Alcoholic hyalin antigen (AHAg) and antibody (AHAb) in alcoholic hepatitis. Complement fixation (CF) and immune adherence (IA) hemagglutination tests demonstrate antigen (Ag) and antibody (Ab) to alcoholic hyalin (AH) in serum of patients with alcoholic hepatitis. Immunoglobulins from postmortem liver of patients dying of alcoholic hepatitis show antibody activity against AH. Isolated purified AH was used to produce AHAb in rabbits. Rabbit antiserum was added to heat-inactivated human serum to detect AHAg; purified AH was added to serum and tissue elutes to detect AHAb. AHAg was present in serum of each of 15 patients with early phase alcoholic hepatitis in CF titers of 8 to 64 and IA titers of 16 to 2048. AHAg became negative within 3 to 5 weeks with abstinence from alcohol and was followed by transient appearance of AHAb. AHAb was present in 11 patients with advanced phases of alcoholic hepatitis in CF titers of 8 to 640 and IA titers of 16 to 4096. Four patients in this group exhibited concomitant AHAb and AHAg. Investigations of liver tissue elute reveal that patients with advanced alcoholic hepatitis or active alcoholic cirrhosis have AHAg-reactive immune complexes containing IgG and IgA immunoglobulins.


Chemical nature of alcoholic hyalin.

Chemical nature of alcoholic hyalin. Alcoholic hyalin was prepared from postmortem livers of patients with alcoholic liver disease. Alcoholic hyalin was purified by discontinuous sucrose density gradient ultracentrifugation. The purified protein preparation contained about 12% reducing sugar, the remainder being protein. Gel filtration on Sephadex 4B column showed the presence of three peaks. Each peak material thus obtained contained acetylglucosamine, hexosamine, and deoxysugar (calculated as rhamnose). That these protein components followed by scanning the gels at 280 nm, and at 520 nm after staining the gels with periodic acid-Schiff base reagent for the presence of carbohydrate.


[Identification of monkey chromosomes by differential staining with Romanovsky--Giemza stain. III. Cercopithecus aethiops]. The distribution of the G-bands in chromosomes of bone marrow cells of Cercopithecus aethiops was studied by means of differentail staining with the Romanovsky - Gimsa dye. All the homologous chromosome pairs were identified and morphometric parameters were detected. The comparison of C. aethiops karyotypes with those of Macacca mulatta has shown that they are different in numbers and in the character of banding pattern of the most chromosomes. Both species revealed 12 pairs of chromosomes similar in their morphology and parameters.


Time-conditioned coordination of dehydrogenase activity in blood cells in infantile acute leukemia. The correlation of enzymatic activities studied at different periods of time (lasting from several hours to several days) has been revealed in 39 children at different stages of acute leukemia and in 8 children suffering from other hematological diseases (Hodgkin's disease, Morbus Werlhof, reactive reticulosis). The correlation of activity of enzymes constituting different systems and arranged in different parts of cell organelles enables us to explain this phenomenon by a regulation on a higher level, viz. cytophysiological processes as a whole. The coordination of enzymes possible depends on the synchronism of enzymatic activity caused by the physiological process developing in the cell.


[The effect of extracorporeal blood radiotherapy on the peripheral blood T and B-lymphocytes in chronic lymphadenitis]. B-lymphocytes and T-lymphocytes were determined before and after therapy in 12 patients with chronic lymphatic leukemia (CLL) treated with extracorporal blood radiation (ECIB). There was a significant decrease of B-lymphocytes (p less than 0.001) from 85.3 +/- 6.55% to 71.55 +/- 10.60% by ECIB, whereas no significant changes could be found in T-lymphocytes. 12 untreated CLL patients, whose B-lymphocytes amounted 83.25 +/- 6.79% with 7.5 +/- 3.42% of T-lymphocytes, were examined as group of comparison. From these findings ECIB is concluded to cause a decrease of accumulated B-cells.


[Morphological, cytochemical autoradiographic and quantitative studies of the cells in lymphocyte cultures with added tuberculin]. Lymphocyte culture experiments with purified tuberculin showed a transformation of small lymphocytes into medium sized, large and very large basophilic cells. During cell growth RNA synthesis and acid phosphatase activity increased. Some of the very large basophilic cells synthesized DNA. During the first and second day of culture experiment also medium sized and large weakly basophilic cells increased, which have a pale plasma and azurophilic granules. These cells did not synthesize DNA and showed only a low incorporation of 3H-uridine. The results of our experiments indicate a transformation of medium sized and large basophilic cells not synthesizing DNA into weakly basophilic (pale) cells showing azurophilic granules. In contrast to the lymphocyte culture experiments with tuberculin lymphocyte cultures without tuberculin showed no significant increase of medium sized, large and very large basophilic cells and of weakly basophilic cells.


[A case of IgD plasmacytoma with unusual neurological symptoms]. A disease in an IgD (lambda) plasmocytoma is described, where after therapy with Alkeran and prednisone a disappearance of all clinical and laboratory findings indicating an activity could be observed. However, there was a progressive development of a picture of encephalomyelitic and polyradicular neuritic syndrome in the female patient with all signs of a recidivous temporary intracranial hypertension which could not be cured. The aetiology of these processes could only be found by the autopsy which revealed an isolated massive infiltration of the meninges and the sheaths of the spinal cord nerves with atypical plasma cells. No signs of the tumour could be identified in other organs. Such localization of the disease, as it is described here, is the first observation of its king. Problems of clinical diagnosis and treatment are discussed.


[Cytoplasmic vacuoles and kinetics of bone marrow cells in alcoholism, liver cirrhosis and anemia]. Bone marrow smears show vacuoles in the cytoplasm of haematopoietic cells due to alcoholism, liver cirrhosis and anemias different origin, for example in acute leukemias, in proportion to the decreased content of haemoglobin. The cell doubling rat in vivo is not obviously impaired by the vacuolisation of the cytoplasm exactly as is the case with proliferation in vitro, although the vacuolation increases in vitro. Phase contrast observation makes visible the vacuoles without membranes and content as they appear transitorily in pinocytosis, and not as secondary lysosomes. The common cause for the formation of the vacuoles without membranes in the cytoplasm in all these pathological states could be intracellular hypoxia and acidosis.


[The effect of storage and short term cultivation the proliferation of mouse bone marrow stem cells]. Suspension of stored and short-term cultured murine bone marrow cells was i. v. administered to lethally irradiated mice. We demonstrated that the cells still after 72 h of storing and 24 h of culture in liquid medium were capable of proliferation and on day 9 following transplantation could produce colony forming units in the spleens of lethally irradiated mice. We established the relative participation of individual types of cells in the recovery of haemopoiesis in the spleens.


Conductance of the cytoplasm of erythrocytes as a criterion for evaluating the quality of blood preservation. It has been established that enzymic systems become exhausted during the preservation of blood, which causes disturbances in the transport of ions across the cell membrane. These changes occurring in the cell may be recorded by measuring the electrical conductance of cytoplasm. The conductance has been determined for three selected preserving liquids in relation to the blood storage time. Results of this study showed that the dynamics of changes in the electrical conductance of the cytoplasm is characteristic for a given type of the preserving liquid. Hence, the determination of changes in the electrical conductance of the cytoplasm may provide a new important criterion for evaluating various methods of blood preservation.


[The effect of the AcD-AG stabilizer on the survival time and surface properties of fluid preserved platelet concentrates compared to concentrates from fresh blood]. In the present investigations the storage effect the AcD-AG stabilizer on thrombocytes is examined. The thrombocytokinetic parameters of 9 fresh blood concentrates and 15 concentrates of AcD-AG plasma containing platelets were determined. Storage time amounted to three days. The results show that storage with AcD-AG is only possible to a limited degree. On an average the survival time of the platelets was reduced to 2.7 +/- 1.1 days compared with 9.0 +/- 1.0 days of fresh blood concentrates. The recovery of the stored platelets amounted to 25.3 +/- 16.1%, that of the fresh blood concentrates to 63.3 +/- 23.6%. The spleen-heart quotients and those of the liver-heart or the surplus impulses over the spleen and liver respectively indicate that there is a predominant destruction in the spleen for those thrombocytes stored for three days. The liver is scarcely involved in this sequestration process. With 36.1% platelet yield was very low in concentrates gained from AcD-AG plasma containing platelets and having been stored for 3 days. In cases of emergency a clinical application of concentrates prepared in this way should not be given up. If being used, the greater requirement has to be taken into account. If the substitution therapy is continued, however, fresh blood concentrates have to be used as soon as possible.


[The diagnosis of vitamin B 12 deficiency]. In 5 normal persons, 1 patient with pernicious anemia, 5 patients with renal insufficiency in the state of compensation, and in one patient with chronic hemodialysis Schilling tests, activity of 57Co and vitamin B12 serum levels were measured. It was obvicous that activity of 57Co in serum 8 hours after oral application of 0.5 muCi 57Co-vitamin B12 showed reliable results to clear the problem of vitamin B12 malresorption. This method is easier than the Schilling test because the collection of 24-hour-urine is not necessary and an insufficiency of renal function does not influence the result.


Lysis of red blood cells in some haemolytic anaemias--the lytic effect of agkistrodon piscivorus venom in vitro. The lysis of red blood cells induced by the Agkistrodon piscivorus venom (APV) in vitro in the presence and/or in the absence of plasma was examined in autoimmune haemolytic anaemia (AIHA), hereditary spherocytosis (HS) and paroxysmal nocturnal haemoglobinuria (PNH). Virtually haemolysis did not differ from that of normals in AIHA, it was slightly increased in HS and significantly higher in PNH, however, only in the presence of autologous or normal homologous plasma. The mechanism of PNH blood cells lysis which is probably related to the activation of the third complement component is discussed.


[Quantitative studies of the ABO blood group system. Quantitative changes in agglutinability of antigens A, B and H with regard to the patient's age]. The results of a comparative quantitative investigation about the agglutinability of erythrocyte antigens A, B, and H to be found in fetus (fifth to ninth lunar month), newborns, adults, and old age people are represented. In investigating with anti-H sera it could be found that A-antigens in fetus undergo the same development to be observed in newborns; it amounts to 75% of the agglutinability present in adults. After delivery the agglubinability will essentially increase up to the fifth month; it will only reach the average values of adults, however, after the seventh year of age and will remain unchanged then until the end of life. The agglutinability of B-erythrocytes with anti-B will also change in the same way. In fetus and newborns the agglutinability of A1 erythrocytes with anti-A1 sera is markedly weaker than that determined by anti-A sera. After delivery it will rapidly increase, will be stronger afterwards than the agglutinability with anti-A sera and will have the values to be found in adults after the third year of age. After the 85 year of age, however, there is a tendency of weakening the agglutinability with anti-A1 serum. With growing age the agglutinability of O-erythrocytes with anti-H serum is changed in the same way. Furthermore, it could be detected that the interaction between genes A1 and B to be found in adults will find its expression in a weakening of the agglutinability of A1B-erythrocytes with anti-A1 and anti-B sera, the agglutinability with anti-A serum remaining constant. This behaviour is valid for the whole life of man. In all stages of life an interaction between weak and strong genes can be observed in A2B-erythrocytes.


[Studies on families with Osler's disease]. The authors examined 143 members of a family, where they found 37 (= 25.87%) of Osler patients. It was only in 30% of these patients that the symptoms occurred before the tenth year of age. As in one patient the symptoms did not appear until the age of 58 years, the possibility cannot be excluded that symptoms of the disease will become manifest even in other, sill younger members of the family in the course of time. Epistaxis was observed in 93% of the cases, nephrorrhagia in no case, hepatopathy and gastrorhagy were found only once in each case. The X-ray examination revealed arteriovenous pulmonary aneurysm in 5 cases. As a rule, oestrogen treatment led to good results. A case of death occurred during an influenza epidemic in a severe anaemic patient. Clinical main symptoms of Osler's disease were epistaxis and arteriovenous fistulae which could be roentgenologically identified in the lung. Teleangiectasia could be detected during the autopsy besides vessel anomalies on the surface even in the bronchi, oesophagus, trachea, stomach, kidneys, small intestine and particularly in the large intestines. Conditions of iron deficiency may very often occur in osler Patients; they require a substituting treatment.


Thrombotest mixing experiments in congenital coagulation disorders of the prothrombin complex and in coumarin treated patients. An additional evidence against the presence of an inhibitor in the latter. Thrombotest clotting times of mixtures of coumarin plasmas and normal plasma yielded a patterm similar to that observed in mixtures of plasma with congenital coagulation disorders and normal plasma. The presence of 10 or 20% of test plasma in the mixture failed to affect the clotting times which resulted in normal limits. The only exception to this rule was the hemophilia BM plasma. In this case even the presence of 10-20% of patient plasma in the mixture caused a prolongation of the clotting time. This indicates that no inhibitor is present in coumarin plasmas and in the plasma of congenital coagulation disorders of the prothrombin complex save for hemophilia BM plasma which does contain an inhibitor.


[Structural proteins of C-type RNA tumor viruses. Properties and significance for tumor immunity]. Five main structural polypeptides of Friend murine leukemia virus, a typical C-type RNA tumor virus, have been isolated. Their localization within the particle was attempted. They range from 10,000-71,000 in molecular weight and display three kinds of immunological determinants. The major envelope glycoprotein binds to cellular receptors and elicits the formation of antibodies which neutralize the virus and kill virus producing tumor cells in vitro. Natural as well as artificial immunity to this component protects mice against virus induced leukemia.


[The anaphylactoid reaction after colloid infusion]. All colloidal volume substitutes carry the risk of anaphylactoid reactions. The general incidence of 0.03% is low; however severe complications can occur. There are differnet pathomechanisms involved in the induction of anaphylactoid reactions after infusion of different colloids. Protein aggregates are of pathogenic importance in human serum albumin incompatibility. The addition of stabilizers to the plasma protein solutions can alter the immunogenicity of the protein molecule. Antibodies of the classes IgG and IgM seem to play a role in severe dextran incompatibility. There was a direct correlation of the intensity of the clinical symptoms with the anti-dextran-antibody titer.


Lymphoid cells as effectors of immunologic cytolysis. Two classes of cytotoxic lymphocytes are described. One is composed of cytotoxic T lymphocytes (CTL), which have antigen specificity derived from prior immunization. The lymphocytes of the other class--the K cells--derive their limited specificity from affinity for antibodies on the surface of the target cells. Together these two lymphocyte classes provide a broad-spectrum defense against unwanted foreign and mutant cells.


The vitamin D binding of the common and rare variants of the group-specific component (Gc). An autoradiographic study. The vitamin D3 binding properties of the common and rare Gc variants were examined. Vitamin D3 labeled with 14C was added to serum. Gc phenotypes were demonstrated autoradiographically following separation by immunofixation electrophoresis on agarose. This qualitative analysis did not reveal differences in vitamin D3 binding by the group-specific components of the common types Gc 1-1, Gc 2-1, and Gc 2-2. The double-band variants Gc Darmstadt, Gc Y/Ab, Gc Toulouse, Gc Norway, and Gc Caucasian were examined; the phenotypes Gc Ab-Ab, Gc Ab-1, Gc Ab-1, Gc Ab-2, Gc T-1, Gc T-2, Gc Norw-2, and Gc 1-Cau showed normal D3 binding. The double bands of Gc Darmstadt in the phenotype D-2 appeared somewhat weak. The single-band mutants Gc Wien, Gc Chippewa, Gc Opava, and Gc Z were analyzed; the phenotypes Gc W-1, Gc W-2, Gc Chip-1, Gc Chip-2, Gc 1-Op, Gc Op-2, Gc 1-Z, and Gc 2-Z showed normal D3 binding. A mutant in the Gc system with clearly defective vitamin D3 binding properties remains to be delineated.


Detection of Epstein-Barr virus antigens with enzyme-conjugated antibody. Monospecific conjugated (fluorescein isothiocynate and horseradish peroxidase) goat antisera, prepared against three human immunoglobulin classes, IM (mu), IgG (gamma) and IgA (alpha), were compared for their ability to detect human Ig classes possessing specificity for Epstein-Barr virus (EBV) viral capsid antigens (VCA) in a chronically infected human lymphoid cell line, P3J-HRIK. It was determined that the enzyme system was significantly more sensitive than immunofluorescence in detecting most of the immunoglobulins in sera from cancer patients. Some patients with nasopharyngeal carcinomas (NPC) had extremely high levels of EBV-specific IgA, whereas cancers other than NPC may have lower EBV-specific IgA titers.


Interspecies-, species- and type-specific T antigenic determinants of human papovaviruses (JC and BK) and of Simian virus 40. Immunofluorescence tests, absorption studies and quantitative analysis by a very sensitive 51Cr microcomplement fixation (CF) technique were used to define the degree of relatedness between the tumor (T) antigens induced by human papovaviruses, strain JC and BK, with simian virus 40(SV40) and mouse polyoma virus (PyV). Antisera against JCV, BKV, SV40 and PyV T were raised in tumor-bearing hamsters. The data obtained indicate that T antigens of JCV, BKV and SV40 possess various subspecificities which can be distinguished and looked upon as interspecies-, species- and type-specific antigenic determinants. It was found that JCV T and BKV T synthesized in transformed hamster cells share about the same amount (20%) of interspecies cross-reacting antigen with SV40 T from H-50 cell extracts (transformed hamster cells). Although hamster cells transformed by PyV showed definite PyV T reactivity, no cross-reactivity, at least with the sera used, was found with human papovavirus and SV40 T antigens. Furthermore, degree of heterogeneity was observed within the T antigen complex derived from different SV40-transformed cells.


Inhibiting effect of non-steroidal anti-inflammatory drugs on the chemotaxis of human leukocytes in vitro. Chemotaxis of human leukocytes was inhibited in vitro by four non-steroidal anti-inflammatory drugs, namely, lysine acetylsalicylate, phenylbutazone, indomethacin, and indoprofen. Dose-dependent effects were always found and significant regression was proved for all drugs except phenylbutazone.


Hydroxyethyl starch-induced macroamylasemia. Summarizing, the following results were obtained: 1. After infusion of 500 ml hydroxyethyl starch 6% in all patients an increase of serum amylase occurred, which in most cases reached values twice as high as the basal value. 2. This hyperamylasemia is caused by the formation of a high molecular HES-amylase complex, which cannot be easily eliminated. 3. In no case was hyperamylasemia associated with other clinical signs or symptoms.


Epidermal antigens.

Epidermal antigens. Autoantibodies in man may develop to antigens present in all of the major anatomical areas of the epidermis, namely the nuclei and cytoplasm of keratinocytes, the dermal-epidermal junction, the intercellular substance, and the stratum corneum. The antigens in all of these areas are polymorphic. Some are unique to man and to skin, others are present in other mammals and in other tissues. The biochemical nature of these antigens is mostly unknown. Several appear to be products of epidermal cell differentiation. Humoral immune reactions to some intercellular antigens is associated with pemphigus, to some basement zone antigens with bullous pemphigoid, and to some, cytoplasmic antigens with malignancies. The clinical significance of antibodies to the other epidermal antigens is still to be determined.


Cheyletiella dermatitis.

Cheyletiella dermatitis. A mild dermatitis probably due to Cheyletiella yasguri was observed in 2 persons whose pet dog was infested with this parasite. Cheyletiella mites differ morphologically from sarcoptic mites, which cause canine and human scabies. Treatment of man and dog with 1% gamma benzene hexachloride is usually successful in clearing this condition.


Comparison of methods for rapid immunofluroescent staining of group A streptococci and Neisseria gonorrhoeae. Immunofluorescent staining reactions for group A streptococci and Neisseria gonorrhoeae were equivalent to those of standard "slow" methods when slides were heated on a serological water bath at 50 C for periods one to four minutes. No false negative reactions occurred when this method was used. False negative reactions occasionally occurred when previously described rapid methods which employed a slide warmer were used.


Bacterial gram staining by conventional and strip methods. Conventional gram staining was compared with a method which uses dye impregnated paper strips for staining. The paper strip technique was judged accurate and reproducible and is recommended for use in the small laboratory and hospital ward.


N-banding and nucleolus organisers in Asellus aquaticus (Crust. Isop.). The N-banding technique was used to stain the nucleolus organiser of the karyotype of Asellus aquaticus (crust. Isop.). Observations were made on the morphological expression of nucleolus organisers as secondary constrictions and the presence of nucleoli in mitotic prophase. An attempt was made to correlate the various results and it seems likely that N-banding is not a reflection of NO activity.


Ultrastructural demonstration of cell coat on the cell surfaces of normal human oesophageal epithelium. The cell coat in human oesophageal biopsies was studied with Alcian Blue, Ruthenium Red, Safranin O, colloidal iron and the ferrocyanide--osmium tetroxide techniques. Alcianophilic material was found on the cell surface of the basal, prickle cell and functional layers, being most abundant on the superficial cells where it appeared as a continuous coat. In the deeper layers, it tended  to have a particulate distribution. Some membrane-coating granules were alcianophilic. Ruthenium Red had a particulate distribution over all cell surfaces. Intercellular debris was also stained. Safranin O produced no staining. Colloidal iron stained the cell coat in a particulate manner. The ferrocyanide--osmium technique showed a uniform filamentous cell coat. The oesophageal epithelial cell coats are, in part, acid mucosubstances which, on the surface cells, may have a protective function.


Some aspects of the value of Sudan Black B in lipid histochemistry. The lipid dye Sudan Black B, as generally used to demonstrate lipids in the interior of the cell nucleus, was studied with regard to its staining properties for isolated nuclei in relation with its chromatographic characteristics in solution, as well as with a model system consisting of lipid containing polyacrylamide films. Isolated nuclei are stained with Sudan Black B dissolved in ethylalcohol, only when the dye-solution is at least one month old. Extraction with chloroform-methanol (2:1) before staining resulted in a decrease of 35% in intensity. Treatment with proteolytic enzymes and DNA-se caused a complete disappearance of the staining capacity. The binding of Sudan Black B with phospholipids enclosed in the form of liposomes in modelfilms when stained with this dye in ethylene glycol obeys the law of Lamber-Beer, Proteins were however, also coloured by the dye. The chromatographic experiments showed that the dye is built up from two main and a number of secondary products. The secondary products which increase by aging of the dye-solution, change the spectrophotometric properties of the total dye and show a specific binding. The conclusion was reached that on the basis of a positive reaction with Sudan Black B no definite conclusions can be drawn about the presence of lipids in the interior of the cell nucleus.


Mucosubstances in the normal human oesophageal epithelium. A comparison of periodic acid-silver and phophotungstic acid techniques. Normal human oesophageal epithelium was investigated with the periodic-acid-silver methenamine technique and its variations to demonstrate neutral mucosubstances at the ultrastruct level. The results were compared with the acid phosphotungstic acid method. Neutral mucosubstances were shown in the cell coat and membrane coating granules by both techniques. The silver methods also demonstrated glycogen, the Golgi apparatus and dense bodies. The periodic acid-silver methenamine technique outlined positive material in the intercellular space of the prickle cell layer, but the other silver methods did not.


Immunological study of the regulation of cellular arylsulfatase synthesis in Klebsiella aerogenes. Regulation of cellular arylsulfatase synthesis in Klebsiella aerogenes was analyzed by immunological techniques. Antibody directed against the purified arylsulfatase from K. aerogenes W70 was obtained from rabbits and characterized by immunoelectrophoresis, double-diffusion, quantitative precipitation, and enzyme neutralization tests. Arylsulfatase was located in the periplasmic space when the wild-type strain was cultured with methionine or with inorganic sulfate plus tyramine, but not with inorganic sulfate without tyramine, as the sole sulfur source. Tyramine oxidase was retained in the membrane fraction prepared from cells grown in the presence of tyramine. Arylsulfatase protein was not synthesized in the presence of tyramine and inorganic sulfate by mutant K611, which is deficient in tyramine oxidase (tynA). We conclude that the expression of the arylsulfatase gene (atsA) is regulated by the expression of tynA and that inorganic sulfate serves as a corepressor. In addition, strains mutated in the atsA gene were analyzed by using antibody.


[3H] dihydrostreptomycin accumulation and binding to ribosomes in Rhizobium mutants with different levels of streptomycin resistance. Rhizobium trifolii B1, a symbiotic nitrogen fixer, is sensitive to streptomycin (10 microgram/ml) and spontaneously produces spheroplast-like forms during cultivation. Streptomycin-resistant mutants selected with high doses of antibiotic (1,000 microgram/ml) showed pleiotropic changes, including loss of spheroplast formation and infectivity to plants, whereas mutants selected with low doses of streptomycin (10 to 100 microgram/ml) retained properties of parent strain B1 (I. Zelazna-Kowalska, Acta Microbiol. Pol., in press). The present studies revealed that strain B1 and its mutant with a high level of streptomycin resistance, B1 strH, accumulated the antibiotic at similar rates. Mutant B1 strL, with a low level of streptomycin resistance (up to 100 microgram/ml), accumulated the antibiotic at a lower rate. Ribosomes isolated from strains B1 and B2 strL bound [3H]dihydrostreptomycin, whereas those from strain B1 strH did not. These observations indicate that, in R. trifolii B1, mutation to a high level of streptomycin resistance affects ribosomal structure, whereas low-level resistance involves a change in membrane permeability.


Characterization of the fatty acid-sensitive glucose 6-phosphate dehydrogenase from Pseudomonas cepacia. The adenosone 5'-triphosphate-insensitive glucose 6-phosphate dehydrogenase from Pseudomonas cepacia has been found to be strongly inhibited by long-chain fatty acids and their acyl coenzyme A esters, suggesting that an important role of this isoenzyme might be to provide reduced nicotinamide adenine dinucleotide phosphate for reductive steps in fatty acid synthesis. The enzyme, which has been redesignated the fatty acid-sensitive glucose 6-phosphate dehydrogenase, has been purified to homogeneity using affinity chromatography with nicotinamide adenine dinulceotide phosphate-substituted Sepharose as a key step in the purification. The purified preparations were used to study the immunological properties and subunit composition of the enzyme and its relationship to the adenosine 5'-triphosphate-sensitive glucose 6-phosphate dehydrogenase present in extracts of P. cepacia. Although both enzymes were found to be composed of similar size subunits of about 60,000 daltons, immunological studies failed to demonstrate any antigenic similarity between them. Studies of the sedimentation behavior of the fatty acid-sensitive enzyme in sucrose gradients indicated that its apparent molecular weight is increased in the presence of glucose 6-phosphate and suggest that it may exist in an aggregated state in vivo. Palmitoyl coenzyme A, which strongly inhibited the enzyme, failed to influence its sedimentation behavior.


Changes in enzyme activities of thymidine kinase and 5'-nucleotidase for dTMP during hormonal regeneration of seminal vesicles of mice. An increase of thymidine kinase [EC 2.7.1.21] activity and decrease of 5'-nucleotidase [EC 3.1.3.5] activity for dTMP were found during hormonal regeneration of the seminal vesicles by daily or single administration of testosterone propionate into mice castrated 2 weeks previously. Actinomycin D injected on day 0 of testosterone treatment completely inhibited both the increase of thymidine kinase and the decrease of 5'-nucleotidase. When injected on day 2, actinomycin D decreased thymidine kinase activity below the control level and 5'nucleotidase activity was not restored to the normal level. The activity of 5'-nucelotidase in a mixed sample, in which seminal vesicles of castrated mice and those of testosterone-treated mice were homogenized together, was intermediate between the activities determined separately. This indicates the absence of any inhibitor of 5'nucleotidase in the regenerating vesicles. Changes in total activity of 5'nucleotidase and total protein content in extracts during various treatments showed that the decrease in specific activity of 5'-nucleotidase in the first 2 days of testosterone treatment was not due to inhibition of enzyme activity but to dilution of the enzyme with other proteins which increased in content more rapidly than 5'-nucleotidase.


Estimation of internal pH in cells of blue-green algae in the dark and under illumination. Carbonylcyanide m-chlorophenylhydrazone (CCCP) or nigericin induced translocation of H+ In the dark across the cell membrane of blue-green algae Plectonema boryanum and Anacystis nidulans. The direction of the H+ flux depended on the pH of the suspending medium. At acidic pH, an influx of H+ and at alkaline pH an efflux of H+ were observed. It is suggested that the influx takes place at pH'S higher than the "internal" pH and the efflux at pH's lower than that. The internal pH was estimated to be 7.4+/-0.2 for Plectonema boryanum and 7.5+/-0.1 for Anacystis nidulans. Similar H+ changes due to CCP were observed under illumination, where the light induced efflux of H+ was limited by the counter-flux of cations. The internal pH of cells in the light, estimated from the pH-dependent reversion in the rate of the H+ change, was about 8.5.


Complexing Rauscher leukemia virus reverse transcriptase with human plasma ribonuclease from Hodgkin's disease patients. Human ribonucleases were purified from the sera of Hodgkin's disease patients by sequential column chromatography. The purified enzyme interacted with reverse transcriptase of Rauscher leukemia virus and formed an additive complex of Mr = 130,000. RNase and oligo(dG)-directed reverse transcriptase activities were diminished in the complex. The complex could be dissociated with the subsequent restoration of both activities in the presence of spermidine. The molecular weight of the complex suggest that the 2 RNase molecules bind to a single reverse transcriptase molecule.


Application of quantitative high-performance thin-layer chromatography in the antibiotic industry. For the in-process control of antibiotic fermentations and for routine assays of samples for scaling up and for pure products, quantitative high-performance thin-layer chromatography (HPTLC) can be used with advantage. Rapid chromatography on high-performance layers, combined with an automatic spraying device for exact derivatization on the plate and precise computation of the calibration line within an automatic measurement and evaluation, represents a new, inexpensive analysis system. There are only 2 min of labour time (one fifth of that required in thin-layer chromatography) required for one sample and the total analysis time varies from 3 to 9 min (one third to one quarter of that required in thin-layer chromatography) based on one plate with 12 samples. The 95% confidence limits (N = 10) range between 0.5 and 3.0%.


Altered metabolism of androgens in elderly men with benign prostatic hyperplasia. Kinetics of testosterone, dihydrotestosterone (DHT) and 5alpha-androstane-3alpha,17beta-diol (3alpha-diol) were studied in 7 elderly healthy men (ages 61 to 80 years) with benign prostatic hyperplasia (BPH). Clearance rates were determined by the constant infusion technique with labeled testosterone and DHT. Metabolic clearance rate (MCR), conversion ratio (CR), the transfer constants (rho) and production rates (PB) were calculated. Plasma androgens were measured by specific radioimmunoassay. Plasma testosterone was 516 +/- 314 (SD) ng/dl, plasma DHT was 74.6 +/- 19.6 (SD) ng/dl and plasma 3alpha-diol was 16.4 +/- 4.1 (SD) ng/dl. An elevated DHT level in elderly men with BPH wasconfirmed. MCRT was 620 +/- 65 (SD) liter/day and MCRDHT was 393 +/- 50 (SD) liter/day. Both MCRT and MCRDHT in elderly men were significantly lower than in young men. PBT was 3.2 +/- 2.1 (SD) mg/day and PBDHT was 291 +/- 87 (SD)migrogram/day. PBDHT was the same in elderly and young men. DHT production is maintained in elderly men despite reduction of testosterone production. From the data, it was claculated that in contrast to young men where greater than 80% of blood DHT is from secreted testosterone, over 50% in elderly men is derived from secretion or production of DHT by the testis or even more likely the prostate.


Simplified silver-plating stain for flagella. Rhodes' silver-plating technique for staining flagella was tested for its reliability and convenience as a routine procedure in the clinical laboratory. Modifications were made in the stain preparation and the procedure of staining and were tested with smears of known motile gram-negative nonfermentative bacilli. The stain has proved to be accurate and reliable and can be easily utilized with a minimum of training.


Fluorescent-antibody studies on selected strains of Bacteroides fragilis subspecies fragilis. Antisera against seven strains of Bacteroides fragilis subspecies fragilis were produced from dense suspensions of whole cells. These sera exhibited high agglutination titers with homologous antigens. Reciprocal cross-reactions in agglutination tests with each immunizing strain yielded lower titers. Both the indirect and direct fluorescent-antibody techniques were used to evaluate these reagents in the serological identification of 24 defined strains of B. fragilis subspecies fragilis. Subspecies and even strain specificities were noted with particular antisera. A pooled antiserum and conjugate were prepared and studied. Study results showed that specific and high-titered antisera against strains within this subspecies can be produced by the methods described herein and that possibly more than one serotype exists within the seven strains studied. The development of more antibody pools will be necessary to encompass a wider antigenic coverage before the fluorescent-antibody technique can be relied upon altogether for serologically identifying isolates of B. fragilis subspecies fragilis. Test data showed that the indirect method of fluorescent-antibody staining with whole antiserum is an excellent means of identifying strains of this organism.


Histopathological occurrence and characterisation of calcium oxalate: a review. Oxalosis is the histological manifestation of a number of diverse clinicopathological states involving abnormalities of both endogenous and exogenous oxalate. Crystalline deposits of calcium oxalate, usually first detected by their birefringence, may be characterised by a combination of their physical and tinctorial properties.


The olivocerebellar projection studied with the method of retrograde axonal transport of horseradish peroxidase. V. The projections to the flocculonodular lobe and the paraflocculus in the rabbit. HRP was injected in the flocculonodular lobe and the paraflocculus in the rabbit to determine the areas of the inferior olive which project onto these cerebellar regions. Following injections in the flocculus labeled cells occurred in the dorsal cap and the rostralmost tip of the medial accessory olive. Following injections in the nodulus labeled cells were likewise found in the dorsal cap, but in addition in the rostralmost part of the dorsomedial cell column and the adjoining part of the medial accessory olive. Injections in the dorsal paraflocculus gave rise to labeling in the rostrolateral part of the medial accessory olive, while injections in the ventral paraflocculus resulted in labeling in the principal olive, mainly in the lateral part of the ventral lamella. Injections in the lateral third of the dentate nucleus gave rise to labeling mainly in the dorsal lamella of the principal olive. The results are discussed with reference to those obtained by previous authors. There are both similarities and discrepancies. It appears from what is known of afferents from areas mediating visual impulses to the inferior olive that the olivary areas projecting onto the flocculonodular lobe, and possibly the dorsal paraflocculus, may mediate visual impulses to these lobules.


The olivocerebellar projection in the cat studied with the method of retrograde axonal transport of horseradish peroxidase. VI. The projection onto longitudinal zones of the paramedian lobule. Microinjections (30-50 nl) of a horseradish peroxidase (HRP) suspension of 25% (wt./vol.) were made in different folia of the paramedian lobule of cats, and the sites of occurrence of labeled cells in the inferior olive were precisely determined. In each case only a small number of cells are labeled, aggregated in a minute area. The labeled cells are found within three only of the four olivary areas previously determined (Brodal et al., '75) to project onto the paramedian lobule (fig. 1): one area in the rostral half of the medial accessory olive, another in the dorsal accessory olive (except its caudalmost part), and a third in part of the caudal half of the dorsal lamella of the principal olive. Labeled cells were never found in the fourth area, the ventral lamella. A distinct zonal pattern in the projection is demonstrated (figs. 3, 5B): a middle longitudinal zone of the paramedian lobule receives olivary afferents from the area in the medial accessory olive, a medial zone from part of the projection area in the dorsal accessory olive, a lateral zone from part of the projection area in the dorsal lamella. This zonal projection appears to extend throughout the length of the paramedian lobule (the two caudalmost folia could not be studied). tthe somatotopical pattern in the projections from the accessory olives described previously (Brodal et al., '75) is confirmed. The pattern of a zonal projection obtained with the HRP-method (fig 5B) is simpler than that deduced by Armstrong et al ('74) from recordings of antidromic potentials in the olive (fig 5A). Concerning main points there is satisfactory agreement. The phenomenon that following microinjections of HRP in superficial parts of the folia labeled cells occur within parts only of the regions of the olive which contain labeled cells following large HRP-injections in the paramedian lobule is discussed.


Vitamin D binding factors in bovine blood. Both 25 [25,26-hydrogen-3] hydroxycholecalciferol and [1alpha,2alpha-hydrogen-3] cholecalciferol were added to bovine plasma in vitro. Analysis by gel-filtration and ion exchange chromatography, electrophoresis, ultracentrifugation, competitive binding specificity studies, and plasma stripping showed that vitamin D circulated with a protein of alpha-globulin mobility. This globulin had a much higher affinity for 25-hydroxycholecalciferol while vitamin D3 appeared to be associated first with an alpha-lipoprotein and with time because associated with the alpha-globulin. This alpha-globulin had a molecular weight of approximately 70,000 as determined by gel-filtration. Cholecalciferol appeared to bind tightly to the alpha-lipoprotein and resisted being stripped from the plasma. Thus, alpha-globulin appears to be the major carrier of vitamin D in the blood while the alpha-lipoprotein may aid in the transfer of cholecalciferol from the gut to the liver via the lymph system.


Complement-mediated release of histamine from human basophils. III. Possible regulatory role of microtubules and microfilaments. The release of histamine by normal human leukocytes (basophils) following in vitro challenge with activated complement (zymosan-treated serum) was previously reported. In this study, the effects of various pharmacologic agents on this release mechanism were compared with allergen-induced release of histamine. Colchicine and vinblastine antagonize the polymerization of tubulin to form microtubules, and both agents inhibited complement-and allergen-triggered release of histamine from basophils. Finally, treatment with cytochalasin B, a fungal product known to interfere with microfilament formatin, resulted in enhanced release of histamine from complement-treated basophils but no significant change in the percentage of histamine released from allergen-treated basophils. These findings suggest that microtubules and/or microfilaments are involved in complement-induced secretion of histamine by human basophils.


Immuno-electron-cytochemical localization of the somatostatin cells in the human antral mucosa. Immuno-cytochemical methods were used to identify, in light and electron microscopy, the somatostatin-containing cells of the human antral mucosa. By means of immunoperoxidase and immunofluorescence methods sequentially applied on the same section, it was shown that the somatostatin cells are distinct from the gastrin cell population; these two endocrine cell types are often closely related. On ultrathin sections from aldehyde-fixed. Epon-araldite embedded tissues, the site of storage of somatostatin was localized with the peroxidaseantiperoxidase complexes technique, after removal of the resin by means of sodium ethoxide. This procedure represents a new technical approach to the use of electron-cytochemical techniques. The results indicate that somatostatin, a growth hormone release inhibiting factor, is localized in the endocrine granules of the D cells.


Immunocytochemical localization of a folicle stimulating hormone-like molecule in the testis. A follicle-stimulating hormone (FSH)-like molecule was localized in normal adult rat testes as well as testosterone-treated hypophysectomized rat tests with an unlabeled antibody (anti-FSH), peroxidase-antiperoxidase complex technique. Anti-FSH bound specifically to ultrathin sections of acrosomes of spermatids and intranuclear bodies of early spermatids. Quantitation of staining intensity demonstrated that FSH, used as an absorbing antigen, would significantly reduce this binding. There was less anti-FSH binding to the acrosomes of spermatozoa in the body and tail of the epididymis as compared to the less mature germ cells located in the testis and head of the epididymis. The acrosomal and nuclear staining of spermatids taken from hypophysectomized animals was similar to staining observed in sham injected animals. Taken together, these results suggest that there is a molecule within the acrosome that is immunologically similar to FSH. Most importantly, these results emphasize the importance of conducting physiologic experiments in conjunction with immunocytochemical studies.


Quantitation of cellular deoxyribonucleic acid by flow microfluorometry. This report characterizes for the first time an easy, reproducible means of standardizing the relative fluorescent units normally reported for flow microfluorometry. Absolute values for deoxyribonucleic acid/cell are obtained by using nucleated red blood cells as references. Cell were selected and characterized for the quantitative analysis of deoxyribonucleic acid per cell over a range from 2 pg/cell to 93 pg/cell using literature values for species having nucleated erythrocytes. Fluorescence staining by either acridine-orange (green wavelength) or propidium iodide (red wavelength) gave linear curves over the entire range investigated only when "gain controls" and current are optimized. The range was equivalent to mammalian cell values from 1 N (=3.5 pg deoxyribonucleic acid/cell) to 28 N (=91 pg deoxyribonucleic acid/cell). The standard curves obtained with nonmammalian erythrocytes were compared to mammalian free-cell preparations of bovine thymus and liver cells which fell at 6.8 and 6.9 pg deoxyribonucleic acid/cell, respectively. The routine use of these easily obtainable red blood cells will allow ready comparisons on the basis of absolute values for deoxyribonucleic acid per cell for work between experiments, work between staining procedures and dye types and work between laboratories.


Histologic and biochemical identification and characterization of an elastin in cartilage. Histochemical and chemical techniques have been used to identify, isolate and characterize elastin from certain bovine cartilages. The results strongly suggest that in addition to fibroblasts and smooth muscle cells, chondroblasts also synthesize elastin. Some of the possible functions of elastin in elastic cartilages are discussed and the possiblity of a new type of elastin perhaps unique to cartilage is suggested.


Retrieval of cryostat section for comparison of histochemistry and quantitative electron microscopy in a muscle fiber. A method is presented that can be used to perform histochemical and morphometric analyses on the same muscle fiber. Freshly dissected fibers from medial gastrocnemius muscle of adult guinea pig were kept at a resting length and rapidly frozen. Serial frozen cross-sections were cut and reacted for myofibrillar adenosine triphosphatase and succinic dehydrogenase. The adjacent section, while still frozen, was immersed into 20 degrees C glutaraldehyde fixative to which EGTA was added to minimize artifactious contraction. The fixed section was processed for electron microscopy and the section rotated before thin sectioning to give longitudinal sections enabling study of sarcomeres. Ultrastructure was well-preserved despite slight disorganization of the contractile filaments and some vesiculation of the sarcoplasmic reticulum. The Z line width was measured and the mitochondrial volume fraction estimated by point counting morphometry from 89 fibers. The fibers with dark myofibrillar adenosine triphosphatase staining have Z widths of 547 +/- 165 A (n=69) and thoshosphatase staining have Z widths of 547 +/- 165 A (n=69) and those with light stain have 1023 +/- 113 A (n=20). The density of the succinic dehydrogenase reaction product in the fibers was divided into dark and light and the mitochondrial volume fractions were foud to be 4.3 +/- 2.1% (n=52) and 1.0 +/- 1.1% (n=37), respectively.


Trypsin-like activity in the vaginal epithelial cells of the rat. Rat vaginal epithelial cells have trypsin-like activity as shown by the formation of a colored product when the cells are incubated with alpha-N-methyl alpha-N-toxyl-L-lysine beta-naphthol ester and hexazotized pararosanilin. This enzyme activity in vaginal smears is maximal at proestrus, i.e., the day in the 5-day estrus cycle when plasma estrogen is maximal. Only the rounded nucleated epithelial cells present at late diestrus, proestrus and early estrus demonstrate the trypsin-like enzyme activity. These are the cells that stain blue in the Papanicolaou method. Preincubation of cell suspensions with the serine protease inhibitor, p-nitrophenyl p-guanidino benzoate, prevented the enzyme staining reaction, further demonstrating the trypsin-like nature of the cellular enzyme. The advantages of this enzyme staining technique over the fibrin plate method for the demonstration of trypsin-like enzymes in cells are increased resolution and ability to show trypsin inhibitor effects.


Noncytotoxic IgE-mediated release of histamine and serotonin from murine mastocytoma cells. Cultured murine mastocytoma (AB-CBF1-MCT-1) cells were stimulated to release endogenous or incorporated histamine or serotonin by an IgE-mediated mechanisms without loss of viability. Stimulation was achieved by incubation of the cells with rat IgE-anti-IgE, rat IgE-anti-light chain, fluoresceinated rat IgE-anti-fluorescein, IgE-enriched mouse anti-ovalbumin-ovalbumin, or covalently linked dimers of rat IgE, at doses similar to those optimal for normal peritoneal mast cells. Active cell metabolism and Ca++ were required to obtain release. Despite the latter, no dose of the calcium ionophore, A23187, could be found which caused release without concomitant cytotoxicity. Phosphatidylserine did not enhance release.


Kinetics of the in vitro lymphocyte response to myelin basic protein in the the Lewis and Brown Norway strains of rat. Several parameters of the in vitro lymphocyte proliferative response to myelin basic protein (BP) in Lewis (Le) and Brown Norway (BN) rats were examined. The results demonstrate that BN rats, a strain normally resistant to BP-induced experimental allergic encephalomyelitis, and Le rats, a strain readily susceptible to the disease, have similar patterns of the proliferative response to BP. An important difference, however, is that BN lymphocytes, although responding significantly to BP, are unable to proliferate to the same level as Le lymphocytes. In experiments measuring the lymphocyte response as a function of antigenic stimulus, days of culture, or type of adjuvant used, the BN rat peak response was in general 70% or less of the Le rat peak response. Furthermore, the BN lymphocyte response was reduced when B cells were removed whereas there was no effect in the Le rat. A negative feedback mechanism, possibly suppressor cells, has been suggested to explain these differences.


Suppressor T cells: presence in mice rendered tolerant by neonatal treatment with anti-receptor antibody or antigen. Specific tolerance to phosphorylcholine (PC) was induced in BALB/c mice by two methods. Neonatal mice received a single injection of either: 1)PnC, the C-polysaccharide from S. pneumoniae, R36a vaccine which has PC as a major antigenic determinant or 2) ARA, an homologous antibody directed against the receptor for PC. Spleen cells from animals treated as neonates  with either PnC or ARA were specifically suppressed for the response to PC antigens in vitro. In addition, cells from either group of unresponsive animals co-cultured with spleen cells of normal BALB/c mice markedly suppressed the response of the normal cells to PC. Greater than 90% inhibition of the plaque-forming cell response was obtained when unresponsive cells were mixed with normal cells in ratios of 1:1 or greater. Equal numbers of cells from animals made unresponsive by PnC or ARA produced an equivalent degree of suppression. Neither supernatants of cultures nor sera of animals of either unresponsive group suppressed the response of normal spleen cells to PC. Suppression by cells from both groups of tolerant mice was eliminated by treatment with anti-Thy 1.2 serum and C. Presumably, a common cell is responsible for suppression caused by cells from mice made unresponsive by either procedure.


Idiotypes on galactan binding myeloma proteins and anti-galactan antibodies in mice. Antibodies with specificity for beta1,6 linked D-galactoses were induced in mice by immunization with gum ghatti. Idiotypic antisera were prepared in rabbits and mice by immunization with 8 BALB/c IgA(k), beta1,6D-galactan binding myeloma proteins (beta6GALBMP). Two kinds of idiotypic sera were obtained: cross-specific sera that reacted with two or more beta6GALBMP but not other BALB/c myeloma proteins, and individual idiotypic sera that reacted with only the beta6GALBMP used in the immunization. Antibodies with specificity for beta1,6 linked D-galactans shared cross-specific idiotypes with beta6GALBMP. Only one of seven individual idiotypes associated with beta6GALBMP was found on galactan antibodies. Since all beta6GALBMP thus far have the same Vk and VH isotope composition the results indicate an extensive heterogeneity among galactan-binding immunoglobulins in BALB/c mice. It is speculated that some of this diversity may arise from somatic rather than germ line gene mutations.


Functional differentiation of B lymphocytes in congenital agammaglobulinemia. II. Immunochemical analysis of the in vitro primary immune response. Cultures of peripheral blood lymphocytes (PBL) in which specific hemolytic plaque-forming cells (HcPFC) had been induced were labeled with 14C-amino acids. Antigen-specific products in the culture supernatants were characterized by using indirect immune precipitation in conjunction with specific immunoabsorbents and/or gel filtration followed by SDS-polyacrylamide gel electrophoresis. After 5 days of culture with antigen (sheep red blood cells or ovalbumin) newly synthesized IgM and specific IgM antibody were demonstrated in culture supernatants from normal donors and from four out of five patients with congenital agammaglobulinemia (cAgamma). Secreted products bound specifically to antigen and pretreatment of labeled supernatants with anti-mu and anti-L chain antisera, but not with anti-gamma antiserum, prevented binding. Typical mu- and L chains constituted only a proportion of the anigen-binding peptides recognized by the anti-mu reagents. Induction of IgM antibody synthesis was dependent on the presence of antigen and was correlated with the generation of HcPFC. No major differences between the antigen-induced products of cAgamma and normal PBL were observed. These findings suggest that in the absence of terminal B cell differentiation in vivo, certain patients with cAgamma possess precursor cells that can respond to antigen in vitro with the synthesis of specific humoral products, including IgM antibody.


A cross-reaction between beta2-microglobulin and kappa-light chains. Certain antisera to immunoglobulins containing kappa-chains show the presence of antibodies that cross-react with beta2-microglobulin. This was most apparent with an antiserum made to highly purified F(ab) fragments of Fr II gamma-globulin. These cross-reactive antibodies caused positive fluorescence and cytotoxicity reactions with a variety of cell types including T cells. These reactions were completely removed by absorption with highly purified kappa-chains but not with lambda-chains or lambda immunoglobulins. beta2-microglobulin preparations also absorbed or inhibited the special cellular reactivities. Evidence was obtained that HLA-bound beta2-microglobulin was more efficient in this respect. The possibility is discussed that similar cross-reactive antibodies may have been involved in some previous studies of inhibition of T cell function by immunoglobulin antisera.


Requirements for inducing tolerance of hapten-specific delayed hypersensitivity: epitope density. Tolerance to hapten-specific antibody formation and delayed hypersensitivity was examined in adult rabbits. The azobenzenearsonate (ABA) or sulfonate-specific antibody response to hapten-hemocyanin immunogens was suppressed by deaggregated hapten-rabbit IgG conjugates given 21 and 14 days before challenge. High affinity antibody was preferentially suppressed. Delayed hypersensitivity to ABA-tyrosine was suppressed by deaggregated ABA-rabbit IgG conjugates injected 17 and 10 days before challenge. Conjugates with a high hapten density, ABA15-23-rabbit IgG were effective tolerogens. Conjugates with four to six ABA groups per carrier molecule were very poor tolerogens. Increasing the amount of low substituted conjugate injected did not improve tolerogenicity. It appears that a high epitope density is required for effective induction of tolerance to ABA-specific delayed hypersensitivity in the rabbit.


Immune responses to vaginal or systemic infection of BALB/c mice with herpes simplex virus type 2. The temporal relationships among the humoral and cellular immune responses were defined in BALB/c mice after vaginal or systemic infection with herpes simplex virus type 2 (HSV-2). After vaginal infection, mice showed evidence of clinical vaginitis on days 4 to 6 and HSV-2 replication was detected locally in the vaginal secretions, cervix, vagina, and uterus before the virus subsequently spread to the central nervous system. Death from encephalitis occurred between 7 and 10 days after infection. Vaginal infection was associated with significant delayed type hypersensitivity and splenic proliferative cell-mediated immune responses which appeared during the acute infection and waned by 3 weeks. There was almost no evidence of a systemic neutralizing antibody response at any time after vaginal infection. In contrast to the local vaginal infection, systemic i.v. HSV-2 infection induced a humoral response as well as the two cellular immune responses. Although both cellular immune responses appeared during the acute infection (days 6 to 14) and persisted for approximately 5 weeks, the humoral response appeared in surviving animals and persisted for at least 4 months. Thus, vaginal HSV-2 infection was associated primarily with transient cellular immune responses, whereas i.v. HSV-2 infection induced prolonged systemic humoral and cellular immune responses.


B cell tolerance. I. Analysis of hapten-specific unresponsiveness induced in vitro in adult and neonatal murine spleen cell populations. The cellular mechanisms and tolerogen dose requirements of hapten-specific unresponsiveness induced in vitro by using 2,4,6-trinitrophenyl human gamma-globulin (TNP17HgG) were analyzed in adult and neonatal murine splenocytes. Tolerance induction in both cell populations was found to be independent of non-B cell effects including BAtheta-positive cells, Ly 2.2-positive cells, adding or reducing the number of macrophages, and large excesses of HgG. The tolerance induced was specific and not "infectious", further excluding a role for suppressor T cells. Neonatal splenic B cells were rendered tolerant by doses of TNP17HgG 1000-fold less than those required to produce similar tolerance in splenic B cells from adults. These findings support the concept of functional clonal abortion as a mechanism for producing tolerance to self antigens.


Mechanism of histamine release by formyl methionine-containing peptides. Small, acylated, methionine-containing peptides release histamine from human basophils. The characteristics of this reaction were compared to that of C5a- and IgE-induced release. fMet peptide-induced release requires Ca++ and is inhibited by EDTA in a manner similar to IgE- and C5a-mediated reactions. The fMet-Phe-Met-initiated reaction is complete within 2 min at temperatures of 25, 30, and 37 degrees C; but does not occur at 0 degrees C. There was a large variation in the capacity of leukocytes from different donors to release histamine with fMet peptides. However, there was no correlation in the capacity of leukocytes to release histamine with fMet-Phe-Met and their release with C5a or anti-IgE. The release by fMet-Phe-Met (but not by C5a or anti-IgE) was reversibly inhibited by a nonreleasing tripeptide. Leukocytes could be desensitized to the action of active fMet-peptide by preincubation with the peptide in the absence of cations. After washing, these cells released normally with C5a or anti-IgE. Conversely, cells desensitized to the action of C5a- or IgE-mediated reactions released normally with fMet peptides. There was cross-desensitization between different active peptides, and inactive peptides could not desensitize the leukocytes. Pharmacologic agents had similar effects on C5a and fMet peptide-induced release (e.g., lack of enhancement with deuterium oxide; enhancement with cytochalasin B; and inhibition with aminophylline and dibutyryl cyclic AMP). Therefore, histamine release with fMet peptides is initiated by their binding to and activation of a specific receptor on the basophil; the reaction beyond that point is similar to the C5a-mediated reaction.


A restaining method to restore fluorescence in faded preparations of tissues treated with the indirect immunofluorescence technique. We report a restainin method for restoring fluorescence in paraffin-embedded tissue sections previously treated with the indirect immunofluorescence technique. Antisera to gastrin and group II pepsinogens were used. Fluorescence was restored in completely faded sections retrived from storage files, as well as in sections that had faded partially either with exposure to fluorescence microscope illumination or after counterstaining with hematoxylin and eosin.


An improved stain for immunodiffusion tests. We describe an improved immunodiffusion stain using trichloroacetic acid to mordant dying by Crocein scarlet. Background is readily cleared without over-destaining using 0.3% acetic acid.


A micro-method for the study of in vitro lymphocyte transformation to specific antigens. A quantitative method for studying blast-transformation of cultured lymphocytes in the presence of specific antigens is described. The technique is performed in 0.3 ml microtrays, using 2.5 X 10(5) lymphocytes per culture, with an optimal culture period of 6 days. Cultures are pulsed for 4 h with [3H]-thymidine, giving maximum incorporation and minimum cellular damage. The study also describes some disadvantages of using lymphocyte responses to phytohaemagglutinin (PHA), as a marker of T-cell function. Experiments to establish ideal conditions for each stage of the method are described. The technique can be adjusted to accommodate different antigens, cell types and for the study of animal as well as human lymphocytes.


Experimental autoimmune encephalomyelitis in mice: antigen binding lymphocytes to basic protein of myelin in susceptible and resistant strains. Antigen-binding lymphocytes (ABL) to basic protein of myelin (BPM) were demonstrable in the thymus and spleen of various strains of mice. Counts of ABL did not differ in strains resistant and susceptible to EAE, and did not differ when human or murine 125I-labelled BPM was used as a labelled antigen to detect ABL. After injection with mouse spinal cord and adjuvants, under conditions known to be immunogenic and encephalitogenic for susceptible strains of mice, counts of ABL increased two- to four-fold in suceptible strains but did not icnrease in resistant strains. Antigen binding could not be accounted for by the attachment to cells of cytophilic antibody with specificity for BPM, because such antibody was seldom demonstrable even after immunization. Inferences from the study were: (i) that tolerate (non-responsiveness) to certain autoantigens at least must depend on control over continually present antigen-reactive lymphocytes rather than on any lack of capacity to recognize antigen; (ii) differences in susceptible and resistant strains are evident after, rather than before, immunization.


Perinatal immunity to group B beta-hemolytic streptococcus type Ia. Sera from 14 of 56 adult women protected mice from intraperitoneal challenge with mouse-virulent group B Streptococcus serotype Ia, and sera from seven of 25 nonparturient women in this group were bactericidal for greater than 99% of the organisms in the presence of normal polymorphonuclear leukocytes. There were no discrepancies between the in vivo and in vitro assays. Protective activity was found in the IgG class in seven sera, and in the IgM class in one. Opsonic activity was partially dependent on heat-labile serum factors. Of 31 mother-cord serum pairs studied, seven maternal sera were protective, but four of the corresponding cord sera were not. Pooled human gamma-globulin injected by either the intraperitoneal or the intramuscular route protected mice from bacterial challenge.


Structural subunits of poliovirus particles by electron microscopy. Electron microscopy of poliovirus particles and empty capsids under various conditions of specimen deposition and staining visualizes the dissociation products of these particles. The dissociation proceeds in steps; it begins with the expansion of particles and leads to the final product of the dissociation--a cluster of several sub-particles of equal size (approx. 100 A in diam.). A scheme of the dissociation is proposed on the basis of the observed intermediates.


Inhibition of vesicular stomatitis virus by kethoxal bis (thiosemicarbazone). Kethoxal bis (thiosemicarbazone) (KTS) inhibited replication of, and plaque formation by, vesicular stomatitis virus (VSV) in chick embryo cells. No other thiosemicarbazones tested were effective. Virus-specific m-RNA and protein synthesis were inhibited by KTS. However, virion RNA-dependent RNA synthesis was not inhibited by the drug. Treatment of VSV virions directly with KTS produced enhancement, rather than inactivation, of plaque formation. KTS inhibited cellular DNA and RNA synthesis by 67 and 25% respectively. Since cellular DNA and RNA synthesis are not required for VSV replication, the inhibition of these processes is probably unrelated to the antivirial activity of KTS. Cellular protein synthesis was inhibited 24% by KTS. Unexpectedly, synthesis of four proteins was induced in KTS-treated uninfected cells.


Mortality of patients with Parkinson's disease treated with levodopa. The effect of levodopa on the mortality of patients with Parkinson's disease was investigated in 349 patients treated with levodopa or levodopa combined with decarboxylase inhibitor during 1969-1975 inclusive. During the study period, 61 patients died. The expected mortality was 32.99 resulting in a ratio of actual to expected deaths of 1.85. The excess mortality was accounted for by patients with a severe disease at entry and especially, by the less favorable effect of levodopa treatment than in the living patients. In comparison with the prelevodopa era, the reduction of mortality and the increase of life expectancy of patients with Parkinson's disease during levodopa treatment possibly reflect the decrease of the early mortality due to Parkinson's disease.


The bioavailability of phenytoin.

The bioavailability of phenytoin. The bioavailability of three commercial products of phenytoin (Epanutin, sodium salt; Phenhydan, calcium salt; Zentropil, free acid) was studied relative to a standard solution of sodium phenytoin. Each preparation was given for 14 days in a daily dose of 300 mg according to a cross over design. The three brands tested were equivalent as to their bioavailability with respect to plasma concentrations during one dosing interval, the area under the plasma level time curve, the time of peak plasma concentration, and the urinary excretion of the primary metabolite. Therefore, these phenytoin brands are bioequivalent and exchangeable for antiepileptic treatment. The standard solution of sodium phenytoin had a higher bioavailability as compared to the commercial products.


Familial focal loss of cross striations. Two patients, a brother and sister, both suffering from congenital generalized muscle weakness with a progressive course are reported. Muscle biopsy revealed areas with loss of cross striations in the muscle fibers, electronmicroscopically presenting complete disorganization of the myofibrils with streaming of the Z discs and absence of mitochondria. Vesicular nuclei with prominent nucleoli were present in these areas. There was a type I fiber predominance in both cases. The mean diameter of the type I muscle fibers in one of the cases was too small. Motor endplate alterations in this patient gave no evidence of denervation but were suggestive of a delayed development of motor nerves.


Temperature dependence of normal sensory nerve action potentials. Sensory conduction velocities of normal subjects are increasing linearly with rising temperature. The duration of the compound sensory action potentials recorded from the median nerve at the wrist and elbow shows a negative temperature coefficient. The peak-to-peak amplitude of these potentials increases from 22 degrees to approximately 26 degrees C and then decreases progressively again up to 36 degrees C. It is believed that this behavior is due to a combination of decreasing temporal dispersion, height and duration of the individual spike potentials.


Changes of sensory conduction velocity and refractory periods with decreasing tissue temperature in man. Changes with temperature of maximum sensory nerve conduction velocity as well as absolute and relative refractory periods were tested in 14 human subjects. Corresponding to previously published findings maximum conduction velocity decreased with cooling following a Q10 of +1.4. The absolute and relative refractory periods were increased by cooling, the Q10 being -3.1 and -3.35 respectively. There was a tendency showing a more pronounced temperature effect at low temperatures. The Q10 and the steepness of the regressionline changed at the level of 26.9 degrees C, but were significant for the relative refractory period only.


A temporal study of survival of patients with pontine gliomas. Twenty-four cases of pontine glioma were treated over a 16 year period. Survival times are discussed, particularly long survival times, on the basis of 13 cases autopsied. Onset occurred in an age range of 5 to 60 years, and 5 of the 13 autopsied cases involved children. The average survival time was 9 months except for 2 long survival cases, one of 4 years and 7 months and the other of 14 years and 10 months. The longer the survival time, the greater was the number of neurological symptoms detected, but there was no relationship between the involvement of cranial nerves and the survival time. The improvement of cranial nerve disorders was more prominent in the long survival cases than that of other neurological disturbances. The time from onset of symptoms to admission was longer for long survival cases than the others, and the autopsies of two long survival cases revealed astrocytoma. There were no cases which survived more than one year in the glioblastoma multiforme group.


Biochemical findings in multiple sclerosis. V. Transformation of lymphocytes from patients with multiple sclerosis by human basic protein. Lymphocyte stimulation tests with human basic protein of myelin were performed on patients with multiple sclerosis, with other neurological diseases and on normal subjects. In both MS and OND group, a hypersensitization to basic protein was seen in about one third of the cases. All normal subjects, except one, had negative responses. In the MS group, a positive correlation could be found with some features of the disease: significantly more positive responses were found in independent patients with a short duration of illness and in those with an oligoclonal distribution in the CSF. The authors compare their results with those of the literature. The possible role of BP in pathogeny of MS and OND is discussed.


Amyloid neuropathy due to monoclonal gammopathy. A case report. A case with peripheral neuropathy due to monoclonal gammopathy is reported. There was striking, dissociated symmetrical sensory loss and paresis, the legs being affected predominantly. The case is compared with the few reported cases of myelomatous neuropathy, and the clinical and histological findings are discussed in view of the pathogenesis.


Malignant diseases among patients with multiple sclerosis. The frequency of malignant diseases among 1866 living and 340 deceased multiple sclerosis (MS) patients was investigated in Finland. The study revealed a low prevalence (0.64%) and mortality (0.07%) rate of cancer among MS patients. The difference between MS patients and general population was significant. The highest rate was found in the group from 40 to 49 years while in the general population the rate of cancer among MS patients tended to fall after the age of 50. The possible role of selenium, one of the antioxidants in the pathogenesis of MS and cancer, is discussed because recent data have shown a very high negative correlation between selenium and cancer death rates.


Lectin and human blood group determinants of Schistosoma mansoni: alteration following in vitro transformation of miracidium to mother sporocyst. A mixed agglutination assay method was employed to detect the presence of surface determinants for various lectins and human blood group antibodies on Schistosoma mansoni miracidia and cultured mother sporocysts. Miracidia were found to possess surface receptors for the lectins Con A (concanavalin A), anti-Heel (eel serum agglutinin), and anti-ADb (Dolichos seed extract), as well as human anti-A antibodies. Following in vitro transformation of the miracidium to mother sporocyst, anti-Heel and human anti-A receptors were no longer detectable on the sporocyst surface, while determinants for Con A and anti-ADb remained essentially unaltered. It is concluded that transition of the miracidium to the sporocyst results in the alteration of surface molecular structures on schistosome larve. Furthermore, since determinants for Con A, anti-Heel, anti ADb, and human anti-A have been found associated with macromolecules in the hemolymph of the snail Biomphalaria glabrata (Stnislawski et al., 1976), there is now evidence that miracidia and mother sporocysts of S. mansoni and their snail host share molecules with common lectin and human blood group determinants.


Survival in bilateral Wilms' tumor--review of 30 National Wilms' Tumor Study cases. Bilateral tumors were found in 33 patients or 5.4% of 606 children enrolled in the National Wilms' Tumor Study. Twenty six of 30 patients (87%) for whom detailed clinical information is available survived 2 yr after conservative surgery and radiation therapy. All patients were given one or more chemotherapeutic agents. A variety of surgical procedures were used including biopsy only, nephrectomy, and partial nephrectomies. Radiation therapy doses ranged widely, but between 1000 and 2000 rad apparently sufficed for local control in most cases. Recommendations regarding management by surgery, radiation therapy, and chemotherapy are given.


The relationship between histamine secretion and 45calcium uptake by mast cells. 1. Unstimulated mast cells from the peritoneal cavity of the rat take up (45)Ca: the initial phase of rapid uptake being complete after 1 min incubation of the cells with the isotope. Stimulation of the mast cells with an antigen-antibody reaction, dextran or concanavalin A induces an increase in the uptake of (45)Ca which is accompanied by a release of granular material: this increase in (45)Ca uptake is also complete in 1 min. The majority of the stimulated (45)Ca uptake cannot be explained in terms of binding of Ca to released granular material, or to an enlargement in either the extracellular compartment or the cell surface area.2. The magnitude of the increase in (45)Ca uptake caused by stimulating the mast cells increases when the degree of histamine secretion increases.3. The increased (45)Ca uptake induced by stimulation of the mast cells and the degree of histamine secretion are both dependent on extracellular H ion concentration. Changes of pH cause similar changes in (45)Ca uptake and secretion with maxima at pH 7.5.4. Two thirds of the (45)Ca uptake induced by an antigen-antibody reaction or by the Ca ionophore A 23187 is unaffected by inhibiting glycolysis and oxidative phosphorylation. Histamine secretion on the other hand is practically abolished by this metabolic inhibition. Thus, (45)Ca uptake proceeds in the absence of the discharge of granules.5. Dibutyryl cyclic AMP or theophylline inhibit both the increase in (45)Ca uptake and the histamine secretion caused by stimulating mast cells with an antigen-antibody reaction. Cyclic AMP, cyclic GMP and dibutyryl cyclic GMP have no effect on uptake or secretion.6. The Ca ionophore, A 23187, induces uptake of (45)Ca and histamine secretion, neither effect being inhibited by either dibutyryl cyclic AMP or theophylline.7. Phosphatidyl serine increases both (45)Ca uptake and the histamine release induced by an antigen-antibody reaction, dextran or concanavalin A.


Spontaneous histamine secretion from mast cells in the presence of strontium. 1. Histamine secretion from rat peritoneal mast cells occurs spontaneously in the absence of an external stimulus. Spontaneous secretion increases as the concentration of Sr in the extracellular medium is raised from 1 to 10 m-mole/l. Ca 0.1-10 m-mole/l. does not increase spontaneous secretion.2. Spontaneous histamine secretion in the presence of Sr occurs slowly compared with evoked histamine secretion, reaching a maximum only after more than 120 min incubation with Sr 10 m-mole/l. at 37 degrees C and pH 7.6. Phosphatidyl serine, 10 mug/ml., increases the rate of spontaneous secretion in the presence of Sr.3. The spontaneous secretion occurring in the presence of Sr is highly dependent on the extracellular H ion concentration. Maximal secretion occurs at pH 8.4 and only a very limited secretion is detected at pH below 7.6. The rate of spontaneous secretion is also greater at higher pH. Inhibition of secretion caused by lowering the pH can be reversed by raising the Sr ion concentration over a limited range.4. Intact glycolytic and oxidative metabolism is required for the spontaneous secretion of histamine in the presence of Sr. Removal of extracellular glucose inhibits the secretion by about 80%, and the further addition of inhibitors of oxidative phosphorylation almost abolishes the secretion.5. Ca, Mg and Mn all inhibit the spontaneous secretion of histamine which occurs in the presence of Sr. The antagonism of the effect of Sr by Mg appears not to be competitive.6. Dibutyryl cyclic AMP, 10 mumole/l. to 3 m-mole/l. and theophylline, 30 mumole/l. inhibit spontaneous secretion in the presence of Sr. Cyclic AMP, AMP, and cyclic GMP 10 m-mole/l. are without effect on the spontaneous secretion. The inhibitory effect of dibutyryl cyclic AMP and of theophylline are dependent on pH: greater inhibition being achieved at lower pH.


Movement of strontium ions into mast cells and its relationship to the secretory response. 1. Non-stimulated mast cells take up (89)Sr and (45)Ca. There is a rapid phase of uptake of both labels which is complete in 1 min and the (89)Sr uptake is similar in magnitude to the (45)Ca uptake. A further slower phase of uptake occurs in the period from 1 to 30 min of incubation. The magnitude of this slower phase is about 6 times greater for (89)Sr than for (45)Ca.2. Non-stimulated and antigen-stimulated mast cells accumulate Sr which can be measured by atomic absorption spectroscopy. There is a direct relationship between Sr accumulation and histamine secretion which is independent of whether or not the cells are stimulated. 10% histamine secretion is associated with Sr accumulation of 0.25 f-mole/cell.3. The time course of (89)Sr uptake is similar to the time course of histamine secretion in non-stimulated cells.4. The uptake of (89)Sr is linearly related to the external Sr concentration in the range 0.5-16 m-mole/l. for both stimulated and non-stimulated cells.5. Ca, 10-1000 mumole/l. inhibits (89)Sr uptake in non-stimulated cells.6. Inhibition of glycolysis and oxidative phosphorylation prevents spontaneous histamine secretion in the presence of Sr without blocking the accumulation of Sr by the cells.7. Dibutyryl cyclic AMP, 10 m-mole/l. produces only 18% inhibition of (89)Sr uptake whereas histamine secretion is inhibited by 45% by the same concentration. The antigen-stimulated Sr uptake on the other hand can be completely inhibited by dibutyryl cyclic AMP, 10 m-mole/l.8. The uptakes of (89)Sr and Sr by unstimulated mast cells after 60 min incubation are dependent on extracellular H ion concentration. Both uptakes increase with increasing pH over the range pH 7-8.5.


What are anaerobic cocci?

What are anaerobic cocci? Criteria are suggested for the definition of "anaerobic cocci" as a general term to include all obligately anaerobic cocci. Such a definition draws a clear distinction between obligately anaerobic and micro-aerophilic strains and might form a basis both for characterisation of these organisms in the diagnostic laboratory and for further studies on their taxonomy and pathogenicity.


Microscopy of the junctional region between human coronal primary and secondary dentine. The juction between human primary dentine and regular and irregular secondary dentine was examined with a number of different light and electron microscopic techniques. In decalcified material, a narrow band along the innermost surface of the primary dentine stained intensely. The walls of the tubules within the band stained intensely, whereas the tubular walls within the bulk of the primary dentine were not stained. Generally, the walls of the tubules in both types of secondary dentine were also preferentially stained. Although not readily apparent in ground sections, observations of thin sections revealed a dramatic reduction in the number of tubules in regular secondary dentine. Generally, the radiodensity of the intertubular matrix was the same in primary and secondary dentine and the intensely stained band was not seen radiographically. The pulpal ends of the tubules in primary dentine were often occluded with a material having the same radiodensity as peritubular matrix. Both patent and occluded tubules were seen in irregular secondary dentine. Scanning electron microscopy of acid-etched specimens of secondary dentine revealed that some tubules had irregular walls of highly mineralized matrix which was less acid-soluble then the peritubular matrix of primary dentine.


Interactions of murine leukemia virus core components: characterization of reverse transcriptase packaged in the absence of 70S genomic RNA. Virions produced by cells in the presence of actinomycin D (Act D virions) contain reverse transcriptase but are deficient in 70S genomic RNA. To assess the role of genomic RNA in encapsidation of a functional reverse transcriptase and to study the interaction of the enzyme and its template in the cores of intact virions, the reverse transcriptase enzymes of normal and Act D virions were compared. The enzymes were indistinguishable by column chromatography, sedimentation velocity, or template/primer preferences. In addition, these enzymes showed equal sensitivity to inactivation by antibodies directed against Rauscher murine leukemia virus DNA polymerase. The enzymes from Act D and normal virions had similar thermal decay rates and were both protected against heat denaturation by natural and synthetic template/primers. By these criteria, the DNA polymerase molecules synthesized and assembled into virions in the absence of genomic RNA are identical to those packaged under normal conditions. Additional studies designed to measure protection of reverse transcriptase by genomic RNA were carried out by comparing the thermal lability of the enzyme in intact Act D and normal virions. The thermal decay rate of reverse transcriptase in Act D virions was identical to that in control virions. In contrast to the lability of the virion-associated enzyme, however, genomic RNA in control virions was stable to heat treatment.


Type-specific antigenic determinants on the major external glycoprotein of high- and low-oncogenic murine mammary tumor viruses. We recently showed that the 52,000-dalton external glycoprotein (gp52) of the highly oncogenic mouse mammary tumor viruses (MMTVs) of RIII, GR, and C3H mice contains both type- and group-specific antigenic determinants. This was demonstrated by using a competition radioimmunoassay with 125I-externally labeled virions and antisera to the gp52 of MMTV from RIII mice (Proc. Natl. Acad. Sci. U.S.A. 74:3564-3568, 1977). We report here that we were able to distinguish between the gp52's of the high-oncogenic MMTV of C3H mice [MMTV(C3H)] and the low-oncogenic MMTV of that same mouse strain [MMTV(C3Hf)]. This was accomplished by use of a competition radioimmunoassay with 125I-externally labeled virions of MMTV(C3H) and antisera prepared against MMTV(C3H). A comparison of the intact virion and purified gp52 radioimmunoassays showed that MMTV type-specific differences were enhanced with the intact virion radioimmunoassay. These differences were further magnified with appropriately absorbed antisera. These findings thus allow an immunological distinction between the surface glycoproteins of a low-oncogenic endogenous and a high-oncogenic exogenous MMTV of the same mouse strain.


In vitro transcription of the avian retrovirus genome by the alpha form of the viral RNA-directed DNA polymerase. The nature of transcription of the avian retrovirus RNA genome by the alpha form of the viral RNA-directed DNA polymerase has been investigated. Transcription was most efficient when Mn2+ was provided as the divalent metal ion. The patterns of DNA transcription using 70S RNA, 35S RNA-tRNAtrp, or 35S RNA-oligo(dT)12-18 template-primer complexes by the alpha DNA polymerase were essentially identical to those obtained using the alphabeta form. The alpha DNA polymerase appears to be deficient in the synthesis of true duplex DNA but is able to synthesize hairpin-structured DNA initiated at the 5' terminus of the viral genome on the tRNAtrp primer molecule.


Alpha-fetoprotein and human chorionic gonadotropin in the management of testicular tumors. Alpha-fetoprotein and human chorionic gonadotropin levels were measured by radioimmunoassays in 260 patients with genitourinary diseases, including 93 patients with testicular tumors. Elevations of alpha-fetoprotein and human chorionic gonadotropin were associated only with non-seminomatous germ cell testicular tumors. Our 32-month experience with serial measurements of the 2 markers in patients with these tumors shows that alpha-fetoprotein and human chorionic gonadotropin must be determined and that together they serve as accurate and sensitive indicators of metastases and are helpful in determining the effectiveness of therapy. However, they have limited value in the differential diagnosis of scrotal masses.


Transurethral prostatic resections under local anesthesia. Standard transurethral prostatic resections were done on 18 patients with benign prostatic hyperplasia. Local anesthesia was used with standard 1 per cent lidocaine local infiltration of the prostate and topical instillation into the urethra. This anesthesia was supplemented in a few instances by modest doses of intravenous tranquilizers, which has proved to be a safe, simple and effective procedure.


Minocycline diffusion into benign prostatic hyperplasia. Minocycline, a broad-spectrum, highly lipid soluble tetracycline that has generated interest in the treatment of chronic prostatitis, was evaluated for its possible ability to be concentrated in benign prostatic hyperplasia. Drug levels in the prostate, plasma, fat, muscle and urine were measured in patients undergoing open prostatectomy after preoperative intravenous minocycline. The concentrations of the drug in the prostate and serum were close (4.16 versus 3.01 microgram. per gm.), while drug levels in striated muscle and fat were consistently lower (2.92 and 0.77 microgram. per gm). Higher preoperative doses of drug yielded higher tissue levels. Drug delivery closer temporally to the operation yielded higher serum and prostatic levels as opposed to striated muscle and fat, suggesting a rapid diffusion of the drug into benign prostatic hyperplasia.


Development of an epithelial tissue culture line from human prostatic adenocarcinoma. The development of a tissue culture line of human prostatic adenocarcinoma is described. The culture (HPC-36), derived from tumor tissue explants, is a pure epithelial culture with characteristics of neoplastic cells, including a large nuclear size relative to the amount of cytoplasm, multiple nucleoli within the nucleus, many mitotic figures, the formation of multinucleated giant cells and the loss of contact inhibition. The cells also stained positively for acid phosphatase and have been grown in monolayer and suspension cultures. The HPC-36 cells presently are being studied to determine whether they are true descendants of the cancer cells.


Use of heterogenous and monospecific antisera for the diagnosis of bladder cancer. A rabbit antibody to antigens present in urine from bladder cancer patients was prepared and used in conjunction with various monospecific antisera to detect urine components related to bladder cancer. All urine samples were centrifuged routinely, dialyzed and concentrated 10 times before assay by gel diffusion versus the various antisera. Urine was considered positive when it showed reactivity with 2 or more antibodies. This method of analysis resulted in the diagnosis of 64% of the bladder papillomas and 77% of the bladder cancers tested, compared to only a 7% falsely positive rate with normal urine. These data support the potential usefulness of an antiserum panel in the immunological diagnosis of bladder cancer in the general population and in high risk individuals.


Lymphocyte reactivity against virally transformed cells in patients with urologic cancer. Lymphocytes from patients with urologic cancer were tested in microcytotoxicity assays against human cells transformed by cytomegalovirus. Human lymphocytes were significantly cytotoxic against the transformed cell line when compared to a normal human control cell line. Patients with prostatic carcinoma demonstrated greater target cell reduction than those with benign prostatic hyperplasia.


Use of methylene blue in suspected colovesical fistula. Positive urine and blood assays of methylene and leukomethylene blue were obtained from 9 volunteers with normal colonic and renal function after a 100 ml, methylene blue enema (50 mg.). The study conclusively demonstrates that methylene blue is absorbed by rectal mucosa and excreted by the kidneys as a colored dye in the urine. Thus, intrarectal methylene blue cannot be used to confirm the diagnosis of a colovesical fistula.


Transurethral resection in a patient with a dysfibrinogen: fibrinogen cleveland I. A case of a congenitally abnormal fibrinogen that resulted in surgical wound dehiscence is reported. The patient underwent successful transurethral resection for benign prostatic hypertrophy while being treated with epsilon aminocaproic acid to prevent fibrinolysis of the abnormal fibrin clot.


Serological typing of Pseudomonas aeruginosa--comparison of various antigenic schema. Serotyping sera for both the agglutination (using IMSUT sera) and slide agglutination (using TIBS sera) were succesfully prepared using Homma's serotype strains. They were proved to be type specific and distinct. The two typing sera (IMSUT and TIBS) were used to determine the correspondence of major O-antigens among the different kinds of serotype schema which are used at present throughout the world. The serotype sera of Lanyi, Liu and Meitert were also used for the same purpose. The relationships among serotypes of the seven schemata were clarified as shown in tables in the text. Cross-reactions were found among several serotypes in each of several serotype schemata was suggested.


Serological typing of Pseudomonas aerugionosa--grouping of serotypes. Cross absorption and agar gel diffusion tests were performed using Homma and Liu standard strains and their antisera to study the antigenic relationships among the strains showing strong cross-agglutination. The results suggested that Homma 0:3 strain was able to produce a sufficiently high-titered antiserum against Liu 0:7 and 0:8, and that Liu 0:13 was identical to Homma 0:12. In the antisera against Liu 0:13 and 0:14 there existed a strong antibody common to both antigens as well as an antibody specific to each antigen. Furthermore, the presence of strong common antigens was demonstrated among Homma 0:2, 0:7, 0:13 and 0:16, and between Homma 0:15 and 0:17.


Oral non-absorbed antibiotics prevent infection in acute non-lymphoblastic leukaemia. 113 patients being treated for acute non-lymphoblastic leukaemia were investigated to determine the effect of suppression of body microbial flora on prevention of infection. They were randomly allocated to a control group or a group which received non-absorbed antibiotics by mouth and topical applications of cutaneous and mucosal antiseptic preparations. The group receiving oral non-absorbed antibiotics had significantly few infections, fewer deaths from infection, fewer pyrexial episodes, and consequently received less systemic antibiotic therapy than the controls.


Prolactin, oestrogen, and lipids in breast fluid. Concentrations of prolactin, oestrogens, and lipids were measured in the plasma and breast-duct fluid of 8 non-lactating premenopausal women without a known history of breast disease. Concentrations of prolactin, oestrogen, and triglycerides were significantly higher in ductal fluid than in plasma, but there was little difference with cholesterol concentration. The high concentrations of prolactin and oestrogen found in the ductal fluid of Western women may be relevant to development of breast cancer.


Growth-hormone deficiency in congenital rubella. Two boys with congenital rubella and concomitant growth-hormone deficiency responded to human-growth-hormone replacement and have already (over a period of 18 mo) achieved a significant increase in height. The association of these two diagnoses requires recognition so that treatable endocrine disorders, which are becoming recognized more often in congenital rubella, are always positively excluded when affected children with inexplicable short stature are seen.


Specific cell-mediated immune defect in active cytomegalovirus infection of young children and their mothers. 4 young children with active cytomegalovirus (C.M.V.) infection were  found, by an in-vitro lymphocyte-proliferation assay, to have a C.M.V.-specific cell-mediated immune defect. These children had antibodies to C.M.V. and were actively shedding C.M.V. in the urine when studied. Their general cellular immune responses were intact, with normal numbers of T lymphocytes and normal in-vitro responses to mitogens and at least one antigen. 3 of the 4 mothers studied shortly after delivery had decreased cell-mediated immunity to C.M.V. These findings suggest that an antigen-specific immune defect facilitates transmission of virus from mother to infant and permits persistence of viral replication in the offspring.


Respiratory allergy to urine proteins of rats and mice. Five atopic laboratory workers with asthma and other manifestations of allergy to rats and mice were sensitive to low-molecular-weight urine proteins--an alpha2-globulin in the rat and a prealbumin in the mouse. Positive skin prick and inhalation test reactions to the urine proteins were obtained; and radioallergosorbent tests (R.A.S.T.) for specific IgE antibody were also positive. These findings show that proper control of the collection and disposal of the animals' urine is necessary to minimise the likelihood of sensitisation.


Morphometric changes in rabbit ventricular myocardium produced by long-term beta-adrenoceptor blockade. In rabbits long-term beta-adrenoceptor blockade did not cause any change in the morphology of individual organelles in the ventricular myocardium. There were, however, striking and significant changes in their relative distribution: (1) the relative volume of the mitochondria was reduced and was replaced by an equivalent relative increase in sarcoplasm; and (2) the relative volume of vascular elements was increased and the interfibrillar volume was decreased, so that the path from capillary to cell wall was reduced. The relative volumes of myofibrils and nuclei were unchanged. The doses used were well within the range used clinically, so that the results could be relevant to the efficacy of beta-blockers in angina pectoris, because the relative increase in capillaries and shorter diffusion pathway for oxygen could raise oxygen availability. The reduced rate of growth of the heart in young rabbits during beta-blockade may also be relevant to the treatment of infants with hypertrophic obstructive cardiomyopathy by beta-blockers.