Efficacy of tri-tab (T3 uptake diagnostic kit) under extreme temperature conditions. A new surface adsorbent technique has been compared with the widely used resin sponge T3 uptake method. The new technique has the desired attributes, correlates very highly with the resin sponge method (r = 0.93) and has been accepted as a simple, rapid and reliable method under normal temperature conditions. However, under extreme conditions (0-20 degrees C and 30-60 degrees C) the test is unreliable.


[Experimental xenografting in widely divergent species: Interaction of humoral factors in hyperacute xenograft rejection in the rat-dog system (author's transl)]. The purpose of this paper is to test serum factors which are by interaction with cells responsible for the HXAR. Special stress was laid upon the examination of different serum fractions with or without the presence of preformed natural antibodies. Xenohemoperfusion: Rat kidneys were isolated and perfused in vitro with both whole blood resp. serum fractions and various perpheral cells of dogs. Rejection criterion was the diminishing renal blood flow at a constant pressure of the perfusion. Perfusate: Dog serum fractions were isolated by means of carton electrophoresis into the following fractions: Albumin, alpha-1, alpha-2, beta and gamma-globulin. Dog peripheral blood cells were separated into WBC, RBC and platelets by means of centrifugation and washing of EDTA whole blood. The serum fractions were tested for antibodies using hemagglutination and complement fixation techniques. 1. Hemoperfusion with serum fractions leads to a longer perfusion rate and to higher renal blood flow than perfusion with whole dog blood. 2. No preformed natural antibodies were found in the alpha-2 globulin fractions; however, perfusion with alpha-2 globulin fractions produced a rejection phenomenon both in combination with cells and erythrocytes. 3. Perfusion with serum fractions containing preformed natural antibodies produced a rejection only when WBC and thrombocytes were present. These results suggest that HXAR is induced by two different mechanisms: 1. The effect of humoral natural antibodies (found in the beta and gamma globulin fraction) together with peripheral cells. 2. The effect of a non-immunological humoral factor, found in the alpha-2 globulin fraction, leading to destruction of the xenogeneic organ, obviously to a minor degree dependent on interaction with peripheral cells.


Improving the specificity and yield of the contagious bovine pleuropneumonia complement fixation test antigen. Several methods for increasing yield and specificity of the contagious bovine pleuropneumonia complement fixation test antigen which is derived from Mycoplasma mycoides subsp mycoides, strain V5, were examined. Changes in culture conditions that increased the cell mass per unit volume of culture did not result in comparable increase in antigen yield. Sixteen to 60-day-old cultures yielded more boiled cell antigen than younger cultures. Some antigen in young cultures appeared to be masked, probably by galactan. The yield of antigen extracted from boiled cells with ethonol was as much for two to eight-day-old cultures as for older cultures. The ethanol extract antigen was less reactive with false positive bovine sera than standard boiled antigen while reactivity with anti Mycoplasma mycoides sera was similar to that of standard antigen. Adsorbed gamma globulin was not detected in either boiled or ethanol extract antigen. The data suggest that several complement fixing antigens were present in antigens  derived from older cultures.


[Total parotidectomy preserving the facial nerve. Significance of preoperative parotidography by staining]. The authors advise the carrying out of total partiodectomy with conservation of the facial nerve in all parotid tumors and supply a method of obtaining a truely radical parotidectomy by pre-operative vital staining.


Ground-glass hepatocytes in unselected liver biopsies. ultrastructure and relationship to hepatitis B surface antigen. 400 unselected liver biopsies were stained with aldehyde-thionin (AT) and examined for ground-glass hepatocytes (GGH). 1) AT-positive GGH were found in about half of all HBSAg-positive patients but were never seen in HBSAg-negative patients. 2) AT-positive GGH were observed in healthy HBSAg carriers and in all forms of chronic hepatitis but never in acute HBSAg-positive hepatitis. 3) AT-negative GGH were occasionally seen in HBSAg-negative patients under drug treatment. 4) Ultrastructurally GGH showed proliferation of smooth endoplasmic reticulum which only in AT-positive GGH contained filaments, 20-25 nm in diameter, within its cisternae.


IgA in commercial gamma-globulin preparations. The presence of anti-IgA has been related to adverse reactions to injection or infusion of IgA-containing material. In this study IgA was demonstrated in all of the investigated commercial gamma-globulin preparations. In the material from a few producers it was, however, very low. By immuno-gel filtration it could be shown that the IgA consisted of aggregates as well as a 7S component and fragments. By immunoelectrophoresis-immunofluorescence IgG-IgA complexes could also be detected. The IgA aggregates could be almost completely degraded by reduction-alkylation. It is implied that such changed IgA in gamma-globulin preparations for injection may increase the risk of immunization against IgA.


Antibody activity of heavy and light chains and recombined IgG of human IgG anti-D. Four human sera with high titers of IgG anti-D were studied in recombination experiments with isolated heavy and light chain. The binding to the D antigen measured by hemagglutination with O,R1R2 erythrocytes was 30-60 times weaker with isolated anti-D heavy chains and with hybrids of anti-D heavy chains and pooled light chains than with the parent anti-D IgG. Light chains and hybrids of light chains from anti-D IgG and pooled heavy chains showed no binding. This indicates that the heavy chains are more important in antigen binding than the light chains in this system. It was also shown that the idiotypes need both heavy and light chains of the anti-D molecule to give a positive reaction with specific anti-idiotypic antiserum.


The first isoallotype of human IgA proteins. An antigenic determinant occurring as allotype in the IgA2 subclass and as isotype in the IgA1 subclass. An antigenic determinant that occurs on all IgA proteins except on IgA2 proteins of the A2m2 allotype is described. The determinant is an isoallotype, being an isotype of IgA1 and at the same time an allotype of IgA2 proteins. It is proposed that the symbol nA2m2 be used for the new determinant, where n stands for isoallotype (formerly nonmarker) and A2m2 refers to the allotype to which it is antithetically related.


Coexistence of protein AA and immunoglobulin light-chain fragments in amyloid fibrils. Coexistence of amyloid fibril protein AA and homogeneous immunoglobulin light-chain fragments was found in the isolated amyloid fibrils of two patients with amyloidosis secondary to rheumatoid arthritis. The light-chain amyloid fibril protein showed antigenic identity with a light-chain amyloid from a patient with primary amyloidosis, which was identified as the VlambdaIV subgroup by amino acid sequence analysis. In the amyloid fibrils isolated from another patient with primary amyloidosis there was a mixture of VlambdaIV and VlambdaV homogeneous immunoglobulin light chains. Thus, a mixture of protein AA had lambda light chains or two different types of homogeneous light chains may be found in the amyloid fibrils of some patients.


Human agglutinins to rabbit erythrocytes. Characterization of an antigen. Crude preparations inhibiting the agglutination of rabbit erythrocytes by pooled human serum were obtained by extraction of erythrocyte stromata with various solvents (chloroform/methanol, trichloroacetic acid, and so forth). Highest inhibiting activity was found in a preparation containing sphingoglycolipid; 0.8 mug inhibited 2 agglutinating units of serum. The active preparations retained their activity after 5 min of incubation in a boiling water bath and after treatment with proteolytic enzymes or neuraminidase. Treatment with periodate abolished the activity. The antigenic site is therefore probably determined by the sugars of the sphingoglycolipid.


Cytological changes in the urine in the form of inclusion-bearing cells, giant cells and haematuria after vaccination with inactivated influenza virus vaccine. A study with application of Millipore procedure and Papanicolaou staining. Marked cellular reactions in the urine appeared after subcutaneous vaccination with inactivated influenza virus vaccine. Inclusion-bearing epithelial cells increased in 16 cases from 2.4 to 9.6% and erythrocytes from 2.5 to 16.2 per visual field. Pathological values for inclusion-bearing cells were found in 5 of 18 cases (28%), spherical giant cells in 3 (17%), and an abnormal number of erythrocytes in 7 (39%). Killed virus, lacking the capability of replication, thus gave rise both to the formation of inclusions and to fusion of cells within the epithelium of the kidneys and of the urinary tract, and also to haematuria. There was a striking resemblance to the reaction in 34 cases of uncomplicated influenza A2, but the inclusion-provoking effect of the vaccination dose was clearly weaker and the erythrocyturia somewhat less pronounced. As a rule the cellular changes after the vaccination quickly reverted to normal, but there were examples of elevated values still existing after 6 months. Two patients had a high inclusion content and haematuria persisting for 3 and 4 years, one of whom following influenza A2. Influenza and influenza vaccinations may be the cause of certain instances of chronic benign haematuria.


Cephazolin sodium in the management of complicated urinary tract infection. Forty-three patients were treated with cephazolin sodium, a parenteral cephalosporin antibiotic for 45 episodes of urinary tract infection complicated by a variety of underlying conditions. In 42 episodes, there was a satisfactory clinical response, and in 37 episodes this was associated with elimination of the bacterial pathogen from the urine. In 21 out of 31 patients available for examination 3 months later, the urine was still free of bacteria. A relatively prolonged plasma half-life and high urinary concentrations of the drug permit successful treatment of urinary tract infections with injections given only twice daily.


Serologic differences in strains of Sporothrix schenckii. To obtain evidence that Sporothrix scheneckii enters the body by contact with contaminated materials, the antigenic property of strains from different sources was investigated. The reciprocal absorption test of the antisera against a soil isolate and a human isolate (KO 4606) showed that the absorbed antisera against KO 4606 possessed unique antigen(s) in addition to the common antigen of both strains. Twenty-three clinical isolates were tested with absorbed antisera. Not all of them possessed the unique antigen(s), but there were serologic varieties among S. schenckii strains, regardless of their sources, clinical type of the disease and the morphology of the yeast phase cells.


Electron microscopic, histochemical and disc gel electrophoretic studies on the deoxycholate soluble proteins from human plantar horny layers. The matrix proteins of human plantar horny layers were extracted with deoxycholate, purified and examined with the light and electron microscopes, and their histochemical characteristics were investigated. About 44.3% of the total proteins was released into the 270,000 X g supernatant fraction, following incubation with deoxycholate, and the precipitate obtained by dialysing this supernatant fraction consisted of fine granular materials which were found to be electron dense, round material under the electronmicroscopy. Small electron dense dots were seen on the surface of this material. This pellet was positive with Pauly's reagent, toluidine blue and Harris hematoxylin-eosin. However, the material further purified by the molecular sieve chromatography was negative with Pauly's reagent. And it had been proved to be of keratohyalin granules origin by the indirect immunofluorescent study. Another protein fraction which was obtained by the molecular sieve chromatography was positive with diazotized sulfanilic acid and showed tinctorial properties as keratohyalin granules show in vivo.


The metabolic fate of chlormadinone acetate in the baboon. The metabolic fate of chlormadinone acetate (17alpha-acetoxy-6-chloro-4, 6-pregnadiene-3, 20-dione; CAP) was studied in intact and biliary fistula baboons. The steroid was labeled with 3H at position 1 and with 14C at the carboxyl moiety of the 17alpha-acetate, thus affording the opportunity to ascertain the loss of the 17alpha-acetoxy group and the fate of both labels. The averages of the radioactivity excreted, given as percentages of the amounts injected, and the standard deviations were as follows: In the urine of intact animals after 6 hours, 5.7 +/- 0.2% and 5.5 +/- 0.7% of the 3H and 14C were recovered, respectively. After 6 days, there was 17.5% of the 3H and 16.2% of the 14C in the urine plus 15.3% of the 3H and 16.4% of the 14C in the feces. In baboons with biliary fistulas, the total radioactivity excreted was 7.8 +/- 0.7% of the 3H and 11.6% of the 14C in the urine, and 30.9 +/- 4.4% of the 3H and 30.7% of the 14C in the bile after 6 hours. Glucosiduronates were the predominant conjugates in the urine and bile. The similarity in the urinary excretion of radioactivity in the first 6 hours in intact and biliary fistula animals, the relatively low excretion of radioactivity in the bile and after 6 days in the urine, and the low fecal excretion suggest that the metabolites of CAP are not involved in an extensive enterohepatic circulation in the baboon. Deacetylation of the 17alpha-acetate in CAP was detected in the early collection periods of the urine and bile and constituted a very small percentage of the injected compound. No significant oxygenation of CAP at position 1 was detected. The metabolism of CAP is discussed and compared to our previously reported data on the metabolism of progesterone, ethynodiol diacetate and medroxyprogesterone acetate and the data on other progestogens reported in the literature. It appears that the excretion of CAP is significantly slower in the baboon than that of the other progestogens. The amounts of glucosiduronates of CAP and/or its metabolites formed in vivo are less than those formed with the other progestogens. Also, the extent of deacetylation of the 17alpha-acetate of CAP is much less than that of the 3beta-acetate of ethynodiol diacetate.


[A method of submacroscopic study of the whole human breast (author's transl)]. A method of submacroscopic study of the whole human breast is described. Breasts are removed surgically or at autopsy. Serial thin sections (2mm.) are defatted, stained, clarified, preserved in sealed plastic bags, observed under a dissecting microscope and photographed. The lesions found are interpreted and removed for histological examination. The method is simple, allows a careful and complete study of the glandular lesions and appears to be suitable for studies on origin and development of human breast cancer.


New look at pelvic exenteration.

New look at pelvic exenteration. Our experience with 18 patients undergoing pelvic exenteration for advanced primary or recurrent pelvic malignancies is presented. Only one postoperative death was noted, and morbidity was minimal despite the advanced age and high incidence of radiotherapy failures seen in our patients. Although no improvement in cure of malignancy has been seen in this small series, appreciable periods of symptom-free life have been achieved in patients who were previously incapacitated by extensive pelvic pain, fistulas, sepsis, hemorrhage and urinary-fecal incontinence. Because of the symptomatic palliation obtained in our experience, with minimal morbidity and mortality, we have developed a liberal attitude toward the use of pelvic exenteration in the management of selected patients with extensive pelvic malignancy, even when cure is not anticipated.


[Lipodystrophia intestinalis (Whipple's disease)--a rare origin of a sprue syndrome]. An own case of lipodystrophia intestinalis (Whipple's disease) of a male patient who was 41 years old at the time of the first admission is reported and discussed using literature. The biopsy of the small intestine is of decisive importance for the establishment of the diagnosis as well as for the long-term therapy with antibiotics (1 g oxytetracycline a day).


[Hazards of hyperthyroidism therapy].

[Hazards of hyperthyroidism therapy]. It is reported on the competitive methods of treatment of hyperthyroidism and their side-effects, whereby special value is laid to the choice of the method, the consideration of late complications and the observation of the therapy. With the help of casuistic instances complications such as postoperative exacerbation of hyperthyroidism, postoperative hypothyroidism as well as the sequelae of irrelevant dosage of the antithyroidal medicaments are explained.


[Reflex-dystrophic pseudo-arthritis or Sudeck syndrome (author's transl) ]. Local bone- and tissue dystrophies identical with Sudeck's syndrome and progressing in 3 stages are called reflex dystrophies. In the 1st and 2nd stages complete recovery is possible, in the 3rd stage only irreversible dysptrophic-ankylosing defect-healing. Etiologic significance of causal factors discussed is not certain. In, initially similar, inflammatory and rheumatic arthritis often only the further course and radiologic appearance provide a diagnosis. So far we have no desirable clear definition of reflex dystrophic conditions.


[Sub-ppm determination of thiouraciles applicated as thyreostatics in meat by high pressure liquid chromatography (author's transl)]. The thyreostatics 2-thiouracile, 4-methyl-2-thiouracile, and 2-propyl-2-thiouracile were extracted from meat and determined by High Pressure Liquid Chromatography to a limit of about one ppb.


[Resection syndrome following transurethral resection of the prostate]. The essence and clinical aspects of the resection syndrome occurring in association with transuretheral prostate resection and the possiblity of determining the direction of the absorption of the washing solution are discussed. By means of radioisotope tests a direct correlation was detected between the extent of the haemorrhage and the intravasation of the washing solution. Great importance is ascribed to the vesico-renal reflux often occurring during resection, which might have a major role in the development of post-resectional pyelonephritis.


[Studies on the genesis of so-called Sjögren cells in supravital preparations]. 1. The effect of the LE-cell factor lies in nuclear blockade; the normal degenerative nuclear swelling and lysis is inhibited. 2. The time of the nuclear blockade and thus the appearance of the LE-bodies depend on the activity of the LE-serum. 3. High-titred sera lead to an early nuclear blockade. Small, rigid, dark, clump-like LE-bodies appear rapidly and are fully phagocytosed. 4. Our experiments have shown that with decreasing activity of the LE-sera the nuclear blockade appears later and more slowly. Such LE-bodies are relatively large and homogenous and have lost some rigidity. They are correspondingly late and only partly phagocytosed. So-called granulocytic and monocytic Sjögren-cells appear. 5. We found that quantitative differences of the causal factors lead on the one hand to LE-cell formation, on the other to the partial phagocytosis phenomenon. Therefore, we prefer not to label the Sjögren-cells as pseudo-LE-cells. 6. It has again been shown that the standardized supravital preparation is not only a simple method for demonstrating LE-cells, but is also superior to other methods used because of its greater specifity and sensitivity. Besides the judgement of cell vitality, many hours after making the preparation phagocytosis can be observed.


[Palliative treatment of nonresectable cardia tumors using esophago-jejunal bypass]. Transdiaphragmal internal bypass with Roux's loop is believed to be the best solution for non-resectable primary or secondary tumorous obstructions in the region of the cardia. Of 26 cases subjected to such an operation, 3 patients died. The indication of bypass operations must not be extended at the cost of palliative (and even less of radical) resections. On the contrary, in the surgery of gastric tumours these operations aim to improve operability. It is believed that internal palliative solutions might finally replace the external ones, since the former intervention offers definite nutritional and moral advantages.


Mortality after laparotomy. A 10-year series. The Department of Surgery at the University Hospital, Linköping has 133 beds and serves a population of 130000. During the 10-year period 1962-1971 35039 in-patient operations were performed, of which 16719 (48%) were laparotomies. If fracture surgery and urological operations are excluded (during the relevant period these specialties were included under General Surgery) the proportion of laparotomies becomes 67%. Of patients subjected to this procedure 431 (2.58%) died. The mortality for appendicectomy was 0.2%, for gall-bladder surgery 1.3%, for gastric surgery 8.1%, for colon surgery 11.9% for small-gut surgery (including ileus) 15.2%, and for pancreatic surgery 22.2%. Over and above the target organ, malignancy and age exceeding 50 years appear to have contributed greatly to a lethal outcome.


Unexpected findings with the new chromosome banding techniques in a patient formerly diagnosed as having G-deletion syndrome II. A girl is described, who, upon her first admission to our pediatric department in 1965, was supposed to have G-deletion syndrome II on the base of the chromosome findings in routine orcein-stained preparations and her clinical aspect. When, however, in 1972 a Q- and G-banding analysis was performed, the patient seemed to be a t(14q+; 22q-) translocation carrier. These findings are discussed in relation to the known heterogeneity of the clinical picture of patients previously reported as having a G-deletion syndrome and in whom no banding studies were done. Finally, the need for more extensive studies with the different techniques is stressed, particularly, in cases of Gq- chromosomes, of which the familial occurrence is suggestive of a hidden reciprocal translocation.


Leucocyte morphology and chromosome morphology. The banding techniques currently employed in human cytogenetics for the identification of the individual chromosomes have been used to stain PHA lymphocytes and circulating leucocytes. The capacity of these techniques to localize singular chromosomes or chromosomal regions has been investigated. It has been observed that among the four major categories of bands (Q, G, E and R) only the quinacrine staining is informative in interphase nuclei, because of its peculiarity to stain the long arm of the Y chromosome and few other heterochromatic regions. Interphase nuclei treated according to the C-bands show the presence of several heterochromatic masses, corresponding to the centromeric areas of individual chromosomes, but as such they cannot be recognized accurately. More specific and selective techniques, like G-11 and G-Y protocols, appear to be suitable to localize the centromeric regions of chromosome no. 9 and and long arm of Y chromosome. Variation of the incubation time in the alkali-saline solutions and of pH values have proven to be appropriate for the demonstration of other heterochromatic regions in interphase nuclei and in circulating leucocytes. The "nuclear" approach to study of specific heterochromatic regions of human chromosomes may be of practical interest into the investigation of several biological problems and into the detection of individuals carrying chromosome variants.


Enzymatic destruction of the elastic lamella at the mouth of cerebral berry aneurysm? An ultrastructural study with special regard to the elastic tissue. Destruction of the elastic tissue is probably an acquired lesion and is decisive for the appearance of cerebral arterial aneurysms at the sites of congenital media defects. The elastic component in the mouths of aneurysms has therefore been studied by electron microscopy, using two new staining methods, i.e. ruthenium-red staining and prolonged osmium-tetroxide treatment. The hypertrophic, duplicated, elastic lamellae showed a disintegration of their luminal portions not earlier described. In close connection with the disintegrated portions, extracellular lysosome-like granules were observed. It is hypothesized that discharged leucocyte grannules containing elastase help to destroy the elastic lamellae.


A long-term comparative study of the mammalian larynx, based on whole organ serial sectioning. The paper includes brief details of a long term three-dimensional study into the fine structure of the mammalian larynx using whole organ serial sections. Differential staining, including Glees silver technique, is essential if maximum information is to be gained. A specially designed punch card will enable any future investigator to rapidly retrieve relevant information and it is expected that specimens of all available mammalian larynges will be included. Illustrations are included in this paper of the cat, dog, horse and mongoose.


The cochlear nuclei in monkeys after dihydrostreptomycin or noise exposure. The cochlea and the cochlear nuclei in monkeys subjected to dihydrostreptomycin treatment or loud noise are investigated. Their cell populations are compared with the findings in the normal monkey.


Demonstration of two distinct macrophage specific antigenic determinants in rats. Exhaustive cross-absorptions of rabbit anti-rat alveolar macrophage sera with peritoneal macrophages and of anti-rat peritoneal macrophage sera with alveolar macrophages did not prove the existence of antigenic determinants specific for alveolar or peritoneal macrophages. Titer-differences of these sera on RAM and RPM seem to be caused by a different distribution of two or more macrophage specific determinants present in both cell populations. Macrophages from other rat tissues show reactivity with these two sera different as well from alveolar as from peritoneal macrophages.


Enhanced in vitro immunogenicity of H-2 antigens in the presence of la antigens. In a primary and secondary MLC the in vitro immunogenicity of allogeneic PHA induced blast cells (which lack i region coded determinants) was compared to that of LPS imduced blast cells. Unlike LPS induced blast lymphocytes (stimulator cells) which induced high cytotoxic activity, PHA induced blast cells were found to be poor stimulator cells in a primary MLC. Yet in a secondary MLC both types of stimulator cells induced cytotoxic activity equally well. Using one type of responder cells the relative immunogenicity of various stimulator  cells incompatible on either the H-2K and H-2D region or on the i region, or on the complete H-2 complex, was compared. The magnitude of cytotoxic response induced in a strain combination differing at the complete H-2 complex exceeded by far the sum of separate responses obtained against the H-2D region, H-2K region and the i region coded determinants respectively. These results suggest that the  presence of i region coded determinants on allogeneic stimulator cells enhance the in vitro immunogenicity of H-2K and H-2D region coded transplanatation antigens.


Macrophage killing capacity. Aspects of mechanism. Immune macrophages have the capacity to kill in mixed macrophage cultures (MMC) allogeneic macrophages bearing the alloantigens used for immunization. The killing is expressed by a lytic mechanism as neither phagocytosis nor fusion between effector and target cells could be detected by E.M. T but not B lymphocytes isolated from immune spleen cells, are able to "arm" syngeneic nonimmune macrophages and render them cytotoxic. It is suggested that mediator(s) and/or membranal components are released from T cells and attach to macrophages. Such an "arming" factor enables the macrophage to recognize and kill target cells.


The pathophysiology of sleep disorders in pediatrics. Part II. Sleep disorders in children. In this part of the chapter we have used new terminology and developed a new system for classification of sleep disorders in children. We suggest that excessive daytime sleepiness should be investigated by clinicians before troubles at school necessitate referral. The narcolepsy-hypersomnia syndrome generally has not been recognized in the pediatric age group. Symptoms of excessive fear of falling asleep need to be viewed in this context. Sleep apnea-hypersomnia has received insufficient attention in the American literature. It is a syndrome that affects both adults and children with potentially disastrous cardiovascular and pulmonary complications. The relationship of the sleep apnea-hypersomnia syndrome to the sudded infant death syndrome remains speculative, although preliminary results from our longitudinal study have indicated a possible link. Both the narcolepsy-hypersomnia and the sleep apnea-hypersomnia syndromes are reviewed in detail. In contrast, we review briefly the NREM dyssomnias, including night terrors, sleepwalking, sleep talking and enuresis. All are well known to clinicians dealing with children, and we have related them to findings emanating from the sleep laboratory. We suggest that they are physiologically rather than psychogenically based and frequently represent immaturities of the central nervous system. Finally, the insomnias of childhood are presented. We emphasize that they are rare, and after ruling out organic conditions and drug-dependency syndromes, cultural styles or family stresses generally account for the majority of complaints.


Malignant lymphomas of the salivary glands. Review of the literature and report of 33 new cases, including four cases associated with the lymphoepithelial lesion. Malignant lymphomas involving major salivary glands have been reported to occur in 31 cases. To these, we add 33 cases, 17 of which were studied in detail from the points of view of clinical presentation, classification (Rappaport), staging (Ann Arbor), therapy, and subsequent course. The parotid gland was involved much more frequently than the submandibular gland. No example of sublingual gland involvement was discovered in the present series. Most of the patients were in the sixth and seventh decades of life. Noteworthy is that all sub-types of lymphosarcoma were encountered, with only a single case of Hodgkin's disease present. In four of the patients, the characteristic histologic picture of lymphoepithelial lesion was found in juxtaposition to malignant lymphoma. The majority of patients received one or more courses of postoperative radiotherapy, which offered the best chance for long-term remission. However, only 40% of patients were free of disease after two years.


Acute monocytic leukemia. Cytologic, histologic, cytochemical, ultrastructural, and cytogenetic observations. A case of acute monocytic leukemia (AMoL) studied by cytologic, histologic, cytochemical, ultrastructural, and cytogenetic technics is reported. The immature monocytes were characterized by strong fluoride-sensitive nonspecific esterase activity. Nuclear irregularity, prominent nucleoli, erythrophagocytosis, and pinocytosis were observed on transmission electron microscopy, whereas broad-based ruffles, similar to those found on normal monocytes, were identified with scanning electron microscopy. Cytogenetic analysis showed an 8/9 translocation and four or five identical 8p- marker chromosomes in most bone marrow cells.


An immunoperoxidase technic for the demonstration of the hepatitis B surface antigen in human livers. A sensitive method for demonstrating the site of hepatitis B surface antigen (HBsAg) in fixed tissues embedded in either paraffin or araldite is described. The method employs the peroxidase-rabbit antiperoxidase linkage through goat antirabbit to rabbit anti-HBsAg. In staining hepatitis antigen in agar, comparison of fixation (using three common fixatives) with unfixed precipitation arcs revealed no recognizable differences in antigenicity induced by fixation. The method allows confirmation of positive reaction by appropriate blocking controls. The technic is compared with the orcein stain of Shikata and found to be somewhat more sensitive but slightly more time-consuming.


Alpha1-antitrypsin and alpha-fetoprotein. Protein markers in endodermal sinus (yolk sac) tumors. A combined immunocytochemical and quantitative serum and tissue study was performed on a group of endodermal sinus (yolk sac) tumors, localizing and measuring both alpha1-antitrypsin (AAT) and alpha-fetoprotein (AFP) in tumor tissue and patient sera. Utilizing indirect immunofluorescent and triple-sandwich immunoperoxidase methods, both proteins were demonstrated within intra- and extracellular periodic acid-Schiff-positive hyaline globules characteristic of the tumor, as well as within the cytoplasm of tumor epithelial cells lining endodermal sinuses, where AAT deposition predominated. Tumor tissue extracts confirmed the presence of significant quantities of both proteins, and pretreatment serum elevations of both showed a parallel decline during therapy. In this study, AAT is characterized as a tumor protein marker for the first time, and a parallelism between AAT and AFP is demonstrated in both serum and tumor tissue. These findings represent additional supportive evidence for the yolk sac origin of endodermal sinus tumors in man.


Human serum dopamine beta-hydroxylase: correlation of enzymatic activity with immunoreactive protein in genetically defined samples. An antibody against human adrenal dopamine beta-hydroxylase (DBH) was used to quantitate immunoreactive DBH protein in human serum by an immunoprecipitation technique. A significant correlation was found between DBH enzyme activity and immunoreactive DBH protein in randomly selected serum samples (r = 0.94; N = 38; p less than .001). Studies of sera from obligate heterozygotes and individuals homozygous for the allele responsible for very low serum DBH enzymatic activity were compatible with a genetically mediated decrease in the quantity of circulating DBH protein in these subjects.


Effects of amyloid induction on plasma protein turnover, and its implication. Using polyacrylamide gel electrophoresis and radioiodinated plasma proteins, data on the rate constant for synthesis and fractional degradation rates for albumin, alpha1-, alpha2-, beta-, and gamma-globulins were obtained during accelerated amyloid induction in a murine model. The results indicate that during amyloid induction there is an increased rate of synthesis of alpha2-, beta-, and gamma-globulins but only the alpha2-globulin degradation rate is accelerated. In this experimental system, should amyloid protein be a degradation product of a plasma fraction, the alpha2-globulin appears as the most likely precursor. The implications of our findings are discussed, and a new general mechanism of amyloid production is proposed.


Pelvic exenteration as palliation of malignant disease. It has been traditional to exclude patients with radiation-recurrent carcinoma of the uterine cervix or other pelvic neoplasms, incapacitating pelvic pain, postirradiation fistulas, hemorrhage, or malodorous draining tumor necrosis from pelvic exenteration if cure of the malignant disease is not achievable. This negative attitude is a direct result of the reported high morbidity, prohibitive mortality, and low salvage rate previously associated with pelvic exenteration, the only acceptable surgical approach to these diseases. A recent experience with eighteen patients who underwent pelvic exenteration for advanced primary or recurrent carcinoma of the cervix, urinary bladder, or rectum has led us to challenge several traditional concepts regarding this operative procedure. We have observed but one operative death and our morbidity has been minimal. This may reflect our belief that an aggressive pelvic lymphadenectomy in those patients with direct visceral involvement from radiation-recurrent carcinoma of the pelvic viscera is not advantageous since no significant survival has ever been documented for patients with pathologic visceral involvement and positive lymph nodes. In addition, significant morbidity has always been associated directly with pelvic lymphadenectomy in the irradiated pelvis, and elimination of this phase of the operation in selected patients with radiation-recurrent carcinoma is indicated. Moreover, the considerable decrease in morbidity and the minimal mortality observed have led us to adopt a very liberal attitude toward preoperative selection criteria, and we regularly now use pelvic exenteration not only for cure but as intentional palliation in selected patients. We strongly believe that elimination of pain, fistulas, pelvic sepsis, hemorrhage, and malodorous areas of tumor necrosis are important for improving the quality of life for both the patient and family.


Cancer of the pancreas. Palliative operation, Whipple procedure, or total pancreatectomy? The choice of operation for ductal carcinoma of the pancreas is as yet not clear. Failure to make an early diagnosis still stands out as the major problem in the treatment of this disease and as a result the resectability rate is exceedingly low. Between 1964 and 1973, nineteen total pancreatectomies were performed at Peter Bent Brigham Hospital for all tumors of the pancreas. Sixteen of these were for ductal carcinoma. The mortality was 12.5 per cent. When total pancreatectomy is compared with the Whipple procedure and a simple bypass procedure and when the tumor disease encountered is corrected for stage of disease, it is apparent that total pancreatectomy carries a statistically significant longer survival for patients with Stages I and II disease (no lymph node involvement) than the other two procedures. For Stages III and IV, there is no difference in survival between the three different operations. We therefore conclude that total pancreatectomy carries a better survival prognosis than other procedures performed for Stage I and II ductal carcinoma of the pancreas.


A serum factor in aspirin intolerance disease. Direct and indirect intradermal serum challenge with aspirin intolerant sera in both aspirin tolerant and intolerant patients demonstrated a heat, cold and storage stable serum factor capable of inducing acute anti-hemostatic effects and in some aspirin intolerant patients exacerbation of brief induced clinical symptoms. The data supports a hypothesis for possible induced, conformationally altered gamma globulin resulting from low molecular weight chemical exposures such as aspirin with ensuing recognition by a genetically determined subpopulation of B-lymphocytes with surface membrane receptors avid for the Fc portion of altered gamma globulin or some conformationally altered gamma globulin fraction leading to immune and/or interprotein complexes with subsequent alterations of adrenergic, cholinergic and/or metabolic pharmacologic pathways.


Senile ectropion and entropion: a comparative histopathological study. A microscopic histopathological study was done on 500 full-eyelid-thickness surgical specimens: 25 with the diagnosis of senile ectropion and 25 with that of senile entropion. Five different staining techniques were used. There appears to be significantly more orbicularis and Riolan's muscle ischemia, atrophy, and collagen fragmentation with ectropion than with entropion. Entropion shows more septal and tarsal atrophy. In both conditions, the skin and conjunctiva show chronic inflammation and scarring as a constant feature. Statistical significance at the 1% level was present for all six characteristics studied. These histopathological changes, if not etiological, are at least concomitant features differentiating senile ectropion from entropion at the microscopic tissue level.


Blood substitutes.

Blood substitutes. With the development of modern methods of surgery, anaesthesia, and pre- and postoperative care the requirement for blood substitutes is continuously increasing. We present a review of the different blood substitutes which are already in clinical use or in an advanced stage of experimental investigation for possible practical administration. Our own clinical experience with dextrans and experimental studies on stroma-free haemoglobin and hydroxyethyl starch solutions are described.


Observations on the automated calculation of radioimmunoassay results. Radioimmunoassay standard curves derived by four different automated calculation methods are compared with those derived manually for five assays of clinical importance (thyroxine, triiodothyronine, thyrotrophin, alphafetoprotein, and human growth hormone). The three curve fitting methods (linear or cubic regression on log x versus logit y, cubic regression on log x versus y) produced considerable distortion of the manually derived curves, which in some cases could have impaired the accuracy of an estimate in a clinical sample and altered the clinical interpretation. Distortion patterns varied with the particular antigen assayed and with experimental conditions. In contrast, a simple linear interpolation method produced little distortion and gave results which were close to those derived manually in each assay examined.


Production, isolation, and properties of azetomycins. Streptomyces antibioticus synthesizes five actinomycins that differ in the "proline site" of the molecule. When cultured in the presence of azetidine-2-carboxylic acid (AzC), antibiotic synthesis was stimulated 40 to 50%, synthesis of actinomycin IV was inhibited, and one or both prolines were replaced by AzC. AzC incorporation could not be reversed by concomitant supplementation with proline or sarcosine, and only pipecolic acid affected a minor reversal of AzC incorporation. AzC-containing actinomycins were isolated and designated azet-I and azet-II; a third unresolved component or mixture was called azet-III. The molar ratio of AzC to proline was: azet-I, 1:1; azet-II, 2:0. Azet-III was equivocal. These azetidine actinomycins (azetomycins) were found to be potently inhibitory to the growth of selected gram-positive but not as potent to the growth of gram-negative organisms. The relative inhibitory affect against growth and ribonucleic acid synthesis in Bacillus subtilis was: actinomycin IV =/> azet-I > azet-II >>> azet-III. Protein synthesis was affected similarly; however, kinetic studies with B. subtilis revealed that ribonucleic acid synthesis was inhibited rapidly followed by an inhibition of protein synthesis. At concentrations less than 1 mug/ml, deoxyribonucleic acid synthesis was stimulated by these actinomycins.


Biogenetic origin of the D-isoleucine and N-methyl-L-alloisoleucine residues in the actinomycins. Studies with (14)C-labeled isoleucine stereisomers have established that l-alloisoleucine, d-alloisoleucine, and d-isoleucine may function as precursors for the biogenesis of d-isoleucine and N-methyl-l-alloisoleucine residues in actinomycin. l-[(14)C]isoleucine appears to be employed chiefly for d-alloisoleucine (and N-methylisoleucine [?] formation); however, its role in the biosynthesis of d-isoleucine and N-methylalloisoleucine remains unclear. The potential pathway of biosynthesis of d-isoleucine and N-methyl-l-isoleucine is discussed.


Effect of bleomycin on deoxyribonucleic acid synthesis in toluene-treated Escherichia coli cells. The antibiotic bleomycin stimulates deoxyribonucleic acid (DNA) synthesis in toluene-treated Escherichia coli cells. The increase in synthesis is linear with bleomycin concentration. Bleomycin-stimulated DNA synthesis is independent of replication and dependent on DNA polymerase I. Replication is spared as the DNA polymerase I-dependent DNA synthesis increases. Bleomycin does not appear to have any effect on purified E. coli DNA polymerases I or II. Our results suggest that bleomycin causes nicking of the bacterial chromosome with subsequent DNA synthesis catalyzed by DNA polymerase I.


A study of the antigenic relationships of isolates of Trypanosoma brucei from three areas in East Africa. Eleven stabilates of T. brucei, prepared from isolates collected from cattle and tsetse flies in three areas of East Africa, were compared serologically by direct agglutination tests, using (a) stabilate-specific antisera prepared in rats and (b)antisera to the predominant variant antigens of each isolate prepared in rabbits. The results indicated that groups of isolates from any one area tended to produce antigens in common, whereas isolates from different areas were generally antigenically distinct. The comparison of isolates of T. brucei on the basis of their predominant variant antigens, as a technique, was more simple to effect, and more sensitive in the detection of antigenic relationships, than comparison founded on basic strain antigens.


Mustard procedure after Baffes operation. Patients in whom Baffes' procedure has been done for palliative treatment of transposition of the great arteries may develop symptom recurrence later in life that deserves further treatment. A modified Mustard procedure is suitable for this purpose; the main difficulty in performing a formal Mustard operation is the proximity of the superior vena cava and of the inferior vena caval graft openings that enable the left pulmonary veins to drain without obstruction into the new atrium. This report deals with a 15-year-old patient in whom a modified Mustard technique was employed as a palliative method.


Spectrophotometric assays for warfarin sodium and dicumarol. Use in rapid detection of patients suspected of drug intoxication or surreptitious noningestion. The usefulness of assays for the rapid identification and determination of quantitative plasma levels of warfarin sodium and dicumarol is documented by the case histories of five patients: a man who accidentally took dicumarol for several weeks and developed an acute condition within the abdomen, a man who ingested 500 mg of warfarin sodium in a suicide attempt, a malingering nurse who surreptitiously took dicumarol, a nurse with warfarin intoxication who did not follow dosage prescription because of fear of developing thrombosis, and a woman with calf vein thrombosis who did not ingest the administered warfin sodium becausing of fear of developing bleeding. In every patient, the diagnosis of surreptitious ingestion or noningestion was suspected on the basis of the coagulation profile pattern. Confirmation of the disorder by the spectrophotometric assay enable the clinician to choose the appropriate course of action with speed and purpose.


Effects of hypothalamic lesions on levels of plasma free fatty acids in the mallard duck. Plasma free fatty acid (FFA) levels were measured in the mallard duck (Anas platyrhynchos) following hypothalamic lesions at various sites. The results indicate that ventromedial lesions produced hyperphagia,  increased deposition of fat, and significantly elevated levels of plasma FFA. Anterior bilateral lesions resulted in aphagia, severe loss in body weight and a marked decrease in plasma FFA. Lesions in other regions of the hypothalamus produced various changes depending upon the extent of damage. The neural and neuroendocrine mechanisms which regulate FFA levels in the blood are discussed with respect to the involvement of pituitary hormones.


Effect of gonadotropins and prostaglandin on 3' : 5'-cyclic AMP synthesis in young rat testis. Formation of cAMP has been studied in neonatal rat testis. In the presence of aminoophylline, LH and PGE1 stimulated cAMP levels during the entire postnatal period. The highest stimulation was found in the testis of 2-day-old rats after-which cAMP synthesis diminished with advancing age. FSH had no stimulatory effect on cAMP level in 5-day-old rats.


[Effect of ambient temperature on metabolism and heart rate of resting albino rats]. Heart rate and oxygen consumption were measured simultaneously in albino rats. These measurements were carried out in the resting animal at different temperatures between 18 degrees and 33 degrees C. The metabolism evolution with the environmental temperature allowed to place the thermal neutrality at 29 degrees C. The resting heart rate varies as metabolism. It shows the lowest values round the thermoneutrality and increases quickly as the environment is cooling. This result shows the effect of the thermal environment upon the resting heart rate level. On the other hand, the non linear relationship between metabolism and heart rate points out that the heart rate increase is not the only factor that allows an increased oxygen consumption during the body temperature regulation.


[Sensitization of bradycardia during final hypotension induced by serotonin in rats: effect of lidocaine]. It was studied the sensibilizing effect of lidocaine (8.5 mg/kg, i.v.) on the ECG (heart rate, P-R interval, QRS complex and Q-T interval) of both intact and bilaterally vagotomised rats, in the nadir of the final hypotension determined by serotonine (60  mug/kg, i.v.). The results showed (1) a certain degree of selectivity of the vagi, and (2) the effects of serotonine either isolated or associated to lidocaine on P-R interval and heart rate were reinforced when intact animals were used. Although no significant alterations of Q-T were elicited by the drugs, lidocaine surprisingly enlarged the QRS complex in a more significant fashion for the intact than for the vagotomised animals.


Glomerular metabolic changes in compensatory renal hypertrophy. In compensatory hypertrophy, 7 days after unilateral nephrectomy, an increase in respiration and glucose-6-P dehydrogenase activity are observed in the glomeruli of the hypertrophied kidney. It is suggested that is is related to the clearing of an increased amount of filtration residues by the glomerular cells.


[Radioimmunoassay of testosterone in late fetal and newborn rabbit plasma, gonads and adrenal glands]. A radioimmunoassay was used for measuring testosterone in the plasma, gonads and adrenals of 28, 29, 30 and 31-day-old rabbit fetuses of both sexes and newborns. A marked sex difference was shown in the concentrations of testosterone in plasma and in gonads whereas in adrenals the levels of testosterone were low in both sexes (34 to 147 pg/10 mg). In male fetuses, plasma testosterone levels increased from the 28th (133 +/- 20 pg/ml) to the 31st day (361 +/- 119 pg/ml) of intrauterine life, reaching then the values observed in the newborns (387 +/- 73 pg/ml). Plasma from males, on the other hand contained, at all stages studied, significantly more testosterone than plasma from female fetuses (21 +/- 6 to 41 +/- 11 pg/ml) and female newborns (42 +/- 6 pg/ml). In the same way, fetal testicular testosterone concentrations varying from 1 382 +/- 218 to 2 317 +/- 333 pg/10 mg were similar to those measured in the newborns (1 940 +/- 304 pg/10 mg) and significantly higher than fetal (13 to 34 pg/10 mg) or neonatal (44 pg/10 mg) ovarian concentrations. These results showed at evidence the endocrine activity of the fetal testis during this period.


[Polymorphism of proteins in 3 strains of domestic quail (Coturnix coturnix japonica)]. In three breeds of domestic quail (Coturnix coturnix japonica) with different zootechnical characteristics, heterozygote frequencies of egg white and serum proteins are variable. Data are briefly explained by the genetic homeostasis theory.


Changes in the water and sodium permeability of amphibian skin in the presence of the efferent plasma of "dehydrated dog kidney". Two kidneys of the same dog were perfused separately with the same blood pool. After the control periods one of the kidneys was replaced by another one taken from a previously dehydrated dog. The presence of the "dehydrated kidney" resulted in a significant and almost immediate decrease in creatinin clearance of the contralateral perfused kidney. The Ccreat increased after the removal of the dehydrated kidney. Plasma samples were taken during the control, transplantation and recovery periods and their activity on the water permeability (osmotic water flow) and sodium transport was examined in amphibian skin. The presence of the plasma taken during the transplantation period resulted in a highly highly significant decrease in the osmotic water transport without any change in the sodium transport. This decrease was reversible, at least partly, after replacing the experimental plasma by a control one. These changes were not observed when a "desalted kidney" was used instead of the "dehydrated" one. These results suggest that in the status of dehydration the kidney releases some humoral factor or factors, which would induce a decrease in the measured GFR, possibly by a direct or indirect effect on membrane, probably glomerular membrane-permeability.


Heterogeneity of bile pigments formed by the breakdown of haemoglobin in the isolated male-dog kidney. Normothermic perfused isolated male-dog kidneys formed radioactive bile pigments by the breakdown of radioactive haemoglobin prepared from [2-14C]glycin. After column chromatographic separation and preparation of dipyrrolic azopigments, 86.3 +/- 2.2% of the bile pigments seemed to be conjugated bilirubin. Thin-layer chromatographic separation of the azopigments of ethyl anthranilate revealed a good correlation between photometric scanning, radiochromatographic scanning and the radioactivity of the azopigments scraped off the thin-layer glass plates and counted in a liquid scintillation counter. Although the same heterogenity of the azopigments was observed as in dog bile, the isolated male-dog kidney formed significantly less alpha2- and significantly more gamma-fractions.


Coagulocyte alterations in clotting hemolymph of Carausius morosus L. 1. The structural changes in the coagulocytes of Carausius morosus during hemolymph coagulation in vitro have been studied under the PCM and in the TEM. 2. In agreement with former PCM  observations on Carausius morosus, the coagulocytes are the only hemocytes to induce coagulation of the plasma. Immediately or after a few seconds upon withdrawal of the hemolymph, their structural changes consist of a considerable enlargement of the perinuclear cysterna and of direct ejection into the plasma of nuclear and cytoplasmic substances through microruptures of the cytoplasmic membrane. The other categories of hemocytes do not contribute to the plasma coagulation. Their structural alterations take place without breakage of the cytoplasmic membrane when the plasma reactions are already established. 4. These plasma reactions appear in the form of circular islands of granular material around the coagulocytes, of extension of the coagulum in the channels between the islands and of transformation of the clot into a network of threads. 5. As reported in other studies in the TEM, no specific organelle characteristic of the coagulocyte ultrastructure could be found in the coagulocytes of Carausius. 6. Owing to the absence of any specific structural criterion of identification, the results suggest that the functional difference between coagulocytes and the other categories of hemocytes as regards coagulation of the plasma might be caused in part by differences of permeability of the cell membranes.


Intermediary hepatic metabolism of rat after oral medium chain triglyceride load. Hepatic ketogenesis was studied in rats given medium chain triglycerides (MCT). Acetyl-CoA accumulated in the liver, indicating a very rapid beta-oxidation of medium chain fatty acids. Citrate level increases. Ketogenesis is strongly enhanced. Cytoplasm and particularly mitochondria are more reduced after MCT. This may explain the very high increase in hepatic malate. Under our experimental conditions, there appears to be a significant linear relationship between the hepatic acetyl-Coa level and those of total ketone bodies and malade respectively.


[Comparative study of different extraction methods and assays of tannins in some pteridophytes]. Various processes of extraction and quantitative analysis of a condensed tannin in a plant extract, which also includes some chlorogenic acids, have been examined. 60% methanol, at 50 degrees C, proved the most efficient extraction solvent. Several methods of analysis have been tried. The measure of the colour intensity obtained by the action of sulphuric vanilline on flavanols cannot be used because it depends on the tannin condensation stage. It is impossible to separate tannin from chlorogenic acids  using the methods of adsorption by skin or nylon powders, or precipitation by polyvinylpyrrolidone. Only paper chromatography, followed by the distinct elution of the various phenolic compounds, allows the tannin evaluation by subtraction; but owing to the variability of the results, many more experiments are necessary. Some other processes are being studied.


Selective radioimmunoassays for human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG). Highly specific radioimmunoassay systems were developed for measurement of hLH and hCG using antisera purified by affinity chromarography or simple adsorption to select the antibodies reacting specifically with either gonadotropin. Such systems permit specific measurement of lLH and hCG in samples containing both. These assays are suitable for various clinical and physiological studies, particularly, study of pituitary functions in the presence of chorionic or trophoblastic secretion.


Supersensitivity to ACh in muscles after prolonged nerve block. Sciatic nerves of rats and tibial nerves of rabbits were kept anaesthetized in situ for periods of 3-11 days by applying silastic cuffs containing lidocaine base or marcaine hydrochloride. To insure a more uniform release of the rapidly diffusing lidocaine base, the drug was contained in compartments at some distance from the nerve and the cuffs were covered with polystyrene. The completeness of anaesthesia and the functional state of the nerve were tested by stimulating the exposed nerves proximal and distal to the cuff and by observing the behaviour of the muscle prior to killing the animals. The ACh sensitivity was tested by electrophoretic application of ACh from micropipettes and by recording the results changes of the resting membrane potential in individual muscle fibres. The ACh sensitivity was found to be present in the extrajunctional area of all muscle fibres including those displaying miniature end-plate potentials. These was no difference between the behaviour of muscles from rats and rabbits and between the action of lidocaine base and marcaine hydrochloride. Previous reports on the absence of extrajunctional ACh sensitivity in muscles of rabbits whose nerves had been treated by lidocaine base were explained by a relatively rapid loss of the drug from the usual type of nerve cuffs (more than 70% of the drug lost in one day), permitting a premature recovery of the nerves from anaesthesia.


[Method to study intestinal tonus in dogs. Preliminary results]. An experimental procedure is described for studying the muscular tonus of a Thiry-Vella isolated jejunal loop, in the conscious dog. Our results demonstrate a reduction of the muscular tonus after the administration of atropine and its increase under the influence of prostigmine. No constant effect was observed after a standard meal.


Coronary venous flow and O2 saturation during transitional phases between various cardiac rates. The instantaneous and continuous interrelationship between coronary blood flow and coronary venous O2 saturation was determined during transient periods following abrupt rate change in the electrically paced canine heart. Through a catheter in the coronary sinus, O2 saturation was continuously monitored using a fibreoptics technique, and venous flow was measured with an electromagnetic flowmeter. Various patterns of change in flow and O2 saturation were observed depending both on the absolute values of the cardiac rates as well as on the relative difference between them during changes from one rate to another. Whereas elevation of coronary flow was monophasic when the magnitude of heart rate change was below 75 beats per minute, a drop in flow was observed preceeding its elevation when the difference was greater. At high rates further increase in rate caused either no alteration or led to a monophasic drop in flow during the transitional period. Changes in O2 saturation were observed only when heart rate difference exceeded 60 beats per minute. Between 60-90 beats per minute O2 saturation remained steady except during the transient rate elevation, ending in a lower steady state O2 saturation. The results indicate that both O2 saturation and coronary flow change with heart rate initially because of mechanical consequence of the increased rate on the myocardium, and later according to its new metabolic needs also manifested by changed O2 extraction.


Changes induced by angiotensins and prostaglandin E2 on the release of transmitter from isolated perfused rabbit kidney during periarterial stimulation. The influence of A II and PGE2 on the rise of perfusion pressure induced by periarterial stimulation and NA were studied in the rabbit isolated perfused kidney. Periarterial stimulation produced an increase in perfusion pressure and the venous outflow superfusing the rabbit aortic strip caused the muscle to contract. Both effects were found to be frequency dependent. NA induced similar effect when given into the renal artery. A II and its N-terminal analogs produced equal potentiation to periarterial stimulation without altering the effect of exogenous NA when added to the perfusion medium. DMGIA II which is a competitive inhibitor of A II inhibited the potentiating affect of A II. PGE2 also inhibited the effect of A II without altering the effect of exogenous NA. Addition of aspirin to the perfusion medium caused a potentiation to periarteral stimulation but did not change the effect of NA. A II added to the perfusion fluid containing aspirin still caused potentiation. From these results it was concluded that: (i) A II-induced potentiation to periarterial stimulation is mediated via specific receptors and probably due to facilitation of the release of transmitter from sympathetic nerve ending. (ii) PGE2 inhibited the release of transmitter. The effect of A II and PGE2 seemed to be mediated by independent mechanisms.


[Effect of injection of synthetic Hyalophora cecropia juvenile hormone in Locusta migratoria L]. The synthetic racemic C18 Hyalophora cecropia juvenile hormone (JH-I) is injected at does of between 10 and 200 mug/animal at the end of the fourth instar of Locusta migratoria. The effects on mortality, length of the fourth and fifth instars, pigmentation and morphogenesis are reported. Higher doses of JH-I produce a higher mortality than lower doses. But mortality can also occur following the injection of oil which sometimes takes place only a few hours before the ecdysis. In no case is JH-I able to shorten the length of the instar. Many animals moult at the same time as the controls, but some of them, both in the fourth and fifth instars, show an important increase in the length of the instar because of an inhibition of the ecdysis. The effect of JH-I on pigmentation is very important and doses higher than 50 mug/animal present a greater effect than an implantation of one pair of corpora allata, both on the number of insects which turn green and on the intensity of this green pigmentation. At the metamorphosis the larvae injected with JH-I produce imperfect imagos and supernumerary larvae, the number of which depends upon the dose. Nevertheless the morphogenetic effect is considerably lower than that of one pair of corpora allata. We have reason to think that this is only due to the time of injection and not to the activity on morphogeneis of the injected hormone. JH-I is injected at the dose of 200 mug in young females which were allatectomized beforehand to prevent oocytes maturation. The hormone completely counter-balances the lack of the corpora allata and some days after the injection the oocytes are in the same state of development as those of the controls. All the results indicate that the synthetic racemic C18 juvenile hormone of Hyalophora cecropia shows a quite similar activity to the secretion of the corpora allata on Locusta migratoria although it has been said for some time that this hormone was not the principal one in locusts.


[The function of different lymphocyte groups and methods for their separation (author's transl)]. A review is given of the participation of T- and B-lymphocytes in the three most important lymphocyte functions: antibody formation, cellular effector reactions, especially cytotoxicity and regulator functions, with particular reference to the concept of "lymphocyte population". The most commonly used methods for the separation of T- and B-lymphocytes and of lymphocytes with the same function are described.


Alpha-antitrypsin deposition in primary hepatic carcinomas. With the use of immunoperoxidase methods, paraffin-embedded hepatic tissue from 21 patients with primary hepatic carcinoma was tested for alpha-antitrypsin (AAT) deposition and was compared for sensitivity with the PAS reaction. Specific AAT immunoreactivity was present in tumor cells in half of the test cases, either alone or in combination with positive nonneoplastic hepatocytes. While the PAS findings generally parralleled the specific immunohistochemical studies, the latter technique exhibited greater sensitivity in AAT detection. Cytoplasmic tumor cell inclusions of three types were identified, but only one type, the solid, was found to represent immunoreactive AAT deposits. These findings confirm as association between hepatic deposition of AAT and the occurrence of primary liver carcinoma. Further prospective serum and tissue studies on patients with hepatoma are needed to extend these observations.


Isoantigen loss in cervical neoplasia. Demonstration by immunofluorescence and immunoperoxidase techniques. Tissue sections from normal uterine cervix and from uterine cervices with dysplasia, carcinoma in situ, and invasive cancer were studied in 70 instances using immunofluorescence and immunoperoxidase techniques for demonstrating the presence of isoantigens A and B. Isoantigens were demonstrated in normal squamous epithelium. Antigens were lost in foci of invasive cancer, were considerably reduced or lost in areas of classic carcinoma in situ in all cases, and were reduced in most cases of dysplasia. These observations confirm those obtained with the specific red cell adherence test. The results suggest that immunofluorescence and immunoperoxidase techniques are sensitive, reproducible, easily performed methods for demonstrating the loss of isoantigens in cervical neoplasia.


[Reaction of the liver to the effects of hexachlorocyclohexane (experimental study)]. The effects of hexachlorocyclohexane (HCHCH) on the weight of the liver, content of protein, glycogen, total lipids as well as morphological and ultrastructural changes in the liver were studied. It was established that HCHCH caused a considerable increase in the weight of the liver, as well as in the levels of protein, glycogen and total lipids, which was particularly pronounced following prolonged administration of HCHCH in the dose 1/50 LD50. Large doses of HCHCH (1/3 LD50) brought about marked dystrophic changes in the liver similar to the type of hydropic degeneration, accumulation of fatty drops in hepatocytes Application of a dose 1/50 LD50 of HCHCH resulted in an increased size of hepatocytes, dystrophic changes being noted only in individual cells. Histochemisally there were revealed high levels of RNA and glycogen in hepatocytes. An-Electron-microscopy study of hepatic cells showed the most manifested changes in the endoplasmatic network of hepatocytes, which were seen in an enlargement of the elements of the smooth endoplasmatic reticulum. A suggestion was put forward that the changes indicated above were associated with a stimulating effect of HCHCH on the microsomic system of hepatocytes, i.e. on the enzymatic systems localized in the smooth endoplasmatic network.


[Development of the ideas of N. N. Anichkov at the present stage of our knowledge about atherosclerosis]. The article discusses the main elements of the infiltrative-combination theory of atherosclerosis, which was formulated and developed by N. N. Anichkov. The further development of the main principles of this theory at the current stage of medical science is elucidated. Analysis of the mechanisms of penetration of lipids into the vascular wall, the role of metabolism of the main substance and plasmic proteins at the initial stages of atherosclerosis shows that the new data obtained are not at variance with the fundamental principles of the N. N. Anichkov's theory but on the contrary, supplement it. Evidences on the role of immune complex of beta-lipoproteid-gamma-globulin in the development of atherosclerosis are also in complete accord with this theory.


[Vascular hyalinosis in diabetic microangiopathy]. A histochemical and immunoluminescent investigation of hyalin of arterioles ans small arteries of the kidney, spleen, pancreas and skin were carried out. In 20 patients with diabetes mellitus of a various degree of gravity and duration hyalinosis represented a manifestation of microangiopathy; it was particularly pronounced in the vessels of the spleen and kidney, and in a lesser degree--in the vessels of the pancreas and skin. The intensity of hyalinosis was directly proportional to the duration of the disease; Immunomorphologically, hyaline masses devealed globulins, gamma M-, gamma G-immunoglobulins, complement, albumins, beta-lipoproteins, fibrinogens, which indicates to a hematogenic nature of the vascular hyalin. The presence in hyalin of G an M-immunoglobulins and complement may testify to the participation in its formation of immune mechanisms, together with metabolic ones.


Experimental specific memory reaction to cornea, lens, and retina antigens. Lymphocytes from rabbits immunized against various ocular extracts were challenged in vitro by the immunizing extract, by other ocular extracts from the same species, and by analogous extracts from other species. Immunological memory reaction, demonstrated by the blast transformation phenomenon, was found toward organ-specific and species-specific antigens. The organ cross-reactivity was greatest when cornea and lens were tested. Immunization of rabbits against calf cornea with extract emulsified in Freund adjuvant injected into the hind foot-pads induced the appearance of specific memory cells in popliteal lymph nodes, spleen, and blood. Immunization by interlamellar xenografting has shown that memory lymphocytes are mainly localized in the preauricular lymph nodes and are nearly absent from spleen and popliteal lymph nodes. These findings indicate the important role of the local immunological mechanism of the eye.


Cardiac monitoring during exercise tests in the horse. 3. Changes in the electrocardiogram during and after exercise. Changes that occur in the equine ECG during and after exercise have been described and compared with resting ECG's obtained from the same horses. When the speed of work equals or is greater than "three-quarter pace" (i.e. "even time" or 200 metres in 15 seconds) the high heart rates developed cause the loss of ECG waveforms that are readily discernible at rest. Although differences in the waveforms between horses with normal and abnormal resting ECG's have been observed, the more definitive useful information is the fact that abnormal horses showed significantly higher heart rates than normal horses at a particular speed of work. When this occurs in combination with a resting ECG classified as borderline or abnormal and the exercise ECG shows positive, low amplitude, broad rounded T waves or transient bursts of sine-waves suggestive of ventricular flutter, racing performance is likely to be impaired. Although more work needs to be done and improvements in technology are necessary, the exercise ECG has potential for improving understanding of physical fitness and the significance of resting cardiac abnormalities.


Investigations on d-galactosamine hepatitis after pretreatment with alpha-hexachlorocyclohexane. D-Galactosamine hepatitis cannot be induced in rapidly replicating liver tissue at various times after induction of proliferation. Proliferation was induced by administration of alpha-hexachlorocyclohexane. The morphological features of galactosamine hepatitis do not appear or are very mild. The onset of DNA synthesis is delayed to about 12 hrs as also shown in partially hepatectomized rats.


[Experiences at the Blood Transfusion Center of the Socialist Republic of Serbia in providing aimed transfusions]. The development of modern medicine causes the more and more needs of blood in providing the complete health care of the patients. The increased need can be satisfied only by the rational use of blood on the principle of selective transfusions. In this work our present possibilities are given, in the separations of certain blood components, their good and bad effects, as well as the indications for use in some pathological states.


[Subtypes of serum hepatitis B antigen]. Rheophoresis improved gel diffusion technique was used for the determination of subtypes of HB antigen (HBAg) according to D (ad + y) and Y (ay +, d--) specificity. Although the relative small number of carriers of antigen was investigated, it seems that the distribution of subtypes is somewhat different from the results found by the american and scandinavian autors and is more similar to the results of Hadziyannis (Greece) and Mosley in Israel. The specifity of serum containing HB antibodies was also investigated. Five sera containing antibodies, revealed both ad and ay subtypes by the counter electrophoresis method, but two of them showed a significantly stronger precipitation line with subtype ay.


Electrocardiographic antecedents of primary ventricular fibrillation. Value of the R-on-T phenomenon in myocardial infarction. Primary ventricular fibrillation was seen in 20 of 450 consecutive patients (4-4%) admitted within 24 hours after the onset of acute myocardial infarction. Compared with patients without primary ventricular fibrillation they showed a lower mean age group and a higher incidence of anterior infarction. Warning ventricular arrhythmias preceded primary ventricular fibrillation in 58% of cases. However, warning arrhythmias were also present in 55% of patients without primary ventricular fibrillation. The following mechanisms of initiation of primary ventricular fibrillation were seen. 1) In one patient, it was initiated by supraventricular premature beats showing aberrant intraventricular conduction. 2) In 2 patients, ventricular tachycardia degenerated into primary ventricular fibrillation. 3) In 17 patients, it was initiated by a ventricular premature beat; in 10 of these, the premature beat showed early coupling (RR/QT less than 1--the R-on-T phenomenon). However, ventricular premature beats showing the R-on-T phenomenon were also observed in 49% of patients without primary ventricular fibrillation. In 7, primary ventricular fibrillation was initiated by a late-coupled ventricular premature beat (RR/QT greater than 1); in 2, the very late coupling resulted in a ventricular fusion beat. The study suggests that warning arrhythmias and the R-on-T phenomenon are poor predictors of primary ventricular fibrillation in acute myocardial infarction. The observation that 41% of primary ventricular fibrillation was initiated by a late-coupled ventricular premature beat suggests that ventricular vulnerability during acute myocardial infarction may extend throughout most of the cardiac cycle and is not necessarily confined to the QT interval.


Association of ventricular extrasystoles and ventricular tachycardia with idioventricular rhythm. Patients with acute myocardial infarction were monitored for ventricular arrhythmias in the first 48 hours. Idioventricular rhythm (rate less than 100/minute) occurred in 35 out of 224 patients (15.6%) during the first day and in 13 out of 192 patients not receiving treatment on the second day (6.8%). This arrhythmia was frequently preceded by late ventricular extrasystoles, which often showed variation of their coupling intervals to the preceding QRS. Double ventricular extrasystoles separated by larger than or equal to 600 ms were also precursors of idioventricular rhythm. Idioventricular rhythm at times could be described as an escape rhythm, but on other occasions it was undoubtedly an accelerated rhythm. Spontaneous changes in the idioventricular cycle length were frequent on single one-minute electrocardiographic recordings. The rate of the dominant rhythm in patients with episodes of idioventricular rhythm was significantly slower than the heart rate of patients without this arrhythmia. Idioventricular rhythm was more frequent in patients with inferior infarction. Idioventricular rhythm sometimes preceded ventricular tachycardia but there was only a significant association between ventricular tachycardia and idioventricular rhythms with rates of over 75/minute. Irregular idioventricular rhythm frequently accelerated to ventricular tachycardia. It is suggested that the term benign idioventricular rhythm be reserved for those rhythms below 75/minute, and that the term rapid idioventricular rhythm should be used for rhythms between 75 and 120/minute. The rate of idioventricular rhythm is related to the probability of development of life-threatening ventricular arrhythmias during the first 48 hours after myocardial infarction.


Second heart sound after pulmonary arterial banding operation. After a pulmonary arterial banding procedure the phonocardiograms of 38 patients were correlated with haemodynamic and angiographic findings. Twenty-four patients had uncomplicated ventricular septal defect, 2 had single ventricle, 5 had transposition of the great arteries, 5 had atrioventricular canal defects, and 2 had coarctation of the aorta and ventricular septal defect. P2 was separated from A2 by less than 40 ms in 10 of the 11 patients with high pulmonary vascular resistance. Of 27 patients with nearly normal pulmonary vascular resistances and distal pulmonary artery pressures less than 50/20 mmHg (6-7/2-7 kPa), 24 had A2-P2 intervals of over 40 ms. A narrow A2-P2 interval with a satisfactory band was found in 2 patients with large right-to-left shunts. A2-P2 interval did not change appreciably with age, and this measurement is a useful guide as to the effectiveness of pulmonary artery banding by one year after operation. If this interval is less than 40 ms, repeat catheterization should be carried out as such patients may have persisting pulmonary hypertension and an increased pulmonary vascular resistance.


Human preleukaemia cell culture studies in sideroblastic anaemia. Cell structure abnormalties are found in acute leukaemia and preleukaemic states. Studies on bone marrow cells and peripheral leucocytes of 4 patients with idiopathic acquired sideroblastic anaemia showed patterns in cell culture similar to those reported in acute leukaemia: 2 of these patients later developed leukaemia. Other patients with idiopathic, secondary or congenital sideroblastosis showed no such cell culture abnormalities, and none developed leukaemia. Studies such as this suggest that cell culture methods detect altered cellular function preceding overt leukaemia and that these abnormal findings may be helpful in the evaluation of patient groups with an increased incidence of leukaemia.


The sensitivity to cytotoxic agents of the EMT6 tumor in vivo. Comparative response of lung nodules in rapid expotential growth and of the solid flank tumour. Experiments are described in which dose-response data have been obtained for EMT6 mouse tumour cells growing in vivo and exposed to various cytotoxic agents. A comparison has been made of the response of solid tumours in the flank and of rapidly growing lung nodules. The results are discussed with regard to their cell kinetic implications and compared with our results for the chemosensitivity of EMT6 cells in exponential and plateau phase growth in vitro.


A quantitative study of the localization of atherosclerotic lesions in the rabbit aorta. A quantitative study of the size and location of early sudanophilic lesions has been carried out on the aortae of five rabbits. The atherosclerotic lesions were induced by feeding an average of 114 (+/- 3.7) egg yolks over an average period of 83 (+/- 1.7) days. The aortic lesions were visualuized by gross staining with Sudan III and measured by the polar coordinate method. The lesions were almost entirely around orifices: their size was directly proportional to the area of the associated ostium (P less than 0.005). The sudanophilic deposits were located downstream from the ostia in areas believed to experience high shear stresses. The area of the intercostal ostia increased as one proceeded down the thoracic aorta (P less than 0.005). A deviation from the distal distribution pattern was observed where local flow and shear stresses were disturbed. The coronary lesions completely surrounded the ostia, the coeliac lesions had significant proximal components and the left renal and inferior mesenteric lesions were skewed to the right. The study suggests that hemodynamic forces and particularly high shear stress is of considerable importance in both the initiation and localization of early atherosclerotic lesions.


Tumour-associated immune responses and isolated carcinoembryonic antigen and alpha feto-protein levels related to survival in ovarian cancer patients. The presence of a tumour-associated immune response in 37 patients with ovarian cancer as assessed by blastogenesis (lymphocyte transformation) evoked by ovarian cancer cell extracts, has been correlated with survival following the test. The difference in these responses is unlikely to be accounted for on the basis of general impairment of cell-mediated immuno-competence. Serum carcinoembryonic antigen (CEA) was also determined in 27 ovarian cancer patients to assess its prognostic significance. Raised CEA levels and absence of blastogenic response to tumour cell extract during relapse are associated with a worse prognosis but neither of these parameters are significant in remission. Possible applications of these findings to the clinical management of ovarian cancer patients are discussed. Serum alpha feto-protein levels measured by radioimmunoassay were not found to be raised in any of the 32 ovarian cancer patients in whom it was measured.


DNA polymerase activity in plasma from leukaemic guinea-pigs. Measurement of DNA polymerase in leukaemic guinea-pig plasms reveals the presence of low levels of sedimentable and non-sedimentable enzymic activities. Since the sedimentable DNA polymerase is ribonuclease sensitive, uses poly(C).oligo(dG) as template, and bands in a sucrose density gradient at 1-17 g/ml it is thought to be the GPLV-associated reverse transcriptase. The soluble DNA polymerase is stimulated by ribonuclease and is probably of cellular origin.


Quantitative recovery of lipids from aortas stained with sudan IV. The separation of Sudan IV from lipid extracts of stained aortas can be accomplished by thin layer or column chromatography, but the most convenient method involves adsorption of the dye to activated carbon. The aortic lipids are thus recovered quantitatively.


The effect of bleomycin and pentamycin in combination on the survival of EMT6 mouse tumour cells in vitro and in vivo. The combined effect of bleomycin and the polyene antibiotic, pentamycin, upon the survival of EMT6 tumour cells has been studied both in vitro and in vivo. During growth of the cells as a monolayer in vitro, a very marked potentiation of bleomycin action is seen during the exponential and early plateau phases of growth, but little potentiation occurs in late plateau phase. The effect of exposing the cells to the two drugs consecutively in either order is greater than if the two agents are used at the same time. In vivo, it does not appear that pentamycin can greatly increase the cytocidal effect of bleomycin.


Intimal thickening in arteries of rats treated with synthetic sex hormones. The effect of ethynyl oestradiol and chlormadinone acetate, separately and combined, on the aorta, carotid, mesenteric and renal arteries of female rats was studied using light and scanning electron microscopy. The hormones were injected daily for either 30 or 90 days. The most constant finding was intimal thickening which consisted mainly of areas of subendothelially located smooth muscle cells. Each artery was given a score from 0 to 3 according to the degree of development of these areas. Ethynyl oestradiol, unlike chlormadinone acetate, was associated with sustained significant enhancement of both incidence and degree of intimal thickening. With the combined treatment, the degree of intimal thickening in the 30-day group was not appreciably different from that obtained with oestrogen alone. However, in the 90-day group it was significantly less in arteries other than the aorta, suggesting that the progestogen inhibited the enhancing effect of the oestrogen on the proliferative process in these arteries.


Three approaches to the radioimmunoassay of human beta-thromboglobulin. Three radioimmunoassays for the measurement of beta-thromboglobulin are described. The standard method, using antiserum in solution, could be used to measure plasma concentrations of beta-thromboglobulin with the results available after 2-3 d. The use of a greater dilution of antiserum and tracer, with delayed addition of tracer, resulted in a more sensitive assay suitable for measuring b-thromboglobulin in urine. The use of a solid-couples antiserum under non-equilibrium conditions allowed the measurement of plasma levels of beta-thromboglobulin after an assay incubation time of 1 h. These three radioimmunoassay systems for beta-thromboglobulin cover the likely clinical requirements for the measurement of this platelet specific protein.


Studies on liberation of beta-thromboglobulin from human platelets in vitro. A platelet specific protein, beta-thromboglobulin, is liberated during the preparation of platelet poor plasma. Using combinations of different anticoagulant and anti-platelet compounds, this release can be significantly reduced. The best results were obtained when native blood was collected as soon as possible into a mixture of EDTA, prostaglandin E1 and theophylline and maintained and processed at a temperature between 0 and 4 degrees C. These technical innovations have permitted the use of a radioimmunoassay for beta-thromboglobulin on plasma samples in clinical practice.


Teaching interviewing skills to medical students: a comparison of two methods. A group training course utilizing videotaped material and discussion proved to be as effective as traditional methods in teaching medical students to obtain information from co-operative psychiatric patients. There were also some clear indications that group video training may prove valuable in teaching specific interview techniques, although its full potential in this area has yet to be explored.


Audiotape slide programmes: equipment and production. The number of medical audiotape-slide programmes available has been rapidly increasing. A typical programme comprises a cassette tape and a set of slides. A questionnaire or workbook may also be provided, e.g. in the programmes which may be borrowed or purchased from the Medical Recording Service Foundation. For playback a cassette recorder and projector will suffice or, for economy, a stacking viewer may be used. Automated apparatus is not essential and if used should be of the correct 'USPEC 2' standard. To produce audiotape-slide programmes is more exacting than may be anticipated, and details of the procedure are provided. Programmes should be assessed and upgraded after their production.


Alpha-fetoprotein levels in maternal plasma and amniotic fluid during prostaglandin-induced mid-trimester abortions: the relation to fetal distress and death. Alpha-fetoprotein (AFP) levels were measured in both plasma and amniotic fluid during 43 abortions induced with prostaglandin F2alpha (PGF2alpha) and their relationship to the induction-abortion interval and signs of fetal distress or intrauterine death examined. The mean plasma AFP levels showed a progressive rise after PGF2alpha administration but abnormally high levels were found infrequently until the time of delivery. Feto-maternal bleeding could explain the abnormally high plasma AFP levels which occurred early in a few cases. A shorter induction-abortion interval was found in patients with normal plasma AFP levels. Neither fetal distress nor intrauterine death were associated with abnormally high plasma AFP levels. No change in AFP levels in amniotic fluid was found.


The placental and fetal response to the intra-amniotic injection of prostaglandin F2alpha in midtrimester abortions. The placental and fetal response to the intra-amniotic injection of prostaglandin F2alpha (PGF2alpha) for midtrimester abortions was evaluated. Maternal blood samples for radioimmunological determinations of serum progesterone, human placental lactogen (HPL) and alpha-fetoprotein (AFP) were taken before and serially until 24 hours after the injection of 40 mg (nine patients) or 75 mg (seven patients) of PGF2alpha. Fetal heart action was monitored ultrasonically during the same period. The average induction-abortion interval was 22-6 hours and was indepedent of the dose of PGF2alpha used. The mean progesterone and HPL levels showed a significant fall half an hour after PGF2alpha. The constant decreases began only at 5 hours and later. The mean AFP levels increased rapidly after 5 hours following injection of PGF2alpha. The feral heart stopped at a mean of 10-4 hours (range 0-5 to 25 hours) after the injection of PGF2alpha. No relationship between the time of fetal death or rises in AFP and the induction-abortion interval could be detected. The primary hormonal response of the placenta and the efficacy of exogenous PGF2alpha were not related, indicating the possibility that the abortifacient property of PGF2alpha is not related to its primary effect on placental hormone synthesis or release.


A serial study of pregnancy proteins in primigravidae. The plasma concentrations from four 'pregnancy proteins' and three steroid hormones have been measured throughout pregnancy in 15 primigravidae. Two of the proteins, human placental lactogen (HPL) and pregnancy-specific beta1-glycoprotein (PSbetaG), are specific for pregnancy and correlate well with the stage of gestation. It is suggested that measurement of PSbetaG may be useful in assessing feto-placental wellbeing. Neither of the pregnancy-associated proteins, steriod-binding beta-globulin (SBbetaG) and pregnancy-associated alpha2-glycoprotein (alpha2-PAG), correlated with the length of gestation although, near term, plasma alpha2-PAG levels appeared to be related to fetal weight. In addition, no correlation has been demonstrated between placental or fetal weight and any of the pregnancy-specific proteins or steroid hormones studied.


Voltage-induced thickness changes of lipid bilayer membranes and the effect of an electrin field on gramicidin A channel formation. The thickness changes of black lipid membranes of different composition after a voltage jump were investigated. In a second series of electrical relaxation experiments the kinetics of channel formation by gramicidin A were measured. The time course of the membrane current was compared with the time course of the thickness change of the membranes. We found that the time course of the current as a consequence of channel formation by gramicidin A did not correlate with the thickness change of the lipid membranes. A possible direct influence of the electric field is discussed.


The solution behavior of the bovine myelin basic protein in the presence of anionic ligands. Binding behavior with the red component of trypan blue and sodium dodecyl sulfate. The interaction of the azo dye (2,3'-dimethyldiphenyl-7-azo-8-amino-1-napthol 3,6-disulfonic acid (TBR) and sodium dodecyl sulfate with the bovine myelin basic protein has been studied using absorbance, circular dichroism and 220 MHz PMR spectroscopy. Additional analyses of the binding reaction were carried out using light scattering, ultracentrifugal and electrophoretic techniques. A procedure for preparing pure TBR was developed. A modified structure for this synthesized TBR has been suggested. The mechanism of TBR binding to the myelin basic protein was found to be metachromatic. In addition, the interaction of TBR with the basic protein which gives rise to aggregation of the dye bound species was found to be analogous to the model proposed by Schwarz, G. and Seelig-Löffler, A. ((1975) Biochim. Biophys. Acta 379, 125-138) to explain the binding of acridine orange with poly (alpha-L-glutamic acid). PMR spectral analyses suggested that arginine residues provide the majority of primary sites of attachment on the basic protein for TBR. The effect of sodium dodecyl sulfate binding with the bovine myelin basic protein was found to induce a minimal change in the conformation of the protein. The induction of only about 20% alpha helial structure could be demonstrated and the binding was reversed by raising the solution temperature to 73 degrees C. The difference in the observed behavior of basic protein arising from TBR binding as opposed to the binding of sodium dodecyl sulfate is viewed as resulting from two different binding mechanisms. The binding behavior of TBR is primarily a consequence of charge-charge interaction while the binding effects of sodium dodecyl sulfate are a consequence of hydrophobic interaction. The sodium dodecyl sulfate binding acts as a shield which limits charge-charge interaction in the basic protein molecule thus preventing aggregate formation while TBR imposes no such restraints.


Binding of 17beta-estradiol by variants of alpha-fetoprotein in rat amniotic fluid. Two variants of alpha-fetoprotein in rat amniotic fluid were separated by their different affinity for concanavalin A-Sepharose, which selectively binds alpha-D-manno-pyranosides and alpha-D-glucopyranosides. Both forms had the same mobility upon polyacrylamide gel electrophoresis. The binding of 17beta-estradiol per mg of alpha-fetoprotein, determined both immunologically and electrophoretically, was the same for both variants. These results indicate that a specific carbohydrate portion of the molecule is not necessary for steroid binding.


The high molecular weight proteins released from cultured cells. Studies have been performed with the serum-free culture medium taken from several fibroblast monolayer culture lines. A high molecular weight protein fraction was separated from the concentrated medium by sucrose density gradient centrifugation. Polyacrylamide gel electrophoresis was used to assess the degree of purification obtained. In the electron microscope the negatively stained high molecular weight proteins were found to closely resemble the alpha2-macroglobulins. The suggestion that these proteins from cultured cells resemble the cylindrical protein complex isolated from mammalian erythrocyte ghosts is not supported by this study. The results are discussed in the light of the extensive literature now available on the electron microscopy of high molecular weight proteins.


Enzymic and immunochemical properties of lysozyme. XVI. A novel synthetic approach to an antigenic reactive site by direct linkage of the relevant conformationally adjacent residues constituting the site. Previous studies from this laboratory on the immunochemistry of specific chemical derivatives of native lysozyme and of the two disulfide peptide 62-68 (Cys 64-Cys 80) 74-97 (Cys 76-Cys 94) (i.e. (SS)2-peptide), have established an antigenic reactive site to comprise the spatially contiguous surface residues: Trp 72, Lys 97, Lys 96, Asn 93, Thr 89 and Asp 87. In the present work, the identity of the site was verified by an entirely different and novel approach. The aforementioned amino acids were linked directly into a single linear peptide with an intervening spacer where appropriate and substituting phenylalanine for tryptophan (i.e. Phe-Gly-Lys-Asn-Thr-Asp). This peptide (which does not exist in native lysozyme but simulates a surface region of the protein) possessed a remarkable inhibitory activity towards the reaction of lysozyme with its antisera. The immunochemical reactivity of the peptide was equal to the maximum expected reactivity of the site (i.e. a third of the total antigenic reactivity of lysozyme). These findings define quite conclusively and accurately the reactive site which is clearly composed of spatially adjacent residues that are distant in sequence reacting as if in direct linear linkage. The unequivocal establishment of this concept indicates that antigenic sites need not always be composed of residues in direct peptide linkage in the sequence. The nature of the site may depend on the protein. This unorthodox attack at the problem provides a novel and powerful approach for final delineation of the antigenic reactive sites (and perhaps other types of binding sites) in native proteins, following the completion of accurate narrowing down by chemical methods.


Gramicidin S-synthetase. Preparation of the multienzymic complex with a high specific activity. A new purification procedure for the multienzyme of gramicidin S-synthetase has been developed. In vitro proteolysis with partial inactivation is suppressed by protease inhibitors EDTA, phenylmethylsulfonylfluoride, and fast preparation methods during initial separation steps. Activity has only been assayed by the total reaction of gramicidin S-synthetase, not by partial reactions of amino acid activation. The assay has been improved by evaluation of inhibitory concentrations of buffers, salts, and the product gramicidin S. It has been demonstrated that the rate of peptide synthesis in extracts containing both enzymes of gramicidin S-synthetase depends on protein concentration in a second order function. The multienzyme or heavy enzyme has been purified about 1400-fold to a specific activity of 24 nM/min per mg of protein, and the relation of this activity to the calculated in vivo activity is discussed.