreaks of haemonchosis occurred on all paddocks. It was concluded that inhibited H. contortus larvae are capable of producing sufficient eggs, when they resume development, to initiate an outbreak of haemonchosis in susceptible sheep.


A comparison of urinary specific gravity and osmolality in sheep.

Urinary specific gravity and osmolality were determined on urine samples collected from three categories of sheep -- animals that were normal, in diuresis or had nephron numbers reduced surgically. Values for specific gravity and osmolality were compared and regression coefficients calculated for each category of sheep. In this study, specific gravity was found to be a relatively reliable indicator of osmolality, the correlation being highest in the urine from the sheep with reduced nephron numbers.


A retrospective study of osteomyelitis in dogs and cats.

Thirty-nine cases of osteomyelitis in dogs and cats were recorded at the Sydney University Veterinary Hospital and Clinic over a three and a half year period. In 36 cases osteomyelitis was established prior to admission. Three cases of osteomyelitis became established from a total of 502 orthopaedic surgery cases seen at the hospital in this period. In the dog the most common source of infection was open reduction of closed fractures, while in the cat, the most common source of infection was an extension from soft tissue infection. More males than females were affected. Ten cases of osteomyelitis were treated successfully, twelve cases required amputation, while euthanasia was performed on thirteen other cases. The problems and principles of treatment of active osteomyelitis as reflected in the treatment of this series of cases have been described.


Benzimidazole resistance in small strongyles of horses.

Treatments with mebendazole, cambendazole or febantel were ineffective in reducing the faecal egg count of a group of 40 horses infected with small strongyle species. Evidence was also obtained that this apparent resistance extended to fenbendazole. In horses dosed with morantel tartrate egg counts were reduced to zero by 7 days after treatment. The development of resistance is discussed in relation to previous treatment of the horses and the mode of action of febantel.


Effect of some pesticides on micro-organisms in the rhizosphere of maize, common bean, or cotton.

Pot experiments were run to investigate the influence of temik (insecticide and nematocide) and of orthocide (fungicide) on the soil micro-organisms with maize, common bean, and cotton. The counts showed that in some cases there was no influence on the micro-organisms compared with the control, in some cases their numbers increased. On the other hand, depressive effects were recorded for varying periods. This supports the evidence that the pesticides are attacked by several types and groups of rhizospheric microflora; in some cases the by-products may be more toxic than the original toxicants. Generally, however, the recorded effects differed according to the chemical composition of the pesticides, the type and growth phase of each individual plant, and the micro-organisms concerned.


Interaction of alpha-thalassemia genes with each other and with HbC in an American black family.

The relative rates of in vitro synthesis of hemoglobin chains have been studied in an American black family in which the mother is doubly heterozygous for alpha-thalassemia and HbC and the father is heterozygous for alpha-thalassemia. The alpha/non-alpha synthetic ratio was equally unbalanced in both the bone marrow and the peripheral blood of the mother. Although HbC comprised 35% of her hemoglobin (compared to 42.2 +/- 2.2 in individuals with HbC trait and balanced globin synthesis), synthetic data showed that the newly synthesized beta C chain was 44% of the total newly synthesized beta chains. Isolated membranes contained more newly synthesized beta C than beta A chains. Three of the offspring were within the normal range, and the remaining three had alpha-thalassemia. There were two spontaneous abortions during the second trimester of pregnancy. Hydrops fetalis did not occur, and none of the children had HbH disease or HbC trait.


Human salivary proline-rich (Pr) proteins: a posttranslational derivation of the phenotypes.

The acidic proline-rich proteins (Pr) showing genetic polymorphism were purified from human parotid salivas by gel filtration and ion exchange chromatography. Molecular weight determinations, amino acid composition analyses, and polypeptide mapping experiments indicate that the Pr 3 protein is a fragment of the Pr 1 protein. Studies of a parotid saliva factor capable of converting Pr 1 to Pr 3 and Pr 2 to Pr 4 indicate that Pr 3 and Pr 4 are generated from Pr 1 and Pr 2, respectively. Evidence suggests that the converting factor is a protease capable of posttranslationally cleaving Pr 1 and Pr 2, the primary or derived products of alleles Pr1 and Pr2.


Genetic basis of creatine kinase isozymes in skeletal muscle of salmonid fishes.

The genetic basis of isozyme phenotypes of creatine kinase (CK) from extracts of skeletal muscle of salmonids has been resolved through breeding data including double heterozygous crosses and backcrosses of rainbow trout (Salmo gairdneri), and backcrosses of coho salmon (Oncorhynchus kisutch). The two-three-, or four-banded phenotypes of homozygous individuals and all heterozygous and hybrid phenotypes of ten salmonid species are readily explained by the following model: (1) there are no detectable heterodimers either between allelic products at a single locus or between loci: (2) each allele is represented electrophoretically by two bands, presumably a reflection of stable posttranslational modification of a single polypeptide unit; (3) CK of salmonid muscle is encoded by two loci--CK-1 and CK-2. The distance separating the paired bands reflecting each allele provides a basis for two groupings--a broad-spaced group (including all species of Oncorhynchus tested excepting O. masou) and a narrow-spaced group (including all species of Salmo tested and O. masou). The relationships among species suggested by the relative mobilities and spacings of these CK bands are consistent with taxonomic schemes inferred from morphological, cytogenetic, and other isozymic data.


Genetic variation in alkaline phosphatase of the house mouse (Mus musculus) with emphasis on a manganese-requiring isozyme.

Genetic variation among inbred strains is described for electrophoretic migration of alkaline phosphatase from intestine, kidney, blood plasma, and three isozymes of liver. A manganese-requiring isozyme of liver and kidney unaffected by neuraminidase is described, and the locus controlling variation in this isozyme is designated Akp-1. Data from recombinant inbred strains place the locus on chromosome 1 at a distance of 3.6 +/- 2.9 cM from the M1s locus on the side distal to the centromere. Test-cross data show the following gene order and recombination percentages: Dip-1 19.0 +/- 3.8% Lp 7.4 +/- 2.2% Akp-1.


Tissue-specific zein synthesis in maize kernel.

Zein may account for as much as 10% of the total protein in the mature embryo of maize inbred W64A. This protein exhibited an electrophoretic pattern on SDS gels similar to that of the endosperm. Like the endosperm system, the synthesis of zein components in the embryo was controlled by the opaque-2 and floury-2-mutations. However, unlike zein synthesis in the endosperm, zein synthesis in the embryo could not be increased by nitrogen fertilizer. Variations in amino acid composition were observed between the zein components of the embryo and those of the endosperm.


Serum esterase genetics: identification and hormone induction of the Es-1b esterase in inbred rats.

A previously unrecognized esterase from the sera of the appropriate strains of the rat Rattus norvegicus was revealed by a discontinuous polyacrylamide gel electrophoretic technique. This esterase migrated in the albumin region, whereas a previously known major albumin esterase controlled by the Es-2 locus migrated in the postalbumin region when the method was used. The new albumin esterase component which separated from the Es-2 esterase was identified as the product of the Es-1b gene. The new albumin esterase was not detectable in the sera of sexually mature males of the appropriate genotype, because the activity level of this esterase was influenced by sex hormones, especially androgen.


An electrophoretic method for the detection of human red cell acetylcholinesterase.

An electrophoretic method to demonstrate human red cell membrane acetylcholinesterase is described. The method, performed with crude Triton lysates of red cells on a flat-bed polyacrylamide gel, is sufficiently simple to be suitable for a population survey.


Amino acid sequence differences in pancreatic ribonucleases from water buffalo breeds from Indonesia and Italy.

The amino acid sequences of the pancreatic ribonucleases from river-breed water buffaloes from Italy and swamp-breed water buffaloes from Indonesia differ at three positions. One of the differences involves a replacement of asparagine-34, with covalently attached carbohydrate on all molecules, in the river-breed enzyme by serine in the swamp-breed enzyme. The ribonuclease content of the pancreas differs considerably between breeds and is lower in river buffaloes. A ribonuclease preparation from two swamp buffaloes contained a minor glycosylated component. Preliminary evidence was obtained that the amino acid sequence of this component has factors in common with the main component of the swamp-breed ribonuclease and with the river-breed enzyme.


Floride inhibition in patients with atypical dibucaine-resistant plasmacholinesterase.

The fluoride numbers (FN) of 144 individuals referred to our laboratory for dibucaine number (DN) determinations because of prolonged apnea following succinylcholine in the probands or relatives were determined by our new method using 2.5 X 10(4) M concentration of sodium fluoride at 37 C. Dibucaine-resistant homozygotes and heterozygotes both had lower FN than the normal homozygotes. A linear correlation between DN and FN for all these genotypes was found which is described by the equation FN = 0.59 DN + 32. The correlation coefficient was r = 0.94. This equation will help in the correct identification of atypical fluoride resistant genotypes, since these ought to have an observed FN lower by 2 standard deviations than the FN calculated from this equation.


Identification of a new allele of Es-I segregating in an inbred strain of mice.

A new allele of Es-1, designated Es-1e, has been identified in the mouse.  This allele was discovered segregating among the progeny of a strain DBA/2J male and is apparently the result of a spontaneous mutation within this strain. Genetic analyses have shown that this mutation is heritable and, further, that both heterozygous and homozygous progeny are viable and fertile. To date, no discernible deleterious effects have been identified as associated with this mutation.


Evidence for a triplicate set of glucosephosphate isomerase structural genes in hexaploid wheat.

The glucosephosphate isomerase (GPI)zymogram phenotypes of 46 aneuploid derivatives of the cultivar Chinese Spring of hexaploid wheat were determined. Variation was observed among the strains in the relative level of expression of three GPI isozymes.  The relationships observed between chromosomal constitution and zymogram phenotype support the hypothesis that the three GPI isozymes are dimers composed of protomers encoded by a minimum of three homoeologous structural genes located one each in the short arms of chromosomes 1A, 1B, and 1D.  The relative levels of expression per dose of chromosome arm of the products of the three arms differ in a manner consistent with the presence of a two-fold greater quantity of the product of 1BS than of the product of 1AS and of 1DS, indicating that 1BS may contain duplicate GPI structural genes.


Genetic variation, inheritance, and quaternary structure of malic enzyme in brook trout (Salvelinus fontinalis).

Electrophoretic variation is described for malic enzyme (ME) for the first time in brook trout (Salvelinus fontinalis).  Since the quaternary structure of ME was not clear from examination of banding patterns in brook trout alone, ME phenotypes in rainbow trout (Salmo gairdneri)X brook trout hybrids as wel as in esocid species demonstrated that ME is tetrameric.  A model of two duplicated loci is proposed to account for the observed variation.  One locus (ME-2) is fixed and one locus (ME-1) is variable with three electrophoretically distinct alleles; the protein products of ME-1 are reduced in activity relative to the protein products of ME-2.  Joint segregation was examined between ME-1, ME-2, and nine other biochemical loci in a splake--lake trout (Salvelinus namaycush) X brook trout hybrid--backcross.  All pairwise examinations showed random assortment except ME-2 with an isocitrate dehydrogenase locus (IDH-3), which showed complete linkage in the splake backcross.  This may be due to a chromosomal aberration.


Genetic variation in Hawaiian Drosophila. VIII. Heterozygosity and genic changes in isolated populations of D. engyochracea.

Drosophila engyochracea, an endemic Hawaiian fly found only in two, finite populations in Volcano National Park, has extensive electrophoretic heterozygosity on a par with that found in species with much wider distributions. A study of six polymorphic loci in both populations over an 18-month period revealed that the population in the more xeric environment is more dynamic genetically as well as more variable. In addition, genetic changes at one locus, Pgm, are correlaed to changes in an environmental moisture parameter. These findings confirm that migration is not necessary to maintain genetic variation in isolated population and demonstrate that D. engyochracea gene pools are susceptible to errors in Hardy-Weinberg equilibria during specific seasonal periods.


Mitochondrial malic enzyme in mosaic skeletal muscle of mouse chimeras.

The question was investigated whether mitochondria in the mammalian skeletal muscle fiber syncytium incorporate gene products encoded by one or many nuclei.  Mouse chimeras were produced from strains which differ in their electrophoretic variants of the nuclear-coded mitochondrial protein, malic enzyme (MOD-2, E.C. 1.1.1.40, L-malate NADP+ oxidoreductase decarboxylating).  The MOD-2 phenotypes of skeletal muscles of these chimeras were characterized in a starch gel electrophoretic system.  The results indicate that individual mitochondria can contain products encoded by multiple nuclei and therefore that, for skeletal muscle mitochondria, the cell is not subdivided into nuclear territories.  Possible mechanisms of gene product distribution in skeletal muscle fibers are discussed.


Biochemical characterization of mutant propionyl CoA carboxylases from two minor genetic complementation groups.

We have biochemically characterized several parameters of propionyl CoA carboxylase (PCC) activity in fibroblast extracts from PCC-deficient patients belonging to the two minor genetic complementation groups with those of the two major complementation pccB and pccBC. Comparison of PCC's from these groups with those of the two major complementation groups, pcc A and pcc C, has demonstrated that PCCs from both the pcc B and pcc BC groups closely resemble each other as well as PCC from the pcc C group.  These results further support the hypothesis that the pcc B and pcc BC lines are interallelic with respect to pcc C and consequently that the structural mutations in the PCCs from these groups involve the same subunit.


Evaluation of biotin responsiveness in cultured fibroblasts from patients with propionic acidemia: absence of response by structurally altered carboxylases.

We have demonstrated that, although propionyl CoA carboxylase (PCC) activity is deficient in fibroblast extracts from PCC-deficient patients belonging to the two major and two minor genetic complementation groups, the activity of another biotin-dependent carboxylase, beta-methylcrotonyl CoA carboxylase (betaMCC), is normal.  Moreover, betaMCC activity is stimulated when the fibroblasts are cultured in high concentrations of biotin, in the same way that it is in normal fibroblasts, whereas the depressed PCC activity remains essentially unchanged.  Because these results are parallel with the in vivo failure of high-dose biotin to stimulate PCC activity in peripheral blood leukocytes, we conclude that the biotin responsiveness of PCC in cultured fibroblasts from patients with PCC deficiency may be  used to predict or confirm biotin responsiveness in vivo.


Cadmium metabolism in cdm/cdm mice.

The uptake and metabolism of cadmium by cadmium-susceptible (+/+) and cadmium-resistant (cdm/cdm) strains of mice have been compared. These strains did not differ with respect to the quantitative uptake of cadmium into liver, kidney, or testis.  After intraperitoneal administration of a nontoxic dose, more than 80% of the cadmium in liver and testis of both strains is bound to low molecular weight proteins.  The chromatographic behavior of these cadmium-binding proteins is not affected by cdm genotype.


Structural relatedness of mouse lactate dehydrogenase isozymes, A4 (muscle), B4 (heart), and C4 (testis).

Three homotetrameric lactate dehydrogenase isozymes, LDH-M(A4), LDH-H(B4), and LDH-X(C4), from DBA/2J mice have been purified by affinity chromatography.  The amino acid compositions of the subunits A,B, and C, based on a molecular weight of 36,000, have been determined.  The compositional relatedness of these isozymes indicates that subunits A (muscle) and B (heart) are more closely related to each other than to subunit C (testis).  Tryptic peptide maps and amino acid compositions of some active site peptides apear to confirm the compositional relatedness among these isozymes.  The sequence of the loop region of mouse C subunit seems to be markedly different from all known A and B sequences, and the structural and functional implications are discussed.


Developmental expression of murine HPRT. I. Activities, heat stabilities, and electrophoretic mobilities in adult tissues.

Total and specific activity of the enzyme hypoxanthine phosphoribosyltransferase (HPRT) varied widely among six tissues from C3H/f mice; the highest levels of activity were in brain. More striking were thermostability differences in tissue enzymes. Although brain, spleen, and kidney HPRT retained 65% basal activity after 15 min at 85 C, heart, liver, and erythrocyte HPRT retained only 20-30% initial activity. Kidney HPRT behaved as monospecific heat-stable enzyme (K-denatauration=0.022/min, and liver enzyme behaved as monospecific heat-labile enzyme (K-denaturation=0.061/min), while other tissues appeared to contain both forms of the enzyme. Multiple electrophoretic activity bands were present in all tissues; no activity band was restricted to a single tissue. The data presented here are consistent with the hypothesis that the distinct tissue properties of HPRT result from posttranslational modification of the product of a single genetic locus which is expressed in all tissues.


Genetics of cat hemoglobins: a quantitative polymorphism.

Blood samples from domestic cats, representing the parents (26) and offspring (91) from 33 matings, were analyzed to determine the proportions of hemoglobins A and B. Evidence is presented that the proportions of the two hemoglobins are genetically determined and that cats may be divided into three groups on the basis of this characteristic. Family data and Hardy-Weinberg analysis support the hypothesis that the three groups represent the possible combinations of an allelic gene pair. In an attempt to explain the observed phenotypic differences, possible points of action of this gene pair are discussed, including effects on rates of synthesis or epigenetic modification of globin chains.


Evidence for 15 genetically determined electrophoretic variants of transcobalamin II in rabbit serum.

By starch gel electrophoresis and autoradiography two classes of vitamin B12 binding proteins were detected in rabbit serum. By analogy to the nomenclature used in man, the two classes of proteins were named "transcobalamin I" (TCI) and "transcobalamin II" (TCII). Fifteen TCII phenotypes were observed, and family data indicated  that they are controlled by five allelic codominant genes. The possibility that the five genes arise from the action of at least two polymorphic and closely linked structural loci is discussed.


Sex difference in the accumulation of D-beta-aminoisobutyrate in organs of mouse after thymine loading.

The concentration of D-beta-aminoisobutyric acid (D-BAIB) in the liver and kidney was twice as high and dropped more slowly in the female mouse than in the male after an intraperitoneal injection of thymine. The concentration of beta-alanine, formed from uracil by the same enzyme system catalyzing formation of D-BAIB from thymine, was not different in the liver and kidney of both sexes after an intraperitoneal injection of uracil. After the intraperitoneal injection of D-BAIB, the concentration of BAIB in male liver decreased faster than that in female liver. Inhibition of D-BAIB: pyruvate aminotransferase caused by injection of D-cycloserine resulted in a significant increase in the concentration of BAIB in liver of both sexes after injection of thymine, but the concentration dropped more rapidly in the male. The activity of D-BAIB: pyruvate aminotransferase was not different in the livers of male and female mice. Under the action of probenecid, an inhibitor of active transport systems, the sex difference in accumulation and disappearance of the amino acid in the liver was not observed. This suggested that the excretion of BAIB is more active in the renal tubules of the male mouse than in those of the female. However, the amount of BAIB excreted in the urine after injection of thymine was larger in the female mice than in the male mice. There may be another probenecid-sensitive enzyme for the disposal of BAIB in male mice.


6-phosphogluconate dehydrogenase in the housefly, Musca domestica L.:evidence for inheritable 6PGD polymorphism.

Two electrophoretic variants of the 6-phosphogluconate dehydrogenase (6 PGD) enzyme have been found in the WHO/IN/Musca domestica/l housefly laboratory strain. The patterns shown by Cellogel zone electrophoresis can be fully explained by the hypothesis of two codominant autosomal alleles. On this hypothesis, a specific Pgd locus has been postulated and the symbols PgdA and PgdB have been assigned to the two alleles causing the PGD-A and PGD-B phenotypes. The bands corresponding to the homozygous phenotypes PGD-A and PGD-B have different electrophoretic mobility and staining intensity; they can be described, respectively, as "fast-weak" and "slow-thick." The heterozygous phenotype PGD-AB gives a three-banded pattern, indicative of a dimeric structure for this enzyme; this pattern is asymmetrical. Heterozygous flies have been found both among wild-type strains of recent colonization and among old established laboratory colonies. Most strains are PgdB monomorphic; up to now only three strains have been PgdA monomorphic, all of them being multimarker strains. The Pgd locus has been traced to the housefly linkage group III.


Aryl hydrocarbon hydroxylase activity in pulmonary alveolar macrophages and lymphocytes from lung cancer and noncancer patients: a correlation with family histories of cancer.

Aryl hydrocarbon hydroxylase (AHH) activity was measured in pulmonary alveolar macrophages (PAMs) and peripheral blood lymphocytes from cigarette smokers with and without primary lung cancer. Frequency distribution analysis of AHH induction ratios for the two groups revealed an increased number of individuals in the lung cancer patient group with high lymphocyte induction values (P less than 0.05). A similar increase was not shown for high-PAM AHH values in lung cancer patients (P greater than 0.2). When individual PAM and lymphocyte AHH values were compared between noncancer and lung cancer patients, a positive correlation was observed for noncancer patients (r=0.195, P less than 0.001), but no correlation of these values was noted for lung cancer patients. The lung cancer patients were divided into three subgroups of patients showing (I) high PAM and low lymphocyte AHH levels, (II) low PAM and low lymphocyte AHH levels, and (III) low PAM and high lymphocyte AHH levels. When the incidence of family history of cancer was compared for these subgroups, no family cancer history was recorded for persons in subgroup II; however, individuals in subgroups I and III presented family cancer history incidence of 9.5% and 39.3%, respectively. Patients in group III averaged 6 years younger than those in group I. These data suggest that familial factors may be identified among lung cancer patients and that these factors appear to associate as either a cause of an effect with the capacity of pulmonary alveolar macrophages and lymphocytes to be induced for AHH. The data support the hypothesis that high AHH values may be characteristic of lung cancer patients but show that enzyme values determined from a single tissue, either PAMs or lymphocytes, may not be appropriate for showing whether high AHH inducibility is correlated with lung cancer.


Genetic polymorphism of vitamin B12 binding (R) proteins of human saliva detected by isoelectric focusing.

Genetic polymorphism of the vitamin B12 binding (R) proteins of parotid saliva is determined by autosomal inheritance of codominant alleles. This hypothesis is supported by studies in 43 families including 152 children. For randomly collected salivas from 143 whites, 104 blacks, and 75 Chinese, gene frequencies are as follows: for whites, Rs1=0.88, Rs2=0.12; for blacks, Rs1=0.94, Rs2=0.06; for Chinese, Rs1=1.00. This genetic marker is also shared by R proteins of milk, tears, and leukocytes. In the Rs 1--2 salivary type there is less labeling of the protein products of Rs2 v. Rs1 with 57Co B12 as assessed by the intensity of the bands on the autoradiogram. There is no evidence for close linkage (theta less than 0.01) between Rs and TC II (transcobalamin II) or between Rs and salivary protein locus Pr, Db, Gl, or Ps.


Genetics of the mammalian phenylalanine hydroxylase system. II. Immunological and two-dimensional gel electrophoretic studies of phenylalanine hydroxylase in cultured normal and mutant rat hepatoma cells.

We have examined 11 previously described cultured rat hepatoma mutants with absent or reduced phenylalanine hydroxylase activity (Choo and Cotton, 1977). Immunological and electrophoretic methods failed to detect any structurally altered protein in these mutants. In nine independently isolated revertants from four different mutants, wild-type protein was regained (or accentuated). This evidence suggests that the mutation involved in these mutants is most likely to be regulatory in nature. These studies have provided three reasons for believing that in cultured rat hepatoma cells one gene codes for a single polypeptide chain, a number of which combine to form the active phenylalanine hydroxylase multimer: (1) Analysis of the purified protein by two-dimensional electrophoresis revealed only a single polypeptide chain. (2) This polypeptide was diminished or undetectable in crude extracts of 11 independently isolated mutants with absent of reduced activity. (3) In none of these 11 mutants was the polypeptide we have designated to be phenylalanine hydroxylase present at normal levels, as would be expected if the mutation were at another locus responsible for a possible second subunit.


Isoenzyme pattern of HPRT in murine erythrocytes: control by an autosomal locus.

A locus on chromosome 7 controls the electrophoretic mobility of hypoxanthine phosphoribosyltransferase (HPRT) isoenzymes in mouse erythrocytes, but not in several other tissues. This locus is designated Hma (HPRT mobility alteration) and maps very close to the Hbb locus. The A/J, AKR/J, AU/SsJ, BALB/cJ, CBA/J, C3H/HeJ, DBA/2J, LP/J, RF/J, SEA/Gn, ST/BJ, and 129/J strains and our population of Swiss albino mice have the Hmaa allele. Hmaa is dominant to Hmab, which is found in the C57BL/6J, C57BL/KsJ, C58/J, LT/Sv, MA/MyJ, SJL/J, and SWR/J strains. Both alleles are found in feral Mus musculus. In our conditions, homozygotes for Hmab have two major bands of HPRT activity after electrophoresis of extracts of erythrocytes and of other tissues. Heterozygotes and Hmaa homozygotes have three bands in erythrocyte extracts but two band in other tissues.


Intracellular mechanisms of the formation of homo- and heteropolymeric isozymes. I. Kinetics of the formation of the heteropolymeric isozyme of malate dehydrogenase in parasexual hybrids of two Acetabularia species.

The isozyme pattern of malate dehydrogenase (MDH) of Acetabularia crenulata and A. mediterranea is characterized by heterogeneity in different regions of the cytoplasm of both algae, as well as by species specificity. The formation of the heteropolymeric MDH isozyme is restricted to a definite region of the cytoplasm of heterokaryons and nuclear-cytoplasmic A.crenulata-A.mediteranea hybrids at different stages of their development. The data obtained suggest that the concentrations of the free subunits of MDH, coded for by homologous genes, are unevenly distributed in the cytoplasm of hybrid cells. The heteropolymeric MDH isozyme in these cells is presumably the result of the de novo synthesis of isozyme subunits. This seems plausible inasmuch as no exchange occurs between the homopolymeric MDH isozymes of both parental types in the cytoplasm. The formation of the heteropolymeric MDH isozyme is tentatively related to the spatial compartmentalization of the mRNAs of homologous genes coding for the MDH subunits.


Role of 24- and 28-hydroxylated intermediates in the metabolism of beta-sitosterol in the insect Tenebrio molitor.

1. [28-3H]Stigmast-5-ene-3 beta, 28-diol and [23,23,25-3H]stigmast-5-ene-3 beta, 24-diol were synthesized. 2. Each of the samples was mixed with beta-[4-14C]sitosterol and administered to Tenebrio molitor larvae. 3. The former compound is not utilized by the insect; the latter, although metabolized to 24(28)-ethylidene sterols and cholesterol, is not a beta-sitosterol metabolite. 4. The above results are discussed in relation to the mechanism of formation of the 24(28)-double bond in beta-sitosterol metabolism in T. molitor.


The nature of the slow metal ion-dependent conformational transition in bovine prothrombin.

Kinetic parameters characterizing the slow structural isomerization observed via metal ion-dependent intrinsic fluorescence quenching of bovine prothrombin Fragment 1 have been determined. From forward and reverse rate constants, an equilibrium constant of approx. 0.25 is calculated. This result is consistent with the hypothesis that there exists, in the absence of metal ions, an equilibrium between two forms of bovine Fragment 1, one of which can interact rapidly with Ca2+ and subsequently with phospholipid. The other form of Fragment 1 cannot interact with Ca2+ in a manner that yields a phospholipid-binding form of the protein. Interconversion of these two forms of Fragment 1 occurs and may involve the isomerization of a proline residue.


Heparin-sepharose affinity chromatography for purification of bull seminal-plasma hyaluronidase.

Bull seminal-plasma hyaluronidase was purified 180-fold by chromatography on concanvalin A-Sepharose, heparin Sepharose, Sephadex G-200 and Sephacryl S-200. With hyaluronic acid as the substrate, the specific activity and turnover number of purified hyaluronidase were 3.63 mumol/min per mg (104000 National Formulary units/mg of protein) and 214 min-1 (mol of product formed/mol of enzyme per min) respectively. Polyacrylamide-gel electrophoresis indicated that the purified enzyme migrated as a single band on 7.5 and 10% (w/v) gels at pH 4.3 and 5.3. Bull seminal-plasma hyaluronidase was markedly inhibited by hydroxylamine, phenylhydrazine and semicarbazide. Purified hyaluronidase (1.25 munits; 1 unit = 1 mumol of N-acetylglucosamine liberated/min at 37 degrees C) dispersed the cumulus clot of rabbit ova in 1 h at 22 degrees C.


Matrix proteins bound to associatively prepared proteoglycans from bovine cartilage.

Proteoglycans were extracted from bovine tracheal cartilage by high-speed homogenization, the use of dissociative solvents being avoided. The homogenate was fractionated by gel chromatography, sucrose-density-gradient centrifugation and ion-exchange chromatography. A previously unrecognized protein, cartilage matrix protein, was identified by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. It cofractionated with the proteoglycans in all systems, indicating an interaction. The cartilage matrix protein-proteoglycan complex was dissociated by treatment with 4M-guanidinium chloride. The complex again formed when the guanidine was removed. The cartilage matrix protein has a mol.wt. of more than 200000. On reduction it yields subunits with a mol.wt. of approx. 60000.


The isolation and characterization of bovine C4a, an activation fragment of the fourth component of complement.

The fourth component of bovine complement, C4, was cleaved specifically by subcomponent C1s to produce two fragments, C4a and C4b. The smaller, C4a, was isolated in pure form and is a peptide of 9500 mol.wt. containing approx. 84 amino acids and no detectable carbohydrate. C4a has an amino acid composition that is comparable with the anaphylatoxins C3a and C5a, containing six cysteine residues/mol and a high proportion of basic residues. The amino acid sequence of the first thirteen residues shows four identities with the porcine C3a sequence. There is almost complete identity between the C4a sequence and that of the alpha-chain of human C4, indicating that this region is highly conserved. This evidence also clearly establishes that C4a is cleaved from the N-terminal of the alpha-chain of C4.


The purification and characterization of subcomponent C1s of the first component of bovine complement.

Bovine C1s, a subcomponent of the first component of complement, was purified in good yield by a combination of euglobulin precipitation and ion-exchange and molecular-sieve chromatography. Approx. 10 mg can be obtained from 3 litres of serum, representing a yield of 11%. The C1s is obtained in zymogen form, with a mol.wt. of 85000-88000, determined by gel filtration and SDS/polyacrylamide-gel electrophoresis. It is haemolytically active when tested with human C1q and C1r. Activation can be achieved by incubation with human C1r, resulting in cleavage of the C1s chain into two chains of 65000 and 27000 mol.wt. and the generation of an isoleucine N-terminal residue on the smaller chain. Active C1s binds an equimolar amount of di-isopropyl phosphorfluoridate to the smaller chain, which is the C-terminal part in the zymogen. The chains can be separated by ion-exchange in 8 M-urea. All of these characteristics show that bovine C1s is very similar to its human counterpart.


Specificity for guanine nucleotide activation and stabilization of rabbit cardiac adenylate cyclase.

These studies examined the structural specificity for guanine nucleotide-facilitated hormonal activation and guanine nucleotide stabilization of cardiac adenylate cyclase. 1. The phosphonate analogues of GTP, p[CH(2)]ppG (guanosine 5'-[betagamma-methylene]-triphosphate) and pp[CH(2)]pG (guanosine 5'-[alphabeta-methylene]triphosphate), were the most effective activators of adenylate cyclase. Other nucleotides producing significant activation (P<0.01) were, in decreasing order of activation: ITP, GDP, GMP, GTP, XTP, CTP, p[NH]ppG (guanosine 5'-[betagamma-imido]triphosphate), dGTP and 2'-O-methyl-GTP. Guanosine, cyclic GMP, UTP and ppppG (guanosine tetraphosphate) had no effect, and 7-methyl-GTP caused a decrease in the activity. 2. Preincubation of membranes at 37 degrees C for 15min before assay at 24 degrees C produced an 80% decrease in adenylate cyclase activity, and preincubation with p[CH(2)]ppG and pp[CH(2)]pG protected and resulted in a net increase in activity. Other nucleotides that completely or partially preserved activity in decreasing order of effectiveness were p[NH]ppG, GDP, GTP, dGTP, ITP, ppppG, 2'-O-methyl-GTP, GMP, CTP and XTP. Several compounds had no effect, including guanosine, cyclic GMP and UTP, whereas preincubation with 7-methyl-GTP produced a further decrease (P<0.05) in activity. 3. The concentration-dependence for activation and stabilization by the naturally occurring guanine nucleotides was examined in the absence of a regenerating system and revealed GMP to have no stabilizing effect and to be less potent than either GDP or GTP in activating adenylate cyclase. 4. A significant correlation (r=0.90) was found between the properties of activation and stabilization for the compounds examined. These findings are consistent with there being a single nucleotide site through which both the activation and stabilization of adenylate cyclase are mediated.


Nucleotide sequences of chloroplast 5S ribosomal ribonucleic acid in flowering plants.

Evidence for the sequence of duckweed (Lemna minor) chloroplast 5S rRNA was derived from the analysis of partial and complete enzymic digests of the 32P-labelled molecule. The possible sequence of the chloroplast 5S rRNA from three other flowering plants was deduced by complete digestion with T1 ribonuclease and comparison of the sequences of the oligonucleotide products with homologous sequences in the duckweed 5S rRNA. This analysis indicates that the chloroplast 5S rNA species differ appreciably from their cytosol counterparts but bear a strong resemblance to one another and to the 5S rRNA species of prokaryotes. Structural features apparently common to all 5S rRNA molecules are also discussed.


4.5S ribonucleic acid, a novel ribosome component in the chloroplasts of flowering plants.

A species of low-molecular-weight ribosomal RNA, referred to as '4.5S rRNA', was found in addition to 5S rRNA in the large subunit of chloroplast ribosomes of a wide range of flowering plants. It was shown by sequence analysis that several variants of this RNA may occur in a plant. Furthermore, although in most flowering plants the predominant variant contains about 100 nucleotides, in the broad bean it has less than 80. It seems, therefore, to be much more diverse in size and sequence than the other ribosomal RNA species. Like 5S rRNA , it does not contain modified nucleotides and it is also unusual in having an unphosphorylated 5'-end. It is apparently neither a homologue of cytosol 5.8S rRNA nor a fragment of 23S rRNA.


Quantitative N-terminal analysis of fibrinogen-fibrin-related antigen [FR antigen] from human plasma.

Fibrinogen-fibrin-related antigen (FR antigen) was isolated from as little as 1 ml of human plasma by immuno-affinity chromatography with agarose-bound antibody to human fibrinogen. N-terminal analysis was performed to determine the nature and extent of proteolytic degradation of the FR antigen in patients with disseminated intravascular coagulation and in normal subjects. Thrombin cleavage of the A- and B-peptides from fibrinogen in vitro was monitored by the appearance of N-terminal glycine, and an increase in glycine was shown in the FR antigen of patients with disseminated intravascular coagulation. As plasmin progressively degraded fibrinogen, increases in N-terminal alanine, aspartic acid and lysine were observed, corresponding to the known plasmin-cleavage points of fibrinogen; increases in these N-terminal amino acids were also found in the patients' FR antigen. Thrombin treatment in vitro was used to remove fibrinopeptide A (N-terminal alanine) from the samples and to reflect specifically the N-terminal alanine at the plasmin-cleavage point (Arg-42-Ala-43) of the B beta-chain on assay; this alanine was increased progressively in the FR antigen of a patient during urokinase therapy, and was high in other patients when the FR antigen was examined by this procedure.


Does high-mobility-group non-histone protein HMG 1 interact specifically with histone H1 subfractions?

The interaction of the non-histone chromosomal protein HMG (high-mobility group) 1 with histone H1 subfractions was investigated by equilibrium sedimentation and n.m.r. sectroscopy. In contrast with a previous report [Smerdon & Isenberg (1976) Biochemistry 15, 4242--4247], it was found, by using equilibrium-sedimentation analysis, that protein HMG 1 binds to all three histone H1 subfractions CTL1, CTL2, and CTL3, arguing against there being a specific interaction between protein HMG 1 and only two of the subfractions, CTL1 and CTL2. Raising the ionic strength of the solutions prevents binding of protein HMG 1 to total histone H1 and the three subfractions, suggesting that the binding in vitro is simply a non-specific ionic interaction between acidic regions of the non-histone protein and the basic regions of the histone. Protein HMG 1 binds to histone H5 also, supporting this view. The above conclusions are supported by n.m.r. studies of protein HMG 1/histone H1 subfraction mixtures. When the two proteins were mixed, there was little perturbation of the n.m.r. spectra and there was no evidence for specific interaction of protein HMG 1 with any of the subfractions. It therefore remains an open question as to whether protein HMG 1 and histone H1 are complexed together in chromatin.


Changes associated with glycolytic-enzyme binding in the equatorial X-ray-diffraction pattern of glycerinated rabbit psoas muscle.

The binding of fructose biphosphate aldolase to the thin filaments of glycerinated rabbit psoas muscle produces a significant change in its low-angle X-ray-diffraction pattern. The intensity of the (11) reflection relative to that of the (10) reflection increases by 26 +/- 3% (mean +/- S.E.M.), which is consistent with the increase in the mass of the thin filaments produced by enzyme binding. A similar effect is found with a mixture of aldolase and glyceraldehyde 3-phosphate dehydrogenase. The significance of the change in intensity is considered with reference to the interpretation of the equatorial patterns obtained from muscles in different physiological states. The magnitude of the increase in the relative intensity of the (11) reflection is lower than that observed between relaxed and contracting muscle and does not bring into question the interpretation linking changes in these patterns to cross-bridge movement. However, the effect due to enzyme binding may be important when making detailed interpretations of these changes. It may also be related to an unusual pattern sometimes observed in cardiac muscle.


Purification and characterization of zinc-binding protein from the liver of the partially hepatectomized rat.

Zn-binding protein in liver of the partially hepatectomized rat was purified by column chromatography on Sephadex G-75 and DEAE-cellulose. Homogeneity was judged by polyacrylamide-disc-gel electrophoresis. The molecular weight determined by gel-permeation chromatography in 6 M-guanidine hydrochloride was 6700. This value is in good agreement with the molecular weight calculated from the amino acid composition, which was 6073. Zn-binding protein was composed of 61 amino acid residues, and the distinctive features include an extremely high content of cysteine, which accounted for one-third of the total amino acid residues, and an absolute absence of aromatic amino acids as well as of histidine, leucine and arginine. The amino acid composition was similar to that of the metallothioneins previously isolated from rat liver and mouse liver. These observations suggest that the Zn-binding protein can be classified as a type of metallothionein. Zn-binding protein contained 8.2g-atoms of zinc per mol and traces of copper, but no cadmium. The molar ratio of thiol groups to zinc was calculated to be 2.5:1. Possible roles of this Zn-binding protein in the transport and storage of zinc in the liver are discussed.


A new series of long-chain dicarboxylic acids with vicinal dimethyl branching found as major components of the lipids of Butyrivibrio spp.

1. Some members of the genus Butyrivibrio, including a general fatty acid auxotroph (strain S2), contain as a major part of their complex lipids a high-molecular-weight component that is probably formed by the union of two fatty acid chains [Hazlewood & Dawson (1979) J. Gen. Microbiol. 112, 15--27]. 2. Proton and 13C n.m.r. and i.r. and mass spectroscopy were used to examine a homologous series of these moieties and, in addition, the hydrocarbon derivative of one homologue and several synthetic compounds. 3. The results indicate that the high-molecular-weight components are a series of long-chain dicarboxylic acids containing vicinal dimethyl branching, located near the centre of the chain.


Hydrophobic proteins of lamellated osmiophilic bodies isolated from pig lung.

1. In addition to proteins that are insoluble in organic solvents, lamellated bodies isolated from pig lung and surfactant prepared from bronchopulmonary lavage fluid contain another group of proteins that are extracted together with lipid into the organic phase. 2. These hydrophobic proteins constitute about 40% of the total protein of lamellated bodies and about 13% of the total protein of surfactant isolated from lavage fluid, whereas less than 1% of the total protein of pig lung microsomal fraction and mitochondria is extracted by organic solvents. 3. The hydrophobic proteins of lamellated bodies were separated into four fractions and freed from phospholipid by chromatographic procedures. Their apparent molecular weights vary between 11 500 and 16 500, they contain 72--79% of hydrophobic residues and 16--22% of sulphur-containing amino-acids, and leucine is the major N-terminal amino acid in each case.


2,2'-Dithiodipyridine activates aldehyde dehydrogenase and protects the enzyme against inactivation by disulfiram.

Small concentrations of 2,2'-dithiodipyridine cause a rapid activation of sheep liver cytoplasmic aldehyde dehydrogenase in the presence of NAD+. Enzyme pre-modified by 2,2'-dithiodipyridine is largely protected against the potent inactivatory effect of disulfiram. 2,2'-Dithiobis-(5-nitropyridine) inactivates the enzyme. The implications of these results are discussed with reference to various possible classes of thiol group in aldehyde dehydrogenase.


Effects of fructose concentration on carbohydrate metabolism, heat production and substrate cycling in isolated rat hepatocytes.

1. Hepatocytes from starved rats were incubated with 5mm-glucose, labelled uniformly with (14)C and specifically with (3)H at positions 1, 2, 3 or 6, and with fructose at concentrations of 2.5, 7.5 or 25mm. 2. In the absence of other substrates only 1% of the radioactivity initially present in [U-(14)C]glucose appeared in the metabolic products, CO(2), lactate, pyruvate, amino acids and glycogen. 3. Fructose at 2.5mm caused a 30% increase in the glucose concentration and a 4-fold increase in the apparent oxidation of [U-(14)C]-glucose. 4. The formation of (3)H(2)O from [1-(3)H]-, [2-(3)H]-, [3-(3)H]- or [6-(3)H]-glucose was 2.4, 4.3, 2.15 or 1.6% respectively in the control incubations and 4.1, 10.4, 7.7 or 5.1% with 2.5mm-fructose. 5. Fructose at 7.5 and 25mm decreased the (3)H(2)O yields to less than the control values, but had no apparent effect on the amount of [U-(14)C]glucose metabolized. 6. In the incubations with 5mm-glucose and 25mm-fructose there were significant decreases in heat production, O(2) consumption and in the ratio of O(2) uptake to heat output. 7. Fructose at 2.5mm caused a 64% increase in heat output, but only a 43% increase in O(2) uptake. 8. The radioisotopic and calorimetric data demonstrate that physiological concentrations of fructose greatly increase metabolism in hepatocytes from starved rats. These data also indicate increased cycling at glucose/glucose 6-phosphate and at fructose 6-phosphate/fructose 1,6-bisphosphate in the presence of 2.5mm-fructose, although the rates of cycling were actually decreased relative to the amount of glucose catabolized. 9. At concentrations of 2.5, 7.5 and 25mm, fructose depressed hepatocyte ATP concentrations by 20, 65 and 80% respectively. Although fructose at 7.5 and 25mm increased glucose and lactate release, O(2) consumption, production of heat and formation of(3)H(2)O from [1-(3)H]-, [2-(3)H]-, [3-(3)H]- or [6-(3)H]-glucose were lowered to values equal to, or less than, controls. These effects probably reflect a severe derangement of hepatic metabolism due to excess phosphorylation of fructose when present at high concentrations.


Metabolic oxidation of the ethynyl group in 4-ethynylbiphenyl.

1. 4-Ethynylbiphenyl undergoes extensive metabolism in the rat and the rabbit, involving aromatic hydroxylation and oxidation of the ethynyl group. No metabolites containing the intact ethynyl group were detected. 2. In the rat unchanged 4-ethynylbiphenyl was concentrated initially in the adipose tissue. No other tissues accumulated significant amounts of radioactivity. 3. The major metabolites were the same in both the rat and the rabbit, namely 4'-hydroxybiphenyl-4-ylacetic acid (90-95% of dose) and biphenyl-4-ylacetic acid (2-10% of dose). 4. Excretion was slower in the rat than in the rabbit, probably because of greater biliary and faecal excretion in the rat. Biliary excretion and enterohepatic circulation of biphenyl-4-ylacetic acid and 4'-hydroxybiphenyl-4-ylacetic acid were demonstrated in the rat.


Hydrolysis of an exogenous 125I-labelled protein by rat yolk sacs. Evidence for intracellular degradation within lysosomes.

When added to the serum-free medium in which 17.5-day rat yolk sacs were incubated, formaldehyde-denatured 125I-labelled bovine serum albumin was rapidly degraded. More than 80% of the radiolabelled digestion products appearing in the incubation medium consisted of [125I]iodo-L-tyrosine; larger digestion products were found only in association with the yolk-sac tissue. In the early stages of an incubation, low-molecular-weight digestion products began to appear in the incubation medium only after they could be detected within the tissue, and progressive association of trichloroacetic acid-insoluble radioactivity with the tissue preceded both these events. None of the observed proteolysis could be attributed to proteinases released into the incubation medium. Tissue-associated acid-insoluble radioactivity showed a lysosomal distribution on sub-cellular fractionation, and cell-free homogenates of yolk sacs degraded albumin only at acid pH values. Progressively decreasing the rat of pinosome formation (either by progressively lowering the incubation temperature or by the use of increasing concentrations of the metabolic inhibitor rotenone) caused a corresponding decrease in the rate of degradation of albumin. These findings indicate that, in vitro, formaldehyde-denatured 125I-labelled bovine serum albumin is digested by rat yolk sacs exclusively intracellularly, within lysosomes.


Evidence for the rapid direct control both in vivo and in vitro of the efficiency of oxidative phosphorylation by 3,5,3'-tri-iodo-L-thyronine in rats.

1. Examination of the distribution of L-tri-iodothyronine among rat liver tissue fractions after its intravenous injection into thyroidectomized rats focused attention on mitochondria at very short times after administration. By 15 min this fraction contained 18.5% of the tissue pool; however, the content had decreased sharply by 60 min and even further over the next 3 h. By contrast, the content in all other fractions was constant or increased over 4 h. About 60% of tissue hormone was bound to soluble protein. 2. Mitochondria isolated from thyroidectomized rats showed P/O ratios that were about 50% of those found in normal controls, with both succinate and pyruvate plus malate as substrates. There was no evidence of uncoupling; the respiratory-control ratio was about 6. 3. Mitochondria isolated 15 min after injection of tri-iodothyronine into thyroidectomized rats showed P/O ratios and respiratory-control ratios that were indistinguishable from those obtained in mitochondria from euthyroid animals. The oxidation rate was, however, not restored. 4. Incubation of homogenates of livers taken from thyroidectomized animals injected with L-tri-iodothyronine before isolation of the mitochondria restored the P/O ratio to normal; by contrast, direct addition of hormone to isolated mitochondria had no effect. The role of extramitochondrial factors in rapid tri-iodothyronine action is discussed. 5. Possible mechanisms by which tri-iodothyronine might rapidly alter phosphorylation efficiency are considered: it is concluded that control of adenine nucleotide translocase is unlikely to be involved. 6. The amounts of adenine nucleotides in liver were measured both  after thyroidectomy and 15 min after intravenous tri-iodo-thyronine administration to thyroidectomized animals. The concentrations found are consistent with a decreased phosphorylation efficiency in thyroidectomized animals. Tri-iodothyronine injection resulted in very significant changes in the amounts of ATP, ADP and AMP, and in the [ATP]/[ADP] ratio, consonant with those expected from an increased efficiency of ADP phosphorylation. This suggests that the changes seen in isolated mitochondria may indeed reflect a rapid response of liver in vivo to tri-iodo-thyronine.


Synthesis of N-acetyl-L-aspartate by rat brain mitochondria and its involvement in mitochondrial/cytosolic carbon transport.

1. The synthesis and efflux of N-acetyl-l-aspartate from brain mitochondria of rats of different ages has been studied. 2. Brain mitochondrial State 3 (+ADP) respiration rate, using 10mm-glutamate and 2.5mm-malate as substrates, increases during the suckling period and reaches approx. 50% of the adult value at 17 days after birth [adult State 3 respiration rate=160+/-7ng-atoms of O/min per mg of mitochondrial protein(mean+/-s.d.; n=3)]. 3. The influence of 5mm-pyruvate or 10mm-dl-3-hydroxybutyrate on aspartate efflux from brain mitochondira from rats of different ages oxidizing glutamate and malate was studied. In all cases the aspartate efflux in State 3 was greater than in State 4, but, whereas the aspartate efflux in State 3 increased as the animals developed, that of State 4 showed only a small increase. However, the rate of aspartate efflux in the presence of pyruvate or 3-hydroxybutyrate as well as glutamate and malate was approx. 60-65% of that in the presence of glutamate and malate alone. 4. An inverse relationship between aspartate efflux and N-acetylaspartate efflux was observed with adult rat brain mitochondria oxidizing 10mm-glutamate and 2.5mm-malate in the presence of various pyruvate concentrations (0-5mm). 5. N-Acetylaspartate efflux by brain mitochondria of rats of different ages was studied in States 3 and 4, utilizing 5mm-pyruvate or 10mm-dl-3-hydroxybutyrate as acetyl-CoA sources. A similar pattern of increase during development was seen in State 3 for N-acetylaspartate efflux as for aspartate efflux (see point 3 above). Also only very small increases in N-acetylaspartate efflux occurred during development in State 4.6. Rat brain mitochondria in the presence of iso-osmotic N-acetylaspartate showed some swelling which was markedly increased in the presence of malate. 7. It is concluded that N-acetylaspartate may be synthesized and exported from both neonatal and adult rat brain mitochondria. It is proposed that the N-acetylaspartate is transported by the dicarboxylic acid translocase and may be an additional mechanism for mitochondrial/cytosolic carbon transport to that of citrate.


Acetate is the preferred substrate for long-chain fatty acid synthesis in isolated spinach chloroplasts.

1. Commercially available [2-14C]pyruvate and [2-14C]malonate were found to contain 3-6% (w/w) of [14C]acetate. 2. The contaminating [14C]acetate was efficiently utilized for fatty acid synthesis by isolated chloroplasts, whereas the parent materials were poorer substrates. 3. Maximum incorporation rates of the different substrates examined were (ng-atoms of C/h per mg of chlorophyll): [1-14C]acetate, 2676; [2-14C]pyruvate, 810; H14CO3-, 355; [2-14C]malonate, 19. 4. Products of CO2 fixation were probably not a significant carbon source for fatty acid synthesis in the presence of exogenous acetate.


Linoleate and alpha-linolenate synthesis by isolated spinach (Spinacia oleracea) chloroplasts.

Diacylgalactosylglycerol synthesis was a prerequisite for the incorporation of [1-14C]-acetate into linoleate and alpha-linolenate of isolated spinach (Spinacia oleracea) chloroplasts. Oleate at position 1 of diacylgalactosylglycerol was desaturated to linoleate and alpha-linolenate both in the light and in the dark. Some desaturation of palmitate was also observed after prolonged incubations.


The tricarboxylic acid cycle in Dictyostelium discoideum. Metabolite concentrations, oxygen uptake and 14c-labelled amino acid labelling patterns.

Some aspects of tricarboxylic acid-cycle activity during differentiation and aging in Dictyostelium discoideum were examined. The concentrations of glutamate, aspartate, alanine, citrate, 2-oxoglutarate, succinate, fumarate, malate, oxaloacetate, pyruvate and acetyl-CoA were determined at four stages over the course of differentiation. The rate of O2 utilization was also determined over differentiation. In addition, experiments are described in which the specific radioactivities of citrate, 2-oxoglutarate, succinate, fumarate and malate were determined during a 30 min labelling of cells from the preculmination stage of development with [14C]glutamate, [14C]aspartate or [14C]alanine. A similar experiment was also performed with cells from the aggregation stage of development using [14C]glutamate.


The tricarboxylic acid cycle in Dictyostelium discoideum. A model of the cycle at preculmination and aggregation.

A preliminary model of tricarboxylic acid-cycle activity in Dictyostelium discoideum is presented. Specific-radioactivity labelling patterns of intra- and extra-mitochondrial pools are simulated by this model and compared with the experimental data. The model arrived at by this method shows the following features. (1) The cycle flux rate is approx. 0.4 mM/min. (2) Both fumarate and malate are compartmentalized at approx. 1:5 between cycle pools and non-cycle pools. These may represent mitochondrial and cytoplasmic pools. Citrate is compartmentalized at 1:10. Succinate appears to exist in three compartments, two of which become labelled by [14C]glutamate and only one by [14C]aspartate (3) Two pools of aspartate with two associated pools of oxaloacetate are necessary for simulation. (4) Exchange between the cycle and non-cycle pools of both citrate and fumarate occurs at very low rates of about 0.003 mM/min, whereas exchange between the malate pools is about 0.004 mM/min. The exchange reaction glutamate in equilibrium 2-oxoglutarate runs at approx. 15 times the cycle flux. (5) A reaction catalysed by "malic" enzyme is included in the model, as this reaction is necessary for complete oxidation of amino acid substrates. (6) Calculation of the ATP yield from the model is consistent with earlier estimates of ATP turnover if the activity of adenylate kinase is considered.


A role for bicarbonate in the regulation of mammalian glutamine metabolism.

1. The concentration of HCO3- (independent of any change of pH) exerts different effects on glutamine metabolism in rat kidney-cortex tubules, hepatocytes and enterocytes.2. In kidney tubules HCO3- (10.5-50 MM) has no effect on glutaminase (EC 3.5.1.2), whereas glutamate dehydrogenase (EC 1.4.1.3) is inhibited as HCO3- concentration is increased. The result is that flux through the entire glutamate-to-glucose pathway is inhibited by increasing HCO3- concentrations. A large proportion (more than 30%) of the glutamine removed undergoes complete oxidation. 3. In hepatocytes, and to a smaller extent in enterocytes, HCO3- is an accelerator of glutaminase. Synthesis of glucose and urea from glutamine in hepatocytes increases as HCO3- concentration is increased. Calculations show that fumarate, formed via aspartate aminotransferase and arginino-succinate lyase, is the precursor of the glucose. There is no complete oxidation of the carbon skeleton of glutamine in hepatocytes. 4. Leucine at near-physiological concentrations (0.1-1 mM) is an accelerator of glutaminase in hepatocytes, but not in kidney tubules or in enterocytes. 5. The results are discussed in relation to regulation of acid/base balance in vivo.


Changes in the nuclear polyamine content of chick erythrocytes during embryonic development.

The polyamine content of the circulating erythrocyte population in the embryonic chick was studied during its development. Total cellular polyamine content fell dramatically between 5 and 7 days of development, paralleling the decrease in metabolic activity exhibited by these cells. Nuclei were isolated from the erythrocytes by a non-aqueous technique, which not only eliminated the polyamine loss that occurred with aqueous isolation, but also prevented redistribution of the polyamines from the cytoplasm. Nuclear spermidine and spermine contents decreased markedly between 5 and 6 days of development from 31 to 10 pmol/microgram of DNA and from 33 to 18 pmol/microgram of DNA respectively. Thereafter the spermine content remained constant, but the spermidine content continued to decline. Good correlations between spermidine and RNA contents were observed in both cells and nuclei, and similarly between spermine and RNA contents in cells, but no such correlation was observed between spermine and RNA in nuclei.


The degradation of cholic acid by Pseudomonas sp. N.C.I.B. 10590.

The microbial degradation of cholic acid by Pseudomonas sp. N.C.I.B. 10590 was studied, and two major products were isolated and identified as 7 alpha, 12 beta-dihydroxyandrosta-1,4-diene-3,17-dione and 7 alpha, 12 alpha-dihydroxy-3-oxopregna-1,4-diene-20-carboxylic acid. Four minor products were isolated and evidence is given for the following structures: 7 alpha, 12 alpha-dihydroxyandrosta-1,4-diene-3,17-dione, 12 beta-hydroxyandrosta-1,4,6-triene-3,17-dione, 7 alpha, 12 beta, 17 beta-trihydroxyandrosta-1,4-dien-3-one and 7 alpha, 12 alpha-dihydroxy-3-oxopregn-4-ene-20-carboxylic acid. The significance of the production of the steroid products is discussed, along with the possible enzymic mechanisms responsible for their production.


Identification of albumin as the plasma carrier for zinc absorption by perfused rat intestine.

The isolated vascularly perfused rat intestine exhibits an obligatory need for a protein carrier in order to absorb zinc. Therefore this system is ideal for use as a model to identify the plasma carrier during zinc absorption. Affinity chromatography on Blue Sepharose CL-6B was employed to separate the major serum zinc-binding proteins in the portal effluent of the perfused intestine. It  was found that 94% of newly absorbed 65Zn was transported in the portal serum-containing perfusate as an albumin-65Zn complex. The identity of albumin as the plasma carrier was confirmed by polyacrylamide-slab-gel electrophoresis. This evidence suggests that albumin is the plasma protein that is involved in removal of zinc from intestinal-mucosal cells and subsequent transport of the metal in portal blood to the liver.


Measurement of the protein-synthetic activity in vivo of various tissues in rats by using [3H]Puromycin.

The validity of a new technique was examined for estimating the protein-synthetic activity of various tissues in vivo. The basic assumption underlying the method is that the number of peptide chains growing on each active ribosome would increase as the protein-synthetic activity of each tissue increases. The principle of the procedure, which was devised originally by Wool & Kurihara [(1967) Proc. Natl. Acad. Sci. U.S.A. 58, 2401-2407] to determine in vitro the number of functional ribosomes in skeletal muscle, is as follows. Puromycin is known to bind easily to the C-terminal end of the growing peptide on ribosomes and thus stop further chain elongation. Hence, if the number of puromycin molecules attached to the nascent peptide is determined by using radioactive puromycin as a tracer, one can estimate the number of growing peptides, i.e. the activity of tissue protein synthesis. By using this technique, it is shown that both starvation and the feeding of a protein-free diet caused marked decreases in the relative rate of formation of peptidyl-puromycin, i.e. activity of protein synthesis in liver, skeletal muscle, heart, spleen, testis, lung, kidney and intestine.


Effect of hypophysectomy on liver nuclear ribonucleic acid synthesis in aging rats.

Changes in RNA synthesis in liver nuclei were observed at different ages and after hypophysectomy and hormone replacement in female Sprague-Dawley rats. As determined by the incorporation of [3H]UMP into an acid-insoluble product, RNA synthesis decreased by about 75% in intact rats from 6 months to 24 months of age. This decline with age was not observed in liver nuclei from 24-month-old rats that had been hypophysectomized at 12 months and maintained on a minimal hormone-replacement therapy. Thyroid hormones and somatotropin (growth hormone) had an additive effect on RNA synthesis in liver nuclei from these hypophysectomized rats. The same hormones had no significant effect on intact, age-matched rats. With advancing age, nuclei of intact rats had an increase in the pool of free RNA polymerase and an apparent decrease in the enzyme activity bound to nuclear chromatin. There was no change in total enzyme with age. In hypophysectomized, hormone-treated rats, free RNA polymerase activity decreased and chromatin-bound activity increased. There was no difference in total nuclear RNA polymerase activity between operated or intact rats. However, the ratio of the bound to the free activity was different. These results suggest that the ability of RNA polymerase to bind to chromatin may be involved in the age-related decrease in liver nuclear RNA synthesis of intact rats.


Secretion of haem by hepatic parenchymal cells.

Hepatic parenchymal cells in primary culture, and also the intact perfused liver, secrete newly synthesized haem into extracellular fluids. In cultures incubated with the haem precursor delta-amino[4-14C]laevulinate, labelled haem was formed at a linear rate for at least 8 h, and 10-20% of the total labelled haem was present in the culture medium. The appearance of labelled extracellular haem was proportional both to the concentration of labelled precursor offered to the cells and to the time of incubation. Similar results were obtained when [2-14C]glycine was added as haem precursor. Studies with the isolated perfused liver indicated that newly synthesized haem is secreted also by the intact liver. Approximately equal amounts of haem appeared in the bile and in perfusate. The findings are discussed in relation to the pathogenesis of symptoms in the hereditary hepatic porphyrias.


Prostaglandins and their precursors in tissues from rats fed on trans, trans-linoleate.

Feeding trans,trans-9,12-linoleate to rats as 50 and 100% of the dietary fat decreased the concentrations of n-6 fatty acids, i.e. 18:2, 20:3 and 20:4, in heart, kidney, lung, adipose tissue and platelets of rats. The concentrations of prostaglandin products prostaglandins E1, E2 and F2 alpha in serum were significantly decreased in rats receiving high concentrations of dietary trans, trans-linoleate.


Effect of calcium ions on ethanol oxidation and drug glucuronidation in isolated hepatocytes.

The effect of extracellular Ca2+ concentration on ethanol oxidation and drug metabolism was studied in isolated rat hepatocytes. Both ethanol oxidation and drug glucuronidation showed similar dependence upon Ca2+, which was a stimulation of activity as Ca2+ was increased to physiological concentration, and inhibition at higher concentration.


Fate of bradykinin-potentiating peptide 9a after intravenous injection.

The fat of less than Glu1-3H-labelled bradykinin-potentiating peptide 9a [BPP9a; less than Glu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro, an inhibitor of angiotensin-converting enzyme (peptidyl dipeptidase)] was studied in the rabbit. After intravenous injection, BPP9a was rapidly removed from blood and much of the associated radioactivity was excreted in urine. Approx. 8% of the radioactivity in urine collected 2h after drug administration occurred in the form of BPP9a itself, the remainder occurring in three lower homologues: less than Glu-Trp (60%), less Glu-Trp-Pro-Arg-Pro-Gln (20%) and less than Glu-Trp-Pro-Arg-Pro-Gln-Ile (12%). Hydrolysis was not accounted for by enzymes in blood or urine. Apparently hydrolysis occurred within the kidney, as less than Gl-Trp was obtained in 60% yield in urine of isolated rat kidney perfused with [less than Glu1-3H]BPP9a.


Effect of methylamalonic acid on gluconeogenesis in isolated rat and guinea-pig hepatocytes.

Gluconeogenesis from pyruvate, alanine, lactate and propionate was inhibited by methylmalonate in both rat and guinea-pig hepatocytes. The effect was dose-dependent. Gluconeogenesis from glycerol and xylitol was not affected.


Binding of Ricinus communis agglutinin to the mitochondrial inner membrane as an artifact during preparation.

Endosperm from Ricinus communis was homogenized in the presence of 3H-labelled Ricinus communis agglutinin, with or without addition of lactose. In preparations without the binding-specific sugar the subfraction containing the mitochondrial inner membrane contained sufficient labelled agglutinin to account for the agglutinin reported to be associated with this membrane.


[From ergot alkaloids to nicergoline/Chemical points of view (author's transl)].

The author reports the rationale of a line of research that has led to pharmacologically active ergoline derivatives from the more complex ergot alkaloids. Specific compounds showing oxytocic, alpha-adrenolytic and antiserotoninic activity were synthesized and the relationship between structure and activity discussed. Out of the compounds, 10-methoxy-1,6-dimethyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline) was chosen as the most interesting product for a detailed further development.


[Cerebral enzymatic activities related to energy transduction processes. A model for the evaluation of pharmacological changes in the brain of the adult rat].

A test model of studying the effects of chronic pharmacological treatment on cerebral metabolism related to energy transduction was developed. The most useful biochemical parameters were the cerebral enzymatic activities related to the glycolytic pathway (lactate dehydrogenase), the Krebs' cycle (citrate synthetase and malate dehydrogenase) and the electron transfer chain (total NADH-cytochrome c reductase and cytochrome oxidase). The model is based on the natural growth-dependent changes occurring in the rat during aging (from 10 to 60 weeks of life). As test drug, 10-methoxy-1,6-dimethyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion) was administered daily for three periods of 16 weeks each (10-26, or 28-44, or 44-60 weeks of life) by two different administration routes (oral and i.p.), and at two different dose levels: oral 1 or 4, i.p. 0.25 or 1 mg/kg. Biochemical data were obtained blindly after 4, 8, 12 and 16 weeks of treatment. The drug tested exerted different effects which were dependent on the various administration periods and the administration routes. No dose-effect relationship was established.


[Hemodynamic effects of nicergolin in man at rest and during exertion (author's transl)].

In 10 healthy subjects the injection of 10-methyl-1,6-diemthyl-ergoline-8 beta-methanol-(5-bromonicotinate (nicergoline, Sermion) (5 mg i.v.) caused a permanent decrease in blood pressure. This decrease in blood pressure is due to a rapidly occurring alpha-blocking effect and a secondary central alpha-stimulating effect which did not affect the postural regulating mechanisms of the arterial blood pressure in their function. The bradycardia and the elevation of the systolic volume and of the cardiac output are based on this central effect. After oral administration of 30 mg nicergoline the same modifications appeared but to a lesser extent. If nicergoline was administered in a dosage of 30 mg p.o. 1 h before bicyle exercise (100 W for 5 min), the cardiac output and blood flow in the lower extremities increased and the elevation of the systolic pressure was limited. However, the reactive tachycardia remained at the same level. The metabolic consequences of this administration resulted in an inhibited increase in the concentration of lactic acid in the blood, which is normally caused by this exertion. This favourable metabolic effect can be explained by 1. the initial bradycardia and the decrease in the total peripheral resistance whereby the oxygen consumption of the myocardium is reduced; 2. the peripheral vasodilation.


[Nicergoline and platelet aggregation. A review of experimental and clinical studies (author's transl)].

10-Methoxy-1,6-dimethyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion), an ergoline derivative in clinical use for syndromes related to cerebral and peripheral vascular insufficiency, displays a platelet antiaggregating effect which may be important for its therapeutic effect. This paper reviews the present experimental and clinical evidence relating to the platelet antiaggregating activity of nicergoline and its mechanisms of action is discussed in detail. Since the platelet antiaggregating effect of nicergoline is mainly related to its alpha-adrenolytic activity, the importance of catecholamines for the behaviour of both human platelets and endothelium as well as their interference with the prostaglandin/prostacycline system is also discussed.


[Effects of nicergoline on cerebral blood flow (author's transl)].

Cerebral blood flow (CBF) was measured before and after i.v. injection of the cerebral vasodilator 10-methoxy-1,6-dimethyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion) in 13 patients with cerebrovascular disease. CBF increased in seven. The possibility is discussed that the effect of the drug in the other patients may have been masked by a fall of CBF which occurs during sequential measurement of patients at rest. Nicergoline is an alpha-adrenergic receptor blocking agent which affects cerebral blood flow by reducing cerebrovascular resistance. The present study was undertaken to assess the acute effect of the drug on cerebral blood flow and blood pressure in patients with cerebrovascular diseases.


[Ultrastructural study on the effect of an inhibitor of platelet aggregation (author's transl)].

10-Methoxy-1,6-dimethyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion) is introduced into human platelet-rich plasma at different stages of collagen-, ADP- or epinephrine-induced aggregation. Ultrastructural fixation is processed while aggregation on the same plasma sample is recorded. If introduced before the aggregating agent, nicergoline completely neutralises its action and the platelets become spherical. The microtubule marginal bundle is disorganized and both open and dense canalicular systems are modified. If intoduced after the aggregating agent, nicergoline immediately stops the aggregation and disaggregation follows, with complete separation of the platelets. Morphology of microtubules and canalicular systems depend on the time before application of nicergoline. Nicergoline stops the induction of aggregation as well as ADP release. Disaggregation is an active process involving the microtubules.


[Physiopathology of the inner ear and therapy of presbyacusis (author's transl)].

Many experimental investigations have shown that labyrinthine fluids play a basic role in the physiology of the inner ear. Modifications of perilymphatic fluids have been demonstrated in perceptive deafness (otosclerosis, Ménière's disease, tympanolabyrinthosclerosis). Vasoactive agents fail to markedly affect the exchange between labyrinthine fluids, so that perceptive syndromes are generally regarded as unresponsive to treatment. Several studies have shown the effectiveness of 10-methoxy-1,6-diemthyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion) in various conditions of the inner ear. In this investigation, 30 patients with presbyacusis were treated with 30 mg/day p.o. for 30 days. Speech audiometric curves were improved in 4 out of 18 cases of physiological presbyacusis and in 6 out of 12 cases of accelerated presbyacusis. These results are very interesting, since few effective therapies are available to treat these conditions.


[Neurophysiological study of 36 patients with cerebral vascular processes treated with nicergoline (author's transl)].

The results obtained from a group of 36 patients with non-acute cerebral vascular disturbances of various forms were evaluated. The clinical course and additional examinations are reported: electroencephalography, visually evoked potentials and cerebral rheography for the assessment of different cerebral regions and an interhemispherical comparison. The determination of the data obtained by rheography can be applied to the whole patient group. All tests were carried out before treatment and after a period of treatment with 15-20 mg 10-methoxy-1,6-dimethyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion) administered daily p.o. On the basis of the therapeutic results conclusions can be drawn as to clinical improvement and EEG normalisation and visually evoked potentials. The changes in the amplitude and the changes in the systolic gradient recorded by the rheogramme as well as the tendency to correction of dyssymmetry can be described as the result of nicergoline on the cerebral vascular system, on the oxygen supply and on the metabolism of the brain tissue.


[Therapeutic efficacy of nicergoline in ophthalmology. Fluorescence retinographic study (author's transl)].

10-Methoxy-1,6-dimethyl-ergolin-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion) administered intravenously (8 mg) in a group of 10 healthy subjects caused a reduction of the arm-retina circulation time and a reduction of the ocular tone. The therapeutic effectiveness in 37 patients with various eye diseases was tested by administering nicergoline p.o. at a dose of 30 mg/day for 30 days and 20 mg/day as maintenance dose for 60-120 days. Positive results, in various percentages, were observed in cases of arterial obstructions, venous thrombosis, diabetic retinopathies, senile macular degenerations, papilla ischaemic oedema, central serous chorioretinopathies. Nicergoline caused no subjective or objective undesired effects.


Resorption of insoluble, heterologous, fluorescein-collagen sponges in sensitized and non-sensitized rats.

Sponges of insoluble bovine collagen were slowly resorbed over a 35-day period when implanted under the back skin of rats. The cellular picture was typical of a mild foreign-body reaction. The reaction to fluorescein-labelled collagen sponges was similar but there was evidence also of a weak immunological response. An acute inflammatory reaction with massive oedema was elicited when fluorescein-labelled collagen sponges were implanted in rats previously sensitized to either fluorescein-collagen or fluorescein-bovine serum albumin. The early invasion by PMN leucocytes subsided after 4 days and caused no observable breakdown of the sponge. The implanted material was rapidly encapsulated by fibrous tissue which was then resorbed along with the sponge between the 7th and 12th day. Macrophages were very active in the sponge at this time, sometimes forming giant cells. Fibroblasts were invading from the periphery with the development of the granulation tissue. The small residue which remained after this time was overrun by granulation tissue and was slowly resorbed up to the 35th day. Throughout the period of study there was only a weak local immunological response after the 28th day. The level of circulating antibodies against the fluorescein hapten was high, but the titre for the antibodies against bovine collagen remained low. The significance of these findings in the pathological destruction of connective tissue is discussed.


Premalignancy in vitro: progression of an initially benign epithelial cell line to malignancy.

An epithelial cell line, NMuLi, derived from normal mouse liver, produced a variety of tumours when inoculated s.c. into newborn mice. The inoculation of early-passage cells resulted in cystadenomas and proliferating cysts, while adenocarcinomas and carcinosarcomas were produced from cells that had been extensively subcultured. To determine the mechanism involved in the progression to malignancy, single-cell clones were isolated from early- and late-passage NMuLi cells. When inoculated into mice, early-passage clones were similar to the original population in that they initially formed benign cysts but produced increasingly aberrant tumours after extensive subculture. Inoculation of the late-passage clones resulted in only malignant tumours. These observations suggest that passage in culture induced an initially benign epithelial cell population to progress toward malignancy; thus early passage NMuLi cells represent a good in vitro model of the premalignant state.


Physiological alterations secondary to perfusion and revascularization of canine intestine.

Our studies suggest that the perfusion rates of canine intestinal segments during extracorporeal perfusion are directly related to the quality of preservation; namely, better preserved specimens exhibit higher perfusion rates and vice versa. Production of intraluminal fluid (ILF) during preservation and following revascularization is related to the quality of preservation. Poorly preserved intestine seems generally to produce more fluid than well preserved specimens. Conversely, continuous pulsatile perfusion techniques resulted in more ILF production than specimens perfused with low-flow gravity-dependent systems. Ex vivo intestinal perfusion resulted in organ oedema inversely proportional to the quality of preservation. Poor preservation is also associated with washout of potassium, lactic dehydrogenase, and tissue acidosis both during preservation and revascularization. Perfusion with a high concentration of potassium ions results in poor preservation presumably owing to vasoconstriction. Mucosal malperfusion as demonstrated by the Microfil technique is the circulatory abnormality most closely associated with inadequate preservation. Smooth-muscle function, as measured by electrical activity recordings, is well preserved by pulsatile flow methods, whereas gravity perfusion results in disorganized and spastic muscular activity. The best preservation appeared to be obtained by the use of pulsatile flow with cryoprecipitated plasma as the perfusate.


The migration of fibroblasts into an in vitro wound.

A method for the study in vitro of fibroblast migration in wounds is described. This consists of producing standard fibroblast monolayers (derived from subcultures of explants of flank skin from young male rats) in microtitre plates, wounding the monolayer in a linear fashion with razor blade, removing the monolayer from one side of the linear wound and retaining the other side for migration studies. Preliminary results using the method are described which illustrate its reproducibility and ease of performance.


The effect of indomethacin on the biochemical response to fracture stress in rats.

After acclimatization to metabolic cages, groups of male Wistar rats were pretreated for 48 h with parenteral indomethacin to observe its effect on the biochemical response to femoral fracture stress. This trauma induced an average 2.3-fold rise in fibrinogen levels in all animals, which was maximal 24 h after injury. Indomethacin pretreatment caused a dose-related decrease in the plasma fibrinogen response 24 h after fracture: 1, 2, 10 or 20 mg/kg/day of drug reduced the response by approximately 3, 10, 13 and 23%. Fibrinogen levels in rats treated with high doses (10 and 20 mg/kg) of drug began to rise before fracture owing to intestinal ulceration. Plasma 11-hydroxycorticosteroids and urine total catecholamine levels did not respond to either indomethacin treatment or to fracture but levels of both compounds rose gradually during the experiment. Indomethacin is an inhibitor of prostaglandin synthesis and prostglandins may therefore be involved in the metabolic response to stress.


Dissociation of erythema and basophil accumulation in Jones-Mote type hypersensitivity in the guinea-pig.

Relationship between delayed-in-onset erythema and infiltrating leucocytes in the skin reaction site of Jones-Mote type hypersensitivity was observed in guinea-pigs. Guinea-pigs were immunized with formalin-fixed (F-SRBC) or native form (N-SRBC) of sheep red blood cells in incomplete Freund's adjuvant (IFA). Elicitation was carried out with intradermal injection of F-SRBC or N-SRBC in saline 1 or 3 weeks after immunization. One week after immunization with F-SRBC or N-SRBC in IFA, erythema accompanied by basophil infiltration was detected, but antibody production was negative. Erythema reached a peak at 24 h, but basophil infiltration reached a peak at 48 h or later. In the skin reaction site elicited with N-SRBC, high levels of basophil infiltration were detected persistently even after erythema had disappeared. Three weeks after immunization, low titres of PCA (passive cutaneous anaphylaxis) were detected in guinea-pigs immunized with F-SRBC in IFA. At that time erythematous skin reaction was detected, but the level of basophil infiltration was lower than that of 1 week after immunization. In guinea-pigs immunized with N-SRBC in IFA, skin reaction of the Arthus type accompanied by haemorrhage was observed at the site elicited with N-SRBC.


Effects of intravenous injection of two different strains of Corynebacterium parvum in the mouse.

A strain of C. parvum, CN6134, known to have antitumour activity, caused thrombosis in the sites where the organism is phagocytosed. It bound to macrophages in vitro and activated the alternate pathway of complement. A strain of C. parvum, CN5888, which fails to show antitumour activity, did not show thrombosis. It did not bind to macrophages or activate guinea-pig complement. It did, however, cause marrow infarction which seems to result from a diminished clearance of the organism from the circulation.


Histochemical studies on peroxisomes in regenerating proximal tubules of the kidney.

Peroxisomes of the regenerating proximal tubules of the rat kidney were investigated after necrosis induced by mercuric chloride. Slices both for light and electron microscopic examinations were incubated in 3,3'-diaminobenzidine (DAB) medium. Peroxisomes were absent in the necrotic epithelium. They appeared on the fourth day of regeneration and later their number increased reaching the normal distribution in the fourth week. They seem to be needed for the functional differentiation of the proximal tubule cells during regeneration.


The histopathology and ultrastructure of pleural mesotheliomas produced in the rat by injections of crocidolite asbestos.

Primary tumours of the pleural cavity were produced in rats by the intrapleural injection of crocidolite asbestos. Their histological structure as seen with both light and electron microscopy was very variable and tumours frequently contained elements of both connective-tissue and epithelial type. In some instances the connective-tissue elements predominated from the start and the earliest tumour nodules consisted mainly of pleomorphic connective-tissue cells with only a few layers of cells more nearly epithelial in type on the surface. This pattern was largely retained when tumour nodules increased in size and coalesced, but in the deeper layers of advanced tumours the pleomorphic connective-tissue pattern was often replaced by a more uniform spindle-cell form. Other tumours were more predominantly epithelial in type, showing either a papillary pattern with rounded epithelial cells growing in solid columns, or a vesicular form in which large tissue spaces, often intracellular, were lined by very thin layers of extended cell cytoplasm. Whereas early tumours showed only one histological pattern, the more advanced stages often exhibited areas of all 3, so that there seemed to be some degree of histological mutability. The spindle-cell areas of advanced tumours frequently showed evidence of direct invasion of the surrounding tissue but this was never seen with the epithelial forms of rat mesothelioma.


A study of the effect of povidone-iodine on polymorphonuclear leucocyte chemotaxis.

Studies were made of the effect of povidone-iodine on polymorphonuclear leucocyte chemotaxis. Polymorphonuclear leucocytes were incubated with various concentrations of povidone-iodine and allowed to migrate across a membrane towards a chemotactic agent. Chemotactic movement was found to decrease as the concentration of povidone-iodine rose, 75 micrograms/ml completely inhibiting all movement. A concentration of 10 micrograms/ml of povidone-iodine was found actively to repel the white cells. In vivo studies in mice showed a reduction in polymorphonuclear leucocytes at wound surfaces in the presence of povidone-iodine dry powder spray.


[Sulfate transport across the limiting double membrane or envelope, of spinach chloroplasts].

Evidence is presented for low rates of carrier-mediated uptake of sulphate, thiosulphate and sulphite into the stroma of the C3 plant Spinacia oleracea. Uptake of sulphate in the dark was followed using two techniques (1) uptake of sulphate [35S] as determined by silicon oil centrifugal filtration and (2) uptake as indicated by inhibition of CO2-dependent O2 evolution rates after addition of sulphate. Sulphate, thiosulphate and sulphite were transported across the envelope leading to an accumulation in the chloroplasts. Sulphate for each molecule of sulphate entering the chloroplast, one molecule of phosphate leaves the stroma, and vice-versa. The uptake of sulphate by isolated intact chloroplasts exchanging for internal free phosphate induced a lower rate of photophosphorylation, which in turn inhibited CO2-dependent O2 evolution. The presence, on the inner membrane of the chloroplasts envelope, of a specific sulphate carrier, distinct from the phosphate translocator, is discussed.