Antibodies to basal epidermal cells in patients with basal or squamous cell carcinoma. Circulating antibodies directed against cytoplasmic antigens of basal cells of normal human epidermis have been reported in association with various skin diseases. Since they have been found in patients with malignant melanoma it was proposed to investigate whether these antibodies are associated with other epidermal tumors such as basal or squamous cell carcinoma. With indirect immunofluorescence it was shown that sera from patients with basal (20%) and squamous cell carcinoma (16%) contained these antibodies. They could be demonstrated by cytoplasmic fluorescence of the basal epidermal cell layer. These antibodies occured almost as frequent as in malignant melanoma sera. However they do not seem to be tumorassociated as they were found in sera from patients with various benign dermatoses, with tumors of extraepidermal origin and even sporadically in sera from natural antibodies which were identified by cytoplasmic fluorescence of the outer epidermal cells, suggesting two antigenically different epidermal cell populations.


Melanin-producing dendritic cells and histogenesis of malignant melanoma. In a total of 70 malignant melanomas we searched for dendritic-branched fluorescent pigment cells. Hereby we found that dendritic-branched tumor cells are especially characteristic in cases of lentigo maligna. In the flat parts of these lesions, these cells are the predominant cell type. Dendrites in the pseudonests or nodular parts of lentigo maligna can only seldom be detected. The prevailing cell type in superficial spreading melanoma and in primary nodular melanoma is the round or oval unbranched tumor cell. In some cases of nodular melanoma, cells with short dendrites could be seen. In superficial spreading melanoma, dendritic tumor cells could be observed particularly in such tumor parts, in which the malignant cells were scattered between the keratinocytes. Melanocytes can evidently produce dendrites between cells of the sebaceous gland. In the marginal parts or in parts of regression of some superficial spreading melanomas, a great area of dendritic tumor cells could also be detected in the basal parts of the epidermis. Altogether, however, in superficial spreading melanoma and in nodular melanoma they occur only rarely. Dendritic-branched cells are also visible in lymph-node metastases of SSM and NM. The fact that the dendritic tumor cells can be observed in all 3 types of tumors (according to Clark and coworkers) gives a rise to a new discussion of the dualistic theory of melanoma-histogenesis of Mishima. Although this theory could not be disproved, up to now on the basis of the present results, an unitarian development of all types of mnelanoma from melanocytes seems to be possible.


Cutaneous leishmaniasis. Ultrastructural study of 3 cases. Ultrastructure study of specimens taken from 3 patients with cutaneous Leishmaniasis showed parasites within macrophages. None were found in extracellular locations. Within the macrophages cytoplasm, parasites were located either lying free in the cytoplasm or inside intracytoplasmic vacuoles. Parasites in various stages of degeneration were found both lying free in the cytoplasm or within vacuoles. The parasite-macrophage relationship and the mechanism by which the host reacts toward the parasite are discussed.


Biochemical and physiological parameters on the healthy skin surface of persons with candidal intertrigo and of persons with tinea cruris. Biochemical and physiological tests were carried out on the skin surface of 20 patients with candidal intertrigo and 27 patients with tinea cruris. In all patients the test areas were free of efflorescences. The same tests were performed in 39 and 27 resectively healthy test persons of the same age and sex. The following striking findings came to light: 1. There was a significant decrease in the percentage amount of squalene in the skin surface lipids of the moniliasis group as compared with the control group. 2. There was a significant decrease in the reducing substances in the so called water solubles obtained with the phenol sulfuric acid method in the moniliasis group. The same results were obtained when only those moniliasis patients who were definitely not suffering from diabetes mellitus were taken into account. This is presumably a question of a reduction in the bound carbohydrates. 3. There were significantly more amino acids extractable from the skin surface of the tinea cruris patients than of the control persons. These results point to important predisposing factors for the susceptibility to candidal intertrigo and tinea cruris respectively.


Effect of anti-inflammatory drugs on sulphated glycosaminoglycan synthesis in aged human articular cartilage. The anti-inflammatory drugs, sodium salicylate, indomethacin, hydrocortisone, ibuprofen, and flurbiprofen, were examined for their effects on sulphated glycosaminoglycan synthesis in aged human cartilage in vitro. Cartilage was obtained from femoral heads removed during surgery and drug effects were found to vary significantly from one head to another. Statistical analysis of the results showed that sodium salicylate exhibits concentration-dependent inhibition of glycosaminoglycan synthesis over the concentration range used. Indomethacin, hydrocortisone, and ibuprofen, at concentrations comparable to those attained in man, caused a statistically significant depression of sulphated glycosaminoglycan synthesis in cartilage from some femoral heads but not others, reflecting the variable response of human articular cartilage to anti-inflammatory drugs. Sodium salicylate and indomethacin at higher doses produced significant (Pless than 0-005) inhibition of sulphated glycosaminoglycan synthesis in all femoral heads studied. The results for flurbiprofen were less conclusive; this compound appears not to inhibit glycosaminoglycan synthesis over the concentration range used.


Defective responsiveness to ascorbic acid of neutrophil random and chemotactic migration in Felty's syndrome and systemic lupus erythematosus. Polymorphonuclear (PMN) leucocytes from 4 patients with untreated systemic lupus erythematosus (SLE) showed defective random migration (P less than 0-05) and depressed chemotactic responses to C5a and kallikrein (P less than 0-01) compared to PMN leucocytes from normal subjects, or patients with rheumatoid arthritis (4) or Felty's syndrome (4) when examined at a standardized cell concentration with a micropore filter radioassay but not with a conventional Boyden technique. Normal in vitro enhancement of PMN leucocyte random and chemotactic migration by sodium ascorbate was absent in SLE and Felty's syndrome, but sodium ascorbate gave normal stimulation of hexose monophosphate shunt activity in the PMN leucocytes precluding a defect in ascorbate transport.


Distribution of yttrium 90 ferric hydroxide colloid and gold 198 colloid after injection into knee. Thirteen knees were injected with yttrium 90(90Y) ferric hydroxide colloid, and 12 with gold 198(198Au) colloid for treatment of persistent synovitis. Retention in the knee and uptake in lymph nodes and liver were measured by a quantitative scanning technique. There was no significant difference in the retention in the knee of the two different colloids. A tendency towards higher lymph node uptake was observed with 198Au compared with 90y. The inflammatory activity of the knee at the time of treatment may have influenced the subsequent lymph node uptake of 198Au, but not that of the 90Y, nor the overall leakage of either from the knee. 90Yferric hydroxide colloid was retained in the treated knee at least as well as other colloids which have been used for this purpose.


Chondrocalcinosis after parathyroidectomy. In this retrospective study of 57 patients with primary hyperparathyroidism who underwent parathyroidectomy, the overall incidence of chondrocalcinosis was 40%. Neither joint symptoms nor chondrocalcinosis regressed after the operation. In several patients the condition appeared to deteriorate both clinically and radiologically after the operation, while in a few both the chondrocalcinosis and the associated symptoms first appeared some time after the operation.


HLA-B27 antigen in diagnosis of atypical seronegative inflammatory arthropathy. Eighteen patients with an inflammatory pauciarticular peripheral arthropathy not typical of any known entity showed an asymmetrical pattern of disease, with a predilection for the lower extremities. Destructive joint changes and deformities were absent (mean follow-up 10 years). Although spondylitis and sacroiliitis were absent on x-rays, HLA-B27 antigen was found in 8 patients (44%). This antigen and similar joint symptoms are also found in other forms of arthritis.


Sparing effect of  hemiplegia on tophaceous gout. The sparing effect of hemiplegia on the development of tophaceous gout is described. The useless upper limb had no tophaceous deposits and the partially paralysed lower limb had only limited urate deposits. Disuse was presumably the major contributor to the limited deposition of urates on the paralysed side.


[Ripening of Haden and Kent mangos covered with Tag wax in different ripening stages]. Mangos at different maturity stages were waxed with TAG and allowed to ripe at 23 degrees C and 16 degrees C, with a relative humidity of 60-70%. Unwaxed fruits held under the same conditions were used as controls. Results showed that in both varieties the waxing of the fruits extended their shelf life, improved their appearance and reduced their weight losses. Some internal characteristics like color and pH were also improved in waxed fruits, but TSS tended to be reduced in those treated fruits. It was concluded that maximum shelf life extention and best improvement in the internal qualities were achieved when fruits at the green maturity stage were treated. The significance of these findings and their possible application are discussed.


[Effects of Flavorseal and Tag waxes on the ripening and texture of Kent mangos]. Mangos KENT at the green maturity stage were waxed with Tag or Flavorseal and were allowed to ripe at 23+/-2 degrees C and 16+/-1 degrees C. The relative humidity was 60-70%. Unwaxed fruits held under the same conditions were used as controls. It was found that both waxes prolonged the shelf life of the mangos. The results also showed that Tag treated fruits developed their internal and external coloration normally, whereas mangos with Falvorseal coating did not develop their external coloration nor their red internal coloration. The content of Total Saluble Solids (T.S.S.) was higher in control fruits than in Tag treated fruits but it was lowest in Flavorseal coated mangos. As evident from the penetration and compression tests, the texture of Tag treated fruits was softer than the texture of the control and Flavorseal treated mangos. However, the texture of the control mangos was also much softer than the texture of the Flavorseal treated mangos. The patterns of the penetration and compression curves were similar in control and TAG treated fruits while in Flavorseal coated mangos the curves were uncharteristic. The significance of the results is discussed and a possible explanation for the differences between the effects of the two waxes is given.


[Bromatological study of protein concentrates from Sardinella aurita and Tilapia melanopleura. II. Analysis of minerals]. The calcium and phosphorus of fish protein concentrates (FPC) prepared from whole sardines or tilapias have high biological availability, but the mineral fraction as a whole did not support the normal growth of young rats. Fluoride absorption and utilization is low; the amount retained in the carcass of rats depends on the amount given in the diet according to the equation: Y=0,0672X + 0,202 Y=mg of fluorine retained X=ppm of fluorine in the diet In case of sub-optimal levels of mineral ingestion, fluoride fixation increased proportionally to the deficiency. Our results demonstrate that does not seem necessary to reduce the amount of fluoride in FPC intended for human consumption, at least for people without mineral deficiency in the diet. Results concerning the Ca, P and F balance in rats are presented.


[Bromatological study of protein concentrates from Sardinella aurita and Tilapia melanopleura. III. Effects of potassium suplementation on the biological value]. Bromatological study of protein concentrates from Sardinella aurita and Tilapia melanopleura. III. Effect of potassium additions on the biological utilizations. The authors studied in rats, the effect of suplementation of fish protein concentrates with Ca, P, K, or Na. Only the addition of K, but not of Ca, P, or Na resulted a better utilization of protein, showing to be the major deficiency of the mineral fraction of the concentrates.


[Evaluation of the quality of proteins of various vegetable grains used in various biological methods]. The protein value of different legume foods was studied in weanling rats by determining Net protein Utilization (NPU), Protein Efficiency Ratio (PER), Biological Value (BV), Nitrogen Balance (NB) and digestibility of the protein. NPU was determined by carcass analysis and by nitrogen determination in one of the hind legs of the experimental animals. All samples were evaluated at a 10% protein level in the ration, with and without a methionine suplement. The effect of protein concentration in the diet on the protein quality determinations used was also studied in the black bean (Phaseolus vulgaris) sample. Results showed a good correlation among the several values of protein quality evaluation determined by the different methods used, indicating that any of these procedures can differentiate protein quality between samples. Likewise, it was confirmed that NPU can be as accurately calculated by nitrogen carcass analysis as by determining the nitrogen content in the hind leg of the animals. Due to its low cost and the small amount of sample required for the determination, the modified NPU method using the traditional 10-day experimental period could be very useful in determing NPU in programs dealing with the nutritional improvement of food legumes.


[Consumption of selenium in the city of Caracas in comparison with other cities of the world]. Ingestion of selenium in Caracas, compared with some other cities. Fourty-two different food products acquired in Caracas were analized for their selenium content and the results compared with those reporte in food from Beltsville, Md. When the selenium level of a normal diet of 2200 cal is calculated, a selenium ingestion of about 220 mcg/d is found for Caracas, compared with several diets from Canada shows that the they contain from 1/3 to 2/3 of the amount of selenium as similar as similar diets in Caracas.


[Self-strangulation?].

[Self-strangulation?]. A sixty year old man was found stangled to death with a part-elastic belt. As the ecchymosis, which are typically found above the mark of strangling, were missing, considerable doubt emerged at first as to the cause of death and to a suicide by strangling. Measurements of the elasticity of the instrument of strangulation, of the size of the neck and the head, as well as self-experiments aimed at reconstruction the circumstances had, in pathophysiological respect, the result of a method of self-strangulation similar to a typical hanging. The remaining circumstances also point to a suicide.


[Urocytogram and precocious puberty].

[Urocytogram and precocious puberty]. After a review of the technique of urocytogram the authors report the results obtained in 39 children between the ages of 3 and 12 years. Before the appearance of the first signs of puberty, urocytograms are similar in both girls and boys. Differences were seen in seven cases of male or female precocious puberty. Urocytograms are also different in patients with incomplete precocious puberty but less marked than in those with true precocious puberty. The authors consider the possibility of repeat urocytograms during the supervision or treatment of incomplete and complete precocious puberty.


[Skinfold thickness measurement in children. The technic and normal values]. In a group of 3,644 healthy controls aged 0 to 19 years and 273 obese children the reference values for triceps and subscapular skin fold thckness were established. The technique of measurement of skinfold thickness is described. This is simple and may be used in routine practice as a measurement of the adipose cell mass. However, measurement of only the subscapular skin fold thickness of which the values are less scattered than triceps, was sufficient to measure the development of adipose tissue in children up to and including puberty. Those subjects whose skinfold thickness is above the upper limit of normal are considered to be obese.


[Localized hemangioma of the bronchial trunk of the left lung in a 2-month-old infant]. A 2 month-old infant was admitted because of respiratory distress and thoracic assymetry. X-ray films revealed a distension of the left lung. A pneumo-angiography showed a cardiac silhouette displaced to the right and a normal vascular tree of the left lung. The rapid impairement of the clinical picture led to an exploratory thoracotomy. A tumor at the level of the left hyparterial bronchus was found. The histologic characteristics of the tumor were those of a hemangioma. Within the spectrum of broncheal tumors, hemangioma belong to benign mesenchymatous tumors and have a good prognosis providing that their removal is complete.


[Pharmacokinetic data on phenobarbital. Study on the influence of gestational age and dysmaturity]. Blood levels of barbiturate after a single intramuscular injection of 10 mg of phenobarbiitone have been plotted against time in 15 neonates at term of whom five were normal and 10 were dysmature, and 15 pre-term infants between 32 and 34 wesks gestation, of whom 12 had a satisfactory weight and 3 were dysmature. The analysis of the experiments according to a mathematical model gave the following results. Half life of absorption +/- 1 S.D. 9.55 HR +/- 6.97 for the term neonates and 11.12 hr +/- 7.61 for the pre-term infants. The difference is not significant. Half life of elimination +/- 1 S.D. 118.06 hours +/- 64.79 for the neonates of term and 110.49 hr +/- 37.19 for the pre-term infants. The half life of elimination is much shorter in the pre-term dysmature infants 74.10 hrs +/- 40.12. The individual variations are important but the differences between the groups of neonates studied are not great. This permits re-grouping and calculation of a half life of elimination for the whole group of 114.28 hr +/- 28.05. The risk of over dosage and the large individual variation in the metabolism of phenobarbitone in the new born lead us to recommend checking the blood levels when giving repeated doses.


[Determination of the amino acid spectrum as an aid in the up-dating of skeletal findings]. Estimation of the age of skeletal remains is usually based on subjective evaluation of the morphological grade of decomposition. This assumes, that the investigator is provided with pertinent experience. Experimentally elaborated physical and chemical methods proposed for objectification of the statements did not yet assest until today due to lacking temporal correlation. Our investigations were stimulated by observations on fossil bones indicating that the collagenbound amino acids diminish very slowly dependent on geological age. Samples of 53 skeletons of five to 2500 years were pulverized and subjected to acid hydrolysis. The amino acids released from bone collagen were separated by means of thin-layer chromatography and detected by isatin and ninhydrin reagents. The dye complexes of alanine, hydroxyproline, proline, tyrosine, and valine were measured densitometrically and quantified through calibration curves. We established, that the concentrations of the amino acids exhibited a time dependent reduction as equivalent of a loss collagen. The temporal correlation is very narrow (r less than -- 0,8) and sufficient for datings within plus/minus ten years of the more recent bones. Environmental conditions during post-mortem decay seem to have little influence on this process.


[A new case of congenital short small intestine with intestinal malrotation]. The 11th case of congenital short small intestine malrotation is described. Its clinical features associated diarrheic episodes and subobstructive symptoms. Recovery suggests that the functional adaptation of congenital and surgically-induced short small intestine may have the same quality. Neither anatomical (presence or absence of pyloric hypertrophy) nor clinical variants (obstructive and diarrheic forms) challenge the nosologic unicity of this syndrome, whose genetic character is unquestionable in most cases.


[The orthographic quotient. Quantitative study of the free text for dyslectics]. The orthographic quotient (O.Q.) is the rate per cent of correctly spelled words in spontaneous writing of dyslexic children. O.Q. was studied in 31 children. It was found a constant O.Q. in each child and a positive correlation with the reading level (p less than 0.001). O.Q. may be regarded as a new quantitative characteristic of dyslexiadysorthography. Contrary to other tests, O.Q. does not require any standardized material and may be repeated ad libitum. It might be applied to: 1. the diagnosis of dyslexia and the assessment of progress during remedial teching; 2. the early detection of dyslexia in primary schools.


[Introduction to perinatal pharmacology]. Attention has been focused on the specific pharmacokinetics and clinical pharmacology present during the perinatal period following clinical accidents due to high concentrations of oxygen, vitamin K2, sulfonamides, novobiocin, chloramphenicol or drugs of the tetracycline family. The mechanisms of these therapeutic accidents are subject to certain parameters: renal excretion, inactivation systems, binding to plasma albumin, affinity of the tissues for the drugs, intra and extra-vascular compartment distribution, and pharmacogenetics. The special cases of maternal passage of drugs, either during labor or after birth in the milk given to the newborn, are discussed. Lastly, the ethical problem of "therapeutic trials" in newborns are analyzed.


[Bilirubin metabolism in the newborn. Recent progress]. The main stages of bilirubin metabolism and the present state of neonatal hyperbilirubinemia are reviewed. The heterogeneity of substrates for bilirubin synthesis and the regulation of the heme-oxygenase activity are stressed. The importance of albumin for the bilirubin transport and the factors modifying this transport are studied. The detection of free bilirubin and reserve bilirubin binding capacity in neonatal jaundice is discussed as a method to estimate the risk of kernicterus. The induction of bilirubin glucuronyl transferase activity with some drugs and the phototherapy are effective in the treatment of hyperbilirubinemia; their values are discussed from the most recent reports.


[Experimental results with different fractionation regimes and low-LET irradiation of mice (author's transl)]. The effect of several fractionation regimes was examined by the model of mammary tumour in C3H-mice. The best results were observed with several fractionations daily. No curative effect was obtained by means of a total dose of 6000 rd achieved with all fractionation regimes. The results of investigations underline the importance of considering cellular proliferation kinetic processes in malignant tumours in order to optimize the temporal distribution of radiation dose.


[Syngeneic ascitic mouse sarcomas-test systemt in chemotherapy (author's transl)]. The establishment of two syngeneic mouse sarcoma lines is described, which were induced with benzo[a]pyrene originally as solid tumours growing in the subcutaneous region, and which subsequently changed to the ascitic form. Successive i.p. transplantations produced progressive increases of malignancy. The same progressive tendencies were found for metastatic trials carried out in parallel with the i.p. transplantations. The use of both ascitic tumour cell lines for standardized chemotherapeutical screening systems is discussed.


[Clinical examination of the conception '86 of "Canoer Multistep-Therapie" (CMT) by M. v. Ardene. First communication: the standard, the control-parameters, and the course of therapy (author's transl)]. It is reported about the clinical testing of the "Cancer Multistep Therapy" (CMT) by M. von Ardenne (CMT Concept '74). It was made in 1974/75 on 42 tumour-patients. At first we demonstrate in this publication clinical and paraclinical findings, which may be relevant for the therapy-standard, but the results of the therapy-efficiency will be given later. On the base of findings and observations we verify a program-true employment of the CMT Concept '74 and we demonstrate the pathophysiological reaction-patterns in connection with the longterm-hyperglycemia (glucose-utilization, spontaneous hyperthermy with tachycardia, spontaneous leukocytosis with typical deviations in the hematogram). These reaction-patterns are described particularly as "general adaptation syndrome" (Selye). Further we give an assessment of compatibility, tolerableness, and practicability of the CMT.


Sexual attitudes and behavior patterns during and following pregnancy. Changes in sexual attitudes and behavior patterns that might be associated with pregnancy and childbirth were investigated. The subjects were 216 women seen by a group of obstetricians and gynecologist in office practice. While there was a wide range of individual responses, in general a decline in sexual interest, activity, and satisfaction was reported as pregnancy progressed. The desire for body contact remained at a very high level throughout. Some changes in body zones regarded as most erogenous during the course of pregnancy were indicated. Following childbirth the women's sexuality seems to have returned to a normal level. The degree of liberal vs. conservative attitudes toward sex expressed by these women was not affected by the pregnancy, nor did this dimension relate significantly to other behavioral variables.


[Morphological studies of gastric arteries in patients with gastro-duodenal ulcer]. Microscopic lesions of the gastric arteries have been studied in 21 patients subjected to resection for gastroduodenal ulcer. The left gastric artery, particularly in patients with gastric ulcer, presents lesions to the intima and media and these can lead to obliteration of the lumen, classifiable as atherosclerotic disease. The seriousness of the lesions would seem to be independent of sex and age. In some cases gastric artery lesions were compared with those to the arterioles in the vicinity of the ulcer. At least as far as the gastric district is concerned, there would not appear to be any gravity correlation between large and small vessels. Some aspects of the vascular pathogenesis of gastroduodenal ulcer are discussed.


Love, sex, permissiveness, and abortion: a test of alternative models. Alternative models of premarital abortion permissiveness (PAP) are tested with a two-state sample of single college women. The first model proposes that number of times in love (Love) leads to coitus (Sex) and that coitus increases premarital sexual permissiveness (PSP), which in turn leads to greater acceptance of abortion (PAP). The second posits a causal path from Love to PSP to Sex to PSP. The data appear to be consistent with a revised third model. The paths between Love and PAP and between Love and PSP are negligible, indicating that the effects of Love on PSP and PAP are mediated by Sex. The revised model suggests that as number of times in love increases so does coital experience and that coitus has both a direct effect on PAP and an indirect effect through PSP.


A typology of transsexualism: gender identity theory and data. An analysis of postoperative data obtained from 42 male-to-female transsexuals showed them to fall into three distinct categories: homosexual transsexuals, asexual transsexuals, and heterosexual transsexuals. Additonal data covering the areas of gender vs. sexual dysphoria, surgical details, background characteristics, sexuality-related behaviors, and postoperative adaptation further differentiated these groups and suggest that the categorization is theoretically meaningful and relevant to understanding the causes and course of the transsexual phenomenon. An extensive developmental theory is suggested to account for both differences and commonalities among the different subgroups of transsexuals, as well as among these and related groups, such as effeminate homosexuality and transvestism. The theory is sufficiently specific for making empirical predictions.


[Etiopathogenesis of juvenile osteochondrosis of the hip. Apropos of 2 cases observed in bicorial twins]. Two cases of juvenile osteochondrosis of the hip in biovular twins form the basis for bibliographical research into the aetiopathogenesis of the disease. Although pathogenesis is now known to be a more or less severe trophic disturbance of the head of the femur caused by faulty arterial flow through the local circulation, the prime causes of such circulatory arrest remain doubtful. Critical examination of studies carried out on the question would seem to point to a Perthes disease aetiology which would fit most of the more authoritative theories.


[The effect of the raw protein supply on the lysine requirements of young pigs of 12-40 kg]. 3 N balance trials were carried out with pigs weighing 12-40 kgs to test values for the lysine requirements of pigs that had been obtained in feeding trials with rations containing varying levels of crude protein. The present trials used rations containing 17%, 15% and 13% crude protein; the corresponding lysine percentages were 0.89, 0,79 and 0.62. The N balances revealed significant differences between the three crude protein levels amounting to 1.50 g, 1.36 g and 1.05 g per kg liveweight 0,67.


[Lysosomal enzymes in the neoplastic pathology of the uterine cervix. Study of acid phosphatase in the diagnostic test of vaginal secretions in patients with pre-invasive and invasive cancer of the uterine cervix. I. Methods and preliminary experiences]. Gomori's method was used in the detection of positive -- or negative -- phosphatase granulations and their intracellular locations in vaginal secretions from 10 normal women and 20 patients with cervical neoplasia in various stages. Positive-phosphatase granulations were absent in normal subjecta and those with advanced neoplasia, whereas mainly large and medium-sized paranuclear positive granulations were a prominent feature of epithelial cells from patients from state 0-ToNoMo carcinomas. Further details will be given in a second note.


[Pulmonary arteriovenous fistula within the picture of congenital angiodysplasias. Clinical case]. The aetiopathogenesis, clinical picture, diagnosis and treatment of pulmonary arteriovenous fistula are examined in the light of a case observed at the Florence University Surgery Clinic in 1970 and marked by multiple truncular fistulae at the middle and inferior lobes of the right lung. The condition is seen as an instance of congenital angiodysplasia with arteriovenous shunt. A clear distinction is drawn between true fistulae, as seen in the reported case, and angioma and aneurysm. Stress is laid on the importance of radical management, though it is conceded that surgery should occasionally be renounced in the absence of symptoms, or where lung hypertension is particularly marked.


[Lysosomal lesions during a course of acute experimental muscular ischemia. Protective effect of a polypetitide proteinase inhibitor]. Three groups of rabbits were used: a) with acute ischaemia in a rear limb; b) with acute ischaemia in a rear limb and treated with i.v. 100,000 KIU of a proteinase-inhibitor polypeptide extracted from ox lung; c) normal controls. Acute ischaemia was obtained by ligature of the ipsilateral common iliac, external iliac, inferior epigastric and femoral arteries. Soluble and total activity of 3 lysosomal enzymes (cathepsin, acid phosphatase and N-acetyl-glucosaminidase) were determined in gastrocnemius muscle from all 3 groups. The mean ratio between bound and soluble activity for all 3 enzymes in normal gastrocnemius muscle was higher than in ischaemic muscle, but not significantly different from that in ischemic muscle of animals treated with the polypeptide. Furthermore, this ratio in ischemic muscle was significantly lower than that of ischemic muscle of the rabbits treated with the polypeptide. These data suggest that the polypeptide offers protection against lysosomal lesion in the course of experimental ischaemia of the skeletal muscle.


[Secretion of radioactively labeled amino acids into the digestive tract. I. Report 14C labeling of the pancreas after subcutaneous injection of 14C-leucine in rats]. 4 different groups of experimental animals (rats) were injected 14C leucine at 4 different levels of specific 14C activity. Groups A and B received a dosis of leucine corresponding to the daily leucine requirements of the animals. The specific 14C activity was high in group A and low in group B. Groups C and D were injected a leucine dosis at a level of 10% from the groups A and B. the specific 14C activity being again high (group C) or low (group D). The results showed that a true evalution of the experiment could lnly be achieved if the values were related to the true rate of labeling (DPM/mg N or specific 14C leucine activity). The "percentage of the injected dosis" proved to be unsuitable for the analysis of the experimental results when taken as reference frame. The levels of specific 14leucine activity in free plasma leucine were in the same order as the specific activities of the injected solutions (C greater than A greater than D greater than B) whereas the order of specific 14C leucine activities in the TCE-soluble fraction of the pancreatic gland (TCE-trichloroacetic acid) was found to be A greater than C greater than B greater than D. This was due to the higher rate of 14C-leucine transfer into the protein fraction in the animals of group C and D receiving a normal supply of leucine. In these groups an equilibrium in specific 14C-leucine activity between the TCE-soluble and TCE-precipitable fractions was achieved 20-30 mins after injection. A relatively high degree of labelling of the pancreatic protein was noted as early as about 2 minutes after the injection of 14C-leucine. This indicates that in studies on amino acid absorption the secretion of those amino acids that are first absorbed and then resecreted via the pancreatic gland should always be taken into account.


[The effect of the raw protein supply on the lysine requirements of young pigs of 12-40 kg. 1. Report. Feeding studies with wheat-peanut extraction residue rations]. 10 feedings trials were carried out with a total of more than 500 pigs weighing 12 to 40 kgs. To investigate the lysine needs of growing pigs, the animals were fed rations of wheat + extracted ground nut meal. Different food mixtures were tested containing 5 levels of crude protein (19%, 17%, 15%, 13% and 11% of the dry feed. Within each crude protein level 6 graded lysine supplements were added to the ration. The trial showed that the lysine requirements of the weaned pigs were in a decisive measure determined by the percentage proportion of crude protein contained in the ration. The crude protein portion may be calculated (for the liveweight range tested) by using the following regression equation: y=0.28+0.075x (y=lysine requirements expressed as % of the air-dried ration; x=percentage proportion of crude protein in the ration). Rations containing only protein sources of vegetable origin, with a minimum protein content of 15%, produced the same daily weight gains (520 g) as a standard diet, if the lysine demands were met through the supplementation of synthetic lysine.


[The porosity of bone cement (author's transl)]. This paper describes a method of measuring the porosity, shape, distribution and formation of cavities in acrylic cement. We distinguish between big spherical cavities greater than 60 mu and small cavities less than 60 mu which form a variable "basic porosity" with a large number, but a minor volume of bubbles. The bubbles are formed in two different ways. The first is through introduction of air bubbles in the mixing phase by stirring the cement before the working time. These bubbles expand little by temperature rise. The second method of formation is through neoformation of bubbles by evaporation of the monomer at about 100 degrees C. This neoformation of porosity expands the cement. The rise of volume through the neoformation and expansion of bubbles is able to compensate the shrinking of cement due to the polymerization. This effect, however, cannot be currently controlled. Cement strength is influenced by the degree of porosity.


[Biological effect of bariumsulfate as contrast material in bone cement. An animal study on rabbit femora (author's transl)]. The investigations presented in this paper concern the question of toxicity of Bariumsulfate as an ingredient of bone-cement. In 6 rabbits the medullary cavity of the right femur was filled with bone-cement which contained Bariumsulfate as X-ray contrast ingredient and the left one with BaSO4-free bone-cement. 4 1/2 months post operationem the femura were investigated by histological and microradiographic methods. The status of both sides was compared. On the BaSO4-containing side regularly necrotic lamellar bone structures in the zones of osteogenesis and single disseminated necroses are found which are not caused by the primary lesion occuring during the implantation procedure. Additionally the zone of primary lesions which is found on both sides was more extensive at the BaSO4-containing side.


[Deformity and arthrosis in slipped upper femoral epiphysis (author's transl)]. X-rays of 104 slipped upper femoral epiphysis (without reduction of epiphysis and complications) showed pathological datas of the hip joints (ACM-angle, CE-angle, Hüftwert) in one third of the cases in the beginning and 60% until 12 years later. With higher degree of slip the arthrosis increase, but not the pathological datas of the hip joint.  There is no dependence between deformity of the hip and arthrosis. Apparently the articular cartilage necrosis partially determine the development of arthrosis.


[Evaluation of loosening of different total hip models and neck lengths (author's transl)]. The percentage of aseptic loosening of different types of total hip prostheses in 771 cases has been evaluated. The time of follow-up ranged between 1/2 and 6 years. In prostheses of the Charnley-Müller type of loosening was 7.8% in the Charnley-Müller-Weller type with a 2-year shorter follow-up it was 2.4%. No loosening was found during the last 2 1/2 years when the prosthesis of Beck was used, which has an increased femoral shaft-neck angle and a longer stem. When a longer neck was used, the loosening averaged 5.6%. In cases of normal neck-length it was 3.6%. From the biomechanical standpoint it seems preferable to use prostheses with an increased shaft-neck angle. Comparing the rates of loosening as reported in the literature it appears that prostheses where the femur is displaced medially (by a small femoral head or increased shaft-neck angle) show a very low rate of loosening. Models with metal sockets (McKee-Farrar and Weber-Huggler) seem to have a higher rate of loosening than models with polyethylene sockets.


[On the pressure distribution between femoral head and cup of a total hip joint prosthesis (author's transl)]. The stresses that occur between femoral head and cup in a total human hip joint prosthesis are especially calculated for the running-in phase when transition takes place between the Hertzian stress distribution and the stress distribution caused by wear. It turns out that the Hertzian stress distribution has already altered considerably after nearly 1 year of wear.


Stress and distress in middle age - The mental health of doctors. In considering subject material of "Stress and distress" in a symposium devoted to emotional probelms of middle age, who better to consider than the doctor? What effects does the stress of medical practice have on the mental health of the practitioner and his family?


The GP - doctor or middle-man? A study of his paper work. The convention of using medical certificates to assess benefit claims has grown over the years to a complex situation. The certificate serves as written testimony of the patient's state of health (be it illness, fitness or a need which is based on those grounds). While the practitioner is presumed to be serving only his patient, this study suggests that he is required to act as arbitrator in many situations that are unnecessarily defined as medical.


An analysis of the activities of general practitioners. This study has demonstrated that activity sampling is a practical and sensitive method of analysing the general practitioner's task. The data so gathered can be used in planning and devising innovations in practice management, in planning and evaluating the use of allied health professionals for non- diagnostic tasks, for assisting in the definition of objectives for medical education more clearly, and finally, when the appropriate indices have been developed, for evaluating the quality of care provided.


A survey of human sexuality teaching in the medical course. In recent years, the curriculae of some Australian medical schools have been extensively revised. However, being revisions, as distinct from reconstructions, new subjects were only added if they were thought of at the time; for example, electives. Still more recently, there have been cries for undergraduate medical education to involve itself more with the common problems typically facing the family physician. This has paralleled the wave of verbal opinion by medical students, doctors, and allied health professionals, that the field of "Human sexuality" should be included in the medical curriculum as an integrated but distinct entity. The survey presented in this article was carried out to document this opinion on paper, and to obtain information which may be relevant in proposals for establishing such a formal teaching programme.


Comprehensive management of the hyperkinetic child. Family practitioners have been and will continue to be, in all countries, "front line psychiatrists" for their communities. This fact is particularly true for child care, since child psychiatrists are so few in number, and they are hardly able to see, still less treat even the tip of the iceburg of troubled children. Parents and schools turn first of all to their family physician. Today, quite commonly, the defined problem is "he's hyperactive" -- a term which has appeared in educational, medical, scientific and general literature since the 1950's. A practical and comprehensive approach towards understanding and management of these children is outlined here, with the suggestion that a copy of this paper could productively be shared with parents, teachers and even the affected child, depending upon his or her cognitive level.


Principles of management of juvenile-onset diabetes. This paper discusses "juvenile-onset diabetes", which is a condition that can begin at any time during the life span. The condition is understood to mean that type of diabetes in which there is absolute insulin deficiency, and where insulin therapy is required.


Characterization of cells of the normal aortic endothelium of adult rats and changes due to endotoxin shock. I. Communication: light microscopy, autoradiography, DNA cytophotometry, and enzyme histochemistry. Aortic endothelium of normal and endotoxin treated rats was investigated in "Häutchen" preparations by light microscopy, autoradiography, DNA cytophotometry, and enzyme histochemistry. In the endothelium of control rats the following cell populations were found: 1. Endothelial cells (90% of all cells in endothelium), characterized by the alignment of the longitudinal axis of their nuclei, their light microscopic morphology, their DNA content resp. DNA synthesis, and by their enzymatic activity; 2. mononuclear cells (10% of all cells in endothelium), identified as mature blood monocytes, mononuclear phagocytes (macrophages), and as lymphocytes; 3. granulocytes (less than 1% of all cells in endothelium). The presence of mononuclear cells and granulocytes in the normal endothelium appears to be the expression of a physiological repair process in endothelium. After endotoxin application one can observe an endothelial damage and then a repair process. During the repair of endothelium the following processes occurred: An increase of DNA synthesis particularly of endothelial cells, an aggregation of platelets on the endothelial surface, an increase in number of granulocytes and at last an increase in number of activated mononuclear cells. Participation of mononuclear cells in the physiological as well as in the pathological repair process of endothelium was shown by the present investigation. The process of transformation of mononuclear cells into endothelial cells is discussed at the same time.


[The presence of epithelioid glomus structures in the lymph nodes of excised mammary carcinomas (author's transl)]. A report is presented on the demonstration of epithelioid cell clusters within the axillary lymph node group, which - in the absence of knowledge of their structural peculiarities--could be taken for micrometastases of a solid mammary carcinoma. The lymph node material from 400 extensively excised mammary carcinomas with obligatory lymphonodectomy was available for this study. Microscopic inspection of successive sections at intervals of 100 mu was performed following fixation and optical demonstration of all lymph nodes in a picric acid medium. This procedure led to the discovery of epithelioid cell clusters in 18 out of the 400 cases examined. As regards their appearance, with direct allocation to the local vascular stroma, these clusters must be regarded as glomus-like structures. In no case could metastases of a mammary carcinoma and the epithelioid cell foci mentioned here be demonstrated either next to each other or combined. In appearance they are remarkably similar to the already known glomus structures of other sites. The presence of such structure in lymph nodes is most likely attributable to micromorphous malformation of vessels in the sense of hamartia. From a clinical point of view, their fundamental significance consists in the necessary differentialdiagnostic demarcation against metastase of solid carcinoma in the region of origin of the lymph.


Quantitative histological aspects of liver cirrhosis. In human liver cirrhosis and other liver diseases relatively large nuclei are found with finely meshed chromatin and large nucleoli which do not occur in liver of individuals without liver disease. The number of tetraploid nuclei near central veins and portal areas in normal livers is not statistically significantly increased in comparison with the surrounding areas. The percentage of diploid nuclei in these livers varies from 80-95 per cent. There are definite differences in the number of adjacent microscopical fields of view with an increased degree of polyploidy when livers of normal individuals are compared with livers from patients with suspected liver disease and patients with liver cirrhosis. The increased degree of polyploidy in the last two groups can be considered as sign of regeneration.


Comparison of the effects of ethidium bromide and of ethidium bromide-deoxyribonucleic acid complex in fibroblasts cultivated in vitro. Chick embryo fibroblasts cultivated in vitro were treated with ethidium bromide (E.B.) or with DNA-E.B. complex (DNA-E.B.). E.B. (5 mug/ml) provokes morphological alterations and cell death, inhibits DNA synthesis and mitotic activity. DNA-E.B. (E.B. 5 mug/ml) is less toxic to the fibroblasts as far as cell structure, DNA synthesis and mitotic activity are concerned. DNA alone has no apparent effect on the fibroblasts. As shown by fluorescence microscopy, the lower toxicity of DNA-E.B. seems to be related to its mode of penetration into the cells.


[Serological and clinical aspects of mass transfusion]. Massive transfusion of cold blood causes hemodinamic disturbances because of its acidity, surplus of potassium and defiency of calcium. Negative influence also results from stronger viscosity of blood, from the change of the active substances of blood coagulation, change of the patient's coagulation system responding to the transfused blood, subclinical troubles in this system according to the degree of the shock, reduced ability of oxygen transport caused by viscous blood, poor microcirculation etcr. Arrythmia, bradycardia, fibrilation of heart ventricles, may occur. Vasoconstrictions, microembolism and other incidents may appear. The quality of preserved blood changes during the conservation is changed, potassium and ATP are the indicators for its quality. At massive transfusions blood has to be warmed, filtered through special filters, to the patient has to be given just the quality of blood he needs etcr. The transfusiologist can help the therapist with useful advices and informations for the patient's sake.


[Preparation of granulocyte concentrates from healthy donors using a continuous-flow centrifuge (CFC)]. By means of continous flow centrifuge the authors have obtained suspensions of granulocytes from the blood of healthy donors. Results of granulocytophereses are presented here with the main points in the therapy of granulocytopenic patients with granulocytes rich suspension. The mean volume of the obtained suspension was 418ml. and it contained 9,78 X 10(9) leukocytes. 70 percent of them were granulocytes, the said volume contained 48 ml of red blood cells. All the donors had been premedicated with i.v. dexamethason (4mg./kg body weight). No one of the 51 donors developed anaemia or hypoproteinemia. After the application of the protamin sulphate, however, two donors had face flushing breathlessness and gastric ache. These symptoms disappeared after the administration of corticosteroids. Although they are connected with the administration of protamin sulphate we can not explain them.


[Concentrated antihemophilic globulin in the treatment of hemorrhagic syndromes]. The author presented the general informations on the preparation of the Factor VIII and Factor IX concentrates, on their use and side effects. Special attention was paid to the resolutions of the Scientific group on the Inherited Blood Clotting Disorders, in the meeting of the World Health Organization, which was held in Geneva, 1972.


An electrophoretic polymorphism in the serum transferrins of Praomys (Mastomys) natalensis. Three serum transferrin phenotypes of Praomys (Mastomys) natalensis, and African rodent having a high incidence of many varieties of spontaneous neoplasms, were detected by electrophoresis and autoradiography. On electrophoresis, transferrin of Trf-K was found to be a protein that migrates slowly toward the cathode, whereas that of Trf-M migrates rapidly. Trf-KM is a heterozygous phenotype of the above two. Progeny testing demonstrated that these three serum transferrin phenotypes in Mastomys are probably controlled by two codominant allelic genes, Trfk and Trfm, at a single autosomal locus. The genotypes of these phenotypes may be Trfk/Trfk, Trfm/Trfm, and Trfk/Trfm.


Genetic study of pancreatic proteinase in mice (Mus musculus): genetic variants of trypsin and chymotrypsin. Three to five isozymes of pancreatic proteinase exist in mice, and they have been designated as bands I, II, III, IV, and V. Identification experiments of these isozymes were carried out in this study; bands I, IV, and V are trypsin, and bands II and III are chymotrypsin. Therefore, it is concluded that Prt-1,  controlling band V, is a locus for trypsin and Prt-2, controllong bands II and III, is a locus for chymotrypsin. In addition, a new locus, Prt-3, has been found. At this locus the two allelic genes, Prt-3a and Prt-3b, control the low and high tryptic activities of band IV, respectively. Prt-3 is present only in the strain Mol-A. Linkage experimentation has shown that Prt-1 is closely linked to Prt-3.


Synchronous allelic expression at the glucosephosphate isomerase A and B loci in interspecific sunfish hybrids. Allelic isozymes of glucosephosphate isomerase at the Gpi-A and -B loci were separated by starch gel electrophoresis in the warmouth (Lepomis gulosus) and green sunfish (L. cyanellus). The specific tissue distributions and developmental expressions of the GPI-A2, -AB, and -B2 isozymes were not different between these two species. The synchrony of allelic expression in normal intraspecific sunfish crosses was demonstrated by means of an electrophoretic variant at the Gpi-B locus. In embryos formed from warmouth x green sunfish hybrid crosses, the paternal GPI-A2 isozymes were first expressed at the same time in both reciprocal hybrids, at 21-25 hr after fertilization. The maternal and paternal GPI-B subunits were synchronously expressed in reciprocal hybrids just for prior to hatching. The parental allelic isozymes at both loci shoed codominant expression in all tissues of the mature F1 hybrids. These results are consistent with the absence of allelic asynchrony and inhibition in interspecific hybrids formed from more evolutionarily related species.


Mitochondrial proline dehydrogenase deficiency in hyperprolinemic PRO/Re mice: genetic and enzymatic analyses. Genetic analyses, involving backcross and F2 matings, demonstrate that the type I hyperprolinemia of PRO/Re mice is caused by an abnormal allelet at a single locus designated pro-1. Mice homozygous for this allele (pro-1b/pro-1b) posses a deficiency in the activity of component 1 of mitochondrial proline dehydrogenase. In liver mitochondria of normal C57BL/6J mice, two proline dehydrogenase activity components are demonstrable by electrophoretic resolution of Triton X-100 solubilized extracts. In mitochondria of PRO/Re mice, the activity of component 1 is not readily detectable. Residual proline dehydrogenase activity in PRO/Re mitochondria appears, therefore, to be due in large measure to activity component 2 which is more stable to incubation at 40 C, exhibits slower electrophoretic mobility, and is less reactive to menadione. Kinetic analyses demonstrate a Km (proline) for the Triton X-100 solubilized enzyme activities of PRO/Re and C57BL/6J liver mitochondria of 0.4 M and 2.9 X 10(-3) M, respectively. C57BL/6J enzyme activity is inhibited by high substrate concentration. The actins of PRO/Re liver obtained by differential centrifugation. Abnormal control of respiratory chain function in PRO/Re mitochondria appears to involve primarily proline oxidation, as indicated by the level of activity of several inner membrane enzymes.


Relationship among H-2 type, serum testosterone level, and kidney beta-glucuronidase activity in the mouse. C57BL/K1, DBA/2/K1, and backcross male mice have been analyzed for H-2 type, serum testosterone level, and kidney beta-glucuronidase activity. No associations or correlations were found among these three parameters in the backcross material.


Purification and parital characterization of two genetic variants of placental alkaline phosphatase. The two most common variants of placental alkaline phosphatase, the F and S variants, were purified to homogeneity and characterized. Their molecular weights were determined by equilibrium ultracentrifugation and sodium dodecylsufate polyacrylamide gel electrophoresis, which gave almost identical values for the two variants, 118,000 (F) and 119,000 (S). The amino acid compositions of F and S variants presented here are found to be very similar. Differences between the two variants were found in specific activity (160 U/mg for F and 250 U/mg for S), isoelectric point (IP - 4.5 for F and 4.7 for S),  sedimentation coefficient (6.5 X  10(-13) sec for F and 6.4 X 10(-13) sec for S). Thus the structural differences observed for these enzyme variants seem to affect both the active site and the protein conformation.


Genetic divergence of insular populations of deer mice. Electrophoretic variants at 28 genetic loci were analyzed in subspecies of Peromyscus maniculatus endemic to the Channel Islands of the California coast. The genetic variability within insular populations was calculated. These deer mice have relatively high levels of polymorphism for insular populations. The mean heterozygosity per individual varies in the populations, being much higher on one of the islands than the others. Nei's measure of genetic distance between groups compared on the basis of electrophoretic variants was used. His estimate of time of divergence of these groups, based on genetic distance, is applicable particularly to closely related groups. The length of time each island population has been isolated from the others was calculated and found to be consistent with the periods of isolation estimated on the basis of geological data.


Genetic effects on male mouse kidney glucuronidase activity. Kidney beta-glucuronidase activity in C57BL/K1 and DBA/2/K1 male mice differes about tenfold, C57 giving low and DBA high values. Another C57 subline, C57BL/6J, has slightly higher activity than C57BL/K1. There is an association between the kidney glucuronidase activity and coat color determined by the buff locus, which indicates that part of the variation is due to differences at the Gur locus. The bf allele per se raises the activity of the enzyme. The backcross distributions give evidence that at least one more locus is involved.


Simplifications of the derivations and forms of steady-state equations for non-equilibrium random substrate-modifier and allosteric enzyme mechanisms. The steady-state equations for "random" enzymic mechanisms (ones with alternative routes for substrate and enzyme to form enzyme-substrate complexes) are non-Michaelian and very complicated when a quasi-equilibrium approximation cannot be used. General methods for simplifying their forms and derivations are given and applied to several single-substrate mechanisms of general or topical interest. The special simplifications resulting from partial ordering of reaction mechanism, from gross inequalities of rate constants, and from special relationships between catalytic and dissociation rate constants, are considered with reference to allosteric mechanisms. Some equations mentioned, but not given here, and more detailed working out of some of those given, have been deposited as Supplementary Publication SUP 50069 (18 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms given in Biochem. J. (1976) 153, 5.


Stable thiobarbituric acid chromophore with dimethyl sulphoxide. Application to sialic acid assay in analytical de-O-acetylation. With dimethyl sulphoxide instead of butanol in the thiobarbituric acid assay for sialic acid, a non-fading chromophore with lambdamax. = 549 nm was produced in a homogeneous solution, allowing dilution of the test mixture in case of high colour yield. This test adapted well to studies on alkaline de-O-acetylation. Bovine and rat submaxillary mucins, and rabbit Tamm-Horsfall urinary sialoproteins contain O-acetyl isomers of neuramine acid that are resistant to the thiobarbituric acid assay. Alkaline de-O-acetylation converted resistant O-acetylneuraminic acid into thiobarbituric acid-reactive sialic acid, and such conversion paralleled de-O-acetylation as measured by the ferric hydroxamate method. The colour increment was similar when the alkaline treatment of bovine submaxillary mucin either preceded or followed the acid hydrolysis. Only alkaline preptreatment was effective with rat submaxillary mucin. By selecting optimal conditions for alkaline de-O-acetylation, O-acetyl isomers can be accurately assessed by the thiobarbituric acid assay.


The preparation of protected fragments of lysozyme for semisynthesis. This paper reports the development of methods for preparing tryptic fragments of hen's-egg lysozyme in an appropriate state of protection for use in the chemical synthesis of modified polypeptides. 1. We describe the cleavage of the disulphide bridges of the enzyme and the simulatneous protection of the liberated thiol groups by S-sulphonation. Lysozyme resisted the usual conditions for this reaction. We have confirmed the stability of the S-sulphonyl group to the conditions met in peptide synthesis. 2. We describe the reversible protection of the amino groups of the enzyme by reaction with various anhydrides of 1,2-dicarboxylic acids. We conclude that 2-methylmaleic anhydride and exo-cis-3,6-endoxo-delta4-tetrahydrophthalic anhydride are unsuitable for our purpose but that maleic anhydride can, in spite of certain drawbacks, be used. 3. We describe the tryptic cleavage of the thiol- and amino-protected protein and the separation of the fragments. 4. We describe the reversible protection of the carboxylic acid groups (including the specific deprotection of the alpha-carboxyl group), the imidazolyl group and the aloph-amino groups of the fragments. Several alternative groups have been evaluated for most of these purposes. The side-chain amides did not present any serious problem of libility, 5. We describe experiments on the stability of the side chain of tryptophan, both protected by formylation and unprotected, to the acid conditions needed for the deprotection of the other functional groups in the peptide. We conclude that protection of tryptophan is unnecessary. We suggest that most of the methods described are of general application in peptide semisynthesis by fragment condensation. An Appendix is included to which points 6-ll appertain...


The semisynthesis of portions of hen's-egg lysozyme by fragment condensation. 1. We have evaluated several methods for coupling protected tryptic peptides and have selected one that is satisfactory in terms of efficiency and degree of racemization. 2. We report the use of the chosen method (which must be slightly modified for each application in the light of the result of micro-scale pilot experiments) for the preparation of five fragments of hen's-egg lysozyme ranging in length from 31 to 51 residues. 3. We report methods for the complete deprotection of these fragments. They have been characterized by end-group determination, amino acid analysis and tryptic digestion.


Determination of the number and relative position of tryptophan residues in various albumins. A technique is described by which both the numbers of tryptophan residues and their approximate locations in the peptide chain of a protein can be determined by cleavage with N-bromosuccinimide followed by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. The number of new peptide bands appearing in the gel is a function of the number of tryptophan residues, and the relative migration of the bands permits calculation of peptide molecular weights and an estimation of the positions of the tryptophan residues in the peptide chain. The technique uses a sample of about 0.5 mg and is suitable for any protein that contains a small number of tryptophan residues. These are the very specimens that are difficult to assay accurately for tryptophan by spectrophotometric or colorimetric methods. Tryptophan residues which are within about 20 residues of the ends of the peptide chain or of each other would not be detected. The specificity of the cleavage with N-bromosuccinimide was ascertained by utilizing human serum albumin, which is known to have a single tryptophan residue at position 214. The technique was then applied to a comparative study of the numbers and locations of tryptophans in the serum albumins of 16 species, namely 11 mammals, three birds and two amphibians. The number of tryptophan residues were confirmed by an independent colorimetric method. All of the mammalian albumins contained a tryptophan residue near position 213. The three avian albumins examined have no tryptophan. Frog and toad albumins contained two tryptophan residues, which appear to be situated at different positions from those in mammalian albumins.


Purification and chemical characterization of human hexosaminidases A and B. N-Acetyl-beta-hexosaminidases A and B were purified to homogeneity from human placenta. In the initial step of purification, the enzymes were adsorbed on concanavalin A-Sepharose 4B and eluted from the column with alpha-methyl D-mannosides. Subsequent purification steps included DEAE-cellulose column chromatography, QAE-Sephadex [diethyl-(2-hydroxypropyl)aminoethyl-Sephadex] column chromatography, Sephadex G-200 gel filtration and preparative disc polyacrylamide-gel electrophoresis, followed by another QAE-Sephadex chromatography for the hexosaminidase A preparation, and DEAE-cellulose column chromatography, calcium phosphate gel chromatography, Sephadex G-200 gel filtration, QAE-Sephadex chromatography and CM-cellulose chromatography for the hexosaminidase B preparation. The purified preparations, particularly hexosaminidase A, had significantly higher specific enzyme activities than previously reported. The preparations moved on polyacrylamide-gel electrophoresis as single protein bands, which also stained for enzyme activity. Sedimentation-equilibrium centrifugation indicated homogenous dispersion of the enzymes, and the molecular weight was estimated as about 110000 for both enzymes. Complete amino acid and carbohydrate compositions of the two isoenzymes were determined, and, in contrast with previous suggestions, no sialic acid was found in the enzymes.


The subunits of human hexosaminidase A.

The subunits of human hexosaminidase A. Previous studies of the subunit structure of hexosaminidase gave ambiguous results, but suggested that the enzyme was composed of six equally sized subunits. Dissociation of hexosaminidase A with p-chloromercuribenzoate produces an alkylated fragment with mol.wt. approx. 50000, which is converted into hexosaminidase S by treatment with dithiothreitol. Treatment of native hexosaminidase A with sodium dodecylsulphate results in the formation of a large and a small fragment. However, although the native enzyme has a sedimentation coefficient of 5.8S, dissociation by S-carboxymethylation and maleic anhydride treatment results in subunits exhibiting a single schlieren boundary on analytical ultracentrifugation with a sedimentation coefficient of 2.18S. These results indicate that the enzyme is composed of four subunits, each with molwt. approx. 25000-27000. The mol.wt. of the native enzymes is calculated to be approx. 110000. Our data are consistent with the subunit structures of hexosaminidases A, B and S as being alpha2beta2, beta4 and alpha4 respectively.


Triacylglycerol biosynthesis in rat adipose-tissue homogenates. The optimum cofactor requirements for triacylglycerol biosynthesis in rat adipose-tissue homogenates containing mitochondrial, microsomal and cytosolic fractions were investigated. In general the optimum concentrations of cofactors for triacylglycerol biosynthesis were found to differ from those for total fatty acid esterification. The results provided further evidence for the key role of phosphatidate phosphohydrolase in the regulation of triacylglycerol biosynthesis. Albumin was included in the incubation medium to permit the use of concentrations of added fatty acids that would swamp the effects of endogenous fatty acids. The addition of albumin had little effect on the incorporation of palmitic acid and stearic acid into lipids including triacylglycerols. By contrast, a critical concentration of albumin (about 60 muM) was required before incorporation of oleic acid or linoleic acid into triacylglycerols occurred. The system was used to study the incorporation of different 1-14C-labelled fatty acids from a mixture of unesterified fatty acids [palmitic acid 30%; stearic acid 10%; oleic acid 40%; linoleic acid 20% (molar percentages)] separately into the positions 1,2 and 3 of triacyl-sn-glycerols. In general the stereo-specific distribution of the labelled fatty acids incorporated into triacylglycerols paralleled the normal distribution of fatty acids within rat adipose-tissue triacylglycerols, suggesting that the specificities of the relevant acyltrasferases have the major role in determining the positional distribution of fatty acids within triacylglycerols.


Arginase from human full-term placenta.

Arginase from human full-term placenta. Arginase was purified about 1800-fold from extracts of human full-term placenta; the enzyme appeared to be homogenous by disc electrophoresis and molecular-sieve chromatography. The mol. wt. determination by gel filtration and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis yielded a value of 70000 for the most pure and the partially purified enzyme. The human placenta arginase is a metalloenzyme with an optimum pH of 9.1. The Km for L-arginine is 27 mM. L-Ornithine and L-lysine show competitive inhibition with Ki values of 6.3 and 14 mM respectively.


The stability, polyadenylic acid content and ribonucleoprotein form of nulcear ribonucleic acid in artichoke. A nuclear preparation, containing 60-80% of the total tissue DNA and less than 0.5% of the total rRNA, was used to characterize the nuclear RNA species synthesized in cultured artichoke explants. The half-lives of the nuclear RNA species were estimated from first-order-decay analyses to be: hnRNA (heterogeneous nuclear RNA) containing poly(A), 38 min; hnRNA lacking poly(A), 37 min; 2.5 X 10(6)-mol. wt. precursor rRNA, 24 min; 1.4 X 10(6)-mol.wt. precursor rRNA, 58 min; 1.0 X 10(6)-mol.wt. precursor rRNA, 52 min. The shorter half-lives are probably overestimates, owing to the time required for equilibration of the nucleotide-precursor pools. The pathway of rRNA synthesis is considered in terms of these kinetic measurements. The rate of accumulation of cytoplasmic polydisperse RNA suggested that as much as 40% of the hnRNA may be transported to the cytoplasm. The 14-25% of the hnRNA that contained a poly(A) tract had an average molecular size of 0.7 X 10(6) daltons. The poly(A) segment was 40-200 nucleotides long, consisted of at least 95% AMP and accounted for 8-10% of the [32P]orthophosphate incorporated into the poly(A)-containing hnRNA. Ribonucleoprotein particles released from nuclei by sonication, lysis in EDTA or incubation in buffer were analysed  by sedimentation through sucrose gradients and by isopycnic centrifugation in gradients of metrizamide and CsCl. More than 50% of the hnRNA remained bound to the chromatin after each treatment. The hnRNA was always associated with protein but the densities of isolated particles suggested that the ratio of protein to RNA was lower than that reported for mammalian cells, The particles separated from chromatin were not enriched for poly(A)-containing hnRNA.


The role of the pseudo-disaccharide neamine as an intermediate in the biosynthesis of neomycin. By using wild-type and deoxystreptamine-negative mutants of Streptomyces fradiae grown in media containing [6(-3)H]glucose or [U-14C]glucose, and by subsequent hydrolysis of the labelled neomycin produced, neamines labelled with 3H in both rings I and II, but with 14C in ring I only, were prepared. A mixture of these two forms of neamine was converted by deoxystreptamine-negative Streptomyces rimosus forma paromomycinus into neomycin (not paromomycin) with a 30% yield. The3H: 14C ratio in this neomycin was the same as the measured in neamine produced by hydrolysis of the neomycin, and in unused neamine reisolated from the incubation medium. The 3H:14C ratio in the neomycin was not affected by the presence of unlabelled deoxystreptamine during the incubation. The radioactivity in the neomycin was associated with rings I and II only. It is concluded that the added neamine is incorporated into antibiotic intact, without initial hydrolysis, and that the probable first step in the subunit assembly of neomycin is the formation of neamine.


The effects of deoxyribonucleic acid secondary structure on tertiary structure. The secondary structure of supercoiled DNA was varied by changes in ionic strength. For I = 0.075-0.4 the structure remained in the previously established branched form with only minor alterations in molecular dimensions. In 4M-NaCl, which induces linear DNA to change its secondary structure to the C structure and brings about an increase in the superhelix density of the molecule, no extra branches were observed on the molecules. The limiting factors that dictate supercoil structure seem to be the number and position of potential branch points and the proximity with which the two intertwining DNA strands can approach each other on the arms of the branches. This value is close to 10nm under the conditions described, and is 14-15nm at I = 0.2. It is suggested that such values should be borne in mind when models of chromosome structure are being constructed.


The amino acid sequence of rabbit cardiac troponin I. The complete amino acid sequence of troponin I from rabbit cardiac muscle was determined by the isolation of four unique CNBr fragments, together with overlapping tryptic peptides containing radioactive methionine residues. Overlap data for residues 35-36, 93-94 and 140-145 are incomplete, the sequence at these positions being based on homology with the sequence of the fast-skeletal-muscle protein. Cardiac troponin I is a single polypeptide chain of 206 residues with mol.wt. 23550 and an extinction coefficient, E 1%,1cm/280, of 4.37. The protein has a net positive charge of 14 and is thus somewhat more basic than troponin I from fast-skeletal muscle. Comparison of the sequences of troponin I from cardiac and fast skeletal muscle show that the cardiac protein has 26 extra residues at the N-terminus which account for the larger size of the protein. In the remainder of sequence there is a considerable degree of homology, this being greater in the C-terminal two-thirds of the molecule. The region in the cardiac protein corresponding to the peptide with inhibitory activity from the fast-skeletal-muscle protein is very similar and it seems unlikely that this is the cause of the difference in inhibitory activity between the two proteins. The region responsible for binding troponin C, however, possesses a lower degree of homology. Detailed evidence on which the sequence is based has been deposited as Supplementary Publication SUP 50072 (20 pages), at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7QB, U.K., from whom copies may be obtained on the terms given in Biochem. J. (1976) 153, 5.


Purification and properties of bovine pituitary follitropin. A reproducible procedure was developed for the purification of follitropin from frozen bovine pituitary glands. The method involved precipitation with (NH4)2SO4 and acetone, followed by ion-exchange column chromatography on CM-cellulose and DEAE-cellulose and gel filtration on Sephadex G-100. A specific radioligand-receptor assay for follitropin was used to locate the activity in eluates after column chromatography and gel filtration. The potency of the highly purified bovine follitropin as measured by Steelman-Pohley bioassay was 164 times that of NIH-FSH-S1 standard preparation. They yield of bovine follitropin was 2.9 mg/kg of frozen pituitary glands. Electrophoretically, bovine follitropin was more acidic in nature and migrated further towards the anode than lutropin and thyrotropin. The elution volume of bovine follitropin by gel filtration on Sephadex G-100 was very similar to that of bovine lutropin. The amino acid composition of bovine follitropin was similar to that of sheep and human follitropin, being rich in lysine, aspartic acid, threonine, serine, glutamic acid and half-cystine.


A new method of quantitative affinity chromatography and its application to the study of myosin. A new method of quantifying the interactions between two or three components of an interacting system, one of which is insoluble, is described. The method differs from those previously applied to affinity chromatography systems in that it does not require that elution volumes be measured, but is instead dependent on measurements of the quantity of affinity-bound material. Theoretical expressions are derived for systems in which the acceptor is immobilized. Examples presented to illustrate the validity of the theory are of the latter type and are from studies on the myosin-adenosine nucleotide-PPi system. With Sepharose-myosin columns (myosin covalently coupled to CNBr-activated Sepharose) a dissociation constant of 1.8 muM for ATP4- was found. Data were also obtained under conditions that closely approximate to those found in vivo, i.e. on columns packed with a slurry of Sephadex G-50 and precipitated myosin filaments formed at low ionic strength. The binding of MgATP2-, MgADP-, ATP4- and MgPPi2- to "filamentous" myosin in both two- (myosin and nucleotide) and three- (myosin, nucleotide and PPi) component systems at different temperatures was studied and the dissociation constants obtained agreed well with previously published values. Except for the binding of ATP4- to filamentous myosin at 4 degrees  when 85% of the protein was interacting with the nucleotide, much lower values for the number of available sites occupied by the nucleotides were as a routine found in this system. Although this apparent discrepancy is difficult to explain, it is not an anomaly of the theoretical approach and may reflect the present state of understanding of the myosin system.


Characterization of the glucose 6-phosphate dehydrogenase activity in rat liver mitochondria. Glucose 6-phosphate dehydrogenase activity in rat liver mitochondria can be released by detergent. The released activity is separated by chromatography into two peaks. One peak has the kinetic behaviour and mobility similar to the soluble sex-linked enzyme, whereas the other peak is similar to the microsomal hexose 6-phosphate dehydrogenase. There is no evidence for the existence of a new glucose 6-phosphate dehydrogenase activity in rat liver mitochondria.


Human liver glycogen phosphorylase. Kinetic properties and assay in biopsy specimens. 1. The two forms of glycogen phosphorylase were purified from human liver, and some kinetic properties were examined in the direction of glycogen synthesis. The b form has a limited catalytic capacity, resembling that of the rabbit liver enzyme. It is characterized by a low affinity for glucose 1-phosphate, which is unaffected by AMP, and a low V, which becomes equal to that of the a form in the presence of the nucleotide. Lyotropic anions stimulate phosphorylase b and inhibit phosphorylase a by modifying the affinity for glucose 1-phosphate. Both enzyme forms are easily saturated with glycogen. 2. These kinetic properties have allowed us to design a simple assay method for total (a + b) phosphorylase in human liver. It requires only 0.5 mg of tissue, and its average efficiency is 90% when the enzyme is predominantly in the b form. 3. The assay of total phosphorylase allows the unequivocal diagnosis of hepatic glycogen-storage disease caused by phosphorylase deficiency. One patient with a complete deficiency is reported. 4. The assay of human liver phosphorylase a is based on the preferential inhibition of the b form by caffeine. The a form displays the same activity when measured by either of the two assays.


Human liver alkaline phosphatase purified by affinity chromatography, ultracentrifugation and polyacrylamide-gel electrophoresis. A method is presented for the preparation of human liver alkaline phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.1). The method gives a purification factor of 12.5 X 10(3) over the initial aq. butan-1-ol extract, a recovery of 6.0% and a specific activity for the preparation of 1450-1550 units/mg of protein, 1 unit being defined as the amount of enzyme catalysing the hydrolysis of 1mumol of p-nitrophenyl phosphate/min at 35 degrees C in 0.1 M-2-amino-2-methylpropan-1-ol/HCl buffer, pH 10.5, containing 10mM-p-nitrophenyl phosphate. Homogeneity was studied by ultracentrifugation, by immunoelectrophoresis and by polyacrylamide-gel electrophoresis. A single contaminating protein was present which was less than 5% of the total. Ultracentrifugation and equilibrium-gradient-pore electrophoresis techniques indicated a mol.wt. of 156000 and 160000 respectively. Equilibrium-gradient-pore electrophoresis indicated that the alkaline phosphatase molecule is possibly a dimer, comprising two subunits of about 80000 mol.wt. Amino acid analysis proved remarkably similar to that for alkaline phosphatase from other sources, regardless of species.


Purification and properties of an alginate lyase from a marine bacterium. An unidentified pseudomonad isolated by enrichment procedures from decomposing seaweed was grown in defined medium containing sodium alginate as the sole carbon source. The alginate lyase recovered from disrupted bacterial cells was purified by a procedure of (NH4)2SO4 precipitation, gel filtration and ion-exchange chromatography. From sodium dodecyl sulphate/polyacrylamide-gel-electrophoresis experiments a mol.wt. of about 50 000 was determined. The enzyme was active against both algal and bacterial alginate preparations. Kinetic studies together with analysis of the unsaturated oligouronide products of alginate lyase action indicated the enzyme was specific for guluronic acid-containing regions of the macromolecular substrate. The specificity of the enzyme can be used to give information about the primary composition of alginate samples.


Similarities between a dipeptide hydrolase from brush-border and cytosol fractions of guinea-pig intestinal mucosa. A dipeptide hydrolase from the brush border of guinea-pig intestinal mucosa was purified. The enzyme resembles another dipeptide hydrolase isolated from the cytosol fraction of intestinal mucosa. Studies on the binding of cytosol peptide hydrolase to brush-border membranes indicate that the enzyme found in the brush border may be a cytoplasmic contaminant.


Uptake of iodinated human kidney alpha-D-mannosidase by rat liver- Association with membrane elements and stability in vivo and in vitro. 1. Human kidney alpha-D-mannosidase (form A) was labelled with 125I to a specific radio-activity of approx. 2250muCi/mg of protein, essentially without loss of enzymic activity. The enzymic activity and radioactivity of the iodinated material also co-migrated in gel filtration and gel electrophoresis. 2. The binding of 125I-labelled mannosidase in vitro to particulate material in liver and kidney homogenates was of the other of 2 pg/mg of particulate material in liver and kidney homogenates was of the order of 2pg/mg of particulate protein withing 16h at 37 degrees C, and essentially zero in intervals of up to 60 min. The degradation in vitro of labelled exogenous mannosidase was of the order of 10-20pg/ 16th per mg of protein in postnuclear supernatant, and it was saturated entirely within 1h at 37 degrees C. 3. The binding of labelled mannosidase in vivo to particulate elements of liver homogenates 60 min after intravenous injection was at least 10 times higher in terms of specific radioactivity than the highest value attainable in vitro. Virtually all exogenous enzyme bound to liver particulate material could be recovered in macromolecular form after disruption of membranes by detergents. 4. The radioactive enzyme bound to liver particulate material could be detached almost completely by shearing, repeated freezing and thawing, and exposure to strong detergents under conditions that do not eliminate rough-endoplasmic-membrane structure. It could bot be released, however, by high salt concentration (0.5M-KC1) or by exposure to weak detergents such as Tween 80. The particle-bound enzyme should thus be associated with plasma membranes and lysosome-like elements. 5. Of the rat tissues studied, only liver could approach, within 60 min after the injection, the concentration of exogenous mannosidase found in the blood serum. The activity per g tissue weight fell progressively from liver (60% of serum value) to kidney (16% of serum value), lung (8% of serum vlaue), spleen (6% of serum value) and brain (0.9% of serum value). Most of the radioactive enzyme found in tissues other than liver appeared to be present in a free form, whereas in liver more than 50% of the labelled enzyme was associated with membrane elements.


An electrophoretic study of the reversible binding of phosphate to ovalbumin. An electrophoretic method is described for the measurement of relatively weak interaction of ions  with proteins, and illustrated with the ovalbumin-phosphate system in 0.1I buffers, pH6.1. Results indicate that under these conditions the interaction of a single dibasic phosphate ion with ovalbumin is described by an association constant of approx. 250M-1.


The instantaneous monitoring of polyacrylamide gels during electrophoresis. The advantages of being able to see protein zones in a gel during electrophoresis (and hence before staining) are pointed out, and a method is described which depends on local increments of refractive index in these zones. The use of local increments of refractive index in polyacrylamide gels for measuring protein concentrations in zones during electrophoresis is briefly considered; it is found that such increments are greater than would be expected from the amount of protein when sodium dodecyl sulphate is present. The enhancement depends on conditions and time of running. This makes quantitative estimates difficult, but the sensitivity of detection of protein zones by observations based on refractive-index changes is greatly increased by this property of sodium dodecyl sulphate. Methods are described for making optically uniform gels (both with uniform and with graded concentrations of polyacrylamide), necessary for observation of small changes in refractive index. A simple dark-field system of observation is described. Examples are given showing protein samples observed with the system during electrophoresis and compared with the same gel stained with Coomassie Blue after completion of the run. Under optimal conditions the optical method is comparable in sensitivity with staining. With the proteins of lower mol.wt. (approx. 15000), the optical method is not so sensitive, becoming less sensitive with longer running time. This loss of sensitivity is greatly decreased by using more concentrated polyacrylamide gels, and graded gels are therefore more suitable for optical observation than are uniform gels. The observation of protein zones during electrophoresis adds nothing to the time needed for making a stained gel and gives much information long before it can be obtained from the stained gel.


The purification and characterization of rabbit placental lactogen. Rabbit placental lactogen, a polypeptide hormone functionally related to the growth hormone/prolactin family, was isolated from placenta by (NH4)2SO4 precipitation, gel filtration and ion-exchange chromatography on DEAE-and CM-cellulose. The hormone was purified to more than 90% homogeneity, as determined by end-group analysis. On disc gel electrophoresis at pH9.0 it migrates as a pair of closely spaced bands with mobilities of 0.489 (minor band) and 0.511 (major band), and its isoelectric point is 6.1. Its mol.wt. is 20600, as determined by sedimentation--equilibrium centrifugation, and 24200, as estimated by gel electrophoresis in sodium dodecyl sulphate. Its amino acid composition resembles that of rabbit growth hormone and rat prolactin, except for a lower glutamic acid and leucine content. Like the prolactins, rabbit placental lactogen has two tryptophan and six cysteine residues, and its N-terminus, valine, is identical with that for human placental lactogen. By radioimmunoassay, it does not cross-react with antisera to either rat growth hormone or rat prolactin; in addition, it does not cross-react with antisera to bovine placental lactogen by double immunodiffusion. The similarity of the biochemical characteristics of rabbit placental lactogen to the other non-primate placental lactogens lends further support to the hypothesis that these molecules occupy a more central position in the growth hormone/prolactin "tree" than do their primate counterparts.


Aldolase binding to actin-containing filaments. Formation of paracrystals. Electron-microscopy observation show that when aldolase binds to F-actin or F-actin-tropomyosin, highly ordered paracrystalline structures are formed consisting of tightly packed filament bundles cross-banded at 36 nm intervals. Morphologically different paracrystalline arrays are formed between aldolase and F-actin-tropomyosin-troponin. The filament bundles are far more extensive and are characterized by a prominent cross-striation at 38nm intervals. It is suggested that this reflects an interaction between troponin and aldolase.


Chemical synthesis of all-trans-beta-retinoyl phosphat. all-trans-beta-Retinoic acid is phosphorylated to retinoyl phosphate by bis(triethylamine) phosphate with yields of 10-15%. The product is soluble in methanol and is eluted from DEAE-cellulose acetate at a concentration of 0.1M-ammonium acetate in 99% (v/v) methanol. Its phosphate/retinoic acid molar ratio is 1. Retinoyl phosphate has an absorption maximum at 360nm in methanol, whereas retinoic acid has a maximum at 350 nm. The compound is hydrolysed at pH2 and pH13 for 20 min at 37 degrees C, but is relatively stable under the same conditions at pH4, 6, 8 and 10. Retinoyl phosphate (RF 0.1) can be separated from retinyl phosphate (RF 0.2) by chromatography on thin layers of silica gel in chloroform/methanol/water (60:25:4, by vol.).


Conversion of (+/-)-synephrine into p-hydroxyphenylacetaldehyde by Arthrobacter synephrinum. A novel enzymic reaction. A partically purified enzyme from Arthrobacter synephrinum was found to catalyse the conversion of (+/-)-synphrine into p-hydroxyphrenylacetaldehyde and methylamine. The enzyme is highly specific for synephrine and is  distinctly different from monoamine oxidase.


Metabolism of daunorubicin by a barbiturate-sensitive aldehyde reductase from rat liver. A barbiturate-sensitive aldehyde reductase was purified to homogeneity from rat liver and shown to metabolize the cancer-chemotherapeutic antibiotic daunorubicin. The aldehyde reductase may have important roles in the metabolism of exogeneous drugs as well as the aldehyde derivatives of the biogenic amines.


The relationship between adrogen receptors and the hormonally controlled responses of rat ventral prostate. 1. The administration of dihydrotestosterone to rats orchidectomized 7 days previously stimulated the synthesis of nuclear receptor in prostatic cells several hours in advance of DNA synthesis and mitosis. 2. The synthesis of nuclear receptor is tightly coupled to cell proliferation; consequently, in resting cells, there is no further net synthesis of nuclear receptor above the maximum of approx. 8000 molecules/cell. 3. After orchidectomy a rapid decline in the concentration of free androgen in the nuceus and a slower decline in the concentration of nuclear receptor are observed. 4. Owing  to the apparent scarcity of receptor-inactivating factors in the nucleus, and the inverse relationship between amounts of nuclear and cytoplasmic receptors, it is concluded that the nuclear receptor is discharged into the cytoplasm after orchidectomy. 5. The formation of the cytoplasmic receptor is an early event preceding the onset of cellular autolysis. 6. Regressing prostate develops the capacity to eliminate cytoplasmic receptor, and this capacity is retained by the regenerating prostate for at least 14 days. 7. The synthesis of nuclear receptor in early G1 phase may control the entry of cells into the cell cycle and the prolonged retention of receptor in the nucleus may prevent the activation of autophagic processes.


Isolation of pure cholinergic nerve endings from the electric organ of Torpedo marmorata. A rapid method for the preparation of highly purified cholinergic nerve endings from the electric organ of Torpedo is described. The endings retain their cytoplasmic components, as shown by biochemical and morphological observations. The homogeneity of these synaptosomes make them a useful tool for further studies.


Collagen synthesis and deposition in cartilage during disrupted proteoglycan production. A simple system was developed to investigate in vitro the possible relationship between collagen and proteoglycan synthesis in cartilage. When production of complete proteoglycan molecules was effectively inhibited with 4-methylumbelliferyl beta-D-xyloside collagen synthesis and distribution were virtually unaffected.


Arteriovenous differences for amino acids and lactate across kidneys of normal and acidotic rats. 1. Arteriovenous differences fro amino acids across kidneys of normal and chronically acidotic rats were measured. Glutamine was the only amino acid extracted in increased amounts in acidosis. There was a considerable production of serine by kidneys from both normal and acidotic rats. 2. The arterial blood concentration of glutamine was significantly decreased in acidotic animals. 3. The glutamine extracted by kidneys of acidotic rats was largely and probably exclusively derived from the plasma. 4. The blood lactate concentration was unchanged in acidosis, as was the uptake of lactate by the kidney.


Resistance to hepatic action of vasopressin in genetically obese (ob/ob) mice. 1. Fatty acid synthesis, measured in the perfused liver of genetically obese (ob/ob) mice with 3H2O or [14C]actate, did not show the inhibition by [8-arginine]vasopressin (antidiuretic hormone) that is observed in livers from normal mice. 2. Hepatic glycogen breakdown in obese mice was stimuulated by vasopressin, but not as extensively as in lean mice. 3. If obese mice received a restricted amount of food, then fatty acid synthesis still did not respond to vasopressin, but glycogen breakdown was fully stimulated. 4. Cholesterol synthesis was not inhibited by vasopressin in livers from obese mice. 5. Vasopressin inhibited fatty acid synthesis in intact lean mice, but not in obese animals. 6. These results suggest that genetic obesity could be due to an inborn error within the mechanisms (other than adenylate cyclase) which mediate responses to extracellular effectors.


Stimulation of collagen galactosyltransferase and glucosyltransferase activities by lysophosphatidylcholine. Lysophosphatidylcholine stimulated the activities of collagen galactosyl- and glucosyl-transferases in chick-embryo extract and its particulate fractions in vitro, whereas essentially no stimulation was noted in the high-speed supernatant, where the enzymes are soluble and membrane-free. The stimulatory effect of lysophosphatidylcholine was masked by 0.1% Triton X-100. In kinetic experiments lysophosphatidylcholine raised the maximum velocities with respect to the substrates and co-substrates, whereas no changes were observed in the apparant Km values. Phospholipase A preincubation of the chick-embryo extract resulted in stimulation of both transferase activities, probably gy generating lysophosphatides from endogenous phospholipids. No stimulation by lysophosphatidylcholine was found when tested with 500-fold-purified glycosyltransferase. The results suggest that collagen glycosyltransferases must be associated with the membrane structures of the cell in order to be stimulated by lysophosphatidylcholine. Lysophosphatidylcholine could have some regulatory significance in vivo, since its concentration in the cell is comparable with that which produced marked stimulation in vitro.


Testosterone control of nucleic acid content and proliferation of epithelium and stroma in rat seminal vesicles. Tissue wet weight, nucleic acid content and epithelial and stromal cell numbers were measured in the seminal vesicles of sexually mature male rats. After castration, tissue weight and RNA decreased rapidly and in aprallel to reach, after 14 days, values only 15-20% of those in control (not castrated) animals. During this period, DNA decreased to a much lesser extent (by about 40%), but this change in DNA correlates well with the observed loss of cells from the epithelium. Testosterone in vivo promoted an immediate resynthesis of RNA, the value characteristic of control animals being reached within 80h. Delays occurred in the hormone-induced regain of tissue weight (30h) and DNA (40h), each of which preceded proliferation of the epithelium (40--50h). The cells of the stroma were unaffected by these changes in the androgenic statls of the animal. It is suggested that these proliferative changes in the epithelium cannot account for the previously reported induction by testosterone of basic secretory proteins in this tissue.