Determining endotoxin content of ground beef by the Limulus amoebocyte lysate test as a rapid indicator of microbial quality. Eighty-four samples of ground beef were placed into five half-log cycle groups based upon aerobic plate count (APC) results. Endotoxins were determined by the Limulus amoebocyte lysate test (LAL), and gram-negative viable counts were determined by a violet red bile agar overlay method. Ten samples with a log of APC of less than 5.50 had an APC mean of less than 5.24 and mean endotoxin content by the LAL of 51 ng/g. The 15 samples with APCs between a log of 5.50 and 5.99 had an APC mean of 5.79/g and an endotoxin mean of 103.8 ng/g. Twenty-eight samples had APCs between a log of 6.00 and 6.49 with a mean of 5.28/g and an endotoxin mean of 1106.4 ng/g. The 20 samples with APCs between a log of 6.50 and 7.00 had a mean of 6.77/g and an endotoxin mean of 5067.6 ng/g, while 11 samples had a log of APCs of greater than 7.00 with a mean of 7.53 and an endotoxin mean of 7,472 ng/g. Correlation of half-log cycle mean APC and violet red bile agar counts with mean endotoxin content were both highly significant, indicating that LAL-determined endotoxin content can be used to make a rapid approximation of viable plate counts. Because results can be obtained by LAL in 1 h, the finding of low levels of endotoxins can be taken to indicate low-count meat. The use of additional tests of microbial quality may be necessary when high endotoxin levels are found because the LAL detects both viable and nonviable cells.


Sampling culturable heterotrophs from microcosms: a statistical analysis. The contributions of different sources of error in sampling mixed and unmixed bacterial microcosms were evaluated by using analysis of variance. Culturable heterotrophic bacteria from a turbid freshwater impoundment were sampled from 9-liter tanks that were unagitated or mixed with magnetic stirrers or pumps and from dilution bottles that were unagitated or agitated with a mechanical shaker. Axenic cultures of Enterobacter aerogenes were also sampled from manually shaken test tubes. In both agitated and unagitated tanks and in unagitated dilution bottles, dilutions made from the same sampling pipette were significantly different, showing a clumping of bacteria on the scale of millimeters. Also, microcosms within a single experiment differed from one another by a large margin. Dilution mean squares and tank or bottle mean squares were homogeneous for all types of tanks and unagitated bottles, indicating that the gentle mixing provided by pumps and stir bars did not reduce either millimeter scale or intermicrocosm variability over what prevailed in unagitated microcosms. By contrast, the vigorously shaken bottles and test tubes showed no millimeter scale variability. Intermicrocosm variability was undetectable in test tubes and two orders of magnitude less in shaken bottles than in unshaken bottles. When these facts are coupled with the inherent statistical advantage of replicating large rather than small experimental units, it is concluded that sampling error in the enumeration of aquatic bacteria in microcosms will be reduced by using numerous, small, violently agitated microcosms with a minimum of subsampling per microcosm.


Statistical procedure for evaluating the sensitivity of Limulus amoebocyte lysate by using a reference lysate. A study was designed to estimate variability of the Limulus amoebocyte lysate test by comparing a reference lysate against itself. Three technicians performed parallel tests, i.e., titrated side by side, the contents of two vials of reference lysate on 4 different days using 24 vials of the United States reference lysate and 12 vials of the United States reference endotoxin. Each parallel test was replicated three times. From the sensitivity endpoints, ratios were calculated for each parallel test. These ratios were converted to the logarithm for estimating variability among technicians and among vials of endotoxin. By using the overall variability of log ratios, a statistical procedure was developed to evaluate the sensitivity of each lot of licensed lysate submitted to the Bureau of Biologics for release.


Plasmids in Escherichia coli controlling citrate-utilizing ability. The citrate-utilizing ability of 19 out of 22 citrate-positive Escherichia coli strains isolated from pig sewage was transferred via conjugation to E. coli K-12. The conjugal transfer of citrate-utilizing (Cit) abilities was thermosensitive and concurrent with transfer of drug resistance. Weakly citrate-positive colonies were readily obtained in conjugation experiments. Their Cit characters could be transmitted to the other E. coli strains at a similar frequency in the retransfer experiments, and the transconjugants obtained still showed same characteristic growth on Simmons citrate agar plates. The 19 thermosensitive plasmids conferring citrate utilization and drug resistance were Fi-, and 16 of these plasmids belonged to incompatibility group H1. However, occasionally two conjugative plasmids (pOH3122-1 and pOH3124-1) carrying only the citrate utilization were also obtained in the conjugation experiments, and they were Fi+ and compatible with 19 reference R plasmids. In the two citrate-positive E. coli strains, it was suggested that the conjugative Cit plasmid showing Fi+ character and the more thermosensitive H1 plasmid conferring both the Cit character and drug resistance coexisted in the strain. The characterization of citrate utilization plasmids derived from pig farm sewage is discussed.


[Molecular aspects of carcinogenesis (author's transl)]. The mechanism of carcinogenesis is not yet understood. There is increasing evidence justifying the assumption that an unifying concept of carcinogenesis should be possible at the molecular level. New insights into the molecular mechanism of carcinogenesis were mainly obtained in studies on both chemical and viral carcinogenesis. In the present paper, selected results of these studies are reviewed. It is concluded that the interaction of different carcinogenic agents with the cellular DNA results in alterations of DNA. Only some of these alterations, however, seem to be relevant to carcinogenesis. Alterations of DNA can be caused by reaction of electrophilic agents with DNA constituents, by increased infidelity of DNA replication, by integration of viral genomes or by recombination events involving integrated proviruses.


[The significance of the enzyme histochemistry in the diagnosis of malignant non-Hodgkin's lymphomas (author's transl)]. The significance of histo- and cytochemical enzyme investigations in the diagnosis of malignant Non-Hodgkin's lymphomas (NHL) is evaluated. Histochemical enzyme methods complete the morphological diagnosis if some general principles of diagnostic histochemistry are observed. This is particularly true for the diagnosis of the hairy cell leukemia, the T-lymphoblastoma and the histiocytic reticulosarcoma.


Development of pancreatic endocrine cells in the rat fetus. Prenatal development of rat pancreatic endocrine cells was investigated by the immunoperoxidase technique and the following results were obtained: 1) Glucagon immunoreactive cells are first endocrine element of the pancreas appearing already on day 11 of gestation, when the dorsal pancreatic bud is still quite small and the ventral pancreatic primordium is hardly swollen out. Most of the glucagon reactive cells are located in the epithelium of the foregut and the dorsal pancreatic bud but a few of them are attached to the base of the epithelium from the outside. 2) Insulin and pancreatic polypeptide (PP) immunoreactive cells are first demonstrable in small cell clusters budding from the exocrine tubules on day 14, whereas somatostatin and gastrin reactive cells on day 17 and 18, respectively. These findings support the hypothesis that the majority of pancreatic endocrine cells is derived from the epithelium of the foregut. 3) PP reactive cells, appearing first on day 14, assume gradually a peripheral position in the growing islet of Langerhans. Immediately before birth they attain the population and flattened cell shape comparable to those in adult pancreas. Their counterpart in the duodenum is found as open type basal-granulated cells in the rat fetus. 4) Noteworthily, glucagon-like immunoreactivity is found in some neuron-like cells of the Auerbach's plexus between the muscle layers of the duodenum on day 19.


In vitro drug resistance of salmonellae in Singapore. Strains of Salmonella typhi isolated in Singapore were susceptible to chloramphenicol (except from two imported cases) and most of them also generally susceptible to ampicillin and septrin. Drug-resistant strains of Salmonella typhimurium appeared in 1971 and caused out-breaks among young children. However, decreasing drug-resistance of S. typhimurium was recorded in recent years following a drop in the number of cases. Other Salmonella serovers were found relatively susceptible to the antibiotics tested.


Concanavalin A surface receptors and cytoplasmic actin in cell adhesion. A double immunofluorescence staining technique to locate concanavalin A (Con A) surface receptors and cytoplasmic actin in the same cell was applied to monolayer cultures of rat foetal fibroblasts during cell detachment induced by trypsin and during cell attachment to glass substratum. Con A receptors were demonstrated by fluorescein-isothiocyanate-labelled Con A (FITC-Con A) and actin by specific anti-actin antibody (AAA) traced with rhodamine-labelled goat anti-human globulin (R-AHG). Untreated, control cells had an elongated shape, Con A receptors restricted to cell margins and prominent actin filaments. After 2 min treatment with 0.001% trypsin the cells became angular with Con A receptors in clusters and actin in a diffuse or aggreagate staining pattern. Progressive cell rounding followed and this was accompanied by the development of long, thin, arborized cell processes, studded with Con A receptors and containing fine actin filaments. Complete cell rounding preceded cell detachment. The sites of detached cells were marked by fine aggregates containing Con A receptors and actin. In cells attaching to a glass substratum, actin was present in a diffusely stained or aggregate pattern in round cells, in filaments restricted to cell margins in partially spread cells and in numerous filaments in fully spread cells. Con A receptors were present in clusters in round cells, in clusters or caps in partially spread cells and in cell margins in fully spread cells. Binding of FITC-Con A to partially spread cells resulted in dissolution of the few, newly formed, actin filaments. We believe our observations are consistent with the idea that actin filaments, formed during cell attachment, contribute towards the maintenance of cell adhesion by helping in the preservation of cell shape and by anchorage of Con A receptors at points of cell attachment to the substratum.


Laboratory diagnosis of bovine tuberculosis. Lesions of suspected bovine tuberculosis were examined by culture, histopathology and auramine-O (AO) stained smears and the findings correlated with field aspects of the disease. Of 642 lesions considered to be tuberculous, 62.0% yielded M. bovis and 4.5% other mycobacteria (OM). M. bovis and OM were recovered also from 0.6% and 3.6% respectively of 165 cattle which gave tuberculin reactions but had no visible lesions at slaughter. Of 262 lesions in which a histopathological diagnosis other than tuberculosis was made, 1.5% and 3.0% yielded M. bovis and OM respectively. All OM isolates tested belonged to the Mycobacterium-avium-intracellulare-scrofulaceum (MAIS) complex with a predominance of serotype 2. A good relationship was found between the recovery of mycobacteria and histopathology but examination of smears revealed 22.0% apparent false negatives. Apparent false negative culture results were also reported for 35.8% of lesions positive on histopathology and smear examination. The majority of herds yielding M. bovis contained reactors to the tuberculin test and many of these had lesions of tuberculosis. In contrast, herds yielding OM seldom contained reactors to the tuberculin test and rarely reactors with tuberculous lesions. The thoracic cavity was the main site of lesions from infections by M. bovis and OM.


Effect of environmental temperature on susceptibility of young chickens to Salmonella typhimurium. Day-old chickens kept in a cold environment (18 degrees to 22 degrees C) were more susceptible to a low and moderate challenge of Salmonella typhimurium than chickens similarly challenged and kept in a warm environment (32 degrees to 36 degrees C). Cold stress at 10 degrees C for 24 h when applied to 12-day-old chickens effectively increased the number of birds shedding organisms. However a similar cold stress on 20-day-old chickens resulted in a less dramatic increase in the number of birds shedding organisms. Of the 60 birds previously challenged with S. typhimurium and then subjected to cold stress, 16 birds recommenced shedding and 7 birds with no previous history of shedding began to shed organisms.


Vaccination of piglets against Escherichia coli enteritis. An immunisation procedure involving intraperitoneal followed by oral administration of a killed Escherichia coli vaccine was used to reduce losses associated with colibacillosis in young pigs reared under commercial conditions. Mortality was reduced in both the preweaning and postweaning period. The mortality rate among vaccinated piglets was half that of unvaccinated controls and by 3 weeks after weaning the total weight of pigs produced was 33% higher in the vaccinated group than in the control group.


Long-term study of oxdralazine in hypertensive patients. 56 moderate and severe hypertensive patients entered an open long-term trial aimed at evaluating the efficacy and tolerability of a combined treatment consisting of 3-hydrazino-6-[N,N-bis-(2-hydroxyethyl)amino]pyridazine (oxdralazine, L 6150), propranolol and chlorthalidone. The mean basal blood pressure was 186.9/111.8 mmHg: after one month of treatment, the mean value was 149.7/95.5 mmHg (p less than 0.01). The heart rate was practically unaffected by treatment, the mean value changing from 75.4 b.p.m. (basal) to 73.3 b.p.m. (one month). The significant reduction in the blood pressure observed at the end of the first month remained unchanged in the following months of therapy, and only minor variations occurred. The combined treatment was well tolerated. Five patients were withdrawn from the trial during the first month; three of them because of side-effects and two of them for personal reasons.


[Electron microscopic and microbiological studies on the activity of mezlocillin against Escherichia coli (author's transl)]. Investigations by electron microscopy on thin sections and negative-stained preparations were carried out to show the effect of 6-[(R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido]-penicillanic acid sodium salt (mezlocillin, Baypen) on the ultrastructures of gramnegative bacteria (Escherichia coli C 165). Distinct alterations of the bacteria were to be seen depending on concentration and cultivation time. The outer wall of the bacterial cell became irregular. Locally it detached from the plasma membrane. The normal density of the cytoplasmic structures decreased quickly. Owing to their inability to form septa the bacteria grew as extended filaments. These mezlocillin-treated cells could form filaments of up to 100 times the length of normal untreated bacteria. Those bacterial filaments, however, were unable to form colonies as was shown by survival curves carried out in parallel. The filaments, therefore, were irreversibly damaged bacterial forms.


[Results of a multicentric clinical study of mezlocillin in Japan (author's transl)]. In a clinical trial including 701 patients 6-((R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-1-carboxamido]-2-phenyl-acetamido)-penicillanic acid sodium salt (mezlocillin, Baypen) proved effective in the treatment of severe infections. It showed a particularly high efficacy in septicaemia and endocarditis and in purulent meningitis. In cases unresponsive to previous treatment with other antibiotics mezlocillin was effective in about two-thirds of the cases. Side effects were slight and of the same nature as those accompanying other penicillins.


Germination of non-irradiated and gamma-irradiated fungal spores on water agar and natural soil. Germination and germ-tube lengths of non-irradiated and gamma-irradiated spores of eight soil fungi were studied on water agar and on natural soil. Non-irradiated spores showed variable degrees of sensitivity to the fungistatic factor in natural soil. Lower doses of gamma-irradiation (5 K rad) produced stimulus to germination and germ-tube lengths without losing the fungistatic properties of the fungi concerned. Germination at the rest of treatments (100 and 500 K rad) showed significant inhibition, the magnitude of which is a function of the dose used. Germ-tube lengths did not follow the same pattern. The results are discussed in view of the nutritional theory of fungistasis.


On the transformation of coliform types, proper and irregular forms. Physiological changes have been induced in strains of E. coli I and its irregular forms, using a basal synthetic medium and addition of sodium pyruvate, or glucose, or yeast extract, or peptone. Changes in biochemical patterns, using the IMViC reactions plus growth at 44.5 degrees C, were induced, thus brining transformations between the regular types and irregular forms. Ultra-violet irradiation of E. coli I for 90 seconds may turn it into a methyl-red negative strain, while education of E. coli III for producing acid and gas at 44.5 degrees C was possible. Thus, E. coli I could be transformed to give the same reactions as the untyped form 1 of Geldreich (1966) through U.V. irradiation, while E. coli III may give the biochemical reactions of E. coli I via heat-resistance and training in MacConkey broth. The relationship between regular and irregular coliforms was also verified by phage typing. The results obtained were discussed both qualitatively, using "lytic activity", and quantitatively in terms of "efficiency of plating", especially with sewage-derived phages and irregular coliforms.


Primary lymphomas of bone (so-called ("Parker and Jackson's reticulum cell sarcoma") : histological review of 75 cases according to the new classifications of non Hodgkin's lymphomas. 75 cases of primary lymphomas of bone ("reticulum cell sarcoma" described by Parker and Jackson) were reviewed. Biopsy specimens were classified according to several histological classifications, including Lukes' classification for nodal lymphomas. Among 24 tumors described as "histiocytic lymphomas" in Rappaport's classification of 1966, only one case remains as "true histiocytic sarcoma". The others were reclassified as follows : 6 tumors with a predominance of large cleaved cells, 13 tumors with large non-cleaved cells, 2 as immunoblastic tumors and 2 as unclassifiable. In non Hodgkin's lymphomas, the histological diagnosis and the results of the initial staging have been shown to be greater aids in the prediction of survival than the primary site of the disease. Therefore, the clinical and therapeutical approach to the patients with lymphomas of bone must be modified to be similar to that of patients with nodal localisations of the disease.


The relationship between reproductive activity and blood calcium in the calcium-deficient hen. 1. The dependence of reproductive activity in the laying hen upon adequate calcium intake has been investigated. 2. The response of plasma luteinising hormone concentration in calcium-deficient, as compared with calcium-replete hens, to injections of luteinising hormone releasing hormone and progesterone suggested that the primary site of reproductive dysfunction is the hypothalamus rather than the pituitary gland. 3. Laying hens, when presented with a calcium-deficient diet, ceased to lay as the plasma ionised calcium concentration decreased to less than 1.0 mM, supporting the view that there is a threshold of blood calcium activity below which reproductive activity ceases. 4. Supplementation of the diet with calcium produced an immediate restoration of plasma ionised calcium concentration to normal, despite an interval of a few days before the plasma total calcium returned to normal and egg laying resumed.


The adverse effects of nicarbazin on reproductive activity in the hen. The effects of the anticoccidial agent nicarbazin on reproductive activity in the female fowl have been studied. 2. The drug had no effect on the plasma concentration of luteinising hormone, but treated hens showed a reduced hypothalamic sensitivity to exogenous progesterone, whilst the capacity of the pituitary to respond to luteinising hormone releasing hormone was unimpaired. 3. It is suggested that nicarbazin not only prevents yolk deposition within the ovary but also adversely affects the stimulatory function of the hypothalamus, possibly through an unsuitable hormonal environment.


Sensations from surgically transferred glabrous skin; central versus peripheral factors. In reinnervated skin transferred from the foot to the hand, sensory thresholds approach normal values--an observation not predicted by the theoretical relationship between innervation density and tactile acuity. Therefore, we suggest that innervation density is not the major factor determining tactile acuity. Rather, the hand region of the central somesthetic map may be specialized to provide a calibre of function unavailable to other regions.


Medical aspects of the preoperative management of aneurysms: a review. The aim of medical management of a patient with a recently ruptured intracranial aneurysm is to preserve residual brain function and prevent systemic complications. Surgery should be performed as soon as the patient is in good neurological condition. Most fatalities result from the destructive effects of the initial hemorrhage, but delayed ischemic neurologic deficit can result from vasospasm and rebleeding. Systemic complications of the brain damaged state result in a smaller proportion of deaths. Common medical problems are reviewed as well as their medical management. In particular, some special problems related to subarachnoid hemorrhage such as cerebral edema and herniations, rebleeding, and vasospasm are also considered. Major recent advances have been the introduction of antifibrinolytic therapy, the realization of the importance of maintaining blood volume and pressure, as well as general advances in respiratory care.


Effect of insulin-induced hypoglycemia on the serum concentrations of thyroxine, triiodothyronine and reverse triiodothyronine. The effect of insulin-induced hypoglycemia on serum thyroid hormone concentrations was studied in nine healthy individuals. Before, during and after the hypoglycemia blood samples were taken for measurement of the concentrations of glucose, thyroxine (T(4)), triiodothyronine (T(3)), reverse triiodothyronine (rT(3)), catecholamines and pituitary hormones.There was no change in the mean serum T(4) level (+/- the standard error of the mean) of 67 +/- 2 mug/l. However, the T(3) concentrations rose from a mean basal level of 1.86 +/- 0.06 mug/l to a mean peak of 2.51 +/- 0.21 mug/l (P < 0.01) at 45 minutes after the insulin injection, and the rT(3) concentrations fell from a mean basal level of 0.184 +/- 0.008 mug/l to a mean nadir of 0.171 +/- 0.022 mug/l (not a significant change). The mean peak epinephrine level was 545 +/- 103 ng/l and it occurred between 30 and 45 minutes after the insulin injection; the mean peak norepinephrine level was 584 +/- 114 ng/l and it occurred between 30 and 90 minutes after the injection. The growth hormone levels reached a mean peak of 26.1 +/- 4.8 mug/l and the plasma cortisol levels rose to 215 +/- 9 mug/l. The mean basal prolactin level was 8.5 +/- 0.9 mug/l; in five subjects there was a rise to a mean peak of 50.6 +/- 14.6 mug/l, whereas in the remaining four no significant increase occurred. No correlation was found between the changes in the serum T(3) concentration and any of the other factors studied.It was concluded that acute hypoglycemia is associated with a rapid increase in the serum T(3) concentration.


Proliferative hemorrhagic enteropathy in swine: an outbreak and review of the literature. An account is given of an acute outbreak of hemorrhagic enteric disease in 34 young adult boars of primarily Yorkshire breeding at a performance testing station in Western Canada. Two of the boars died. A diagnosis of proliferative hemorrhagic enteropathy was made on the basis of clinical signs, gross necropsy and histopathological lesions. Campylobacter-like bacteria were seen in the apical cytoplasm of epithelial cells of the ileal mucosa by Warthin-Faulkner stain on histological sections, but were not isolated on culture. Treatment with several drugs simultaneously was associated with termination of the outbreak, but the therapeutic effect was not controlled by leaving some animals untreated. Reported sequelae, such as porcine intestinal adenomatosis, were not present at necropsy or slaughter of pigs surviving the outbreak, but all were killed no longer than two months after the first case was observed. This outbreak is discussed in the context of previously reported hemorrhagic enteric conditions of pigs.


Differential effect of a microsomal deacetylase inhibition on the mutagenicity in Salmonella typhimurium of 2-acetylaminofluorene by liver homogenates of guinea pigs, mice and rats. The effect of paraoxon, a microsomal deacetylase inhibitor, on the mutant genicity of 2-acetylaminofluorene (AAF) by liver homogenates was compared between the AAF carcinogenesis-resistant guinea pigs and the susceptible mice and rats. The mutagenicity of AAF was mostly abolished by paraoxon, not only in the 3 kinds of untreated animals but also in guinea pigs treated with a combination of phenobarbital and 5,6-benzoflavone, whereas about 50% of the mutagenicity was resistant to paraoxon in treated mice and rats. We suggest that microsomal deacetylase activity is crucially involved in the mutagenic activation of AAF by guinea pig liver homogenates, while the enzyme activity other than the deacetylase activity is also important in the activation by liver homogenates from treated mice or rats.


Proteins and glycoproteins from human erythrocytes infected with Plasmodium falciparum. The purpose of this study was to determine whether metabolites of parasite origin are incorporated into the "knob-like" protrusions of membranes of erythrocytes infected with Plasmodium falciparum. Intraerythrocytic parasites were grown in culture and labelled metabolically with ((35)S)-methionine. Fractions enriched in membranes with knobs and fractions poor in knobs were prepared and analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Labelled components were visualized by autoradiography. Fractions enriched in membranes with knobs showed a consistent relative increase in a labelled band with an apparent relative molecular mass around 70-80 thousand. Membrane-enriched fractions were also tested for glycoprotein components using lectin-binding. No differences were observed between wheat germ agglutinin-binding components in membranes from normal and parasitized erythrocytes. On the other hand, membranes from parasitized erythrocytes showed distinct concanavalin A-binding components around 70-80 thousand relative molecular mass range. Further experiments are required to conclude whether the metabolically labelled protein component observed within the same relative molecular mass range is a membrane-associated glycoprotein identical to the concanavalin A-binding component.


Fractionation and characterization of Plasmodium falciparum antigens. In human malarial infections, the immunological importance of modified host cell components and of parasite antigens that may be released on spontaneous rupture of infected erythrocytes in vivo is not known. It is difficult, however, to analyse antigens extracted from entire parasitized red cells because of contamination with normal red cell constituents, notably haemoglobin. To overcome this problem, a simple chromatographic procedure has been developed that concentrates antigens and radiolabelled parasite products in a separate fraction from haemoglobin. Details are also given of investigations of two different kinds of antigen associated with Plasmodium falciparum. One antigen known to be immunogenic in man is shown to be associated with membranes. The other antigen is poorly immunogenic in man and other animals and is shown to be a soluble, rather stable, acidic protein.


Some aspects of serum requirements for continuous cultivation of Plasmodium falciparum. As possible substitutes or supplements for the human serum requirements for continuous cultures of Plasmodium falciparum several different animal sera have been tried, including: fresh fetal bovine, sheep, horse, adult bovine, newborn calf, swine, commercially supplied fresh-frozen or lyophilized human sera, fatty acid-free bovine or human serum albumin, red cell extract, Physiogel, and a serum substitute composed of Bacto-peptone, Yeastolate, Lactalbumin hydrolysate, polyvinylpyrrolidone, and bovine insulin. None were comparable to the human serum, nor did they exhibit any serum-sparing effect. Plasma containing citrate-phosphate-dextrose (CPD) was nearly as good as serum, but dialysed plasma or serum would not support cultures of P. falciparum. Fresh human serum varied from lot to lot in its culture supporting properties especially if the concentration of serum in the medium was reduced below 10%. At 10% concentrations, however, most sera supported the cultures very well.


In vitro propagation of Plasmodium falciparum for merozoite antigens. A system with continuous flow of medium that uses disposable tissue culture flasks for the in vitro cultivation of P. falciparum is described. This scaled-up modification of the Trager and Jensen method gives merozoite yields sufficient for experimental vaccine studies in animals. A method of increasing the yield of merozoites by decreasing the pH of the medium is presented.


Evidence for environmental modulation of gametocytogenesis in Plasmodium falciparum in continuous culture. With the introduction of continuous culture of Plasmodium falciparum it has become possible to study the factors involved in gametocyte production in vitro and thus eliminate the uncontrollable in vivo variables of the host. The authors have developed a method for measuring quantitatively the rate of production of gametocytes at any time in such cultures. The method is based on an estimation of the percentage of ring forms that develop into stage II gametocytes.Using this approach, it was found that dilution of cultures with fresh red blood cells so as to lower the parasitaemia led to rapid fall in the rate of conversion to gametocytes. The conversion rates subsequently rose again to levels typically in the order of 10% after several days of growth in the new culture. In the parental cultures from which the dilutions were made, conversion rates remained high at all times. This pattern was consistently observed in three different isolates of P. falciparum from Africa and the results indicate that the reduction of parasitaemia by addition of fresh cells was responsible for reducing production of gametocytes and that conditions associated with a period of growth in culture induced renewed gametocytogenesis. The authors conclude, therefore, that environmental conditions directly modulate the rate of gametocyte production by P. falciparum in culture.After 1(1/2) years in culture, parasites have retained their ability to produce gametocytes and the gametocytes to undergo exflagellation.


In vitro production and partial purification of Plasmodium falciparum antigen. A simple technique for achieving high yields of Plasmodium falciparum parasites on a continuous basis is described. The technique is applicable in any laboratory. The culture apparatus is also simple and inexpensive and allows multiple cultures to be run simultaneously. A total of approximately 1-2 x 10(9) parasites can be harvested per culture flask per week requiring the use of only 40.0 ml of culture medium (RPMI 1640), 5.0 ml of human sera, and 2.0 ml of outdated human whole blood. P. falciparum parasites (segmenters containing individual merozoites) are cultured in vitro and concentrated 10-15 fold through the use of discontinuous bovine serum albumin gradient centrifugation.Commercial saponin is purified on a Sephadex G-25 column. The haemolytic effect of purified saponin related to human red blood cell concentration is studied. Preliminary observations on the action of some synthetic detergents and enzymes on human erythrocytes are also reported. Purified saponin is used to lyse red blood cells infected with in vitro cultured P. falciparum for the preparation of merozoite antigen. Further purification of parasite material is carried out by sucrose density gradient centrifugation.


Soluble antigens released in vitro from erythrocytes infected with Plasmodium berghei. Rat erythrocytes parasitized with Plasmodium berghei were found to release soluble antigenic products in vitro. The supernatants obtained by centrifugation of the disrupted cells induced specific transformation of non-adherent spleen lymphocytes of convalescent rats and produced precipitation lines with antiplasmodial antibodies. In a preliminary experiment they also immunized young rats against a viable challenge of P. berghei. It is suggested that the soluble material could serve for the purification of an effective protective antigen.


Harvest of Plasmodium falciparum merozoites from continuous culture. Spontaneously released merozoites were harvested from cultures in which 42-90% of the erythrocytes had been infected with mature forms of Plasmodium falciparum at the start of incubation. The mature forms had been extracted from asynchronous cultures by the use of Ficoll and Plasmagel gradients. As the mature forms consisted of both trophozoites and schizonts, merozoites were released into the culture medium over a long period of time. The synchrony of merozoite release did not appear to be improved by prior exposure of parasites to sorbitol. Over this prolonged period of incubation, the yield of merozoites was disappointingly low in cultures containing 2.5% of erythrocytes. At erythrocyte concentrations of 0.01-0.25%, 3-10 times more merozoites were released into the medium; 0.4-2.3 merozoites per initial mature form were harvested over a 15-19-hour period. In addition to merozoites, contents of the culture medium included intact erythrocytes, ghost cells, and other cellular fragments. Only intact erythrocytes were effectively removed from the medium by simple or Ficoll gradient centrifugation. Merozoite preparations that are free from host cellular material are important in the development of a human malaria vaccine.


Recent developments in enlarging the scale of production of Plasmodium falciparum in vitro. Several methods are described for the cultivation of Plasmodium falciparum. They include cultivation in (a) flow vessels that contain 12 ml of RBC suspension and are harvested three times a week, (b) a "tipper" that provides a similar yield, and (c) more recently, a large flat-bottomed vessel that holds 75 ml of suspension. Attempts are being made to develop techniques for synchronizing the cultures.


Lidoflazine in the management of angina pectoris. The effect of lidoflazine administration (120 mg t.i.d. for 9 weeks) on work tolerance (bicycle ergometer), frequency of anginal attacks, and nitroglycerin consumption was investigated in 28 male patients with stable angina pectoris in a combined single-blind/double-bline study. Lidoflazine increased work tolerance and reduced the frequency of anginal attacks and nitroglycerin consumption. The higher work tolerance level was reached at maximal heart rate and heart rate--systolic blood pressure product values similar to those before treatment. The values of these variables after 3 min of exercise at 60 W, however, were significantly lower after treatment with lidoflazine. These findings indicate that the heart is performing more economically during lidoflazine treatment. The improved work tolerance can probably be ascribed to lidoflazine and not to a training effect because of the significant reduction of this variable in the patients allocated to placebo as compared to those remaining on lidoflazine treatment. The side effects were generally slight. In one patient the prolongation of the QT interval due to lidoflazine resulted in rhythm disturbances.


Lidoflazine and physical training in the treatment of stable angina pectoris. An additive effect in the treatment of angina pectoris by combining lidoflazine and physical training was postulated. Twenty-four patients were randomly divided into placebo and drug groups and subsequently underwent a 6-month physical training program. The drug group had a significantly greater reduction in submaximal heart rate than the placebo group. Similar improvements in symptom-limited exercise capacity were observed in both groups. Resting and maximal exercise coronary sinus blood flow and left ventricular oxygen consumption were not significantly changed with training in either group. Physical training and lidoflazine appear to influence exercise tolerance in the same manner.


Microsurgical approach to the anterior cervical spine and spinal cord. When the cervical spine is affected by disease that is predominantly anterior, we prefer to take an anterior operative approach. Whether or not fusion is indicated depends largely upon the case. In general, we do not fuse the spine unless it is unstable. Widening of the spinal canal and the neural foramina can be done effectively by the anterior route, with or without anterior fusion. When the spine is unstable from disease affecting the vertebral bodies, we routinely fuse with bone graft or provide mechanical stability with a vertebral body prosthesis if life expectancy is short.


Metabolic disorders in myocardial infarction. Changes in blood serum zinc, growth hormone, insulin and glucose concentration in patients with acute myocardial infarction. The authors examined the concentrations of zinc, HGH, insulin and glucose in blood serum of 40 patients with acute myocardial infarction. Zinc and HGH concentrations in blood serum correlated well with changes in the clinical state in these patients. The more severe the condition of the patient, the greater zinc concentration fall and HGH concentration rise in blood serum were noted. All the changes were statistically significant (P less than 0.001) in cases of severe clinical course and most distinct in cardiogenic shock. On the other hand, in MI of mild course, zinc concentration fall in blood serum was not accompanied by HGH rise. A positive correlation between the concentrations of zinc and HGH found in normal subjects became negative in patients with acute MI of severe course. The changes of zinc and HGH concentrations seem to be due to the non-specific effect of acute myocardial stress, particularly enhanced in MI with severe course and cardiogenic shock, Observation of the intensity of changes may have prognostic significance due to the fact that they were most distinctive in MI of most severe course.


Reconstitution of the solubilized insulin receptor in phospholipid vesicles. The insulin receptor was solubilized from turkey erythrocyte membranes by extraction with 1% beta-octylglucopyranoside. Insulin binding was enhanced when the solubilized material was reconstituted in phospholipid vesicles. The affinity of the reconstituted vesicles for various insulins was similar to that of the intact membranes: porcine insulin greater than proinsulin greater than desoctapeptide insulin. A curvilinear Scatchard plot was obtained for insulin binding to the reconstituted system at 15 degrees C. A high affinity association constant of 1.4 x 10(9) M-1 was obtained from the Scatchard plot. This is a four-fold increase over the value for the turkey erythrocyte membrane, which contains more highly saturated phospholipids. This suggests that the insulin receptor may be sensitive to the lipid composition of the membranes in which it is embedded.


[The effects of electric stress on pancreatic B cell function in rats fed a high fat diet (I). Glucose tolerance and glucose-induced insulin release from the perfused pancreas (author's transl)]. The synergistic effects of dietary obesity produced by the feeding of a high fat diet and stress induced by electric shocks on glucose tolerance and glucose-induced insulin release from the perfused pancreas were investigated. Male Wistar rats weighing 90 approximately 100 g were fed ad libitum for 12 weeks either a control (50% Starch; C) or a high fat diet (40% Butter; F). Some of the rats on both diets received 100 electric shocks of 1 sec. duration in the stress session for 1 hour per day for the last 3 weeks of the experimental period. Low stress (LS) groups were shocked at a fixed time (Inter Shock Interval: 36 sec.). High stress (HS) groups were shocked at random (ISI: mean = 36 sec, 9 approximately 108 sec. variable). Non-stress (NS) groups were not given any shocks. Rats were killed at 24 hours after the final stress session. Under NS conditions, rats in the F-NS group gained a significant amount of weight and had normal levels of fasting plasma glucose and insulin but an impaired glucose tolerance (k = 3.49). Insulin release from the perfused pancreas in the F-NS group showed a delay in the initiation of release by the stimulation of glucose (16.7 mM), but the total amounts of insulin released did not differ from that in the C-NS group. On the other hand, the levels of plasma 11-OHCS in the fed state were much more highly elevated in the HS group than in the LS group, which was not influenced by the high fat diet. The fasting levels of plasma glucose in the F-HS group (121 +/- 7 mg/100 microliter) were significantly higher than those in the C-HS group (101 +/- 7 mg/100 microliter) in spite of a normal insulin concentration in plasma. In contrast to the normal glucose tolerance in the C-HS group (k = 5.14), glucose tolerance in the F-HS group (k = 3.04) was impaired. Insulin release from the perfused pancreas in response to glucose in both diet group was not significantly altered under LS conditions. In the C-HS group, however, the total amount of insulin released in the second phase was enhanced to 165% of that in the C-NC group. Conversely, in the F-HS group the total amount of insulin released in the first phase was significantly decreased to 40% of that in the F-NS group. These findings indicate that the elevation of plasma 11-OHCS levels provoked by shocks at random rather than in a fixed time schedule is caused by the difficulty in predicting shocks, and a chronic stress induced by electric shocks at random further impairs glucose tolerance and suppresses glucose-induced insulin release in rats fed a high fat diet.


[Study on enzyme immunoassays for the measurement of thyroglobulin and anti-thyroglobulin autoantibody in human serum (author's transl)]. Enzyme-linked sandwich immunoassays for the measurement of thyroglobulin and anti-thyroglobulin autoantibody in human serum using silicone rod and beta-D-galactosidase were studied. These methods showed excellent results in specificity, sensitivity, precision and clinical application. 1) A method using silicone rod coated with rabbit (anti-human thyroglobulin) immunoglobulin G and rabbit (anti-human thyroglobulin) monovalent fragment of immunoglobulin G (Fab') conjugated with beta-D-galactosidase was developed for the measurement of circulating thyroglobulin. The sensitivity of the assay with as little as 2 microliter of serum was 10.7 amoles/tube corresponding to 3.5 ng/ml of serum, which was equal to or rather higher than that of radioimmunoassay. The correlation coefficient between values determined by the present assay and a double-antibody radioimmunoassay was 0.99 (n = 63, p less than 0.001). Circulating thyroglobulin was detectable in 90% of 146 normal subjects, the concentration being 13.3 +/- 10.3 ng/ml (mean +/- S.D.). Interference of anti-thyroglobulin autoantibody with the assay for thyroglobulin was smaller than that in radioimmunoassay. 2) Another method using human thyroglobulin conjugated with beta-D-galactosidase and silicone rod coated with human thyroglobulin was developed for the measurement of circulating (anti-human thyroglobulin) autoantibody. The sensitivity of the assay with as little as 5 microliter of serum was 7 fmoles/tube corresponding to 220 ng/ml of serum, which was equal to or rather higher than that of radioimmunoassay. The highly significant correlation was observed between the concentrations of anti-thyroglobulin autoantibody determined by the present assay and a radioimmunoassay (r = 0.80, n = 74, p less than 0.001) and also between those by the present assay and those by tanned red cell hemagglutination (r = 0.78, n = 199, p less than 0.001). No effect of thyroglobulin on the present assay was observed unless the ratio of the amount of thyroglobulin to that of (anti-human thyroglobulin) immunoglobulin G was higher than a tenth.


Solid phase immunosorbent technique (SPIT) for the demonstration of specific IGM antibodies by hemagglutination inhibition and passive hemagglutination. The adsorption of protein to glass and plastic has been well documented. It has also been demonstrated that the absorbed antigen or antibodies retain their activity in as much as they are able to react with a specific antibody respective specific antigen(1). Furthermore the adsorbed antignene or antibody complex may also react with a later added antigen. Anti IgM is adsorbed to the solid phase and will retain IgM from the patient's serum. If the patient's serum contains specific IgM this will react with the specific antignene. If the antigen is a hemagglutinin it will combine with the specific IgM and therefore added erythrocytes will not hemagglutinate. If the antigen is coated on erythrocytes the coated erythrocytes will combine with the specific IgM and hemagglutination will take place. The reaction is schematically demonstrated in Fig. 1.


A prospect on cancer immunology.

A prospect on cancer immunology. There are several factors involved in studying cancer immunology. For convenience, those factors can be consolidated into two. Firstly, no definite tumor-specific or -associated antigen has been ascertained as yet, except for certain types of tumor. Secondly, there is no definite pattern of immune response of the host against weak antigenic tumor cells. Nobody knows as to what is the nature of the tumor-specific antigen even if it exists, and nobody knows the escape mechanism of tumor cells from the immune defence of the host. There have been a number of approaches for cancer immunotherapy, but so far there has been no definite answer as to whether immunotherapy is a promising approach for cancer treatment. In this review, cancer immunology is divided into three separate subjects. The first of these is tumor antigen; the second, the immune response against tumor antigen; and the third, methods of attacking tumor cells by immunological means including how to increase the antigenicity of tumor cells (xenogenization), and how to increase the immune response of the host (immunotherapy).


Glycophorin A as a cell surface marker of early erythroid differentiation in acute leukemia. We show here that the leukemic blast cells from three patients of a total of 15 subsequently diagnosed as having acute leukemia express on their surface the major red cell sialoglycoprotein, glycophorin A (GP-A). This was demonstrated (1) by indirect immunofluorescence using rabbit anti GP-A anti-serum and (2) by immune precipitation of GP-A from surface radiolabelled leukemic cells. Since GP-A is exclusively present on erythroid cells and their precursors, these findings indicate that a higher proportion of the blast leukemias than previously recognized show features of early erythroid differentiation.


An assay for estradiol preceptors using a staphylococcal protein-A--estradiol antibody adsorbent. Antiestradiol antisera were raised in rabbits by immunization with a steroid-protein conjugate Whole antisera and purified antiestradiol antibodies were reacted with heat-killed and formalin-fixed bacteria from the protein-A-bearing strain of Staphilococcus aureus (Cowan I, NCTC 8530). The bacterial immunoadsorbent was used to estimate the estrogen-receptor proteins contained in several specimens from human breast carcinoma. Since the affinity constant for estradiol of antiestradiol antibodies is about 10(8) M-1 while those for breast estrogen-receptor complexes are about 10(9) to 10(10) M-1, it is possible to remove the free steroid from a reaction mixture containing tissue cytosol, radio-labelled estradiol, and the antiestradiol bacterial adsorbent by pelleting the bacteria at low-speed centrifugation. The radiolabelled estradiol molecules bound to their receptors remain in the supernatant and can be easily counted by liquid scintillation counting. The results obtained by this technique on specimens of human breast carcinomas compared more than satisfactorily with those obtained, on separated aliquots from the same specmens, by the dextran-coated charcoal method, according to the EORTC group (1973). The versatility, specificity and stability of the antiestradiol bacterial immunoadsorbent recommend its use, rather than that of charcoal-coated dextran and other non-specific steroid adsorbents, in the mass screening of patients with breast carcinoma which could be amenable to hormonal therapy.


Effect of salt solutions on the radiosensitivity of mammalian cells as a function of the state of adhesion and the water structure. The radiation isodose survival curve of attached Chinese hamster (V79) cells, subjected to a wide concentration range of salt or sucrose solutions, is characterized by two maxima separated by a minimum. Cells are radioprotected at the maxima (high and low hypertonic salt concentrations) while they are radiosensitized at the minimum (intermediate hypertonic salt concentrations). Both cations and anions can alter the cellular radiosensitivity above and beyond the (osmotic) effect observed for cells treated with sucrose solutions. However, the basic curve shape, except in the case of sulphate salts, remains the same. When these experiments are repeated with single cells in suspension, the isodose survival curve is quite different in that high salt concentrations (greater than 0.9 M) do not protect cells in suspension unlike the case with attached cells. The curve shape is also altered in that the second maximum is absent with many salt solutions. If multicellular spheroids are used for these experiments, the data resemble those for single cell suspensions rather than for attached cells. The radiation survival data for cells in suspension in salt solutions correlate with water proton spin-lattice relaxation time (T1) and, in hypo- and iso-tonic solutions, with cell volume.


Comparative tumor-inhibitory and anti-bacterial activity of soluble and particulate glucan. A soluble fraction of particulate glucan was prepared and evaluated for its anti-tumor and anti-bacterial activity. Thin-layer chromatographic analysis indicated that the soluble preparation was composed of a variety of polyglucoses. Intravenous administration of soluble or particulate glucan resulted in significant reductions in the growth of a syngeneic anaplastic mammary carcinoma and melanoma B16. Survival data demonstrated that intravenous administration of soluble or particulate glucan prolonged survival of A/J and C57BL/6J mice with subcutaneous tumor implants. As regards to bacterial infections, soluble and particulate glucan decreased renal necrosis in S. aureus challenged mice as compared to control mice. Although the exact nature of the active soluble fraction(s) of glucan remains to be delineated, these studies demonstrate that a soluble glucan preparation exhibits significant anti-tumor and anti-staphylococcal activity. The active soluble fraction of particulate glucan may be preferable to particulate glucan in view of the inherent ease of parenteral administration.


Separation of dog brain renin-like activity from acid protease activity. A renin-like enzyme and acid protease (cathepsin) from whole and saline-pefused dog brains were separated by CM-cellulose chromatography with a linear NaCl gradient. Plasma renin and cathepsin were also separated using the same system. During the separation steps (in all the above cases) the specific activity of the brain renin-like enzyme was increased, while the specific activity of the brain cathepsin was decreased. Approximately a 70-fold increase in the specific activity of brain renin-like enzyme and a sixfold decrease in brain cathepsin specific activity was obtained from saline-perfused brain. The separation made it possible to study the pH optimum of the brain renin-like enzyme and acid protease. The brain renin-like enzyme showed optimal activity in the range of pH 6-7. Immunologically, the renin-like enzyme was distinctly different from dog kidney renin.


Modification of high LET radiation-induced damage and its repair in yeast by hypoxia. The lethal response of a diploid yeast strain BZ34 to densely ionizing radiations from the reaction 10B(n, alpha)7 Li was studied. The values for relative biological effectiveness (r.b.e.) and oxygen enhancement ratio (o.e.r.) for this radiation compare favourably with the data obtained with charged particles on the same strain of yeast. Recovery from potentially lethal damage was also studied by post-irradiation holding under non-nutrient conditions. In order to understand the role of oxygen in the recovery process, the investigation covered the following treatment regimens: (a) aerobic irradiation and aerobic holding (A-A), (b) aerobic irradiation and hypoxic holding (A-H), (c) hypoxic irradiation and hypoxic holding (H-H) and (d) hypoxic irradiation and aerobic holding (H-A). It has been found that the presence of oxygen is essential for recovery from the damage induced by both gamma rays and high linear energy transfer (LET) radiations. The extent of recovery was larger for gamma-induced damage than for damage induced by high LET radiation (alpha + 7Li) for the A-A condition. In the H-H condition, while only a slight recovery was seen for gamma-induced damage, it was totally absent for high LET damage. For the modality A-H, it was found that there is not recovery from the sparsely ionising gamma radiation-induced damage. The implications of these results for the treatment of malignant tumours by radiotherapy are briefly discussed.


Prolongation of the saralasin responsive state of two-kidney, one clip Goldblatt hypertension in the rat by the orally administered converting enzyme inhibitor captopril (SQ14,225). Two-kidney, one clip Goldblatt rats were treated with oral converting enzyme inhibitor captopril (SQ14,225) 6 mg/kg/day for 3 weeks after they had developed hypertension. Before treatment, systolic blood pressure rose from 143 to 202 mm Hg (p less than 0.05). Tail vein infusions of saralasin 10 microgram/kg/min in conscious rats reduced systolic blood pressure from 202 to 121 mm Hg (p less than 0.05) at 8 weeks after clipping the renal artery and before treatment with captopril. Chronic treatment with captopril for 3 additional weeks lowered blood pressure to 173 mm Hg (p less than 0.05). When saralasin infusion was repeated during treatment with captopril, blood pressure fell from 173 to 159 mm Hg (p less than 0.05). Blood pressure rose to 197 mm Hg within 4 days after captopril was discontinued and saralasin infusion 3 weeks after captopril (15 weeks after clipping the renal artery) again resulted in a dramatic fall in blood pressure from 197 to 142 mm Hg (p less than 0.05). Goldblatt rats who had not received captopril showed no blood pressure response to saralasin infusion at 12 weeks after renal artery clipping. The present study demonstrates that partial inhibition of the renin-angiotensin system with captopril results in a delay in the natural evolution of clip hypertension retarding the appearance of hypertension that is resistant to acute saralasin infusion.


Biochemical characterization of unidentified microaerophilic cocci isolated from heifer and dry-cow mastitis. Thirty-nine strains of gram-positive microaerophilic cocci isolated from cases of heifer and dry-cow mastitis were biochemically characterized with the API 50E and API-ZYM test kit systems, gas-liquid chromatography for analysis of end products of glucose metabolism, and anaerobic biochemical tests (L. V. Holdeman, E. P. Cato, and W. E. C. Moore, Anaerobe Laboratory Manual, Virginia Polytechnic Institute, Blacksburg, 1977). Strains were screened for production of a variety of extracellular enzymes on substrate-containing agar plates and for hemolysin and coagulase production. Antibiotic susceptibility and sensitivity tests were also performed. The microaerophilic cocci displayed homogeneity with respect to the majority of the biochemical tests used; i.e., greater than or equal to 90% of the strains were consistently positive or negative in any one test and probably represent one species. All produced deoxyribonuclease, ribonuclease, and hyaluronidase, and 92% were positive for chondroitin sulfatase. Catalase and coagulase tests were negative. Greening was observed on bovine blood agar. Acetic and succinic acids were produced by all strains as the only detectable products of glucose metabolism. The strains were susceptible to penicillin G, cefoxitin, doxycycline, and chloramphenicol and were resistant to clindamycin, novobiocin, and metronidazole. Their taxonomic position remains unclear.


Failure of serology in diagnosing chlamydial infections of the female genital tract. Chlamydia trachomatis was recoved from 20% (36/180) of women attending a venereal disease clinic. All infected women had chlamydial antibodies in their serum and cervical secretions. However, the background rates of chlamydial antibody in chlamydia-negative women were very high. Measurement of antibodies in serum (complement fixation or immunoglobulin G [IgG] and IgM by microimmunofluorescence) or cervical secretion (IgG, IgM, IgA or secretory IgA classes) did not result in predictive values of greater than 32%. It is concluded that the detection of chlamydial antibodies in serum or cervical secretions cannot be substituted for agent isolation in diagnosing these infections.


Vibrio alginolyticus infections in humans. Two clinical isolates of Vibrio alginolyticus from New Jersey are reported, one from a mixed stump infection and the other grown in pure culture from the conjunctival discharge of a man with conjunctivitis. The biochemical characteristics and antibiotic susceptibilities of these two isolates are presented. Human infections caused by V. alginolyticus are reviewed.


Evaluation of a biphasic medium for blood cultures. A study comparing the recovery of microorganisms from a transiently vented biphasic brain heart infusion medium bottle and a vacuum bottle containing tryptic soy broth demonstrated that growth was initially detected on the slant of the biphasic bottle or on a routine subculture of either broth in nearly 50% of instances. All organisms were isolated equally well in both bottles, with the exception of Staphylococcus aureus, which was isolated significantly more frequently from the biphasic bottle, and anaerobic bacteria, which were isolated significantly more frequently from the tryptic soy broth bottle.


Measure of immunoglobulin G-, M-, and A-specific titers against Legionella pneumophila and inhibition of titers against nonspecific, gram-negative bacterial antigens in the indirect immunofluorescence test for legionellosis. A crude extract of Escherichia coli O13:K92:H4 inhibited 97% of positive indirect immunofluorescence titers against a variety of gram-negative bacterial antigens while lowering Legionella pneumophila titers in only 6% of sera from patients with suspected legionellosis. Legionella-specific titers were the result of immunoglobulins G, M, and A, singly or in combination.


Rapid identification of Enterobacteriaceae from blood cultures with the Micro-ID system. Micro-ID is a new test system designed to identify members of the family Enterobacteriaceae in 4 h. It consists of 15 biochemical tests on reagent-impregnated paper disks; each test is in its own compartment in a molded plastic tray. Based on the pattern of positive and negative biochemical reactions, a five-digit octal code number is calculated. A computer-generated identification manual accompanies the product, and for each octal code listed there is a numerical value that represents the unknown isolate's degree of fit to a typical organism (LFR), a second numerical value that represents its separation from other organisms (PNOR), and a verbal description of the quality of identification. Only one reagent is added to the system. Manufacturer's directions were modified in this laboratory to allow identification from a turbid blood culture bottle within 4 h. Based on 330 routine clinical cultures tested, there was a 96.1% agreement with conventional identification to the genus and species level: 1.2% yielded first two choices possible, with one being the correct choice; 1.2% provided a correct genus, but no species identification; and 1.5% produced an incorrect identification. The Micro-ID is an accurate, facile system for the rapid identification of Enterobacteriaceae from blood cultures.


Evaluation and comparison of two assays for detection of immunity to rubella infection. Two commercially available rapid screening tests, Rubacell (Abbott Laboratories; passive hemagglutination) and FIAX (International Diagnostic Technology; indirect immunofluorescence) were compared with a standard hemagglutination inhibition assay for detection of immunity to rubella infection. In tests of approximately 300 sera, both rapid assays were specific and sensitive and showed a high predictive value of a positive result. Within-run reproducibility studies were excellent for both tests; however, Rubacell was superior to FIAX with respect to time-cost analysis.


Comparison of a semiautomated fluorescent immunoassay system and indirect immunofluorescence for detection of antinuclear antibodies in human serum. A semiautomated fluorescent immunoassay system (FIAX) for detecting anti-nuclear antibodies (ANAs) in human serum was compared with conventional indirect immunofluorescence using microscope slides (Meloy Laboratories) coated with mouse fibroblasts. The FIAX system uses quantitative indirect immunofluorescence to measure the specific binding of ANA to a sampler coated with human epithelial cells. A total of 101 serum samples were examined for the presence of ANAs by employing both methods. At an initial 1:10 screening dilution, 23 samples were negative for ANAs by the slide method, whereas 21 samples were negative with the FIAX system. Using 2+, 3+, and 4+ subjective brightness, 68 (100%) samples were positive by the slide method, whereas 67 (98.5%) were positive with the FIAX system. ANA-positive samples were diluted twofold from 1:10 to 1:640, and positive titers were determined by both methods. Sixty (77%) of 78 positive samples titrated by FIAX came within +/-1 dilution of the titers determined by the slide method, and 75 (96%) of the samples fell within +/-2 dilutions. Results indicate good correlation between the FIAX system and indirect immunofluorescence for the detection of ANAs in human serum. The FIAX system has the advantage of speed, reproducibility, and the elimination of subjective microscopic assessment of ANA titers.


Occurrence of protein A in Staphylococcus aureus and closely related Staphylococcus species. All but 1 of 143 strains of Staphylococcus aureus were positive for protein A, whereas all 34 strains of Staphylococcus hyicus and 123 of 127 strains of Staphylococcus intermedius were devoid of this cell wall component.


[Study on remodelling of bone grafts--an application of nuclear medicine (author's transl)]. The remodelling of bone grafts depends to a large extent upon the type of graft and the condition of the recipient site. We applied 99mTc-phosphate scintigraphy in a follow-up study on cases treated by bone grafting, and quantitative analysis of the scintigram by computer to make clear the difference of remodelling time or the process of acceptance between a cancellous and a cortical bone grafting or due to various graft and conditions. The result revealed that the grafted bones which were smoothly adapted for subsequent growths or functions could restore normal accumulation ratios by 36 months after the grafting. When the cancellous bone was grafted to grafting beds with good conditions such as osteotomy site in the cases with coxarthrosis deformans or congenital dislocation of the hip, it could attain the quickest recovery of the normal accumulation ratio. The next quickest recovery of accumulation ratio was attained by grafting the cancellous bone to the grafting beds with poor conditions such as osteomyelitis, pseudoarthrosis, and bone tumor. The third best accumulation ratio could be attained when the cortical bone was grafted to grafting beds with good conditions, while the slowest recovery to normal accumulation ratio was noted when the cortical bone was grafted to grafting beds with poor conditions.


Eimeria clethrionomysis sp. n., Eimeria gallatii sp. n., Eimeria pileata sp. n. and Eimeria marconii sp. n. from the red-backed vole Clethrionomys gapperi Vigors, from Pennsylvania. Four new eimerian species are described from red-backed voles, Clethrionomys gapperi in Pennsylvania. Sporulated oocysts of Eimeria clethrionomyis sp. n. are ellipsoidal, 18.8 (16.5-21.5) x 14.9 (14.0-16.5) with elongate, ovoid sporocysts, 10.6 (9.5-12.0) x6.1 (5.5-7.0). The oocyst wall is smooth, with 2 layers, and thins, with terminal cap at one or both ends. Polar granules, dark Stieda body and  sporocyst residuum are present. The oocyst residuum is absent. Sporulated oocysts of Eimeria gallatii sp. n. are ellipsoidal, 27.7 (21-32) x 19.3 (17-24) with ovoid sporocysts, 13.5 (12-15) x 8.8 (8-10). The oocyst wall is smooth, 2-layered, with a micropyle and thin wall at the end opposite the micropyle. Polar granules, Stieda body and sporocyst residuum are present. The oocyst residuum is atypical, of cobwebby material. Sporulated oocysts of Eimeria pileata sp. n. are subspherical to spherical, 25.2 (20.5-29.5) x 22.5 (19.5-25.5) with ellipsoidal sporocysts, 13.4(10.5-15.0) x 8.4 (7.5-9.5). The oocyst wall is rough, pitted, striated, 2-layered, with no micropyle. Polar granules, oocyst and sporocyst residuum, Stieda body and stiedal cap are present. Sporulated oocysts of Eimeria marconii sp. n. are ellipsoidal, 13.0 (10.5-15-0) x 10.6 (9.5-12.0) with elongate, ovoid sporocysts, 7.7 (7.0-8.5) x 4.2 (3.0-4.5). The oocyst wall is smooth, single-layered, with no micropyle. Polar granules, dark Stiedal body and sporocyst residuum are present. There is no oocyst residuum.


Trypanosoma cruzi: interaction with vertebrate cells. DNA synthesis and growth of intracellular amastigotes and their relationship to host cell DNA synthesis and growth. DNA synthesis of intracellular Trypanosoma cruzi amastigotes, following the infection of bovine embryo skeletal muscle (BESM) cells, was studied by autoradiography. After penetration, there was a prereplicative lag period (similar to or approximately 12 h) followed by a synchronous round of DNA synthesis which was found to be independent of parasite number/BESM cell cand the host cell DNA synthesis cycle. Parasite reproduction occurred, for the first time, at approximately 21 h postinfection. It was concluded that T. cruzi trypomastigotes are in the G1/G0 phase of their cell division cycle and that after penetration parasite reproduction occurs independent of events controlling host cell DNA synthesis and growth. The early synchronous growth of intracellular amastigotes should facilitate further studies on the biochemical events controlling trypomastigote-to-amastigote transformation and amastigote reproduction. A further application is envisaged for studies on the mode of action of drugs with trypanocidal activity.


Ten years' experience in general practice of dip-slide urine culture in children under five years old. Of 567 children under five registered with an industrial general practice over a 10-year period, 559 presented with symptoms of ill health in the first five years of life, and of these, 158 (27.9 per cent of those registered) had urine cultured using the dip-slide method. Thirty-four (12.3 per cent) girls and 23 (7.9 per cent) boys had at least one episode of significant bacteriuria. Two boys and three girls were found to have radiological abnormalities of the genito-urinary tract, of which two were obstructive lesions requiring surgery.Symptoms usually ascribed to the urinary tract in older children and adults did not discriminate for infection in this age group and were not a reliable indicator of the presence, or of the absence, of significant bacteriuria.The incidence of significant bacteriuria was considerably above that recorded by surveys on asymptomatic children. Proteus infections were four times more common among boys than girls, and under the age of three the proportion of boys with bacteriuria exceeded that of girls.Dip-slide culture is a valuable tool in the diagnosis, management, and follow-up of urinary tract infection.


Lack of microbial genetic response to 2.45-GHz CW and 8.5- to 9.6-GHz pulsed microwaves. Strain D4 of the yeast Saccharomyces cerevisiae, and strains TA-1535, TA-100 and TA-98 of the bacterium Salmonella typhimurium, were exposed to 2.45-GHz continuous wave or 8.5- to 9.6-GHz pulsed electromagnetic radiation (EMR) at various power densities from 1 to 45 mW/cm2. The temperature during radiation was maintained at 30 degrees C for yeast cultures and at 37 degrees C for bacterial cultures. The studies revealed no increase in mutations or of mitotic gene conversions when cells were radiated for two hours or less. Decreased viability of cells was noted in all cultures tested after radiation at power densities of 30 mW/cm2 or more; however, no reliable changes in genetic events occurred.


Determination of a thermal equivalent of millimeter microwaves in living cells. Recent microwave experiments have shown frequency dependent influences on the growth rate of bacteria. To determine whether microwaves are able to affect growth (or to induce lesions in cellular DNA of yeast cells), experiments were performed with millimeter microwaves at frequencies between 70 and 75 GHz. Saccharomyces cerevisiae cells were irradiated on millipore filter discs placed on agar plates in open petri dishes. A diploid strain of yeast (D5, Zimmerman), that is sensitive to genetic insult was used to study the effects of temperature and of microwave irradiation on cell survival, induction of mitotic recombination, and induction of cytoplasmic "petite" mutations. No evidence of altered survival, impaired function, or structural injury was seen at either frequency, even at power densities as high as 60 mW/cm2. Conventional heating had no deleterious effects until temperatures of specimens exceeded 50 degrees C. In addition, two haploid strains of yeast of opposite mating type were compared with respect to temperature and microwave treatment for formation of zygotes. The elevation of temperature due to the microwave treatment at 60 mW/cm2 and 2 mm distance was estimated to correspond to 3 degrees C.


A review of selected biological effects and dosimetric data useful for development of radiofrequency safety standards for human exposure. This report examines the bases for developing radiofrequency exposure standards which can be related to the thermogenic properties of electromagnetic fields. A review of selected biological effects, including dosimetric data and simulation of human thermodyanmic characteristics that are pertinent to standards development, is presented. Based on the analogy of thermal-stress standards that have been developed for hot industrial environments, limits on increases of body temperature are proposed as criteria for limiting exposure to radiofrequency fields, i.e., occupational exposures involving deep heating of the whole body should not increase core temperature in excess of 1 degree C. Since energy deposition from exposure to some RF fields is likely to be non-uniform and may be high in tissues that are not adapted to high rates of absorption or dissipation of thermalizing energy, means are needed to adjust focal thermal loading against the whole-body averages. A limit on core temperature is inadequate when focal elevations of temperature are close to the limits for protein denaturation, as may well occur even though the core temperature may rise less than 1 degree C. Safety limits for the general population are also discussed and here the permissible thermal load should be low enough to cause no more than an insignificant increase in core temperature. Areas needing further research to reduce the uncertainties in developing safe exposure limits for man are delineated. Even in highly adverse environmental conditions the gross thermal load and consequential heat stress from exposure to radiofrequency fields at the 10 mW/cm2 level will be small compared with that generated by any physical effort. On the basis of available data, it is concluded that the safe value for continuous exposure to 10 mW/cm2, widely used in Western countries, appears to provide an adequate margin of safety for both occupational and environmental exposure for frequencies above about 1 GHz. This limit may well be too high (perhaps by an order of magnitude) for some frequencies below 1 GHz where body resonances cause a significant increase in energy deposition and where local temperature rises occur. At the same time the present averaging period of 0.1 h seems unjustifiably short.


Ultrastructural changes in the alveolar epithelium in response to mycobacterial infection. Electron microscopic examination of the lung of mice infected with a virulent strain of M. bovis (Ravenel) revealed marked alterations in the alveolar epithelial cells, particularly Type 2 cells (granular pneumocytes), in addition to the development of interstitial and intra-alveolar granuloma. Unlike the feature in uninfected mice, more than one Type 2 epithelial cells were often found adjacent to one another within a single alveolus. Some of these cells showed mitotic figures. Their characteristic lamellar inclusions were morphologically altered.


Ultrastructural study of mycobacteria in experimentally produced lung lesions of mice. Mice were infected by iv injection with a virulent strain of Mycobacterium bovis, Ravenel strain, to prepare specimens for electron microscopical observation of their intracellular morphology. Observation was made with ultra-thin sections of the granulomatous lungs at an advanced stage of infection. Many apparently intact bacterial cells were found intracellulary, and the majority of them had lipoidal inclusions enclosed by a membranous structure. Several layers of mycobacterial cell wall were discernible, including a fairly wide space of the electron-transparent zone just beneath the electrondense outmost layer. Mesosomes, nuclear material, small dense granules and cross wall were found in almost the same appearance as those reported of mycobacteria grown in vitro. The bacilli were located mainly within intact or damaged phagosomes which were often filled with amorphous material of various electron densities.


Mechanical suture methods in esophago-gastrointestinal anastomosis. Mechanical suturing procedures have been performed in 10 patients for total gastrectomy, 6 for proximal partial gastrectomy and 7 for esophageal transection. The instrument (SPTU all-round type made in Russia) proved simple to use and produced reliable good quality closures without any other inforcement. There was one minor suture leak in 10 patients with intramediastinal esophagojejunostomy and in 6 with proximal partial gastrectomy respectively, but neither leak nor other problem has been arisen in esophageal transection for varices. It is believed that this staple suturing methods used SPTU all-round type apparatus are safe from anastomotic failure and useful for secure intramediastinal anastomosis in high level, and they represent a significant advance in the technique of thoraco-abdominal surgery.


An alternative one layer inverting suture technique for intestinal anastomosis. An alternative method of anastomosis using a suture one layer inverting was proposed in which the sutures are placed the serosa, proper muscle layer, submucosal layer and muscularis mucosae, but not mucosa. An experimental study using canine intestine revealed that tensile strength as well as bursting pressure at the anastomotic site is significantly superior in our method to the end-on method.


Retroperitoneal hemangiopericytoma associated with hypoglycemia: report of a case. A 34-year-old woman had episodes of hypoglycemic attack 8 years after a surgical resection of a retroperitoneal hemangiopericytoma. In spite of normal levels of serum IRI, insulin radioreceptor assay demonstrated high level of plasma ILA (insulin like activity). The patient underwent resection of a recurrent retroperitoneal tumor with metastatic lesions of the liver. Postoperatively, ILA level in plasma by insulin radioreceptor assay decreased, and hypoglycemic attacks disappeared. Therefore, this associated hypoglycemia was presumed to be not caused by excess glucose consumption by the tumor, not by excess secretion of IRI by the tumor, but caused by the presence of high level of ILA related to the tumor.


Growth control of normal and transformed cells. Both serum factors and protein synthesis are required for normal cell growth. Swiss 3T3 cells require the serum growth factors insulin and EGF (epidermal growth factor) during the initial part of the G1 period, until they pass a restriction point about 2 h before the initiation of DNA synthesis. Concentrations of cycloheximide that inhibit protein synthesis by as much as 70% dramatically lengthen the cell cycle before the restriction point, while the cell cycle after the restriction point remains nearly constant. These results are consistent with a model in which labile proteins are required for transit of cells past the serum-sensitive restriction point. The relation of these findings to the growth control of transformed cells is discussed.


Possible role of fibronectin in malignancy. Frozen sections of tumors induced by injecting virally transformed cells into animals were stained for fibronectin by immunofluorescence. Many tumor cell lines do not express fibronectin in tumors in situ even though some of them express fibronectin in culture. Cell shape and hormones appear to influence the expression of fibronectin in culture; however, it is unclear how fibronectin expression is regulated in vivo.


Evidence of microfilament-associated mitochondrial movement. The mitochondria in the lower Malpighian tubule of the insect Rhodnius prolixus can be stimulated by feeding in vivo and by 5-hydroxytryptamine in vitro, to move from a position below the cell cortex to one inside the apical microvilli. During and following their movement into the microvilli, the mitochondria are intimately associated with the microfilaments of the cell cortex and microvillar core bundle. Bridges approximately 14 nm in length and 4 nm in diameter are observed connecting the microvillar microfilaments to the outer mitochondrial membrane and microvillar plasma membrane. Depolymerization of all visible microtubules with colchicine does not inhibit 5-HT-stimulated mitochondrial movement. On the other hand, treatment with cytochalasin B does block mitochondrial movement, suggesting that microfilaments play a role in the mitochondrial motility. We have labeled the microvillar microfilaments, which are 6 nm in diameter, with heavy meromyosin, which supports the contention that they contain actin. A model of the mechanism of mitochondrial movement is presented in which mitochondria slide into position in the microvilli along actin-containing microfilaments in a manner analogous to the sliding actin-myosin model of skeletal muscle.


Action of phorbol esters in cell culture: mimicry of transformation, altered differentiation, and effects on cell membranes. The carcinogenic process is usually multifactor in its causation and multistep in its evolution. It is likely that entirely different molecular mechanisms underlie the many steps in this process. In contrast to initiating carcinogens, the action of the tumor-promoting phorbol esters does not appear to involve covalent binding to cellular DNA and they are not mutagenic. Recent studies in cell culture have revealed two interesting biologic effects of the phorbol esters and related macrocyclic plant diterpenes. The first is that at nanomolar concentrations they induce several changes that resemble those seen in cells transformed by chemical carcinogens or tumor viruses. These include altered morphology and increased saturation density, altered cell surface fucose-glycopeptides, decrease in the LETS protein, increased transport of deoxyglucose, and increased levels of plasminogen activator and ornithine decarboxylase. In transformed cells exposed to phorbol esters the expression of these features is further accentuated. Phorbol esters do not induce normal cells to grow in agar but they do enhance the growth in agar of certain transformed cells. The second effect of the phorbol esters is inhibition of terminal differentiation. This effect extends to a variety of programs of differentiation and is reversible when the agent is removed. With certain cell culture systems induction of differentiation, rather than inhibition, is observed. Both the transformation mimetic and the differentiation effects are exerted by plant diterpenes that have tumor-promoting activity but not by congeners that lack such activity. The primary target of phorbol esters appears to be the cell membrane. Early membrane-related effects include enhanced uptake of 2-deoxyglucose and other nutrients, altered cell adhesion, induction of arachidonic acid release and prostaglandin synthesis, inhibition of the binding of epidermal growth factor to cell surface receptors, altered lipid metabolism, and modifications in the activities of other cell surface receptors. A model of "two stage" carcinogenesis encompassing the known molecular and cellular effects of initiating carcinogens and tumor promoters is presented. According to this model, initiating carcinogens induce stable alterations in the cellular genome but these are not manifested until tumor promoters modulate programs of gene expression and induce the clonal outgrowth of the initiated cell.


Jaborandi: an interdisciplinary appraisal. In spite of many references to Pilocarpus Jaborandi Holmes in ethnological and botanical sources and suggestions of its employment for a variety of diseases, it has not been possible to pin down the use of its leaves to any particular purpose amongst South American Indians. While the medically important jaborandis are species of Pilocarpus, it is true that this vernacular name is commonly applied to other rutaceous and numerous piperaceous plants as well. The introduction of jaborandi leaves to western medicine goes back to 1873, when Symphronio Coutinho went to Europe, taking with him samples of the leaves. The copious sweating and salivation brought about by the leaves attracted the attention of French physicians. Soon jaborandi leaves were being employed in the treatment of many diseases. In 1875, Hardy and Gerrard independently discovered the alkaloid pilocarpine. Most therapeutic applications of jaborandi leaves and pilocarpine fell into disuse and were discontinued. What remained was the use of the latter in ophthalmology, where it had been introduced as a miotic by Weber in 1876. The mixture of pilocarpine and another natural product, physostigmine, remains to this day one of the mainstays in ophthalmology.


Chemical studies on medicinal Myristicaceae from Amazonia. Drugs from Myristicaceae species are used in the Amazon region as hallucinogens and arrow poisons, as well as for the healing of infected wounds. The former effects were attributed by Schultes and Holmstedt to tryptamines and carbolines. The latter activity is now tentatively ascribed to pterocarpans and neolignans. This, and a proposal that the red colour of the bark resins may be due to oxidative dimers of flavans, are based on a review of the chemistry of Amazonian Myristicaceae.


Arrow poisons in China. Part I.

Arrow poisons in China. Part I. Arrow poisons have been used for at least 2500 years in various parts of China by the Han and other peoples. The preparation and use of these poisons is discussed on the basis of accounts in Chinese and Western sources. Mostly, the principal ingredient has been an extract derived from the tubers of Aconitum species, especially A. carmichaelii Debx. (wu t'ou, fu tzu, ts'ao wu). Certain peoples of the south-west and south, besides using Aconitum, have also obtained an essential ingredient from the juice (latex) of Antiaris toxicaria Lesch. (tu mu, hu, chien hsüeh feng hou, nu chien tzu, ka tuk). The chemistry and pharmacology of the active principles found in certain of the plants incorporated into the poisons are dealt with briefly (but this does not include Aconitum, which will be treated in some detail in Part II).


Stereotaxic electrode placement in the neonatal rat. A head holder for stereotaxic electrode placement in the neonatal rat is described. The neonatal head is held in a fixed and reproducible position by means of a mouth bar and a recurved needle hooked into the foramen magnum. With the aid of this instrument, a stereotaxic atlas for the 3-day-old rat has been prepared. The head holder has been used for rats up to 10 days of age and permits electrode placement in the neonatal rat in a manner essentially analogous in ease and precision to that of stereotaxic methods for the adult animal.


A double constriction micropipette for the quantitative injection of tracer substance. A glass, double constriction injector micropipette ('micropet') has been designed to overcome problems associated with commercial microliter syringes. These problems include: inconsistent volume delivery, inability to view tracer during filling or injection, and the relatively large size of the needle supplied. 'Micropets' of different volumes are easily made from inexpensive, commercially available Drummond 100 microliter glass tubes (bores) fitted with teflon plungers. Tracer substances may be viewed in the bores during filling and injection and the needle tip can be pulled as small as 50 micrometers. For deep stereotactic injection, a long triple constriction 'micropet' has also been devised. The 'micropet' is based in principle on the commercial Lang-Levy double constriction micropipettes and these are used for radiometric calibration of the 'micropets'. Thus the 'micropet' accuracy is a reflection of these commercial micropipettes, and is approximately +/- 0.5%. Precision is also on the order of +/- 0.5% as determined by radioassay. When combined with an oil-filled, variable-speed microdrive the injector 'micropet' provides a highly controlled and precise system for the accurate delivery of tracer substance important for quantitative neuroanatomical investigations.


A method for recording single unit activity from the brains of foetal pigs in utero. A technique is described for recording single unit activity from foetal miniature pigs in utero. The method involved anaesthetizing the sows with halothane and exteriorizing a sufficient length of the uterus to contain a single foetus. Both foetus and uterus were placed in a stereotaxic frame which was held independent of the mothers' support. Preliminary analysis of recordings obtained from spontaneously firing single nerve cells in the post-central cerebral cortex indicate that the patterns of spike activity in prenatal pigs are very similar to those observed in prepubertal animals.


A simple and rapid section embedding technique for sequential light and electron microscopic examination of individually stained central neurons. A method for section embedding of central nervous tissue, and its application to light and electron microscopic examination of neurons stained by intracellular injection of horseradish peroxidase, is described. After primary fixation in aldehydes, thick (10--80 micrometer) sections are cut on a Vibratome. They are then treated for the histochemical demonstration of peroxidase, postfixed in osmium tetroxide, dehydrated and infiltrated with Spurr's low viscosity resin. Infiltrated sections are embedded between glass slides and coverslips coated with a transparent layer of teflon and polymerized. The resulting mount allows protracted storage and high resolution light microscopic examination of sections. When desired, the glass components can be removed and the specimen cemented to a blank block for thin sectioning and electron microscopic examination.


Variations in the cell cycle of slow-growing Escherichia coli 15T-. DNA synthesis was measured via 3H-thymidine pulses, and patterns of incorporation were compared to cell number increases during synchronous division of Escherichia coli 15T- (555-7) in M9.01% aspartate. The timing of chromosome initiation, duration of chromosome synthesis, and the D period were found to vary over the three cycles of division analysed. In addition, the D period measured by comparing 3H-thymidine incorporation and concomitant division, differed from D period measurements made by timing division cessation in DNA synthesis inhibitors.


Microcalorimetry studies of energy changes during the growth of Klebsiella aerogenes in simple salts/glucose media. 1 Establishment of standard conditions. The shape of the thermogram and the total heat output of aerobically growing cultures of Klebsiella aerogenes depend on the nature and state of the inoculum, the composition of the growth medium, aeration in the fermentor and in the calorimeter, and the pump rate of the culture through the microcalorimeter cell. Standard conditions of growth and heat measurement have been established which give reproducible thermograms and total heat output. Experimental results using different glucose concentrations are reported and the enthalpy changes for the consumption of glucose and the production of cells have been calculated.


[Prophylactic treatment of pancreatitis with antibiotics? (author's transl)]. In acute pancreatitis the standard treatment is with antibiotics given prophylactically. Available studies indicate that the usefulness of such chemotherapy in the milder form of the disease (degree of severity I) has not yet been fully established. In the absence of contradictory results, prophylactic antibiotic treatment can, however, be recommended in severe pancreatitis (degrees II and III) and in those conditions induced by ethyl or of idiopathic origin. In all forms showing underlying biliary tract disease and in pancreatitis complicated by infection, antibiotic treatment should invariably be administered. Administration of broad-spectrum antibiotics which can be excreted via the bile duct, e.g. ampicillin or tetracycline, is preferable.


Complement activation in pneumonia.

Complement activation in pneumonia. Complement analyses performed on serially collected plasma samples from 10 patients during acute infectious pneumonia or bronchopneumonia showed normal or increased values for hemolytic activity and Clq, C4, C3, and factor B levels. However, the levels of C3 breakdown products (C3d) were significantly (greater than 2 SD) increased in six patients during the first three days of observation, suggesting that hypercatabolism of complement components may occur during the acute phase of infectious pneumonia concomitant with a hypersynthesis of some complement components. Evidence of circulating immune complexes was obtained only in two patients with the 125I-Clq Binding Test.


The autosomal recessive (Becker) form of myotonia congenita. In the last two decades, two genetically distinct forms of myotonia congenita have been identified--an autosomal dominant and an autosomal recessive form. The purpose of this review is to describe the features that enable us to distinguish between these two forms in the absence of sufficient genetic data. Thus far, it can be concluded that the only probable difference between the two forms is in the fatty-acid pattern of muscle phospholipids. Clinical, histologic, ultrastructural, and electromyographic investigation may prove helpful, but they alone cannot provide a reliable means of identifying the genotype in an individual patient.


Lack of benefit of allopurinol in Duchenne dystrophy. A double-bline, controlled study was conducted to assess the effects of allopurinol on Duchenne muscular dystrophy. Six patients--three treated with allopurinol and three given a placebo--were followed up for one year and were evaluated by functional and manual muscle testing as well as by determination of serum creatine kinase (CK) levels. Patients in both the allopurinol-treated group and the placebo group deteriorated at varying rates without evidence of therapeutic benefit. To exclude the possibility of error attributable to small sample size, all six patients were treated with allopurinol for an additional eight-week period. Muscle strength continued to deteriorate during this phase of the study.


Congenital hypotonia revisited.

Congenital hypotonia revisited. The definition of neuromuscular diseases affecting infants has depended on factors as various as the rate of progression of the illness, the clinical picture, and, recently, the morphologic peculiarities in the muscle biopsy. A review of the literature suggests that there are discrepancies in the classification of such illnesses, no matter what system is used. In some instances, a single diagnosis seems to include patients with quite separate illnesses, whereas other patients with seemingly identical diseases have been given different diagnoses.


Physiologic vs pathologic myopia: genetics vs environment. Physiologic myopia occurs as the result of a correlation failure of refraction components in the normal eye. Pathologic myopia is caused by excessive axial elongation that primarily involves the ora-equatorial area and the posterior pole. Peripheral fundus changes and posterior staphyloma formation are ophthalmoscopic evidences of this process. Heredity is the basic determinant of ocular refraction, but numerous agents produce both temporary and permanent myopias.


ELISA for diagnosis of ocular toxocariasis. The enzyme-linked immunosorbed assay test was positive in 37 of 41 patients (90%) suspected of having clinical Toxocara infections. The antigen used in this test is prepared from the Toxocara egg and is felt to be extremely specific in making the diagnosis. The prevalence of Toxocara infection is much greater than previously believed. It is hoped that this test will help the surgeon fell more secure in not enucleating eyes containing masses suggestive of Toxocara infection.


Indications and surgical techniques for orbital exenteration. Indications for the mutilating operation of exenteration are enumerated. They usually involve a malignant neoplasm of the orbital contents, primary, direct extension, or adnexal tissue that cannot be controlled by simple excision or irradiation. Surgically, subtotal exenteration with partial preservation of lids and even conjunctiva may be achieved occasionally. However, total exenteration may be lifesaving. Techniques and precautions are discussed. Advantages and disadvantages of skin grafting that influence the postoperative care are noted.


[Carcinomas of the digestive tract. Chemotherapy as a complement to surgery (author's transl)]. Chemotherapy as a complement to surgery or adjuvent chemotherapy has been presented as one of the possible means of improving 5 years survival rates following the excision of a digestive tract carcinoma. Started early after excision, and given in adequate doses for a period of one to two years, it is aimed at treatment of residual sub-clinical disease. Its effectiveness may be assessed only by study of the reduction in the number of recurrences or metastases in a large number of patients treated. At the present time, published series have not made it possible to define whether there is a definite reduction in the percentage of recurrence and metastases or merely a prolongation of the clear period which precedes their clinical manifestation. The practical difficulties of treatment, together with its possible early or late toxicity, are such that it should be reserved for forms in which there is some doubt as to the prognosis. The addition of immunotherapy has to be defined.


[Hyperselective blood sampling from the renal veins (author's transl)]. A method by substitution of venous catheters within the renal venous tree was used in 28 adult hypertensive patients suffering from focal parenchymatous lesions or stenosis of one branch of the renal artery with the aim of detecting a specific source of renin hypersecretion.