[Use of immunosorbents for determining antibodies to histaglobulin]. A determination was made of antibodies to histaglobulin and gamma-globulin with the aid of immunosrobents in 178 sera of children suffering from allergic diseases, during the histaglobulin therapy. Results of investigations showed antibodies to histaglobulins to be absent in children untreated by this preparation. But they appeared in 57% of cases after the first course of histaglobulin treatment, and their incidence and their average level increased with an increase in the number of the courses of treatment carried out. gamma-Globulin antibodies were found at the initial condition in 42% of cases; this percentage rose to 68 after the first course of histoglobulin treatment. The authors believe that determination of histaglobulin antibodies during the treatment with this preparation could serve as an auxiliary immunological criterion of the efficacy of histaglobulin therapy.


[Spread of viral hepatitis in organized children's collectives and the methods for its early diagnosis]. The authors present the results of observations over 407 children aged from 2 months to 16 years from the foci of viral hepatitis in children's collective bodies. During the quarantine a determination was made in children of the glutamic-pyroracemic, glutamic-oxalic transaminases (GPT and GOT, respectively) and of the hepatitis B antigen (HBAg). A necessity of using the enzymatic tests for the purpose of early diagnosis of viral hepatitis was shown, since 84% of the cases developing in the next focus coursed as an unicteric form without any markked clinical signs; HBAg was revealed in 6.1% of the children examined. A complex examination of the personnel and of the persons who came in contact with the patients with viral hepatitis showed the ways of spread of hepatitis B in a collective body; it was found that the viral hepatitis B infection took place both by parenteral and enteral routes. The expediency of active observation over the children, recipients of blood and plasma, with determination in them of the activity of the enzymes and HBAg for early diagnosis of parenteral infection was substantiated. It was also shown that the incidence of the unicteric forms of viral hepatitis in a focus of infection depended not on the periods of gamma-globulin administration but on the age of children who contracted the infection. Thus, the prevalence of the unicteric forms of the disease over the icteric ones in children under 3 years of age was more pronounced than in older children.


[Cultivation of Neisseria meningitidis serogroup A on semisynthetic liquid nutrient media]. In cultivation of meningococcus of serological group A in fluid semisynthetic medium of simple composition prepared on the basis of purified acid casein hydrolysate with profound splitting there were obtained microbial cultures with a density of 4-5 x 10(9) microbial cells per 1 ml after 20-24 hours of cultivation with shaking. Alkalinity of the medium increased (to pH 8.0-8.2 during the stationary phase) with increase of the microbial cell concentration. A study of the accumulation of group-specific thermostable polysaccharide antigen in dynamics of meningococcus cultivation on semisynthetic medium tested showed the preparations obtained by alcoholic precipitation to be colourless and to contain much antigen (by inhibition of indirect hemagglutination), particularly at the phasees of negative growth acceleration and at the stationary phase. The suggested fluid semisynthetic medium of simple composition could be used for production of diagnostic and prophylactic meningococcus preparations belonging to the serological group A.


[The reactivity of lupus erythematosus serum with native and denatured DNA of various origins]. The serum of a female suffering from Lupus erythematosus visceralis was investigated by complement fixation for the reaction with native and denatured DNA's of various base composition. The reaction with native DNA is independent on the (G+C)-content of the DNA. It is apparent that the responsible antibodies react with determinants of the helical conformation, which are identical in the various DNA-molecules. Quantitative differences are found with denatured DNA's. The strongest complement fixation is observed with (G+C)-rich denatured DNA. The reaction with denatured DNA is only partially inhibited by DNA digest. These antibodies obviously react with sequential determinants containing bases. Therefore, they are induced by a different mechanism of sensitization.


[Methods for the separation of lymphocytes]. The paper reviews the currently available methods for lymphocyte separation, with particular reference to their effectiveness. Procedures based on density and size, such as density gradient centrifugation and sedimentation and size filtration on columns, allow accumulation of lymphocytes of different degree of differentiation, but do not permit any quantitative separation of distinct lymphocyte populations, because density and size of cells are properties  strongly varying with the degree of development and physiological state of the cells. Differences of the cells' net potential cause differential adhesion of lymphoid cells to glass or other materials, and lead to varying migration speeds in the electric field. Adherence columns afford only partial separation of T and B cells, whereas favourable results have been obtained by preparative cell electrophoresis. Special membrane structures, such as differentiation antigens including membrane-bound immunoglobulins, cell receptors and transplantation antigens make possible a specific separation of lymphocytes. Essentially, the following 5 methods are being used: 1. Cytolytic treatment of the cells with antisera against differentiation antigens in the presence of complement. 2. Rosette separation 2.1 Rosette formation with sheep erythrocytes (SE) or with antigen-coaded SE for the isolation of antigen-binding lymphocytes. 2.2 Rosette formation by antigen-antibody-complement complexes (B rosettes) 2.3 Rosette formation with SE by human T lymphocytes (T rosettes) 2.1--2.3. Separation of the rosettes from the free lymphocytes by centrifugation or sedimentation.


[Original techniques for biliary surgery (author's transl)]. 1. Aspiration of intrahepatic stones. This uses a catheter designed for selective aortography with a special probe, short and provided with a soft cap. The instrument is inserted in the hepatic canal to the contact of the stone, under TV guidance. Strong suction allows extraction. 2. Hepato-gastric prosthesis as palliative. Treatment for cancer of the biliary tract. A. Pezzer sound is placed in a dilated hepatic canal at one end, the other being fixed in the stomach. This is an ultimate solution for far advanced cases, that can only suppress the itching.


Treatment of duodenal ulcer with a long-acting synthetic secretin: a pilot trial. In three male duodenal ulcer patients, the effects of a 4-week treatment with a long-acting synthetic secretin -- daily s.c. administrations of 10 clinical units secretin/kg b.w. -- were assessed symptomatically, endoscopically, immunologically, and by means of gastric and pancreatic secretory analyses. Improvement in epigastric pain was noted within a few days following the onset of secretin therapy. In all cases, duodenoscopy revealed complete healing of ulcers after a 2- to 3-week treatment period. Before therapy, cutaneous anaphylactoid reactions due to intradermal injections of secretin could be elicited in each subject; skin reactions were found mitigated after 4 weeks of therapeutic secretin administration thus suggesting some desensitisation. Neither gastric acid secretion nor pancreatic bicarbonate production were substantially influenced by the 4-week course of secretin therapy; consequently, prevention of ulcer relapse is rather unlikely to be achievable with the conditions employed in this study. However, it seems therapeutically promising that the duodenal ulcers under study showed complete healing already after 2-3 weeks of secretin administration.


A controlled study of allergen production in cultures of Dermatophadoides pteronyssinus. Allergenic substances have been isolated from cultures of Dermatophagoides pteronyssinus on wheat germ flakes and powdered yeast; as controls, non-sterile nutrient medium containing no mites, as well as sterile medium, were maintained and extracted under identical conditions. Chemical purification and analyses indicated the occurrence of skin-active allergens in all three preparations. This was confirmed by the assay of skin-reactivity in vivo and complement-inactivating capacity in vitro. Quantitatively, the medium containing bacteria and fungi contained more allergen than the sterile control, but less than the mite-infected medium. The evidence indicates that degradation reactions of nutrient components proceed faster in the non-sterile media than in the sterile control. The results of immunochemical analyses demonstrate that degradation reactions, give rise to the introduction of lysine-sugar structures inot high-molecular weight components from the medium. It is concluded that mites and micro-organisms have a role in allergen formation by promoting degradation reactions among nutrient constituents.


Similar geographical distribution of multiple sclerosis and cancer of the colon. A search was made of all diseases listed in the Annual Statistics of the World Health Organization to determine if any disease had the peculiar geographical distribution of multiple sclerosis. Of the approximately 83 diseases listed, only cancer of the colon was similar in geographical distribution and in the slight preponderance of females dying with the disease. The epidemiology of both diseases is reviewed, particularly with regard to their appearance in certain migrant populations.


Effect of encephalitogenic protein on the migration in agarose of leucocytes from patients with multiple sclerosis. Correlations with clinical parameters, localization of antigenic determinants, and influence of corticotrophin treatment. We have extended the results from our previous investigation of the reactivity of leucocytes from patients with multiple sclerosis and some other neurological diseases to bovine encephalitogenic protein, using the migration in agarose technique. 1. Reactivity (i.e. behaviour in the test deviating from that expected with cells from healthy subjects) is manifested either as an enhancement or an inhibition of leucocyte migration. 2. The incidence of subjects with cells showing reactivity to encephalitogenic protein is higher in the group of patients with multiple sclerosis than in the group of patients with other neurological diseases. 3. Reactivity to encephalitogenic protein is correlated to disease course score; patients with a chronic progression of the disease show the strongest reactivity in the test. 4. The tryptophan-containing region of the encephalitogenic protein appears to be a major determinant of significance for this test. 5. Patients tested during corticotrophin treatment tend to show a higher degree of reactivity in the test than expected-the increased reactivity seems to be primarily directed to the tryptophan-containing region.


[Use of hydroxyethyl starch as a cryoprotective medium during platelet storage at low temperatures]. The effects of HES and HES + DMSO used as cryoprotective media for storage of human platelets in liquid nitrogen and vapor phase of liquid nitrogen were studied. Solution of 6% and 15% HES with molecular weight ranging from 65,000 to 250,000, and 10% DMSO were used. The criteria accepted for evaluation of the efficiency of these cryoprotective media were: 1. platelet counts, 2. participation of platelets in the processes of hemostasis measured in vitro by the ability of platelets to release adenine nucleotides (ATP + ADP) after thrombin stimulation. It was found that 15% HES is a more effective cryoprotective medium than 6% HES. The use of 15% HES + 10% DMSO gave similar results as the use of 10% DMSO alone.


Long-term treatment of ventricular arrhythmias with oral mexiletine. Twenty-four patients with ventricular arrhythmias were treated with oral mexiletine for periods of from one to 16 months (total 10.4 patient-years). In 19 patients arrhythmias were satisfactorily controlled and plasma levels previously shown to be within the therapeutic range were maintained on an 8 hourly regimen. The drug was well tolerated in most patients and regular hematological and biochemical screening revealed no problems of toxicity.


A comparison of the unlabeled enzyme method with immunofluorescence for the evaluation of human immunologic renal disease. The application of the unlabeled enzyme method to immunostaining of human renal biopsy material is described. The specificity and sensitivity of this technic were compared with those of standard immunofluorescent staining procedures. The unlabeled enzyme method, although a lengthier procedure, has proven to be as specific and at least as sensitive as conventional fluorescent staining. The advantages of the procedure are its permanence, its compatibility with conventional staining procedures, and the avoidance of fluorescent microscopy for evaluation.


Phosphorylase activity in chronic erythremic myelosis. Phosphorylase activity was detected in the cytoplasm of erythroid precursors of 6 of 7 patients with chronic erythremic myelosis (Di Guglielmo syndrome), in proerythroblasts and megaloblasts from 3 patients with pernicious anemia and in 2 patients with severe folate deficiency in neoplastic lymphocytes from 2 patients with acute lymphoblastic leukemia, and in 1 patient with leukemic lymphosarcoma. In all of these patients, most of the erythroid precursors and/or neoplastic lymphocytes contained increased amounts of glycogen when stained with the PAS reagent. Phosphorylase activity was not detected in erythroid precursors obtained from 6 presumed normal individuals or from 3 of 7 patients with a variety of other types of anemia in which the erythroid precursors were PAS-negative. Similarly, phosphorylase activity was absent in lymphocytes obtained from presumed normal individuals. Although the mechanisms responsible for the pathogenesis of PAS positivity are unclear, it is possible that the increased phosphorylase activity found in cells that are PAS-positive may reflect a disorder in the biosynthetic pathway of glycogen.


Nuclear diagnosis of disseminated cancer of the breast. Results of liver-, brain-, and tumor-directed imaging studies. Radionuclide imaging technics to identify metastases in liver and brain have preductive values that show the expected variation depending upon the prevalence of metastatic disease in the population studied. Liver scanning combined with peritoneoscopy may prove more accurate than either study alone for routine use in staging. Brain scanning is most useful when reserved for patients selected because of suspicious neurologic findings or in following the response to therapy of established metastatic disease. "Tumor-directed" scanning agents are useful in certain cases where soft-tissue metastases are suspected but cannot be identified with more conventional procedures.


Seven-drug polychemotherapy in the treatment of advanced and recurrent squamous carcinoma of the female genital tract. Single-agent chemotherapy of advanced and recurrent squamous carcinoma of the female genital tract has been largely ineffective. Combination-drug therapy which has augmented the efficacy of chemotherapy in numerous solid and nonsolid human tumors is usually attended by a degree of toxicity that has discouraged its use against malignancies exhibiting a poor response to single agents. A seven-drug regimen consisting of cyclophosphamide, 5-fluorouracil, actinomycin D, vincristine, cytosine arabinoside, methotrexate, and bleomycin administered during a 24 hour period at 4 week intervals was selected for clinical trial against squamous malignancies of the female genitalia because of its proved broad-spectrum activity among solid tumors and its low incidence of serious toxicity. Severe bone marrow depression occurred during only two of 98 drug cycles involving 23 patients. An objective tumor response was observed in nine of 18 evaluable patients. This regimen appears to be useful in the palliative management of squamous carcinoma of the female genital tract.


The prevention, inhibition, palliation, and treatment of endometriosis. Endometriosis is a variable and enigmatic disease. In this article an attempt has been made to describe principles of management for different groups of patients who have varying manifestations of pelvic endometriosis. Different methods are described for prevention, inhibition, palliation, and treatment of endometriosis.


Prognostic value of antenatal screening. Rates of stillbirths and first-week deaths have fallen steadily in Scotland in the last 30 years. Seventy-five per cent of all such deaths in the City of Dundee now follow the clinical condition of "antepartum hemorrhage," "premature labor," "poor fetal growth," and "lethal deformity," all of which are conditions associated with poorer environmental conditions which are worse in larger industrial cities. Therefore, we are concerned with these clinical situations and with fetal death and subsequent morbidity of children born in those situations. This paper describes in summary a planned study of antenatal screening with assessment of fetal placental hormones, fetal biparietal diameter, and other determinants of fetal well-being, the growth of the child and weight at birth, the quality of the labor, the neonatal state, and the ultimate quality of the child.


Residual urine determination by ultrasonic scanning. In 14 patients with an indwelling catheter, 48 different determinations of bladder volume were made using 5 different ultrasonic methods. The simplest procedure, the largest sagittal diameter of the bladder in the median plane, gave only a rough estimate of the bladder volume. The product of bladder depth, height, and width, as determined from transverse and sagittal scans, showed the best correlation to the true bladder volume (r equals 0.90). Computerized reconstruction of multiple serial cross sections proved less accurate than the much simpler depth, height, width product. None of the methods for quantitation of small volumes was good, but qualitative determination of small volumes was accurate.


Drusen of the iris: in advanced malignant choroidal melanoma. Drusen were found in the iris between the layers of the pigment epithelium and the dilator muscle in an eye of a 52 year old patient that contained a large malignant choroidal melanoma. The significance of this unusual observation in comparison to the common drusen of Bruch's membrane of the choroid is discussed.


Chondroitin sulfate and electron lucent bodies in the pericellular rim about unshrunken hypertrophied chondrocytes of chick long bone. Direct observation of unstained, 1 mm thick blocks of fresh epiphyseal cartilage from tibia of 15- and 18-day-old chick embryos revealed shrunken chondrocytes on its cut surfaces but unshrunken chondrocytes deep within the tissue blocks. The unshrunken hypertrophied chondrocytes are rimmed with refractile substance identified as chondroitin sulfate removable with hyaluronidase. This substance is stained metachromatically red with toluidine blue, and is stained with ruthenium red and with ruthenium red-OsO4. The latter, observed with the electron microscope, is present as an electron dense rim, specifically about the unshrunken, hypertrophied chondrocytes between the plasma membrane and lacunar wall. By rendering the chondroitin sulfate electron dense with RR-OsO4, electron lucent bodies (ELB) were revealed specifically about the hypertrophied chondrocytes. The ELB contain an electron dense core with radiating fibrils. The content and source of ELB, also found in the intercellular matrix, are not known. The 0.1% toluidine blue solution containing 0.2 M MgC12 or 0.4% NaCl or KCl stained juxtanuclear clusters of granules metachromatically red. The location of intracellular granules was believed to represent a cluster of Golgi-derived vesicles. The pericellular metachromatic, RR-OsO4-positive rim is believed to be an accumulation of externalized juxtanuclear metachromatic granules. The possibility that the ELB may also be externalized content of Golgi vesicles was entertained.


Changes in numbers of acidophilic and basophilic connective tissue granulated cells in the intestine of the rat during the estrous cycle. Changes in physiological reproductive states of female Long-Evans hooded rats were directly correlated with changes in the concentration of acidophilic and basophilic connective tissue granulated cells in the lamina propria of the duodenum. Acidophilic granulated cell numbers were significantly higher at estrus and metestrus than at other stages of the estrous cycle or in the immature state. apcidophilic granulated cell numbers during pregnancy were not significantly different from cell numbers quantified during metestrus, diestrus, or proestrus, but were significantly lower than cell numbers during estrus. Late lactation (21-25 days) was associated with significant decreases in the numbers of acidophilic granulated cells from numbers observed during the estrous cycle and during pregnancy. Basophilic granulated cell numbers did not differ significantly during the estrous cycle, although the highest numbers were observed during proestrus. Basophilic granulated cell numbers were significantly higher during pregnancy than during the estrous cycle. Basophilic granulated cell counts in late lactation were comparable to numbers quantified in estrus, metestrus, and diestrus, but were significantly lower than during proestrus and pregnancy.


Agglutinability of cattle red cells. 4. The effect of antiglobulin in comparison with treatment by pronase. The average titre scores varied from zero to 24.7 for the cattle red cells (CRC) from different MZ pairs. These CRC were sensitized with different blood-typing reagents and were titre-tested against the double dilution series of an anti-bovine gamma-globulin serum. A significant negative correlation (r = - 0.82; P less than 0.001) was found between the degree of agglutinability and the amount of neuraminic acid of the surface component of CRC cleaved by pronase. After pronase treatment of the CRC it could be demonstrated that (1) the activity of the V and E'3 blood factors became destroyed; (2) three new specific receptors became evolved; (3) the degree of 'direct' agglutinability due to the A2, O3, W, S2 and Z anti-sera did not parallel with the titre scores obtained in the anti-globulin tests.


[Radioimmunologic assay of alpha-fetoprotein in various normal and pathological conditions]. AFP can be measured by radioimmunoassay in 52 hours using an incubation at 18 degrees C for 48 hours and the double antibody solid phase method to separate the free labelled AFP from the labelled AFP bound to the antibodies. In these conditions, the sensitivity and precision of the assay are very satisfactory and the results are directly in correlation with the results obtained using the classical double antibody method. In the serum of normal people, except pregnant women, the concentration of AFP is not detectable or is less than 20 ng/ml. In the serum of pregnant women, AFP increases from the 8th. week of gestation to the 32nd., then stabilises or gradually decreases.


The role of alpha1-antitrypsin and alpha-macroglobulins in the uptake of proteinase by rabbit alveolar macrophages. Proteinases in plasma bind largely to alpha1-antitrypsin and alpha-macroglobin. The latter is represented in humans by alpha2-macroglobulin and in the rabbit by alpha2-macroglobulin and alpha1-macroglobulin. There is a selective and rapid uptake of proteinase-macroglobulin complexes by rabbit alveolar macrophages. It was found that complexes of labelled trypsin or subtilopeptidase A with rabbit alpha1-antitrypsin do not become similarly associated with rabbit alveolar macrophages. Moreover, proteinase-alpha1-antitrypsin complexes failed to inhibit the uptake of labelled proteinase-macroglobulin complexes. Thus, the interaction with macrophages of proteinase-macroglobulin complexes represents a pathway of proteinase metabolism distinct from that involving proteinase bound to alpha1-antitrypsin.


Ascorbic acid requirements and metabolism in relation to organochlorine pesticides. Those organochlorine pesticides which possess both high lipoid solubility and high resistance to biodegradation are prone to accumulation in animal tissues and produce relatively long-term effects as toxicants. Such compounds, typified by DDT, Dieldrin, and Lindane, are profound inducers of hepatic microsomal enzymes, including parts of the glucuronic acid and ascorbic acid biosynthetic pathways. Consequently, administering such pesticides to rats in accompanied by enhanced formation and excretion of D-glucuronic acid and L-ascorbic acid, or D-glucaric acid in the case of guinea pigs. Secondarily, the efficiency in biodegrading the pesticides is reduced in ascorbic-acid-deficient guinea pigs with correspondingly greater residue accumulation in tissue. This would aggravate chronic toxic effects of the compounds. Finally, the capacity of the liver to adapt to the presence of such toxicants through enhanced microsomal enzymatic levels appears to be sensitive to its ascorbate status. Impaired enzyme induction is apparent quite early during ascorbic acid depletion in guinea pigs. The enhanced turnover of ascorbate produced by such pesticides, the poor enzymatic adaptation to them during ascorbate depletion and the dependency of the oxidase system upon adequate ascorbate, all point to the central significance of ascorbate status in the liver, and possibly other tissues, as a determinant of their chronic toxicity.


Immunological and chemical correlation between alpha-fetoproteins from human and several mammalian species. Alpha-Fetoproteins of several animals were purified and their molecular weights, amino acid compositions and peptide maps were compared, demonstrating the close similarities. These data indicated that the alpha-fetoproteins of mammalian species have closely related antigenical and chemical structures. Rabbits and horses were immunized with human alpha-fetoprotein, and it was observed that the animals produced antibodies reaction not only with human alpha-fetoprotein but with their homologous alpha-fetoproteins. The results were interpreted as the breakdown of the tolerance to their own alpha-fetoprotein.


Alpha-Fetoprotein levels in normal males from seven ethnic groups with different hepatocellular carcinoma risks. Alpha-Fetoprotein (AFP) levels of 1,335 males (15 years and older) of seven ethnic groups (Chinese, Indians, and Malays from Singapore, Caucasians from Lyon, and Blacks from Nairobi, forest, and the savanna region of the Ivory Coast) were determined by radioimmunoassay. A few elevated levels (up to 30 nanounits/ml) were detected in some normal individuals, especially in the older age-groups. In addition, there was a systematic age-dependency of AFP levels particularly evident in the groups from Singapore-Lyon, in which there was a 50% AFP increase between the ages of 20 and 40. Comparison between Africans on the one hand and people from Singapore-Lyon on the other hand revealed highly significant differences (p less than 0.001), especially in the younger groups, whereas Chinese, Malays, and Indians from Singapore had very similar AFP pattern; this suggests an important role for environmental factors in the regulation of AFP levels. The age dependency of the presumed effect of environmental factors is in keeping with experimental data showing that young animals respond more vigorously to AFP-stimulating factors. Although the incidence of hepatocellular carcinoma (HCC) differs in the three Singapore groups (the highest in Chinese and the lowest in Indians), no relationship was observed in this study between mean AFP level and HCC incidence in Singapore.


Alpha-Fetoprotein and hepatitis B antigen in hepatocarcinogenesis. Blood from 394 unselected autopsy cases was examined for HB Ag, HB Ab and AFP. Liver morphology of 71 cases of cirrhosis with hepatoma and 32 cases of cirrhosis without hepatoma was studied in detail and correlated to HB Ag, HB Ab, and AFP. Significantly lowered humoral immunity to HB Ag exposure was established for the cirrhosis with hepatoma cases. The exposure rate for both cirrhosis cases with hepatoma and cirrhosis cases without hepatoma was the same (48%), but about 80% of each exposed group were either HB Ag or HB Ab positive. The cirrhosis with hepatoma group tended to be HB Ag positive and the cirrhosis without hepatoma group tended to be HB Ab positive. The lowered immune response seems to be specific to the hepatoma association, because the group with neoplasms other than the hepatoma reacted exactly the same as the group of the cirrhosis without hepatoma. Twenty-five per cent of the cirrhosis with hepatoma were associated with inactive cirrhosis and 75% were associated with active cirrhosis. Seventy-two per cent of the inactive cirrhosis cases with hepatoma were exposed to HB Ag, but only 42% of the active cirrhosis cases with hepatoma were exposed to HB Ag. On the morphological basis, the inactive cirrhosis was interpreted as an impaired cellular immunity and the active cirrhosis as a delayed hypersensitivity reaction. The possibility was discussed that both are important factors in the development of hepatoma preceded by cirrhosis. AFP tends to be positive in the inactive cirrhosis with hepatoma as well as HB Ag, but the relationship between AFP and HB Ag for hepatocarcinogenesis needs further investigation.


Early detection of hepatoma: prospective study in liver cirrhosis using passive hemagglutination and the radioimmunoassay. (1) Passive hemagglutination and radioimmunoassay are suitable methods for the detection of AFP in the low concentration range. (2) In 3.72% of the cases a clinically unknown carcinoma was found in an unselected group of patients with liver cirrhosis. (3) 21.9% of the patients showed AFP elevations up to 2000 ng/ml. In 10.6% of this group, increasing titers demonstrated a primary liver cell carcinoma. In 89.4% a transitory rise of AFP was not associated with tumor growth. Levels return to normal values within three months in 90% of the cases. (4) Transitory AFP elevations are not correlated to clinical conditions (praecoma, coma, delirium, bleeding, ascites, shunt) or to biochemical parameters (GOT, GPT, bilirubin, prothrombin complex time, gamma-globulin). (5) A temporary rise in AFP is more frequently observed in groups with high hepatoma incidence than in groups with low hepatoma incidence. (6) Therefore, it may be suggested that a transitory rise of AFP could reflect a "primary reaction" of carcinogenesis. (7) Primary liver cell carcinoma is found to be more frequent in posthepatitic than in postalcoholic, cryptogenic, and other cirrhosis and to be more frequent in australia-antigen positive than in australia-antigen negative cases. (8) Routine serological tumor antigen screening of patients with a precancerous disease is useful.


Immunoelectronmicroscopic study of alpha-fetoprotein synthesis in hepatoma cells. In the hepatoma cells, AFP synthesis was found to occur through ribosomes of the rough endoplasmic reticulum, since AFP was demonstrated around ribosomes of the rough endoplasmic reticulum by the peroxidase antibody technique. The secretory process was suggested to be as follows: smooth endoplasmic reticulum takes a part and the Golgi apparatus does not. Concerning the early transitory appearance of AFP in the course of hepatocarcinogenesis by 3'Me-DAB, AFP might be produced by proliferated ampulla cells, which exist between the cholangioles and liver cell cords.


Synthesis of alpha-fetoprotein by rat ascites hepatoma cells. AH-66 rat ascites hepatoma cells incorporated [14C]leucine into the AFP fraction. In a cell-free system, hepatoma ribosomes were found to be active in AFP synthesis whereas the supernatant fraction from hepatoma had no specific effect on AFP production. The amount of [14C]leucine incorporated in AFP by membrane-bound polysomes was 20 to 90 times higher than that by free polysomes, suggesting that AFP is mainly synthesized on membrane-bound polysomes. DBcAMP inhibited the growth of hepatoma cells. However, the incorporation of [14C]leucine into the AFP fraction as well as into total proteins was stimulated by DBcAMP.


Alpha-Fetoprotein and albumin synthesis during the cell cycle. The synthesis of alpha-fetoprotein and albumin in two clones of AH66 hepatoma was studied. (1) Amounts of AFP and albumin synthesized by the C-4 clone were 2.9 and 0.28 X 10(-7) mug per cell per hour, respectively. AFP and albumin amounts synthesized by the A-1 clone were 2.5 and 1.8 X 10(-7) mug per cell per hour, respectively. (2) The cell cycles of the C-4 and A-1 clones were as follows: C-4 clone: mean generation time 20.5 hours: G1, 10 hours; S, 7 hours; G2, 4 hours; and M, 30 minutes. A-1 clone: mean generation time 50.7 hours: G1, 36 hours; S, 10 hours; G2, 4 hours; and M, 30-60 minutes. (3) AFP was found to be synthesized from late G1 phase to the end of the S phase for 9 hours in C-4 and 25 hours in A-1. The albumin production was from late S phase to the beginning of the G2 phase for 4 hours in C-4 and 9 hours in A-1, which were approximately half or one-third of the time spent on AFP production. (4) The double staining with fluorescent-conjugated antibodies to AFP and albumin demonstrated that AFP and albumin are probably synthesized by different cells.


Clinical significance of alpha2H-globulin. Incidences of alpha2H were investigated in sera from patients with various malignant and nonmalignant diseases. With counterimmunoelectrophoresis, alpha2H was detected in a wide variety of neoplastic diseases, and also in nonmalignant hematological disorders. Diagnostic significance was described regarding primary hepatoma and acute myelogenous leukemia. Factors affecting alpha2H levels in sera were also discussed in relation to the neoplastic and nonneoplastic disease process.


Relationship of the biosynthesis of alpha-fetoprotein, albumin, hemopexin, and haptoglobin to the growth state of fetal rat hepatocyte cultures. AFP and albumin are produced by arginine-synthesizing fetal rat hepatocytes in vitro. AFP and hemopexin production are coupled to hepatocellular proliferation, whereas albumin and haptoglobin production are not. During the cell cycle, AFP is synthesized prior to S and released prior to M. AFP may play a role in regulation of hepatocellular growth through estradiol binding and modulation of the intracellular concentration of lipoprotein (VLDL).


Microheterogeneity of rat alpha-fetoprotein. A purified and homogeneous preparation of rat AFP, as judged by both electrophoresis on Cellogel and immunoelectrophoresis, was separated into two components, AFPa and AFPb, by disc electrophoresis on 7% polyacrylamide gel. These two components had a definite difference in electrostatic net charge and gave only a single band on SDS-electrophoresis. Immunological reactivity or electrophoretic separation or mobility of the two components could be altered by treatment with either sulfhydryl inhibitors or reducing agents but not by treatment with protein denaturants. Electrophoresis of neuraminidase-treated AFP on 5% polyacrylamide gel yielded clearly separable, slower moving four to six and finally two components depending on the time of incubation with neuraminidase. The time-dependent conversion of faster into slower migrating components of both AFPa and AFPb upon neuraminidase treatment was confirmed by reelectrophoresis of separated and similarly treated AFPa and AFPb. Two bands of sialized or desialized AFP were also observed on isoelectric focusing. Both AFPa and AFPb treated with and without neuraminidase gave single fused precipitin lines against the antiserum in Ouchterlony double-diffusion analysis. On the basis of the changes in electrophoretic mobilities of the intermediates following neuraminidase treatment, AFPa and AFPb were estimated to have at least 2.5 and 4.5 molecules of sialic acid per molecule, respectively.


Diagnosis of systemic lupus erythematosus. Importance of antinuclear antibody titers and peripheral staining patterns. Titers and patterns of antinuclear antibodies (ANA) in sera from 134 normal blood donors, 20 patients with rheumatoid arthritis, 15 patients with systemic scleroderma, and 32 patients with diagnosed or suspected systemic lupus erythematosus (SLE) were studied. The difference between the findings with sera of patients with SLE  and normal subjects in terms of high (greater than 160) titers of ANA was greater than in terms of peripheral staining patterns. However, in comparing sera from patients with SLE with sera from patients with other connective tissue diseases, greater differences were found in the incidence of peripheral patterns of ANA compared to differences in the frequency of high ANA titers. Maximum specificity in the diagnosis of SLE was achieved when both titers and patterns of ANA were considered.


Ventricular arrhythmias during unstable angina pectoris. In order to study the occurrence and frequency of ischemia-induced ventricular arrhythmias, we analyzed 105 episodes of spontaneous angina pectoris occurring at rest in 28 hospitalized patients with unstable angina pectoris and proved coronary artery disease. Of 24 patients with serious ventricular arrhythmias during pain, 17 (57%) were arrhythmia-free during monitoring. In the other four patients, 17 of 29 (59%) pain episodes were associated with serious ventricular arrhythmias, and three of these four had serious ventricular arrhythmias during pain-free periods. Each patient tended to manifest the same type of arrhythmia during repeat episodes of pain. It appears that continuous electrocardiogram (ECG) monitoring is important during the initial hospitalization of the patient with unstable angina. The presence of ventricular arrhythmias during pain-free periods indicates a high risk for serious ventricular arrhythmias during episodes of spontaneous pain. These patients should be considered for continued ECG monitoring and antiarrhythmic therapy.


Effects of adrenaline, angiotensin and calcium on spontaneously active and potassium-depolarized rat portal veins. 1) Adrenaline and angiotensin increased both frequency and developed tension of spontaneously active rat isolated portal veins. Adrenaline 10(-7) M and angiotensin 10(-9) M, produced a maximal increment of the amplitude of twitch contraction. Greater concentrations elicited a tetanic state, which in all cases had the same amplitude as the maximal effects obtained on single twitches. This suggests that there exists a dissociation between membrane and electrical phenomena, so that the highest intensity of the contractile system activity is attained at agonist concentrations lower than those which result in maximal decrease of membrane stability. (2) Increasing K to a 106 mM final concentration caused a persistent contracture. Adrenaline and angiotensin exhibited pharmaco-mechanical properties, as later addition of both substances produced an increase of the contracture tension. (3) Ca dose-response curves were performed on previously Ca-depleted veins, after the addition of K 106 mM, K-plus-adrenaline or K-plus-angiotensin. Both substances made the Ca responses rise steeply, as evidenced by the leftward shift of ED50. This provides support to the belief that pharmacomechanical coupling is mediated through an increase of the membrane permeability to Ca.


Rapid eye movement (rem) sleep deprivation: effect on acid mucopolysaccharides in rat brain. The effect of rapid eye movement (REM) sleep deprivation on the total content and proportion of different mucopolysaccharides (AMPS) containing uronic acid in rat brain was studied. REM sleep deprivation was induced by the water tank methods. Five experimental groups of animals were used: control, stressed, REM sleep deprived, post-stress sleeping and post-deprivation sleeping rats. No changes of AMPS were observed in any of the experimental groups when the whole brain was analysed. A significant increase of AMPS was found in the cerebral hemispheres of stressed and REM deprived rats. A significant decrease of AMPS was observed in the cerebellum and brain stem. A further increase of AMPS was found in the cerebral hemispheres after the rebound of REM sleep following its deprivation, and after the recovery sleep following the stress. A significant increase of AMPS was found in the brain stem of rats allowed to recuperate after REM deprivation or stress as compared with the stressed and REM deprived animals. Recovery sleep induced a significant increase of AMPS in the cerebellum in previously stressed rats, while previously REM deprived rats exhibited a further decrease of AMPS from control values. The possible functional meaning of these results is discussed in relation to the role of REM sleep in protein synthesis and learning and memory processes. Intriguing, well-controlled positive findings and the fact that no experimental design is known where stress is minimal while REM deprivation is 100 per cent, justify and encourage continued efforts in studying the biochemical state of the brain during sleep and/or its alterations.


Alpha-Glycerophosphate dehydrogenase (insoluble) and lactic dehydrogenase activities in the skeletal muscles of two insects. The flight muscle preparations of the dragonfly Pantala flavescens and the aquatic beetle Cybister confusus showed extremely low levels of lactic dehydrogenase activity and high levels of alpha-glycerophosphate dehydrogenase (insoluble) activity. The activities of these two enzymes in the leg muscle of the beetle were approximately the same (1:1), but lactic dehydrogenase activity was several times higher than that in the flight muscles of both Insects. These results have been interpreted as indicating the high energy-yielding demands of the flight muscles during continuous sustained activity, while the leg muscles of the beetle which are involved in swimming activity derive their energy predominantly through anaerobic glycolysis.


The effect of secretin on pentagastrin-stimulated secretion of gastric pepsin and acid in rats. Rats with chronic gastric fistulas were stimulated for 12 or 24 h with constant intravenous infusion of pentagastrin. When secretin was also infused for the last half period of the experiment, respectively, 6 or 12 h, the volume of gastric secretion and HCl output were significantly reduced but the concentration of pepsin was significantly increased. The dissociated effect of secretin on gastric acid and pepsin secretion reported previously in man, dog and cat was also found in the rat.


Replacement time for alveolar lipid removed by pulmonary lavage: effects of multiple lavage on lung lipids. Daily washing in vivo of the lung with 0.15 M saline did not deplete the Beagle dog lung of surfactant lipids, but rather increased the quantity of surfactant lipid in the tissue. Replacement time for the lung lipids removed by the lavage was approximately 5 hours. This rate is one indication of the time required for movement of surfactant lipid from storage areas to the surface of the alveoli. The increase in tissue surfactant lipid following multiple lavage suggests that the rate of surfactant lipid synthesis is controlled in part by the level of surfactant lipid in the alveoli.


Effect of coumarin and some coumarin derivatives on active transport and passive diffusion of sugars by chicken and rat intestine, in vitro. The effect of coumarin, 4-hydroxycoumarin, coumarin-3-carboxylic acid and acenocoumarol on the active transport of D-galactose and the passive diffusion of arabinose by intestinal sacs was studied. All these substances, when added to the mucosal medium of incubation at concentrations from 10(-4) to 10(-3) M, inhibit the active transport of D-galactose and increase the diffusion of arabinose. Oxygen uptake by the intestinal tissue is only inhibited by coumarin-3-carboxylic acid. The results suggest that the effects obtained are probably due to an alteration in intestinal permeability, to inhibition of cell metabolism and to molecular size.


Rate of incorporation of dietary fat in gold thioglucose obesity in mice. Gold Thioglucose injections in mice are followed by a rapid accumulation of fat in the carcasses. The incorporation of an oral dose of [3H] glyceroyl tripalmitate in body fat stores showed after GTG-treatment a transient but significant increase and a return to normal values within 6 weeks. The rate of incorporation of dietary fat into the body was estimated from these values as well as from food intake and fat content of the diet (2.5 per cent). The resulting curve showed great similarity with the first differential of the curve of total body fat accumulated during that period. The rate of incorporation of dietary fat into body stores is apparently modified in GTG obesity in mice.


[Effect of 5-HT on insect cardiac activity in vivo]. Injection of the Insect Locusta migratoria with 10 mul of normal saline produces an important (10 per cent) but short (1 min) increase of the cardiac rhythm. Adding various concentrations of 5-HT with the saline always results during the following ten minutes in a greater increase of the cardiac rhythm than the controls.


Beneficial effects of L-methionine in choline deficient diets of Oryzaephilus mercator (Fauvel) (Coleoptera). Choline is an indispensable vitamin for Oryzaephilus mercator as its ommission in the diet results in slow growth and poor survival. These deficiency symptoms can be overcome by adding L-methionine to the choline-deficient diet. L-methionine however can improve the efficiency of the diet only up to a certain concentration beyond which no beneficial effects are recorded.


Effect of thermal stress and dehydration on plasma levels of glucose, free fatty acids and growth hormone in the pigeon. Pigeons were subjected to high ambient temperature and water deprivation for 3 days (28 degrees C, 31 degrees C and 36.5 degrees C respectively). After 3 days of heat stress and dehydration, the plasma levels of glucose, free fatty acids (FFA) and growth hormone (GH) were measured. Although the level of plasma glucose was not significantly altered, FFA and GH were found to be significantly increased. The possible mediation of the neurohypophysial hormone, vasotocin, in the syndhronous rise in plasma GH and FFA, is suggested.


Vascular and noradrenalic reactions in the musculocutaneous bed during hypothalamic stimulation. When electrical stimulation is applied to the ventromedial hypothalamic zone one observes an increase in systemic blood pressure. There also occur blood pressure variations in the isolated femoral circuit: two distinct phenomena were observed. The early event, being either an increase or a decrease in peripheral resistance, is directly related to the amount of noradrenaline produced locally. The late event is due to catecholamines arriving from the general circulation. Inhibition of local catecholamine release through the baroreceptor reflex and inhibition of ganglionic transmission by a large and sudden increase in adrenaline blood levels do influence the response in the isolated femoral circuit. Moreover the peripheral vasomotor tonus seems to be influenced by yet another mechanism, independent of local catecholamine release. This delicate mechanism depends on the balance between the degree of excitation of hypothalamic pressor (medial) and depressor (lateral) zones.


[Hypothalamo-hypophysial-thymic axis in the rat]. Thymic extracts, with proved immunologic properties, decrease, in vitro, RNA and DNA synthesis of hypothalamic fragments of adult rat and have no effect on hypophysis culture; we found a similar action in vivo. Growth hormone inhibits the nucleic acids synthesis of thymic fragments; no action is found with hypothalamic extracts. Hypothesis, that thymic activity is dependent on hypophysis with retroaction on hypothalamus, is discussed.


Effect of PCO2 on epinephrine-induced lipolysis in isolated fat cells. The effect of different pHs obtained by changing the PCO2 and the effect of PCO2 at constant pH on the lipolysis induced by epinephrine in isolated fat cells have been investigated. An inhibition of activated lipolysis was found in acidosis while in alkalosis no significant change was detected. When the experiments were performed at different PCO2s but at constant pH, the results showed an inhibition of lipolysis by high PCO2 whereas low PCO2 did not affect it. It is concluded that either acidosis or high PCO2 lead to an inhibition of the lipolysis induced by epinephrine in isolated fat cells. As regards alkalosis and low PCO2 it seems likely that the intracellular pH is not affected to the same extent as in alkalosis by high [HCO(-3)] or under the conditions of the present experiments the [H+] needed to alterate lipolysis was not reached.


Ionic regulation of the haemolymph in the larvae of the dragonfly Aeshna cyanea (Müller) (Odonata, Anisoptera). The ionic regulation of the haemolymph of larvae of Aeshna cyanea (Müller) was studied by means of two types of experiments. In the first the change in internal ionic composition was followed as a function of the time spent in a given experimental medium. These experiments led to the conclusion that: 1. the haemolymph composition does not change when larvae are starved in tap water for 10 days; 2. the haemolymph ionic concentrations (Na and Cl) have initial marked increase when the animals are kept in hypertonic media of diluted sea water; after 80 hours however, both concentrations stay constant. In a second series of experiments the internal ionic concentration was compared to a series of different concentrations of external media. From this, the relation between the internal and external ionic concentration was elaborated : in hypotonic media the internal Na and Cl concentrations stay constant, in hypertonic media there is a parallelism between the increase of the external concentration and haemolymph concentration, the internal Na concentration being always slightly hypertonic, the Cl concentration markedly hypotonic. Finally, when larvae are placed in glass distilled water, the internal Na and Cl concentrations begin to decrease after 60 hours.


Quantitative measurement of the influence of plasma sodium concentration on the overall tubular reabsorption of sodium. Comparative experiments on isolated dog kidneys perfused with heparinized blood with or without dilution of the blood by isotonic or hypotonic saline demonstrated that the fractional excretion of sodium is modulated positively by plasma sodium concentration. This relationship was evaluated quantitatively and corresponded to the values found in the whole animal. The renal response to the variations of plasma sodium concentration is therefore autonomous and its mediation by extrarenal natriuretic or antinatriuretic factors cannot be demonstrated in the narcotized animal.


[Attenuation and smoothness of injected signals in a Rijlant network]. The V.C.G. methods must handle the E.K.G. information through weighing systems, because of the volume and the eccentric location of the heart in the chest, of the inhomogeneity of the surrounding tissues and of the irregular shape of the body surface. Most of these weighing systems use compensating networks. On the other hand, Rijlant simulates an homogeneous medium where the cardiac currents spread and where remote observation would eliminate much of the distorsions. In order to understand how such a simulated medium operates, we drive it by means of sinusoidal waves and we observe how the amplitude and the phase of the signals change when we switch on the different leads. Each value recorded at a same level in the network may be considered as the sum of a true proximity effect added to an other component depending upon the lead orientation. The latter is referred to a mixed result of the driving waves that we call "the modulus effect". From the injection level to the other levels, the proximity effect rapidly vanishes while the modulus effect does not disappear until grounded level is reached. The decrease of the modulus effect either to zero (grounded level) or to a finite value (free end of the network) is not expressed as a formula because Rijlant's network does not simulate an euclidian space. We conclude that, at least as the proximity effect is concerned, the instrument performs what it is expected to do.


[Influence of hypo- and hyperthyroidism on the turnover rate of noradrenaline, dopamine and serotonin in various rat cerebral structures]. The effect of chronic treatment with tyroxine (T4) or propylthiouracile (PTU) on the turnover of norepinephrine (NE), dopamine (DA) and 5-hydroxytryptamine (5-HT) has been studied in various areas of the rat brain (brain stem, hypothalamus, striatum and "rest of the brain"). The turnover of NE and DA was determined by the decay in endogenous levels after inhibition of tyrosine hydroxylase by alpha-methylparatyrosine and the turnover of 5-HT was evaluated by the initial accumulation of endogenous 5-HT after inhibition of monoamine oxydase by pargyline. T4 treatment accelerated the release of DA from the striatum but had no significant effects on NA release in the various cerebral areas : nevertheless the NE endogenous level was significantly reduced in the brain stem. PTU treatment delayed the release of DA and NA only from the "rest of the brain". Concerning 5-HT, the only significant variation was observed in the hypothalamus of PTU-treated rats and implied increased turnover. The possible relations between the changes in cerebral monoamines turnover and the behavioural alterations which are observed in thyroid disfunction are discussed.


[Free amino acids of hemolymph during pubertal molting and senescence in Spaeroma serratum (Isopoda, Flabellifera]. In Sphaeroma serratum, the amino-acidemia is high (about 120 mg/100 ml) during the intermolt stages. The relative proportions of the different free amino acids show only slight differences in young males and senescent ones. At the time of the puberty molt, the total amino-acids concentration (including taurine) of the hemolymph increases sharply during premolt (up to 267 mg/100 ml), falls after ecdysis (down to 97 mg/100 ml and then rises up to the level of the intermolt stage. These variations are due not only to concentration or dilution of the whole hemolymph constituents, but also to specific modifications of the concentration of some of the free amino-acids with respect to the others. These variations of the amino-acidemia, mainly those of serine, proline, glycine, alanine and of the amine taurine, could play a definite role in increasing the osmotic pressure of the hemolymph during premolt, as a preparation to ecdysis.


Localization of the acetylcholine receptors in denervated muscles of rats. ACh contractures of denervated lumbricalis muscles of rats were washed in Tris-methanesulfonate solutions and in sucrose solutions. The peak tension was diminished by about 80% following a 30 min exposure to solutions containing 400-600 mM glycerol when Tris solutions were used in the testing period, and by about 50% when sucrose solutions were used. The amplitude of the membrane potential changes provoked by ACh was decreased by about 36% following the glycerol treatment. The treatment had no effect on the ACh-induced 45Ca uptake of muscles. Electron microscopy of the glycerol-treated muscles showed widespread vacuolization apparently originating from swelling and disruption of the T system. It was concluded that ACh receptors which arise in the muscle membrane following nerve section are distributed in the external surface membrane and in the membranes of the T system.


Changes in metabolite concentration in the plasma, liver and muscle of feed restricted Japanese quail exposed to cold. Quail fed ad libitum and 50% ad libitum were cold exposed for several weeks, during time control quail remained at 21 degrees C. The concentration of plasma glucose, FFA, and uric acid, tissue glycogen and carcass fat content was measured at the end of the cold exposure period. Quail fed ad libitum showed no significant change in the levels of plasma and tissue metabolites, or the carcass fat content, following cold exposure. The feed consumption by the cold exposed quail increased, and the mean body weight showed little variation from that of the controls. Feed restricted quail which were cold exposed lost significantly more weight, and had a lower ranked fat content than their controls. Whereas feed restriction caused a lowering of the liver glycogen concentration in both treatment groups, muscle glycogen levels were higher than in quail fed ad libitum. However, cold exposure was not accompanied by a change in muscle and liver glycogen levels in feed restricted quail. Feed restricted quail at 21 degrees C were hypoglycaemic and hyperlipaemic compared to quail fed ad libitum, but cold exposed feed restricted quail had a much higher plasma glucose concentration than the controls. The ranked carcass fat content was inversely related to plasma FFA level in both control and cold exposed feed restricted quail. It is suggested that both a glycolytic and lipid mobilizing response to cold is obtained in quail whose body reserves are not spared from catabolism by adequate dietary nutrient absorption, and the possibility of gluconeogenesis from precursors produced by proteolysis is indicated.


Studies on human urinary arylamidases.

Studies on human urinary arylamidases. Human urinary protein was found to contain enzymes that hydrolyze leucyl-, alanyl-, and glycyl-prolyl-beta-naphthylamides. The kinetic constants of these enzymes were determined and their chemical properties studied. The values for Km calculated for leucyl-, alanyl-, and glycyl-prolyl arylamidases were 3.7 times 10(-5) M, 7.1 times 10(-5) M, and 1 times 10(-4) M, respectively. The pH optima for the hydrolysis of Leu-beta-naphthylamide and Ala-beta-naphthylamide were between 7.4-7.8; whereas for glycyl-prolyl-beta-naphthylamide, it was around 8.8. Unlike the glycyl-prolyl-arylamidase which was inhibited by Co2+ and Mn2+, the other two arylamidases were slightly activated by Co2+. p-Chloromercuriphenyl-sulfonate and puromycin significantly inhibited leucyl-, and alanyl arylamidases. The mean values for 24-h urinary output for leucyl-, alanyl-, and glycyl-prolyl arylamidases in normal human male subjects were 4.32, 9.97, and 2.2 units, respectively.


[Effects of a high-fructose diet on various characteristics of metabolism in rat small intestine]. During the fructose intestinal transfer in vitro, in the rats receiving a normal diet, no glucose is synthetized. In the high-fructose diet rats, glucose appears in the intestinal wall and in the serosal fluid. This synthesis is probably realized from fructose. When fructose is used as substrate, the intestinal tissue of these latter animals shows a greater oxygen consumption than that of the former animals.


[Hepatic lipids of intact or thyroidectomized rats after administration of tetracycline or ethanol]. The purpose of this work was to study the quantitative modifications of the hepatic lipids in adult thyroidectomized rats after administration of tetracycline or ethanol (acute dose or prolonged ingestion). 1. - Thyroidectomy did not inhibit the accumulation of fat in the liver of fed euthyroid or hypothyroid rats after intraperitoneal infusion of tetracycline (320 mg/body weight in 2 injections at an interval of 16 h, the diet containing 6% of lipids). 2. - Sixteen hours after the oral administration of a single large dose of ethanol (5 g/kg body weight), there were only found some small modifications of the lipid composition of the liver in fasting euthyroid or thyroidectomized rats, receiving a diet with 6% of lipids before the experiment; on the contrary, when the diet contained 19% of lipids, a fatty liver occurred in the intact rat, but not in the thyroidectomized rat. 3. - The prolonged ethanol intake (in a 20% solution in water) for 5 months with a diet containing 19% of lipids did not induce a fatty liver in intact rats but produced a decrease of hepatic non-phosphorus lipid and an increase of the cholesterol amounts. After the administration of L-thyroxin (10 mug/100 g body weight per day) to these alcoholic thyroidectomized rats during 2 weeks, it was found an increase of the hepatic non phosphorus lipids till an higher amount than in the euthyroid rats. 4. - The hepatic phospholipid amounts were relatively constant in the different experiments. These results accounting for this differential effects were discussed.


A method for studying the peripheral mediators of the dental pain induced by electrical stimulation. A useful method is described in this paper for studying the mediators released from tooth pulp of the dog during the electrical stimulation of dentine. This method is based upon the perfusion of the pulp and superfusing the return continuously over the isolated, in cascade, cat jejunum and rat stomach fundus strip. The presented evidences in this study indicate the possibility of the release of bradykinin and prostaglandin - like materials from the tooth pulp during the electrical stimulation of dentine. Possible relationship of these mediators and tooth pain due to the electrical stimulation of dentine is discussed.


On the sterols of some ascidians.

On the sterols of some ascidians. The lipid content of sea squirts is low, namely less than a half percent of the fresh weight. Lipids consist of about seventy percent of saponifiable lipids and of about twenty percent of non-saponifiable lipids. Both types of these lipids, including sterols, can be synthesized from acetate by these animals. Small amounts of C30 sterols were observed only in Microcosmus sulcatus and Halocynthia papillosa, the species with a low content of C27 sterols and a high content of C28 sterols. In addition these species contained considerable higher amounts of sterols with a double bond at the C22 position than Ciona intestinalis and Styela plicata did.


[Action of isopropanol on rat lipid metabolism: complementary studies on mechanisms implicated in hepatic accumulation of triacylglycerides]. Isopropanol administration (3 g/kg, p.o.) determines in the rat liver an inhibition of fatty acid oxidation, an enhancement of fatty acid esterification into triacylglycerols as well as an inhibition of lipoprotein secretion which appears to be related to alterations in hepatic phospholipids and which differentiates isopropanol from ethanol induced fatty liver. Disturbances in peripheral lipolysis following isopropanol administration are found only in mature rats and are not necessary to the fatty liver induction.


Interaction between lindane and micorbes in soils. Three lindane (gamma-1,2,3,4,5,6-hexachlorocyclohexane) treated soils were studied under laboratory conditions to determine the interaction between lindane and the soil microorganisms. Microbial populations and respiration were monitored to study insecticide effects. Formation of lindane degradation products and chloride content were examined to determine effects of the microorganisms. Some populations in lindane treated soils showed temporary declines but all ultimately recovered to at least the level of the controls in 16 weeks. Respiration was stimulated over a 9-week period especially in the sandy and clay loams, suggesting the possibility of microbial degradation of the insecticide. Lindane degradation products separated and identified by TLC included gamma-2,3,4,5,6-pentachloro-1-cyclohexene (gamma-PCCH), gamma-3,4,5,6,-tetrachloro-1-cyclohexene (gamma-TCCH), gamma-3,4,5,6-tetrachloro-1-cyclohexene (gamma-TCCH), and pentachlorobenzene (PCB). Chloride production increased in soils treated with higher levels of lindane.


[Thymus changes in chicks following treatment with small doses of insecticides]. Effects of small doses of insecticides (Fenclorfos and Heptaclor) upon the thymus of chickens were followed. Treatment began at the age of 3 weeks and continued during 4 or 8 weeks. Doses used were: 1 ppm for Heptaclor and 1 ppm and 0.5 ppm for Fenclorfos. Total nucleic acid, total protein, RNA, DNA, and amino acid nitrogen contents were determined, as well as GOT and GPT activities and weight of the organ. Results are interpreted as being due to the action of insecticides on the hypothalamus-hypophysis-adrenals axis. They depend on the nature and dosis of insecticide, as well as on the duration of the treatment.


[Study of Y-chromatin in the buccal epithelium of man]. Investigation was carried on by fluorescent-microscopic technique, smears of buccal epithelium being stained with acrichinium-mustard gas. The extent, form, location in a nucleus and the frequency of Y-chromatin in human buccal smears were studied. In medico-genetic practice this method is employed for initial determination of chromosome sex in disease associated with anomalies of sex chromosomes.


[Improvement of the method of staining cationic proteins of the lysosomes of polymorphonuclear leukocytes in laboratory animals and man]. Some methodological drawbacks of the previously offered by the author method for speedy staining of lysosomal cation proteins have been eliminated. The method is shown to be applicable for histochemical staining of lysosome cation proteins of human polymorphonuclear leucocytes. Staining is carried out on blood smears and preparations-impressions from the surface of incision on the organ under study. The improved method for staining of cation proteins can be used in determining the fate of cytophagous bacteria, in cytochemistry of cation proteins of polymorphonuclear leucocytes, in detecting vital and perished bacteria in inflammation foci in bacterial infectious processes.


[Opportunities for the study of disorders of the microcirculation in the heart in different variants of ischemic disease]. To ascertain the occurrence of disorders of blood circulation in the heart, in particular, in acute impairment of the coronary blood circulation and repeated infarctions, perfusion of the vascular system of the heart with a solution of a stain was carried out. It was shown that those parts of the heart where, due to certain reasons, there were created conditions for development of impairment of the microcirculation, remained unstained. It was corroborated by the results of histological and cytological investigations of the myocardium. The coincidence of clinical observations with the results of the perfusion may indicate that this method of investigation of the heart at autopsy makes it possible to understand better prevital disorders of capillary blood circulation. The method of perfusion of the heart with a stain is set forth in the work in detail.


Experimental thiamine deficiency. Neuropathic and mitochondrial changes induced in rat muscle. Whether pure thiamine deficiency produces a neuropathy in Mammalia is still debated. Rats were pair-fed-synthetic diets with and without thiamine. When studied histochemically, soleus muscles from thiamine-deficient rats showed (1) small, angular fibers that had high NADH dehydrogenase activities; (2) a loss of 43% of type II (FOG) fibers; (3) decreased intensity of the reaction for betaOHB dehydrogenase; and (4) fibers with subsarcolemmal collections resembling "ragged-red" muscle. Electron microscopy revealed degeneration of some small myelin sheaths of distal and intramuscular nerves; atrophic, degenerating, hypoosmophilic muscle fibers in soleus and vastus medialis; and scattered muscle fibers with abnormal collections of deranged mitochondria accompanied by lipid droplets. These abnormalities, not found in control muscles, indicate that both motor neuropathy and mild mitochondrial changes, such as are seen in the "ragged-red" diseases, are induced by pure thiamine deficiency.


Recording site in organ of Corti determined by electrode marking technique. Microelectrodes containing 9% Niagara Sky Blue dye were used to record and electrophoretically mark cells in the guinea pig organ of Corti. The tissue was fixed in acidified glutaraldehyde and the marked cells were located using the surface preparation technique.


The infant development distress (IDD) syndrome: a predictor of impaired development? A group of eight infants with a disorder termed the IDD syndrome and another group of eight matched healthy infants taken into study at the same time have been followed-up for 6.75 years. The follow-up technique and some findings are described. Three IDD syndrome children are still severely disturbed. Three others show recovery from severe developmental disturbance. But one child who was grossly abused after being taken into the study has been well since the third follow-up. The comparison children have not experienced comparable problems. A qualified affirmative to the question of predictive power is given and discussed.


Esterase polymorphism and sensitivity to Dursban organophosphorus insecticide in Culex pipiens pipiens populations. Esterase polymorphism and Dursban (O,O-dimethyl-2-pyridylphosphorothioate) sensitivity have been investigated in 12 natural populations and three laboratory strains of Culex pipiens pipiens. This mosquito has two esterase loci, Est-1 and Est-2, which were shown to code esterases of the B group (aliesterases) but not cholinesterases. No correlation between Est-1 polymorphism and Dursban sensitivity was found, but the increase of the Est-2(0.64) allele in the populations less sensitive to Dursban was highly significant (r = -0.9850 for 6 df).


2-Amino-2-deoxygalacturonic acid in lipopolysaccharides from Pseudomonas aerugimosa. 2-Amino-2-deoxygalacturonic acid was identified as a component of the lipopolysaccharide from Pseudomonas aeruginosa N.C.T.C. 8505. The compound probably occurs in the region of polysaccharide responsible for O-antigenic specificity.


[A modified method for the determination of locations of chronically implanted electrodes in rabbit brains (author's transl)]. A fast and simple method is described for the localisation of tips of chronically implanted electrodes in the brain of rabbits which may be obtained--under standard conditions--by an intra-aortal injection of a 2% potassium-hexacyanoferrate-III solution. This method also facilitates histological preparations with a thickness of 5 mu only and thus permits a thorough morpho-pathological examination not only in the area of the electrode tips but in adjacent zones as well. The use of a buffered formalin solution also permits certain histochemical examinations to be made, for instance the determination of nucleic acids.


Inhibition of the effects of alfathesin and other steroid anesthetics by catatoxic steroids in rats. In rats, the sleeping time induced by overdosage with eight steroid anesthetics--alfathesin, 3-(3-oxo-17beta-hydroxy-19-nor-4-androsten-17alpha-yl)-propionic acid-lactone (SC-8109), 21-hydroxy=5alpha-pregnane-3,20-dione (P-234), 4-pregnene-3,11,20-trione (Bio.66), 17-hydroxy-3-oxo-4-androstene-17alpha-propionic acid-gamma-lactone(SC-5233),3alpha-hydroxy-5beta-pregnane-11,20-dione, 5beta-pregnane-3,11,20-trione (U-1373), and hydroxydione--was abolished or considerably reduced by a variety of catatoxic compounds, particularly 3beta-hydroxy-20-oxo-5-pregnene-16alpha-carbonitrile (PCN), 9alpha-fluoro-11beta,17-dihydroxy-3-oxo-4-androstene-17alpha-propionic acid potassium salt (CS-1), prednisolone, ethylestrenol and spironolactone. Phenobarbital and diphenylhydantoin, two non-steroidal stimulators of hepatic microsomal drug metabolism, were also highly effective. In contrast, triamcinolone, estradiol,progesterone, desoxycorticosterone and hydroxydione, which exert little or no catatoxic activity, failed to significantly diminish anesthesia or sedation.


Percutaneous absorption of 14C-9alpha-fluoro-11beta,21-dihydroxy-3,20-dioxopregna-1,4-dieno (17alpha,16alpha-d)-2'-methyloxazolidine-21-acetate (L 6400) in rats, pigs and humans. Comparison with 3H-triamcinolone acetonide. 1. The percutaneous absorption of a novel oxazolino corticosteroid, 14C-9alpha-fluoro-11beta,21-dihydroxy-3,20-dioxopregna-1,4-dieno[17alpha,16alpha-d]-2'-methyloxazolidine-21-acetate (L 6400) and 3H-triamcinolone acetonide (TA) was compared in rats, pigs and humans after application of a cream formulation to the skin and occlusion. 2. Within 96 h, up to 4.0% of the applied dose of L 6400 (permeability constant Kp 0.28) and up to 8.7% of TA (Kp 0.42) were percutaneously absorbed in rats. 3. Microhistoautoradiography showed that most of the applied radioactivity remained on the skin of rats. Radioactivity in the skin was mainly associated with the epidermis. 4. Within 120 h, up to 1.5% of the applied dose of L 6400 (Kp 0.16) and up to 0.5% of TA (Kp 0.07) were percutaneously absorbed in pigs. Plasma concentrations of radioactivity were maximal 48 h after application of 3H-TA whereas they were undetected after application of 14C-L 6400, because of the relatively lower specific activity of this 14C-steroid. 5. Both corticosteroids were allowed to remain in contact with human skin for 5 h and during that time, up to 1.1% of the applied dose of L 6400 (Kp 0.82) and up to 0.9% of TA (Kp 0.94) were absorbed. Results in four human subjects varied twenty-eightfold for L 6400 and fivefold for TA. Plasma concentrations of radioactivity were undetected after application of L 6400. In one subject treated with 3H-TA they were maximal after 2 h but were undetected in another subject and were found only in early samples from two others. 6. The results suggested that both corticosteroids were poorly absorbed through normal skin in rats, pigs and humans.


Electron microscopic demonstration of immunoglobulin deposition in rheumatoid cartilage. Horseradish peroxidase (HRPO) conjugated with goat antihuman IgG, goat antihuman IgM, and aggregated human IgG has been used as a enzymatic marker to stain IgG, IgM, and rheumatoid factor in rheumatoid cartilage. When Hrpo-anti IgG and HRPO-anti IgM were used, immunoglobulin deposits were not observed in nonrheumatoid cartilage. However 7 of 8 rheumatoid cartilage specimens stained with HRPO-anti IgG showed electron-dense deposits. Three rheumatoid specimens stained with HRPO-anti IgM showed similar findings. Both of 2 rheumatoid specimens also stained positively with HRPO conjugated with aggregated IgG, a finding indicating that rheumatoid factor was present. The deposits were seen between the collagen fibers of the superficial layer of the cartilage to a maximal depth of 22 mu from the surface (average: 7 mu). The amorphous fibrinous material on the surface of the cartilage was also stained. The demonstration of IgG, IgM, and rheumatoid factor in the superficial zone of rheumatoid cartilage suggests that immune complexes are deposited in the cartilage in this disease.


Effect of complement and polymorphonuclear leukocyte depletion on experimental skin lesions resembling systemic lupus erythematosus. Immunopathologic cutaneous lesions resembling human systemic lupus erythematosus (SLE) can be induced in mice sensitized to ultraviolet (UV)-irradiated DNA following whole body irradiation with UV light. The lesions are characterized by the formation of immune complexes at different skin sites. The role played by cellular and humoral mediators in the pathogenesis of this experimental model was investigated. The results obtained suggest that inflammation that follows UV radiation is the major factor responsible for this pathology. Accordingly mice that were rendered neutrophil PMN) deficient did not manifest skin lesions, and depletion of C3 complement component left them unchanged. In addition time course studies showed that PMN depletion did not prevent a delayed skin involvement. Thus multiple factors seem to mediate the onset of the immunopathologic changes previously described.


2-phenylethanol and some of its amphiphilic derivatives as inhibitors of platelet aggregation. Structure-activity relationship. The relationship between chemical structure and inhibitory activity of some simple water soluble phenylalkyl derivatives has been investigated. The inhibitory effect of the following analogous and homologous derivatives increases in the sequence: phenylacetic acid less than 2-phenylethanol less than 2-phenylethylamine = 4-phenylbutyric acid less than 3-phenylpropanol less than 4-phenylbutylamine, i.e., the inhibitory effect is potentiated. 1. by elongation of the alkyl chain, which is accompanied by an increase of the lipophilic character, and 2. when the polar portion is represented by the positively charged amino-group. The effect of these amphiphilic compounds on platelet aggregation appears to be mediated through polar and apolar interactions with the outer layers of the plasma membrane. The fact that the amino derivatives are the most active inhibitors suggests that negatively charged groups may be involved in the polar binding of these inhibitors. Experiments with neuraminidase-treated platelets revealed that the enzymatically removable sialic acid residues are not those negatively charged binding sites. Phentolamine and propranolol which block adrenergic alpha- and beta-receptors, respectively, do not influence the inhibitory effect of these agents.


[Characteristics of the phlogistic action of melittin, its C-terminal fragment and some basic peptides in the rat paw (author's transl)]. Injection into the rat paw of either melittin, its C-terminal dekapeptide, tetralysine or triarginine causes an inflammatory response with melittin having the highest and the last mentioned peptides the lowest activity. The inflammatory action of melittin and the dekapeptide has the following characteristics: 1. it is mainly due to the liberation of histamine, since on both compounds the anti-histaminic diphenhydramine has a pronounced inhibitory effect; 2. di-sodium-cromoglycate inhibits only the inflammatory action of the dekapeptide but has no effect on the activity of melittin. Melittin and its fragment thus liberate histamine via different routes.


Immunologic cell features in lymphocytic leukemias: a review. Since four years, it has been possible to classify most cases of Chronic Lymphocytic Leukemia (CLL) as given to the expansion of abnormal B-lymphocytes. On the contrary, the T-cell class is apparently normal and the T cell extent in CLL-peripheral blood can be even greater than normal when taken as absolute value. The clonal nature theory of B-lymphocytes in CLL is substantiated by the fact that, in general, in every patient only one Ig light chain determinant is present. Again, when serum Ig monoclonality is present in CLL, it appears idiotypically identical to the Ig shown by the lymphocytes of the same patient. Among the most important B-lymphocyte abnormalities in CLL, there are: (a) fluorescence of surface Ig is usually less intense than that in normal subjects, and fluorescence intensity may vary not only from patient to patient, but also from cell to cell in the same patient; (b) the Fc-receptor can be lacking; (c) the C3b-receptor is not always present, or it is from 2 to 20 folds less frequent than the C3d-receptor, whereas normal human lymphocytes do not show any outstanding differences between the number of EAC rosette-forming cells either when tested with mouse complement (C3d-receptor) or with human complement (C3b-receptor); (d) the traffic capacity of peripheral-blood B-lymphocytes in CLL is quite defective. All the above-mentioned data, taken as a whole, suggest that CLL is in general given by the expansion of an abnormal clone of cells of B origin, arrested in their maturative development, non-responsive to the mitogen stimulation, accumulating in the peripheral-blood for a traffic deficiency.


[The complement fixation test with BCG in the diagnosis of leishmaniasis]. The utility of complement-fixation test with an antigen from commercially lyophilized BCG was investigated in the diagnosis of leishmaniasis. The following results were obtained: Visceral leishmaniasis: 4 positivity on 5 cases; Oriental sore: 3 positivity on 6 cases; Canine leishmaniasis: 6 positivity, at high level, and 2 negativity; Other diseases: 5 positivity, on 17 sera of tbc; No positivity in sera from patients with various diseases, in control and in normal dogs. The results are compared with those quoted in medical literature and discussed. The meaning of the reaction is also briefly discussed, and the hypothesis of the presence of partigens between BCG and leishmania, or presence of autoantibodies is prospected.


[Induction of prophage gamma by means of bleomycin treatment]. The kinetics of induction of lambda prophage with bleomycin A2-CP3, mitomycin C and a combination of these two antibiotics have been studied. Bleomycin has been found to cause a rapid induction of prophage in exponentially growing cultures of E. coli K12 (lambda), and the maximal values of including activity are obtained after treatment for 16 minutes with a concentration of 10 mug/ml. The per cent of induction, calculated from the total number of surviving cells, including the phage producing cells, better expresses the dynamics of the phenomenon. The number of inducing events in the population of exposed cells is proportional to the length of bleomycin treatment. These values are greater than those observed after treatment with mitomycin, at the same concentration. Inducing activity is enhanced by combination of the two antibiotics and this effect is not additive, but synergic. After the 16 minutes of treatment with bleomycin the burst sizes are lower than those which are obtained with mitomycin or with the combination of the two antibiotics. The results are interpreted as a consequence and a confirmation of the different mechanisms of action that distinguishes bleomycin from mitomycin.


Natural antibodies against colicines in human and animal sera. The presence of "natural antibodies" against colicines was detected in the sera of 13 (29.5%) hospitalized recruits aged 20-29 years, 4 (25%) hospitalized males aged 30-49 years, 8 (40%) outpatient elderly subjects aged 50-79 years and in the sera of 15 (27.2%) animals. It is discussed the possible role of colicines as antigenic determinants stimulating the production of "natural antibodies", because of the prolonged and repeated contact of colicinogenic strains with the host animals.


Effect of thiouracil upon canine arterial elastic tissue. Unlike some other mammalian species, the dog is relatively resistant to the development of elevated levels of serum cholesterol after prolonged cholesterol feeding. This may be overcome by suppressing thyroid activity with thiouracil. Information regarding possible activity of thiouracil itself upon the arterial tissues is almost nonexistent. The present investigation was undertaken to test whether this drug has any such action, especially upon the arterial elastic tissues. Destructive changes were observed in arterial elastic tissues in dogs given thiouracil for three and six months. The changes consisted of accentuation of the elastic fibrillar components, formation and subsequent coalescence of clefts, and fragmentation and ultimate "dissolution" of the elastic elements. The results suggest that thiouracil may exert a damaging effect upon the arterial elastic fibers; thus, it is possible that one of the mechanisms by which thiouracil and cholesterol administration induces experimental atherosclerosis in the dog is by elastic tissue destruction, possibly promoting the subsequent lipid accumulation in the arterial wall.


Spread of hepatitis-B infections. A seroepidemiologic study. A solid-phase radioimmunoassay (R.I.A.), allowing simultaneous detection and quantitation either of HB, Ag or of anti-HBs, was applied to check the spread of hepatitis-B virus (H.B.V.) infections in various groups of population. R.I.A. procedures were also used to validate positive reactions and to determine the subtype specificity of sera. The overall prevalence of neutralizing anti-HBs in tested sera was almost 4-fold higher, although generally at rather low titres, than prevalence of HBs Ag. Frequency of HBs Ag and anti-HBs was rather high among healthy individuals from the normal Italian population and was extremely high among individuals from East Africa (Tanzania), as well as among healthy or sub-normal institutionalized subjects, among dialyzed renal patients and unselected blood-donors from 5 Italian towns. 80.2% of HBs Ag and anti-HBs detected in Italy were ay subtype, while most of positive sera from Tanzania (72.3%) were ad, which extends the map of H.B.V. subtypes in Africa. All tested ad antigens, either from Italy and from Tanzania, carried the w secondary determinant. Low-titre neutralizing anti-HBs, predominantly bearing a and d specificity, were also found in almost all examined preparations of standard immunoglobulins, which were imported from U.S.A.


Increase in serum alpha foetoprotein level in hepatic regeneration of the rat. Effects of age and of magnitude of regenerative activity. The effects of variable degrees of liver regeneration induced by two-thirds partial hepatectomy or intoxication with different doses of carbon tetrachloride (CCl4) on increased production of alpha foetoprotein (AFP) have been studied in very young (5-6 weeks) and older (15 weeks and more) rats by counter-immunoelectrophoresis (sensitivity down to 250 ng/ml). In the young animal adequate regeneration following two-thirds hepatectomy as well as 100 myl CCl4 successfully induced large increases in serum AFP levels. Smaller doses as well as a large fatal dose of the toxin, all of which resulted in inadequate regenerative activity, failed to excite AFP synthesis and secretion up to detectable levels. The adult animal did not show detectable AFP with any of these procedures. However, necrosis induced in the regenerating adult liver 24 h following partial hepatectomy did result in the detection of small amounts of protein in the serum. It is concluded that in hepatic regeneration of this species, the age of the animal as well as the magnitude of regenerating activity in the liver are critical in the occurrence of high levels of serum AFP.


Tape/slide programmes for independent study. Methods of presentation to students. Programmed audio tape/slide presentations have been presented to students by a variety of methods, (a) individual or pairs of students using automatic equipment in study booths; (b) students using simple portable equipment in or out of the learning centre; (c) groups of students in which discussion was encouraged; and (d) groups of students in which the student answered the questions individually without discussion. Each method has its advantages and disadvantages. With the exception of the groups without discussion, all were popular and chosen by a significant number of students as the method of choice. Automatic equipment in study booths was the most popular single method though the use of simple portable equipment had particular advantages. The majority of students prefer to have access to more than one method of presentation and attempts should be made to provide the learner with a variety of opportunities. The use of individual learning materials by groups of students should be further explored.


Measuring blood pressure. Training film to illustrate problems of intra- and inter-observer variation. Some of the errors inherent in the measurement of blood pressure and how they can be overcome have been described. A film has been made consisting of 16 blood pressure readings. This can be used in an attempt to illustrate to a class of students the problems of intra- and inter-observer variation. It is also suggested that the readings act as an easily collected source of data which can be used by students outside the class for further analysis.


Tape/slide course in ophthalmology for undergraduates. A tape/slide course in ophthalmology was used for fifth year medical students at the University of Edinburgh. A statistical comparison between the tape/slide and lecture method of teaching showed no significant difference in examination results: means mark (P greater than 0-50), spread of mark (P greater than 0-05), or in question consistency (P greater than 0-40). The tape/slide method is initially time consuming to produce but has clear advantages in the long term, being of a consistently high standard, easy to present and repeat, useful in revision, for student electives and in lecturers' absence.


Reflex disturbances of cardiac rhythm during ophthalmic surgery. Cardiac rhythm was monitored in 219 patients who had ocular operations under general anaesthesia and in 15 patients who had cataract surgery under local analgesia. Of these, 140 were children having squint surgery. The high incidence of the oculo-cardiac reflex and the adequate protective effect of atropine 0-01 mg/kg given intravenously was confirmed. Isolated extrasystoles were common, particularly in older patients, and sustained ectopic cardiac arrhythmias were observed in 4-6 per cent of patients of all ages. The significance of these arrhythmias in relation to surgical stimulation and the anaesthetic drugs used is discussed.


The immunological properties of pyruvate kinase. II. The relationship of the human erythrocyte isozyme to the human liver isozymes. We have examined the hypothesis that the human erythrocyte isozyme of pyruvate kinase (EC 2.7.1.40) is a hybrid of the two isozymes present in liver. Rabbit antiserum against purified human erythrocyte pyruvate kinase inactivates the erythrocyte isozyme and the major liver isozyme from human tissue but does not inactivate the minor liver isozyme. The electrophoretic mobilities of the erythrocyte and major liver isozymes are altered by anti-erythrocyte enzyme antibody while the mobility of the minor liver isozyme is unaffected. Gel diffusion analysis indicates cross-reactivity between the erythrocyte and major liver isozyme but no cross-reactivity with the minor liver isozyme. The hybrid hypothesis would predict cross-reactivity including changes in activity and mobility of all isozymes and we conclude, therefore that the hypothesis is incorrect.


Calf lens messenger ribonucleoprotein complexes. Characterization and comparison of template activity with corresponding mRNAs. Lens messenger ribonucleoprotein complexes have been isolated from calf lens polysomes by sucrose gradient centrifugation after puromycin-induced dissociation. A 10 S mRNA was released from a 13 S messenger ribonucleoprotein complex and a 14 S mRNA from a 19 S messenger ribonucleoprotein complex. Two major protein components with molecular weights of approx. 64 000 and 40 000 were isolated from each of the messenger ribonucleoprotein complexes after RNAase digestion. Buoyant density determinations suggest that the messenger ribonucleoprotein complexes contain approximately one mol of each major protein species per mol mRNA. In contrast to lens mRNA, lens messenger ribonucleoproteins are poor templates for transcription with avian myeloblastosis virus reverse transcriptase. Similar results were also obtained with globin messenger ribonucleoprotein containing either two major protein species (or deficient in the lower molecular weight protein species). Polynucleotide phosphorylase eliminates the reverse transcription template activity of the lens mRNA. This effect is blocked in the messenger ribonucleoprotein. Such observations suggest that at least one of the protein components associated with lens messenger ribonucleoprotein may be located in the 3'-terminal region. Only a small variation in translation activity was observed between the messenger ribonucleoproteins and their respective mRNAs.


What moves chromosomes, microtubules or microfilaments? An investigation of the spindle apparatus of crane-fly (Nephrotoma suturalis) spermatocytes has been undertaken using methods that permit combined light and electron microscopy of selected cells. At the ultrastructural level, spindles contain microtubules in a granular matrix. Microtubules have been classified as kinetochore microtubules (which connect to kinetochores of chromosomes) and non-kinetochore microtubules (not attached to kinetochores). Kinetochore microtubules are distributed in densely packed bundles, which are the birefringent chromosomal fibers seen in living cells. Actin filaments were not observed in spindles of unglycerinated cells or in cells fixed in glutaraldehyde containing tannic acid, which negatively stains F-actin in situ and thus can be used to aid the localization of actin filaments in non-muscle cells. The absence of actin filaments in the spindle coupled with their presence in the "contractile ring" of spermatocytes fixed during cytokinesis is evidence against the hypothesis that chromosome movements are microfilament-based. The results are compatible with the hypothesis that microtubules are involved in the mechanism of chromosome transport. The details of that mechanism remain to be clarified.


[Effect of synthetic polynucleotides and RNA on poly(C)-dependent poly(G) polymerase activity of Q beta replicase]. The effect of synthetic polynucleotides and phage RNA on poly(C)-dependent synthesis of poly (G) by Qbeta replicase is studied. It is shown that single stranded poly(U) and poly (dT) are strong inhibitors whereas structured polynucleotides poly(A), MS2 RNA as well as double stranded complexes poly(A)-poly(U) and poly(A)-poly(dT) do not affect the synthesis of poly(G). It is suggested that contact region of template with enzyme has single stranded unhelical structure and affinity of polynucleotides to Qbeta replicase is determined by degree of their secondary structure.


Evidence for significant hematopoiesis in the human thymus. Earlier studies on fetal thymus suggested that certain of the large pyroninophilic cells found there might have a hemopoietic role, and it was decided to determine the nature of these cells using histochemical and immunohistochemical methods. Thymic tissue from aborted fetuses, stillbirths, and neonatal deaths was examined histochemically using methods for the detection of chloroacetate esterase, peroxidase, and pseudoperoxidase, and by staining techniques for mast cells and eosinophils. Tissue was also examined using the indirect immunoperoxidase method for the presence of hemoglobin A (HbA) and F (HbF), for lysozyme (muramidase) and immunoglobins alpha, mu, gamma, kappa, lambda. Positive staining to some degree was seen in cells in the connective tissue stroma using all methods, and the cells stained corresponded to one or another of the types of pyroninophilic cells present. The finding of large cells with positive chloroacetate esterase and antilysozyme indicates the presence of granulopoiesis. Similarly, the presence of large nucleated cells with pseudoperoxidase and anti-hemoglobin (A and F) staining indicates the presence of erythropoiesis. Plasma cells were present in small numbers.


Polychlorinated biphenyls (PCB's) in the rivers Avon and Frome. On June 19, 1973 very high levels of Aroclor 1254 were determined in the River Avon and Frome. The source might have been cleaning of systems in factories which use them. The levels decreased with time. In fish from the River Avon three types of Aroclors were determined.


Serum protein analysis following implantation of methyl methacrylate in rats. Thirty-six Sprague-Dawley white rats were implanted with methyl methacrylate and bone wax in a 1:1 ratio. At periods of 1, 3, 5, 7, 10 and 14 days-post-implantation arterial blood samples were drawn and the serum protein separated electrophoretically. A significant depression in the alpha 1 and alpha 2 fractions of the methacrylate animals was demonstrated as was a significant increase in the beta fraction of the 3 day group. There was a sustained depression of the alpha 1 fraction in the methacrylate animals in a 3 day and a 5 day time period. It is not felt, however, that this is indicative of a mechanism for the diverse phenomena that have been described secondary to implantation of methyl methacrylate.


Differentiation of squamous carcinoma of the larynx as a determinant of prognosis. The changes seen by light and electron microscopy in the differentiation of squamous carcinoma are described. The grading of the differentiation of squamous carcinomas by overall impression and by quantitative methods are discussed. Reasons are given for retaining the former in the assessment of prognosis. Other differentiating phenomena, particularly retention of basement membrane in the formation of rounded rather than jagged growing edges, are discussed. The differentiating effect of radiation on squamous carcinomas is discussed as a means of assessing the efficacy of the radiation. A similar differentiating effect of bleomycin, a cytotoxic drug, on human squamous carcinoma is described. Using electron microscopical criteria belomycin also increases differentiation of a transplanted, experimental, poorly differentiated squamous carcinoma and human laryngeal carcinoma cells in tissue culture. The significance and possible practical utilization of this phenomenon are discussed.


"B" cell origin of malignant cells in a case of American Burkitt's lymphoma. Characterization of cells from a pleural effusion. Ninety-eight percent of the cells isolated from a malignant pleural effusion in a case of American Burkitt's lymphoma showed membrane fluorescence to antihuman IgM antisera, while only 2% of the cells formed spontaneous rosettes with sheep red blood cells. It is concluded that the Burkitt cells in the malignant effusion are derived from bursal equivalent lymphocytes. Epstein-Barr (E-B) viral titers to both viral capsule antigen and early antigen were not elevated. The significance of these findings is discussed.