Serum calcium and magnesium in leprosy.

Serum calcium and magnesium were studied in 70 leprosy patients and 25 normal healthy individuals. An attempt has been made in this study to find out if there is any correlation between the clinical and pathological status of the disease and serum calcium and magnesium levels in the blood. Serum calcium was found to be significantly decreased in lepromatous leprosy (Ca=8.42+/-0.7 mg%, P less than 0.001) and dimorphous leprosy (Ca=8.68+/-0.94 mg%, P less than 0.05) while it was normal in tuberculoid leprosy (Ca=9.14+/-2.12, P less than 0.05). The decrease in serum magnesium level was highly significant in all clinical types of leprosy (Mg=1.08+/-0.29, P less than 0.001).


Enzyme histochemistry of the sheep uterus during the oestrous cycle.

Enzyme histochemical techniques were applied to frozen sheep uteri from different stages of the oestrous cycle. The localization and activities of succinate, lactate, glucose-6-phosphate, and isocitrate (NADP+) dehydrogenases and acid and alkaline phosphatases were studied in the luminal and glandular epithelia, caruncle and myometrium. Enzyme activity in the sections was scored on a scale of 0--5. In general the enzyme activity in the uterine caruncles and epithelia was higher than in the myometrium. The myometrium did not show any alkaline phosphatase activity and isocitrate dehydrogenase (NADP+) activity was negligible. The low activities of acid phosphatase and lactate dehydrogenase and the moderate levels of glucose-6-phosphate and succinate dehydrogenases in the myometrium were constant. The caruncular tissue showed high levels of phosphatases and glucose-6-phosphate dehydrogenase, moderate levels of lactate and succinate dehydrogenases, and low levels of isocitrate dehydrogenase (NADP+) throughout the oestrous cycle. Much lower phosphatase and isocitrate dehydrogenase (NADP+) levels were found in the epithelium of deep glands compared with superficial glands. The high activity of acid and alkaline phosphatases in the luminal epithelium and the superficial glands was constant from mid-cycle to ovulation, but a significant decrease was observed immediately after ovulation. The level of dehydrogenases in epithelia was generally high and did not change during the oestrous cycle.


Ethanol extraction of basic proteins from ejaculated human spermatozoa.

A method for the extraction of basic proteins from human ejaculated spermatozoa has been developed. It relies on the previously unreported observation that such basic protein is soluble in a solution containing 60% (v/v) ethanol. This unconventional method yields a high percentage of arginine-rich basic protein which is then able to be characterized on the basis of its amino acid composition. This method also allows comparisons to be made between single ejaculates by the banding pattern each displays when subjected to polyacrylamide gel electrophoresis.


Haemoglobin oxidation in whole blood samples from sheep in relation to glutathione peroxidase activity.

A high correlation was observed between H2O2-induced oxidation of haemoglobin and glutathione peroxidase activity in whole blood samples from sheep. A role of this enzyme in the prevention of oxidative damage to the erythrocyte and its contents has been previously demonstrated. The possibility of using haemoglobin oxidation in whole blood as an alternative assessment of glutathione peroxidase activity and hence of selenium status is proposed.


Optimal conditions for lactoperoxidase catalyzed radioiodination of external proteins on mouse erythrocytes.

Optimal conditions were established for specific labelling of the surface proteins of mouse erythrocytes using lactoperoxidase-catalyzed radioiodination. The levels of H2O2 and I-, and cell concentrations required for restriction of haemoglobin labelling to less than 5% of the total 125I-protein, were different for radioiodination employing direct H2O2 addition or generation of H2O2 with glucose oxidase plus glucose. Preparation of mouse erythrocyte ghosts by hypotonic lysis caused loss of some minor labelled proteins present on intact cells and shifts to lower molecular weights of others. It is therefore important to solubilize labelled cells directly in electrophoresis buffer to avoid artifactual degradation of labelled proteins. The extent of labelling internal cell proteins was measured by a procedure suitable for the comparison of a large number of samples: solubilized radioiodinated erythrocytes were electrophoresed on 14% acrylamide gels and the radioactivity determined in the haemoglobin band which migrates separately from other proteins. The major labelled protein on the mouse erythrocytes had an apparent molecular weight of 92,000, and may be analogous to Band 3 of the human erythrocyte.


IgG1 hypergammaglobulinaemia in chronic parasitic infections in mice: magnitude of the response in mice infected with various parasites.

Mice chronically infected with 3 metazoan and 1 protozoan parasite contain in their circulation levels of IgG1 which are increased over the levels in uninfected mice by at least 10x. In the case of infection with the larval cestode, Mesocestoides corti, the serum IgG1 concentration can reach greater than 50 mg/ml and, with a half-life of less than 2 days, the number of cells engaged in IgG1 production is approximately 2 x 10(8). The IgG1 hypergammaglobulinaemia is not seen in infected hypothymic nude mice. Biosynthetic labelling studies with organ and tissue cultures established that in two of the chronic infections the organs principally involved in IgG1 synthesis were those pathologically involved or those "in line" for antigen capture: i.e. liver and spleen in the case of M. corti which is located in the liver and the peritoneal cavity, and various intestinal lymph nodes in the case of the gut-dwelling nematode, Nematospiroides dubius. This apparently exaggerated response to chronic parasitic infection is of interest simply because of the potential magnitude of the effect and the fact that it involves an Ig isotype with very poorly defined biological function.


Possible pathways of contamination of meat and bone meal with Salmonella.

Samples taken at various points along the processing line in 2 rendering plants showed that post-processing contamination occurs almost immediately after termination of the heat process in the percolator and surge bin. Further contamination occurs at each processing stage. In plant A 12.5% of the samples collected from material leaving the surge bin, 36% of the samples of product after it has passed the milling stage and 61% of samples collected from the stored product were found to be contaminated with salmonellas. In plant B the results were 15%, 40% and 69% respectively. Heavy contamination of product left overnight in percolators and surge bins, which are not cleaned routinely, was considered an important source of early post-processing contamination. The ecology was found to be similar at both plants despite the fact that in one plant the uncooked and cooked areas were separated completely, whilst this was not the case in the other plant. None of the air samples collected yielded salmonellas, while nearly all the insect samples collected in the rendering plants yielded salmonellas.


Brucella suis infection in pregnant cattle.

Six pregnant, Bos taurus cows with stages of gestation ranging from 11 to 33 weeks were each inoculated into the right conjunctival sac with 0.2 ml of a smooth culture of Brucella suis type I containing 27 x 10(6) viable organisms. The 6 cows produced 7 calves of which one single calf and one twin calf were stillborn, the cause of which was not determined. Br. suis was not isolated from any of the cows or calves using either special media or guinea pig inoculation. No abnormality was found in any of the cows or calves at autopsy. Microscopic examination of placentas and tissues from stillborn calves revealed no abnormality. Serologically, 2 weeks after inoculation all 6 cows had positive reactions to the Rose Bengal Test (RBT) and serum agglutination (SAT) titres of 25 iu to 116 iu. However, these reactions disappeared within 11 weeks. Only 2 cows had a complement fixation (CFT) titre which lasted a maximum of 5 weeks and reached a titre of 4/4. Following the anamnestic use of Br. abortus strain 45/20 vaccine on 3 of the cows, positive RBT reactions, SAT titres of 33 iu, 29 iu and 58 iu and CFT titres of 4/16, 1/8 and 3/8 respectively were recorded 6 weeks after vaccination.


Bruising in cattle fasted prior to transport for slaughter.

Three trials, each with 100 bullocks previously grazing improved pasture, compared bruising in cattle fasted for 0, 24 and 48 hours prior to transport for slaughter. Overall the weight of bruised trim from unfasted bullocks (0.48 kg) was significantly less (P less than 0.01) than from bullocks fasted for 24 h (0.99 kg) or 48 h (1.03 kg). length of fast overall had no significant effect. This pattern was not consistent within trials.


The effectiveness of levamisole hydrochloride in the treatment of adult Dirofilaria immitis.

Dogs infected with D. immitis were treated with oral levamisole hydrochloride at various dose rates over different durations. Clinical toxicity, haematological and biochemical parameters, body weight and food intake were recorded and filarial mortality estimated at autopsy. All filariae were dead in dogs treated at 10 mg/kg body weight twice daily (12 hourly) for 14 days, with minimal clinical toxicity being seen during the treatment period.


Y chromosome morphology of cattle.

Metaphase chromosomes from cultured lymphocytes were prepared from 246 bulls including Bos indicus, Bos taurus. Bos (Bibos) banteng, Sanga and interspecific and intra-specific breed crosses. Morphology and karyotype position of the Y chromosome for each bull were noted. Karyotype position of the Y chromosome varied between bulls from 25th to 29th pair and the Y chromosomes of Bos indicus and breeds derived from Bos indicus bulls were acrocentric while those of Bos taurus, Sanga and breeds derived from these bulls were metacentric/submetacentric. Two forms of Y chromosome were noted in the Droughtmaster breed. C-banding patterns of the acrocentric Y chromosome were characteristic and enabled easy identification.


The epidemiology of ringworm in racehorses caused by Trichophyton equinum var autotrophicum.

The epidemiology of dermatomycocis due to Trichophyton equinum var autotrophicum was studied in a number of thoroughbred stables in south-east Queensland. The significant factors in the epidemiology were defined. The infection was readily transmitted, particularly by infected saddle-girths, on which the fungus could survive for 12 months. Mild abrasion from the saddle during work favoured the development of lesions and prolonged the recovery period. A pronounced age and seasonal incidence of the disease was demonstrated as young horses under the age of 3 years were most susceptible, and the majority of cases occurred in periods of high humidity.


Some hatchery factors involved in early chick mortality.

Observations made in a commerical broiler hatchery revealed that chicks hatched over a period of 48 hours. Chick mortality to 10 days of age was 3.2% for those hatched at the commencement of the hatch, 1.2% for those hatched at peak of hatch and 52.9% for those hatched at the end of hatching. Chicks hatched early were more prone to dehydration while late hatching chicks had a higher incidence of leg weakness. Chicks held for 48 hours in hatcher machines lost 12.5% to 21.7% of their hatching weight and 79.4% of the hatching weight of the yolk sac. Normal 10-day mortality from this hatchery in winter months was observed to be 2.4% but was reduced to 1.2% when staggered setting times of donor flocks was employed by removing chicks from the machines 3 hours after 100% hatch, but was increased to 5.6% by holding chicks in the hatchery in chick boxes for 24 hours at 70 degrees C.


The incidence of thiabendazole resistance in field populations of Haemonchus contortus on the northern tablelands of New South Wales.

A survey was undertaken to assess the incidence of thiabendazole resistance in field populations of H. contortus in sheep flocks on 40 farms distributed throughout the sheep grazing area of the northern tablelands of New South Wales. Treatment with thiabendazole at 66 mg/kg reduced H. contortus ova output by 96-100% in 18 flocks while in the remaining 22 there was a range of responses down to levels where the anthelmintic was without apparent effect. Factors which may contribute to the high incidence of anthelmintic resistance are discussed and the possibility of widespread resistance occurring in other sheep raising areas is considered.


Pneumonia associated with Bordetella bronchiseptica in captive koalas.

Bordetella bronchiseptica was isolated from 12 cases of purulent bronchopneumonia from a captive koala colony near Brisbane, Queensland. An initial epizootic in March 1967 causing 13% mortality was followed by annual outbreaks causing 2% mortality usually in newly weaned and aged koalas in late winter to early spring. High population density and a low plane of nutrition in winter were thought to predispose to the occurrence of the disease.


Eperythrozoon ovis -- a cause of anaemia, reduced production and decreased exercise tolerance in sheep.

Sheep infected with E. ovis have lowered Hb and PCV values, reduced weight gains, reduced wool production and decreased exercise tolerance when compared with uninfected sheep.


Efficacy of albendazole against Fasciola hepatica in sheep.

In series of 4 trials dose rates of 3.8, 4.75 and 7.5 mg/kg of albendazole were given to sheep either as a single or repeat dose such as may be given under field conditions. Efficacy against 6-to 7-week-old Fasciola hepatica ranged from 16.0% to 43.6%. Efficacy against mature 12- to 17-week-old fluke ranged from 74.4% to 94.5%.


Melioidosis in wallabies.

This paper records 2 fatal cases of melioidosis in wallabies from Australia maintained in the Botanical Gardens, Penang, for about 3 months. The nature of lesions described is different from that described by various workers.


Medroxyprogesterone acetate and reproductive processes in male dogs.

The treatment of normal male dogs with a depot preparation of medroxyprogesterone acetate (4 mg/kg) for 7 weeks reduced peripheral testosterone levels by 58%. No effects on testicular size and consistency, semen quality or libido were found.


[On the determination and metabolism of diethylallylacetamide (author's transl)].

A report on the paper, thin-layer and gas chromatographical determination of diethyl-allyl-acetamide and its principal metabolites in urine specimens. Chemical and physical examinations show that diethyl-allyl-acetamide decomposes in the organism to 2,2-diethyl-4-hydroxymethyl-lambda-butyrolactone.


Urinary metabolites of clomethiazole. Detection and structural analysis by gas chromatography-mass spectrometry.

As the result of a renewed extensive investigation of clomethiazole (Distraneurin) metabolism five previously unknown metabolites could be isolated from human urine. Their structures were elucidated by mass spectrometry. Whereas previous investigations on the metabolism of clomethiazole had demonstrated changes only of the ethyl group, we now found metabolites attached to the methyl group, too. The newly isolated compounds 5-(1-hydroxy-2-chloroethyl)-4-methylthiazole (9) and 5-(2-hydroxyethyl)-4-thiazole carboxylic acid lactone (5) were found to be more abundant in human urine than 4-methyl-5-thiazole acetic acid previously considered as the main metabolite.


A novel class of potent topical antiinflammatory agents: 17-benzoylated, 7 alpha-halogeno substituted corticosteroids.

As part of continued efforts in the synthesis of structurally novel corticosteroids, a number of 17 alpha-benzoylated, 7 alpha-halogeno substituted prednisolones were tested for topical antiinflammatory activity. 7 alpha-Chloro, 7 alpha-bromo, and 7 alpha-iodo corticosteroids were synthesized by hydrogen halide addition to 1,4,6-triene-3-ones. The 7 alpha-fluoro substituted steroid was obtained by reaction of the appropriate 7 beta-hydroxy compound with N,N-diethyl(2-chloro-1,1,2-trifluoroethyl)amine. Antiinflammatory potencies were obtained using a croton oil-induced inflammation in the ears of mice. In this assay the greatest effect of a 7 alpha-halogen was observed in the 16 alpha-methylprednisolone series, where 7 alpha-fluoro and 7 alpha-bromo substitution yielded corticosteroids with topical potencies significantly higher than those of the corresponding 17,21-dipropionate analogs. Surprisingly, little potency enhancement due to 7 alpha-halogenation was discerned in the 16 beta-methylprednisolone series.


Gas chromatographic determination of verapamil in plasma and urine.

A gas-liquid chromatographic method for the determination of verapamil in body fluids has been developed. Verapamil and internal standard are extracted from alkalized fluid with heptane and then back extracted into 1 N HCl. After re-extraction into heptane, verapamil and internal standard are analysed by gas-liquid chromatography using a nitrogen-specific detector (N-FID). The method is specific for verapamil. Concentrations can be measured down to 4 ng ml-1 plasma. The coefficients of variation in plasma samples were 5--8% (within-day; range of concentration 5--30 ng ml-1). Recovery in this range was 100 +/- 6.2% during several weeks.


Synthesis and anti-inflammatory properties of budesonide, a new non-halogenated glucocorticoid with high local activity.

As part of a study of the local anti-inflammatory activity of corticosteroid 16 alpha, 17 alpha-acetals it was found that on acetalization of 16 alpha-hydroxyprednisolone with n-butyraldehyde the two possible epimers were formed in the ratio of 1: 1. The reaction product was resolved by column chromatography on Sephadex LH-20. The isolated epimers were studied by NMR and mass spectrometry. The epimeric mixture of this new non-halogenated corticoid, 16 alpha, 17 alpha-(22R,S)-propylmethylenedioxypregna-1,4-diene-11 beta,21-diol-3,20-dione (budesonide), was shown to have a local antiinflammatory potency comparable to that of fluocinolone acetonide in cotton pellet tests in rats. In contrast, its systemic glucocorticoid activity was found to be 4--7 times lower than that of fluocinolone acetonide, however.


A high pressure liquid chromatographic method for the determination of tetracyclines in blood and organs of experimental animals.

A high-pressure liquid chromatographic method has been developed for the quantitative analysis of tetracyclines from biological material. Tetracyclines can be extracted from serum or organ homogenates by means of acetonitrile containing buffer. The extract is chromatographed on a reversed-phase column and the tetracyclines are monitored spectrophotometerically at 357 nm. Tetracyclines, pyrrolidinomethyl-tetracycline, 4-epitetracycline, and doxycycline are thus separated within 8 min. The lowest concentration for the intact molecule detectable are 0.2 micrograms/ml serum or blood and 0.4 micrograms/g organ. In a comparative study the new extraction procedure and the chromatographic analysis yielded the same results as a microbiological method.


Distribution of pyrrolidinomethyl-tetracycline (rolitetracycline) and tetracycline in blood and various organs of mice measured by high pressure liquid chromatography.

By means of a newly developed high-pressure liquid chromatographic method the organ distribution of tetracycline (TC) and pyrrolidinomethyl-tetracycline (PMT) has been studied. When mice were treated with 50 mg/kg i.v. TC or PMT these antibiotics could be detected in all organs investigated (liver, kidney, heart, lung, muscle, spleen). Especially high concentrations were found in liver and kidneys, where TC and PMT could be detected up to 6 h. In animals treated with PMT part of the PMT applied decomposed slowly yielding TC, which was found together with PMT in blood and all organs.


[On the influence of clanobutine on the formation of restraint ulcer in rats (author's transl)].

Prophylactic effect of 4-[(4-chlorobenzoyl)(4-methoxyphenyl)-amino]-butyric acid (clanobutine, Bykahepar) on restraint stress ulcer formation was studied in male albino rats. Number and size of ulcers were counted, pH value of gastric juice and plasma levels of corticoids, glucagon and blood sugar were measured. Rats treated with clanobutine had only the one-third of ulcers compared with the untreated animals. This effect was found to be dose-dependent. Clanobutine did not change plasma levels of corticoids, glucagon or blood sugar. Rise in pH value of gastric juice during stress exposition was found to be lower in clanobutine treated animals than in untreated controls. Blood flow in gastrointestinal tract was seen to be much better under the influence of clanobutine. The mechanism of action of clanobutine is discussed.


Metabolism and kinetics of renal elimination of N-picolyl-3,5-dimethylbenzamides.

The metabolism and kinetics of renal elimination of N-(2-picolyl)-, N-(3-picolyl)- and N-(4-picolyl)-3,5-dimethylbenzamides were studied. These products have a depressive activity on the central nervous system, as well as an anti-inflammatory and spasmolytic activity. The elimination of the products N-(3-picolyl)- and N-(4-picolyl)-3,5-dimethylbenzamides includes a process of metabolic oxidization of the atom of pyridinic nitrogen with formation of the corresponding N-oxides as main products of its bio-transformation. The process of N-oxidization was not observed with the product N-(2-picolyl)-3,5-dimethyl-benzamide. The results showed that the renal elimination rate of the products studied is greater as the pyridinic nitrogen moves away from the amide group. In the biotransformation process of these products, there is an opposite effect, i.e., the formation of corresponding N-oxides is lower as the pyridinic nitrogen approaches the amide group.


Normalizing effects of S-methylmethionine (vitamin U) on lipoprotein disorder in rats.

S-Methylmethionine (vitamin U) dose-dependently lowered the increased plasma LDL and normalized the decreased HDL in dietary-induced hyperlipidemic rats.


[Effect of behavior changing drugs, imipramine and tranylcypromine, on the composition of the blood and activity of the alkaline leukocytophosphatase of emotive and non-emotive adult male Wistar rats].

The corpuscular composition of the blood and the alcalic leucocytophosphatase (ALP) activity was measured for a total of 40 emotive and non-emotive adult male Wistar rats in order to test the influence of a single dose of imipramine (IMI) and tranylcypromine (TRAN) as well as of IMI and TRAN given in a period of 8 days. Special attention was paid to the reaction type of the animals. Changes in the red and white components of the blood were found due to drug application and to reaction type. The relationship between reaction type and drug application we assumed to exist was confirmed. The ALP activity also changed depending on the type of drug dosage. The assumption that ALP activity is higher for emotive rats than for non-emotive ones was confirmed as well.


Pharmacokinetics of a fixed combination with propranolol, hydrochlorothiazide and triamterene.

The blood levels of propranolol, hydrochlorothiazide and triamterene, three components of a new antihypertensive combination, Dociteren, were studied in patients with mild hypertension after single dosage. The blood levels of propranolol in the combination did not differ from the blood levels obtained after a single dose of propranolol given alone.


[Pharmacokinetic studies on broad spectrum antibiotic combinations of penicillin and oxacillin in normal volunteers (author's transl)].

After i.v. application of broad spectrum penicillins (ampicillin, mezlocillin, carbenicillin) alone or combined with oxacillin (concomitantly) in different dosages to normal volunteers, the course of serum levels and recovery from urine were assessed and pharmacokinetic data were calculated for a 1-compartment model, in some cases also for a 2-compartment model. The following penicillins were tested alone and combined with oxacillin; ampicillin 3 g, oxacillin 2 g and their combination; ampicillin 2 g, oxacillin 1 g and their combination; mezlocillin 3.75 g, oxacillin 1.25 g and their combination; mezlocillin 2 g, oxacillin 1 g and their combination; carbenicillin 7.5 g, oxacillin 2.5 g and their combination. Apart from one combination (oxacillin 2 g + ampicillin 3 g), all penicillin combinations exceeded the serum levels achieved after individual application at any time. In combinations, the elimination constants decreased, total clearance was lower, and half-lives increased. The distribution volumes varied. With one exception, oxacillin 2.5 g + carbenicillin 7.5 g, all the combinations yielded a higher recovery from urine than did individual administration. When administering i.v. two penicillins simultaneously, a process similar to that following administration of probenecid in association with caronamide must need be considered. It may be assumed that the prolongation and elevation of serum levels after combined administration likewise lead to higher tissue concentrations, which effect might be of clinical consequence.


[Quantitative EEG examinations on the vigilance stabilizing effect of nicergoline / Results of a double blind study with gerontopsychiatric patients (author's transl)].

A randomized, double blind, cross-over study was carried out on long-term gerontopsychiatric patients. The aim of the study was to compare the EEG effects of a four weeks' treatment with 10-methoxy-1,6-dimethyl-ergoline-8 beta-methanol-(5-bromonicotinate) (nicergoline, Sermion) with those of dihydroergotoxin-mesylate (DHETM) and a four weeks' placebo period. The results of the power spectrum analysis with a consecutive factor analysis of the spectral data show that nicergoline and DHETM have a vigilance stabilizing effect which is characterized by a decrease in the relative power of slow delta-theta frequencies and an increase in the alpha power. As opposed to DHETM, for which this effect could not be proved, nicergoline produces a significant increase in power of the fast alpha frequencies as well as of the beta 1, beta 2, and beta 3 band.


Studies on the antibacterial activity of two new acylureidopenicillins, mezlocillin and azlocillin.

The antimicrobial activity of 6-[(R)-2[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenylacetamido]-penicillanic acid sodium salt (mezlocillin, Baypen) and 6-[(R)-2-(2-oxo-imidazolidine-1-carboxamido)-2-phenylacetamido-a1-penicillanic acid sodium salt (azlocillin, Securopen) was measured against 545 clinical isolates, including gram-negative rods, gram-positive cocci and Bacteroides. Mezlocillin was more effective than azlocillin against the majority of the strains studied, but azlocillin was more effective against Pseudomonas strains. The minimal bactericidal concentration was equal to the minimal inhibitory concentration for the strains tested, but it was twice or four-fold as high for Staphylococcus.


Synergistic action between sisomicin and mezlocillin against gram-negative bacteria and Staphylococcus aureus.

The combined effect of sisomicin and 6-[(R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido-a1-penicillanic acid sodium salt (mezlocillin, Baypen) was studied against 50 bacterial strains, including Pseudomonas aeruginosa, Proteus spp. Klebsiella-Enterobacter, E. coli and Staphylococcus aureus. No antagonism or indifference was detected with the strains studied. Both antibiotics were synergistic against 62% of the strains, and partially synergistic against 38%. Out of the bacteria studied, Staphylococcus aureus was the most susceptible to the combined action of sisomicin and mezlocillin.


[High pressure liquid chromatographic analysis of azlocillin and its penicilloate in urine].

A high-performance liquid chromatographic method has been developed for the determination of 6-[(R)-2-(2-oxo-imidazolidine-1-carboxamido)-2-phenyl-acetamido]-penicillanic acid sodium salt (azlocillin, Securopen) and its major metabolite the azlocillin-penicilloic acid in human urine. The separation of the parent compound and its metabolite from the interfering material of the urine was performed by gradient elution technic using reversal-phase material as stationary phase. Urine was diluted with phosphate buffer, filtered through a micropore membrane and the filtrate was injected directly onto the chromatographic column.


[Pharmacokinetics of Azlocillin in premature and newborn infants (author's transl)].

In 13 newborns and 12 prematures the pharmacokinetic parameters of 6-[(R)-2-(2-oxo-imidazolidine-1-carboxamido)-2-phenyl-acetamido]-penicillanic acid sodium salt (azlocillin, Securopen), were investigated after a single i.v. load of 50 mg/kg body weight. From the concentration-time curve an open two-compartment model could be postulated. The values of the microconstants indicate that there is a rapid diffusion from the peripheral compartment into the central one. The elimination half-life calculated from the beta-slope is 2.6-2.5 h. differences between newborns and prematures are lacking. To reach an average steady-state concentration C-infinity between 50 and 80 microgram/ml plasma, 100-200 mg azlocillin/kg body weight during 24h must be given. The accumulation rates for a dosage interval of 6 or 8 h are 0.6 and 0.45.


[On the excretion of azlocillin with bronchial secretion in children (author's transl)].

This pharmacokinetic study involved school children who were treated with 6-[(R)-2-(2-oxo-imidazolidine-1-carboxamido)-2-phenyl-acetamido]-penicillanic acid sodium walt (azlocillin, Securopen) at doses of 50 mg/kg bodyweight or 75 mg/kg body weight i.v. Kinetics of the first order were found for the concentrations measured at the same time, being proportionate to the administered dose, while there was no dose dependence for the relative distribution volume. The elimination half-life was about 55 min. In 25 children of different age who had to undergo bronchoscopy for diagnositic and therapeutic reasons, the azlocillin concentrations in the bronchial secretions were measured 1 h and 4 h after the administration of 75 mg/kg quantity of bronchial secretion was obtained, azlocillin concentrations of 30 - 240 microgram/g were found. Measurements of azlocillin concentration in the bronchial secretion and in the serum, which were made at the same time, did not show any correlation.


Pharmacokinetics of azlocillin in children with cystic fibrosis.

6-E1(R)-2-(oxo-imidazolidine-1-carboxamido)-2-phenyl-acetamido]-penicillanic acid sodium salt (azlocillin, Securopen) was given in doses of 100 and 200 mg/kg body weight to children with cystic fibrosis. After intravenous bolus infections, the serum half-life was 0.82 +/- 0.12 h after the lower dose and 0.98 +/- 0.18 h after the higher dose. This was consequent to a dose limited elimination kinetics due to limitation in both renal and non-renal processes of elimination. Upon doubling of the dose from 100 to 200 mg/kg, the total body clearance dropped from 13.93 to 5.10 1/h. Evaluation of data presented in other publications indicate that a dose limited elimination kinetics is the normal situation for azlocillin. Besides, the serum concentrations of patients with cystic fibrosis were considerably lower than seen in the healthy state. The reason is faster elimination by the renal route in cystic fibrosis.


Comparison between the pharmacokinetic properties of mezlocillin when given intravenously as a bolus injection or as an infusion.

6-[(R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido]-penicillanic acid sodium salt (mezlocillin, Baypen) in doses of 5.0 g was given i.v. as an infusion or as a bolus injection to 5 healthy volunteers. 5 min after the completion of the infusion, the mean serum concentration was 430 +/- 81 microgram/ml compared to 424 +/- 91 microgram/ml after the bolus injection. The half-lives were 1.31 +/- 0.24 h and 1.72 +/- 0.21 h, respectively. These results indicate that differences in serum concentrations between various investigators probably can be attributed to differences in body size and do not seem to depend on the administration route.


Elimination kinetics of mezlocillin in normal and impaired renal function including the effects of dialysis.

The kinetics of 6-[(R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido]-penicillanic acid sodium salt (mezlocillin, Baypen) in renal failure were studied in 23 patients, given 5 g as i.v. infusion. Serum half-lives (t1/2) were around 60 min in normal renal function, 90 to 120 min in patients with a creatinine clearance (Clcr) of 30-60 ml/min, 160 to 190 min at a ClCr of 15 to 30 ml/min and 6 to 14 h in patients with a ClCr below 10 ml/min. There was a linear correlation between serum creatinine and t1/2. Peritoneal dialysis reduced t1/2 to 3 to 4 h and during haemodialysis t1/2 was 1.4 to 2 h with a mean extraction ratio of 0.25. Levels in urine were very high, even in advanced renal failure and the recovery rate was around 60% of dose in normal renal function, but less in renal failure, being only 10-20% at a ClCr of less than 10 ml/min. Levels in peritoneal dialysate fluctuated between 20  and 30 microgram/ml for 4 to 6 h after the infusion, falling slowly but still averaging 10 to 15 microgram/ml after 18 h. Dosage regimens are proposed for various situations in impaired renal function.


Cerebrospinal fluid penetration of mezlocillin.

The passage of 6-[(R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido)-penicillanic acid sodium salt (mezlocillin, Baypen), into the CSF was studied in 9 patients with symptoms of acute meningitis, presumed to be of viral origin. The antibiotic was given as a single 5 g dose i.v. over 30 min. The CSF/serum concentration ratio of mezlocillin showed a variation from 0 to 10.7%. The antibiotic could be effective in the treatment of bacterial meningitis caused by ampicillin-resistant strains of Haemophilus influenzae and by most Enterobacteriaceae, provided these results will be confirmed by a study now in progress. In one patient suffering from meningococcal meningitis this concentration ratio varied between 72% (day 3) and 54% (day 12).


[Mezlocillin concentrations in human bone tissue].

20 patients undergoing extended hip surgery received a single i.v. injection of 2 g 6-[(R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido]-penicillanic acid sodium salt (mezlocillin, Baypen). At different times after the infections but simultaneously 31 samples of blood and bone (spongiosa) were collected for determination of mezlocillin concentrations. On an average, maximum concentrations of approximately 20 microgram/g bone were demonstrated 30 min after injection, these being within the clinico-therapeutical range. The concentration of azlocillin--another acylureido-penicillin--was comparatively higher.


Treatment with azlocillin in complicated urinary tract infections.

20 patients, aged 18-84 years, with complicated urinary tract infections admitted to hospital were treated with 6-[(R)-2-(2-oxo-imidazolidine-1-carboxamido)-2-phenyl-acetamido]-penicillanic acid sodium salt (azlocillin, Securopen) for 5 to 10 days. Isolated bacteria were Pseudomonas aeruginosa (14), Proteus mirabilis (3), Escherichia coli (2) and Klebsiella spp. (1). Serum concentrations and urine recovery were measured on the fifth day of treatment. The mean serum half-life was 1.85 h and the mean value of the urine recovery 47% of the single dose. Tissue concentrations were analyzed in one patient. The samples were taken 3 h after the infusion. The azlocillin concentration of the renal cortex, 60 microgram/g, was six times higher than the corresponding serum concentration. On the 5th day of treatment the urine was sterile in 80% of the patients. In 12 patients (60%) the urine was still sterile when controlled 2-6 months later. Three patients were treated twice and bacteriologically cured after each period. No sign of sensitization was seen. Side-effects were not reported.


Local tolerance of i.m. mezlocillin injection.

To evaluate the intensity of the localized pain caused by intramuscular administration of 6-[(R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido]-penicillanic acid sodium salt (mezlocillin, Baypen), the pain was compared to that caused by ampicillin in a blind test. Ampicillin served as reference because of its extensive use. 50 patients were given 1 g of mezlocillin and 1 g of ampicillin successively, in both gluteal regions. Ampicillin is significantly more painful than mezlocillin. However, substantial pain is produced by both drugs. In a second study, to reduce the pain, 50 patients were given 1 g of mezlocillin twice, one injection was given in a period of 5-7 s, and the other in a 12-15 s period. The slower injection is significantly less painful than the faster one. In conclusion, i.m. administration of mezlocillin is less painful than that of ampicillin, and the slow injection reduces the pain of mezlocillin administration.


[Clinical experience with mezlocillin in infections of mouth and jaw (author's transl)].

The effectiveness and tolerance of the new semisynthetic acylureidopenicillin 6-[(R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido]-penicillanic acid sodium salt (mezlocillin, Baypen) were investigated in 10 patients with bone and soft tissue infections of the mouth and jaw. The antibiotic was given in doses of 2 to 10 g/d. Indications for treatment were osteomyelitis of the jaw and/or phlegmon of the floor of the mouth and abscesses of the soft palate. 9 out of 10 infections could be cured. Due to the short treatment period one case was not assessable. In 6 patients 9 causative aerobic and anaerobic organisms were isolated which were eliminated. The clinical and laboratory control examinations did not reveal any adverse reactions.


Treatment of childhood meningitis with mezlocillin.

30 children suffering from bacterial meningitis and 2 children suffering from septicemia were treated with 6-((R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetmido(-penicillanic acid sodium salt (mezlocillin, Baypen). The daily dose was 250 mg/kg, divided in three portions. Therapy was successful in all patients. Neither signs of toxicity nor side effects of any kind could be found. Mezlocillin concentrations were measured in serum and cerebrospinal fluid (CSF) mainly on days one and six or seven of therapy. Serum concentrations were in the expected range. CSF concentrations depended on the inflammation of the meninges. On the first day of treatment they ranged from 0.5 to 7.2 to 12.0 microgram/ml. After normalisation of CSF no concentrations of mezlocillin were detectable.


[Therapy of pneumonia in children with mezlocillin (author's transl)].

Pneumonia in children requires immediate antibiotic treatment. The identification of the causative organism is very difficult and, therefore, of little importance for therapy. A number of simple examinations, such as BSR, neutrophil count and unsegmented neutrophil count, allow to differentiate between bacterial and viral infections. This method is particularly suitable for assessing an antibiotic treatment retrospectively. These considerations were borne in mind when instituting treatment in 72 babies and children suffering from pneumonia. The following diagnoses were established: 12 cases of whooping cough associated with pneumonia, 4 cases of congenital defects and pneumonia, 1 case of rare Salmonella-induced septicopyaemia and pneumonia, 6 cases of pneumonia and reduced defence against infections and 49 cases of primary pneumonia. Except for one child presenting an insufficiently developed immunity system, all the children were rapidly cured within 5-7 days. The only side effect observed was rash in two cases.


[Experience with the combination mezlocillin/oxacillin in severe bacterial infections in a children's surgical ward (author's transl)].

Severe bacterial infections accompanying conditions during childhood which demand for surgical intervention mostly are caused by Staph. aureus, E. coli, Klebsiella/Aerobacter or Ps. aeruginosa. 20 patients, 10 of them suffering from a putrid peritonitis, showed a good efficacy of Optocillin (Bay 1-1330), a combination of 6-((R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido)-penicillanic acid sodium salt (mezlocillin, Baypen) and 5-methyl-3-phenyl-4-isoxazolylpenicillin (oxacillin, Stapenor), in 85%. Bacteria were eliminated in 79%.


[Clinical studies with a mezlocillin-oxacillin combination in a children's intensive care ward].

25 severely infected children, including 10 premature babies weighing less than 2000 g, were treated with Optocillin, Bay 1-1330 a combination of 6-((R)-2-[methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido)-penicillanic acid sodium salt (mezlocillin, Baypen) and 5-methyl-3-phenyl-4-isoxazolypenicillin (oxacillin, Stapenor). Three children died, but probably only one of them of an (uncomfirmed) infection. Very good therapeutic results were obtained in 10 of the 22 survivors, good results in 11 children, and there was one therapeutic failure. In 9  cases an aminoglycoside antibiotic (mostly amikacin) was given in addition during the critical phase of the disease. The daily dose ranged between 120 and 270 mg/kg bodyweight, corresponding to 80-180 mg mezlocillin and 40-90  mg oxacillin/kg bodyweight. The duration of treatment was between 5 and 38 days. Severe side effects definitely produced by the compound were not observed; a rise in the liver-specific laboratory values of one patient was not definitely attributable to medication. According to this study, the combination mezlocillin-oxacillin can be rated as an antibiotic well effective against a broad range of pathogens and marked by a good tolerance especially in the children's intensive care ward.


[Therapy with an antibiotic combination mezlocillin/oxacillin and clinical experience/Second communication: Children (author's transl)].

Almost all infections of newborn have to be treated with antibiotics before the result of microbiological examination is available since there are either difficulties in identifying the causative organisms(s) or antibiotic treatment must be instituted without delay in view of the patient's life threatening condition. The present paper reports on the therapeutic results obtained in association with the multicentre study of 51 children affected by bacterial infections. 17 (33.0%) patients were newborn up to one week of age. 12 patients were classified as premature babies, some presenting a high degree of immaturity. Treatment with Optocillin (Bay 1-1330), a combination of 6-((R)-2-[3-Methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido)-penicillanic acid sodium salt (mezlocillin, Baypen) and 5-methyl-3-phenyl-4-isoxazolyl-penicillin (oxacillin, Stapenor), proved effective in 90% of the children, although 22 patients had been treated with other antibiotics prior to the use of this compound. An average daily dose of about 225 mg/kg bodyweight was found to be effective and perfectly tolerated even on long-term treatment, as became evident from two cases of extremely premature babies effected by neonatal septicaemia (duration of treatment 38 and 34 days, respectively). The combination showed a perfect local tolerance, exanthema appearing only in one infant.


Changes in metabolic activities of Fusarium oxysporum f. fabae and Rhizoctonia solani in response to Dithan A-40 fungicide.

The effect of different concentrations of Dithan A-40 fungicide on the metabolic activities of the wilt fungus Fusarium oxysporum f. fabae and the root rot agent Rhizoctonia solani was studied. All toxicant concentrations reduced energy generation, total phosphorus and nitrogen content of both fungi. In addition, the toxicant caused a shift in free amino acids pool. As a result of these changes, the mycelium dry weight of both fungi was greatly reduced. R. solani was more sensitive to the toxic effect of Dithan A-40 than F. oxysporum.


[Influence of moisture on the nitrification of two soils of central Bohemia (Czechoslovakia) (author's transl)].

The production of N-NO3 as a function of initial moisture (pFinit), incubation moisture (pFinc) and time of incubation was investigated in two soils in Prague area. The results show the existence of linear relationships between the production of N-NO3 (expressed in logarithm of p.p.m.) and pFinit, pFinc, time of incubation (expressed in logarithm of number of days).


Comparative studies with regard to the influence of carbon and nitrogen ratio on sporulation in Fusarium oxysporum and Fusarium moniliforme v. subglutinans.

Carbon/nitrogen ratio as a factor for sporulation, expressed in terms of magnitude of population variation of macroconidia and microconidia in the cultures of Eusarium oxysporum Schlecht ex. Fr., Fusarium moniliforme v. subglutinans Wr. and Rg., and of chlamydospores (only in Fusarium oxysporum) was investigated. It has been found that the amount of carbon source shapes the course of macro- and micro. conidial production in a linear fashion, being enhanced parallel to the increase in its amount-Nitrogen level, limiting proliferation and effectively diminishing the macro- and micro-conidial population, varies for the two species, namely Fusarium oxysporum and Fusarium moniliforme v-subglutinans. For chlamydomspore production, higher carbon and still higher nitrogen concentration favours profuse proliferation in case of Fusarium oxysporum.


The influence of mutagenic chemicals on lactic acid fermentation by Lactobacillus bulgaricus AU.

Lactic acid fermentation from molasses by L. bulgaricus AU in the presence of caffeine and ammonium fluoride was studied. It was found that caffeine had a stimulating effect only at lower concentration (0.001%-0.002%), while at high concentration the acid-producing activity of the bacteria was considerably depressed. Ammonium fluoride, at a concentration of 0.001%-0.005%, had a stimulating effect on the activity of the bacteria, maximum activity was observed at a concentration of 0.003%.


Correlation between macrophage intracellular electrical potentials and malignant melanoma growth in a murine model.

Peritoneal macrophages were collected from mice at varying periods after transplantation of an allogeneic malignant melanoma in the hind limb. The intracellular electrical potentials of these macrophages were measured and a correlation was found to exist between tumor growth measured by size and pathological examination, and the development of large negative intracellular potentials. We propose that this change in intracellular potential is correlated with changes in the immune system and may be triggered by membrane permeability changes possibly in response to calcium ions.


A potent inhibitor of cell proliferation in "middle molecules" isolated from the urine of uremic patients.

A potent inhibitor of cell proliferation was found in the urine of a patient with chronic renal failure. This substance included in "middle molecules" (MM) fraction, was obtained by chromatography. This factor was shown to inhibit noticeably the proliferation of various cells : lymphocytes stimulated by allogeneic cells, monolayer cell lines of normal or tumorous origin and leukemic cell lines derived from acute lymphoblastic leukemia. This effect was reversible and thus could not be related to a direct, rapid cytotoxic effect of MM. Such substances could play an important part in uremic symptoms, such as immunodeficiency, anemia, thrombopenia, gastrointestinal or skin manifestations.


Lymphokine-induced changes in macrophage transmembrane electrical potentials.

Intracellular electrical potentials have been measured in nonactivated and immunologically-activated macrophages obtained from the peritoneal cavities of mice. Normal macrophage potentials were established and found to become significantly more electronegative after in vitro exposure for 10 minutes to a lymphokine-containing supernatant which induced macrophage activation. This approach may reflect very early concomitants of such activation and is also useful in the study of other immunologic systems.


Some causes of blindness seen at Noor Eye Institute, Afghanistan.

This is a survey of the causes of blindness recorded by three students of The City University during a six-week stay at NOOR Eye Institute in Kabul, Afghanistan. The nature of the sample used makes it unsuitable for direct comparison with surveys from other countries. Nevertheless, broad conclusions can be drawn. Severe eye disease is a considerable problem in Afghanistan, in common with other developing countries. The survey included all new patients seen in the outpatient clinic, blind to the extent of being unable to count fingers at more than three metres, in one eye or both. The sample of patients was examined by an ophthalmologist working with our team. A diagnosis was made and various social and demographic questions were asked with the help of an interpreter. In all, 473 patients were examined and included in our survey sample, a very high proportion of the new patients attending the clinic (40.46 per cent). The main causes of blindness were found to be cataract (31.12 per cent), corneal scarring (19.8 per cent), chorioretinal degenerations (6.79 per cent), glaucoma (6.65 per cent) and aphakia (5.52 per cent). Of the major causes of blindness, about forty per cent of the cases were considered remediable, and about thirty per cent could have been prevented. An outstanding difference between the causes of blindness in developed nations and those seen in Afghanistan was the amount of blindness caused by infection, especially in the younger age groups (up to 30 years). Cataract is a major cause of blindness in the older age groups of both societies. People suffering from ocular disease in Afghanistan wait until their sight is badly impaired or lost completely before seeking treatment. This results from a lack of knowledge of what could be done to conserve sight, the irreversible nature of many eye diseases, the distances involved in travelling to the clinic, and even a lack of knowledge of its existence.


Seasonal acclimatisation in the hen.

1. The oxygen consumption and body temperature of White Leghorn x Rhode Island Red laying hens, which were being kept in a semi-natural climate, were periodically measured for short periods at ambient temperatures between 2 and 32 degrees C in a suitable chamber. 2. A seasonal shift was observed in lower critical temperature (Tc) and thermoneutral oxygen consumption. The slope of the relationship between oxygen consumption and air temperature at less than Tc was not discernibly affected by season. 3. The seasonal shifts correlated best with mean daily outside temperature Ta) during the experimental periods; a seasonal decrease of 10 degrees C in Ta was associated with a decrease of 8.5 degrees C in Tc and a 12% increase in thermoneutral oxygen consumption. 4. The capacity to maintain body temperature stability at extremes of environmental temperature was improved by acclimatisation to heat.


Effects of food intake regulation on the energy metabolism of hens and cockerels of a layer strain.

1. Indirect calorimetry was used to study the effects on the energy metabolism of cockerels and hens after peak lay of a hybrid layer strain of regulating food or energy intake to 80% of ad libitum for an extended period of time. 2. Regulation of food intake decreased fasting and fed heat productions per bird and per unit metabolic body weight (kg0.75). 3. Maintenance energy requirements (per kg0.75) of both sexes were reduced about 20% by regulation; maintenance requirements of cockerels were about 30% lower per kg0.75 than hens on the same treatments. 4. Reducing the intake of all nutrients by 20% had the same effect on the energy metabolism of cockerels as restricting energy alone by the same amount. 5. Gross efficiency of egg production, in terms of both mass and energy, was higher when intake was regulated. During weeks 21 to 52 of restriction, daily egg mass output was significantly higher in hens fed ad libitum. 6. After 52 weeks of food regulation, restricted hens weighed 20% less and their total carcass energy was 36% less than hens fed ad libitum; similar effects were found in the cockerels.


Effects of water temperature on short-term water intake and medullary neuronal responses in the hen.

1. The daily water intake of adult Brown Leghorn hens was measured at water temperatures of 0, 10, 20, 30, 40 and 45 degrees C. The electrical activity of neurones in the nucleus ventro-lateralis anterior solitarii (NVLAS) of the medulla was also measured when the mouth was stimulated with water at the same temperatures. 2. There was a significant decrease in water intake when the temperature of the water was 10 or 45 degrees C. 3. There was an increase in activity of some of the NVLAS neurones with decreasing water temperature and this activity was in response to specific "cold" receptors. 4. Some NVLAS neurones were identified which showed an increase in activity at temperatures of 45 degrees C and more; it was concluded that these represented responses to thermal pain. 5. It is concluded that the effects of temperature on water intake may in part be explained by the behaviour of the oral receptors.


Water intake of laying hens after exposure to a warm climate.

1. The water intake of laying hens was measured after exposure for 4 h to an ambient temperature (TA) of 35 or 40 degrees C. 2. Water intake increased continuously over 1 h but did not exceed a mean value of 80% of water lost by evaporation. There was considerable variation between birds.


Trimethylamine and egg taint.

1. The sensory qualities of "crabby", "fishy" tainted eggs are closely similar to those of an aqueous solution of trimethylamine (1 x 10(-2) microgram/g). 2. The odour threshold of trimethylamine in water is 5 +/- 1 x 10(-4) microgram/g. 3. The taint occurs in eggs which contain more than 0.8 microgram/g trimethylamine. The reasons why this concentration is higher than that in water are discussed.


Early body-weight selection of broiler breeder males in relation to reproductive and growth performance of their offspring.

1. The effectiveness of selection of broiler breeder males for body weight at 3 weeks of age on later growth, semen quality and performance of progeny has been tested. 2. Correlation between 3- and 20-week body weights in the breeders was poor but significant. 3. On the basis of 3-week body weight males were divided into heavy birds (mean + 0.5 standard deviation) and all birds. 4. Semen quality was not different between the two groups, but some selection for semen quality was practised within groups. 5. There were no differences in fertility and hatchability of eggs produced from hens artificially inseminated with diluted, stored semen from both groups. 6. There was a small, positive, but non-significant, effect of selection of breeder males on body weight of progeny at 6 weeks of age.


Further studies with a sex-linked lethal gene in the fowl.

1. The ladykiller (lk) gene has been located on the sex chromosome of the fowl. It is 4.2 map units distant from the feathering rate locus (k) and 11.2 map units distant from the dwarf (dw) locus. 2. Most of the embryonic mortalities associated with this gene occurred between 3 and 4 d of incubation.


The relationship of several genes suppressing plumage colour with body weight, food intake and feather loss of laying hens.

1. Growth rate, egg number, egg and adult body weight, plumage condition, food intake and efficiency of laying hens were compared in birds differing in plumage colour genotype at five loci (C, I, S, Ig, B). 2. Laying rates of the different genotypes were not different. 3. Growth rate and adult body weight of females were significantly less in loci C and I white females as compared with the corresponding coloured females; there was a similar tendency in adult weight for the other loci, and in egg weight. Differences in body weight were more marked in birds kept in cages than in those kept on the floor.  4. For all loci the genotype suppressing colour was associated with less plumage deterioration, this being highly significant for loci C and S. 5. Food intake, adjusted for body weight and egg production, was less in locus I white hens, in agreement with previous data concerning the C locus. 6. Pleiotropic effects associated with colour were suggested and their consequences on efficiency of egg production are discussed.


An electrophysiological quantitation of the cubital tunnel syndrome.

Four hundred and ninety-two ulnar nerves were studied in 237 patients with cubital tunnel syndrome and 233 subjects without clinically apparent ulnar neuropathy. Terminal motor and sensory latencies, motor and sensory conduction velocities, sensory action potential amplitudes and dispersons, and needle electromyography were analysed by a 0 to 10 rating--the EMG score. The patients were clinically graded from 1 (least severe) to 4 (most severe). The median EMG scores of these were 0.9/10 (N = 70); 1.6/10 (N = 81); 4.7/10 (N = 46); and 7.1/10 (N = 56), respectively. The median EMG score for the controls was 0.6/10 (N = 239) and 14.7% had asymptomatic ulnar entrapments. This incidence reached 30% in controls who were 60 years or older. In 25 other controls, the above sulcus sensory action potential was analysed. Its mean dispersion was 4.9 +/- 1.0 s-3. This measurement was significantly prolonged in 84.6% of 13 studies in which the EMG score was 0/10. These patients had either sensory symptoms only or an additional mild sensory deficit. Comparison of the EMG score obtained with sequential studies would allow one to objectively evaluate improvement or deterioration of the clinical state, giving a rational means of determining the need for surgery.


Evaluation of cerebral blood flow in arteriovenous malformations by the xenon 133 inhalation method.

Fifteen regional cerebral blood flow studies (rCBF) were conducted on 14 patients with arteriovenous malformations (AVM). Only one patient was studied at the time of a hemorrhage. None of the patients were operated upon. All patients had angiographically demonstrated lesions. All the CT scans performed demonstrated the lesions. rCBF was increased in the involved hemisphere compared to the non-involved hemisphere and the difference was greater when the malformations were superficial. There was a higher incidence of high flow regions in the involved hemisphere of patients with AVMs compared to our normal control group and patients with subarachnoid hemorrhage (SAH) from aneurysms. Flow rates within the involved hemisphere demonstrated abnormal distributions and greater variability than normals. Some peaks previously demonstrated in the intra-arterial flow studies are not apparent using the inhalation method. This non-invasive technique has a potential to provide additional useful information on the natural history of this disorder.


Methylmercury poisoning in Canadian Indians--the elusive diagnosis.

A World Health Organization Expert Committee has concluded that symptoms of methylmercury poisoning may appear at blood concentrations of 200-500 ng Hg/ml. Blood levels in this range have been found in several Indian and Inuit communities in Canada. The syndrome of severe methylmercury poisoning (Hunter-Russell syndrome) is well described. However, diagnosis of less severe cases is difficult. This paper review the present situation in Canada. The problems of diagnosis currently being encountered are discussed and are illustrated by the case of an individual who had one of the highest blood concentrations (551 ng Hg/ml) ever described in fish-eating populations outside of the outbreaks in Minamata and Niigata in Japan. Although mercury concentrations in brain were estimated to have been in the "symptomatic" range at least once in the two years prior to his death, neurohistological examination was normal. Detailed examination by two teams of neurologists revealed effects that may be associated with methylmercury poisoning but a definitive diagnosis remained elusive.


Tissue albumin and water content in the early stages of vasogenic brain edema formation.

Brain edema (BE) research lacks quantitative regional methods. We modified the method of Pappius and McCann (1969), who used radioactive iodinated I125 serum albumin (RISA) as a label for vasogenic BE fluid. To correct for intravascular plasma volume we used Cr51 labelled red blood cells and calculated equivalent extravascular plasma volume (EVPV). The modified RISA method was compared with a standard method for measuring increased tissue water, i.e. the change in wet:dry weights. Anesthetized rabbits were subjected to unilateral cortical freeze injury and sacrificed three hours later. The lesion corresponding to the area of blood brain barrier (BBB) breakdown was delineated by Evan's blue staining. That area and the two adjacent poles were sectioned. The contralateral hemisphere was used as the control. Good agreement was found between the two methods. By the modified RISA method, 4.08% of the wet weight of the injured hemisphere was EVPV while the decrease in dry weight corresponded to 5.56% edema. In the freeze lesion, however, where BBB breakdown occurred extravascular (EV) protein exceeded EV water, while adjacent to the lesion in the area with intact BBB, EV water exceeded EV protein. This suggests that the diffusion rate for water through brain tissue exceeds that of albumin in the early stages of vasogenic BE formation. We conclude that the modified RISA method is a satisfactory measure of regional vasogenic BE in acute animal experiments.


Examining survival data.

This brief review presents a nonmathematical description of the methods used to describe the outcome of patient groups when the time to an event such as death or disease recurrence is of interest. Calculation of the product-limit, actuarial and relative survival curves is described and the underlying principles are explained. Two statistical tests frequently used in comparing survival curves (the generalized Wilcoxon and the log-rank or Mantel test) are described and an example of their use is given.


Drug prescribing for the elderly in Saskatchewan during 1976.

Over 11% of Saskatchewan's population is 65 years of age or older. To study patterns of prescribing for the elderly a computer file of more than 3.6 million prescriptions filled in Saskatchewan in 1976 was reviewed. The elderly population (aged 65 years or more) that year was 102 070, and 77.3% received at least one prescription drug listed in the Saskatchewan Formulary. In comparison with a "middle-aged" group (persons between the ages of 35 and 54 years) the elderly showed moderately higher average numbers of prescriptions filled and drugs used per person that year. However, the prescription of antihypertensive agents and diuretics increased dramatically with age, and barbiturates, in view of their potential toxicity, appeared to be overprescribed for the elderly.


Case report: equine colic associated with sand impaction of the large colon.

In most cases the outcome of impaction of the colon is favourable and with mild purging and lubrication, the mass can be broken down and the horse returned to normal health. It is important to reevaluate impaction cases which neither deteriorate or improve in a 24 hour period and to perform a paracentesis to evaluate the etiology and prognosis. A decision of euthanasia or surgical correction should be made as early as possible. Though initially stable, vital signs will deteriorate rapidly without warning. A critical evaluation of the client's farm management can prevent many impaction colics and the service practitioners provide is not complete unless the necessary changes are made.


Case report: fibrocartilaginous embolic myelopathy in a dog.

This case report gives details concerning fibrocartilaginous arterial embolism leading to an ischemic myelopathy in a dog. Using various histochemical techniques, the material occluding the vessels was found to be similar to that reported for fibrocartilaginous ground substance of the nucleus pulposus. The pathway by which this material enters the arterial vasculature of the spinal cord is not known.


[Dermatophilus congolensis infection in a dairy cow in Quebec (author's transl)].

Dermatophilus congolensis infection in a dairy cow in QuebecA case of infection by Dermatophilus congonlensis is described for the first time in the Province of Quebec, in a dairy cow. Dermatophilosis was diagnosed in the area of Saint-Hyacinthe in October 1978. It was not possible to find the source of the infection. The isolation of the microorganism was successful and the three techniques used are described. The death of the animal was attributed to complications following invasion by a secondary pathogen. All the other animals in the herd were free from clinical signs of infection. Moreover, no other case of infection by D. congolensis was found in other animal species or humans in the area.


Cytocidal effect of rifamycin derivatives on ascites tumor cells: studies with [125I]iododeoxyuridine.

The cytotoxicity of 2 rifamycin derivatives, rifazone-82 and rifampicin, on mouse ascites cells was studied, using the [125I]iododeoxyuridine (IUDR) method of labeling the tumor cells. This technique allows a distinction to be made between a cytocidal and cytostatic effect. The 2 drugs exerted a cytocidal effect against 2 non-leukemic cell lines, but had no effect against 3 leukemic lines.


The effect of time on the incidence of carcinomas obtained by the implantation of paraffin wax pellets into mouse bladder.

Paraffin wax pellets were implanted surgically into the mouse bladder lumen and animals killed after 40-50, 70-80 and 100-110 weeks. The number and malignancy of bladder carcinomas increased considerably with the time after implantation. Pellets containing 1-phenylazo-2-naphthol significantly increased at each time the incidence of bladder carcinomas relative to that observed with paraffin wax. Pellets containing N-2-fluorenylacetamide or its metabolites gave very similar incidences or carcinomas whether or not they were carcinogenic by oral or parenteral administration.


Identification of transformed rat liver epithelial cells in culture.

The growth potential of normal and transformed rat liver epithelial cells in culture was investigated under various experimental conditions. The control cells failed to form colonies over a base layer of living parent cells, whereas transformed cells formed distinct colonies. Contact with living normal liver cells is required for the growth inhibition of the untransformed cells. Malignant cells capable of producing hepatocellular carcinomas in vivo exhibited signs of invasion in vitro. Altered properties of hepatic cells during the early stages of transformation can be assessed by plating such cell population over a living base layer of normal liver cells.


Microsomal biphenyl 2-hydroxylation in the presence of some carcinogenic alkylaryltriazenes.

A hepatic mixed-function oxygenase system gave enhanced biphenyl 2-hydroxylation in the presence of known carcinogens in vitro and was therefore suggested as a convenient short-term assay for the detection of potentially dangerous compounds. However, the test did not yield the expected increase of biphenyl 2-hydroxylation with any of 8 selected carcinogenic triazenes. The new data are discussed in relation to the original supposition and to the hydroxylation pattern of various triazene substrates in vivo.


Androgen, estrogen and progesterone receptors of the R3327H Copenhagen rat prostatic tumor.

Sucrose density gradient analysis of the R3327H tumor cytosol demonstrated the presence of both androgen and estrogen binding proteins. Competitive binding analysis with 17 beta-estradiol, 5 alpha-dihydrotestosterone, R1881, cyproterone acetate, and cortisol was consistent with the presence of 2 different binding sites for androgens and estrogens. Scatchard binding analysis was performed in the dorsal-lateral prostate as well as the R3327H tumor from normal Copenhagen rats. High affinity receptors for androgen and estrogen but not progesterone were found. However, in R3327H tumors relapsing following castration, the presence of high affinity receptors for progesterone were readily detectable.


Carcinogenicity of di(N-nitroso)-perhydropyrimidine (DNPP) after intraperitoneal application to rats.

Male Sprague-Dawley rats received 30 mg/kg body wt. (group I) or 15 mg/kg (group II) di(N-nitroso)-perhydropyrimidine (DNPP) by the intraperitoneal route once a week for life. The DNPP treatment significantly reduced the life expectancy in both groups. DNPP induced tumors of the esophagus (27% incidence in group I, 33% in group II) and possibly also of the liver (4% incidence in group I, 8% in group II).


Amino acid conferred protection against melphalan interference with melphalan therapy by L-leucine, a competitive substrate for transport.

Melphalan uptake by L1210 leukemia cells obtained from tumor bearing mice is reduced to one-third of control by physiological concentrations of L-leucine. Kinetic analysis revealed that melphalan and leucine compete for transport carrier sites. Administration of leucine with optimal therapeutic doses of melphalan to tumor bearing mice negated the efficacy of the drug.


Effects of morphine on left ventricular dimensions and function in patients with previous myocardial infarction.

To assess the effects of morphine sulfate on left ventricular function and dimensions we administered 15 mg of this agent to 11 stable patients with previous transmural myocardial infarction. All studies were carried out in the supine position. Before morphine administration an echocardiogram was obtained, and this procedure was repeated at 15, 30, 60, 120, and 240 min after morphine. Heart rate decreased from a control value of 69 +/- 4 to 62 +/- 5 beats/min 2 h after morphine (p less than 0.01, analysis of variance); this slower heart rate persisted for 4 h after morphine. Serial measurements of blood pressure, echocardiographic ejection fraction, percent of fractional shortening, and mean normalized velocity of circumferential fiber shortening also showed no significant alterations after morphine. We conclude that in stable patients with chronic ischemic heart disease studied in the supine position, 1) morphine exerts no effect on left ventricular dimensions, an observation which does not support the concept that this agent acts in humans by producing a 'pharmacologic phlebotomy'; and 2) morphine does not alter left ventricular function at rest. Whether different results will be found in patients with increased sympathetic activity, such as occurs in the setting of an acute myocardial infarction or during an episode of acute pulmonary edema, remains to be investigated.


Quantification of myocardial ischemia and infarction with 201thallium scintigraphy.

A quantitative method for the analysis of 201thallium myocardial scintigrams, developed in an experimental infarcted dog heart model, has been compared with two nonquantitative methods for interpretation of stress myocardial scintigrams in two groups of patients studied with coronary angiography: 11 with normal coronary arteries and 14 with coronary artery disease. Three independent observers interpreted scintigrams which were 1) not computer processed; 2) corrected for background activity in lungs and chest wall; and 3) processed by a computer method which uses a uniform threshold of counts determined from the dog model to define perfusion defects. Interobserver variability as well as sensitivity and specificity of detecting coronary disease were examined. In patients with coronary artery disease interobserver variability was improved by using the computer technique: observers agreed as to the existence of a perfusion defect in 93% of the scintigrams as compared to 55% and 81% for the unprocessed and background-subtracted images respectively. No false positive indications of coronary disease were obtained by any of the three techniques. Use of the computer method did not improve the sensitivity of detecting coronary disease, however--71% compared to 64% for unprocessed images and 79% for background-substracted images. The advantages of this quantitative computer method are increased consistency of interpretation and lack of false positive diagnoses of coronary disease. An improved sensitivity of detection may be gained by varying thallium count thresholds according to anatomic location in the heart.


The prognosis of anterior myocardial infarction revisited: a community-wide study.

A study was carried out in metropolitan Baltimore in which the short- and long-term prognosis of 655 patients with anterior myocardial infarction (MI) was compared with that of 520 patients with inferior/posterior MI. The study was performed on a community-wide basis in two time periods before the clinical introduction and widespread use of beta-blockade therapy. After simultaneous adjustment for several socio-demographic and clinical variables, the in-hospital case fatality rate was greater for patients with anterior (27.5%) than for those with inferior/posterior MI (22.9%) (p less than 0.05). Similarly, for those discharged alive from the hospital and followed up for as long as 10 years, patients with infarction of the anterior wall had a significantly poorer long-term survival than that of patients with infarction of the inferior/posterior wall (p less than 0.05). These results suggest that closer surveillance should be directed at patients with anterior myocardial infarction, both during the acute phase and after hospital discharge.


Natural history and pathophysiology of arteriovenous malformations.

AVMs are congenital malformations of central nervous system blood vessels. The majority of lesions become symptomatic prior to age 40. Hemorrhage and epilepsy are the most frequent presenting symptoms. Disabling headache, transient, progressive, and permanent neurological deficit, heart failure, hydrocephalus, macrocephaly, and intellectual impairment may also occur. Long-term mortality is approximately 18%, and morbidity leading to disability occurs in 30% of patients. Aggressive management when possible is indicated.


Microsurgical approach to orbital tumors.

Neurodiagnostic techniques now permit precise localization of intraorbital tumors relative to the optic nerve and intracranial compartment. With such localization, surgical procedures may be planned along more anatomical lines. Those circumscribed tumors may be planned along more anatomical lines. Those circumscribed tumors lateral, superior, and inferior to the optic nerve may well be approached by a lateral orbitotomy using microdissecting techniques. Those tumors medial to the optic nerve deep in the apex and those with possible intracranial extension should be approached transfrontally. In both surgical approaches, precise anatomical knowledge and microdissecting techniques are needed.


Multiple bleeds in haemophilia A.

One hundred and eighty-one bleeding episodes involving two sites simultaneously were noted during a survey of 4935 bleeding episodes, an incidence of 3.7%. Elbows, knees and ankles were the commonest sites involved in double bleeds, while the thigh, upper arm and elbow were the commonest sites involved in the double bleeds needing most transfusions. The overall transfusion requirements were less than for single bleeds. The frequency of multiple bleeds correlated significantly with the overall bleeding frequency, but not with the number of days under observation.


The use of Dispholidus typus venom in elucidating the cause of a low prothrombin index.

A simple and rapid in vitro test was devised to detect the presence of the prothrombin precursor molecule circulating in the plasma of 55 out of 56 patients receiving oral vitamin K antagonists and in 4 out of 5 patients with obstructive jaundice, using either Dispholidus typus or Echis carinatus venoms. The absence of the molecule from the plasma of jaundiced patients was suggestive of hepatocellular rather than obstructive jaundice. The test is based on the clotting of aluminium hydroxide gel adsorbed plasma by the venoms. It is more sensitive and easier to perform than measuring antigenic prothrombin levels.


The low mean cell volume in routine haematology.

This study establishes the prevalence and distribution of an MCV less than 80 fl by review of the Coulter indices of 7887 samples. Retrospectively, 146 patients are studied and the relationships between the low MCV, film microscopy, iron status, and clinical diagnosis assessed. Plasma ferritin measured radioisotopically in 100 patients is compared to serum iron and iron binding capacity values and some of the problems of assessing iron stores are discussed. The discriminant formulae devised to differentiate between the microcytosis of thalassaemia trait and that or iron deficiency are found to be of limited value when applied to the red cell indices of a hospital population.


A laboratory evaluation of Compur M 1000 Miniphotometer and the Compur M 11OO minicentrifuge.

The Compur Miniphotometer and Minicentrifuge are portable instruments designed for rapid estimation of haemoglobin, red cell count and haematocrit on capillary blood. Haemoglobin and haematocrit results correlate well with reference methods. Accurate red cell counts are only obtained when the sample MCV is within the range 83--103 fl, thus restricting the application of this method. The Compur M 1000 Miniphotometer and Compur M 1100 Minicentrifuge have been designed as portable instruments to permit rapid estimations of haemoglobin concentration, red cell count and haematocrit by the non-laboratory worker.


A computer system for haematology. I. System design and data acquisition.

The configuration for a comprehensive computer system in haematology is given including the hardware and a general description of the software. Details are provided of the application programs and the methods used for test data acquisition and verification.


A computer system for haematology. 2. Handling, storing and transmitting of data.

Using a system described elsewhere, a suite of programs to collect, verify, store and retrieve patient and laboratory data were created. The way in which this is carried out and the lessons to be learnt for the future are described.


Effect of cardiac pacing on ECG pattern in patients with myocardial infarction.

The ECG tracings in patients with recent and past myocardial infarction and permanent or temporary artificial pacing were analysed. The observations were made in 12 patients (8 with recent infarction and 4 with a past one). Anterior wall infarction was diagnosed in 4 subjects, anteroseptal in 3, inferior wall infarction was recognized in 4, and posterior wall infarction in one patient. In the stimulated complexes of persons with anterior and anteroseptal infarctions there appears a pathological Q deflection with ST segment elevation in leads I, aVL, V5. In the infarct of the inferior wall the ST segment elevation occurs in leads II, III, aVF without the presence of Q deflection. No differences were shown in the morphology of stimulated complexes in persons with recent and past myocardial infarction.


The use of the first derivative of apexcardiogram for determining some systolic and diastolic time intervals in ischaemic heart disease.

In 50 normal subjects and in 63 patients with ischaemic heart disease, the left ventricular apexcardiogram with its first derivative (dA/dt), electrocardiogram and phonocardiogram were recorded simultaneously. This allowed to measure four systolic and diastolic time intervals using the first derivative of apexcardiogram. In patients with ischaemic heart disease in increase of the interval R to peak dA/dt, systolic upstoke time of dA/dt, and early apexcardiographic relaxation time (the time from the onset of the aortic component of the second heart sound in the phonocardiogram to the negative peak of the dA/dt) was established. It is concluded that dA/dt is a suitable technique for more exact non-invasive determination of some time intervals than conventional apexcardiogram in ischaemic heart disease.


Diagnosis of intraventricular conduction disturbances with the aid of orthogonal electrocardiography.

In 104 patients with various type of bundle branch block the three most known systems of orthogonal ECG leads (Frank; McFee-Parungao; Beswick-Jordan) were compared with 12 routine leads. All three orthogonal systems proved to possess good informative value, not inferior to that obtained by routine ECG. Examinations of 123 patients suffering from ischaemic heart disease with signs of diffuse myocardial changes with the aid of the Frank's system of orthogonal ECG confirmed the high informative value of the system in the detection of local disturbances of intraventricular conduction. Criteria of evaluation of orthogonal ECG tracings in the detection of left anterior, right, and left bundle branch block were elaborated. The high informative value and limited number of leads recommend the orthogonal ECG system for wide practical use.


Functional states of various heart segments during protracted hypokinesis in patients with tuberculosis of bones and joints.

By the method of acceleration cinecardiography the effect of hypokinesis on the contracting activity of the left and right heart was studied in 46 patients with tuberculous affections of bones and joints, confined to strict bed rest. As controls here served 40 patients with unrestricted mobility and 33 healthy subjects. It was found that the restriction of mobility and compulsory horizontal body positions of the patients contributed to a rearrangement of the phasic structure of the cardiac cycle. During the initial phase of immobilization the unusual conditions of circulation and the specific intoxication enhanced the cardiac contracting activity. From the third month of bed rest onwards, myocardial hypofunction set on in both ventricles and the left atrium. Accelerated heart rate during the hypokinetic period attested to a low efficacy of cardiac work. Disturbances of heart dynamics did not cease even three month of bed rest onwards, myocardial hypofunction set on in both ventricles and the completely controlled.


Shifts in external iliac venous pressure under local and general anaesthesia. Their impact on the tactics of venous thrombectomy in iliofemoral thrombosis.

The authors measured the venous pressure in the iliofemoral segment in 12 patients in good general conditions, with a normal patency of the inferior caval vein, profound pelvic veins, and lower limb veins. Examination was performed with Claudy manometer. The resting venous pressure in the external iliac vein was 40--75 mm H2O = 3--5.5 mmHg. During Valsalva's manoeuvre the patients achieved an overpressure 250--1 100 mm H2O = 18--81 mmHg for 20 s. After induction of general anaesthesia and intubation, the anaesthesiologist produced an overpressure of 50 cm H2O in the patient's respiratory circuit for 20 s, but the venous pressure rose only to 90--175 mm H2O = 7-- mmHg. This rise is lesser with a high statistical significance than the overpressure produced in the Valsalva's manoeuvre. In the light of these results the authors discuss the tactics of venous thrombectomy. As a safe prevention of peroperative uplmonary embolism they regard either Valsalva's manoeuvre, carried out under local anaesthesia, or a tourniquet fixation of the clot head during the surgical intervention under general anaesthesia. The anaesthesiologist cannot prevent embolism by restriction of the venous return by producing an overpressure in the respiratory circuit of a patient under general anaesthesia.


The effects of cold and restraint on the glucose level, bilirubin level and the production of stressed ulcers in rats.

The effect of restraint and cold on the glucose level, bilirubin level and production of stress ulcers were studied in white, adult male, Sprague-Dawley rats of the Holtzman strain. In addition, the role of mucin in reducing incidence of gastric lesions produced by stress was investigated. The highest incidence and depth of ulceration was produced by the stress factor combination of fasting for 24 h followed by physical restraint and cold exposure for 2 h. Under these conditions only one of the experimental rats died throughout the experimentation period. Fasting alone did not produce ulceration. The stress factor combination of fasting for 24 h, followed by physical restraint and cold exposure for 2 h resulted in an increase in glucose level. Increase in bilirubin level was highest in the stress factor combination of fasting, physical restraint and cold exposure acting synergistically. It can thus be hypothesized that the higher the bilirubin level the greater is the rate of red blood cell destruction. Introduction of 2 ml mucin directly into the stomach of the stressed rats reduced ulceration incidence by more than 50%.


Insect sex chromosomes. IV DNA replication in the chromosomes of Gryllotalpa fossor.

In Gryllotalpa the cell cycle duration in the hepatic caecae in vivo is about 12.5 h and of various phases are, G2 + P about 10 h, S about 2.5--3.5 h, and G1 appears negligible or absent. These estimates of the cell cycle are the only ones available in Gryllotalpidae. In the female Gryllotalpa, as in mammals, there is asynchronous DNA replication between the two euchromatic arms of the two X chromosomes. The other arm is constitutively heterochromatized and as expected is late replicating. Thus, a regulatory mechanism for dosage compensation by X chromosome inactivation appears to be operating in Gryllotalpa. This we believe, is the first cytogenetic demonstration of such a mechanism outside mammals.


The ultrastructural localization of acetylcholinesterase activity in the scolex of the cysticercus of Hydatigera taeniaeformis.

Acetylcholinesterase activity in the scolex of the cysticercus of Hydatigera taeniaeformis was studied by means of electron microscopy. Acetylthiocholine iodide was used as the substrate and 10(-4) M of eserine used to inhibit acetylcholinesterase activity. The sites of activity were visualized within the microtriches of the distal cytoplasm, muscle bundles, cytoplasm, vesiculated bodies, flame cells and hooks of the rostellar region. Enzyme activity was also localized within the nuclear membrane, neuroplasm, and membranes of nerve cells.


Failure of tumor promoter 12-O-tetradecanoylphorbol-13-acetate to displace estradiol from its specific receptor.

A permanent cell line derived from rat endometrium which contains a specific, low capacity, high affinity estrogen binding protein in cytosol and nuclear fractions (estrogen receptor) is available. Extracts of cells from this line did not appreciably bind the tumor promoter, 12-0-tetradecanoylphorbol-13-acetate. The result suggests that the action of the promoter does not involve translocation to the cell nucleus via binding to the specific estrogen receptor.


Radioimmunoassay of ovine and bovine serum progesterone without extraction and chromatography.

A procedure for the radioimmunoassay of ovine and bovine serum progesterone is described which does not require extraction and chromatography. Serum samples are assayed directly, and a highly specific antiserum that was prepared in rabbits against 11 alpha-hydroxyprogesterone conjugated to bovine serum albumin is used. Interference from serum binding proteins is alleviated by use of a phosphate buffer containing 5% BSA and separation of bound and free tritiated progesterone by a double antibody procedure. Serum samples are assayed in a mini-vial, the bound fraction (double antibody precipitate) is mixed with scintillation solution and the radioactivity is counted in the same vial. The assay procedure is sensitive (10 pg, 100 pg/ml) and has acceptable accuracy and precision. Because there is no extraction or chromatography, serum progesterone is not lost. Most important, the procedure is specific for progesterone and measures serum progesterone concentrations in the ewe and cow which are comparable with those obtained with conventional assay techniques. The progesterone assay described herein provides a rapid, economical procedure that can facilitate the study of ovarian cyclicity and aid in the early diagnosis of pregnancy.


Alteration of tissue zinc distribution and biochemical analysis of serum following pinealectomy in the rat.

Eight weeks following pinealectomy in adult male Wistar rats, zinc levels of various tissues were found to be significantly altered: zinc in thoracic aorta was significantly increased, and in serum, pituitary, adrenal, heart, lung, and body hair, it was decreased. Serum biochemical analysis indicated that there was a significant elevation of cholesterol, alkaline phosphatase, sodium, urea, and creatinine in serum from pinealectomised rats. Liver, spleen, and thymus weights were lower following pinealectomy, although hearts were increased. The effects of pinealectomy on zinc levels in serum and tissues and on serum cholesterol and alkaline phosphatase may be related to its effects on vascular reactivity and liver fibrosis.


[Partial purification of hog thyroid peroxidase using detergent treatment and its spectral changes induced by hydrogen peroxide (author's transl)].

Thyroid peroxidase (TPO) was partially purified from hog thyroid microsomes after solubilization by means of deoxycholate treatment followed by ammonium sulfate farctionation and affinity chromatography with Con A Sepharose. The absorption spectra of the preparation showed the maxima at around 410 nm for oxidized form, 422 nm for dithionite-reduced form and 422 nm for CO complex of reduced form. The cyanide difference spectrum showed a peak at 431 nm and a trough at 403 nm. This preparation was contaminated with little cytochrome b5 and it was shown that the TPO preparation was able to be used for the following spectrophotometric experiments. The addition of H2O2 to the TPO preparation induced the characteristic change in the difference spectrum with a peak at 430 nm and a trough at 407 nm, which was gradually disappeared in a few minutes, but at the high concentration of H2O2 (35 muM) the trough at 411 nm was observed after decomposition of H2O2, accompanying loss of peroxidase activity. This deepening of the trough caused the heme degradation which was dependent with the concentration of H2O2 added and to less extent at the low concentration of H2O2 (3.5 MUM). Since the difference spectrum produced by the addition of small amount of H2O2 disappeared rapidly after the addition of KI or ascorbate and resembled the spectral change due to the formation of Compound II in the reaction of other perioxidases, it was concluded that the difference spectrum with a peak at 430 nm and a trough at 407 nm observed after the addition of H2O2 was ascribable to the formation of Compound II of TPO. Although Compound I was not observed under the experimental conditions used, the results were accounted for the presence of I bound to TPO or other endogeneous reducing agents. We tentatively concluded that Compound I and Compound II are formed in the reaction of TPO with H2O2 as well as in that of horseradish peroxidase.


Determination of the concentration of plasma proteins by density measurement.

Density measurement with a commercially available instrument (DMA 46, PAAR) was studied as an alternative method to the time-consuming and sometimes expensive methods currently in use for the determination of protein concentrations. The method is very useful for protein estimation in salt--and alcohol--free albumin bulk solutions and as an additional end product test for albumin and immunoglobulin solutions; it may also be used in the determination of total protein in plasma.


Application of parallel line bioassay method to the potency test of anti-HBs immunoglobulin.

The parallel line bioassay method was applied to the potency test of anti-HBs immunoglobulin by radioimmunoassay (RIA) in an attempt to express the relative potency of three test samples in terms of ratio to the BoB reference preparation. By plotting the net c.p.m. readings without being divided by the 'cut-off' values against the dilution factors, both being converted to log10, parallelism between linear regression lines could be observed within wide ranges. Calibration of relative potency was possible wherever the regression lines were parallel, thus making any attempt of comparison at specified points alone, such as ED50, unnecessary.


Assay of anticomplementary activity in solutions of immunoglobulins.

A method of estimating the inactivation of complement by immunoglobulins, based on a hemolytic assay, is described. During a three year period of routine assays the investigation of standard immunoglobulin solutions of high and low anticomplementary activity confirmed the reliability of the method. Since the activity of complement is largely dependent on the ionic strength, the salt concentration of the test solution must be carefully adjusted. The specific complement inactivation per gram immunoglobulin CI50/g is proposed as a parameter to compare the anticomplementary activity of immunoglobulin solutions.


Purity, anticomplementary activity, and viral antibody profile of NIG preparations for intravenous and intramuscular use.

16 normal immunoglobulin preparations of 4 different manufacturers for intravenous and intramuscular use were tested with regard to purity, anticomplementary activity, and viral antibody profile. Besides IgG, which accounted for the main part of the protein content, small amounts of IgA, IgD, and IgM were found. IgE was detected in every batch studied. Some contained IgE-concentrations, which are usually seen in atopic persons. Anticomplementary activity was detected in preparations for i.m. application but surprisingly also in some preparations for intravenous use, a fact which may possibly cause allergic or anaphylactic reactions. The investigated batches showed almost the same variety of viral antibodies and only slightly differing antibody titers. Antibodies to vaccinia virus could not be demonstrated.


Toxicity of factor IX concentrates in mice.

Mice to which Factor IX concentrates are administered intraperitoneally at high doses (approximately 1000 u/kg) die between 4 and 18 hours thereafter. Histologic examination reveals hemorrhage in the lung and liver. Death is prevented by reduction of the concentrate with dithiothreitol but not by treatment of the preparation with enzyme inhibitors (diisopropyperosphorofluoridate, soybean trypsin inhibitor, heparin). Prothrombin, devoid of Factors VII, IX and X, also is lethal to mice. Activation of the Factor IX concentrate by insolubilized Russel's Viper Venom (NAPTT 1/5000 less than 20'') resulted in decreased mortality when compared to controls (4/30 vs. 11/20). These experiments indicate that some of the adverse effects of Factor IX concentrates may result from high levels of zymogens rather than the presence of small amounts of active enzymes. Intensive animal testing might be indicated for routine quality control of Factor IX concentrates.


A test method for the absence of thrombogenicity in factor IX complex.

Consumption coagulopathy was developed in three cases of liver cirrhosis and a case of haemophilia B upon administration of Factor IX preparations which had been tested by the manufacturers for the absence of active clotting factors according to the Japanese Minimum Requirements for Biological Products. By employing the TGT determination, however, active clotting factor(s) could be detected in some of the preparations used in these cases. Accordingly, it was found necessary to modify the current test method in the Minimum Requirements by introducing calcium ion into the test medium. There was no correlation between the clotting activitiy detected by our modified test method and the Factor IX potency of the product. Addition of heparin did not significantly influence the results in our modified method.


Gel chromatography applied to quantitation of components of IgG preparations.

Investigations were undertaken to standardize gel chromatography for quality control of IgG preparations. Great importance was attached to convenience of procedure. With application of the optimal type ofgel, Ultrogel AcA 34, the influence on quality of separation of various experimental parameters such as pH and ionic strength of the eluant, volume and concentration of the sample, flow rate and direction, and temperature were tested. Various methods of evaluation were applied. According to the demands, quantitation of components may be performed by collection of fractions and determination of peak volumes and protein concentrations, by determination of peak areas on absorbance recordings, or automatically by use of an integrator or a computer. The results obtained by gel chromatography are discussed and compared with the results of analytical ultracentrifugation.


Testing for potential thrombogenicity of prothrombin complex concentrates.

The methods used in the author's laboratory for testing for potential thrombogenicity in prothrombin complex concentrates are described and compared critically. Alternative methods of presenting the results of these test systems are discussed. Changes in fractionation procedure which have resulted in improved performance of concentrates in one or more of the tests are discussed.


Gel filtration in Sephadex G200 for the control of preparations of human immunoglobulin.

Gel filtration in Sephadex G200 has been used in the quality control of normal and specific immunoglobulins at the Blood Products Laboratory, Elstree, for more than eleven years. Eluting from a 140 cm column, polymerised and aggregated protein is excluded by the gel, followed in order of elution by fractions containing the IgG dimer and the monomer. After these, smaller protein impurities such as albumin are eluted and then fragments from the proteolytic breakdown of IgG. On a single chromatogram, possibly five distinct peaks may be recorded and their components quantified. Gel filtration in Sephadex G200 provides an alternative to sedimentary boundary ultracentrifugation, by which fragmentation of IgG can be more readily observed. In addition, the fraction containing the polydisperse aggregates excluded by the gel provides an index of adverse treatment of immunoglobulin during preparation and storage.