Studies in vitro on the effect of adenosine, adenine and NAD on guinea pig myometrium in the presence of exogenously applied PGE2.

In guinea pig isolated uterine strips, separate addition of adenosine, adenine and NAD was without any effect. Adenosine and NAD when administered after previously applied subthreshold dose of PGE2 exhibited a strong contractile effect. Indomethacin, added to the organ bath before addition of prostaglandin, blocked, partially or totally, the observed synergistic action of PGE2 with adenosine and NAD. From the experimental data of the present work it is postulated that adenosine and NAD stimulate prostaglandin biosynthesis in myometrium as already it has been suggested for ATP, ADP and AMP. The aspect for the existence of a functional link between adenyl compounds and prostaglandins, based on these findings, is reinforced.


Effects of 2-deoxy-D-glucose on the contraction frequency of the isolated atria from fed and fasted rats.

The mere exogenous substrate removal did not change the contraction frequency of the isolated rat atria. However, addition of 2-deoxy-D-glucose together with the glucose removal, elicited a decrease in the atrial frequency. This decrease was significantly greater in the atria from fed rats with respect to those from fasted rats. Near the end of the experiments, only in the atria from fed rats, transient irregular beating appeared. The results suggest that triglycerides constitute the major endogenous substrate of the sinus node pacemaker cells when rats have been previously fasted and that these cells have metabolic features similar to those of contractile fibres.


Glucose metabolism by isolated fat cells from sloth.

Glucose metabolism by sloth fat cells with and without addition of insulin was investigated. The data were compared to the results obtained with rat fat cells incubated under the same experimental conditions. Sloth fat cells showed a very low glucose oxidation to 14CO2 and incorporation into total lipids. The glucose incorporated into lipids is mainly in the glyceride-glycerol moiety. Addition of insulin did not produce an increase on glucose oxidation and a slight increase in the incorporation into total lipids was observed. Since it has been reported that sloths have a very low rate on thyroxine secretion, the results are discussed in relation to data in the literature on carbohydrate and lipid metabolism in hypothyroid animals.


The rat and hamster jejunum during transintestinal transport in vitro.

By using the jejunal tract of rat and hamster intestine, net fluid transintestinal transport in the everted sac incubated in vitro at 28 degrees C and at 38 degrees C has been determined. In the scraped mucosa wet weight/dry weight ratio, cell water, sodium and potassium concentration have been detected in vivo and in vitro, throughout the incubation time, at the two different temperatures. Under all these conditions ATP, ADP and AMP levels have been determined in total intestine and in scraped mucosa. In rat jejunum incubated in vitro at 38 degrees C transintestinal fluid transport continuously decreases during 1-h experiment; in the same time the enterocyte gains sodium, dilutes cell potassium and swells, whereas its energy charge is dramatically reduced. All these parameters are constant in rat jejunum incubated at 28 degrees C and in the hamster incubated both at 28 degrees C and at 38 degrees C throughout the experiment. An inadequate diffusion of oxygen into the enterocyte, could tentatively explain experimental results obtained on rat jejunum at 38 degrees C. Under all tested conditions, energy charge and intracellular potassium are lower in conditions in vitro than in those in vivo; the contrary happens for cell sodium and swelling. The oxygenation of the intestine in vitro, lower than the one in vivo, could explain the different behaviour found in the two experimental conditions.


The effect of training and detraining on several enzymes in horse skeletal muscle.

Training and detraining had little effect on the activity of glycogen synthase, hexokinase, glycerol 3-phosphate dehydrogenase or total protein. The activity of 3-hydroxyacyl-CoA dehydrogenase increased markedly during training. After 5 weeks of detraining, the activity of 3-hydroxyacyl-CoA dehydrogenase was returning to pre-training values, whilst by 10-week detraining, the levels were increasing again.


[Effect of pindolol on tachycardia and certain metabolic variations dependent of muscular exertion].

1. -- In young well trained adults, adrenergic beta receptors blockade by pindolol or propranolol causes a relative bradycardia during running on a threadmill during 10 to 20 min at 50% VO2 max. 2. -- Pindolol activity is maximum for 5-7.5 mg ingested one hour before starting. In two subjects, tachycardia reappears for 30 mg : this secondary increase may be explained by an intrinsic sympathetic activity of pindolol. 3. -- Propranolol is about 14 times less active than pindolol without any intrinsic sympathetic activity.


[Experimental use of prostaglandin F2 alpha to synchronize estrus in cattle].

Experiments were conducted for the use of prostaglandin to synchronise oestrus in dairy cows. One single dose of 5 mg F2-Alpha prostaglandin was administered via the intrauterine route. The reference values of the control groups were matched by first insemination following induced oestrus among the best of the test animals which were inseminated twice. These values reached 72 and 96 hours following application accounted for 46.7 and 50.0 per cent. The yellow body phase, a prerequisite for oestrus production by prostaglandins, had been induced in the test animals by cycle blocking, using gestagen. Possible use of the method in a bio-engineering context for large-scale oestrus synchronisation in cows is discussed.


Human adenohypophyseal quantitative histochemical cell classification. I. Morphologic criteria and cell type distribution.

Recent advances in immunocytochemistry and histochemical staining have made it possible to identify pituitary cells that contain specific hormones by their appearance under light microscopy. Criteria for identification of functional cell classes were formulated from these studies. Fourteen hypophyses were obtained at the time of autopsy (four to 22 hours postmortem), embedded in paraffin, sectioned, and differentially stained. Cell counts on areas in the acidophil wing and basophil wedge were performed using the formulated criteria. Chromophilic thyrotrophic hormone cells and melanocorticotrophic hormone cells occurred significantly more frequently in glands from men, whereas the numbers of other cell types were statistically comparable in glands from both sexes. We were able to functionally classify 72.1% of 53,167 pituitary cells. Only 0.1% of presumptive secretory cells were completely chromophobic. Cell distribution patterns were similar to those previously described, with the exceptions that gonadotrophic cells were more prominent in the lateral wings and that prolactin cells had a more even distribution.


Special problems and indications of the palliative osteosynthesis of the cervical vertebra.

The palliative internal fixation of the cervical spine becomes more and more important. Independent of the patients age, the procedure is indicated by neurologic complications, very severe pain, in patients of generally good condition. The tumorous lesion of the spine should be a local restricted one.


Resolution of block neutralization test curves into components of the foot-and-mouth disease virus system.

Classic neutralization studies by Fazekas de St. Groth and Webster (8) on mixtures of influenza viruses and mixtures of rabbit antisera are reinterpreted in terms of a percentage contaminant in the stock used for the dilution series. A very small amount of a different virus changes the shape of quantal assay curves considerably, but even a large amount of a different antiserum has negligible effect on the shape and merely shifts the curve along the serum dilution axis. These conclusions are the reverse of the authors, who only considered an absolute amount of another component in all dilution tubes. An artificial mixture of 2.7 percent O8 in O1 foot-and-mouth disease virus strains was tested against anti-O1 serum assaying in suckling mice. The small amount of O8 virus greatly altered the shape of the neutralization curve in the direction expected from the reanalysis of the influenza literature data. Results from artificial mixtures are used to explain what were hitherto anomalously broad neutralization curves for some other foot-and-mouth disease strains given by Booth et al. (1) Many of the virus stocks studied can now be postulated as a natural mixture of related virus strains. In fact, the O1 and O8 stocks used might also be of themselves mixtures. These virus strains also exhibit still a further test complication in that the virus-antibody reaction appeared to shift away from complexes on dilution immediately prior to assay.


[Analysis of the seasonal pattern of blood lead levels in relation to different concentrations of atmospheric lead].

Blood lead levels have been analysed in a group of people resident in Turin in relation to different concentrations of lead in the atmosphere. Blood samples were taken in three periods of the year corresponding to seasonal maximums of pollution. Blood lead levels showed a marked decrease in the summer when the lowest concentrations are observed but the following autum sample did not show an increase. It is therefore considered likely that blood values would have been found to be increased if the final sample had been put off by 1-2 months.


Tracing neural connections of human brain with selective silver impregnation. Observations on geniculocalcarine, spinothalamic, and entorhinal pathways.

In selected human brains, it is possible to study the efferent connections of a damaged site with the suppressive silver impregnation techniques described by Nauta and Gygax. Autopsy specimens with circumscribed lesions of recent origin ( one to five weeks before death) are suitable. However, the large size of the human brain and the lack of perfusion with fixative necessitates modifications in the methodology used on experimental animals. With these modifications, it has been possible to trace details of the spinothalamic tract, the geniculocalcarine pathway, and a projection into the entorhinal area in autopsied human brains. More frequent use of this methodology may substantially increase the information that is currently available on the neuronal connections of the human brain.


[Carbon monoxide: environmental and biological aspects].

The problem of carbon monoxide as a pollutant is examined. Its physico-chemical features are considered along with the sources of pollution (natural and technological), the characteristics of the pollutant in the environment and, particularly, the effects of car and industrial emissions and of cigarette smoke on the human organism.


Detection of B cell antigens in multiple sclerosis. Use of serum from multiparous women.

Sera from two multiparous wives of patients with multiple sclerosis (MS) were used to detect B cell antigens in other patients. With serum X, 11 of 16 patients were positive as compared with ten of 16 controls (.05 less than P less than .1). With serum Y, a positive response was found in 11 of 16 patients and two of 23 controls (P less than .0005). Ten of the 11 patients who were positive with serum Y were also HLA-Dw2, which suggests that the B cell antigen detected by this serum is linked to Dw2. Three of four Dw2-positive controls were negative with serum Y, which raises two alternative hypothetical possibilities concerning the B cell antigen. These findings indicate that serum from multiparous wives may be an important tool in the investigation of the genetic components associated with MS.


Salvage operations for malignant obstruction of the oesophagus.

Two patients with locally advanced carcinoma of the oesophagus, who did not obtain palliation with radiotherapy, underwent salvage operations to restore their ability to swallow. One patient was subjected to pharyngolaryngo-oesophagectomy together with excision of the involved posterior tracheal wall. Reconstruction was by pharyngoglossogastrostomy, with a deltopectoral flap used to replace the tracheal defect. The second patient, who had cervical metastases and severe pulmonary complications, underwent a jejunal loop bypass operation. Both patients were able to eat normally before dying many months later from metastatic disease. Salvage operation should be considered for patients with primary tumours which appear to be slow-growing, even though locally advanced and free of widespread disease. Worthwhile palliation is obtained when the operation, either resection or bypass, has been successful.


Amniotic fluid beta 2 microglobulin: correlation with L/S ratios and creatinine levels.

Beta 2 microglobulin (beta 2 m) levels were measured in amniotic fluids taken after the 32nd week of gestation. Beta 2 m levels fell continuously from 5.2 microgram/ml at the 32nd week to 3.08 microgram/ml after the 39th week of gestation. Abnormally high levels of beta 2 m were found in 12.6% of samples. There was a significant correlation between beta 2 m levels and lecithin/sphingomyelin ratios at 32 weeks of gestation and with creatinine levels at 37-38 weeks (P less than .05). Although elevated beta 2 m levels may be associated with maternal and/or fetal abnormalities, the precise relationship is not clearly established: in particular, our data do not suggest that estimation of beta 2 m levels could be of value in determining fetal maturity.


Ultrastructural analysis of the lining layer of the rat pulmonary alveolus.

Using electron microscopy we have examined the lining layer of the rat pulmonary alveolus. This layer appears as a morphological entity 1-3 days after birth: it is composed at first of a filamentous Ruthenium Red-negative material derived from lamellar bodies, and subsequently (4 days after birth) of a homogeneous Ruthenium Red-positive material. This latter material, which corresponds to the epithelia lining of the alveolus typical of adult rats, is presumably derived from a mixture of the filamentous material produced by the lamellar bodies, and a material produced by the alveolar cells 4 days after birth which contains acidic groups which bind Ruthenium Red.


A case of acute myelomonocytic leukaemia associated with therapy: Feulgen-DNA microdensitometric determinations.

A cytochemical study of a case of acute myelomonocytic leukaemia, which arose after radiation therapy and polychemotherapy for Hodgkin's disease, is presented. The distribution of the DNA content of the circulating blood cells appears to be different from the unimodal diploid distribution reported in the literature for acute myelomonocytic leukamias not associated with therapy. This finding seems to strengthen the hypothesis, already put forward by Marinone et al. (1979), that a particular pattern of DNA content distribution could be characteristic of leukaemias linked to chemotherapy and radiotherapy.


Feulgen-DNA content of the Purkinje neuron: "diploid" or "tetraploid"?

Microdensitometric measurements of the Feulgen-DNA content of the Purkinje cells and of the small granular cells (2c control) were carried out at lambda 550 nm and by the two wavelength method according to Fukuda et al., and were also corrected for the glare. We analysed sections of perfused cerebella and isolated cells of cerebellar cortex from adult rats. The Purkinje cells had a mean value of Feulgen-DNA content 30-45% higher than the 2c value, irrespective of the methods of preparation or measurement.


The relation of heart rhythm to postextrasystolic potentiation.

In 9 patients with various forms of heart rhythms and rates, all of whom had a temporary pacemaker electrode inserted, several single premature ventricular contractions (PVC) with various coupling intervals (CI) were induced. The relative systolic area of the carotid pulse curve in the postextrasystolic beat was used as an indicator of the degree of postextrasystolic potentiation (PESP). A close correlation between PESP and CI was found only in sinus rhythm patients (r = -0.85). The patients with junctional and ventricular rhythms had no PESP regardless of CI or the degree of heart failure. Compensatory pause (CP) in these patients surprisingly related (r = 0.95) to heart rate, slower rates having a shorter relative CP.


Quantitative determination of cardiac myosin subunits stained with fast green in SDS-electrophoresis gels.

In the present study a procedure is described which permits simultaneous relative quantification of all myosin subunits in the same fast green stained electrophoresis gel. The relative protein quantification can be performed successively in the same gel using densitometry and the elution method, respectively. Thus, the results obtained with one method can be checked with the other. This procedure provided reliable estimates of the stoichiometry of cardiac myosin subunits within a range of about 20--110 micrograms of total myosin protein per elctrophoresis gel.


Increase in fatty acid synthetase content of 3T3-L cells undergoing spontaneous and chemically induced differentiation to adipocytes.

3T3-L fibroblasts differentiate into adipose cells when maintained in a non-growing state. The specific activity of fatty acid synthetase of differentiated cells was 25--30-fold higher than that present in 3T3-L fibroblasts or in 3T3-C2 cells that possess an extremely low incidence of differentiation to adipocytes. The results of immunochemical analysis indicate that the increased specific activity of fatty acid synthetase in the differentiated cells is due to an increase in the cellular content of this enzyme. The rate of conversion of adipose cells was accelerated by brief exposure of confluent non-growing cultures of 3T3-L cells to 3-isobutyl-1-methylxanthine. This was accompanied by an increase in the specificity activity of fatty acid synthetase, which was also shown to be due to an increase in the cellular content of this enzyme. The continuous presence of 3-isobutyl-1-methylxanthine in the culture medium was not required to elicit the morphological and biochemical changes in 3T3-L cells that occurred many days after the removal of the inducer but earlier than the onset of spontanous differentiation.


Immunologic regulation in pregnancy.

Theories to explain the failure of rejection of the fetus by the mother during pregnancy include immunologic privilege of the uterus as a graft site, lack of transplantation antigen expression on the trophoblast, weakening of maternal cellular immunity during pregnancy, and separation of maternal and fetal circulations. Evidence for and against each of these theories is discussed. Local concentration of a variety of hormones, including human chorionic gonadotropin (HCG), sex steroids, alpha-fetoprotein, and immunoglobulins, could provide a blocking mechanism to prevent maternal cellular immune attack. Possibly, progesterone, antibodies, and immune complexes are important in protecting the placenta and ultimately the fetus from rejection. Elucidation of regulatory mechanisms in pregnancy may be applicable to other problems in immunology.


Placenta-bound immunoglobulins.

Low pH eluates were prepared from trophoblasts derived from 8 term human placentas. A qualitative analysis for immunoglobulins revealed the presence of IgG, IgA, and IgM in these eluates. IgC-rich fractions were obtained by DEAE-cellulose chromatography of ammonium sulfate-concentrated eluates. These fractions were able to neutralize, in vitro, the catalytic activity of reverse transcriptases (RT) from several retroviruses. RT from baboon endogenous virus (BEV) seemed to be more susceptible to the neutralizing activity of some eluates. This was in contrast to RT from feline leukemia virus (FeLV) which were neutralized by eluates of leukocytes from chronic myelogenous leukemia. In contrast to previous and present results with purified IgG from leukemic leukocytes, the purified IgG from placenta eluates was incapable of RT neutralization. However, such purified IgG fractions inhibited mixed lymphocyte reactions.


[Modifications in alpha 2 globulins, gamma globulins and in rheumatoid factor during gold salt therapy in rheumatoid arthritis].

A total dose of g 1.071, given as hydrosoluble salts for a 12 month period, showed a significant decrease in serum gamma globulins along with clinical improvement in 17 patients affected with rheumatoid arthritis. A decrease in alpha 2 globulins and in rheumatoid factor titre was observed too, but it was not significant. The data suggest that in rheumatoid arthritis the gold therapy might also be effective on the immunological disease mechanism.


[Various histochemical aspects of chorionic villi].

Detail studies on a diastasic digested leucofuxin coloured citotrofoblastic and synciziotrophoblastic villus cells permitted to localize chorion glicoprotein concentration almost exclusively in the proximal portion of the trophoblastic syncitium. Furthermore, like in the epatic tissue, but unlike the findings in some other sites such as sarcolemma, necrilemm basal membranes, the reticula villus substance is argirofila but not P.A.S. positive. Some considerations were proposed to explain this abnormal pattern.


[Current modifications in lipoprotein staining method on acetate gel cellulose].

Some modification are proposed to standardize better the staining technique of lipoprotein fractions. These modifications facilitate greatly the procedure originally reported by Colfs and Verheyden.


Immunological cross-reactivity between acid extracts of myelin, liver and neoplastic tissues: studies in immunized guinea-pigs.

Groups of 4 guinea-pigs were immunized with acid extracts prepared from bovine myelin (EF), normal human liver tissue and malignant or benign neoplastic tissues in Freund's complete adjuvant (FCA1. The animals were weighed daily and examined for clinical signs of experimental allergic encephalomyelitis (EAE). All the animals immunized with EF developed clinical symptoms of EAE within 21 days of the initial immunization, whilst some of the animals immunized with certain tumour extracts developed symptoms which closely resembled those of EAE. Control animals immunized with FCA only remained asymptomatic. Cellular immunity to the various extracts in immunized animals was assessed 20 days after immunization by i.d. skin testing, and upon killing at Day 21 with the direct peritoneal-exudate macrophage migration inhibition (MMI) test. Brains and spinal cords were removed at killing, fixed in formalin and processed for histological examination. I.d. skin testing was shown to be most consistent in demonstrating positive delayed hypersensitivity, whilst the MMI test frequently gave negative results in the presence of pronounced skin responses to specific extracts. Thus it was shown that 3/4 animals immunized with basic proteins extracted from an adenocarcinoma of the lung or related hepatic metastases, and 1/2 animals immunized with an extract of a carcinoma of the breast, gave intense erythema and induration responses 5 mm in diameter 24 h after i.d. challenge with EF. No such response was obtained in animals immunized with basic proteins extracted from normal human liver, any of the other neoplastic tissues, or in control animals immunized with FCA only. Examination of brains and spinal cords from animals immunized with EF revealed dense infiltration by mononuclear cells in the ependyma and choroid plexus of levels in the spinal cord. Examination of brains and spinal cords from animals immunized with the lung-tumour extract or related hepatic metastases which showed demonstrable immunological cross-reactivity with EF in immunized animals, revealed a number of inflammatory changes characterized by dense infiltrates of mononuclear cells sub-ependymally, and perivascular cuffing in the cortex. However, no significant lesions were seen in the spinal cords of these animals. Polyacrylamide-gel electrophoresis of the 2 tumour extracts exerting this apparent encephalitogenic effect did not reveal proteins within the mol. wt range of EF. Thus the observed pathological effects and cross-reactivity with EF were probably not due to contamination with nervous-tissue components. It is suggested that these tumour extracts may have contained a component or components other than EF, immunologically cross-reactive with EF, and capable of inducing the observed encephalitis.


Isolation of plasma-membrane components from cultured human pancreatic cancer cells by immuno-affinity chromatography of anti-beta 2M sepharose 6MB.

Human pancreatic exocrine adenocarcinoma cells established in tissue culture expressed both HLA and beta 2-microglobulin (beta 2M). Plasma-membrane components of this pancreatic cancer cell line were purified from plasma membrane fractions enriched by sucrose density-gradient centrifugation, using immunoaffinity chromatography on immobilized anti-human beta 2M antibody. Both rabbit and mouse monoclonal anti-beta 2M IgG were used, with a 20--25-fold overall purification of 5'-nucleotidase. The method was applicable to 5 x 10(7) cells and permitted the solubilization of membranes retained on the column, with the selective desorption of components not associated with beta 2M before the subsequent elution at pH 3 of beta 2M-associated macromolecules. The acid eluate contained one major and two minor bands in the 40--45,000 mol.-wt range with two additional enriched components of 18,000 and 22,000 dalton. A major carbohydrate-containing component of high mol. wt was also found to be associated with the pancreatic cancer-cell plasma membrane.


Tumour markers in breast cancer.

The clinical usefulness of 8 potential tumour markers has been evaluated in 69 patients with Stage I and II breast cancer and 57 patients with Stage III and IV. Serum CEA concentrations were raised in 13% of patients with local and 65% of those with advanced breast cancer. In patients with clinical evidence of progression or regression of tumour, serum CEA levels changed appropriately in 83% of cases. Taking 4 of the markers (carcinoembryonic antigen (CEA), lactalbumin, alpha subunit and haptoglobin) serum concentrations of one or more were raised in 33% of patients with local disease and 81% of those with advanced breast cancer. However, marker concentrations were often only marginally raised, and are unlikely to provide sensitive guide to tumour burden. CEA, lactalbumin and alpha subunit were detectable in 68%, 43% and 40% respectively of extracts of primary breast cancers.


Serum IgG, IgM and slow alpha-globulin levels in carrageenan-treated rats.

Serum levels of IgM, IgG, slow alpha 1- and slow alpha 2-globulins were measured either by quantitative radial immunodiffusion (IgG) or immunoelectrophoresis (IgM and slow alpha-globulins) during the 3-week period after i.p. injection of 50 mg potassium carrageenan. There was a significant elevation in levels of IgM and slow alpha 1-globulin, maximal on Day 4 and returning to normal by Day 14. Slow alpha 2-globulin was detectable within 24 h, reached a peak at Day 2, and was no longer measurable in most rats by Day 14. Levels of IgG however, were unaffected by carrageenan injection.


Effects of bacterial endotoxin and corticosteroids on plasma concentrations of alpha 2 macroglobulin, haptoglobin and fibrinogen in rats.

Bacterial endotoxin injected into rats resulted in increased plasma concentration of alpha 2 macroglobulin, haptoglobin and fibrinogen. Cortisone acetate injected i.m. by itself was sufficient to increase the plasma concentration of haptoglobin by 54% and to a lesser extent the concentrations of the other two proteins. When cortisone acetate and/or cortisol succinate were injected simultaneously with varying doses of endotoxin, the effects of the corticosteroid differed for each plasma protein. Doubtless because of the effect of cortisone by itself the slope of the dose-response relationship for haptoglobin was greatly reduced. In contrast to this the slope for alpha 2 macroglobulin was reduced and that for fibrinogen was unaffected. These findings suggest that, if effects due to endogenous corticosteroids are to be avoided, increases in plasma fibrinogen will serve best as indicators of stimulation of the acute-phase response. Since, however, the relative increase of alpha 2 macroglobulin due to the lowest dose of endotoxin was much greater than that of fibrinogen, increases in concentration of the former protein represent the most sensitive indication of the acute-phase response. Consideration of the responses in individual rats has made possible division into those with more or less than average increases for all 3 plasma proteins and those showing irregular responses. Especially in the group which had received the lowest dose of endotoxin, a much larger number than would be expected on a random basis was found to respond regularly with either more or less than average increases for all 3 proteins.


Human blood and rabbit peritoneal leucocytes as sources of endogenous mediators.

"Buffy-coat" residues from human blood have been investigated as a source of leucocytic endogenous mediator (LEM). After separation from most of the erythrocytes and uptake of opsonized zymosan by the polymorphonuclear leucocytes (PML) and macrophages, the cells were incubated in Hanks' medium. To demonstrate the presence of LEM the supernatant thus produced was injected into rats and then after 24 h increases in the plasma concentrations of haptoglobin, alpha 2 macroglobulin and fibrinogen were estimated. For comparison rats were also injected with LEM prepared from rabbit peritoneal cells. Because these cells were obtained from rabbits which had been stimulated by i.p. injection of glycogen, addition of zymosan in vitro was not required. Despite the different species from which the leucocytes were obtained and the different method of preparation of LEM, similar increases in concentration of haptoglobin, fibrinogen and alpha 2 macroglobulin in the plasma of the recipient rats were obtained. Because injection of LEM resulted in very varied increases in concentration of each of the 3 plasma proteins, an attempt was made to ascertain whether positive responses for all 3 proteins occurred in the same rats or whether the rats responded randomly. The presence of endogenous pyrogen as a constituent of the LEM produced from both sources was confirmed using both rabbits and prewarmed mice (Bodel and Miller, 1976).


beta-Thromboglobulin, platelet production time and pletelet function in vascular disease.

Plasma beta-thromboglobulin (beta TG) levels were measured in 103 healthy controls and 112 patients suffering from either peripheral vascular disease (PVD), or cerebrovascular disease (CVD) or deep vein thrombosis (DVT). Plasma beta TG was significantly elevated in 46 PVD patients and 24 recent DVT patients compared to controls, but did not differ significantly in 18 chronic DVT and 24 old CVD patients. In addition, heparin neutralizing activity (HNA) and platelet aggregation induced by adenosine diphosphate, 1-epinephrine and thrombin were compared in 33 out of the 46 PVD patients to 33 controls. The mean HNA was significantly shorter in the PVD patients than in controls. The rate and extent of platelet aggregation were increased in PVD patients compared to controls, but the difference was not statistically significant. Platelet production time (PPT) was measured in 20 controls, 35 PVD patients, nine chronic DVT and 12 chronic CVD patients; significantly shorter PPT was only observed in 14 patients with advanced PVD compared to controls, suggesting increased platelet consumption in these patients. All four assays (plasma beta TG, HNA, platelet aggregation and PPT) were performed in 25 patients; no correlation between the four tests was found in these patients suggesting that the tests were measuring various aspects of platelet function. These results suggest that in vivo platelet consumption as well as platelet aggregation and 'release reaction' are presumably enhanced in PVD and recent DVT patients and that plasma beta TG and PPT assays may be better and more specific indicators of in vivo platelet activation than in vitro platelet aggregation test.


Studies on pregnancy-associated plasma protein A in the third trimester of pregnancy.

The plasma concentration of pregnancy-associated plasma protein A (PAPP-A) was measured in 34 women during the last 10 weeks of pregnancy. From 30 to 36 weeks the concentration of this protein increased steadily. Thereafter the concentration of PAPP-A rose more steeply, the highest amounts being found in early labour. The concentration of PAPP-A in peripheral  venous blood and in the uterine vein was much the same. It was less in the retroplacental blood and a great deal less in the peritoneal fluid. The day-to-day variation was small; the coefficient of variation at 38 weeks was only 7.3 per cent. After delivery, the concentration of PAPP-A fell more slowly than other placental proteins and steroids, the average half-life being 51 hours. Although there is no doubt that PAPP-A is a product of the syncytiotrophoblast, our findings suggest that it is not simply secreted by the chorionic villi directly into the intervillous space but makes its way into the maternal circulation by a more circuitous route.


The relation between the concentration of placental specific proteins in retroplacental and peripheral blood.

Concentrations of four placental proteins: human placental lactogen (hPL), placental protein 5 (PP5), pregnancy specific beta 1 glycoprotein (SP1) and human chorionic gonadotrophin (hCG), and a normal serum component, alpha 2 macroglobulin, were measured in the peripheral circulation and in blood obtained from the retroplacental space in 20 women at term delivery. Levels of hPL and PP5 were higher in the retroplacental blood than in the peripheral circulation in all patients. By contrast, levels of SP1 and hCG were consistently lower in retroplacental blood than in the peripheral circulation. Similarly, levels of alpha 2 macroglobulin were lower in the retroplacental blood. It is suggested that this 'reverse' gradient is a technical arterfact. These findings are discussed in relation to synthesis of placental proteins in a site distal to the retroplacental space, and the introduction of a technical artefact in the collection of samples.


Effect of defaunation on the metabolism of rumen micro-organisms.

1. Rumen contents of a fasted fistulated wether, obtained in a faunated, defaunated and refaunated period were incubated in vitro with a mixture of cellobiose and maltose, in the presence of ammonium bicarbonate and 32PO43-. Total synthesis of microbial N (Nt) was calculated from 32P incorporation and N:P determined in microbial matter. The N:P value was not affected by defaunation. Net synthesis of microbial N (Nn) was calculated from ammonia-N incorporation. An estimate of degradation of microbial N was calculated as Nt-Nn. Energetic efficiency of synthesis was calculated from the volatile fatty acids produced during incubation, as g N incorporated per kg organic matter fermented (g N/kg OMf). 2. Defaunation decreased the proportions of acetate, butyrate and methane and increased those of propionate in fermentation end-products. Fermentation rate when expressed per mg microbial N was not affected by defaunation. 3. Expressed per unit volume of rumen contents, Nn was increased by defaunation whereas Nt remained unchanged. Thus, a decrease in degradation can be calculated. Energetic efficiences of total and net synthesis were increased from 35 and 13 to 47 and 30 g N/kg OMf respectively. 4. Specific rates of both total and net synthesis of microbial N were significantly increased by defaunation whereas the specific rate of degradation was not affected.


Studies on the activation of canine trypsinogens in vitro.

The activation of canine anionic and cationic trypsinogen by enterokinase, trypsin, thrombin, plasmin and extracts from canine granulocytes were studied in vitro. Enterokinase activates both trypsinogens about 1000 times faster than trypsin. The enterokinase-catalyzed activation is not inhibited by the main serum protease inhibitors, alpha-macroglobulin and alpha 1-antitrypsin. alpha-Macroglobulin cannot inhibit the activation of the trypsinogens by trypsin but this reaction is completely inhibited by alpha 1-antitrypsin. The results are discussed in relation to the pathogenesis of acute pancreatitis.


Rapid removal to the liver of intravenously injected lipoprotein lipase.

Lipoprotein lipase was purified from bovine milk and labeled with 125I. After intravenous injection to rats the labeled lipase rapidly disappeared from the blood. The initial half-life was about 1 min and more than 70% of the radioactivity was found in the liver at 10 min. 30 min after the injection about 10% of the injected radioactivity was present in acid-soluble form in blood, indicating that the enzyme had been rapidly degraded. Injection of asialofetuin, ribonuclease B or mannan in amounts known to block the hepatic receptors for glycoproteins with exposed galactose, N-acetylglucosamine or mannose residues did not retard the removal of the lipoprotein lipase. Thus, some other, as yet undefined, receptor is implicated. Lipoprotein lipase is known to bind to heparin and some related polysacchrides. Heparin injected before the enzyme delayed its removal and heparin injected after the enzyme caused an immediate increase in blood radioactivity, signifying return from tissues to blood of labeled enzyme. Lipoprotein lipase is present at the endothelium in several extrahepatic tissues and is rapidly turned over. Its presence in blood in appreciable amounts would cause a derangement of lipid transport. The efficient hepatic removal of the enzyme may thus serve an important physiological purpose in keeping the blood levels of this enzyme low.


Blood transfusion and renal transplantation.

Blood transfusions given to potential renal transplant recipients are beneficial to subsequent graft survival. This study has shown an association graft survival. This study has shown an association between the inhibitory activity of patients' plasma on their own non-specific lymphocyte responses to antigen and the administration of blood transfusions. Most of the inhibitory activity was related to alpha-2 macroglobulin. Alpha-2 macroglobulin was able to prolong the survival of rat cardiac allografts. Immunoregulatory activity related to the alpha globulins of plasma has been described in a wide variety of conditions (i.e. cancer, pregnancy, diabetes) and this study has shown similar activity following blood transfusions.


An electron microscopic study of neurones in the lateral geniculate nucleus of the monkey labelled by retrograde transport of horseradish peroxidase.

In a previous report horseradish peroxidase (HRP) reaction product has been described as accumulating in a restricted part of neurones in the lateral geniculate nucleus following injections of HRP into the visual cortex of the primate. With the aid of long series of ultrathin sections from the same material labelled neurones have been partially reconstructed in the present study, and it has been confirmed that HRP reaction product aggregates as a group of electron-dense lysosome-like bodies which form a cap to one side of the nucleus.


Reduction by reserpine of the accumulation of retrogradely transported [125I]nerve growth factor in sympathetic neurons.

Experiments were carried out to determine if stimuli which augment preganglionic nerve activity to sympathetic neurons, and thereby cause trans-synaptic induction, increase the retrograde transport of nerve growth factor (NGF). It was found that nerve activity had no effect on retrograde transport of [125I]NGF. It was found, however, that reserpine decreased retrograde transport of [125I]NGF and this inhibition was characterized. Reserpine decreased the maximal accumulation of intravenously administered [125I]NGF in superior cervical ganglia (SCG) by about 60%. It also caused a distinct shift in the time course of accumulation so that maximal accumulation was seen 12 h after [125I]NGF injection rather than at 9 h as in control animals. Reserpine had no effect on retrograde transport in sensory neurons. Dose--response curves showed that maximal inhibition occurred with doses of reserpine of 2.5 mg/kg i.p. and that reserpine was not able to completely block transport at any dose. The maximal inhibition of retrograde transport was achieved within 30 min of reserpine administration and inhibitory activity was unchanged for 36 h. The ability of sympathetic neurons to transport [125I]NGF subsequently recovered and was normal 96 h after reserpine administration. The inhibitory effect of reserpine appears to be due to an action at or very near to the nerve terminal since it was effective at reducing NGF transport at very low doses (0.33 microgram) when co-administered directly into the eye with [125I]NGF. An action of reserpine at the nerve terminal was further suggested by the inability of reserpine to affect transport if the drug was given 4 h after [125I]NGF administration. Based upon these data, it is suggested that there may be two pools of retrogradely transported NGF and that only more rapidly turning over pool is reserpine-sensitive. This pool may represent the retrogradely moving synaptic vesicles or some derivative of the vesicles.


Extra-axonal diffusion in the rabbit optic system: a caution in axonal transport studies.

The hazards of using optic nerve (as opposed to optic tract and more distal components of the optic system) to study axonal transport were highlighted by observing the fate of [14C]serine and [3H]glycerol injected into the rabbit eye. Despite prior blockage of axonal transport with colchicine, appreciable radioactivity rapidly appeared in the optic nerve adjacent to the injected eye. Radioactivity decreased exponentially along the entire optic chiasm. Counts were distributed among the lipid, protein, and acid-soluble fractions. Separation of optic nerve lipids revealed appreciable labeling of most lipid classes including those characteristic of myelin; a markedly different labeling pattern was observed for axonally transported lipids. The data are consistent with a mechanism involving extra-axonal diffusion of precursor into the surrounding glia followed by incorporation into lipids and proteins of those cells and ultimately myelin. The phenomenon is discussed in relation to possible errors that were made in interpreting earlier experiments.


[Contribution of new methods of quantitative cytology and immunology to the understanding of lymphoproliferative syndromes].

Seventy one patients with a lymphoid hemopathy, three with agammaglobulinemia and six normal controls were investigated with regard to their blood lymphocyte membrane-associated light chains. Detection and quantitation of antigenic determinants were performed by means of peroxidase-labeled antibodies. Compared to normal controls, values found in chronic lymphocytic leukemia (CLL) were very low (tenfold decrease). The number of antigenic determinants on lymphoid cells from patients with blast crises supervening in CLL, prolymphocytic leukemia, Waldenstrom's macroglobulinemia and Burkitt cell acute leukemia were significantly higher than those seen in patients with CLL. The data obtained in this investigation through quantitative immunocytology constitutes a new parameter for the classification of lymphoid hemopathies and for an approach to their pathogenesis, in particular if the quantity of membrane immunoglobulin correlates with the stage of cell maturation.


["Non-specific cross reacting antigen (NCA)", a tracer of human granulocytes and monocytes].

The non specific cross reacting antigen, or NCA, is a normal tissue antigen that cross reacts with CEA. It bears a specific antigenic determinant that is absent from CEA. Immunocytological studies first pointed out that NCA, but not CEA, is present in alveolar macrophages and polymorphonuclears. Further work demonstrated that NCA is a marker of granulocytic series: it appears at the stage of promyelocyte and likely is linked to the azurophilic granules. The same antigen is also present in peripheral blood monocytes, it is not detectable in them, but after adherence to glass. The possible role of NCA in these lytic enzyme rich cells is discussed.


Topical application of epinephrine during bronchoscopy in barbiturate-halothane-anaesthesia and its influence on cardiac action.

Since the introduction of halothane as an anaesthetic there have been several reports about cardiac arrhythmias during application of epinephrine in combination with halothane. This complication was also reported when epinephrine was used as local haemostyptic. We describe the effect of topical epinephrine upon cardiac action during bronchoscopy in barbiturate-halothane-anaesthesia. 21 patients with an average age of 66.4 years underwent bronchoscopy mainly because of suspect bronchial carcinoma, they were monitored by lead I ECG and blood gas analysis was carried out repeatedly. Immediately following biopsies, sponses with epinephrine solution 1:1000 were applied to the bronchial mucosa - the average epinephrine dose of these sponges was 0.12 mgs as calculated before by weighting. During this procedure we could not observe significant cardiac arrhythmias following epinephrine application. We conclude this to be the result of limited dosage and maintenance of adequate ventilation and oxygenation. Thus, epinephrine can be used as a local haemostyptic during bronchoscopy in halothane anaesthesia without significant risk when the above described precautions are observed.


Place of immunological examinations in extrinsic asthma.

The provocation test is - in spite of its drawbacks, especially the risk for the patient - the most reliable way to identify causative agents in extrinsic asthma. The immunological genesis of the obstruction provoked must be proved with adequate methods however. RAST (Radio-allergo-sorbent-test) and histamine liberation are valuable in vitro techniques for the identification of atopic sensitizations. Their disadvantages - the expensive equipment and the time-consuming procedure respectively - limit their widespread use. A carefully performed and evaluated prick test possesses a comparable specificity with acceptable risk. Less satisfying is the situation in nonimmediate asthma. Precipitations tests are at present most valuable. Main indications for immunological methods in extrinsic asthma are: - the identification of causative allergens and the kind of underlying immune reactions; - the control and - in the future - the individualization of hyposensitization procedure. The future use of allergen extracts containing a minimum of non-specific irritants is extremely important.


Ductular carcinoma of the breast: serum antibodies to tumor-associated antigens.

Serum antibodies to tumor-associated antigens of breast carcinoma have been studied by indirect immunofluorescence in 109 patients with breast carcinoma and 125 controls, including age/sex matched normal individuals, patients with nonmalignant disease, and patients with malignant disease other than breast cancer. We report here that sera of a large proportion of patients with ductular carcinoma of the breast have antibodies to cell surface and/or intracellular antigens of autologous tumor cells and include evidence that the antigens are absent from a considerable range of normal and other types of malignant tissues. In addition to testing of control sera, specificity of the reacting antibodies was investigated further by testing of sera with normal breast tissue and the absorption of sera from breast cancer patients with various normal tissues and cancer cells. The significance of the findings in breast cancer is discussed.


Purification and biochemical characterization of a virus-specific reverse transcriptase from human osteosarcoma tissue.

A RNA-dependent DNA polymerase (RTase) was purified from human osteosarcoma tissue by successive column chromatography of the microsomal fraction on DEAE-cellulose (DE-23 and DE-52) and phosphocellulose. The purified enzyme has a molecular weight of about 68,000, a pH optimum of 8.1, a Mg2+ optimum of 0.8 mM, Mn2+ optimum of 1.0 mM and a KCl optimum of 60 mM. The enzyme transcribes (rA)n . (dT)12, (rC)n . (dG)12-18 and (2-O-methyl C)n . (dG)18, but is unable to transcribe (dA)n . (dT)10. The enzyme has no catalytic activity in the presence of oligodeoxynucleotide initiators alone, indicating the absence of terminal deoxynucleotidyl transferase. The purified enzyme is able to transcribe the heteropolymeric regions of a 70S RNA from R(Mu)LV. The presented data support the presence of a RNA-dependent DNA polymerase in human osteosarcoma tissue with biochemical properties, resembling those of C-type RNA tumor viruses.


Serological characterization of a purified reverse transcriptase from osteosarcoma of a child.

Serological analysis of the reverse transcriptase (RTase), purified from human osteosarcoma tissue, has shown that it is antigenically related to DNA polymerases from BEV and from RD-114. No cross-reactivity of the osteosarcoma RTase was observed with RTases purified from AMV, RLV, SiSV, GaLV and from human spleen of a patient with myelofibrosis.


Suramin: a potent inhibitor of the reverse transcriptase of RNA tumor viruses.

Suramin--a well-known antitrypanosomal agent--was found to exert a strong inhibitory effect on the RNA-directed DNA polymerase (reverse transcriptase) activity of several oncornaviruses such as Moloney murine leukemia virus, murine Rauscher leukemia viruses, Moloney murine sarcoma virus and avian myeloblastosis virus. Inhibition of enzyme activity was obtained with both endogenous viral RNA and (A)n . oligo(dT) as the template-primer. Suramin effected a 50% inhibition of the reverse transcriptase activity of oncornaviruses at a concentration range of 0.1--1 microgram/ml. In this aspect it compared favorably to ethidium bromide, another trypanocide drug which is considered as one of the most powerful inhibitors of oncornaviral DNA polymerases. The inhibition of reverse transcriptase activity by suramin was competitive with the template-primer, (A)n . oligo(dT), suggesting that the drug may interact with the template-primer binding site of the enzyme.


A Markham rotation enhancement of two polygon types from isolated negatively stained human placental coated vesicles.

A simple image enhancement method has been used to reinforce the rotational symmetries of two types of polygon which form the facets of the clathrin lattice of human placental coated vesicles. Hexagons and pentagons were found but no evidence of other polygons (up to enneagon) was obtained.


Vitamin D binding globulin phenotypes in liver disease.

The binding properties towards vitamin D metabolites of plasma from individuals with the three common Gc-globulin phenotypes, Gc-1, Gc-2 and Gc-2-1, have been found to be identical. In patients with liver disease there is a good correlation between the levels of Gc-globulin andalbumin in plasma. In addition the Gc-globulin levels correlate well with the ability of plasma to bind 25-hydroxycholecalciferol. Patients with the Gc-2-1 phenotype showed a significantly smaller depression in plasma Gc-globulin than those with the Gc-2 and Gc-1 phenotypes. The relation of these findings to the pathogenesis of disorders of calcium metabolism in liver disease is discussed.


A nephelometric study of the reaction of monoclonal rheumatoid factor with heat aggregated gamma globulin and sera from patients with immune complex diseases.

Monoclonal rheumatoid factors (MCRF) have previously been used in a variety of assays for the detection of IgG-containing circulating immune complexes. We have isolated a MCRF from a patient with a lymphoproliferative disorder and have used a nephelometric technique to characterize its reaction with heat-aggreagated gammaglobulin (HAGG) used as a source of artificial immune complexes. The method is simple, economical and rapid and will detect as little as 6 microgram/ml of HAGG over a wide range of physicochemical conditions. A clinical study demonstrated that the sera from thirty-five out of fifty-eight patients (59%) with rheumatoid arthritis and twenty-one out of seventy-four patients (28%) with systemic lupus erythematosus (SLE) gave increased precipitation with MCRF compared with 232 blood donors. However, in marked contrast to previous studies, sucrose gradient ultracentrifugal analysis of nine strongly precipitating sera revealed that in eight the MCRF precipitated with material sedimenting in the monomeric IgG position. In only one specimen did the MCRF react with material sedimenting in heavier regions. It is suggested that different MCRFs vary in the specificity for binding IgG complexes and these reagents should be carefully characterized before becoming established in nephelometric assays for circulating immune complexes.


Demonstration of two disease specific antigens in circulating immune complexes.

A method is described to assess antigenic cross-reactivity between soluble immune complexes precipitated from sera with polyethylene glycol. The precipitated complex from one serum was dissociated in acid and used to coat a plastic cup. Radioiodinated complexes from another serum were dissociated in the cup, neutralized and allowed to reassociate overnight. The binding of the labelled complex was used to measure the cross-reactivity between the complexes. Using this technique, complexes from a group of patients with haematuria and hypertension have been found to share an antigen, and a different antigen was found in patients with bullous pemphigoid. The participation of rheumatoid factors in the cross-reactions is unlikely, and no cross-reactivity of either group was found with sera from patients with rheumatoid arthritis.


Comparative studies on the expression of Fc-receptors, Ia antigens, and beta 2-microglobulin following in vitro herpes simplex virus infection of human lymphoid cells.

Characteristics of the specificity and affinity of surface Fc-receptors induced on Raji (a Burkitt's lymphoma derived B cell line) and Molt-4 (a T cell line derived from a leukaemic patient) after herpes simplex virus (HSV) infection were studied, together with the expression of other surface markers. Inhibition experiments of EA-rosetting suggested that Raji cells infected by HSV had a higher affinity for soluble aggregated IgG than for IgG serologically adsorbed onto erythrocytes; an opposite pattern was noted with Molt-4, a non-Burkitt's lymphoma cell line. Using radiometric and microspectrofluorometric techniques, the expression of Fc-receptors, as detected by their capacity to fix aggregated IgG, paralleled the expression of serologically detectable Fc-receptors and Ia determinants. In contrast to beta 2-microglobulin, both Fc-receptor and Ia antigen expression increased following Hsv infection: this suggests a specific induction of these markers on human lymphoid cells following HSV infection. To our knowledge, this is the first report showing a specific induction of both Fc-receptors and Ia antigens by HSV following in vitro infection of human lymphoid cells.


Concealed extrasystoles due to Wenckebach conduction delay within the reentry loop.

Long electrocardiographic strips were analyzed from an aged patient whose heart rhythm had periods of unifocal ventricular extrasystoles with fixed coupling intervals. Periods of gradual prolongation of the coupling interval finally led to omission of a ventricular premature beat. This sequence was repetitive and is considered to be the results of reentrant extrasystoles with a 3:2 Wenckebach type of conduction delay within the reentry loop. The mechanism of concealed conduction due to overlong propagation within the reentry loop is discussed.


Skeletal changes in Refsum's disease.

Refsum's disease is a rare inherited disease of lipid metabolism. The cardinal diagnostic features include polyneuritis, cerebellar ataxia, an atypical pigmentosa and a high CSF protein. The disorders is accompanied by the accumulation in the tissues, especially the liver and kidneys, of the lipid 'phytanic acid'. The disease is due to the absence of the enzyme phytanic acid alpha-hydroxylase which catalyses the conversion of phytanic acid to alpha-hydroxy phytanic acid the initial step in its further metabolism. In his original monograph Refsum (1945) documented a number of skeletal abnormalities and the full spectrum of changes that occur has become clear though the accumulating subsequent reports. This paper documents the osseous changes in three members with the disease in a single family. These include epiphyseal dysplasia, especially pronounced in the knees, and shortening and deformity of many of the tubular bones in the hands and feet.


Virus-like particles and macromolecules in human milk and breast tumors.

Relevant data pertaining to present evidence for virus-like particles and virus-related macromolecules in human milk and breast tumors are presented. A critical review and discussion of reported observations concerning virus-related macromolecules will include RNA-directed DNA polymerase, viral antigens, and RNA related to murine mammary tumor virus and/or Mason-Pfizer monkey virus. From the standpoint of clinical applications, the finding of viral-related antigens in human breast tumors and evidence for specific host immune responses to one or more of these antigens may be especially pertinent. The latter data, therefore, will be discussed in depth as to possible employment of these parameters in diagnosis, prognosis and possible management of the human disease.


Cutaneous manifestations of acute carbon monoxide poisoning.

A patient with acute carbon monoxide poisoning due to leakage of gas from the exhaust system in his automobile noted edema and blister formation over large areas of the skin of one half of his body. Rhabdomyolysis, acute but transient renal insufficiency, and hemolytic anemia developed subsequently.


Procainamide-induced sinus node dysfunction in patients with chronic renal failure.

Two patients with chronic renal failure developed transient sinus node dysfunction requiring insertion of a temporary pacemaker while receiving procainamide to control ventricular arrhythmias. Blood levels of procainamide were found to be elevated, although at these levels, sinus node dysfunction has not previously been reported. Following discontinuance of procainamide, sinus rhythm returned. A combination of factors, including elevated levels of N-acetyl procainamide, the metabolite of procainamide with anti-arrhythmic properties, are suggested as possible contributory causes for the ECG findings. Thus, procainamide may produce electrophysiologic features of "sick sinus syndrome" in patients with chronic renal failure even when blood levels of this substance are being monitored.


[Circular transhepatic drainage as a palliative surgical measure in central bile duct obstruction].

A transhepatic drainage tube was used in 27 patients suffering from echinococcus alveolaris of the liver of malignant process of the portal fissure from 1967 to 1978. If an hepatocholangioenterostomy is not possible, another surgical palliative procedure is necessary for drainage of the bile. The transhepatic drainage tube is a simple palliative method. The technique of this procedure, the indications, the possible complications, and the advantages are reported.


Antiviral agents: action and clinical use.

The development of antiviral agents has been hindered by a variety of problems. There are fundamental biological differences between viruses and other infectious agents. Viruses are strictly dependent on cellular metabolic processes and possess very limited intrinsic enzyme systems and building blocks which may serve as targets for drugs. Antiviral drugs must also possess the ability to enter the host cell. Viral replication consists of a series of events, each of which can be interfered with, leading to interruption of the viral replication cycle. Currently, the major antiviral agents in therapeutic use are amantadine, idoxuridine and vidarabine. Methisazone and isoprinosine are also used in some areas. Immunoglobulins have some antiviral activity. Immune serum globulin and high titred hepatitis B immune globulin have both been used in prophylaxis of viral hepatitis. However, studies in this area have not been well controlled and results in some areas are conflicting. Interferon appears to be the most exciting antiviral agent yet discovered. However, its potential is limited by its availability, which remains dependent on biological method. Significant progress has been made recently, though, which may lead to the chemical synthesis of interferon and thus to an antiviral agent active against many viruses.


The value of plasma TSH estimation and TRH stimulation test in the course of treatment of differentiated thyroid cancers.

Plasma TSH levels were measured on 114 occasions in 96 patients treated for differentiated thyroid cancer. Prior to thyroid surgery, plasma TSH levels were within the range of normal. Plasma TSH levels increased slightly following partial thyroid resection and definitely after total thyroid ablation. In patients where the removal of normal thyroid induced hormonogenesis in thyroid tumours, plasma TSH levels were dependent on the hormonal secretion of the tumour as shown by inverse relationship between TSH and both PBI and 131I uptake. The increase of radioiodide uptake following stimulation by exogenous bovine TSH was inversely related to the plasma thyrotropin levels. The suppressibility of enhanced thyrotropin levels was complete with individually adjusted doses of synthetic thyroid hormones. With the exception of patients on suppressive treatment, TRH administration induced increase in plasma TSH levels. The findings are discussed with regard to the role played by TSH in the induction of hormonogenesis in thyroid tumours. The practical values of TSH estimation and TRH stimulation seem to be low; the measurements of thyrotropin levels may be important for the estimation of the suppressive effect in the course of and following withdrawal of treatment with thyroid hormones.


Effect of the pesticides phosalone and lindane on the activity of some dipeptidases and disaccharidases in rat intestinal mucosa.

The intestinal dipeptidase and disaccharidase activities were investigated in 120 male albino rats of the Wistar strain after administration of 21 mumol.kg-1 body weight phosalone, 14.8 mumol.kg-1 body weight lindane and 10.5 mumol.kg-1 body weight phosalone combined with 7.4 mumol.kg-1 body weight lindane. The dipeptidase activity under the effect of these comparatively low doses of pesticides reveals slightly to moderate changes. The activity of intestinal disaccharidases after a 90-day phosalone and lindane treatment is markedly decreased, particularly that of sucrase. The mechanism of these changes remains unknown.


A reappraisal of palliative surgery for total anomalous pulmonary venous connection: description of a new operative technique.

The need for effective palliation of critically ill infants born with total anomalous pulmonary venous connection is reappraised. Careful study of autopsy specimens in patients with this cardiac malformation (patient ages ranging from newborn to 5 1/2 mth) revealed 4 favorable anatomic dispositions in 16 selected hearts: (1) a horizontal vein or a lobar vein larger than 3-mm size was present in every case; (2) these veins were of adequate length for a shunt anastomosis; (3) the heart could be rotated for easy access to the left atrium; and (4) one of the two separate connecting veins in the mixed type could be utilized. Based on morphologic observations and trial procedures on the autopsy specimens, a new palliative operation is proposed which can be performed under normothermia and without cardiopulmonary bypass.


Helper activity of T cells stimulated in long-term culture.

Thymus-derived (T) cells obtained from three categories of in vitro antigen-induced proliferative responses were assayed as sources of helper T cells. These categories are exemplified by (a) direct stimulation of keyhole limpet hemocyanin (KLH)-primed cell with KLH, which results in high indexes of proliferation; (b) direct stimulation of apoferritin-primed cells with apoferritin, which does not result in indexes of proliferation above background levels; (c) trans-stimulation of unprimed cells with X-irradiated KLH-primed cells which results in indexes of proliferation comparable to category (a). Our results indicate that levels of [3H]thymidine incorporation by proliferating populations are not an accurate reflection of helper T cell generation. Directly stimulated KLH and apoferritin-primed cells give rise to highly enriched populations of antigen-specific helper T cells which support both IgM and IgG antibody responses in vitro. Moreover, these specific helper T cells are functionally restricted by products encoded by the I region of the major histocompatibility complex (MHC). Helper T cells generated in KLH-trans-stimulated cultures are not KLH-specific in that comparable levels of helper activity are expressed using either KLH or apoferritin as carriers. These non-KLH-specific helper T cells only support the production of IgM antibody in vitro and they are not functionally restricted by MHC products.


Amino acid sequence analysis of group e allotype-related peptides derived from lagomorph IgG.

A tryptic peptide (T 19) was isolated from a CNBr fragment of IgG from several representatives of present-day lagomorphs including domestic rabbit, hare, cottontail rabbit and pika. The amino acid sequence of this peptide revealed differences which can be correlated with serological polymorphism associated with group e IgG allotypes. The results confirm previous studies establishing the importance of Ala309 for the expression of e 15, particularly the i subspecificity. Arg315 correlates with the expression of the j subspecificity.


Generation of virus-specific cytotoxic T cells in vitro. I. Induction conditions of primary and secondary Sendai virus-specific cytotoxic T cells.

H-2-restricted cytotoxic T cells specific for Sendai virus were generated in vitro in a primary response from normal mouse lymphocytes cultured in the presence of infective as well as inactivated Sendai virus. Antigen-presenting cells of different origin, including T cells, were found to be effective stimulators. Antibodies to Sendai virus were shown to inhibit the activation of specific precursor killer cells when added to cultures before, but not after, the addition of viral antigen. Data obtained by Lyt phenotyping, revealed that precursor killer cells specific for Sendai virus reside in the Lyt-2,3+ T cell population and that Lyt-1,2,3+ T cells are not required for the generation of cytotoxic lymphocytes. Different activation kinetics were demonstrated for primary and secondary antiviral cytotoxic responses, and the analysis of the proliferation and stimulation requirements suggests qualitative differences.


N-acetylprocainamide and ischemia-induced ventricular fibrillation in the dog.

Open-chest dogs anesthetized with pentobarbital were treated with saline or N-acetylprocainamide (20 mg/kg, i.v.) 10 min prior to simultaneous ligation of the left anterior descending and septal coronary arteries. Ventricular fibrillation occurred in 20 of 26 control dogs but in only 6 of 15 dogs treated with N-acetylprocainamide (P less than 0.05). Since N-acetylprocainamide significantly reduced spontaneous heart rate this may have contributed to its antifibrillatory effect.


Comparative evaluation of 7 helminth antigens in the enzyme-linked immunosorbent assay (E.L.I.S.A.).

112 sera from Europeans with parasitologically proven helminthiasis were tested in the enzyme-linked immunosorbent assay (E.L.I.S.A.) against 6 crude extracts of various helminths (2 of adult worms: Dipetalonema viteae, Fasciola hepatica; 3 of eggs: Ascaris suum, Toxocara canis, Schistosoma mansoni; and of Echinococcus granulosus scolices) and against bovine hydatid fluid. Each serum was tested simultaneously at a fixed dilution of 1:160 against all antigens. Extensive cross-reactions were observed, leading to the conclusion that non-purified helminth antigens, even in combination, are of limited value for reliable serodiagnosis in E.L.I.S.A.


Pepstatin, an inhibitor of acid kininogenases and ascites retardant in neoplastic disease.

Evidence is reviewed that pepstatin, an inhibitor of acid kininogenases such as cathepsin D, may be an effective therapeutic agent in retarding ascites accumulation in certain cancers. The evidence for this conclusion is based on the actions of pepstatin in retarding ascites in six different tumor strains inoculated into various species of mice, as well as the demonstration that cathepsin D activity is reduced in vivo in several organs following pepstatin administration. The latter is significant since we have postulated that ascites formation, in good part, is due to leukokinin formation, which is catalyzed by cellular-released cathepsin D.


Neurobehavioral performance of low-birthweight infants at 40 weeks conceptional age: comparison with normal fullterm infants.

This study compares the neurobehavioral status of 118 low-birthweight infants tested at 40 weeks conceptional age with that of 76 normal fullterm infants. A neonatal neurobehavioral examination comprising 21 test and four summary items was used. The most striking differences between the groups were found in visual and auditory orienting, with approximately two-thirds of the low-birthweight infants falling below the range of performance of the fullterm group. Items testing motor performance showed a lower incidence of deviant performance among the low-birthweight infants. Of the 21 test items, 19 could be assigned cut-off scores, below which infants can be considered deviant on the specific items.


Progressive rubella panencephalitis: clinical course and response to 'isoprinosine'.

This report describes the clinical course of a 16-year-old Canadian-born Chinese boy who has progressive rubella panencephalitis. The progression, the lack of response to 'Isoprenosine', and the additional finding of myopathy are discussed. The clinical and pathological features of this rare, progressive neurological disorder are also summarized. In view of the last major rubella pandemic occuring in the mid-60s, it is likely that during the next few years physicians will diagnose an increased number of patients with this distinct neurological entity.


Combination chemotherapy with a new anthracycline glycoside, aclacinomycin-A, and active drugs for malignant lymphomas in P388 mouse leukemia system.

Aclacinomycin-A is a new anthracycline glycoside and has less cardiotoxicity than adriamycin. In an attempt to provide an experimental model of a phase III study of aclacinomycin-A, particularly for the treatment of malignant lymphomas, various therapeutic designs of combinations of this drug with other conventional agents were investigated using a P388 mouse leukemia system. Aclacinomycin-A showed no treatment schedule dependency in this tumor system and the optimal dosage of this drug was twice higher than that of adriamycin on each treatment schedule; i.e., single treatment on day 1, three treatments on days 1, 5, and 9, or 10 treatments on every other day from days 1 to 19 after an inoculation of 10(6) leukemic cells on day 0. This antibiotic was ineffective against an adriamycin-resistant subline of P388 leukemia. Among combinations of aclacinomycin-A with cyclophosphamide, vincristine, procarbazine, or bleomycin, the combinations of aclacinomycin-A with cyclophosphamide or vincristine showed a therapeutic synergism in P388 leukemia system.


Specificity of micro-leucocyte adherence inhibition test in patients with gastric cancer.

Specificity of micro-leucocyte adherence inhibition (LAI) test was studied in patients with gastric cancer. Because of the presence of nonspecific reactions, specificity appeared obscure unless interaction analysis was applied to the results of LAI tests. Application of interaction analysis revealed that only leucocytes from patients with gastric cancer showed specificity to gastric cancer antigen. Further technical improvement is required to eliminate nonspecific reactions which seem to be due to either leucocytes or tumor antigens.


[Simplification of the Papanicolaou stain which is easily reproducable (author's transl)].

A simple and well reproducable modification of the Papanicolaou stain is reported. The most important modification is the replacement of the natural stain hematoxyline by the qualitively excellent synthetic stain thionin which is immediately added to the alcoholic fixation solution for a fixation stain. The high labor intense alternation between alcohol water and alcohol is thus eliminated. The quality of the nuclear staining is very good. The fixation staining with thionin without counter-staining of the cytoplasma may be used for a rapid thionin stain which gives sufficient cytoplasmatic staining by the thionin for a rapid cytological smear for cancer. The counter-stain of the cytoplasma was also simplified. The over-all staining of this simplification of the pap smear is comparable to that of the original papanicolaou stain. This modification of the papanicolaou stain fulfills requirements which a stain in competition with the original papanicolaou stain should have. 1. The modification is qualitively equal to the original method, 2. the staining method is simpler, 3. the staining method is well reproducable, 4. the staining method is more economical.


[The early diagnosis of teratogenic malformations (author's transl)].

A case of anencephaly is reported which was diagnosed at 32 weeks gestation by ultra-sound despite a family history: AFP, HPL and estriol were normal and the case was referred late. It is suggested that all pregnancies require an ultra-sound scan between 16 and 20 weeks to rule out major fetal anomalies.


[Genetic conditions for the induction and realization of the action of specific T-suppressors immune to H-2 antigens].

Spleen T-cells of mice immunized intravenously with gamma-irradiated allogenic lymphoid cells at a high dose are shown to inhibit the activation of DNA synthesis in MLC. The difference in the whole H-2 complex or in either of its K, I, D regions or I-subregions is required and sufficient for induction of specific T-suppressors. The magnitude of the suppression is a sum of suppressive reactions induced separately by each of the immunizing complex components, the products of one of the H-2 regions (subregions) as shown by changing the source of stimulator cells in MLC. These components have a similar impact in the suppressor activity. Unlike the other T-cell subclasses, T-suppressors react only to serologically defined mutant antigens of the Dd allele, but do not react to the mutants of the Kb allele and do not discriminate Kb and Kba antigens. Specific suppressor T-cells recognize the I-subregion products without their linkage to the K/D region products. For elicitation of the suppression two conditions are required and sufficient: a) direct contact of immune suppressors with the corresponding antigenic determinant; b) the subregion IC identity between the suppressors and the responder cells. Intensity of suppression is reduced if F1 hybrid responder cells interact with parent suppressor cells, but is not reduced in the inverse arrangement of the experiment. Specific suppressor T-cells are presumed to differ from other subclasses of T-cells and to be similar to B-cells relatively to the nature and multiplicity of recognizable individual products of the H-complex and to narrow specialization of reactive clones.


Community residential treatment for skid row alcoholics.

A significant factor affecting the chance for recovery is the ability of the skid row alcoholic to form and sustain a relationship with someone or something. The author describes an open-ended, community residential program that encourages such relationships and which is proving to be an effective treatment model.


[The coagulation and fibrinolysis activities of the blood during various phases of labor].

In a group of 20 women 22 parameters of the blood coagulation and fibrinolysis systems were separately determined during spontaneous delivery. An increased activity of the coagulation factors was observed except of the factors V, IX and XIII. Factor VIII activity in plasma increased slightly during stage II of labour, probably related to stress. Fibrinogen as well as the factors II, VII and X exhibited congruently slight post partum decrease, which can be interpreted as the result of an increased turnover of these factors. Plasminogen levels as well as antiplasmin activity increased; the total fibrinolysis activity was not significantly changed. The increase of fibrin-fibrinogen degradation products, however, indicated the occurrence of discrete fibrinolytic or fibrinogenolytic reactions. The thrombelastograms demonstrated a shift of the balance of pro- and anticoagulatory factors in favour of an increased blood coagulation activity during labour.


[Gamma globulin in secondary antibody deficiency syndrome].

12 patients with secondary antibody deficiency syndrome received a new gammaglobulin compound intravenously (lyoph. SRK Sandoz). The drug was given in a 3% NaCl-solution once to four times. At the end of the infusion of 6 g gammaglobulin and after one week, serum levels were significantly higher. Gammaglobulin intravenously was well tolerated in all cases. Severe infections during the period of treatment were not observed.


A simple, versatile, sensitive and volume-independent method for quantitative protein determination which is independent of other external influences.

A method is described which in principle is a quantitative "spot analysis". 0.5-5 microliter of a protein solution in the concentration range between 0.01-10 mg/ml is used for one determination. The sample is taken up by capillary attraction in a 0.5-, 1-, 2- or 5-microliters capillary and transferred to a cellulose acetate strip. The protein is fixed and stained simultaneously by dipping the cellulose acetate strip into a solution of Amido Black or a benzoxanthene derivative (Hoechst 2495) dissolved in in methanol/acetic acid. After elution of the excess of dye (3 x 5 min) the quantitative evaluation can be performed in different ways: 1) The sample is fixed and made transpartent by incubation in dioxane/1-butanol and evaluated densitometrically (Amido Black 10B) or 2) the evaluation is performed in situ by spot fluorometry (Hoechst 2495). 3 The sample can be dissolved together with the acetate layer completely in dioxane, dimethylsulfoxide or N,N-dimethylformamide and evaluated photometrically or 4) fluorometrically. 5) Highest sensitivity is reached if the fluorochrome (Hoechst 2495) bound to the protein is eluted with 15% NH4OH and measured fluorometrically. There is a linear correlation with a correlation coefficient of 0.999 between the fluorescence and a protein amount of 10 ng-20 micrograms. In addition to its simplicity, the method has the advantage of being independent of or well-defined against other external influences, e.g., sodium dodecyl sulfate, mercaptoethanol, Triton X-100, etc. The stainability of a protein with Amido Black is influenced stoichiometrically by sodium dodecyl sulfate (not by mercaptoethanol) whilst the staining with Hoechst 2495 is not at all affected. As there is linear correlation between the area of a spot on an acetate layer and the volume applied in the range between 0.5 and 5 microliters, (only influenced stoichiometrically by the protein concentration in that volume, which in turn is measured by staining with Amido Black), then with a simple iterative calculation on the basis of suitable calibration curves, it is easily possible to determine a protein concentration in mg/ml even in an unknown volume between 0.5 and 5 microliters.


Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, I. Determination of the amino acid sequence of the antitryptic domain by solid-phase Edman degradation.

The acid-stable trypsin inhibitor of human serum and urine is released in vivo by limited proteolysis from the high molecular weight, acid-labile inter-alpha-trypsin inhibitor. When complexed with trypsin, both this acid-stable, active derivative and the inter-alpha-trypsin inhibitor can be degraded in vitro by prolonged digestion with trypsin to a low molecular weight "minimal" inhibitor. This minimal trypsin inhibitor was sequenced and found to be homologous to the known Kunitz-type inhibitors (e.g. the basic trypsin-kallikrein inhibitor from bovine organs). This indicates that the antitryptic activity of the big inter-alpha-trypsin inhibitor is due to a Kunitz-type domain.


Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, II. Characterization of a second inhibitory inactive domain by amino acid sequence determination.

A short digestion with excess of trypsin releases an inhibitor with an apparent molecular weight of 14,000 from both the inter-alpha-trypsin inhibitor and the ITI-related acid-stable inhibitor. The amino acid sequence of this inhibitor was determined. The inhibitor is composed of two covalently linked homologous Kunitz-type domains. One domain has antitryptic activity, as reported. This paper characterizes the second, inactive domain as also of the Kunitz type.


Kunitz-type proteinase inhibitors derived by limited proteolysis of the inter-alpha-trypsin inhibitor, III. Sequence of the two Kunitz-type domains inside the native inter-alpha-trypsin inhibitor, its biological aspects and also of its cleavage products.

The human inhibitor HI-14 consists of two Kunitz-type domains covalently connected. They are liberated from the human ITI by limited tryptic proteolysis. The inhibitor HI-14 is formed via a trypsin inhibitor complex. We have reported the amino acid sequences of the domain with antitryptic activity and the homologous domain without activity. Here we present the sequence of the domains as present in ITI. The domain lacking antitryptic activity is the N-terminal part of the inhibitor HI-14, whereas the domain with antitryptic activity represents the C-terminal part of HI-14 and probably the C-terminus of the ITI-molecule, too.


Human inter-alpha-trypsin inhibitor. Limited proteolysis by trypsin, plasmin, kallikrein and granulocytic elastase and inhibitory properties of the cleavage products.

The acid-labile inter-alpha-trypsin inhibitor is cleaved enzymatically in vivo, liberating a smaller acid-stable inhibitor. The molar ratio of native inhibitor to this smaller inhibitor in plasma is significantly changed in some severe cases of inflammation and kidney injury. To clarify this observation on a molecular basis, the action of four different types of proteinases (trypsin, plasmin, kallikrein and granulocyte elastase) on the inter-alpha-trypsin inhibitor was studied. The initial rate of cleavage of the inter-alpha-trypsin inhibitor by a 1.3-fold molar excess of proteinase over inhibitor was found to be 4375 nM x min-1 with granulocyte elastase, 860 nM x min-1 with trypsin, 67 nM x min-1 with plasmin, and 0.3 nM X min-1 with kallikrein. Obviously, of the enzymes studied so far, the granulocyte elastase known to be released during severe inflammatory processes is by far the most potent proteinase in the transformation of the inter-alpha-trypsin inhibitor. The inter-alpha-trypsin inhibitor and its cleavage products inhibit bovine trypsin very strongly (Ki = 10(-9)--10(-11) M), porcine plasmin much less strongly, human plasmin very weakly and pancreatic kallikrein practically not at all.


Demonstration of two different regions of lateral asymmetry in human Y chromosomes.

Two differently stained regions of lateral asymmetry were observed in the long arm of the human Y chromosome, following FPG staining. The first asymmetry was confined to band q12 of the long arm. The second asymmetrically stained region was located at the junction between bands q11 and q12. In the non-fluorescent Y chromosomes only one region of lateral asymmetry was found at the end of the long arm and its staining properties were similar to the region situated at the junction between q11 and q12 bands in the fluorescent Ys. The two morphologically distinguishable regions of lateral asymmetry are presumed to indicate sites containing different satellite DNAs in the human Y chromosome.


Populational polymorphisms in silver staining of nucleolus organizer regions (NORs) in human acrocentric chromosomes.

The Ag stainability of the nucleolus organizer region (NOR) was studied in the acrocentric chromosomes identified by Q banding of cultured lymphocytes in 41 karyotypically normal persons (33 males and 8 females) originating from southeast Estonia. The data obtained are compared with those established earlier for a combined Vienna-Ulm population of 51 karyotypically normal persons (see Mikelsaar et al., 1977a). Significant differences between the two populations in the frequency and patterns of Ag-positive NORs were found. The following findings were most striking: the frequency of Ag-positive NORs in chromosome 14 and in the totals was significantly lower in the Estonian population than in the Vienna-Ulm population (P less than 0.01). The average modal number of Ag-positive NORs per individual was 7.8 in the Estonian population and 8.7 in the Vienna-Ulm sample (P less than 0.01). If the data of the two populations were combined the frequency of positive NORs was significantly (P less than 0.05) lower in chromosome 22 than in 13, 15, and 21, but not 14.


Immunochemical study on basement membrane (type IV) collagens.

Basement membrane (type IV) collagens were extracted from a mouse tumour with acetic acid and from human placenta after limited enzymatic digestion. Antisera were produced against both collagens in rabbits and guinea-pigs and examined by various assays. These antisera were found to be specific for basement membrane collagen and showed little or no cross-reactions with the interstitial collagens, types I, II and III or with human placenta collagen consisting of alpha A and alpha B chains. Varying degrees of cross-reaction were observed between antisera to human and mouse type IV collagen. Immunochemical analyses demonstrated the presence of three distinct determinants in the tumour type IV collagen. Rabbit antisera against this antigen reacted with either collagenase-resistant segments or with a collagenous, disulphide-bonded segment (P3). Guinea-pig antisera recognized primarily antigenic determinants in the P3 segment. Antisera to placenta type IV collagen reacted with another collagenous, pepsin fragment (P1) which lacks disulphide bonds. These antisera showed complete cross-reaction with collagenous alpha 1 (IV) chains prepared from pepsin-digests of human placenta and bovine lens capsule.


An examination of the O and K specificity involved in the antibody-induced loss of the K88 plasmid from porcine enteropathogenic strains of Escherichia coli.

The heat-labile K88 antigen, a virulence determinant coded for by a transmissible plasmid, was eliminated from enteropathogenic strains of Escherichia coli by passage through media containing antibodies to the heat stable antigens of an Abbotstown (O149:K91,K88ac) strain. The plasmid-curing activity of O149 antisera was not O-antigen specific as O149, O45, O8 and O138 strains of E. coli could be 'cured' of their K88 plasmids by this technique. The curing activity was differentiated from the O-antibody by gel filtration, the O149 antibodies were eluted in the IgM peak while the curing activity was found in the IgG peak. In view of the lack of O-specificity and the absence of K88 antibodies it appears that antibodies to a common heat-stable antigenic determinant were involved in this phenomenon.


Idiotypic specificities and cross-reactivities of rabbit antibodies to human antidextran.

Idiotypic antibodies were prepared by immunizing two groups of rabbits with dextran-antidextran specific precipitates and purified antidextran obtained subsequently from the same human donor. Half of the animals were made tolerant to pooled human IgG. Tests showed that sera from tolerant rabbits reacted better with the antidextran preparation used to immunize the other group of animals than with the antidextran that formed part of their immunogen. Non-tolerant animals did not recognize this serological difference. Sera from animals immunized with the antidextran preparation donated later reacted better with this material irrespective of their tolerance to human IgG.


The effect of ablation of eosinophils on immediate-type hypersensitivity reactions.

The effect of ablation of eosinophils on hypersensitivity reactions in guinea-pigs was tested by administration of rabbit antiserum to the eosinophil (AES) and by administration of glucocorticoids. Both AES and methylprednisolone ablated eosinophils from the blood and peritoneal cavity of test animals. Neither administration of AES nor methylprednisolone, however, altered passive cutaneous or systemic anaphylactic reactions when compared to reactions occurring in control animals treated with normal rabbit serum (NRS). Also there was no consistent effect of AES on the intensity of the Arthus reaction. The effect of ablation of eosinophils on histamine release in the passively sensitized peritoneal cavity of the guinea-pig was also tested. In five experiments a significant reduction in histamine release was seen in AES-treated animals. Ablation of eosinophils by cortisone acetate also resulted in a marked reduction in the quantity of histamine released into the peritoneal cavity following passive sensitization and antigen challenge. Histamine release following intraperitoneal injection of compound 48/80 was not affected by either the prior administration of AES or cortisone acetate, suggesting that the stores of histamine were not depleted by these agents. Overall these results suggest that eosinophils do not play a prominent role in initial expression of immediate-type hypersensitivity reactions where the density of these cells in tissues is low. When present in larger numbers, however, eosinophils may contribute to histamine release in immediate-type reactions.


Effect of leukocyte hydrolases on bacteria. XIV. Bacteriolytic effects of human sera, synovial fluids, and purulent exudates on Staphylococcus aureus and Streptococcus faecalis: modulation by Cohn's fraction II and by polyelectrolytes.

Normal sera and plasma, derived from humans, calves, rats, rabbits, horses, human synovial fluids, inflammatory exudates, and leukocyte extracts, when sufficiently diluted are highly bacteriolytic for Staph, aureus, Strep. faecalis, B. sutilis and to a variety of gram-negative rods. On the other hand, concentrated serum or the other body fluids are usually not bacteriolytic for these bacterial species. While the lysis of Staph, aureus and B. subtilis by diluted serum is not lysozyme dependent, lysis of Strep. faecalis is absolutely dependent on the concentration of lysozyme. The lytic factor in human serum is present in Cohn's fractions III, IV, and V. It is nondialyzable, resistant to heating for 75 degrees C and 20 min, and acts optimally at pH 5.0. Like leukocyte extracts, synovial fluids, and inflammatory exudates, it lyses only young staphylococci. The inability of concentrated serum to lyse Staph. aureus and Strep. faecalis is due to the presence in the gamma globulin fraction of a potent inhibitor, which can be partly removed by dilution of by adsorption upon the homologous bacteria. Lysis of the bacteria is also strongly inhibited by Cohn's fraction II (gamma globulin) by high-molecular-weight DNA, heparin, liquoid, and histone. The possible role played by serum globulin in the protection of bacteria against degradation by leukocyte is discussed.


Effect of leukocyte hydrolases on bacteria. XV. Inhibition by antibiotics, metabolic inhibitors, and ultraviolet irradiation of the release by leukocyte extracts, trypsin, and lysozyme of lipopolysaccharide from gram-negative bacteria.

Leukocyte extracts, trypsin, and lysozyme are all capable of releasing the bulk of the LPS from S. typhi, S. typhimurium, and E. coli. Bacteria which have been killed by heat, ultraviolet irradiation, or by a variety of metabolic inhibitors and antibiotics which affect protein, DNA, RNA, and cell wall synthesis no longer yield soluble LPS following treatment with the releasing agents. On the other hand, bacteria which are resistant to certain of the antibiotics yield nearly the full amount of soluble LPS following treatment, suggesting that certain heatlabile endogenous metabolic pathways collaborate with the releasing agents in the release of LPS from the bacteria. It is suggested that some of the beneficial effects of antibiotics on infections with gram-negative bacteria may be the prevention of massive release of endotoxin by leukocyte enzymes in inflammatory sites.


Characterization of a 130,000-dalton glycoprotein isolated from pulmonary secretions of patients with alveolar proteinosis.

A new glycoprotein with an apparent molecular weight of 130,000 was isolated, purified, and partially characterized from the pulmonary secretions which accumulate so massively in the lungs of patients suffering from pulmonary alveolar proteinosis. The amino acid analysis of the glycoprotein showed the presence of relatively high amounts of glycine, glutamic acid, aspartic acid, leucine, and valine, and small amounts of hydroxyproline, but no hydroxylysine. It contains approximately 6% hexose, 3% sialic acid, and 4% glucosamine. The neutral sugars are galactose, mannose, and fucose. This alveolar glycoprotein cross-reacted with an antiserum prepared in rabbits against a larger glycoprotein (250,000 mol wt) isolated from the same source, suggesting that the larger alveolar glycoprotein may be the precursor of the smaller one.


E-rosette formation in Graves' ophthalmopathy.

We investigated the lymphocyte characteristics of 77 Graves' disease patients with and without infiltrative ophthalmopathy. Thirteen patients with infiltrative ophthalmopathy without prior antithyroid therapy and 20 euthyroid patients with progressive ophthalmopathy demonstrated decreased percentages of active and total erythrocyte rosette-forming lymphocytes compared to thyrotoxic patients without eye disease and to a control population (p less than 0.001). There was no significant difference in rosette-forming cells between untreated thyrotoxic and treated euthyroid patients with ophthalmopathy. No lymphocytotoxic antibodies or rosette inhibitory factor was present in the sera of patients with infiltrative ophthalmopathy. Untreated and treated patients with lid retraction and mild proptosis without extraocular muscle disease had decreased active rosette-forming cells (p less than 0.001) but normal total rosette-forming cells. Five patients with infiltrative ophthalmopathy who failed to improve with systemic corticosteroids demonstrated elevated active but normal total rosette-forming cells. Differences in rosette formation between ophthalmic and nonophthalmic Graves' disease may represent an associated cell-mediated abnormality that may explain why control of the thyrotoxic state need not correlate with the ophthalmic manifestations of the disorder.


Histochemical observations on Pneumocystis carinii: selective demonstration of honeycomb forms.

Histochemical investigations of pulmonary lesions indicated selective coloration of membranes of honeycomb stages of Pneumocystis carinii by the periodic acid--sodium bisulfite--resorcin-fuchsin reaction for basement membranes; mucus, fibrin and other deposits in respiratory pathways did not react. These membranes were colored selectively also by the picro-Sirius Red F3BA method for collagens; fungi in tissues from patients with candidiasis remained unstained. For simultaneous demonstration of honeycomb and cyst forms of Pneumocystis carinii, sections were prestained with Grocott's modification of Gomori's methenamine-silver nitrate technic and then treated with the periodic acid-Schiff (PAS) or picro-Sirius Red F3BA reaction. In contrast to other Gram-positive microorganisms, cysts of Pneumocystis carinii were immediately decolorized by acetone-ether mixtures; this indicates differences in the mode of dye binding. Frequently, only one stage of Pneumocystis carinii was found in a given area. Hence a combination of reactions showing different stages is recommended for studies of small tissue samples.


Quantification of nuclear DNA and intracellular glycogen in a single cell by fluorescent double-staining.

It was found that intracellular glycogen is stabilized against acid treatment when it is stored under dry conditions for three months after methanol fixation. This stabilization allowed quantitative double fluorescence staining for nuclear DNA and intracellular glycogen, in a single cell. A Feulgen nucleal reaction, with acriflavine-Schiff's reagent following 5 N HCl hydrolysis at 25 degrees C for 4 min, was followed by a pararosanilin-Schiff PAS reaction for glycogen. This short term hydrolysis was found to be sufficient for the performance of a acriflavine-Schiff's Feulgen nucleal reaction and to provide good preservation of intracellular glycogen. Quantification of nuclear DNA and intracellular glycogen were consecutively carried out with a digital microfluorometer on a single ascites cancer cell of the AH-13 line stained by this method. It was found that there is a positive linear correlation between the amount of DNA and glycogen in this cell line.