Experience of changing pyrogenicity in albumin solutions.

A number of production lots of an intravenous preparation of 4.3 g/dl of human albumin failed to EP pyrogen test when tested in three rabbits. Several lots were reprocessed and subsequently proved satisfactory but over thirty lots were placed in storage and monitored by both the EP test and by using the LAL test over a period up to 650 days. The majority of the lots showed a steady fall in the pyrogen response by both tests. These lots were removed from storage and reprocessed by sterile filtration and dispensing into new glass containers. Further examination of the product showed it to be satisfactory in all regards. The product of several lots was used in treatment episodes involving the use of cell separators and shown to be satisfactory in clinical use. The remaining lots were also issues for general clinical use without incident.


Limulus amebocyte lysate testing of nomral serum albumin (human) in the United States since 1975.

In order to determine the reactivity of Limulus Amebocyte Lysate to albumin products, over 1500 lots of 25% Normal Serum (Human) produced by 12 U.S. licensed manufacturers were tested over the past 4 years. Representative samples of 5% Normal Serum Albumin (Human) and Purified Protein Fraction (human) were also tested with Limulus Amebocyte Lysate. Problems associated with apparent false negative reactions are discussed. Model test protocols are presented that might be useful in determining the degree of false positivity of albumin products. These models could be adopted by blood fractionators to gather validation data to support substitution of the Limulus Amebocyte Lysate test for the rabbit pyrogen test.


Calibration of the limulus test for blood products.

The Protein Fractionation Centre has been using the Limulus Amoebocyte Lysate test for detecting the presence of pyrogens for some five years. All tests on final plasma products have been carried out in parallel with the EP rabbit pyrogen test. Good correlation between both tests has been obtained with products such as distilled water and crystalloid solutions. However, it is more difficult to correlate results when dealing with plasma proteins. Each product appears to react differently. Some normally pass the limulus test undiluted while others, such as immune globulins and albuminoid products, have to be diluted to various concentrations before they pass. A preliminary statistical analysis has been carried out on the results of pyrogen testing on Stable Plasma Protein Solution, firstly to ascertain that a significant correlation does exist between the LAL test and the rabbit test, and secondly to try to obtain "cut off" values for the limulus test which compare well with the official EP temperature rises in the rabbit test.


Improved methodology for the quantitative analysis of mixtures by band centrifugation: reversible equilibration in immunoglobulin preparations.

Double-sector aluminium centerpieces are modified for use in analytical high-speed band centrifugation up to 60,000 rpm. A method has been developed for testing IG preparations according to administrative regulations with special regard to precision and reproducibility. The proposed band-forming centrifugation method with scanner registration in UV absorption optics and special evaluation of the curves gives values in comparison to well-known procedures by Svedberg and Pedersen. Subjective and systematic errors are especially estimated with this method and the sensitivity is best suited to automatic computerization and results are therefore readily available at the end of the centrifuge run within a few minutes. Investigations have been undertaken to analyze reversible equilibration in immunoglobulin preparations; the results are discussed.


Control of blood group specific substances A and B and human chorionic gonadotropin in human immunoglobulins and albumins.

One hundred and thirty-six lots of normal human immunoglobulins and 95 lots of human albumins were tested for blood group substances A and B using haemagglutination inhibition assay. Blood group substances A and B were revealed in all the samples of commercial preparations from placental origin. The haemagglutination inhibition test is very suitable for routine control but since the only standard is the U.S. standard for blood group substances A and B, an international standard for these substances is urgently needed. Sixty-five lots of normal human immunoglobulins and 15 lots of human albumins were assayed against the Second International Standard for Human Chorionic Gonadotropin (HCG) using three biological and three immunological methods. Hormonal activity was revealed in all the preparations from placental origin. The results obtained by immunological methods were found to be considerably higher than those obtained by biological methods which indicate that the activities measured by the two groups of methods were different. The control of preparations is discussed. Antibodies to HCG were found in the immunoglobulin preparations as well.


Chemical contaminants in human serum albumin: presence of glucose and ethanol in freeze-dried or ultrafiltered products.

Glucose and Ethanol are determined in final Human Serum Albumin solutions, obtained by two different processes: freeze-drying (FD) and ultrafiltration (UF). The elimination rate for glucose is higher than that for ethanol in case of ultrafiltration. The concentration of both "contaminants" seems to be much lower by UF than by FD. From these data arises the question of a limit-value for fractionating agents, in the final product. In the case of FD process, as the reagents remain in the product final, they all must be strongly controlled. As a large part of small molecules can be eliminated by UF process, it permits to seek new reagents for protein separation.


Gel permeation chromatography as a stability-indicating assay for human serum albumin.

It has been known for a considerable period of time that preparations of normal human serum albumin contain a amount of polymers and their formation appears to be related to the stability of human albumin solutions. This report describes a gel permeation chromatography technique using Sephadex G-150 to separate albumin molecules into monomers, imers, and polymers. In order to determine if this system could detect small changes in albumin solutions, 5% solutions of albumin were heat denatured and assayed. The changes which resulted from the heat denaturation were easily measurable. Additional experiments using 5% and 25% albumin solutions showed that this system could also detect changes in the amount of aggregation which occurred in these solutions with time. It therefore appears that this technique may be of value in assessing the stability of albumin solutions as measured by the formation of protein aggregates.


Genetic transmission of Moloney leukemia virus: mapping of the chromosomal integration site.

Mice genetically transmitting the exogenous Moloney leukemia virus (Balb/Mo) have been previously derived. These animals carried one copy of Moloney virus DNA (M-MuLV) in their germ line and transmitted the virus as a single Mendelian gene to the next generation. Homozygous BALB/Mo mice were used to genetically map the M-MuLV locus. Embryo fibroblasts were fused to established Chinese hamster cells and somatic cell hybrids were selected. Segregation of mouse chromosomal markers in the hybrids was correlated to the loss of M-MuLV-specific sequences as detected by molecular hybridization. Of 15 isozymes located on different mouse chromosomes only triosephosphate isomerase segregated syntenic with the M-MuLV gene, suggesting that the virus was integrated on chromosome No. 6. This was confirmed by sexual genetic experiments analyzing segregation of Moloney viremia and two markers on chromosome 6 and 15, respectively. The results show that M-MuLV expression is linked to wa-1 on chromosome 6 at a distance of about 30 map units. These data define a new genetic locus, Mov-1, representing the structural gene of M-MuLV in BALB/Mo mice.


Integrated interpretation of fetal heart rate, intrauterine pressure and fetal transcutaneous PO2.

By a study of the patterns of changes and the temporal relationship in fetal heart rate, intrauterine pressure, fetal tcPO2 and 'flow', a series of ten typical examples of combined patterns are described. Fetal tcPO2 is affected by the level of the fetal oxygenation and by the blood flow beneath the electrode. Should the latter be below a certain critical level fetal tcPO2 will be lower than fetal scalp blood PO2. By the integrated analysis of the four simultaneously recorded variables the different factors dominating fetal tcPO2 may be identified.


Chronic occlusion of the rabbit Fallopian tube with silicone polymer.

Investigators have previously reported good retention, antifertility efficacy, and good retrievability with no adverse tissue reactions in rabbit oviducts occluded with silicone rubber for 55 days. The present study attempts to determine the long-term effects of this silicone polymer system installed into the oviducts of 12 New Zealand White rabbits for 7 months. 9 rabbits had both oviducts occluded while 3 had only one oviduct treated. All animals were visualized by X-rays and were mated an average of six times over the experimental period. Oviduct tissue was examined with light and scanning electron microscopy. There was no evidence of inflammation, distortion, or tissue irritation. Good retention and antifertility efficacy were exhibited.


Diagnostic aid of endometrium biopsy.

The 784 cases, from which 701 complained of abnormal uterine bleeding, were reviewed by age groups, presenting symptoms, sex steroid medication and endometrial findings. Endometrial polyps and endometritis, which showed an incidence of 17.5 and 10.0%, respectively, were correlated with clinicopathologic findings. Peri- and postmenopausal bleeders were investigated with special reference to hormone therapy and endometrial patterns. Dysfunctional uterine bleeding was diagnosed in 48.7% of the subjects during reproductive years. Pathologic findings were found to explain the clinical symptoms in 60.6% of the entire study group.


Evidence of lack of passage of human prolactin through human placental membranes.

Human prolactin (hPRL) derived from either amniotic fluid (AF) or pituitary was tested for its passage capability through amniotic and chorionic membranes placed in an in vitro perfusion chamber. The passage of both 'cold' and labelled prolactin were investigated using intact monomeric iodinated hPRL isolated from amniotic fluid which also served as a tracer in a homologous double antibody radioimmunoassay used for prolactin determinations. Throughout 4 h incubation period at 37 degrees C, there was no passage of 'cold' hPRL from either of the sources across either direction. Also, in contrast to free iodine ions, there was no passage of intact labelled prolactin in either direction. Decomposition of labelled prolactin occurred only in the chorionic side, and only decomposed labelled prolactin molecules passed into the amniotic side. These became detectable 1 h after commencement of incubation. These results may explain the long half-life and the high concentration of hPRL in amniotic fluid, and also suggest that the source of AF hPRL is not maternal.


Ixodid ticks of Barun Glacier region (the Nepal Himalaya).

During the 3rd Czechoslovak mountaineering expedition to the Himalaya in the spring 1973 ticks of the family Ixodidae were collected from small mammals and additionally from pastured sheep, and humans. The tick material represented the following species: Ixodes (P.) ovatus Neumann, 1899, Ixodes (I.) acutitarsus (Karsch, 1880) and Haemaphysalis (A.) aponommoides Warburton, 1921. Ecological and zoogeographical characteristics of the ticks found are given and their potential medical importance is discussed.


Allergogenous mites (Acari: Pyroglyphidae) in private recreation houses.

The bed fauna of periodically inhabited dwellings markedly differs from the bed fauna in town apartments, both in quantity--a higher number of mites and quality--an important dominance of the species Euroglyphus maynei which makes up 70.4% of mites in country cottages, but only 28% in town apratments used by the owners of such recreation houses. In other apartments it is very rare. There differences are explained by the specific regime of humidity and temperature in the periodically inhabited rooms. The patients' worsening condition is apparently caused by quantitative changes.


Some ecological criteria of natural focality of mycotic zoonoses.

Mycotic zoonoses sensu stricto and mycotic infections acquired from animal polluted environment belong to diseases characterized by the phenomenon of natural focality. On a global scale they comprise 16 different diseases caused by yeast-like organisms, dematophytes, dimorphous agents of systemic mycoses and primarily saprophytic fungi as well. Prevalence of mentioned diseases is influenced by a number of factors concerning not only biology and ecology of pathogenic agents and particular hosts, but also character of specific extra-animal substrates which make possible long-term maintenance or active propagation of agents in the environment. On the basis of their experience and published data as well, the authors present a survey and analysis of these factors, with particular emphasis on very frequent dermatophytozoonoses and some systemic mycoses. They pay attention to different conditions in urbanized and rural areas and to specific situations encountered by man and food producing animals in these areas. They note different ways of heterotrophy of particular agents and their association with vertebrate hosts which they divide into six ecologically different groups. The authors also characterize the environment in which a portion of saproparatrophic circulation of the agent takes place. They also give a survey of animal-connected human mycoses, which may arise due to occupational hazards.


Rickettsiae of the spotted fever group in Hungary.

Rickettsiae were detected by haemocyte test in 7.2% of Dermacentor marginatus and 4.7% of D. reticulatus ticks collected in Hungary. Six strains of rickettsiae were isolated from positive specimens. These rickettsiae according to CFR are closely related, probably identical with R. slovaca and they represent the first rickettsiae of the Spotted Fever group found in that country.


[Influence of carrageenin inflammation on prostaglandin release from rat synovial membrane and action of anti-inflammatory drugs (author's transl)].

Carrageenin was injected into the knee joint cavity of rats, and influence of anti-inflammatory drugs on prostaglandin (PG) release in vitro from inflamed synovial membrane was studied. Release of PGE and F from inflamed synovial membrane was increased. Aspirin suppressed PGE and F release from non-inflamed and inflamed synovial membrane in vivo and in vitro, but suppression of PGF release was more intense. Hydrocortisone suppressed release of PGE and F from inflamed synovial membrane in vivo, and did not influence PG release in vitro. Release of PGE and F fron non-inflamed synovial membrane was not influenced by hydrocortisone in vivo and in vitro. PGE content in inflamed synovial membrane was increased markedly, and PGE and F content in inflamed and non-inflamed synovial membrane was decreased markedly by aspirin in vivo. Content of PGE and F in non-inflamed and inflamed synovial membrane showed a tendency to decrease with hydrocortisone administration in vivo, but such was not significant.


[Effect of spasmolytics on cathartic activity of pilocarpine, 5-HT and prostaglandlin E2 in mice (author's transl)].

Effects of autonomic, anti-histaminic, anti-5HT drugs, papaverine and morphine on cathartic activity of spasmogens such as pilocarpine, 5-HT and prostaglandin E2 (PGE2) were tested by giving these drugs to mice subcutaneously. Mice used were male dd strain weighing 20 +/- 2 g and the catharatic effect was evaluated by the all or none method. The cathartic effect by pilocarpine was inhibited markedly by anti-cholinergic drugs such as atropine and diphenhydramine, but not by hexamethonium and methysergide. Therefore, the action of pilocarpine was found to be direct on the acetylcholine receptors. The catharatic effect by 5-HT was inhibited significantly by hexamethonium and methysergide. Atropine inhibited the cathartic effect of 5-HT, but the inhibitory activity was lower by about one hundredth than that on pilocarpine-induced diarrhea. It is suggested that 5-HT has dual sites of actions and there are less cholinergic and specific serotonergic actions. The cathartic effect by PGE2 was inhibited by anticholinergic, anti-5HT and ganglion blocking agents. PGE2 appears to possess the same mode of action as both pilocarpine and 5-HT. Papaverine inhibited little the cathartic effect of all three spasmogens, while morphine had a potent and nonspecific inhibitory effect on the cathartic action of all three spasmogens.


[A fundamental study on a bioassay for the antiphlogistic effect of topically applied antiinflammatory agents (author's transl)].

To establish a reliable laboratory assay for quantitating topical anti-inflammatory efficacy, the method of Tonelli et al was modified by employing felt fixed forceps, felt and sharp punch in Wistar rats. Croton oil, applied topically to the rat ear, elicited an acute phlogistic response which was maximal 6 hr after the application. The phlogistic response elicited by a single topical application of croton oil (1, 2, 5 and 10%) was increased in a dose-dependent manner, and croton oil, 5%, induced 63.2% increase of ear weight and was the optimal concentration for the experiment. Using this procedure, the antiphlogistic potencies of two corticoids were assayed under conditions of a blind test. ED50 of betamethasone valerate and diflucortolone valerate was 0.26 mg/ml and 0.0097 mg/ml, respectively, in Wistar rats, and 0.86 mg/ml and 0.016 mg/ml in Sprague Dawley rats, suggesting that diflucortolone valerate has an antiphlogistic potency of 27-56 relative to that seen with betamethasone valerate with minor differences in the strain of the rat. Our method should prove to be a useful assay for rapidly quantitating antiphlogistic activities of topically applied corticoids.


[Irritative activity of a new anti-inflammatory agent 4-(p-chorophenyl)-2-phenyl-5-thiazoleacetic acid (CH-800) on the gastrointestinal tract in rats (author's transl)].

A single oral administration of CH-800 induced a dose-dependent irritation of the stomach and intestine. As determined from the UD50 value (the dose inducing ulceration by 50%), the potency of gastric irritation was as follows; indomethacin greater than diclofenac Na greater than ibuprofen greater than aspirin greater than CH-800 greater than phenylbutazone. Repeated administrations of CH-800 for 5 days induced a gastric irritation when given in doses from 3 to 100 mg/kg, however, the response was not dose-related. In contrast, the irritation of intestinal mucosa seen with CH-800 administration was dose-related. The degree of gastric or intestinal irritation seen with dosing of other drugs was as follows; indomethacin greater than diclofenac Na greater than ibuprofen greater than aspirin greater than phenylbutazone or indomethacin greater than CH-800 = diclofenac Na greater than ibuprofen greater than phenylbutazone, respectively. CH-800 given for 5 days significantly delayed the healing of active ulcers and the healed ulcers showed a tendency toward re-ulceration. However, the irritating activity of phenylbutazone, diclofenac Na and ibuprofen was more potent than that of CH-800. Thus, CH-800 appears to have a rather weak irritative activity on the gastrointestinal tract of rats without ulceration, in contrast to other commonly clinically prescribed drugs.


[Etiology of membranous nephropathy in children: Association between membranous nephropathy and hepatitis B virus infection (author's transl)].

A total of 163 Japanese children (90 boys and 73 girls, ranging in age from 3 to 15) with proteinuria and/or hematuria were studied for renal histopathology using biopsy materials by light microscopy, electron microscopy, and immunofluolescence method. Eleven patients were diagnosed as membranous nephropathy (MN) while the other 152 patients had various renal diseases other than MN. All patients with MN did not have any known predisposing of associated caused. Hepatitis B virus surface antigen (HBsAg) in the serum, as determined by a reversed passive hemagglutination method (RPHA), was positive in 100% of the patients with MN, while it was positive in 4.6% of the patients with other renal diseases. The difference was statistically significant. Of the 11 mothers of the children with MN, six were positive for HBsAg, and one of the remaining 2 was positive for antibody to HGsAg (anti-HBs) and another was not examined. These findings suggest that MN among Japanese children are mainly, if not exclusively, caused by hepatitis B virus (HBV) and that the virus is transmitted from the mother to child in most instances. In each case of HBsAg-associated glomerulonephritis reported, HBsAg was detected, by immunofluorescence, in the glomeruli, with a distribution similar to that of immunoglobulins. However deposits of HBsAg could not be demonstrated in the glomeruli of the 9 patients with MN studied. Pathogenic immune complexes in the glomerular lesions with subepithelial deposits have been shown to weigh less than 1 million daltons. Since the intact 20-nm HBsAg has a molecular weight of more than 2.4 million daltons, probably most immune complexes containing it would be very large and rapidly cleared by the reticuloendothelial system. Therefore, this failure to detect glomerular staining with anti-HBs antiserum may mean that MN is caused by some other antigen, of a lower molecular weight, associated with HBV, but not necessarily by HB surface antigen.


[Experimental and clinical studies on the mechanism of alcoholic fatty liver (author's transl)].

In order to clarify the mechanism of alcoholic fatty liver, rats were reared with alcohol diet from adult, foetal or weanling periods. When rats were fed with the diet from adult for 4 weeks, these livers showed apparent fatty deposition histologically and biochemically. Low density lipoprotein (LDL) in serums of these rats lowered in level significantly than control, which indicate decreased production of secretion of very low density lipoprotein (VLDL) in the liver from which converted to LDL in peripheral tissues. When rats were reared with the diet from their foetal life, their livers deposited little of fat at any time examined after birth up to 13 weeks in spite of good diet uptake. The same phenomenon was observed in rats reared from their weanling period with the diet. In situ liver perfusion was performed to clarify the anti-fatty liver mechanism. Ketone body (KB) production rates from palmitate infused constantly at physiological concentration and bile production rates were not different among control, weanling and foetal groups. Oxygen consumption rate in control decreased after infusion of palmitate-albumin complex solution. However the rates in weanling and foetal alcohol rats increased significantly after infusion of the solution. The latter group showed more increase in rate than the former. When theoretical oxygen consumption for production of KB was compared with actual one, livers from control rats seemed to use the whole oxygen for KB production. On the other hand only 59.7% of whole oxygen in foetal alcohol group and 85.9% in weanling group used for KB production respectively. It is surmized that the increased non ketone oxygen in these groups, especially in foetal alcohol group, indicate the anti-fatty liver mechanism in these groups, probably augmented FFA oxidation. Electrophoretic analysis of lipoproteins in chronic alcoholisms showed decreased beta and pre-beta band and increased alpha band. These changes returned to normal after 1 week of admission. These experimental and clinical data indicate that impaired oxidation of FFA is an important factor for the formation of alcoholic fatty liver and impaired transport of fat from liver might enhance the change in the liver.


[Histological study of the paracervical ganglion (Frankenhäuser) in mice (author's transl)].

The paracervical ganglion in dd-mice ranging in age from birth to 90 days was studied qualitatively and quantitatively by light microscopy. The paracervical ganglion is located just lateral to the fornix vaginae, and is shaped like a flattened cone. Out of the parenchymal cells, nerve cells are 97% and chromaffin cells are 3%, respectively. The chromaffin cells are arranged in small clusters or individually, and are scattered throughout the ganglion. Their chromaffin reaction varies in density from cell to cell. In neonatal mice, the ganglion consists of nerve cells and small spherical cells. With advancing age, as the small cells decrease in number, chromaffin cells appear. At 20 days of age, the ganglion is almost similar in structure to that of the adult. The nuclear and cellular size distribution curves of nerve cells are symmetrical in pattern after the age of 20 days. The nuclear size distribution curves of the chromaffin cells are symmetrical and are almost the same in pattern in newborns as in adults. The cellular size distribution curve of chromaffin cells are symmetrical before 20 days of age, but, with advancing age, the curve shifts to the larger size and becomes asymmetrical in pattern due to an increase of larger cells.


[A comparative study between radiologic and macroscopic findings of small cancers less than 1 cm in the antrum of the stomach (author's transl)].

A comparative study was made of macroscopic and radiologic findings about 95 lesions (83 cases)  of small cancers (10 mm and below) in the antrum of the stomach, and the following results were obtained. 1) Preoperative diagnostic rate was 11/66 (17%) for lesions of micro-gastric cancers (5 mm and below), and 21/29 (72%) for lesions of small gastric cancers 5.1-10). Lesions of microgastric cancers which were preoperatively diagnosed as such were those with size of 3 to 5 mm accompanied by marginal elevations. 2) Radiologic findings about micro-gastric cancers. Some of the micro-gastric cancers which gave some findings in barium filled pictures existing along the greater or lesser curvature were observed as pictures of slight depression. In the compression pictures were noticed niches of irregular linear or stellar shapes with surrounding translucency. For qualitative diagnosis, the compression method was found most effective. It is considered very important to look for small niches, slight translucency, and slight deformity in the greater or lesser curvature carefully.


[Juvenile, non-progressive muscular atrophy localized in the hand and forearm. Report of 16 cases (author's transl)].

Sixteen cases of juvenile, non-progressive muscular atrophy localized in the hand and forearm were seen at our neurology out-patient clinic for the past 8 years. The analyses of these 16 cases disclosed characteristic features as follows: 1) juvenile onset 2) male preponderance 3) unique distribution of muscular atrophy and weakness in the hand and forearm. 4) insidious onset, initial progressive period and subsequent non-progressive course 5) tendon reflexes of the arms are hypoactive in half of the cases 6) no pathological reflexes 7) cold paresis 8) no definite sensory disturbance 9) no cranial nerve involvement 10) neurogenic patterns on EMG According to these features, this clinical entity carrying good prognosis must be differentiated from several diseases associated with similar muscular atrophy of extremities, especially amyotrophic lateral sclerosis which is notorious as a fetal disease.


[Influence of food browning on nutrition. III. Effect of brown substances on digestive enzyme activity (author's transl)].

In the present study, we separated and partially purified brown substances from eggplants and examined their inhibitory action on trypsin activity. The following results were obtained: 1. The first half of the elute after passing ethanol-extractable brown substances through DEAE-cellulose column showed no inhibitory action on trypsin, whereas the middle portion of the elute contained a trypsin inhibitor(s). Similar results were obtained after fractionation with Sephadex G-25. The degree of inhibition was increased after purification. 2. Both crude ethanol extracts of eggplant brown substances and acetate buffer extracts from eggplant exocarps showed similarly an enzyme inhibition of competitive type. 3. Both nondialyzable portion of ethanol extracts and purified fraction after Sephadex G-25 passage showed a noncompetitive type of inhibition. DOPA-melanin and chlorogenic acid-melanin as model substances exhibited a similar noncompetitive inhibition. Purified ethanol extracts of eggplant brown substances showed an ultraviolet absorption spectrum similar to that of chlorogenic acid. From these findings it is concluded that both eggplant brown substances and polyphenol substances play an essential role in the inhibition of digestive enzymes.


Cardiovascular response to mental stress in normal adolescents with hypertensive parents. Hemodynamics and mental stress in adolescents.

The hemodynamic response to mental stress (mental arithmetic) was studied in adolscents with varying risk factors for essential hypertension (EH), One group (genetic) consisted of normotensive well adolescents who had at least one parentnt with EH. Another group (labile) consisted of adolescents with labile hypertension each of whom also had at least one pare with EH. The control population consisted of normotensive adolescents with a negative family history of EH. Subjects with labile hypertension demonstrated a sustained increase in systolic and diastolic pressure and heart rate during stress. This response was significantly different than the control population (P less than THE CONTROL POPULATION (P LESS THAN 0.001). The stress response of the normotensive genetic population was qualitatively similar to the group with labile hypertension and significantly different than the controls in diastolic pressure and heart rate (p less than 0.001, less than 0.02). Post-stress plasma catecholamines were higher in the labile hypertensive and genetic groups than in the control group. These findings demonstrate increased central nervous system mediated adrenergic activity and cardiovascular response in labile hypertension and also in some normotensive subjects with a genetic risk for hypertension.


Cardiac output and peripheral resistance in strains of rats sensitive and resistant to NaCl hypertension.

The interrelationship of blood pressure, cardiac output, and peripheral resistance was studied in Dahl "S" and "R" rats after 3 days on a high (8%) NaCl diet. Both "S" and "R" rats were normotensive when fed a normal (0.3%) NaCl diet. After 3 days of the high NaCl diet, the "R" rats remained normotensive (BP 112 mm Hg), while the "S" rats had an elevation of arterial pressure (BP 133 mm Hg) (p less than 0.001). The cardiac outputs of both "S" and "R" rats were similar on the low NaCl diet. After 3 days of high NaCl feeding, the cardiac output of the "R" rats rose 18% above the "R" control level (p less than 0.0001), while the peripheral resistance declined 14% below the "R" control level (p less than 0.005), and the blood pressure (BP) did not change, a pattern quite contrary to the concept of "whole-body" autoregulation. With a similar 3-day high NaCl feeding in "S" rats, cardiac output (p less than 0.005) and peripheral resistance (p less than 0.05) both increased 10%, while BP rose 20%. After 7 days of high NaCl feeding, the cardiac output of the "S" rats had returned to normal, while blood pressure and peripheral resistance both continued to be elevated. This pattern of response in "S" rats could be compatible with the concept of "whole-body" autoregulation. However, since both NaCl hypertension and Goldblatt hypertension can occur in settings in which "whole-body" autoregulation appears not be to causally related, one cannot be certain whether "whole-body" autoregulation is playing a causal role in the mechanism of NaCl-induced hypertension in "S" rats. It is a striking dichotomy that 3 days of high salt feeding produces vasoconstriction in "S" rats and vasodilation in "R" rats.


Lack of correlation between serum dopamine-beta-hydroxylase activity and blood pressure in middle-aged men.

The activity of serum dopamine-beta-hydroxylase (DBH) was measured in 1194 asymptomatic middle-aged men with diastolic blood pressure ranging from 75 to 125 mm Hg during the baseline examination of a multifactorial intervention program for primary prevention of coronary heart disease. No correlation was present between serum DBH activity and systolic (r = -0.01, NS) or diastolic (r = +0.02, NS) blood pressure. No significant differences in serum DBH activity was observed between individuals with blood pressure in the lower, middle or upper deciles. Serum DBH activity was similar in subjects with normal blood pressure, in individuals with widely fluctuating blood pressure and in patients with fixed hypertension. The results suggest that serum DBH activity cannot be used as an aid in the diagnosis of essential hypertension of middle-aged men.


The importance of vasopressin in the development and maintenance of DOC-salt hypertension in the rat.

Experiments were performed to determine the role of vasopressin in deoxycorticosterone (DOC)-salt hypertension. In order to determine if vasopressin is necessary for the development of DOC-salt hypertension, rats with hereditary diabetes insipidus (DI) and normal Long-Evans rats (LE) were unilaterally nephrectomized, treated with DOC Pivalate (30 mg/kg . week) and given saline to drink for 8 weeks. A second group of DI rats were unilaterally nephrectomized, but received no treatment. Systolic blood pressure (SBP) increased 40 mm Hg in the LE group (p less than 0.01) but failed to increase significantly in either DI group. Urinary excretion of vasopressin (UADHV) and SBP were measured in unilaterally nephrectomized LE rats treated with DOC and salt (DOC-LE), salt alone (NaCl-LE) and untreated rats (H2O-LE). The UADHV was elevated in DOC-LE (p less than 0.01) and NaCl-LE (p less than 0.05), but only the DOC-LE rats became hypertensive. Finally, the I.V. injection of analogs of vasopressin, which block its pressor but not antidiuretic activity, lowered mean arterial blood pressure 27 +/- 5 mm Hg in 11 conscious DOC-salt hypertensive rats. It is concluded that vasopressin plays a major role as a pressor agent in both the onset and maintenance of DOC-salt hypertension.


Metastasis of human tumors in athymic nude mice.

The incidence of metastasis of xenogeneic tumors transplanted to nude mice is controversial. We studied 106 malignant human tumor lines in a total of 1,045 nude mice, and observed metastasis in only 14 instances (1.3%), involving 11 different tumor lines. Three of the lines showed repeated metastasis. Breast tumor lines metastasized with significantly greater frequency than other tumor types. None of the sarcoma lines metastasized. Tumors derived from human metastases were no more prone to metastasizing in nude mice than were tumors derived from primary sites. However, deep penetration of the body wall during growth of the tumor transplant was highly correlated with metastasis (p less than 0.001). Such factors as nude mouse health, tumor size and growth rate, and age and sex of the host mouse were not correlated with metastasis. Serial passage in nude mice did not select for a more malignant tumor line, since the incidence of metastasis did not differ at various passage levels. Thus, metastasis of human malignant tumors in nude mice would appear to depend primarily upon the site of tumor growth in the nude mouse, and upon the intrinsic metastasizing capability of the tumor line employed.


Content and distribution of macrophages and lymphocytes in solid malignant human tumours.

The relative content of lymphocytes and macrophages was studied in 20 solid malignant human tumours. Mechanically prepared tumour-cell suspensions contained both lymphocytes (0.2-4.0%) and macrophages (0.2-7.0%). Macrophages were characterized as esterase-positive, phagocytosing cells. The distribution of the cells within the tumours was studied on cryostat sections. A modified esterase method, using alpha-naphthyl butyrate as substrate, always gave an intensive staining of tissue-infiltrating macrophages, even in malignant tissues where some neoplastic cells exhibited a weak to moderate esterase activity. Tumour-infiltrating lymphocytes were studied in HE-stained tissue sections. The study demonstrates that both cell types were usually more numerous in the stroma surrounding the tumour tissue and in the stromal septa between the cords of malignant cells (peripheral infiltration) than in the central areas of the tumours. No signs of cell necrosis were seen near the lymphocyte zones or the stromal macrophages; however, some central necrotic areas contained aggregates of macrophages, lymphocytes and PMN.


Embryo-derived teratocarcinoma: I. The role of strain and gender in the control of teratocarcinogenesis.

The role of gender and genetic (strain-specific) factors in the regulation of teratocarcinogenesis was studied by monitoring the outgrowth of benign and malignant embryo-derived teratoid tumors, i.e., teratomas and teratocarconomas in several mouse strains. Teratocarcinomas were produced in all mouse strains tested, but the ratio of teratoma to teratocarcinoma varied from one strain to another. A high yield of teratocarcinomas was obtained in A/J, BALB/cJ, DBA/2J, CBA/J and C3H/J mice, irrespective of the sex of the recipient. A low yield of teratocarcinomas was obtained in both male and female C57BL/6J and AKR/J recipients, and in 129/J female recipients. For all strains but 129/J and eventually AKR/J the sex of the recipients did not significantly affect the outgrowth of embryo-derived tumors. These data suggest the existence of mouse strains with high and low permissiveness for embryo-derived teratocarcinogenesis. The sex of the recipients may influence the yield of embryo-derived teratocarcinomas in some mouse strains but is of no consequence in others.


Immunotherapy of an ascitic rat hepatoma with cord factor (trehalose-6, 6'-dimycolate) and synthetic analogues.

The ability of cord factor (trehalose-6, 6'-dimycolate) and a range of shorter carbon chain fatty acid trehalose diesters to suppress growth of an ascitic rat hepatoma has been examined and compared with that of whole, living BCG organisms. Aqueous suspensions of BCG, and cord factor in 0.4% arachis oil:Triton emulsion, injected intraperitoneally, retarded growth of up to 10(5) ascites tumour cells. Trehalose-6, 6'-dibehenate was also tumour-suppressive, but only against lower challenge inocula (10(4) cells). Trehalose-6, 6'-dipalmitate and 6,6'-di-0-2-tetradecyl -3-hydroxyoctadecanoyl alpha, alpha trehalose (designated C76) were virtually ineffective and 6,6' -di-0-2-eicosyl-3-hydroxy-tetracosanoyl alpha, alpha trehalose (designated C100) gave small and variable effects only against low challenge inocula. However, improved responses were seen with light mineral oil in place of arachis oil in the emulsions.


Lung carcinoma induction in BD rats after a single intratracheal instillation of an arsenic-containing pesticide mixture formerly used in vineyards.

A single intratracheal 0.1-ml instillation of a calcium-arsenate containing vineyard pesticide induced multi-focal bronchogenic adenocarcinomas and bronchiolar-alveolar-cell carcinomas in nine out of 15 treated BD rats. This seems to be the first experimental confirmation of carcinogenicity of arsenic-containing pesticides used formerly in vineyards.


Immunological basis for differences in susceptibility to hydrocarbon oncogenesis among mice of a single genotype.

The tendency for tumor induction to occur preferentially in certain individual mice of an inbred strain vanished when the putative immune response was reduced either by radiation plus thymectomy or by reduction of the concentration of the chemical oncogen.


Alterations in chromosomal proteins in C3H/HeN mice with spontaneous primary hepatocellular carcinomas.

The chromosomal proteins from chromatins of normal and background liver and spontaneous primary hepatocellular carcinomas (PHC) of C3H/HeN mice were examined by high resolution acid-urea and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. Before gel analysis, the histone and non-histone chromosomal proteins (NHCP) from the various tissues studied were separated by affinity chromatography. No qualitative or quantitative differences in histone content were detected in any of the chromatins studied. Tumor and background chromatins did, however, exhibit higher NHCP content than normal tissues. When NHCP were fractionated on SDS polyacrylamide gels, the appearance of 10 new unique NHCP, many of which were of high molecular weight, was found to accompany the manifestation of malignancy in these C3H/HeN mice. Conversely, the abundance of a number of individual NHCP decreased precipitiously in PHC chromatin; and, additionally, two high-molecular-weight NHCP present in normal chromatins were not detectable in background and PHC chromatin. A number of NHCP were found to be unique to each of the chromatins studied. The progressive increase in total number of NHCP in background and PHC chromatin when compared to normal chromatin suggested that such changes in NHCP might indicate progression towards malignancy. The acquisition of new high-molecular-weight NHCP, the loss of some high-molecular-weight NHCP, and the decrease in individual NHCP found for mouse PHC correlate well with findings of previous studies in other systems in which malignant transformation was induced with some agent. It appears then that changes in NHCP accompany both spontaneous and induced malignant transformation, and that such changes may reflect alterations in gene expression known to accompany malignancy.


Betel quid chewing and oral cancer: experimental studies on hamsters.

Betel quid ingredients--betel nut, betel leaf, lime, catechu and tobacco--were tested separately and in various combinations for carcinogenicity, using hamster cheek pouch as the experimental site. The four modes of administration used were (1) tri-weekly painting of the cheek pouch with aqueous extracts of test materials, (2) deposition of replaceable wax pellets containing the test material, (3) gelatin capsules containing the powdered material and (4) insertion of natural material into the pouch for trauma and direct exposure. Untreated controls and standard carcinogen DMBA-treated controls were also maintained. A total of 317 young adult golden Syrian hamsters (Mesocricetus auratus) used for the experiments were killed in two age groups: 6-12 months and 13-24 months, only when signs of general debility were observed. In the untreated controls, animals were free of any malignancy. In the experimental series, various betel quid ingredient combinations under test induced both oral and gastric lesions ranging from massive atypia and precancerous lesions to frank carcinomas. Maximum lesions were observed in the groups receiving betel nut, lime and tobacco combinations and in the polyphenol fraction of betel nut containing major tannins. The mode of administration of test material resulted in distinct differences; tri-weekly paintings giving oral lesions in the range of 22-23% and gastric lesions 39-48%; the same material given either through the replaceable gelatin capsule or in natural form induced 69% oral lesions and 63 to 82% gastric lesions. Overall evaluation of the data of all the four series confirms the potent carcinogenicity of betel nut, particularly its tannin-containing polyphenolic fraction and its combination with lime and tobacco. Maximum oral lesions induced in the hamsters by continuous exposure to capsules and natural material, highlight the direct relationship of frequency of chewing in habitual chewers with oral carcinogenesis. The high incidence of gastric (forestomach) lesions invites special attention.


Estimation of catecholamines in human plasma by ion-exchange chromatography coupled with fluorimetry.

Estimation of catecholamines in human plasma was made by ion-exchange chromatography coupled with fluorimetry. Catecholamines in deproteinized plasma were adsorbed onto Amberlite CG-50 (pH 6.5, buffered with 0.4 M phosphate buffer) and selectively eluted by 0.66 M boric acid. The catecholamine fraction was separated further on a column of Amberlite IRC-50 which was coupled with a device for the automated performance of the trihydroxyindole method (epinephrine and norepinephrine) or the 4-aminobenzoic acid-oxidation method (dopamine). One sample could be analysed within 25 min with either method. The lower detection limits were 0.02 ng for epinephrine and dopamine, and 0.04 ng for norepinephrine. Plasma catecholamine contents of healthy adults at rest were epinephrine 0.07 +/- 0.01 ng/ml (n = 19), norepinephrine 0.27 +/- 0.03 ng/ml (n = 19) and dopamine 0.22 +/- 0.03 ng/ml (n = 26). The procedure of adsorption and elution of the plasma catecholamines by ion-exchange resin was simple, the simplicity contributing to constant recovery. The catecholamine fraction could be analysed without evaporation of the eluate. The analytical column could be used for the analysis of more than 1000 samples before excessive back-pressure developed. Our method of continuous measurement of plasma catecholamine fulfils clinical requirements.


Selected ion monitoring assay for biogenic amine metabolites and probenecid in human lumbar cerebrospinal fluid.

Details are presented of an improved selected ion monitoring assay for the major biogenic amine metabolites and probenecid in human lumbar cerebrospinal fluid (CSF). The metabolites and probenecid are simultaneously extracted with ethyl acetate from an acidified aqueous phase, and are simultaneously converted to pentafluoropropionyl esters by reaction with pentafluoropropionic anhydride and pentafluoropropanol. The esters of the metabolites are analyzed following a single injection of the derivatized sample onto the gas chromatographic column, while the ester of probenecid is analyzed following a separate injection onto the gas chromatographic column. Quantitation is achieved using for internal standards dueterated analogues of the metabolites and a chemical analogue of probenecid. Data are presented on the concentration of free and conjugated forms of the metabolites in lumbar CSF taken from healthy volunteers.


Rapid assay for theophylline in clinical samples by reversed-phase high-performance liquid chromatography.

A fast, sensitive and highly specific method for the determination of theophylline in human serum is reported. Using a C18-bonded reversed-phase column with an acetonitrile-acetate buffer mobile phase theophylline is completely resolved not only from other dietary xanthines and their metabolites but also from co-administered drugs such as paracetamol and phenobarbitone. Use of beta-hydroxyethyltheophylline as internal standard allows a within batch precision of 2.0% and a between batch variation of 3.0%. Factors involved in the development of the method and its performance are discussed.


Evaluation of a modified high-performance liquid chromatography assay for acebutolol and its major metabolite.

Extensive modification of an existing high-performance liquid chromatography assay for acebutolol and its major metabolite has markedly improved chromatographic stability eliminating the previous need for frequent adjustment of the eluent composition to accommodate continuous loss of column retention. The eluents now used and avoidance of the requirement for elevated column temperature may be significant factors in the ability to maintain column life over 8 months of continuous use with little decrease in retention. As a result of the improved chromatographic stability full advantage can now be taken of automatic injection devices for the unattended processing of large numbers of samples. A significant modification of the work-up of blood samples has improved precision of the assay in whole blood. Nevertheless, it is recommended that plasma samples rather than whole blood be analyzed, since the plasma assay is faster and still more precise.


Trace analysis of the MIF analogue pareptide in blood plasma by high-performance liquid chromatography and short-wavelength excitation fluorometry.

A high-performance liquid chromatographic procedure was developed and applied to analysis of the pharmacologically active MIF analogue pareptide in human plasma. The procedure involves formation of a fluorescent 7-chloro-4-nitrobenzyl-2-oxa-1,3-diazole (NBD-Cl) pareptide derivative followed by separation of the NBD derivative from plasma components on a 30-cm microparticle octadecylsilane bonded column. The separated derivative was quantitated using a short-wavelength excitation fluorometric detector. The detection limit of pareptide in plasma samples was 5 ng or 17 pmoles per ml of plasma. In the absence of plasma, the corresponding on-column detection limit was 0.5 pmoles.


[Separation and characterization of acidic urine constituents (author's transl)].

The acidic compounds of urine were separated by thin-layer chromatography in eight fractions. Each fraction was investigated separately by the combination glass capillary gas chromatography-mass spectrometry. About 500 compounds were detected, 2/5 of these could be characterized by their mass spectra. Retention data and key fragments of the mass spectra were tabulated. Many of the detected compounds are still unknown.


[Occurrence of alpha-alkyl-substituted malic acids, and beta-hydroxy-beta-alkyl-substituted dicarboxylic and tricarboxylic acid derivatives in normal urine (author's transl)].

Urine contains a number of alpha-hydroxy acids so far unknown to occur in biological liquids. Besides the already as urine constituent known methylmalic acid, also the ethyl, isopropyl and butyl derivatives of malic acid were found. Further metabolites in urine are a beta-propyl-substituted beta-hydrosyglutaric acid, a beta-hydroxy-beta-[methyl-carbomethoxy]-adipinic acid and two isomeric alpha-methylcitric acids.


Automated quantitative gas-liquid chromatography of intact lipis. II. Accuracy, precision and reproducibility of results.

The effect of various factors on the precision and accuracy of the gas chromatographic determination of neutral lipids was studied in the concentration range where the correction factors are dependent on the amount analyzed. The mutual effect of individual components of the neutral lipid spectrum on the recovery was examined. A method is described which provides the stable recovery of the components present at low concentrations, using the addition of high-molecular-weight triglyceride (triarachidin) which does not interfere in the determination of the usual triglycerides. The validity of the correction factors measured with pure compounds was verified by hydrogenation of biological samples of various compositions. Hydrogenation of the sample also solves the problem of the determination of the triglyceride fraction of carbon number 46, which interferes under normal conditions with the determination of the cholesteryl ester fraction of carbon number 47. A method for the standardization of the gas chromatographic determination of neutral lipids is given using pure compounds instead of lyophilized biological samples. Long-term quality control was carried out using synthetic control samples. The results show sufficiently low values of the variation coefficients over the whole period. The values of the variation coefficients measured over an interval of 25 weeks are about 4% for the main components of the neutral lipid spectrum and 6.3% for the components present at concentrations up to 5%. Thw within-day variation for the most neutral lipid fractions and for lipid classes attains a value of 40-75% of the day-to-day variation. The most satisfactory values are obtained for the variation within a single series which amount to less than 2% for all substances except for triglyceride fractions 48 and 54. The correlation of the determination of total cholesterol and triglycerides by gas chromatography and by enzymatic methods shows a very good agreement between the results obtained by the two methods. Using quality control, it is possible to follow the accuracy of the calibration and to demonstrate objectively the necessity for system recalibration.


Determination of phenylalanine in serum using reversed-phase liquid chromatography and fluorescence detection.

A liquid chromatography procedure is reported for determining phenylalanine in small volumes of serum. A 10-microliter volume of serum was deproteinized with ethanol and an aliquot was derivatized with dansyl chloride reagent. The dansylated phenylalanine and the norleucine internal standard were separated using reversed-phase chromatography and measured with a fluorescence detector. Linearity was excellent over the range 50-800 mg/l. Within-run precision was better than 4%. Total analysis time including chromatography was approximately 40 min. As little as 300 pg of dansylated phenylalanine was detected.


Amniotic fluid uric acid levels determined by reversed-phase liquid chromatography with spectrophotometric and electrochemical detection.

A rapid and precise, reversed-phase high-performance liquid chromatography method for amniotic fluid uric acid is described. Detection of uric acid and other naturally occurring constitutents is based on UV absorption at a wavelength of 280 nm and direct electrochemical oxidation at a potential of +0.800 V. The total analysis time is short (20 min) and the assay requires only filtration of the samples. Uric acid levels were determined in 14 samples of amniotic fluid obtained during the 15th to 24th week of gestation. Results ranged from 0.897 to 4.39 mg per 100 ml of amniotic fluid.


Determination of amoxicillin in body fluids by reversed-phase liquid chromatography coupled with a post-column derivatization procedure.

Quantitative methods for determination of amoxicillin in body fluids are described. They comprise separation by reversed-phase chromatography (LiChrosorb RP-8, 5 micron) of the aqueous supernatants obtained from plasma or urine after purification steps involving protein precipitation followed by extraction in the case of plasma, or a double extraction procedure in the case of urine, post-column derivatization with air segmentation, and finally measurement of the UV absorbance at 310 nm. The derivatization involves formation of the mercuric mercaptide of penicillenic acid and is specific for compounds with an intact penicillanic acid ring system. Detection limits achieved on injecting 200 microliter of plasma and 20 microliter of urine are about 25 ng/ml and 200 ng/ml, respectively, but it is possible to improve the sensitivity further by injecting larger volumes. Precisions (srel) obtained for determination of 0.10 and 0.45 migrogram/ml in plasma were 3.72 and 1.40%, respectively. Some problems regarding column stability originating from the injection of biological samples are discussed.


Sensitive electron-capture gas-liquid chromatographic assay for the de-ethylated metabolite of metoclopramide.

A procedure is described for determining 4-amino-5-chloro-2-methoxy-N-(2-ethylaminoethyl)benzamide, a metabolite formed by de-ethylation of metochlopramide, in urine from rats. The sampe is extracted (at pH approximately 13) with chloroform and, after treatment with heptafluorobutyric anhydride, derivatives of the extracted compounds are analyzed by gas-liquid chromatography, with electron-capture detection and diazepam as internal standard. The drug and its metabolite are separated, and the latter can be determined in the range 0.4 to 1.85 microgram/ml in the sample. The behaviour of the metabolic during chemical-ionization and electron-impact mass spectrometry is discussed.


Determination of L-alanosine in plasma and urine by reversed-phase high-performance liquid chromatography of the Dns derivative.

L-Alanosine is an antitumour antibiotic that has recently been placed in clinical trial. We have developed a relatively rapid and specific assay for urinary and plasma alanosine, based on formation of the Dns derivative and separation of this from other Dns compounds by reversed-phase high-performance liquid chromatography. Dns-Alanosine is detected by its absorption at 254 nm, since alanosine is atypical in that it forms a Dns derivative with very low fluorescence. The lower limit of detection of alanosine in plasma is 0.1 microgram/ml. The assay has been used to measure the levels of alanosine in the plasma and urine of rabbits and of man.


Separation of nebramycin components by thin-layer chromatography.

A thin-layer chromatographic method has been developed for the detection of major nebramycin components, for the separation of tobramycin from other components and for studying the hydrolysis of carbamoyl derivatives and procedures for isolation and purification. A sensitive method has also been established for the detection of kanamycin B in tobramycin and for the assay of apramycin in kanamycin B.


Gas-liquid chromatography of digitoxigenin and digoxigenin as acetates of their epoxygeninic acid methyl esters.

Gas chromatographic analysis of ditigoxigenin and digoxigenin, the genins of the cardenolide glycosides digitoxin and digoxin, cannot be done without derivatization. However, during the derivatization, side-reactions often present serious problems. A procedure has been found for transforming digitoxigenin and digoxigenin into the corresponding acetates of their epoxygeninic acid methyl esters, which are stable compounds and suitable for gas chromatographic analysis.


Characteristics of lithium iodide-containing poly(ethylene glycol) as a gas chromatographic stationary phase, and its application to analysis of amidic drugs.

The characteristics of lithium iodide-containing poly(ethylene glycol) as a gas chromatographic stationary phase have been evaluated in terms of partial free energy of transfer (delta G t0) from poly(ethylene glycol) to the lithium iodide-poly(ethylene glycol) system for a variaty of amides (n-fatty acid amides, lactams, benzamides, anilides, nicotinamides, isonicotinamides, barbiturates, pyrazolones) and several amines. The changes in relative retention and resolution of two solute peaks caused by the addition of lithium iodide to poly)ethylene glycol) are correlated with the difference in their delta Gt0 values. The application to the specific separation of some amidic drugs is demonstrated.


Bacteriophage typing scheme for Salmonella infantis.

A bacteriophage typing system is described for Salmonella infantis. Nine phages were selected, of which three were isolated from sewage and six from human feces. All except 7 of the 546 strains collected between 1974 and 1978 could be classified into 23 different phage types. The five most common phage types comprised 26, 13, 9, 9, and 9% of all strains, respectively. Strains from humans, animals, food, and water isolated during nine episodes, or from given patients at different intervals of time, belonged to the same phage type.


Evaluation of commercial latex agglutination reagents for grouping streptococci.

A total of 155 strains of beta-hemolytic streptococci were serologically grouped by conventional techniques (Lancefield extraction and capillary precipitin testing) and by latex agglutination (LA). Agreement between conventional and LA techniques was 97% when the instructions of the manufacturer for the LA technique were followed. Agreement of 99% was obtained when modified autoclave extracts were used as antigens in the LA procedure. A total of 82 strains of non-beta-hemolytic streptococci were also tested by conventional, prescribed LA, and modified autoclave procedures. The agreement between conventional techniques and both LA procedures was 76%. However, when serological cross-reactions in the conventional grouping procedures were considered as errors, the accuracy of identification of both LA procedures was 88% among the non-beta-hemolytic strains. Of 13 strains of Streptococcus bovis, 10 did not react with the LA group D reagent but were serogroup D by conventional techniques. More S. bovis strains were grouped by the LA technique when extracts of 20 ml of broth cultures were used as antigens; however, cross-reactions were observed with non-group D strains when this technique was applied to them.


Destruction of bacterial viruses in serum by heat and radiation under conditions that sustain the ability of serum to support growth of cells in suspended culture.

Methods for inactivating bacterial viruses in serum were developed through the use of heat and ionizing radiation, and the effects of these treatments on the growth rates of cultured cells were tested. Viruses chosen for this study were the radiation-resistant bacteriphage f2 and heat-resistant phage T4. The viabilities of these phages were reduced more than 2 and 4 orders of magnitude, respectively, by a treatment at 60 degrees C for 30 min followed by 420 krads of ionizing radiation. Simultaneous application of heat and radiation caused a considerably greater reduction in viability of both phages in serum, but also caused a significant decrease in the growth rates of L cells in medium supplemented with serum treated in this manner. Treatment of serum with these same doses but given in the sequential fashion of heat followed by radiation caused little or no change in the growth rates of L cells. Finally, it was found that simultaneous treatment of serum with these doses of heat and radiation had little effect on the growth rates of either HeLa or Chinese hamster cells.


Evaluation of vented blood culture media with sorbitol.

Two consecutive studies comparing the recovery of microorganisms from transiently vented blood cultures in tryptic soy or brain heart infusion broth with added sorbitol and that from tryptic soy broth without sorbitol failed to demonstrate any significant advantage of hypertonic media. A significantly greater number of organisms was isolated in both studies from the medium without sorbitol.


Relative recovery of anaerobes on different isolation media.

The recovery of clinical anaerobic isolates on selective and nonselective agar media, as well as the time required to detect the isolates, was examined. Of a total of 235 isolates, 77, 46, and 40% were detected on Schaedler blood agar, colistinnalidixic acid blood agar, and kanamycin-vancomycin-lysed blood agar, respectively, and 94% were detected on the combination of Schaedler blood agar with kanamycin-vancomycin-lysed blood agar. A total of 19% of the anaerobes were detected after incubation for 1 day, and 70% were detected after 2 days.


Upper limit of normal titer for detection of antibodies to Legionella pneumophila by the microagglutination test.

An upper limit of normal microagglutination titer for the Knoxville strain of Legionella pneumophila was determined by testing 830 sera. The upper limit of normal titer was found to be 8; however, we suggest that when applied to single serum specimens a titer of 32 be considered the lowest titer suggestive of infection (not necessarily disease).


Group G streptococcal pneumonia and sepsis in a newborn infant.

A case of neonatal pneumonia and sepsis caused by a group G Streptococcus is described. Clinical and microbiological aspects of group G streptococci are compared with those of group B streptococci.


[Lactic acid in the human umbilical cord. Influence of glycolysis in the blood in cord and in cordonal tissue (author's transl)].

The blood staid in human umbilical cord increases his lactate concentration (about + 4 mg/l/mn, in average). It is the fact of an high metabolic activity of foetal erythrocytes for glycolysis, but also of action of cordonal tissue itself. This tissue, after elimination of blood, allows an high concentration of lactate, which increases when cord is kept at room temperature, reaching and rising above 1 or 2 g/kg, after many hours. This glycolytic process indicates than cord tissue has a metabolic activity which seems to be in relation with the high lactate concentration of the amniotic fluid.


[The correlation between the results of caryotype studies and sperm counts. 119 cases (author's transl)].

119 patients had sperm counts and caryotyping performed. Seven out of 33 azoospermic patients had a major caryotype abnormality of the gonosomes. Five out of 86 subjects who were not azoospermic had a persistent translocation and other moderate abnormalities in the sperm picture.


[Antenatal diagnosis of the sex of the fetus by diagnostic ultrasound (echography) (author's transl)].

The authors present the results they obtained in diagnosing fetal sex in utero by the use of real-time echography. They also have reviewed the other different techniques for the diagnosis and their fallibility. The prediction of fetal sex is possible in 90 p. 100 of echographies carried out routinely. When a very careful well codified technique is used the method becomes highly reliable. In a successive series of 103 pregnancies fetal sex was diagnosed correctly in every case. The authors point out that they diagnose female sex through positive echographic finding and not by a process of exclusion of the male organs.


[Oxytocinase: levels in normal and pathological pregnancies].

The author has determined the levels of plasma oxytocinase in a series of normal and pathological pregnancies. It appears from studying these levels that it is useful to measure the oxytocinase particularly in cases where there is failure of intra-uterine growth.


[Necrotising enterocolitis and pregnancy. One case (author's transl)].

Having had a case of necrotising enterocolitis which occured on the third day after delivery and which was successfully treated by total colectomy with secondary restoration of bowel continuity, the authors discuss the pathogenicity of this condition. It can be secondary to a haemorragic or a septic obstetrical complication. It may equally be the sequel of a functional paralytic ileus which, although rare, is one of the classical complications following labour and Caesarean operations. In this way it can be compared to those cases of enterocolitis which are secondary to ileus, such as are found in patients who have a tumour of the colon or in patients who have been treated with neuroleptic drugs.


[Tumours of the ovary in young women. The way they arise and the limits of conservative treatment (author's transl)].

Although tumours of the ovary are rare in woman under the age of 35 they are not exceptional and they form 15 p. 100 of the cases (17 patients) out of a series of 112 tumours of the ovary which were collected in 7 years. 11 of these patients had only one ovary affected and were able to be treated conservatively. In this way 5 of them were able to carry one or more pregnancies to term and another patient who was lost from observation for two years had to have a second operation to deal with a recurrence in the opposite ovary. Conservative treatment is only permissible if one is absolutely sure that the contro-lateral ovary is normal, and if there is the slightest doubt ovariotomy should be carried out to perform a biopsy. Furthermore the authors suggest that in view of the difficulties of strict supervision for a long term the opposite ovary and tube should be removed and hysterectomy carried out once the patient has had one or two children or when they have reached the age of 40.


[A summary of the results of microsurgical exploration and treatment. 40 cases of male sterility (author's transl)].

The authors report their statistical results of 40 subjects that were explored and where double epididymo-deferens anastomoses were achieved. They found that there were associated conditions which worsened the prognosis, namely: stenosis of the genital tracts and broncho-pulmonary infections. On the other hand stenosis of the genital tracts associated with varicocoele gave a better prognosis. Permeability was achieved in 40 p. 100 of cases. Six pregnancies occured from 33 patients who were followed up with post-operative spermograms. The following treatments are useful if added to the surgical treatment: gonadotrophins, corticoids and antibiotics. They point out that surgical cure is much more likely when varicocoele is associated.


[Study of transplacental transmission of beta-lactam antibiotics (author's transl)].

The authors present the results of the study of transplacental transfer of 3 recent beta lactam antibiotics: amoxicillin, cephradin and cefoxitin. The experiment was carried out in 37 pregnant women who had a premature rupture of membranes or who were undergoing a cesarean section. In these patients antibiotherapy was indicated in view of a possible superinfection. The results of the study show a worthwhile transplacental penetration of the 3 drugs tested. The concentrations achieved in umbilical cord blood and amniotic fluid increase with time and finally exceed the levels measured in maternal blood, after a period of time varying from 2 to 4 hours according to the drug and the route of administration.


Group graph of the genetic code.

The genetic code doublets can be divided into two octets of completely degenerate and ambiguous coding dinucleotides. These two octets have the algebraic property of lying on continuously connected planes on the group graph (a tesseract) of the Cartesian product of two Klein 4-groups of nucleotide exchange operators. The K X K group can also be broken into four cosets, one of which has completely degenerate coding elements, and another that has completely ambiguous coding elements. The two octets of coding doublets have the further algebraic property that the product of their internal exchange operators naturally divide into two exactly equivalent sets. These properties of the genetic code are relevant to unraveling error-detecting and error-correcting (proof-reading) aspects of the genetic code and may be helpful in understanding the context-sensitive grammar of genetic language.


Hereditary macrostomus in the rabbit: a model for Treacher Collins syndrome, one form of mandibulofacial dysostosis.

Macrostomus in the rabbit was diagnosed externally by the presence of papillae at the corners of the mouth. Internally the effect is on the zygomatic complex and is extremely variable. Inheritance appears to be due to a single autosomal recessive gene with incomplete penetrance. We propose the symbol mst for the gene responsible for this condition. Evidence is presented to suggest that this gene is very likely in linkage group IV. Macrostomus in the rabbit is a model for the Treacher Collins syndrome, one form of mandibulofacial dysostosis in humans.


[A study of the obstacles to reduction in congenital dislocation of the hip (author's transl)].

Investigation on ten patients with typical congenital dislocation of the hip generalized joint laxity indicated the changes in the antero-inferior part of the acetabulum, which are mainly adhesion and narrowing produced by the capsule pulled superoposteriorly, as the major obstacle for successful reduction. Inverted limbus, pulvinar mass and ligamentum teres may represent secondary obstacles mostly due to iatrogenic procedures. The purpose of the closed reduction is to get rid of the obstacles during the conservative treatment. The cause of the failure of conservative treatment of congenital dislocation of the hip is a misjudgement of true reduction, allowing enough cavity for the femoral head in the antero-inferior part of the joint without the presence of the capsule between the femoral head and the acetabular cartilage. To get better understanding of the true reduction, tangential view in the arthrography of the hip joint gives us more precise data about the antero-inferior part of the joint because the acetabulum faces the joint cavity laterally and anteriorly.


[Radiographical and anatomical studies on the pathogenesis of degenerative spondylolisthesis (author's transl)].

In order to elucidate the causes of degenerative spondylolisthesis (D.S), the functional role of lumbar intervertebral joints and also laminae of the vertebral arch was studied by means of roentgenographic and also pathological exams. The inclination of the laminae of the vertebral arch and the intervertebral joints changed with age in accordance with statokinetic loading of the lumbar spine. In the group with increased lumbar lordosis at the upper level, movement of the 3rd and 4th lumbar vertebrae was great, accompanied by horizontal inclination of the laminae of the vertebral arch and the intervertebral joints and by osteoarthritic changes (O.A). This is similar to D.S. D.S is caused by abnormal spinal curvature and structural specificity of the lower limbs vertebrae, resulting in degeneration of the laminae of the vertebral arch and the intervertebral joints, changes in the inclination, decreased control of movement, and sliding of the vertebrae. Prodromal conditions resulting in the sliding of the vertebrae and in disc degeneration appear to be caused by D.S.


[Classification and growth of loose bodies in joints (author's transl)].

A loose body in a joint can be attributed to various factors, such as osteochondritis dissecans, osteochondral fracture, synovial chondromatosis, or fracture of osteophytes in osteoarthritis. Some of the cases involving loose bodies in joints have evident etiological factors identified on the basis of radiographic and/or operative findings, while the others have etiological factors which have not yet been clarified. We have therefore investigated loose bodies taken from joints at arthrotomy and attempted to establish the process involved in their growth by histopathological, microradiographic and fluorescence-microscopic, and scanning electron microscopic means, and tried to clarify whether it is feasible to assess the etiological factors from these findings. The results revealed that the loose body in the joint is surrounded eventually by fibrous tissue, the lower layer of which forms osteo-cartilaginous tissue and that, in the center of the loose body, degenerated osteocartilaginous tissues from which etiological factors can be suspected are located, and the author mentioned that etiological diseases of the loose body could possibly be suggested from the studies on their microfine structures. It was furthermore clarified that the loose body is fed by synovial fluid and grows, and yet the rate of the growth is extremely slow.


[An experimental study on the effects of the plating method for bone fractures--with reference to bone union and bone strength (author's transl)].

The plating method for bone fractures is in wide use. The author studied both new bone formation and bone atrophy of the cortex of fracture site after long periods of plating. This study is concerned with the bone strength of the fracture site as influenced by rigid plate fixation, and also with bone union after the removal of the plate. The experimental results, using mature rabbits as subjects, showed the following: X-rays of the bone union at the fracture site treated by the plating method confirmed evident union at 8 weeks after surgery. The bone strength at the region of this union, as tested by the bending method (an Instron Type Testing Machine was used), showed its highest values at 28 weeks after surgery. Yet, this was only 60% of the bone strength as measured at a non-fracture site; it then gradually declined in strength. On the other hand, in the case where the plate had been removed 8 weeks after surgery, bone strength was recovered to the same level as that of non-fracture sites 16 weeks after surgery. In conclusion, rigid plate fixation for a long period may result not only in progressive bone atrophy of the cortex but also in a delayed remodeling of the bone at the region of the bone union. It is, therefore, very important to remove the plate as early as possible once union has been achieved. Careful observation, however, is necessary during the short time between plate removal and the period when bone strength becomes again normal.


[An experimental study on the influences of artificial socket upon articular cartilage of the femoral head (author's transl)].

In order to investigate the influences of artificial socket made of stainless-steel, pure titanium, high-density polyethylene (HDP), and polycrystal ceramics upon the articular cartilage of the femoral head, experimental artificial sockets were developed. This artificial socket was inserted into the hip joint in a dog, and the changes in the articular cartilage of the femoral head were studied at varying intervals after insertion of the socket. The results revealed that both the stainless-steel and the ceramic sockets caused slight atrophy and degeneration in the articular cartilage of the femoral head since one month after the operation, and these changes were increasingly more appreciable as time advanced. Any changes were hardly noted in the articular cartilage of the femoral head by titanium socket at the end of one month after the operation, whereas the HDP socket caused substantial changes. At the end of one year after the operation, marked degeneration was noted in the articular cartilage of the femoral head by socket made of stainless-steel, titanium, and HDP, and also the femoral head developed deformity. On the basis of the scanning electron microscopic findings on the surface of the articular cartilage of the femoral head affected by each of these artificial socket, and of the metal microscopic examinations of the surface of the artificial sockets, and furthermore on the basis of the findings of gradual deformation of femoral heads due to the artificial sockets, it was shown that articular cartilage of the femoral head was mainly influenced in the case of metallic socket and that both sides of the artificial socket and the articular cartilage of the femoral head were worn out in the case of HDP socket. The ceramic socket was found cracked, developing a loosening in some cases. In summary, it was concluded that the articular cartilage of the femoral head is affected by the artificial socket at a relatively early stage after its application despite materials of the sockets.


[Problems concerning bone union influenced by heat generated in osteotomy (author's transl)].

The author experimentally investigated in rabbits how the heat generated by bone saws affected bone union in osteotomy. Radiographic and soft X-ray findings revealed that recovery of the lesion in every bone shaft after osteotomy was marked in all. However, the time required to obtain complete recovery of the lesion differed depending on the instruments used. That is to say, it required 3 weeks after surgery by means of Gigli's saw, 4 weeks by means of Stryker bone saw and lower speed bur (2,000 rpm) and 6 weeks by means of higher speed bur (18,000 rpm). Recovery of lesion by bone cut with each bone saw on the unscalded side was one or two weeks earlier than that in the scalded side. This was also proved on histopathological findings. With regard to the disappearance of osteocytes in the bone cortex at the cut surfaces of bones, micrometric findings revealed that there was a great difference between the scalded and the unscalded side in every group except in the group where a Gigli's saw was used. However, almost no difference was observed on the unscalded side among every bone saw. According to scanning electron microscopic findings at every surface cut with various bone saws, collagen fiber was preserved on the unscalded side where physiological saline solution was sprayed. Particularly, the collagen fiber appeared to be torn off in the case of lower speed bur (2,000 rpm), the cut surface was well uniform in the case of higher speed bur (18,000 rpm). Thus, I have learnt that we do not need to give much attention to the kind of instrument, if effort using physiological saline solution, are made to avoid the heat generated by bone saws when osteotomy is done.


[A study on postoperative management and pre-and postoperative changes of blood volume in orthopedic surgery (author's transl)].

In order to apply adequate postoperative management of patients in the orthopedic surgery, the in vivo effects of the operations were investigated in the present study, from various viewpoints such as circulatory functions, metabolism of proteins, and hepatic and renal functions. In addition, the circulating blood volumes were determined in order to know the variations in the amount of blood loss from the operated regions during and after operations so as to improve the operative and postoperative management. On the basis of the results, the countermeasures against the adverse variations in blood volume were also investigated. Pediatric patients were found to be prone to develop more appreciable systemic postoperative reactions than those in adult ages, especially so in those of circulatory system, peripheral blood, hepatic functions, and protein metabolism, and also prone to be more affected by operative procedures. Senile patients were found to be prone to have less reserve capacity in circulation, peripheral blood, renal functions, and protein metabolism, and also prone to show delayed postoperative reactions and slower recoveries. The results of measurement of circulating blood volume in pre- and postoperations revealed that the loss of circulating blood in most patients was greater than the bleeding weight during the operation measured by gause count, and that, on the day following the operation, the blood volume was reduced by about 10-15% than those immediately after the operation. It was concluded that, as a whole, no changes causing serious deviation from the normal physiological ranges occur except in pediatric and senile patients. On the basis of the above findings, most appreciable systemic postoperative reactions of the operative procedures in orthopedic surgery would be attributed to by loss of blood volume during operations and that the postoperative hemorrhage from bone structure (which is usually difficult to control) or the postoperative hemorrhage after removal of air tourniquet would be almost negligible.


Spinal evoked potential (SEP) obtained by stimulation on the median nerve--experimental and clinical studies.

The present study was done experimentally and clinically to measure the segmental spinal evoked potential (SEP) of the cervical cord, stimulated on the median nerve, to try to determine the location and severity of cord lesion in patients with cervical spondylotic myelopathy. SEP in control subjects consisted of two waves. The primary reactive wave (R wave) consists of both the nerve root potential and the funiculus posterior potential. The secondary reactive wave (N wave) consists of the post-synaptic potential in the spinal cord. With regard to cervical spondylotic myelopathy, in the early stages the N-wave weakens or disappears at the level of lesion, then, as damage progresses, the R-wave also weakens or disappears.


Ultrastructural studies of certain aspects of the development of Trypanosoma congolense in Glossina morsitans morsitans.

The course of Trypanosoma congolense infections in Glossina morsitans morsitans was followed by electron-microscopic examination of ultrathin sections of the guts and proboscises of infected flies. Guts dissected from flies 7 days after infection with culture procyclic forms of T. congolense had heavy trypanosome infections in the midgut involving both the endo- and ectoperitrophic spaces. Trypanosomes were also seen in the process of penetrating the fully formed peritrophic membrane in the central region of the midgut. By post infection day 21, trypanosomes had reached the proboscis of the fly and were found as clumps of epimastigote forms attached to the  labrum by hemidesmosomes between their flagella and the chitinous lining of the food canal. Desmosome connections were observed between the flagella of adjacent epimastigotes. Flies examined at postinfection days 28 and 42 had, in addition to the attached forms in the labrum, free forms in the hypopharynx.


Starch gel electrophoresis: an effective method for separation of pathogenic and nonpathogenic Naegleria strains.

Isoenzyme electrophoresis of 7 different enzyme systems was used to compare 24 strains of Naegleria fowleri and 6 strains of N. gruberi. The 30 strains could be grouped into 4 distinct categories based upon zymogram patterns. No interstrain band variation in all enzyme systems was demonstrated in pathogenic strains of N. fowleri. Three nonpathogenic high temperature-tolerant strains of Naegleria had similar zymograms. Four of the 5 remaining nonpathogenic Naegleria strains had no interstrain band variation. Based upon zymograms, the 22 pathogenic strains constitute a homogenous species. Similarly the high temperature-tolerant nonpathogenic strains formed a cohesive group. The remaining nonpathogenic strains could be separated into 2 groups.


Purification of Plasmodium lophurae exoerythrocytic merozoites by an ion exchange column.

Exoerythrocytic merozoites of Plasmodium lophurae grown in embryonic turkey brain cells were successfully separated from host cell material by elution from a DEAE-cellulose column at ionic strength 0.22. Purity of parasite samples was assessed by sodium dodecyl sulphate acrylamide gel electrophoresis and electron microscopy. Increasing the ionic strength gave greater recoveries of merozoites, but host cell contamination increased.


Curative effects of the antipiroplasms amicarbalide and imidocarb on Trypanosoma brucei infection in mice.

The babesicides imidocarb and amicarbalide, which have structural similarities to the antitrypanosomatid diamidines, proved active against Trypanosoma brucei mouse infections: both cured infections when doses were administered daily for 3 days 24 h post-inoculation (curative dose imidocarb, 10 mg/kg; amicarbalide, 25 mg/kg). Mice were considered cured after survival 30 days longer than untreated infected controls, with no trypanosomes present in blood or cerebrospinal fluid smears. Both agents also cured when administered 48 and 72 h after challenge with T. brucei and prolonged the lives of animals 94 h after challenge. The results are discussed in respect to the potential of these carbanilides and their precursors, the antitumor phthalanilides, as lead compounds in chemotherapy of mammalian trypanosomiases.


An experiment in training in child care for general practitioners.

The educational objectives for training general practitioners are widely agreed. However, there is a shortage of paediatric posts in hospital, especially in Scotland. As there are obvious advantages in training clinicians who will work in general practice in the setting of general practice, we have initiated 10 posts in training each of six months' duration and each in addition to the trainee year. Three months will be spent in general practice itself and systematic teaching will also be provided for three months by the Edinburgh School of Community Paediatrics. Pre-course and post-course assessments will be carried out and the learning achieved will be evaluated.


The identification of children with learning problems in general practice.

Twenty-four childen who were patients in a large general practice in Southern England were seen in a learning problem clinic in the practice during a 14-month period. The method of assessment used took an hour per child but several examples show that it was both practical and effective.


Respiratory morbidity and exercise-induced bronchoconstriction in children.

A sample of 99 children had lung function tests before and after exercise. Sixteen had a decrease in lung function after exercise (forced expiratory flow) of more than 20 per cent, which indicated an important degree of exerciseinduced bronchospasm. Review of the medical records for the total sample revealed that those with exercise-induced bronchospasm had a higher consultation rate for respiratory symptoms, while their consultation rate for non-respiratory symptoms was the same as the remainder of the sample. These findings suggest that bronchial lability may be a predisposing factor in some respiratory diseases.


Food intake before migraine attacks in children.

A questionnaire survey of 120 children with migraine showed an average age of onset of 5.15 years, an equal sex ratio under nine years, and a positive family history in 79 per cent. Eye symptoms (42 per cent) and headaches (32 per cent) heralded an attack, with abdominal pain and vomiting later and less frequent. Visual aura was not recognized under five years, but occurred in 52 per cent of the 13 to 15 year age group. Most attacks occurred on schooldays and 82 per cent were over within two days.The 24-hour food intake before an attack was compared with the food intake seven days later when no migraine occurred. This suggested that fasting (41 per cent) or specific foods (38 per cent) could have been responsible for many of the attacks.


The tissue distribution and the pattern of excretion of [14C]-13-labeled 12, 13-epoxytrichothec-9-ene in mice and rats.

The distribution in the mouse tissues of 13-[14C]-12,13-epoxtrichothec-9-ene administered intravenously was determined by whole-body autoradiography and by tracing the radioactivity of the tissues oxidized in an Auto Sample Oxidizer. The appearance of the label in urine and feces was also followed by the tracer technique. The distributions of radioactivity in tissues as determined by the two methods were almost identical. On the autoradiograms of mice killed 10 min after the injection, marked blackening of the film was observed at the sites corresponding to the liver, kidney, and bladder with urine, and much less darkening at other sites. The radioactivities contained in the liver, kidney, urine and small intestine were 13.3, 2.3, 2.6 and 10.2% of the dose, respectively. The labeled toxin was rapidly excreted into urine and feces, 56.0 and 4.9% in 6 hr and 66.7 and 28.0% in 24 hr after injection, respectively. Oral administration of the labeled toxin to mother mice resulted in the appearance of radioactivity in the stomach contents of 7-day suckling mice, thus demonstrating indirectly the secretion of the toxin into the milk. An attempt to show a respiratory route of excretion in rats given the radioactive compound orally or intravenously failed to detect any radioactivity in the expired CO2 collected for 6 hr, suggesting that the 14C in the epoxy ring was intact.


Multicorrelation analyses of toxic and antigenic components of DPT and DP vaccines produced in Japan.

The combined diphtheria-pertussis-tetanus and combined diphtheria-pertussis vaccines, of which the contents of endotoxin, LPF and HSF were reported in the previous paper, have been tested for the respective potencies of pertussis, diphtheria and tetanus components. In order to obtain some information concerning possible adjuvanticities of the toxic components of pertussis cells, relationship between respective toxic activities and the potencies of the respective antigenic components were tested by applying the multiple regression analyses. The results showed that the effects of the toxic components were fairly complicated. Within the ranges of contents of the respective toxic components of the vaccines used in the present experiment, LPF appeared to enhance all the three potencies as the content increased, while the effects of endotoxin and HSF were either enhancing or suppressive depending on the antigenic components.


Augmentation of delayed-type hypersensitivity and resistance against allogeneic or syngeneic methylcholanthrene-induced tumors in mice preimmunized with the tumor extracts.

Delayed-type hypersensitivity (DTH) responses against methylcholanthrene-induced fibrosarcomas in C3H/He and BDF1 mice were developed in BDF1 mice by sc injection of the respective mitomycin C-treated tumor cells. The DTH responses to the allogeneic and the syngeneic tumor cells were accelerated and enhanced tumor-specifically by priming 7 days previously with KCl extracts of the respective tumors. The ability in the mice primed with the tumor extracts enhancing the DTH response against the tumor cells could be transferred to recipient mice by the spleen cells, but not by the T-cell-depleted spleen cells. Rejection of allogeneic tumor was accelerated under the development of accelerated and enhanced DTH response against the allogeneic tumor antigens. Moreover, resistance to syngeneic tumor growth increased significantly with the development of accelerated and enhanced DTH response against the syngeneic tumor antigens. Thus, the augmentation of DTH response by preimmunization with tumor extracts was accompanied by the increased resistance to tumor growth, suggesting that T cells involved in the augmentation of tumor-specific DTH response play some role in increasing the resistance to tumor growth.


[Disappearance and reversal of depth in stereopsis (author's transl)].

Four experiments were conducted to examine the transition of depth selection by mutual inhibition among disparities. (a) With random-dot stereograms, the time required for stereoscopic depth disappearance under fixation was found to be an inverse function of the magnitude of disparity. (b) Measurement of the cumulative time for perception of ambiguous stereograms showed that the depth was perceived longer with relatively smaller disparity. (c) The perception of stereoscopic depth under fixation was more stable in random-dot target than in ambiguous target. (d) Stereoscopic depth reversal occurred more frequently under fixation than in free observation. These results suggest that the whole range of disparities might be detected simultaneously and that the inhibition mechanism might have some important role in stereopsis.


[Punished behavior in hippocampectomized rats (author's transl)].

Hippocampectomized rats were compared with the controls on punishment training in a L-shaped alley. Five measures of running time were taken; startbox latency A (from the placement in the startbox to the door), startbox latency B(from the door to the runway), runway running time, goalbox entry time and goalbox time. The postshock running time of controls was significantly longer than the preshock one in all measures. Hippocampals showed no hesitation in both startbox latencies, but did show hesitation in the other three measures just as the controls did. The controls spent a long time in the startbox from which they were unable to see outside stimuli until reaching the startbox door. These results support the spatial cognition theory of hippocampal function rather than the distant cue theory.


The pathophysiology and surgical management of portal hypertension.

1. A classification of portal hypertension is given which relates better to the disordered pathophysiology than the older anatomical classification. 2. A review of the various procedural approaches to solution of the problems associated with portal hypertension has been given. 3. Based on our existing knowledge, a suggested approach to portal hypertension is described in which the various procedures and various types of shunts suggested over the years may be selectively applied to the appropriate patients.


The surgical management of malignant lesions of the pancreas.

Over the past 20 years, carcinoma of the pancreas has increased in incidence by 20 percent. In spite of this marked increase, there continues to be controversy in some areas regarding the proper surgical management of ductal lesions. To date, there have been 122 total pancreatectomies performed at the Mayo Clinic; 63 of these were performed for carcinoma of the pancreas. Among 21 patients treated by total pancreatectomy since 1975, there was one operative death for an overall operative mortality of 4.7 percent. During the same period of time, when 34 patients underwent partial pancreatectomy of the Whipple type, there were three postoperative deaths with an operative mortality of 8.8 percent. The three patients that expired following performance of the Whipple procedure did so as a result of leakage at the anastomosis between the pancreas and the jejunum. Endocrine and exocrine pancreatectomy management have not been a particular problem. Conclusions can be drawn that total pancreatectomy for carcinoma of the pancreas is an acceptable procedure.


Effect of noncoronary blood flow upon myocardial mitochondrial function during hypothermic anoxic arrest.

Mongrel dogs (23) were subjected to the experimental study characterizing the effect of noncoronary blood flow upon myocardial mitochondrial respiration. Anoxic arrest for 60 minutes was obtained by cross-clamping of the aorta under hypothermic cardiopulmonary bypass, and heart was reperfused for 10 minutes, then heart was excised to obtain the endocardium, epicardium of the left ventricle and ventricular septum for study of mitochondrial function and myocardial blood flow. Myocardial blood flow was measured with carbonized plastic tracer 46Sc during cross-clamping of the aorta. Noncoronary blood flow showed equal distribution in the left ventricle and septum with flow of 0.16 +/- 0.23 ml/ min/100g (endocardium). Mitochondrial respiratory function following 60 minutes of hypothermic anoxic arrest at 20 degrees C recovered to normal level, and also no correlation was demonstrated between noncoronary blood flow during cross-clamping of the aorta and mitochondrial respiratory function. It was concluded that noncoronary blood flow was negligible with respect to the oxygen demand at 20 degrees C of myocardial temperature, and that noncoronary blood flow during cross-clamping of the aorta was not correlated to mitochondrial protection from ischemia.


In vitro sensitivity test of stomach cancer tissues by the use of metal grid method.

Ninety-nine specimens obtained from 53 patients with stomach cancer were cultured for about 3 days by means of the stainless steel grid method. In vitro effects of antitumor drugs on the cancer cells were evaluated autoradiographically or biochemically using a liquid scintillation counter to measure the uptake of 3H-thymidine. The radioactivity of the labeled tumor cells of both control fragments and fragments affected by drugs varied greatly among individual tumors. Therefore, the in vitro efficacy of antitumor drugs was represented as a comparison with that of control fragments. Positive correlation between in vitro tests and the clinical effects of antitumor drugs was observed in the specimens of 18 cases.