The search for "virogene" in human prostatic tissues: prostatic DNA polymerases. The "virogene-oncogene" hypothesis of Huebner and Todaro and the "provirus" hypothesis of Temin implicate RNA tumor viruses in the neoplastic transformation of mammalian cells. These hypotheses have been substantiated in several animal systems including primates and, presumably, in man. Because the detection in a tissue of one or two activities allegedly related to RNA tumor virus may not be conclusive evidence for viral presence, we have developed a scheme of coordinated morphologic, biologic, and biochemical investigations of human prostatic tissues. We report here the more recent progress we have made in one of the segments of our scheme of investigations. Two, possibly three, DNA polymerase activities from human prostatic tissue have been isolated and partially purified by DEAE-cellulose and phosphocellulose chromatography. These activities have been partially characterized. Based on template preferences and non-inhibition by selective inhibitors of reverse transcriptase, neither of the major polymerase activities appears to be the reverse transcriptase-type activity.


Properties of prostatic cultures transformed by SV40. SV-40-transformed hamster prostatic tissue has been previously evaluated as a model for human prostatic carcinoma. Because the original cell line was lost, Syrian golden hamster prostatic tissue has been established in explant culture and infected with a 10-6-cell tissue culture infectious dose (50 percent effective) of SV40. After in vitro transformation, the cells were produced in quantity and 60 times 10-6 cells were injected into adult male Syrian golden hamsters 24 hours after 400 rads of whole-body radiation. After 60-90 days, a small palpable tumor developed. These tumors could be serially transplanted in adult male animals without immunosuppression. The tumor cells were established in tissue culture and the cells were returned to adult animals without immunosuppression where they rapidly produced fast-growing tumors. The solid tumors were composed of sheets of pleomorphic polygonal cells with large nuclei and many nucleoli; they resembled undifferentiated human prostatic carcinoma. In vitro, the cultures contained small, rapidly growing cells with a population doubling time of about 1.3 days. The cells carried the SV 40-specific antigen. The modal chromosome number was 66-68 with a distribution of 47-120. Cells exposed to 2-bromo-5'-deoxyuridine in culture did not release particles with RNA-dependent DNA polymerase activity. Endocrine sensitivity in vivo and in vitro is undertermined to date.


Monolayer cultures of human prostatic cells. The objective of this study was to isolate, characterize, and preserve normal prostatic epithelial and carcinoma cell lines. To date, four specimens have been acquired from patients with benign prostatic hypertrophy. Culture attempts have been successful, yielding a total of 32 cell lines. Three of these lines have been examined thus far and were found to have clearly demonstrable tartrate-inhibitable acid phosphatase granules by histochemical techniques. Several problem areas emerge from these preliminary studies. These involve the difficulty of (a) characterization of the cells as originating from prostatic epithelium, (b) identification of prostatic carcinoma cells, and (c) possible senescence in culture of cell lines.


A study of alpha1-fetoprotein levels during exposure to 3'-methyl-4-dimethylaminoazobenzene and its analogs. The serum concentration of alpha1-fetoprotein (alpha1F) was determined in rats following exposure to the hepatocarcinogen, 3'-methyl-4-dimethylaminoazobenzene, and its analogs. Small quantities of the carcinogen caused a rapid and significant elevation of alpha1F. Neither 2-methyl-4-dimethylaminoazobenzene nor p-aminoazobenzene resulted in any elevation of alpha1F. Further, under circumstances wherein 2-methyl-4-dimeth-laminoazobenzene is reported to become carcinogenic, i.e., when administered at the time of 70 percent hepatectomy, neither elevation of alpha1F nor histological alteration of the liver was noted. The increase in alpha1F after 3'-methyl-4-dimethylaminoazobenzene exposure is the result of a highly selective interaction. The possible contribution of hepatocyte mitosis to the elecation of alpha1F seen during chemical carcinogenesis is emphasized.


Effect of 3-methylcholanthrene (NSC-21970) on human prostate in organ culture. Long-term organ culture of normal human prostatic tissue with demonstration of functional and morphologic maintenance of the glandular epithelium is described. The carcinogen 3-methylcholanthrene at a level of 4 mug/ml in the culture fluid induced morphologic changes that were not observed in untreated cultures. The changes observed after exposure to the carcinogen for 15-30 days included hyperplasia and anaplasia of the glandular epithelial cells with large variation in nuclear size and shape. These changes were suggestive of a neoplastic potential.


Induction of the polyamine-biosynthetic enzymes in mouse epidermis by tumor-promoting agents. A single topical application of 1.0 mg of crotol oil or 17 nmoles of 12-O-tetradecanoyl-phorbol-13-acetate (TPA) resulted in a rapid, transient stimulation of mouse epidermal ornithine decarboxylase activity. The activity reached a peak (230-fold greater than control after TPA) at 4 to 5 hr after croton oil or TPA treatment and returned to control level by 12 hr. The stimulation of S-adenosyl-L-methionine decarboxylase activity was less pronounced, reaching a peak of activity (6- to 7-fold greater than control) at 9 to 12 hr after TPA or croton oil and slowly declining to control level. The stimulation of both enzyme activities was dependent on the dose of TPA applied and correlated well with the promoting ability of these doses on mouse skin. Phorbol, the nonpromoting parent alcohol of TPA, did not affect the enzymes activities. Cycloheximide pretreatment abolished the increase in enzyme activities after TPA application. By measuring the decline of enzyme activity following cycloheximide treatment, enzyme half-lives of 17 and 41 min were obtained for ornithine and S-adenosyl-L-methionine decarboxylase, respectively. 5-Azacytidine pretreatment prevented the stimulation of enzyme activities by TPA, while actinomycin D had no effect. Cordycepin (3'-deoxyadenosine) partially blocked the rise in enzyme activities.


Tumor antigen and acid phosphatase isoenzyme in prostatic cancer. Plasma and prostatic fluid from man, dog, and baboon were measured for carcinoembryonic antigen (CEA) by a radioimmunoassay technique. No CEA was detected in plasma, prostatic fluid, or seminal fluid in 12 dogs and three baboons. Elevated CEA (less than 2.5 ng/ml) was found in 13 of 20 human prostatic fluids. It was inferred that there was no immunologic cross-reactivity of CEA among man, dog, and baboon. CEA has been isolated and purified from liver tumors. Biochemical studies reveal that CEA consists of 60 percent carbohydrate and 40 percent protein. It contains the following carbohydrates: fucose, mannose, galactose, sialic acid, N-acetylglucosamine, and a small amount of N-acetylgalactosamine. The following amino acids were found in CEA: lysine, histidine, arginine, aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, emthionine, isoleucine, leucine, tyrosine, phenylalanine, and cysteine. The amino acid sequence (first 30 amino acids) of the N-terminal has been determined. The N-terminal amino acid was lysine. Using this study as a model, other tumor antigens from prostatic tumor tissues are being investigated. The acid phosphatase isoenzyme from prostatic tissue was also studied. After a series of purifications, two chromatographic fractions were obtained. Treatment with neuraminidase removed the sialic acid content of the molecule, changed the isoelectric focusing patterns, and abolished the chromatographic heterogeneity. Sedimentation studies indicated a molecular weight of about 100,000. Biochemical studies showed that prostatic acid phosphatase isoenzyme is a glycoprotein which consists of 7 percent carbohydrate and 93 percent protein. It contains fucose, galactose, mannose, sialic acid, N-acetylglucosamine, and the following amino acids: aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine, tryptophan, and cysteine. An antiserum to this purified prostatic acid phosphatase isoenzyme is being prepared in animals.


The binding of carcinoembryonic antigen by antibody and its fragments. In order to assess the potency of antigenic fragments of carcinoembryonic antigen (CEA) in the radioimmune assay; it is necessary to know whether the high affinity of goat anti-CEA antibody (which makes possible the detection of as little as 10--11M CEA) is due to bivalent binding of the CEA molecule. Immunoglobulin G and the F(ab')2 and Fab fragments derived from it were prepared from an anti-CEA serum and tested for their abioity to bind CEA. Equivalent concentrations of binding sites of the bivalent F(ab)2 and univalent Fab fragments of anti-CEA were identical to the immunoglobulin G fraction in the standard inhibition curve. Fragments of CEA obtained by trypsin digestion produced equivalent inhigition curves when tested with either immunoglobulin G, F(ab')2, or Fab". This, increased avidity due to bivalent binding to a single antigen molecule cannot be invoked to explain the sensitivity observed in the CEA assay. This high sensitivity implicates the protein rather than the carbohydrate as an important part of the antigenic determinant(s) of CEA.


An ultrastructural investigation of the spermatozoa of two ophiuroids, Ophiocoma echinata and Ophiocoma wendti: acrosomal morphology and reaction. The spermatozoa of two closely related species of ophiuroids, Ophiocoma echinata and Ophiocoma wendti, were examined ultrastructurally. Morphologically, these spermatozoa resemble those of other non-echinoid echinoderms. The acrosomal complex, completely contained within an anterior fossa in the spherical nucleus, consists of a membrane-limited acrosomal vesicle and periacrosomal material. Events of the acrosomal reaction in O. echinata and O. wendti are presented. In both species, the reaction results in the establishment of an extracellular coat of acrosomal vesicle origin on the anterior surface of the spermatozoon. The possible role of this extracellular coat in the species-specific binding of sperm and ova is discusses. The origin of acrosomal tubule membrane is elucidated.


A quantitative electron microscopic analysis of the keratinizing epithelium of noral human hard palate. The epithelium of normal human hard palate was subjected to sterologic analysis. Ten biosies were selected from a total of twenty specimens collected from 9 to 16 year old females, and processed for light- and electron microscopy. At two levels of magnification, electron micrographs were sampled from three strata (basale, spinosum, granulosum) in two locations (epithelial ridges and portions over connective tissue papillae). Stereologic point counting procedures were employed to analyse a total 1560 electron micrographs. In general, the thickness of the palate epithelium was 0.12 mm (over papillae) and 0.31 mm (in ridges), the epithelium is distinctly stratified, and homogeneously ortho-keratinized. From basal to granular layers, the composition of strata revealed decreasing densities of nuclei, mitochondria, membrane-bound organelles and aggregates of free ribosomes. Keratohyalin bodies and membrane coating granules increased, and cytoplasmic filaments with a constant diameter of about 85 A increased from 14 to 30% of cytoplasmic unit volume. The cytoplasmic ground substance occupied a stable 50% of the epithelial cytoplasm in all strata. The composition of basal layers in ridges differed from that over connective tissue papillae. The data are discussed in relation to the observations that (1) an increasing gradient of filament density is not the most characteristic feature of ortho-keratinizing oral epithelium and (2) differences in the degree of differentiation in cells of the stratum basale coincided with the comparable frequency distribution pattern of dividing cells.


Ultrastructure of cell surface coat (glycocalyx) in rat urinary bladder epithelium. Earlier statements to the contrary, the present study demonstrates the presence of a cell surface coat (glycocalyx) on the luminal plasma membrane of the superficial transitional epithelial cells lining the urinary bladder of male Buffalo rats. This coat was demonstrated with ruthenium red, an electron dense stain, which revealed a surface layer, 60-80 A thick, separated from the outer leaflet of the plasma membrane by an electron lucent layer, approximately 30 A thick. The structure of the glycocalyx was not affected by 12 weeks of treatment with dibutylnitrosamine, a known bladder carcinogen.


The location of the genes coding for hexon and fiber proteins in adenovirus DNA. A serological analysis has been made of the capsid antigens hexon and fiber from 17 Ad5-Ad2+ND1 recombinants that enables us to determine the phenotype of the recombinants. By correlation of this data with the genetic and physical maps of the adenovirus genome, obtained by recombination and restriction endonuclease analysis, the genes coding for the hexon and fiber have been assigned to specific locations on the adenovirus DNA.


Malignant hypertension due to musculo-mucoid intimal hyperplasia of intrarenal arteries. Absence of renal fibrinoid necrosis. Severe essential hypertension in a subset of American black subjects is associated with marked stenosis of interlobular arteries and arterioles of the kidneys, observed by renal biopsy and binephrectomy specimens. The interlobular arterial stenosis is caused by marked thickening of the intima due mainly to the presence of smooth muscle cells, basement membrane material, and acid mucopolysaccharide. Because of this makeup, we propose the term "musculo-mucoid intimal hyperplasia" for this lesion. The media of these arteries appears maximally dilated, and by electron microscope displays degenerative changes of the smooth muscle cells. The arterioles are thickened, due mainly to hyalinization, but also due to the musculo-mucoid change (onionskin effect). The smooth muscle cells are degenerated and atrophic. These patients do not exhibit fibrinoid necrosis of the arteries, arterioles, and glomeruli, presumably because of the rapidity of the development of the arterial stenotic lesion. Accordingly, the glomeruli are destroyed by ischemia, and there is no evidence of glomerulitis (no "Kombinations" form of Fahr). A unifying hypothesis concerning renal hypertensive arterial disease is suggested by these studies. This hypothesis places the main emphasis for all the morphological expressions of the intrinsic visceral vasculature on changes involving the main functional unit of the vessel wall, the medical smooth muscle.


Observations on blood viscosity changes after acute myocardial infarction. Serial blood rheologic measurements were made in 25 patients with acute myocardial infarction; measurements included blood and plasma viscosities, hematological data and plasma protein concentrations. The blood viscosity was elevated on admission and for more than 21 days after acute myocardial infarction. However, the cause of the elevated viscosity was changed as a function of time after acute myocardial infarction. During the first three days after admission, the high blood viscosity was mainly attributable to high hematocrit values. Thereafter, the hematocrit fell, but blood viscosity remained high. High blood viscosity after the first three days of acute myocardial infarction can be correlated with increases in plasma viscosity and red cell aggregation, which in turn are explained by elevations of alpha 2 globulin and fibrinogen concentrations. Patients with higher blood viscosity on admission had a significantly higher incidence of complications, i.e., shock, thromboembolism and left ventricular failure.


Echocardiography of the intra-atrial baffle in dextro-transposition of the great vessels. Twelve patients with dextro-transposition of the great vessels (age eight months to four years) were studied by echocardiography following Mustard's procedure. Nine of them had also been studied preoperatively. Postoperatively all patients demonstrated structural echoes in the atrial cavity behind the pulmonary root. In ten, the motion pattern generally resembled that of a stenotic atrioventricular valve iwth a sharp anterior movement followed by flattening in diastole and rapid posterior excursion in systole. The maximum amplitude of motion ranged from 4 to 9 mm (average 6.6 mm). In the remaining two cases, the anterior diastolic movement was attenuated. Similar moving, linear echoes with larger amplitudes of motion (10-14 mm) were observed behind the tricuspid valve in four patients while poorly moving, multiple or thick conglomerate echoes (2-11 mm wide) were detected in seven cases. Echocardiographic contrast studies performed by injecting indocyanine green via catheters placed on either side of the intra-atrial baffle identified it as the source of these echoes. Following operation, coarse diastolic undulations of the mitral valve (ten cases) and the tricuspid valve (nine cases) were noted. Also, fine flutter of both atrioventricular valves, not present before, appeared after operation in three patients. These findings may be related to the altered pathway of blood flow and turbulence resulting from the insertion of the baffle in the atria. Echocardiography appears useful in delineating the character and movement pattern of the intra-atrial baffle and this may have potential in evaluating its long-term functional status.


Anomalous left coronary artery originating from the pulmonary artery. Report on 15 cases. Fifteen infants and children with the diagnosis of anomalous left coronary artery from the pulmonary trunk have been encountered at the Children's Hospital Medical Center, Boston, Massachusetts from 1958 to 1973. After thorough clinical and laboratory evaluation, they have been treated by anticongestive measures. Nine patients have had ligation of the anomalous left coronary artery at its entrance into the pulmonary artery; one patient has undergone coronary bypass surgery. The lelctrocardiogram proved to be the most helpful diagnostic clinical laboratory test, Vectorcardiograms are valuable not only in diagnosis but also in the follow-up of the patients from the prognostic point of view. The most sensitive tool for the definitive diagnosis is an aortic rool angiogram; we have no false negatives or false positives with this method. The twelve patients with complete cardiac catheterization data could be divided into three groups, according to the pressure and magnitude of the left-to-right shunt at the pulmonary level. All patients with an appreciable le?T-TO-RIGHT SHUNT SURVIVED. Patients in whom no left-to-right shunt could be demonstrated by angiography died. Half of the patients with only small left-to-right shunt survived; The results of surgical and medical treatment, were identical within the three groups. Medical management in infancy, according to coronary care principles, with definitive surgical correction at a later age, is the preferred treatment. Ligation of the anomalous left coronary artery is recommended in severely symptomatic infants with documented left-to-right shunt at the pulmonary artery level, who do not respond to medical management.


Cardiac involvement in Whipple's disease. Although cardiac involvement in Whipple's disease has been reported previously, the extent and significance of such involvement has not been stressed. In our series of 19 patients, 58% had clinical cardiac findings and 79% had gross cardiac lesions at autopsy. Histologically there were PAS-positive macrophages in the pericardium, myocardium, and valves of each of these patients. These collections of macrophages were associated with chronic inflammatory cells and foci of fibrosis with resultant adhesive pericarditis, focal myocardial fibrosis similar to the fibrosis of idiopathic cardiomyopathy, and valvular fibrosis with deformity grossly resembling the valvular lesions of chronic rheumatic heart disease. Four of the patients with mitral valvular deformity had cardiac murmurs, and ECG changes were noted in six patients with mitral myocardial fibrosis. In addition, pericardial friction rubs were heard in two patients. By electron microscopy rod-shaped bodies, presumably bacteria, were noted in the mitral valve and myocardium. The structures are identical to those described in the small intestinal mucosa of patients with Whipple's disease and presumably are the causative agent of the pancardiopathy.


Relative usefulness of measuring polyamines in serum, plasma, and urine as biochemical markers of cancer. Serial samples of plasma and serum were collected in the morning and afternoon from cancer patients, along with 24-h urine specimens. Values for serum and plasma samples taken at the same time from the same patient differed little, suggesting that either procedure is acceptable for polyamine analysis. Increases in concentrations of putrescine and spermidine in serum and plasma correlate well with such increases in 24-h urine samples. Spermidine concentrations in sera were consistently about 10-fold lower than corresponding urine values. Putrescine concentrations were 10- to 100-fold different. The variation in putrescine values may be due to its more active metabolism by diamine oxidase, an enzyme known to be present in serum. Concentrations of polyamines in serum and urine increased in response to effective chemotherapy.


Serum triiodothyronine 7-15 years after fractionated low dose radioiodine therapy of thyrotoxicosis. In 189 of 334 patients, who had been treated with fractionated doses of radioiodine for Graves' disease 7-15 years ago, the serum concentrations of triiodothyronine have been estimated in additition to the following parameters: Protein bound 127-iodine (PB-127-I), free thyroxine index, cholesterol- and TSH-level in serum, tendon reflex time and clinical index according to Billewitz et al. (1969). In forty-one of the 189 sera the free T4 (AFT4) and free T3 (AFT3) concentrations were measured as well. The following hormonal and clinical patterns were observed: (1) Euthyroidism, with all parameters within the normal range in 148 patients (equals 78-4%). (2) Hypothyroidism with low serum T3, PBI, AFT4, AFT3 and elevated TSH six subjects (equals 3-2%). (3) Persistent hyperthyroidism with increased thyroid hormone concentrations and low TSH in seven cases (equals 3-7%). (4) In twenty-eight clinically euthyroid patients (equals 14-8%) TSH was elevated with normal PBI and AFT4. Twenty of these subjects had a low normal T3 and in nine the T3 was clearly in the hypothyroid range (40 plus or minus 12 mug/dl). (5) The constellation of normal T3, low T4 and elevated TSH, which has been frequently found after radioiodine therapy, has been seen in only moderate form in ten cases.


Amniotic fluid alphafetoprotein measurements in the early prenatal diagnosis of central nervous system disorders. Amniotic fluid alphafetoprotein (AFP) has been measured in 520 pregnancies between 8 and 24 weeks of gestation. The normal range of values has been defined for fortnightly periods between these limits. Grossly elevated AFP concentrations were found in four pregnancies leading to spina bifida and nine pregnancies leading to anencephaly. Slightly elevated AFP concentrations were found in one twin pregnancy and two pregnancies affected by rhesus isoimmunisation. Normal AFP values were observed in 36 amniotic fluids from pregnancies where the outcome was rhesus isoimmunisation, an inborn error of metabolism, a cytogenetic disorder or a birth defect unrelated to the central nervous system. The reliability of amniotic fluid AFP in the early prenatal diagnosis of spina bifida and anencephaly and the possibility of performing assays on samples sent by post from any part of the world are emphasized.


Alpha fetoprotein levels in maternal serum and in amniotic fluid from early normal pregnancies. Alpha fetoprotein (AFP) was determined in serum from 241 women in the 8th to 43rd week of gestation by quantitative radioimmunoelectrophoresis, and in 103 amniotic fluid samples from the first half of pregnancy by rocket immunoelectrophoresis. Both serum and amniotic fluid samples were obtained from a larger material, but samples were included only from pregnant women whose infants were normal at birth. The 90% normal AFP range was calculated both for maternal serum and for amniotic fluid.


Elevated levels of alfa fetoprotein in maternal serum and amniotic fluid in two cases of spina bifida. Therapeutic abortions were performed in two cases of spina bifida which were diagnosed by determination of the lafa fetoprotein levels in the amniotic fluid and maternal serum.


Synovial fluid analysis.

Synovial fluid analysis. Synovial fluid analysis plays an important role in the differential diagnosis of various forms of arthritis. The results of macroscopic, biochemical, bacteriological, and microscopic techniques of evaluating synovial fluid must be correlated to establish the diagnosis of each of the various forms of arthritis. The principle of polarized microscopy as applied to the differential diagnosis of crystalline arthritis is currently of special interest.


The effect of short-term immobilization on the rabbit knee joint cartilage. A histochemical study. Rabbit knee joints were immobilized in the extended position for 1 to 6 days, and examined by means of histochemical techniques. In the articular cartilage matrix, the earliest change was the loss of affinity for metachromatic stains, confirming earlier observations. The affinity of cartilage for high iron diamine and alcian blue stains was also diminished, although less dramatically; these stains demonstrated the concentration of glycosaminoglycans more accurately than metachromatic stains. With the colloidal iron method, changes were seen in the chondrocytes before there was any loss of affinity for metachromatic stains. Following only 1 or 2 days of immobilization, the chondrocytes in the gliding surface of the joint stained intensely with colloidal iron whereas those in the controls stained very minimally.


Morphological and biochemical studies during differentiation and calcification of fracture callus cartilage. Differentiation and calcification of cartilage of a fracture callus morphologically, ultrastructurally, and histochemically resembles cartilage of growing epiphyseal plate. The fracture callus includes the various cartilage cell types found in the epiphyseal plate. Proliferating and hypertrophic cartilage had higher activities of cytochrome oxidase, alkaline phosphatase and glutamate aspartate transaminase than fibrocartilage. Enzymes controlling glycogen synthesis and glycolysis had higher levels of activity in fibrocartilage than in hypertrophic cartilage. Lysosomal enzymes, catalase, 6-phospho-gluconic acid and glucose 6-phosphate dehydrogenase were uniformly distributed. Alkaline phosphatase was associated with extracellular vesicles found in hypertrophic cartilage. EM dense granules were found in mitochondria in hypertrophic cartilage. There was an increase of total lipids in hypertropic and calcified cartilage as compared to resting cartilage.


Dihydrotachysterol-induced aortic calcification. A histochemical and ultrastructural investigation. Early dihydrotachysterol-induced calcification of the rat aorta occurs in elastic lamellae. The first deposition of inorganic substance leads to the formation of very thin filament-like structures of low electron density. The characteristic shape of these structures suggests that they could correspond to calcified filamentous components of the elastic tissue. When calcification spreads from the calcified elastic lamellae into the adjacent tissue, the inorganic substance is initially collected in roundish structures, probably of cellular origin, and is successively laid down in the entire matrix of the aortic wall, including collagen fibrils. All the calcified areas contain glycoproteins and acid proteoglycans. A cartilage-like tissue often develops near calcified areas. Its fine structure is very similar to that of the normal hyaline cartilage. It can be calcified, but usually the inorganic substance is not crystalline as it is in normal cartilage. It seems to consist of very small, linearly aggregated inorganic granules which form irregular structures. These seem to develop in close relationship with the fibrillar, probably collagenic, network of the matrix. No ultrastructural findings have been obtained for explaining cartilage induction near calcified areas of the aortic wall. It is possible that cartilage differentiation is regulated by diffusible substances which cannot be recognized under the electron microscope.


Identical giemsa banding patterns of two macaca species: Macaca mulatta and M. fascicularis. A densitometric study. Karyotypes of Macaca mulatta and M. fascicularis (2n equals 42) were compared using the Giemsa banding pattern technique. No differences between the chromosomes of both Macaca species were found with conventional staining methods. In the literature the occurrence of fertile hybrids is used as an argument in favor of conspecificity. The banding patterns were evaluated by comparison of their density distribution curves. Our data indicated no significant differences between the two Macaca species.


Hoechst 33258 fluorescent staining of Drosophila chromosomes. Metaphase chromosomes of D. melanogaster, D. virilis and D. eopydei were sequentilly stained with quinacrine, 33258 Hoechst and Giemsa and photographed after each step. Hoechst stained chromosomes fluoresced much brighter and with different banding patterns than quinacrine stained ones. In contrast to mammalian chromosomes, Drosophia's quinacrine and Hoechst bright bands are all in centric heterochromatin and the banding patterns seem more taxonomically divergent than external morphological characteristics. Hoechst stained D. melanogaster chromosomes show unprecedented longitudinal differentiation by the heterochromatic regions; each arm of each autosome can be unambiguously identified and the Y shows eleven bright bands. The Hoechst stained Y can also be identified in polytene chromocenters. Centric alpha heterochromatin of each D. virilis autosome is composed of two blocks which can be differtiated by a combination of quinacrine and Hoechst staining. The distal block is always Q-H- while the proximal block is, for the various autosomes, either Q-H-, Q+H- or Q+H+. With these permutations of Hoechst and quinacrine staining, D. virilis autosomes can be unambiguously distinguished. The X and two autosomes have H+ heterochromatin which can easily be seen in polytene and interphase nuclei where it seems to aggregate and exclude H- heterochromatin. This affinity of fluorochrome similar heterochromatin was been seen in colcemide induced multiple somatic non-disjunctions where H+ chromosomes were distributed to one rosette and H- chromosomes were distributed to another. Knowing the base composition and base sequences of Drosophila satellites, we conclude that AT richness may be necessary but is certainly an insufficient requirement for quinacrine bright chromatin while GC richness may be a sufficient requirement for the absence of quinacrine or Hoechst brightness. Condensed euchromatin is almost as bright as Q+ heterochromatin. While chromatin condensation has little effect on Hoechst staining, it appears to be "the most important factor responsible for quinacrine brightness.' All existing data from D. virilis indicate that each fluorochrome distinct block of alpha heterochromatin may contain a single a single DNA molecule which is one heptanucleotide repeated two million times.


Location of nucleolar organizers in animal and plant chromosomes by means of an improved N-banding technique. With an improved N-banding technique, the location of nucleolar organizing region was determined in 27 kinds of material including mammals, a marsupial, birds, amphibians, fishes, an insect and plants. In most cases the N-bands were clearly located on certain specific regions of chromosomes, such as the secondary constriction, satellite, centromere, telomere and heterochromatic segment, while in some species they were detected as very minute bodies distributed over many chromosomes. From the available cytological and biochemical data it was suggested that the N-bands represent certain structural non-histone proteins specifically linked to nucleolar organizers in various eukaryotic chromosomes.


Homologous bands on the long arms of the X and Y chromosomes of Anopheles atroparvus. The long arms of the X and Y chromosomes of the mosquito Anopheles atroparvus (2n equals 6) are equal in length, synchronous in their late DNA replication and have homologous G AND Q bands. This indicates that differentiation of the two sex chromosomes was the consequence of a single deletion of an autosome to give the Y chromosome, not followed by the acquisition of differential heterochromatic blocks.


Comparison of G-, Q- and EM-banding patterns exhibited by the chromosome complement of the Indian muntjac, Muntiacus muntjak, with reference to nuclear DNA content and chromatin ultrastructure. When the chromosomes of the Indian muntjac, Muntiacus muntjak, were compared following treatment with two presently used banding methods, trypsin-Giemsa (G) and quinacrine-hydrochloride (Q) with structural bands as seen in the electron microscope, definite correlations were observed with respect to the numbers and positions of individual bands. - Weights obtained for the individual chromosomes were: No. 1, 9.98 pg; No. 2, 4.10 pg; No. 3, 4.43 pg; No. 3-X, 5.05 pg; and Y, 0.55 pg. Average diameters and weights for individual fibers were 193 A and 8.74 times 10-16 g/micron, respectively, for stimulated metaphase chromosomes and 185 A and 8.73 times 10-16 g/micron, respectively, for unstimulated chromosomes. Fibers of interphase nuclei exhibited an average diameter of 191 A and a weight of 5.87 times 10-16 g/micron. - The total amound of nuclear DNA present in interphase nuclei was 3.88 pg.


Chromosome banding pattern conservatism in birds and nonhomology of chromosome banding patterns between birds, turtles, snakes and amphibians. The G-banded karyotypes of 4 species of birds representing the orders Galliformes, Columbiformes and Musophagiformes were compared. Banding pattern homology between orders was limited t 5o 5 major chromosome arms and the Z chromosome. Even in these major chromosome arms pericentric and paracentric inversions produced alteration of the banding pattern sequences. Addition of constitutive heterochromatin was responsible for changes in banding patterns in the Z chromosome. The chromosome banding patterns of an emydid turtle, Terrepene carolina, 5 species of boid snakes of the genera Liasis, Acrantophis, and Sanzinia and the African clawed-frog. Xenopus muelleri, were also compared to the bird chromosome banding patterns. No homology was observed between any of these major groups: bird, snake, turtle, amphibian. However, intergroup homology was apparent. - The data obtained do not support reports of broad interordinal direct homology of the macrochromosomes of birds and refutes the idea of a primitive bird karyotype with 3 pairs of "Agroup' chromosomes and 3 pairs of "B group' chromosomes. - The major mechanisms responsible for chromosome evolution in birds appear to be centric and tandem fusions, paracentric and pericentric inversions, and addition or deletion of heterochromatin.


Mechanisms of chromosome banding. IV. Optical properties of the Giemsa dyes. A thorough understanding of the mechanisms of R-, C- and G-banding will come only from studies of the binding of Giemsa dyes to isolated and characterized preparations of heterochromatin and euchromatin. Since such studies require an exact knowledge of the optical characteristics of Giemsa, the spectral absorption curves and extinction coefficients of Giemsa and its component dyes at various concentrations in the presence and absence of DNA were determined. - Although Giemsa is a complex mixture of thiazin dyes plus eosin; methylene blue, and azure A, B or C alone gave good banding. Thionin, with no methyl groups, gave poor or no banding. Eosin was not necessary component for banding. - The most striking characteristic of the thiazin dyes is that they are strongly metachromatic, i.e., their absorption spectra and extinction coefficients change as the concentration of the dye increases or as they bind to positively charged compounds (chromotropes). These changes, especially for methylene blue, are described in detail and allow a distinction between concentration dependent binding to DNA by intercalation and binding by side stacking.


Mechanisms of chromosome banding. V. Quinacrine banding. A series of biochemical investigations were undertaken to determine the mechanism of Q-banding. The results were as follows: 1. In agreement with previous studies, highly AT-rich DNA, such as poly(dA)-poly(dT), markedly enhanced quinacrine fluorescence while GC containing DNA quenched fluorescence. These effects persisted at DNA concentrations comparable to those in the metaphase chromosome. 2. Studies of quinacrine-DNA complexes in regard to the hypochromism of quanacrine, DNA Tm, DNA viscosity, and equilibrium dialysis, indicated the quinacrine was bound be intercalation with relatively little sid binding. 3. Single or double stranded nucleotide polymers, in the form of complete or partial helices, were 1000-fold more effective in quenching than solutions of single nucleotides, suggesting that base stacking is required for quenching. 4. Studies of polymers in the A conformation, such as transfer RNA and DNA-RNA hybrids, indicated that marked base tilting does not affect the ability of nuclei acids to cause quenching or enhancement of quinacrine fluorescence. 5. Salts inhibit the binding of quinacrine to DNA. 6. Spermine, polylysine and polyarginine, which bind in the small groove of DNA, inhibited quinacrine binding and quenching, while histones, which probably bind in the large groove, had little effect. This correlated with the observation that removal of histones with acid has no effect on Q-banding. 7. Mouse liver chromatin was separated into five fractions. At concentrations of quinacrine from 2 times 10-6 to 2 times 10-5 M all fractions inhibited to varying degrees the ability of the chromatin DNA to bind quinacrine and quench quinacrine fluorescence. At saturating levels of quinacrine two fractions, the 400 g pellet (rich in heterochromatin) and a dispersed  euchromatin supernatant fraction, showed a decreased number of binding sites for quinacrine. These two fractions were also the richest in non-histone proteins. 8. DNA  isolated from the different fractions all showed identical quenching of quinacrine fluorescenc. 9. Mouse GC-rich, mid-band, AT-rich, and satellite DNA, isolated by CsCL AND Cs-2SO-4-Ag+ centrifugation all showed identical quenching of quinacrine fluorescence, indicating that within a given organism, except for very AT or GC-rich satellites, the variation in base composition is not adequate to explain Q-banding. We interpret these results to indicate that: (a) quinacrine binds to chromatin by intercalation of the three planar rings with the large group at position 9 lying in the small groove of DNA, (b) most pale staining regions are due to a decrease binding of quinacrine, and (c) this inhibition of binding is predominately due to non-histone proteins.


[Recurrent ventricular fibrillation (author's transl)]. 106 episodes of ventricular fibrillation were observed in 11 patients, 9 of whom had acute myocardial infarctions. The heart rate before the onset of ventricular fibrillation was below 50 in two episodes, 60 to 100 in 63, and above 100 in 41. 83 attacks of ventricular fibrillation were preceded by fewer than five ventricular premature beats, 23 by more than five, 19 by more than ten. Multifocal ventricular premature beats occurred ten times, runs of ventricular premature beats 11 times, with only three falling into the vulnerable period. Left bundle branch block was present in three patients, right bundle branch block with left anterior hemiblock in two, isolated left anterior hemiblock in two, left posterior hemiblock in one. 85 episodes of ventricular fibrillation occurred during antiarrhythmic treatment, 72 during lidocaine administration. Antiarrhythmic drugs were effective only in reducing the number of ventricular premature beats. The only successful treatment of recurrent episodes of ventricular fibrillation was repeated electrical countershocks.


[New chemotherapy (adriamycin, belomycin and vincristin) of metastasizing malignant testicular teratoma (author's transl)]. In a prospective study 18 patients with metastasizing testicular teratomas were treated with adriamycin, bleomycin and vincristin. Sixteen could be fully evaluated: 12 in stage IV (visceral metastases), three in stage III (lymphondular involvement beyond the diaphragm) and one with advanced stage II (iliac, para-aortic and inguinal nodes). None of the patients had previously received chemotherapy. Toxic side-effects were not severe enough to prevent 12 patients from being treated on an outpatient basis. Complete remission occurred in five, with four of them still in remission two, eight, ten and eleven months later. Partial remission (50% decrease in tumour size for at least four weeks) occurred in three, with one still improving. No change was noted for 11 months in a patient with advanced pulmonary disease. Progression under therapy occurred in seven patients and in some of them there was an initial response which did not, however, meet the criteria of partial remission. The therapeutic results as well as toxicity suggest that this regime is superior to actinomycin D montherapy and various previously used combinations.


Tracing neuronal connections with radioisotopes applied extracellularly. Methodological and technical problems of the autoradiographic-neuroanatomical tracing (ARNT) technique are discussed. The size of the labeled area after a tritiated amino acid injection varies directly with the volume of isotope, the rate of injection, and the length of exposure of the secretion to the emulsion. Frozen sections can be used for autoradiography if they are mounted on subbed slides, dehydrated in ethanol, defatted for 1 hour in xylene, rehydrated through ethanol and water, and dried before coating with emulsion. Brushes used for mounting frozen sections should be used only for this purpose and dipped in boiling distilled water before use to avoid chemoreduced streaks in the emulsion due to contamination from the brushes. Excessive dilution of Kodak emulsion can leave less than a monolayer of grains over certain types of sections; the emulsion thickness should be checked by exposing coated test sections to a brief flash of light and developing immediately. A high intensity safelight recommended for the ARNT darkroom is the Thomas Duplex Super monochromatic sodium vapor bulb safelight; for Kodak NTB-2, -3 and Ilford L4 emulsions the red-banded and yellow-banded filters are used. A useful stain combination for ARNT is Luxol fast blue stained before coating and cresyl violet stained after developing which demonstrates both neuronal cell bodies and myelinated tracts in the same section.


The retrograde transport method.

The retrograde transport method. After an injection of horseradish peroxidase into the brain, the enzyme product can later be found in cell bodies of neurons which project axons to the injection site. The use of the tracer has now been adapted as a means of identifying neurons on the basis of their efferent fiber projections. In this review particular attention is paid to characteristics of the phenomenon of retrograde transport, such as diffusion of the marker from the injection site and the region of effective uptake, uptake by injured axons of passage as well as by axon terminals and the preterminal portions of axons, and the anterograde movement of peroxidase. The retrograde transport method is compared to the previously available method of retrograde cell degeneration and the advantages and limitations of both techniques are considered.


Detection and localization of specific antigens in the reproductive tracts of cycling, pregnant, and ovariectomized hamsters. A systematic search was made for components specific to the female reproductive tract in golden hamsters. Antisera produced in rabbits against saline homogenates of hamster uteri (collected on the night of estrus) cross-reacted extensively with extracts of 12 other tissues in agar gel double-diffusion assays. Absorption of the antisera with small intestine, lung, and liver rendered the immune sera specific for uterine and oviductal antigens (within the limits of the sensitivity of the precipitin assays). Immunoelectrophoretic analysis resolved 12 uterine antigens, many of which were similar to components in several other tissues. Absorbed antisera specific for reproductive tract antigens formed one postalbumin arc with uterine and oviductal extracts in immunoelectrophoretic studies. No reactions were detected between specific antisera and five other organ extracts or plasma. An indirect immunofluorescent antibody technique was used to detect changes in the distribution of specific antigens in reproductive tracts of cycling, pregnant, and ovariectomized hamsters. The gamma-globulin fraction of anti-uterus sera (absorbed with small intestine, lung, and liver), shown to be specific for reproductive tract tissues in precipitin tests, was used to localize antigens. Appropriate controls indicated that the fluorescence observed was due to antigen-antibody interactions. During the cycle, specific antigens were usually confined to the ampullary lamina propria, except during estrus, when they were prominent in the lamina propria and luminal epithelium of the ampula. Specific antigens were never abundant in the isthmus of nonpregnant hamsters. On day 1 postcoitum, the components were found throughout the ampullary and isthmic regions. By day 2 postcoitum, ampullary antigens were usually confined to the lamina propria. The specific components were not prominent in the oviduct on day 3 postcoitum, but were conspicuous in both ampulla and isthmus on day 4. Specific antigens in the uterus were confined to endometrial glands in nonpregnant animals during proestrus, estrus, and (occasionally) metestrus. Diestrous uteri contained no specific antigens. During the first 2 days of pregnancy, antigens were not abundant and were usually confined to the glands and stroma. On days 3 and 4 of pregnancy the specific antigens were prominent in the endometrial glands and stroma and along the apical borders of some luminal epithelial cells. By day 5, these components were less conspicuous in all areas of the endometrium. Uteri of spayed animals receiving no hormones or estradiol alone lacked the specific antigens. However, progesterone (after estrogen priming) promoted the appearance of these components, and the distribution resembled that seen in uteri of 3- and 4-day pregnant animals.


Progestogen-induced diabetes in the dog. The development of diabetes in a small percentage of female beagles receiving large doses of synthetic progestogen for one year is described. The abnormalities in blood sugar and plasma insulin responses to oral glucose arising during induction of diabetes are presented. After a two-year period of diabetes, two animals were examined histologically. Lesions in the kidney and retina, similar to early lesions associated with human diabetic nephropathy and retinopathy, were present. Histologic changes related to the diabetes were also seen in the pancreas and pituitary. The means of induction of the diabetes is discussed. The study supports the view that the dog is a useful species in which to study the long-term pathology of diabetes,


HL-A system and diabetes mellitus.

HL-A system and diabetes mellitus. HL-A antigens were determined in 100 patients with diabetes mellitus. When the data are combined with that from other studies, there is a definite positive association of acute-onset juvenile diabetes mellitus with HL-A8 and W15. Four families are described in which two or more members with this type of diabetes are present, and in each family, affected individuals share a haplotype including HL-A8 or W15. These findings are consistent with the possible role of immune response genes in the HL-A chromosomal region which might control the immune response to virus infections capable of producing islet cell damage.


[Alpha-1 fetoprotein in maternal serum (author's transl)]. Serum alpha-1 fetoprotein (AFP) had been determined by radioimmunoassay in 578 patients. During normal pregnancy the AFP concentration increases continuously till 32nd week of pregnancy while thereafter a decrease has been observed. Values of AFP which were found beyond standard deviation indicate fetal distress. Further studies will contribute to the significance of AFP for monitoring normal pregnancies, determination of gestational age and diagnosis of fetal distress in cases of high risk pregnancies.


Serum antigens of cattle. IV. Immunogenetics of a soluble antigenic determinant derived from lysed erythrocytes. This paper describes an allotypic system in cattle called Ec (Erythrocyte cattle). The antigenic determinant is derived from red cells and is detected in lysates by gel precipitation with alloimmune serum. The Ecl specificity is controlled by an autosomal dominant gene and occurs with variable frequencies in different cattle breeds. The Ec system is a very useful genetic marker because the homozygous and heterozygous genotypes show a dosage effect and can be distinguished by the size of the precipitin rings in double diffusion gel precipitation tests.


On the geographical distribution of pseudocholinesterase variants. The incidence of pseudocholinesterase (PCHE equals E.C. 3.1.1.8) variants in samples of 8 different population (total of 2218 individuals) is reported. Together with previously mentioned data from the literature, a general survey on the geographical distribution of PCHE isoenzymes is given. Possible reasons for present-day heterogeneity of their distribution are also discussed. Concerning the incidence of the C5 variant, it is pointed out that the validity of applying population genetic models depends upon the accuracy of the genetic basis.


The value of fluorescence markers in the distinction between maternal and fetal chromosomes. Selected fluorescence markers of chromosomes were studied in 50 paired samples of cells obtained by culture of amniocentesis material and by culture of leukocytes from pregnant women. Comparative analyses showed that this method is of great value in disclosing admixture of maternal cells to material obtained by amniocentesis, as a minimum of 2 fluorescence marker differences between mother and fetus was found in the present material. The distribution of markers in mother/fetus pairs is in agreement with the assumption of genetic determination. Variation was observed between populations with respect to the frequency of the markers studied.


Observations on specific giemsa staining of the Y and on selective oil destaining of the chromosomes. The human Y chromosome can be differentially stained with Giemsa using simple procedures. This phenomenon is strikingly to that observed with quinacrine fluorescence. The specific Giemsa-Y stain may be selectively removed by the action of an oil. The same oil, under certain conditions, selectively removes Giemsa stain from all chromosomes, resulting in R- and T-banding patterns. These bands, which are obtained through subtraction of dye from Giemsa-stained chromosomes, allow slides to be further processed.


Interaction of the hinge region of human immunoglobulin G with a murine lymphocyte membrane receptor. Relevance to the problem of antiglobulin induction in rheumatoid arthritis. Evidence is presented which indicates the presence on murine lymphocytes of a membrane receptor for determinants on the hinge region of human IgG. These determinants are exposed following pepsin scission of IgG molecule, i.e. on the F(ab')2 fragment. The evidence for a hinge receptor derives, in vivo, from splenic localization of F(ab')2 in germinal centres and, in vitro, from immunofluorescent binding studies. The sequential immunofluorescent pattern for the uptake of human F(ab)2 fragments into murine spleen germinal centres was identical with that previously observed for heat-aggregated human IgG, but F(ab')2 fragments appeared to be retained in the germinal centres for a shorter time than aggregated IgG. Experiments with nude mice and T cell-deprived mice showed that the localization of F(ab')2 fragments does not require T cells. Competition experiments suggest that the receptor for F(ab')2 may bear little relation to the receptor for aggregated IgG. The relevance of such a lymphocyte membrane receptor to the immunopathology of rheumatoid arthritis is discussed in the light of previous findings that a proportion of the serum IgG of patients with rheumatoid arthritis has a structural anomaly compared with control IgG, characterized exposure of new determinants at the hinge region.


Surface immunoglobulin of guinea-pig leukaemic lymphocytes. The surface immunoglobulin of the transplantable L2C leukaemia of strain 2 guinea-pigs has been investigated. The immunoglobulin is seen to be synthesized when the cells are maintained in culture, indicating its intrinsic origin. Immunolabelling of the cell surface and immunochemical study of the Fab released by limited surface proteolysis indicate the presence of immunoglobulin of class IgM. IgG and free light chains were not detected, and there is unlikely to be an appreciable amount of immunoglobulin of any other class. The amount of immunoglobulin present, in terms of 4-chain monomers, is approximately 100,000 molecules per cell. Its half-life, calculated from the rate of reappearance in vitro of surface Fab after proteolytic clearing, is approximately 5 hours. Immunoglobulin secreted into the environment appears to arise predominantly or entirely from the cell surface: there is no evidence of an appreciable export of immunoglobulin which does not have a surface phase. Papain at 0.06 mg/ml rapidly removes the surface Fab. Residual Fcmu can then be detected by immunofluorescence, suggesting that papain cleaves surface IgM at a hinge region with the molecule in situ on the membrane. The released Fab is only moderately susceptible to degradation by papain at the enzyme: substrate ratio prevailing. It has been possible to isolate it from the papain digest by immuno-adsorption, with a notional yield of 75 mug per 10-10 cells, and then to prepare antisera against it.


Development of surface immunoglobulins in the chicken. B-cell maturation in the chicken has been evaluated by the appearance of membrane immunoglobulins on cells in the spleen, the thymus, the bursa and the bone marrow during enbryonic development and shortly after hatching. The majority of the bursa cells acquire demonstrable membrane immunoglobulin between days 16 and 18 of incubational age and show significantly increased amounts of membrane immunoglobulin between days 18 and 20, even though immunoglobulin-bearing cells can be found in the bursa as early as day 14 of enbryonic age. The spleen shows cells possessing immunoglobulin receptors can their membranes (Ig+) only after the bursa cells have reached full membrane immunoglobulin maturation as reflected in the number of Ig+ cells and the amounts of membrane immunoglobuline. The thymus is practically devoid of Ig+ cells in the embry and it is not clear whether there any Ig-+ cells in the bone marrow. There are two phenomena which stand out in the observations. One is that there appears to be a gradual increase in the quantity of quality of the surface immunoglobulins on individual cells with advance in the embryonic development as reflected in the gradual increase in the staining intensity. The other is that there appears to be a polar distribution of membrane immunoglobulin in some cells especially in younger embryos. This polar distribution is seen under conditions where immunoglobulin capping is prevented by inhibitors and where immunoglobulin capping is impossible, such as with monomeric Fab. Immunoglobulin capping has been found to occur readily in embryonic cells and under conditions which would normally inhibit capping in adult cells.


Importance of short-lived lymphocytes in the immune response. Lymphocytes are heterogeneous with respect to their life-span. Typical B cells, bearing on their membranes immunoglobulin receptors, easily detectable by immunofluorescence, belong mainly to the long-lived population: this can be observed using combined autoradiography and immunofluorescence. However, when primed mice receive (-3H) thymidine before a boosting injection of tobacco mosaic virus (TMV), many plasma cells appearing in the spleen during the secondary response are labelled. In irradiated recipients repopulated with spleen cells from donors primed with TMV and injected with tritiated thymidine 2 hours before killing, the majority of plasms cells appearing in the spleen after antigen injection were labelled. If irradiated mice were repopulated simultaneously with spleen cells from donors primed with TMV and injected with (-3H) thymidine, and from donors primed with haemocyanin, most of the anti-TMV plasms cells were labelled, while most of the anti-haemocyanin plasma cells were unlabelled. These results allowed us to exclude non-specific reutilization of labelled thymidine as the main reason of our observations. It is concluded that either plasma cells derive from shortlived precursors or they receive material from a labelled cell able to co-operate specifically with plasma cell precursors.


Antibody formation in the mouse induced by hapten-carrier complexes. The influence of hapten, carrier and their ratio in a complex on T-cell helper stimulation and antibody formation against the dinitrophenyl (DNP) hapten was studied. Complexes of DNP with bovine serum albumin (BSA), bovine gamma-globulin (BGG), isologous mouse immunoglobulin (MIG) and polyvinylpyrrolidone (PVP) as carriers were used. Optimal antibody formation against DNP was obtained with complexes with an intermediate hapten:carrier ratio (DNP 16-minusBSA, DNP 43-minusBGG and DNP 48-MINUSMIG). DNP-PVP complexes were not active either in the primary or in the secondary response. The anti-BSA titre was independent of the number of DNP groups on the complex used for immunization. Inhibition of DNP-plaque formation by spleen cells of immunized mice shows an increase of the inhibitory capacity of the complex with the increase of the hapten-carrier ratio. DNP 16-minusPVP was the only PVP complex which was inhibitory. These results suggest that helper cells involved in the antibody formation against BSA and DNP are reactive with different parts of the complex. Priming of mice with carrier or complex after cyclophosphamide (Cy) treatment followed by a secondary injection with complex 10 days later gave strong indications that there is a greater involvement in stimulation of helper T cells by determinants of the complex (new antigenic determinants (NAD) or NAP-DNP groups) or DNP, than by true BSA determinants. This holds for both the IgM and IgG responses.


Delayed hypersensitivity in the mouse induced by hapten-carrier complexes. Delayed hypersensitivity (DH) in the mouse was studied with complexes of dinitrophenyl (DHP) as hapten and bovine serum albumin (BSA), mouse immunoglobulin (MIg) and polyvinylpyrrolidone (PVP) as carrier. Priming with BSA induced strong DH against this carrier, but DN of decreasing strength against complexes with increasing DNP:carrier ratio. Priming with DNP-BSA complexes never resulted in a DH against BSA or DNP 3-minusBSA. Injections of DNP 16-minusBSA and DNP 28-minusBSA induced positive DH which increased with the hapten:carrier ratio of the eliciting antigen. The complexes with an isologous carrier DNP 48-minusMIg or DNP 90-minusMIg induced positive reactions against both complexes but not against the weakly substituted DNP 11-minusMIg. The latter only primed for itself. The importance of the DNP groups as determinant in these DH reactions is stressed by the cross-reactions between DNP-BSA and DNP-MIg complexes and by the induction by DNP 16-minusPVP of positive DH against DNP 28-minusBSA. Cyclophosphamide (Cy) treatment before priming with complexes induced enhanced DH against complexes with sufficient hapten:carrier ratio. Priming with carrier under Cy treatment induced no DH against complexes. All these results indicate that carrier determinants are not involved in the DH against complexes. After priming wtih complexes with a low hapten:carrier ratio the DH is directed against new antigenic determinants (NAD) groups. After priming with complexes with high ratios DH is directed against DNP groups. With adoptive local transfer of spleen cells of primed animals and pretreatment of these cells with anti-thymocyte serum or anti-plasma cell serum and complement it was possible to demonstrate that the T cell was responsible for the DH reactions. The involvement of different determinant groups on the hapten-carrier complexes in immune reactions is discussed.


Hypersensitivity to mineral oil in the domestic fowl. Domestic fowl were found to be hypersensitive to a second injection of highly purified mineral oil if they had been primed 3 weeks previously with oil in the presence of mycobacteria. This response did not appear to be related to impurities originally present in the oil or introduced into it during sterilization, and did not follow priming with mycobacteria in vegetable oil or in saline. The reaction at the challenge site was prolonged and developed a characteristic histology identical to that found at the priming site. It is postualted that the hypersensitivity seen has an immunological basis and that birds are able to respond to antigenic determinants formed by long chain aliphatic hydrocarbons.


Rat tumour allografts evoke anaphylactic antibody responses. In vitro assays of release of histamine from peritoneal mast cells showed that Wistar rats produced anaphylactic antibody in response to a single immunization with an allogeneic sarcoma. The response occurs early after immunization, and no adjuvant is needed. The thermolability of the anaphylactic antibody suggests that it is IgE.


Release of histamine from rat mast cells by the complement peptides C3a and C5a. Suspensions of rat mast cells were used to study the histamine-releasing actions of anaphylatoxins C3A and C5a in vitro. The peptides, derived from human or porcine complement proteins C3 and C5, were less potent than 48/80 but more potent than bradykinin in stimulating release of histamine from mast cells. The pattern of release resembled that of the anaphylactic release action, e.g. release was limited to less than 30 per cent of the cell histamine, the reaction was calcium-dependent and was potentiated by phosphatidyl serine. When C3a and C5a were added together to mast cell suspensions, the amount of histamine released was additive. Similarly, release by either peptide combined with bradykinin was additive. Histamine-releasing activity (as well as smooth muscle-stimulating activity) was abolished when the peptides were treated with pancreatic carboxy-peptidase B. Active or inactive peptides were bound by mast cells and addition of active C3a in combination with the inactive, des-arginine derivative, C3ai, resulted in partial inhibition of histamine release.


Inhibition by serum of encephalitogenic activity of myelin basic protein. Basic protein of myelin from bovine brain (B-BPM) in Freund's complete adjuvant (FCA) is highly encephalitogenic for the guinea pig. However, when B-BPM is mixed with serum from various species it loses its encephalitogenic effect but not its immunogenic properties. When the synthetic tryptophan peptide matching residue 115-126 of human BPM is mixed with normal human serum it loses both its encephalitogenic and immunogenic effects. The time of exposure to serum necessary for complete inhibition of encephalitogenic activity of B-BPM varied: rat, horse, sheep and human sera produced their inhibitory effect immediately after mixing, whereas guinea pig serum required 8h. The capacity of serum to abrogate the encephalitogenic effects of B-BPM was not due to complete degradation of the molecule by proteinases in serum, because all animals injected with the B-BPM serum mixture gave cutaneous delayed hypersensitivity reactions to B-BPM. The inhibitory effect could be attributed to a factor in serum which is non-dialysable, thermostable at 56 degrees C, for 1 h, and present in high concentration in fetal calf serum. It may be an alpha2 macroglobulin which can act selectively on the main encephalitogenic determinant, around or within residues 115-126 of the basic protein of myelin.


Modulation of the immune response by passive antibodies. I. Anti-hapten antibodies enhanced delayed hypersensitivity to the carrier and depressed antibody synthesis to the hapten. The modulating effects of passive antibodies on both delayed hypersensitivity to the carrier and antibody synthesis to carrier and hapten determinants were studied in guinea pigs. Animals were injected with antibodies directed against either the carrier or the hapten prior to immunization with the hapten-carrier conjugate in Freund's complete adjuvant. Anti-hapten antibodies have been shown to have an enhancing effect on delayed hypersensitivity to the carrier and a suppressive effect on antibody synthesis to the hapten. In this experiment, anti-carrier anti-bodies seemed to have had no effect on delayed hypersensitivity to the carrier and on antibody synthesis to the hapten.


Intraaticular treatment of tarsal degenerative joint disease in cattle. Tarsal degenerative joint disease (DJD) in 12 cattle was classified as primary or secondary, based on age, evidence of hereditary or congenital joint conformation defects, faulty hindlimb alignment, duration and type of usage joints were subjected to, and history or signs of repeated trauma. Three of the cattle had bilateral primary tarsal DJD, 7 had bilateral secondary tarsal DJD, and 2 had secondary DJD of the left tarsus. Analyses of synovial fluid samples provided a means of characterizing pathologic changes of tarsal DJD, Results of blood and synovial fluid analyses were grouped in compilation of data for cattle affected with either primary or secondary tarsal DJD. Corticosteroids and a long-acting synthetic progestational agent were injected singly or in combination with aqueous antibiotics into affected tarsal joints. Tarsal joints of 5 of the cattle responded favorably to a single intraarticular treatment, as manifested by palliative relief and functionally usable joints. Seven joints of 5 cattle were subjected to repeated intraarticular treatment. Serial synovial fluid analyses in 7 of the cattle provided a means of assessing tarsal joint response to intraarticular treatment or to therapeutic arthrocentesis, exclusive of patient objective response. One cow developed a mild self-limiting bilateral postinjection synovitis that was resolved after the 2nd and final intraarticular injection. Usable function returned to tarsal joints of cattle that responded favorably to intraarticular treatment at different periods after single or repeated injections. Three cattle with advanced tarsal DJD experienced minor temporary relief and were euthanatized at their owner's request. Improvement did not occur in the tarsal joint of 1 cow subjected to therapeutic aspiration only. Intraarticular treatment in all cattle was considered supportive to the animal's well-being rather than curative.


Mucosubstances in Brunner's glands of the mouse. The mucins of Brunner's glands in the mouse have been studied by histochemical methods. The secretion droplets in both acinar cells and the cells lining the ducts are PAS-positive and diazyme-resistant. The latter cells are alcianophilic at pH 2.5, react positively with Hale's colloidal iron method and exhibit metachromasia with azure A. The reaction to Hale's colloidal iron method is lost after digestion with neuraminidase. These results suggest that an acid as well as a neutral mucopolysaccharide is present in the secretion of Brunner's glands of the mouse, the acid mucosubstance being a sialomucin. The possible significance of this finding is discussed. This work was performed in partial fulfilment of work for the degree of Doctor of Philosophy of the University of Sheffield. It was supported by a Research Fellowship from the World Health Organisation (W.H.O.) during a study leave granted by the University of Lagos, Nigeria.


Computer analysis of organelle translocation in primary neuronal cultures and continuous cell lines. Organelle translocation in a number of cell types in tissue culture as seen  by high-resolution Zeiss-Nomarski differential interference contrast optics was filmed and analyzed by computer. Principal cell types studied included primary chick spinal cord, chick dorsal root ganglion, ratbrain, and various clones of continuous cell lines. Organelle translocations in  all cell types studied exhibited frequent, large changes in velocity during any one translocation. The appearance of particles as seen with Nomarski optics was correlated with their fine structures in one dorsal root ganglion neurite by fixing the cell as it was being filmed and obtaining electron micrographs of the region filmed. This revealed the identity of several organelles as well as the presence of abundant neurotubules but no neurofilaments. Primary cell cultures exhibited more high-velocity organelle movements than continuous cell lines. The net progress of an organelle in a given direction was greater in primary neuronal cells than in fibroblasts or continuous cell lines. These findings are correlated with the literature on organelle translocation and axoplasmic transport.


Characterisation and quantitation of morphine in urine using high-pressure liquid chromatography with fluorescence detection. A simple and rapid method for the identification and quantitation of morphine in urine samples is described. The procedure, which involves conversion of the drug a fluorescent product followed by liquid chromatography, is shown to be highly sensitive and specific. Levels down to 0.01 mug/ml of morphine can be quantitatively detected in urine. A large number of drugs have been tested and shown not to interfere.


Retrograde axonal transport of horseradish peroxidase in peripheral autonomic nerves. An exogeneous marker protein, horseradish peroxidase (HRP) was used to race peripheral autonomic pathways in adult guinea pigs and cats. Small doses of HRP were injected into various organs and after a brief survival period, HRP activity appeared in the perikarya of autonomic neurons that supplied each injection site. After injection of HRP into the anterior chamber of the eye, reaction product was detected in the postganglionic sympathetic neurons of the superior cervical sympathetic ganglion. In another experiment, HRP reaction product was found in the cell bodies of the preganglionic sympathetic neurons that supply the adrenal medulla. These were located in the lateral gray column of the spinal cord at T6 and T7 segmental levels. Reaction product appeared in intramural postganglionic parasympathetic neurons close to an injection site in the wall of the urinary bladder and in a similiar situation in Meissner's ganglia of the ileum. Following injection into the walls of the stomach and ileum, HRP labelled cells were detected in the nodose ganglion of the vagus and in preganglionic parasympathetic neurons in the dorsal motor nucleus of this nerve. After injection into the subepicardial tissue of the heart, reaction product appeared in the stellate ganglion and also in an upper thoracic dorsal root ganglion. These data suggest that HRP is taken up by peripheral autonomic nerves of all types, and then undergoes rapid retrograde axonal transport to the perikaryon. It appears, therefore, that HRP may be useful in tracing both motor and sensory peripheral autonomic pathways.


Locus ceruleus in rhesus monkey (Macaca mulatta): a combined histochemical fluorescence, Nissl and silver study. The locus ceruleus (LC) of the rhesus monkey (Macaca mulatta) was investigated using the histochemical fluorescence method and Nissl and ammoniacal silver stains. Caudally, a few fluorescent cells were observed in the lateral wall of the ventriculus quartus near the velum medullaris superior. Rostrally, the fluorescent cells were compactly clustered and reached their greatest density medial and, to a lesser extent, lateral to the tractus mesencephalicus n. trigemini at the level of the decussation of the nervus trochlearis. In the most rostral plane, fluorescent cells were more diffusely scattered ventral and medial to the pedunculus cerebellaris superior with a few cells situated near the nervus trochlearis. Nissl staining of tissue previously used for histochemical fluorescence showed that fluorescent cells were largely found in a region labelled LC in two rhesus monkey brain atlases and erroneously labelled nucleus tractus mesencephalicus n. trigemini in one brain atlas. Ammoniacal silver staining resulted in a dense accumulation of silver granules in the cells found to display a positive reaction for monoamines. The silver stains therefore offer an alternative to the histochemical fluorescence method for identifying the monoamine-containing LC neurons.


In vivo studies of mediator release in cold urticaria and cholinergic urticaria. Six patients with cold urticaria were found to possess elevated plasma histamine levels after cold challenge by placing one hand in ice water for 4 minutes. A single patient became hypotensive during the procedure and had a level of 260 ng/ml. histamine in the venous effluent from his hand. No elevation of plasma serotonin or bradykinin was observed. Two patients with cholinergic urticaria possessed elevated plasma histamine levels during and after vigorous exercise for 10 minutes; these patients also gave a positive test for vibration-induced angioedema. A single patient with cholinergic urticaria possessed elevated baseline serotonin levels and elevated levels during and after exercise but no elevation of plasma histamine or bradykinin. The results suggest that histamine is the major mediator of urticaria and hypotension in cold urticaria. Histamine also appears to be released coincident with the development of urticaria in some patients with cholinergic urticaria, while elevated serotonin levels in a single atypical patient suggest that a subpopulation of patients with cholinergic urticaria possess a different pathogenesis.


Inuction of immunological tolerance in immunoglobulin E B lymphocytes in rats. Immunological tolerance has been induced in 2,4-dinitrophenyl (DNP)-specific bone marrow-derived or B lymphocytes of the IgE AND IgG antibody classes by treatment of rats with the DNP derivative of D-amino acid copolymer, D-glutamic acid, D-lysine (D-GL). The tolerant state is manifested as an inability of treated rats to produce serum anti-DNP antibodies and the failure of peritoneal cells from tolerant animals to release histamine following in vitro antigen challenge. The implications of these and related observations for potential therapeutic measures in clinical hypersensitivity states are discussed.


Properties and activities of transfer factor. Although there is agreement that transfer factor endows skin test-negative subjects with the ability to develop the delayed allergic responses of the transfer factor donors, there is little direct information on the mechanism of this phenomenon or on the nature of the active components (s). This report reviews some of the known effects of transfer factor or immune responses and inflammation. It is concluded that transfer factor has multiple sites of action, including effects on the thymus, on lymphocyte-monocyte and/or lymphocyte-lymphocyte interactions, as well as direct effects on cells in inflammatory sites. It is also suggested that the "specificity" of transfer factor is determined by the immunologic status of the recipient rather than by informational molecules in the dialysates. Finally, it is proposed that many effects of transfer factor may be due to changes in intracellular cyclic nucleotide content, especially accumulation of cGMP, in immunologically reactive cells.


The effect of Tris-buffered media and Tyrode physiologic saline solution on the antigenic release of histamine from human leukocytes. Allergic histamine release from leukocytes was compared in three different media: Tyrode physiologic saline solution, Tris-buffered saline containing human albumin, calcium, and magnesium (Tris-ACM), and Tris-ACM with homologous serum. In a selected group of low histamine releasers, the maximal amount of antigenic histamine release was significantly higher in Tyrode solution as compared to Tris-ACM buffer. When homologous serum was added to Tris-acm, an enhancement of histamine release greater than with Tyrode solution was obtained. These results suggest that Tris-ACM may not be the optimal buffer for leukocyte histamine release experiments. Since Tyrode solution contains no serum proteins that bind slow-reacting substance of anaphylaxis or prostaglandins, the use of this medium may be advantageous for the study of the release of the chemical mediators from human leukocytes.


Primary interaction between antibody and components of Alternaria. I. Immunology and chemical characteristics of labeled antigens. Components were isolated from Alternaria tenuis and its culture filtrate, and were radiolabeled with 123-I. The labeled antigenic components had a high polysaccharide content as determined by staining patterns following electrophoresis in polyacrylamide gels and by inactivation with sodium metaperiodate. A primary binding test was employed to detect and measure serum antibodies to the components from A. tennuis. This procedure was more sensitive in detecting antibodies that bound to antigens than were comparable tests dependent upon precipitin types of reactions. The labeled components of A. tenuis cross-reacted or shared antigens with 3 other species of molds: Stemphylium sp., Curvularia sp., Aspergillus fumigatus, but not with a variety of other fungal and nonfungal materials.


Group psychology and the structural theory: a revised psychoanalytic model of group psychology. It has been my aim in this paper to revise the existing psychoanalytic theory of group psychology in accordance with current structural concepts. The need for fundamental revision in the existing theory of group organization is demonstrated by its restriction to an oedipal paradigm, which cannot account for the regression to an oral paradigm of group organization during group formation. Freud's explanation of regression in crowds is reviewed. The limitations inherent in Freud's topographic and narcisistic models are demonstrated; irreconcilable contradictions are shown to exist between the two theories. A structural model of group psychology that is free from internal contradictions and provides a unifying explanation for both regression and merging in the crowd is developed. As a consequence of these revisions it is possible to conceptualize preoedipal organizations of group structure in addition to the oedipal paradigm proposed by Freud.


Experimental hematogenous endophthalmitis caused by Candida albicans. For a rabbit model of hematogenous endophthalmitis caused by Candida, 80 New Zealand white rabbits were given intravenous injections of 10-5 Candida albicans. Two weeks after infection 88 percent of infected rabbits had ocular lesions with clinical appearance identical to that of human lesions. Cultures of the chorioretina and kidneys averaged 10-4.3 and 10-3.9 organisms per g of tissue, respectively, three days after infection and remained positive for 38 days. Ninety-five percent of rabbits with positive cultures of the chorioretina had positive cultures from other organs. The ocular lesions were focal chorioretinitis characterized by a combination of granulomatous and acute suppurative reactions. Hematogenous endophthalmitis caused by Candida in the rabbit is a useful model for study of the pathogenesis and treatment of the disease in humans.


Experimental pneumonitis and encephalitis caused by acanthamoeba in mice: pathogenesis and ultrastructural features. For a more precise definition of the clinicopathological features of experimental acanthamoebic infection in mice, trophozoites of Acanthamoeba castellanii and Acanthamoeba polyphaga were instilled intranasally into adult white mice. Eight to 20 days after inoculation, severe pulmonary disease developed; one to two days later, neurological signs ensued. On pathologic examination an amebic broncho-pneumonia associated with encephalitis was found. Trophozoites and cysts were seen in lung and brain. Although Naegleria is spread by the olfactory route, cerebral lesions produced by Acanthamoeba might result principally from hematogenous carriage from the lungs. Other differences between infections caused by Naegleria and those caused by Acanthamoeba in mice also exist and serve to emphasize that when natural infections with Acanthamoeba occur, a distinct clinicopathological entity may be produced.


Subtypes of hepatitis B surface antigen in Southeast Asia. The prevalences of hepatitis B surface antigen (HBs Ag) subtypes in Thais, Cambodians, and Vietnamese were compared with the prevalences in Americans residing in Southeast Asia. HBs Ag was found with approximately equal frequency in Thai (43 percent) and American (39 percent) patients with hepatitis. However, higher prevalences of HBs Ag were found in asympotomatic Thais (9.5 percent), Cambodians (11.9 percent), and Vietnamese (14.3 percent) than in asymptomatic Americans (0.7 percent). Among asymptomatic Thais, the ratio of HBs Ag/adr to HBs Ag/adw was approximately 10:1, with one exception: adw was not detected in a rural population of northern Thailand. The y determinant was not found in Thais. In contrast, both d and y determinants were found in Americans. These observations conform to a geographic pattern, with ad as the predominant combination in the Far East. In Southeast Asia determinants w and r are more useful epidemiologic markers than y and d.


The relationship of thyroxine turnover to other parameters of peripheral thyroid function in Graves' disease. In 28 patients with Graves' disease showing a wide range of thyroid function between the extremes of hypothyroidism and hyperthyroidism, the following parameters of peripheral thyroid function were measured: serum thyroxine concentration, serum-free thyroxine concentration, serum triiodothyronine concentration, and serum-free triiodothyronine concentration. In 25 patients, thyroxine turnover was also measured. Thyroxine turnover was found to be highly correlated with serum-free thyroxine concentration (r equals 0.9405) and serum-free triiodothyronine concentration (r equals 0.9184). Serum-free thyroxine fraction correlated with serum-free triiodothyronine fraction (r equals 0.8445), suggesting that similar factors in serum controlled the intensity of protein binding for both thyroxine and triiodothyronine. Thyroxine turnover calculated by a noncompartmental method agreed closely with values calculated by the compartmental method, suggesting that the former simpler method has general utility.


Investigation of DNA-content of leukoplakia cells or oral mucosa. DNA-histograms of smears of 11 plain leukoplakias, one of a precancerous leukoplakia, three of leukoplakia carcinoma in early stage, and one of a keratotic seqamous cell carcinoma of the floor of the mouth were prepared. The cell nuclei of the plain leukoplakia are mostly diploid, a small proportion has a DNA-content of 4 C. Intermediate values or polyploid nuclei are absent. In precancerous leukoplakia, intermediate values were noticed; in the cases of carcinoma in early stage, intermediate and polyploid nuclei were present, and in one case only polyploid nuclei. Only 2C and 4C nuclei occurred in a case of keratotic squamous cell carcinoma, which in comparison to earlier examined cases must be termed DNA negative.


Cryosurgical devitalization of bone and its regenreation; An experimental study with animals. In an experimental study using rabbits, bone is supercooled by means of liquid nitrogen flowing through a silicone chamber. The surrounding soft tissue is protected by a heating element of silicone. The histological findings in the stages of devitalization of osteocytes are described. X-rays and fluorescent labelling were used, searching for the time of total recalcification in supercooled bone.


Demonstration of a specific cytosol receptor in the normal and hyperplastic canine prostate for 5alpha-androstane-3alpha, 17alpha-diol. A specific receptor protein has been demonstrated for 5alpha-androstane-3alpha, 17alpha-diol in cytoplasmic extracts of normal and hyperplastic canine prostates. The receptor molecule, with a sedimentation coefficient of 4-5S, has been identified by the use of sucrose gradient centrifugation of tissue fractions which had been labelled in vitro with tritiated 5alpha-androstane-3alpha, 17alpha-diol. The receptor showed a relatively high affinity for this compound whereas binding could not be demonstrated with other labelled C19 steroids. In addition binding of tritiated 5alpha-androstane-3alpha, 17alpha-diol was not affected by the presence of 50-fold excesses of other C19 steroids.


Influence of antigen dose on antibody production of intact and splenectomised Xenopus laevis. Antigen persistence and serum antibody production in intact Xenopus were monitored using human gamma globulin (HGG), in adjuvant, in various immunisation schedules. Retention of HGG in spleen and serum was directly related to the quantity injected. However, antibody responses to a dose range between 1 mu-g-6 mg antigen were similar in intensity. These were detected in the serum two weeks after injection and at this stage were exclusively mercapto-ethanol (ME) sensitive; ME-resistant antibodies had appeared by four weeks. No antibodies were detected below a dose of 100 ng HGG. The effect of splenectomy on antibody levels was tested using HGG/adjuvant or sheep erythrocytes (SRBC) in saline. Splenectomised toads showed impairment of antibody responses only to threshold doses of HGG (100 ng) but to a wider range of SRBC doses.


Salmonella typhimurium mutations conferring resistance to Felix O phage without loss of smooth character: phage attachment and immunochemical and structural analyses of lipopolysaccharides. Salmonella typhimurium mutants, called Felix O-resistant (FOR), selected for resistance to phage Felix O (FO) which has its receptor in the core lipopolysaccharide (LPS), retain most of the properties of the smooth parent strain (MacPhee, Krishnapillai, Roantree & Stocker, 1975). LPS extracted from one parent and two FOR strains by the phenol-water and the phenol-chloroform-light petroleum methods have been subjected to passive haemagglutination inhibition and methylation analysis. The amount of LPS, the amount of O-specific sugars in the LPS, and the average length of the O chains were almost the same in parent and mutant strains. Neither passive haemagglutination nor methylation analysis revealed the presence of incomplete cores in the mutant strains. Determination of the rates of attachment of P22 (receptor in O chain) and FO phages to whole bacteria of the same strains also suggested there is as much O-chain material in the FOR strains as in the parent strain. The data suggest that the FOR strains are the result of a mutation in the synthesis of the core, leaving few, if any, completed cores accessible to the FO phage.


Esterase zymograms of Proteus and Providencia. The intracellular esterases of 80 strains of Proteus and Providencia were analysed by the acrylamide-agarose zymogram technique using several synthetic substrates. The esterase bands were classified in five main groups. The alphaA-esterase bands hydrolysed alpha-naphthyl acetate and were resistant or relatively insensitive to di-isofluoropropyl phosphate (DFP). The alphaB-esterase band hydrolysed both alpha-naphthyl acetate and alpha-naphthyl butyrate and were very sensitive to DFP. Both groups of esterase bands were inactivated by heat. The betaA- and betaB-esterase bands hydrolysed beta-naphthyl acetate and were sensitive to DFP; these were distinguishable by the difference in their relative activity towards beta-naphthyl butyrate and in their relative stability to heat. The alpha-beta-esterase bands hydrolysed alpha- and beta-naphthyl acetates and alpha- and beta-naphthyl butyrates; they were inactivated by heat and were sensitive to DFP. The distribution of these esterase bands among the strains of Proteus and Providencia and their electrophoretic patterns established esterase profile types which correlate with the classification based on traditional bacteriological tests. The degree of inter-strain similarity in esterase pattern varied highly among species. The homogeneity of Proteus mirabilis and especially of Providencia stuartii contrasted with the heterogeneity of other species. This disparity suggests that the bacteria of the tribe Proteae have not the same degree of intra-specific differentiation in physico-chemical properties of esterases.


Astrocytic processes: A route between vessels and neurons followingblood-brain barrier injurt. The fluorescent tracer Evans blue (EB) was studied after circulation time ranging from 30 seconds to 5 minutes following blood-brain barrier (BBB) injuries in 42 dogs. Selective neuronal stainings (sns), associated with  BBB injuries, occurred to a high degree. This phenomenon was examined by modifications in existing fluorescence techniques. The mechanism of SNS was seen to lie in an almost imperceptible transport of EB along astrocytic processes connecting SNS with the adjacent vasculature. This report presents visual evidence under pathological conditions, and supports the concept of astrocytic processes as the normal route of transport between blood vessels and neurons. This requires long exposure photomicrographs since the fluorescing material is subtle within the neuroglial processes.


Occurrence and natural history of chronic lymphocytic thyroiditis in childhood. In a six-year survey of 5,179 school children in Arizona, Utah, and Nevada 62 cases of chronic lymphocytic thyroiditis were identified giving a prevalence of 1.2%. Thyroids were enlarged in 85%, firm in 60%, and had an irregular or lobulated surface in 75%. Antibodies to thyroglobulin were demonstrable in the serum at some time during the course of the disease in 76% by the tanned red blood cell technique and in 93% by radioimmunoassay. Serum TSH concentrations were elevated in seven of 15 subjects. Many of the cases were early or mild thyroiditis and, in most instances, subjects were asymptomatic and considered clinically euthyroid. Two subjects were hypothyroid, and two appeared clinically hyperthyroid. Spontaneous resolution of thyroiditis occurred in 15 of 32 individuals who received no treatment. Resolution occurred in 14 of 30 children treated with thyroid hormone supplement. The results suggest that lymphocytic thyroiditis in children may be present without symptoms and in many is a self-limiting disorder from which complete recovery occurs spontaneously.


Thyroid function in term newborn infants with congenital goiter. Eighty-four term newborn infants without goiter and 45 newborn infants with congenital goiter were studied with regard to thyroid function. The radiologic development of the femoral and tibial epiphyses was evaluated in those with goiter. Fifty-eight percent of the patients had retarded bone age, markedly elevated TSH levels, elevated TBI, decreased total T4I, and decreased PBI values. Forty-two percent of newborn infants with congenital goiter had a normal bone age, normal values for TSH, PBI, and total T4I, and elevated values for TBI. It is concluded that the 58% of the newborn infants with congenital goiter had subtle hypothyroidism. They require substitution therapy with thyroid hormones in order to avoid possible retardation of normal brain development. Patients with congenital goiter who have no biochemical evidence of hypothyroidism should also be treated with thyroid hormones to achieve rapid regression of goiter.


Hydrodynamic analog model for pharmacokinetics II: critical examination of model and its contribution to pharmacokinetics. A comparison of the conventional pharmacokinetic models and the previously proposed hydrodynamic diffusion analog model is presented. A significant result that an n-compartment system can exhibit at best (n - 1) extremum times in the concentration-time plot of the central compartment under appropriate values of physiological parameters is presented. The observation of kinks experimentally in certain physiological-drug systems is thus shown to be amenable to explanation.


Polycyclic aromatic isothiocyanate compounds as fluorescent labeling reagents. Polycyclic aromatic isothiocyanates were synthesized in an attempt to produce new fluorescent agents for protein labeling and for use in microanalytical techniques. The relative fluorescence of these reagents, the required intermediates, and the derivatives were established. Of the compounds evaluated, the 9-acridine derivatives were the most promising.


Effectiveness of audio-based instruction in medical pharmacology. An audio-based method of instruction on the topic of diuretic drugs was compared with the lecture method using a random sample of 66 sophomore medical students. A cognitive test given to measure achievement as a result of the two treatments showed that the average score of 80 percent for the audio group was significantly higher (p smaller than 0.25) than the 75 percent for the lecture group. A comparison of the lowest quartile scores in each group revealed that the audio-based group also scored higher 13 points, p smaller than .05) than the lecture group; however, no difference was noted in the upper three quartile scores of the two groups. The amount of time spent studying did not appear to be a factor. A second test given to meausre attitudes toward the audio method showed strong positive views among students.


Radiation damage in stained catalase at low temperature. Radiation damage, which occurs in the beam of the electron microscope, has been studied in uranyl acetate stained crystals. Damage is observed in terms of the disappearance of higher orders of the electron diffraction pattern. In this study it has been found that the damage at very low specimen temperatures proceeds both more rapidly and to a greater final degree than is the case at room temperature.


A symbolic tree: loneliness is the root; delusions are the leaves. This particular case involving the suggested nursing intervention was very successful and rewarding.  It is very important, however, that therapists realize that these seven steps were acceptable in this particular case, but steps four-to-seven may be modified with each couple.  However, the first three steps are more than likely essential to any effective intervention dealing with loneliness.  Aso, the excerpts from this case happen to illustrate the highlights that were related to loneliness.  The therapists recognized that there were other issues and concerns that also needed to be dealt with and which were, but the need for intimacy seemed to have top priority.  Loneliness rooted in the symbolic tree cannot be dealt with by plucking the leaves; it must be dug out by the roots.  Psychotherapy needs nursing psychotherapists who will contribute to this digging.


Studies of parents of children with acute leukemia. In a case-control study, 70 mothers and 24 fathers of children with acute leukemia (AL) were compared with 70 mothers and 24 fathers of normal children. Three significant differences (p smaller than 0.05) were found when 35 factors were compared among the mother pairs and one difference among the father pairs. Mothers of children with AL, though alike in most respects to their matched controls, had a significantly lower number of monocytes than their controls. This was a new observation. The mothers of the children with AL also had higher levels of gamma-globulin, IgA, and IgG (Philadelphia only), which confirmed previous observations. The fathers and mothers had higher levels of basophils. These findings direct attention to the immune systems, particularly the mononuclear cells, of the parents of children with AL, as a focus for further studies on the etiology and pathogenesis of childhood leukemia.


Alpha fetoprotein: effect of heterologous antiserum on hepatoma cells in vitro. Hepatoma cells derived from The Jackson Laboratory mouse hepatoma BW7756 synthesized alpha fetoprotein (AFP) in vitro. The AFP was immunologically identical to that circulating in the sera of hepatoma-bearing mice. An in vitro cytotoxic effect of rabbit antiserum to AFP was studied in hepatoma cells obtained both from fresh cell suspensions and short-term cell culture. The use of intact and/or inactivated anti-AFP serum inhibited the growth of the AFP-producing cells. The cytotoxic effects of the antiserum depended on exposure time and serum concentration. The cytotoxicity was complement independent, as demonstrated by studies with heat-deactivated serum devoid of extrinsic complement. The control target cells included fresh cell suspensions of normal mouse liver and mouse muscle fibroblasts grown in short-term culture. Specificity of the antisera for the target cells was demonstrated by absorption with purified mouse AFP. The results could be explained by the presence of AFP on the hepatoma cell surface.