Extraparticulate chain interaction between different electron transport particles.

Interaction or cross-linking between the respiratory chains of the electron transport particles of bacterial origin occurs with a mixture of active and inactive particles. Interaction between bacterial particles and liver sub-mitochondrial particles also occurs. Irradiation of the bacterial particles at 360 nanometers resulted in the destruction of quinone and consequent loss of ability of reduced nicotinamide adenine, dinucleotides to reduce cytochromes b, c(1), c, and a plus a(3). A mixture of both irradiated and untreated particles in the presence of the reduced dinucleotide resulted in the reduction of cytochromes c and a plus a(3), in an amount equivalent to the total concentration of these cytochromes in both types of particles. In contrast, the amount of cytochrome b reduced was equivalent to half the particle concentration or to that observed with the active particles alone. The rate of reduction of cytochromes c and a plus a(3) with the mixture of particles was similar to that with the active particles alone. The interaction or cross-linking between the particulate respiratory chains of bacteria or of bacterial and mammalian systems occurs after cytochrome b and before or at cytochrome c.

Effect of parathyroid hormone on rat heart cells.

Myocardiopathy is common in uremia, but its cause in unknown. Excessive entry of calcium in heart cells by catecholamines has been shown to cause necrosis of myocardium. The high blood levels of parathyroid hormone (PTH) in uremia may also enhance entry of calcium into heart cells and exert deleterious effects on the heart. We examined the effect of PTH on rat heart cells grown in culture. Both amino-terminal (1-34) PTH and intact (1-84) PTH, but not the carboxy-terminal (53-84) PTH produced immediate and sustained significant rise in beats per minute and the cells died earlier than control. The effect was reversed if PTH was removed from medium, and was abolished by inactivation of the hormone. There was a dose-response relationship between both moieties of PTH and the rise in heart beats, but the effect of 1-84 PTH was significantly greater than that of 1-34 moiety. PTH stimulated cyclic AMP production within 1 min, and cyclic AMP remained significantly elevated thereafter. The effect of PTH required calcium, was mimicked by calcium ionophore, was prevented by verapamil and was not abolished by alpha- or beta-adrenergic blockers. PTH action was additive to phenylephrine and synergistic with isoproterenol. Sera from uremic parathyroidectomized rats did not effect heart beats, but sera from uremic rats with intact parathyroid glands or from uremic-parathyroidectomized rats treated with PTH had effects similar to PTH. Data indicate that (a) heart cell is a target organ for PTH and may have receptors for the hormone; (b) PTH increases beating rate of heart cells and causes early death of cells; (c) PTH effect appears to be due to calcium entry into heart cells; (d) the locus of action through which PTH induces calcium entry is different from that for catecholamines; and (e) uremic serum has no effect unless it contains PTH. Data suggest that myocardial damage may occur in uremia due to prolonged exposure to very high blood levels of PTH, and assign new dimensions to PTH toxicity in uremia.

Effect of parathyroid hormone on osmotic fragility of human erythrocytes.

The survival of erythrocytes (RBC) is shortened in uremia, and it has been shown that calcium influx into RBC evoked crenation and increased their rigidity. The high blood levels of parathyroid hormone (PTH) may augment entry of calcium into RBC and hence affect their integrity. We examined the effect of PTH on osmotic fragility of human RBC and investigated the mechanisms through which PTH interacts with RBC. Both the amino-terminal (1-34) PTH and the intact (1-84) PTH, but not the carboxy-terminal (53-84) PTH, produced significant increases in osmotic fragility. This effect was abolished by prior inactivation of the hormone. There was a dose-response relationship between both moieties of PTH and the increase in osmotic fragility. This action of PTH required calcium, was mimicked by calcium ionophore, and was partially blocked by verapamil. PTH caused significant influx of (45)Ca into RBC, which was not associated with potassium leak. The hormone did not affect water content of RBC. Scanning electron microscopy revealed that the incubation of RBC with PTH was associated with the appearance of membrane filamentous extensions, which anchor RBC together. Inhibition of glycolytic activity of RBC with NaF or inhibition of Na-K-activated ATPase with ouabain did not abolish the effect of PTH on osmotic fragility. PTH did not stimulate RBC Na-K-activated ATPase or Mg-dependent ATPase but caused marked and significant stimulation of Ca-activated ATPase. The basal activity of the RBC adenylate cyclase was low and PTH produced only a modest stimulation of this enzyme. Both cyclic AMP and dibutyryl cyclic AMP had no effect on osmotic fragility. THE DATA INDICATE THAT: (a) the RBC is a target organ for PTH, (b) the hormone increases osmotic fragility of RBC, and (c) this effect of PTH is due to enhanced calcium entry into RBC. We suggest that the increased calcium influx may affect the spectrin-actin of the cytoskeletal network of the RBC and may alter the stability and integrity of the cell membrane. This action of PTH on the RBC could be, at least in part, responsible for the shortened survival of RBC in uremia, and assign a new role for PTH in the pathogenesis of the anemia of uremia.

Effect of parathyroid hormone on erythropoiesis.

Inhibitors of erythropoiesis have been found in the blood of uremic patients but their nature has not been identified. These patients have excess blood levels of parathyroid hormone (PTH) and it is possible that PTH inhibits erythropoiesis. The present study was undertaken to examine the effect of intact PTH molecules and some of its fragments on human peripheral blood and mouse bone marrow burst-forming units-erythroid (BFU-E), on mouse bone marrow erythroid colony-forming unit (CFU-E), and granulocyte macrophage progenitors (CFU-GM), and evaluate the interaction between PTH and erythropoietin (Ep) on human BFU-E. Intact PTH (1-84 bPTH) in concentrations (7.5-30 U/ml;) comparable to those found in blood of uremic patients produced marked and significant (P less than 0.01) inhibition of BFU-E and mouse marrow GFU-GM, but not of mouse marrow CFU-E. Inactivation of 1-84 bPTH abolished its action on erythropoiesis. Increasing the concentration of Ep in the media from 0.67 to 1.9 U/ml overcame the inhibitory effect of 1-84 bPTH on BFU-E. The N-terminal fragment of PTH (1-34 bPTH) and 53-84 hPTH had no effect on BFU-E. The results demonstrate that (a) either the intact PTH molecule or a C-terminal fragment(s) bigger than 53-84 moiety exerts the inhibitory effect on erythropoiesis, and (b) adequate amounts of Ep can overcome this action of PTH. The data provide one possible pathway for the participation of excess PTH in the genesis of the anemia of uremia.

Biochemical changes associated with the osmotic fragility of young and mature erythrocytes caused by parathyroid hormone in relation to the uremic syndrome.

The effect of parathyroid hormone at concentrations found in uremic patients on erythrocytes (RBC) from newborn and adult rabbits was studied in relation to the fragility pattern in hypotonic salt solutions and the activities of Ca- and Mg-dependent ATPases. Median osmotic fragility of RBC from newborn rabbits was significantly lower than in mature rabbits. Parathyroid hormone (PTH) stimulated to a greater extent the mean osmotic fragility in RBC from newborn rabbits, than in those from adults. Similarly, the hormone stimulated to a much greater extent the Ca-ATPase but not the Mg-ATPase in RBC from the newborn rabbits, in comparison to those from adult rabbits. PTH, which is greatly elevated in the blood of patients with chronic renal failure, may be one cause of the anemia seen in these patients, and its effect, which is mediated by Ca-ATPase activity, is stronger on young RBC. There were significant morphological changes in the young RBC caused by PTH, as seen with scanning electron microscopy.

Effect of parathyroid hormone and uremic sera on the autoagglutination and sedimentation of human red blood cells.

Parathyroid hormone (PTH) caused a dramatic acceleration of erythrocyte sedimentation rate (ESR). This effect was calcium dependent and was partially reversed by verapamil. It was not mimicked by 5 mumol/l calcium ionophore A-23187. Following the removal of PTH from the cell suspension the ESR returned to normal. PTH also caused haemagglutination, the reaction was Ca2+ dependent, pH dependent and was partially reversed by verapamil. High levels of Ca2+ ionophore A-23187 mimicked this phenomenon. Magnesium ions even at concentrations of 5 mmol/l did not replace Ca2+, while Ca2+ at concentrations of 3 mmol/l and above caused haemagglutination. The glycolytic inhibitor NaF at levels of 1 mmol/l did not inhibit haemagglutination. The polyamines pertusin and spermidin, prostaglandins PGE2 and PGF, and the calcium hormone calcitonin, did not reproduce the PTH effect. Dialysate from serum of patients with chronic renal failure and hyperparathyroidism caused haemagglutination, while dialysate from patients with chronic renal failure following parathyroidectomy and normal individuals did not cause this phenomenon. It seems that abnormal erythrocyte behaviour seen in patients with chronic renal failure is caused by PTH which leads to modified Ca2+ metabolism in these cells.

Biochemical changes in liver, kidney and blood associated with common bile duct ligation.

Common bile duct ligation (CBDL) in rats was used to induce liver disease and secondary kidney damage. The biochemical changes in the liver, kidney and plasma were studied at 3, 6, 10 and 21 days post CBDL. The observed alterations climaxed at the 6th day following ligation. Renal, activities of aldolase (ALD), lactic dehydrogenase (LDH), isocitric dehydrogenase (ICDH), sorbitol dehydrogenase (SDH), and alkaline phosphatase (ALP), were lowered in CBDL rats. Further, microsomal Na,K-ATPase and Mg-ATPase and mitochondrial oxidative-phosphorylation were inhibited. In the liver from CBDL rats the activities of aspartate aminotransferase (AST), Mg-ATPase and ALP were elevated, while SDH, ALD, malic dehydrogenase (MDH), LDH, malic enzyme (ME) and Na,K-ATPase were lowered. Plasma enzymes, AST, ALP, MDH, LDH, ALD, acid phosphatase (ACP) and ICDH and the metabolites bile acids, bilirubin, creatinine and urea were elevated. Addition of bile acids or bilirubin at concentrations comparable to those found in the plasma of CBDL rats, to the reaction mixture of the various enzymes strongly inhibited most, particularly mitochondrial oxidative phosphorylation. High concentrations of these substances in the blood may explain the development of renal failure during liver disease and its reversibility when liver function returns to normal.

Effect of parathyroid hormone and uremia on erythrocyte deformability.

Parathyroid hormone (PTH) caused a significant decrease in human erythrocyte filtration rate (EFR). This effect was Ca2+-dependent and was partially reversed by the Ca2+ blocker verapamil. It was mimicked by the Ca2+ ionophore A-23187. Mg2+ even at high concentrations could not substitute for Ca2+. There was a dose response between the filtration rate and Ca2+ or PTH concentrations. Serum ultrafiltrate of patients with chronic renal failure and secondary hyperparathyroidism caused significant inhibition similar to that seen with PTH extract. Ultrafiltrate from patients with chronic renal failure following parathyroidectomy and from healthy individuals did not cause this phenomenon. Erythroyctes deformability in uremic patients appears to be caused by high PTH levels. Our present findings agree with ones about the toxic effects of PTH on various biological systems.

Changes in renal enzyme activities following the administration of gentamicin to unilateral nephrectomized rats.

The effects of unilateral nephrectomy and the impact of gentamicin administration on renal tissue enzyme activities in adult Wistar rats were investigated. Gentamicin 200 mg/kg body wt. or an equivalent volume of saline to control rats was administered subcutaneously on three consecutive days, followed by unilateral nephrectomy. Rats were killed on day 3, 7 or 14 following nephrectomy. Alkaline phosphatase, predominantly a proximal tubular brush border enzyme, rose in both the experimental and control groups, however, significantly less in the gentamicin treated rats. Aspartate aminotransferase activity, an enzyme participating in renal glucogenesis, increased transiently in the control but remained unchanged in the experimental group. No difference in glucose-6-phosphate dehydrogenase activity between the two groups was observed, probably reflecting the localization of this enzyme to distal tubular segments, a site unaffected by gentamicin. Significant and similar increases in Mg2+ and Na+ K+ ATPase were observed on day 14 in both groups. The administration of the drug resulted in a marked reduction in oxygen consumption, with a higher oxidation to phosphorylation ratio (P/O). Serum creatinine concentration was significantly higher on days 3 and 7 in the experimental group reverting to control values on the 14th day. Urea concentration increased significantly on days 3 and 7, decreasing on the 14th day to values slightly, but significantly, higher than those of the controls.

Selective neurotoxicity induced by the ionophore lasalocid in rat dissociated cerebral cultures, involvement of the NMDA receptor/channel.

An in vitro model of dissociated cerebral cultures, prepared from prenatal 15-16-days rat fetuses, was used to further characterize the neurotoxic effects caused by the antibiotic ionophore lasalocid-X-537A. The damage caused by lasalocid (1-2 microM, 2-4 hr) included swelling of perikarya, followed by cytolysis of most neurons present in the cultures. The neuronal damage was dose-dependent, noticeable at concentrations above 0.5 microM, and was more pronounced in established cultures (14 days in vitro-DIV) than in younger ones (7 DIV). Unlike neurons, no damage was observed in glia and other non-neuronal cells present in the cultures by exposure to 2 microM lasalocid. Moreover, the drug was not toxic for cultures of rat astrocytes and C6 glioma cells. Another calcium ionophore A-23187 (calcimycin, 1 microM), destroyed both neuronal and non-neuronal cells within 1 hr. Ca2+ influx was increased by 140% in cultures exposed to lasalocid (1.5 microM). The lasalocid neurotoxic effects were neither inhibited by 10 microM nimodipine (a calcium channel antagonist) nor by 10 microM 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX)(a non-N-methyl-D-aspartic acid (NMDA) receptor antagonist), but were exclusively blocked by 10 microM MK-801 (a non-competitive NMDA receptor/channel antagonist). The neurotoxicity induced by lasalocid was further confirmed by measurements of lactate dehydrogenase (LDH) released into the media. Lasalocid (1.5 microM) induced the release of both LDH and arachidonic acid (AA) (by 8 and 4 fold of control values, respectively), and this was blocked by MK-801 but not by CNQX. These results are in according with the observations that activation of calcium influx through the NMDA receptor leads to activation of phospholipase A2 (PLA2) and release of AA. In contrast, MK-801 did not block the release of either LDH or AA mediated by the calcium ionophore A-23187 (1 microM) in these cultures. [3H]-MK-801 binding to washed rat cortical membranes, a measure of direct interaction with the NMDA receptor/channel complex, was not affected by lasalocid either alone or in the presence of glutamate and glycine. [3H]-D-aspartate release, a measure of excitatory amino acid (EAA) secretion mediated by NMDA receptor activation, was increased by lasalocid and could be blocked by MK-801. These observations suggest that lasalocid induces selective neurotoxicity, which involves the NMDA receptor/channel complex, possibly indirectly, resulted in elevated intracellular Ca2+ levels and the subsequent glutamate or aspartate release.

Gamma-glutamyltransferase as a marker for the pasteurization of raw milk.

The degree and rate of inactivation of gamma-glutamyltransferase in raw cow's milk by heating at 50, 60, 70, and 80 degrees C for 1, 2, 3, 5, 10, 15, 20, 25, and 30 min were measured to evaluate the suitability of this enzyme as a marker for the pasteurization of milk. The enzymes alkaline phosphatase and lactate dehydrogenase were also measured under similar conditions for comparison. The patterns of heat inactivation of gamma-glutamyltransferase and alkaline phosphatase were similar, with only a minimal inactivation of the enzymes at 50 degrees C. The rate of inactivation increased as a result of increasing temperatures and time. A complete inactivation of both enzymes was seen at 70 degrees C after 10 min and at 80 degrees C after 1 min. Lactate dehydrogenase showed a higher heat resistance with almost complete inactivation at 70 degrees C for 30 min, and compete inactivation at 80 degrees C for 3 min. No activities of these enzymes were found in commercially pasteurized or heat-treated milk. The levels of gamma-glutamyltransferase in raw milk were between 8 and 10% higher than those of alkaline phosphatase and lactate dehydrogenase, making it more sensitive and accurate as a testing marker. It seems that gamma-glutamyltransferase may serve as a good pasteurization marker. Furthermore, the simplicity of testing and the availability of commercial kits for testing by both wet and dry chemistry make it an attractive choice, especially because dry chemistry procedures overcome the difficulties originating from the turbidity of milk, which interferes with spectrophotometric procedures.

Buffalo-milk enzyme levels, their sensitivity to heat inactivation, and their possible use as markers for pasteurization.

The activities and rates of inactivation of four enzymes in raw buffalo milk were measured in relation to the process of heating to determine the value of these enzymes as markers for the evaluation of milk pasteurization. The activities of the enzymes alkaline phosphatase (ALP), lactic dehydrogenase (LDH), gamma-glutamyltransferase (GGT), and aspartate aminotransferase (AST) were measured before and after heating at 50, 60, 70, and 80 degrees C for 1, 3, 5, 10, 20, and 30 min. The enzyme GGT showed the highest activity (712 +/- 601 IU/liter), followed by LDH (386 +/- 183 IU/liter), ALP (295 +/- 164 IU/liter), and AST (18 +/- 4 IU/liter). Heating the milk at 50 degrees C for 1 to 30 min resulted in no effect on the activity of any of the enzymes. At 60 degrees C, ALP showed the highest sensitivity to heat inactivation, whereas all other enzymes showed resistance. At 70 degrees C, ALP activity was abolished completely after 1 min, whereas GGT and LDH lost most activity after 10 min, and AST still maintained 50% activity even after 30 min. At 80 degrees C, the activities of LDH and GGT were lost, whereas AST still retained some of its activity. The results suggest that in addition to ALP, LDH and GGT, but not AST, are potential markers for heat denaturation in buffalo milk, with GGT having the advantage that its concentration is the highest.

Evaluation of buffalo colostrum quality by estimation of enzyme activity levels.

A study to evaluate the value and potential use of colostral enzymes as markers for the evaluation of buffalo colostrum quality was conducted. The enzymes gamma-glutamyltransferase (GGT), lactic dehydrogenase (LDH), and alkaline phosphatase (ALP) in buffalo's colostrum were measured spectrophotometrically, and their activities were correlated with the gamma-globulin content. Gamma-globulin concentration was determined following the electrophoretic separation of the colostral proteins and quantified with a densitometer. Colostrum was obtained from 15 dams, soon after calving. Means, standard deviations, correlation coefficients, and degree of significance were calculated using the general linear model procedure of the Statistical Analysis Systems program. The activity of GGT in the colostrum was the highest, followed by LDH and ALP. A significant correlation (r = 0.86; P < 0.001) was seen between GGT and gamma-globulin concentration in the colostrum, supporting the suggestion of using this enzyme as a marker for the evaluation of colostrum quality.

Serum protein alterations in canine ehrlichiosis.

Serum protein electrophoresis was performed in 42 dogs with naturally occurring Ehrlichia canis infection and in 15 clinically healthy dogs (control dogs). The infected dogs were found to have a significant hypoalbuminaemia, hyperglobulinaemia and hypergammaglobulinaemia compared to the control dogs (P < 0.001). A polyclonal gammopathy was found in all but one of the infected dogs which presented a monoclonal gammopathy. alpha-1 globulin was lower while alpha-2 and beta-2 globulin concentrations were significantly higher in the infected dogs (P < 0.0001, P < 0.05 and P < 0.005, respectively). The infected dogs were divided into two subgroups according to haematological parameters, defined as pancytopenic (n = 13) and non-pancytopenic (n = 29). When compared, the pancytopenic group revealed significantly lower concentrations of total protein, total globulin and gammaglobulin (P < 0.01, P < 0.05 and P < 0.005 respectively). The lower concentrations of the gammaglobulins coupled with the pancytopenia suggest that the immune state of the pancytopenic E. canis infected dogs is more compromised, and therefore secondary infections should be expected more frequently in these dogs.

Characterization of the subclinical phase of canine ehrlichiosis in experimentally infected beagle dogs.

Beagle dogs were examined during the subclinical phase of canine ehrlichiosis under controlled conditions. Emphasis was placed on gathering data before artificial inoculation with Ehrlichia canis, and comparing these data with those of the subclinical phase of the disease. In this study all dogs were clinically healthy throughout the 6 month examination period. All subclinically infected dogs had IFA antibody titers to E. canis at a dilution varying from 1:2560 to 1:20480. The most prominent haematological finding was mild thrombocytopenia with a concomitant increase in platelet size, seen in eight of the nine dogs examined. Leukocyte counts were statistically significantly reduced in 78% of the dogs, compared with their preinfection values, with 71% of dogs having significantly reduced absolute neutrophil counts. None of the dogs were either leukopenic nor neutropenic. Six of the nine dogs had increased serum gamma-globulin concentrations. No dogs were overtly anemic, although declines in packed cell volume, haemoglobin concentration and total erythrocyte count were detected in an inconsistent manner among the dogs. It was concluded that, the most reliable parameters for judging possible subclinical ehrlichial infection in beagle dogs was mild thrombocytopenia, together with a persistently high antibody titer to E. canis. Hypergammaglobulinemia would increase the suspicion further. Based on the results presented, routine testing of dogs in E. canis endemic areas is recommended in order to identify and treat dogs in the subclinical phase of the disease.

Biochemical changes associated with the fatty liver syndrome in cows.

There were significant changes in enzyme activities and concentrations of metabolites in the blood and liver of cows with fatty livers when compared to normal cows. Blood and liver samples were taken from cows at the abattoir immediately after slaughter. The liver was checked for pathological signs and the samples were divided according to the degree of fatty changes. Three groups were studied: controls showing no gross pathological signs, mild fatty infiltration and severe infiltration. In cows with fatty liver, there were significant increases in the serum activities of isocitric dehydrogenase (ICDH), glucose-6-phosphate dehydrogenase (G6PDH), glutamic dehydrogenase (GLDH), lactic dehydrogenase (LDH), malic dehydrogenase (MDH), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and acid phosphatase (ACP). In the fatty liver, the activities of the enzymes, ICDH, G6PDH, LDH, MDH, ALP and malic enzyme (ME) were significantly higher, while sorbitol dehydrogenase (SDH) was significantly lower. While serum total lipid decreased, the opposite was seen in the liver with higher lipid content, mainly due to triglycerides and cholesterol esters. The significant increases in the NADPH generating enzymes ME, ICDH, G6PDH and MDH, which are required for fatty acid synthesis, suggest that the lipids accumulated in the liver are not only of extrahepatic origin, mobilized into the liver, but also arise from increased lipid synthesis in the liver which is induced during the laying down of fat in the liver. Measurement of the serum NADPH generating enzymes may serve as a useful biochemical test specific for fatty liver in cows.

Biochemical changes in fowl serum during infection with strains of Newcastle disease virus of differing virulence. Changes in serum proteins, uric acid, lipids and electrolytes.

Chickens aged five to six weeks were inoculated with three strains of Newcastle disease virus of differing pathogenicity. The serum level of the metabolites: total protein, albumin, globulin, uric acid, total lipids, cholesterol and electrolytes: calcium, magnesium, sodium and potassium were determined. The changes in serum levels of metabolites were as follows: velogenic infection was accompanied by decrease in total protein and albumin, reduction of lipids and cholesterol and increase in uric acid. No significant changes were found in the values of these metabolites in the serum of chickens infected with mesogenic strain. Lentogenic strain caused elevation of uric acid and cholesterol. All three strains caused decrease of the level of potassium in serum.

Lactate dehydrogenase isoenzyme distribution and patterns in chicken organs.

Unlike most mammals, chicken lactate dehydrogenase isoenzymes cannot be separated using the 'Titan-Gel' electrophoresis. However, using isoelectric focusing at a pH range of 3.0 to 9.0, a good and clear separation of all five isoenzymes was achieved. Generally, three characteristic groups were seen: (a) those having a cathodic domination (breast muscle and serum) with mainly lactate dehydrogenase-5 (b) those having an anodic domination (heart, muscle, liver, pancreas, kidney, erythrocytes) of mainly lactate dehydrogenase - 1 and 2 and (c) those with a more uniform distribution (spleen, lung, and brain). The total lactate dehydrogenase activity was the highest in the breast muscle, followed by the heart muscle, liver and serum with the lowest activities in the lung and pancreas.

Pharmacokinetic changes of several antibiotics in chickens during induced fatty liver.

The concentrations of five antibiotics (erythromycin, lincomycin, penicillin G, streptomycin and oxytetracycline) were determined in chicken serum before and after induced fatty liver. The pharmacokinetic variables were calculated according to the obtained data. The crossover trial design involved 10 chickens for each antibiotic. The fatty liver was produced by oestradiol-dipropionate injections and monitored by serum malic enzyme activity determinations. Protein binding of the respective antibiotics was determined in vitro in the serum obtained from normal and oestrogen-treated birds. Induction of fatty liver caused several changes in the determined variables. The measured peak concentrations were higher for lincomycin and erythromycin and lower for penicillin and oxytetracycline while streptomycin remained unchanged. The peak concentration of streptomycin appeared earlier and the peak of oxytetracycline later than in the normal chickens. The elimination half-lives were shorter for erythromycin, lincomycin and streptomycin and increased for penicillin and oxytetracycline. The area under the concentration curve (AUC) decreased for erythromycin, penicillin and streptomycin, increased for oxytetracycline and remained unchanged for lincomycin. The body clearance (ClB/f) and the apparent specific volume of distribution (Vd(area'/f) were considerably changed in association with fatty liver induction. Since the fraction of the drug absorbed (f) is not known, it can only be speculated that changes in distribution rather than reduced liver function altered the kinetics. The protein binding was decreased for all the antibiotics, but this did not seem to be the reason for changes in kinetics, except perhaps in the case of penicillin.

Distribution of lactate dehydrogenase isoenzymes in normal and inflamed bovine udders and milk.

The patterns of distribution of lactate dehydrogenase (LDH) isoenzymes were determined in normal and inflamed bovine udder tissues, in normal and mastitic milk-leucocytes and serum. LDH1 was the most common isoenzyme found in all the types of tissues examined, normal tissues contained the lowest proportions of LDH5 whereas the inflamed tissues and leucocytes from mastitic milk showed a higher proportion of LDH4 and LDH5. It seems that the origin of the elevated LDH in mastitic milk is the leucocytes and the parenchyma cells of the udder. The significance of the shift in the molecular forms of LDH isoenzymes is discussed in relation to possible alternations in energy metabolism in the inflamed bovine mammary gland.