Detection of suspicious cells and rejection of artefacts in cervical cytology using the Leyden Television Analysis System. In slide based automation of cervical cytology the first stage of analysis involves finding possibly suspicious cells, or areas on the slide with these types of cells. By using a television based system such as the Leyden Television Analysis System (LEYTAS), a number of detection methods can be applied to rapidly screen a large number of fields automatically for suspicious cells. In this paper, results using a parameter based on increased nuclear DNA content of cells are given and a second detection method based on a chromatin pattern feature, called chromatin contrast, is discussed. Two blind trials on 41 positive and 22 negative cervical slides, using the Leyden Television Analysis System to detect suspicious cells with an increased nuclear DNA content, were promising. In 1 of the 41 positive cases no suspicious cells were found. In the negative specimens, suspicious cells were detected in 1 of 9 cases and 1 of 13 cases, with the two detection parameters investigated. These findings are discussed and some automatic artefact rejection procedures with preliminary results are given.


BioPEPR: a system for the automatic prescreening of cervical smears. A feasibility study has indicated that a Prescion Encoding and Pattern Recognition (PEPR) cathode ray tube prescreening system for cervical smears can be both accurate and fast. Smears are prepared using a syringing technique and are stained with a Feulgen-type nuclear stain and a protein counter-stain. The use of film as an intermediate step between the cells and Bio PEPR allows the scanning of fields as large as 8 x 8 mm. The morphological features of the cells are measured as directed by a hierarchical decision strategy. Additional programs detect artifacts, overlaps, and leukocytes. For clean samples, false positive and false negative rates on the cell level have been obtained that will allow acceptable smear level rates (10% false positive, 1% false negative). These rates have been reached without compromising the required speed goals of 120 to 180 smears per hr. The efficiency of the system is dependent on the quality of the smears. Measurements on a set of 192 routinely prepared smears indicate acceptable false negative rates and a false positive rate of about 18%. A reduction of this rate is expected with small improvements in cell preparation and measuring software, leading to the overall system efficiency required for commercial feasibility.


Pairs of fluorescent dyes as probes of DNA and chromosomes. If two fluorescent dyes with different binding or fluorescence specificities are used simultaneously to stain DNA or chromosomes, the ratio of their fluorescent signals can provide information about base composition or base analogue substitution. Energy transfer between such dye pairs, possible if the fluorescence spectrum of one overlaps the absorption spectrum of the other, can modify observed fluorescence. Microfluorometric measurements were used to document the occurrence of energy transfer between quinacrine or 33258 Hoechst as energy donor and ethidium or 7-aminoactinomycin D as acceptor when used jointly to stain cytologic preparations of human metaphase chromosomes. Use of 7-aminoactinomycin D, a dye with G-C binding specificity, as energy acceptor permitted the identification of human chromosome regions presumptively enriched for clusters of A-T base pairs, based on the resistance of A-T specific fluorescence, from quinacrine or 33258 Hoechst, to energy transfer dependent quenching. The results provide information about basic structural features of metaphase chromosomes, and the associated methodology may prove useful in accentuating specific fluorescent polymorphic chromosome regions.


Interactions between pairs of DNA-specific fluorescent stains bound to mammalian cells. The interactions between DNA-specific fluorescence stains complexed with mitotic Chinese hamster cells were studied by spectrofluorometric and flow fluorometric techniques. The degree of binding interactions and of energy transfer between stains was determined from the intensities and shapes of fluorescence emission spectra of cells complexed with pairs of stains. The stain pairs Hoechst 33258-chromomycin A3, Hoechst 33258-ethidium bromide, and chromomycin A3-ethidium bromide exhibited efficient energy transfer from the short wavelength absorber (donor) to the long wavelength absorber (acceptor), and little competitive or cooperative binding of stains. The stain pair quinacrine-ethidium bromide exhibited both energy transfer and competitive binding. None of the stain pairs showed evidence of strong electronic interactions between stains. The magnitude of energy transfer interactions was used to estimate the quantity and distribution of the stains molecules complexed to mitotic cells. The results indicate a fairly even distribution of each of these stains along the DNA of intracellular mitotic chromosomes.


Acriflavine-Feulgen stilbene staining: a procedure for automated cervical cytology with a television based system (LEYTAS). A sample preparation and staining procedure for automated cytology with a TV based system (LEYTAS) is described. It consists of a centrifugation technique and automated acriflavine-Feulgen stilbene staining of cervical specimens. The advantages of using both the fluorescence and the absorption image of acriflavine-Feulgen stilbene stained cervical cells for a television based system are discussed.


Fluorescence background discrimination by prebleaching. A number of electrooptical techniques are described that discriminate against background fluorescence in biologic staining, whether from sample background or unbound excess stain. These techniques are based on the fluorescent decay lifetime difference between bound stain and the sample background or between the bound stain its free form. The fluorescence decay lifetimes may be measured either directly or in a combination gated photometry scheme to substantially enhance the sample background contrast. An alternative procedure uses the photochemical bleaching of fluorescent dyes under intense exposure to time discriminate with higher selectivity, sensitivity and in a more convenient fashion between diverse fluorescent molecules.


Preparation of specific antisera against adenoviruses by affinity bead immunization (ABI). Certain adenovirus types can be replicated only to low titer in tissue cultures. Other, such as adenovirus strains associated with infantile gastroenteritis, cannot be replicated in vitro. A method which allows preparation of specific antisera has therefore been evaluated. The procedure involves coupling of group-specific antibodies against adenovirus capsid subunits to CNBr-activated Sepharose 4B; reaction of crude virus suspensions with immobilized adenovirus-specific IgG; elimination of contaminating material by extensive washing using a wide pH range; and immunization with adenovirus immunogens immobilized on the beads. Efficient immunization was obtained with immunogen doses of both 50 ng and 50 microgram. The immunization procedure which has been designated affinity bead immunization (ABI) could therefore have a wide applicability in cases where the relevant immunogen constitutes a minor fraction of a crude preparation.


Amplified direct immunofluorescence (AMDI) for detection of Epstein-Barr virus nuclear antigen. Low antigen concentrations could be identified in human cells by sequential application of (a) FITC-labeled human antibodies at high dilution, (b) rabbit antiserum to the hapten fluorescein isothiocyanate (FITC), and (c) FITC-anti-rabbit globulin. The high specificity of direct immunofluorescence (a) was not affected by the amplifying steps (b) and (c). Using this AMDI technique Epstein-Barr (EB) virus nuclear antigen (EBNA) could be specifically stained with human sera up to a dilution of 1 : 4000. Owing to the high dilutions applied, unwanted antibody reactivity in the FITC-labeled serum could be blocked by preincubating with unlabeled undiluted human sera. Thus EBNA was selectively stained in EB virus producer cells. Moreover, EBNA was specifically detected in human tumor biopsy material by the use of AMDI.


Esterase staining of human peripheral blood mononuclear cells. Recently proposed esterase staining methods for the cytochemical identification of human peripheral blood monocytes and lymphocytes in our hands gave suboptimal results. Cellular purification and recommended fixation procedures appeared to decrease the esterase reactivity of leucocyte preparations. Drying for 12-18 h without further fixation of cytocentrifuge smears was found to produce  minimal loss of the staining capacity for 1-naphthyl butyrate esterase in mononuclear cells. It is hoped that the slightly modified procedure meets the need for a simple technique to define mononuclear blood cells for clinical purposes.


A partial characterization of hepatitis B e antigen. Partially purified hepatitis B e antigen (HBeAG) was prepared by ultracentrifugation, ammonium sulfate precipitation, and molecular sieve chromatography of sera obtained from asymptomatic carriers of hepatitis B surface antigen. The antigenic specificity of the HBeAG preparations was investigated further with affinity chromatography. The results indicated that HBeAG is distinct and separable from DNA polymerase activity. Columns coupled with either goat IgG prepared from antiserum to human IgG or antibody to HBeAg bound all detectable HBeAg and bound 31% and 100% of the IgG, respectively, from a partially purified HBeAg preparation. Rate zonal sucrose sedimentation and molecular sieve and ion-exchange chromatography indicated a variability in molecular weight and charge; this finding suggested a heterogenous population of immunoreactivities containing HBeAg. Our preliminary results suggest the existence of an HBeAg-IgG complex.


Ultrastructure of Bacteroides species: Bacteroides asaccharolyticus, Bacteroides fragilis, Bacteroides melaninogenicus subspecies melaninogenicus, and B. melaninogenicus subspecies intermedius. Representative strains of two subspecies of Bacteroides melaninogenicus (subspecies melaninogenicus and subspecies intermedius) and Bacteroides asaccharolyticus as well as B. asaccharolyticus strain 536B isolated from a human perirectal abscess and Bacteroides fragilis ATCC 25285 were examined by glutaraldehyde-osmium fixation, ruthenium red fixation and staining, and thorium hydroxyde staining as well as by the physical preparative techniques of critical point drying--transmission electron microscopy (CPD--TEM) and scanning electron microscopy (SEM). All strains, with the exception of B. fragilis 25285, possessed an electron-dense material external to their outer membranes. Ruthenium red staining further revealed a layer, external to the surface of the outer membrane, that was distinct for each species examined. Thorium hydroxide, as well as CPD--TEM and SEM, showed the cells to be interconnected by thin fibers that not only connected adjacent cells but also traversed several microns to connect cell aggregates.


Haemangiopericytoma in otolaryngology.

Haemangiopericytoma in otolaryngology. Haemangiopericytoma is a rare vascular tumour with variable pathological behaviour. It arises from the proliferation of special cells called 'pericytes' which are found outside the blood capillaries. There is no specific predilection for any site in the body. Clinically and histologically, it can be benign or malignant, but this differentiation sometimes becomes rather a hard task for the pathologist. Wide surgical excision is the treatment of choice. Radiotherapy and chemotherapy can be valuable adjuncts. Only fourteen cases of nasal haemangiopericytoma have been reported in the literature and these are individually set out in Table I. Another case is reported in this paper, with a review of all the tumours in the field of Otorhinolaryngology since 1942.


57Co-bleomycin kinetics in normal and tumour-bearing mice after systemic and local administration. In tumour-free and tumour-bearing mice the body clearance and organ distribution of 57Co-BLM was measured at different time intervals after iv., sc., and it. administration of the drug. No significant difference could be demonstrated in body clearance following different doses and routes of application of labelled BLM in tumour-free and tumour-bearing mice. The organ distribution studies showed higher concentrations following iv. compared to sc. or it. of 57Co-BLM; however, the activity in the ipsilateral injection sites was significantly increased after sc. and it. injection. In tumour-bearing mice the activity in the lymph nodes draining injection site was as high as that seen in the draining lymph nodes following iv. injection. However, on the contralateral side, the lymph node concentration was significantly reduced after it. injection. These results indicate on the basis or organ distribution of 57Co-BLM a rational basis for it. treatment of malignant tumours.


Increased ratio of 5 alpha-reductase: 3 alpha (beta)-hydroxysteroid dehydrogenase activities in the hyperplastic human prostate. The activities of 5 alpha-reductase and 3 alpha (beta)-hydroxysteroid dehydrogenase were assayed in homogenates of eight normal, 21 hyperplastic and four carcinomatous human prostates. Samples consisting of 300--500 microgram tissue protein in Tris buffer, pH 7.0, were incubated at 37 degrees C for 30 min in the presence of 50 nM-[3H]androgen and an NADPH-generating system started with 5 X 10(-4)M-NADP. The yield of 5 alpha- and 3 alpha-reduced metabolites, as established by using t.l.c. and g.l.c., gave an estimate of enzyme activity. The formation of metabolites denoting 5 alpha-reductase activity in normal, hyperplastic and carcinomatous tissue respectively was 28.8 +/- 47 (S.E.M.), 76.8 +/- 8.9 and 3.5 +/- 0.7 pmol 30 min-1 mg protein-1; similarly, that denoting 3 alpha (beta)-hydroxysteroid dehydrogenase activity was 69.3 +/- 6.7, 46.6 +/- 5.7 and 38.8 +/- 22.1 pmol 30 min-1 mg protein-1. In all normal prostates 5 alpha-reductase activity was lower than 3 alpha (beta)-hydroxysteroid dehydrogenase activity. Conversely, in 18 out of 21 hyperplastic prostates, 5 alpha-reductase activity was higher than 3 alpha (beta)-hydroxysteroid dehydrogenase activity. The effect of the increase in 5 alpha-reductase activity without a compensatory change in 3 alpha (beta)-hydroxysteroid dehydrogenase activity was to alter the mean ratio between 5 alpha-reductase and 3 alpha (beta)-hydroxysteriod dehydrogenase activities from 0.47 +/- 0.11 in the normal prostate to 1.84 +/- 0,19 in hyperplastic tissue. It is inferred that this change may predispose the hyperplastic prostate to asymmetrical rates of androgen metabolism and thereby contribute to the abnormal accumulation of dihydrotestosterone.


Evaluation of audiovisual teaching material in family practice: a report of review activities, 1977--1978. Audiovisual teaching materials have found increasing use in medical education in recent years, and a large number of excellent materials have been produced. The plethora of existing audiovisual teaching programs has made it difficult for educators and potential users to be aware of what is available and to select programs relevant to specific learning needs. The Audiovisual Review Committee has functioned over the last five years as a subcommittee of the Education Committee of the Society of Teachers of Family Medicine. This paper describes the experience of this group over the last two years and presents a complete listing of audiovisual teaching materials which have been reviewed and appraised during that period.


Reflection coefficients of homopore membranes: effect of molecular size and configuration. Osmotic water flow through membranes with uniform defined pores was measured for a variety of macromolecular solutes. Water flow increased linearly with applied hydrostatic pressure, allowing the effective osmotic pressure of the solutes to be estimated by extrapolation. Reflection coefficients for each solute-membrane combination were calculated and correlated with the ratio of solute size to pore size. For the same mean molecular size, proteins were found to have larger reflection coefficients than dextrans. Molecular rigidity may play a role in this difference in behavior.


Chronic relapsing experimental allergic encephalomyelitis. Correlation of circulating lymphocyte fluctuations with disease activity in suppressed and unsuppressed animals. Groups of juvenile Strain 13 guinea pigs sensitized for chronic relapsing experimental allergic encephalomyelitis (EAE) with isogeneic central nervous system (CNS) tissue in complete Freund's adjuvant (CFA) were either left to develop late-onset chronic EAE (unsuppressed), or given a series of injections of bovine myelin basic protein (MBP) in incomplete Freund's adjuvant (IFA) to suppress the disease. All unsuppressed animals developed disease and all suppressed animals remained healthy over a 27-month period of study. some unsuppressed and suppressed animals were rechallenged with CNS tissue in CFA 12 or 26 months post-inoculation (PI). Unsuppressed animals all became sick 2-4 weeks after rechallenge, while rechallenged, suppressed animals were protected, indicating that the suppression was permanent. Pathologic findings in the CNS complemented the clinical changes. Circulating lymphocyte studies were performed on animals from all groups. Early (active, high-affinity rosetting) T cell levels in unsuppressed animals showed significant decreases during exacerbations (P less than 0.01) and normal values during remissions. After rechallenge, circulating early T cells decreased in unsuppressed animals with the development of signs. In suppressed animals, early T cells showed significant elevations during, and for a short time after, the period of suppressive injections, and normal values afterwards. These levels did not change significantly after rechallenge. Late (total, 24 hour rosetting) T cell and B cell values showed minor fluctuations only which did not correlate with disease activity. These results indicate that chronic relapsing EAE can be successfully suppressed with MBP in IFA, that this suppression is permanent and that the immunologic findings presented correlate well with the clinical and pathologic facets of the disease. the findings are presented in terms of their relevance to multiple sclerosis.


Third complement component in cerebrospinal fluid in neuro-Behçet's syndrome. Conversion patterns by crossed immunoelectrophoresis. Thirteen cerebrospinal fluid specimens (CSFs) from 10 patients with neuro-Behçet's syndrome (nB) were examined for the presence of conversion products of C3 protein by means of crossed immunoelectrophoresis. C3 conversion products were found in 4 CSFs from nB patients, and the proportions of the split products to the total immunoprecipitate zones were 17, 34, 61 and 63%, respectively. CSFs with C3 conversion patterns showed higher C3 protein (beta 1C/A) levels compared with those with non-conversion, whereas the differences of C4 protein (beta 1E) levels were unremarkable. CSF with C3 conversion patterns also showed apparent inflammatory characteristics indicating blood-CSF barrier involvement which would be deduced from increased alpha 2-macroglobulin and IgA, both being regarded as sensitive internal barrier parameters: but IgA measurement was much higher than usually expected values. It is suggested that the increase in CSF C3 might be initiated by accelerated consumption of complement through the alternative pathway, leading to hypersynthetic or hyperinflux response in CSF.


Evidence for an intraaxonal transport of fixed and street rabies virus. Colchicine was used to inhibit axonal transport and to demonstrate that rabies virus spread from the peripheral inoculation site to the CNS by the retrograde axoplasmic flow. Colchicine was applied by the mean of elastomer implants around the sciatic nerve of young rats in order to obtain higher local concentrations of the drug. This procedure avoided the systemic effects of colchicine encountered with the usual treatment. To confirm the efficiency of the axoplasmic flow inhibition by colchicine, 125I-tetanus toxin was used as a marker. Uptake of colchicine by the sciatic nerve was monitored by the use of 3H-labelled colchicine. Interruption of the retrograde axoplasmic flow resulted in prevention of fixed and street rabies virus propagation. Moreover, the centrifugal spread of rabies could be inhibited using this experimental procedure.


Light and ultrastructural relationship between oxytalan fibers in the periodontal ligament of the guinea pig. The interfaces and the relationships between collagen and oxytalan fibers were observed under light and electron microscopy. Guinea pig periodontal ligament was prepared for light and electron microscopy with perfusion using Peter's buffered formalin for light microscopy and GTA-S-collidine and OSO4 for electron microscopic studies. The tissue for light microscopy was stained with a modified Gomori's aldehyde fuchsin technique, in which pre-oxidization with potassium monopersulfate was carried out before staining so as to demonstrate the oxytalan fibers. EM tissues were routinely stained with lead citrate and uranyl acetate. Two different structural relationships were observed. First, the subcomponents of the collagen and oxytalan fiber types interweave with each other; and, second, some of these two-fiber subcomponents appear attached to each other. These relationships and the known orientation of oxytalan fibers as seen in the periodontal ligament provide insight as to the function of oxytalan fibers. The oxytalan fibers may provide increased structural integrity and increased distribution of forces over a wider area of the periodontal ligament. Because of their close relationship to blood and lymph vessels in the periodontal ligament, they may also help to stabilize these elements by the same structural relationships to collagen fibers.


Lipofuscin in human tongue muscle.

Lipofuscin in human tongue muscle. In a study of 107 tongue specimens lipofuscin granules were found to be present in the muscle fibers of 93% of the cases. The pigment was not found in young individuals under the age of 18 years. The pigmented granules, which were stored in clusters of different sizes at the nuclear poles, were PAS-positive, stained black or brown with the Masson-Fontana procedure, black with Sudan black, and strong purple-pink with Ziehl-Neelson staining; yellow autofluorescene was emitted in ultraviolet light. The morphology, histochemical reactions and autofluorescence of the pigmented granules were characteristics of the endogenous lipid pigment described as lipofuscin. Statistical analyses indicated a direct correlation between increase in quantity and distribution of the pigment and increase in age (P = 0.001), in both males and females. Accumulation of the pigment in the tongue with aging was found to be similar to that in the myocardium.


Computerization of a bioassay: quantitation of slow reacting substance of anaphylaxis (SRS-A). The bioassay for SRS-A using isolated guinea-pig ileum bathed in atropine and pyrilamine has been computerized resulting in a more accurate collection and calculation of data. Areas under the polygraph tracings are calculated by a computer interfaced with the recording polygraph. After comparison of ileal responses obtained with known amounts of SRS-A standard, the computer determined the relative amounts of SRS-A in unknown samples. The program provided for a correction to adjust for changes in tissue responsiveness. Using this bioassay, we showed that ovalbumin-induced release of SRS-A from sensitized guinea-pig lung was parallel to the release of histamine measured fluorometrically. Ethanol inhibited release of these mediators of anaphylaxis in a similar fashion. Conversely, isoproterenol reduced ovalbumin-induced release of SRS-A to a greater extent than the release of histamine. The versatility of this technique should enable other bioassays to be improved in addition to greatly facilitating the determination of various pharmacologic analyses.


The blind soldier: a study dealing with the symbolism of blindness in art. An oil painting of a blind old English soldier done by James Ramsay in 1830 allows for some interesting thoughts about the use of symbolism in art and about the nature of blindness.


Estimation of prostatic size by suprapubic ultrasonography. Previous ultrasonic scanning of the prostate primarily has involved the transrectal approach. In this study the suprapubic approach was chosen because it uses basic ultrasound equipment and is atraumatic. In 29 patients undergoing prostatectomy there was a highly significant correlation (r equals 0.95) between the prostatic weight determined by this method and the postoperative weight of the adenoma.


[Trial of a synthetic absorbable suture, Polyglactin 910, in 100 urologic patients]. The authors studied the clinical tolerance of Polyglactine 910 (Vicryl) in 100 patients operated upon for urological disorders. The operations were mostly for ureterovesical reimplantations to treat reflux, pyelocaliceal kidney stones and ureteral calculi, and prostatic adenomectomy. The suture appears to be solid, and the knots do not loosen. It slides badly however and is not easy to manipulate. When used to close cavity walls this solid suture did not cause a higher percentage of abscess than catgut, when used to suture the urinary excretory tract the authors observed, in one case only, multiple calcification around the ureterovesical suture points and those used to close the bladder. The suture is not suitable for vaginal repair operations during the surgical treatment of incontinence. The authors conclude by defining the properties of a suture they would like to see become available: one which has the same solidity, is easier to manipulate, slides more easily, and, above all, has a different texture: a monofilament rather than a braided suture.


Serum beta-2-microglobulin levels in urological cancer. Serum beta-2-microglobulin levels were measured in patients with renal, vesical and prostatic cancer. Measurements were made only on samples with a serum creatinine less than or equal to 105 mumol./l. to eliminate the possibility of elevated beta-2-microglobulin being a result of impaired renal function. This criterion eliminated 28 to 50 per cent of the patients with bladder cancer and 73 per cent of those who had undergone nephrectomy for renal carcinoma, which, obviously, limits the value of beta-2-microglobulin measurement for the surveillance in these cancers. Beta-2-microglobulin values in patients with prostatic cancer were seldom increased to more than 3.0 mg./l. In bladder cancer patients with normal serum creatinine the frequency of an elevated serum beta-2-microglobulin increased with the increase in tumor stage.


The results of prostatectomy: a symptomatic and urodynamic analysis of 152 patients. There were 152 patients with prostatism investigated by inflow cystometry and pressure flow analysis of micturition before and after elective prostatectomy. Many symptoms attributed to obstruction were found to be owing to bladder instability, which was demonstrated in 60 per cent of the patients. The symptomatic improvement postoperatively was accompanied by a decrease in the incidence of instability to 25 per cent. The repeat urodynamic studies and symptom analysis demonstrated a surgical success rate of 88 per cent.


Resource for managing sleep disorders.

Resource for managing sleep disorders. To enhance physicians' knowledge of the prevalent problem of sleep disorders, a 50-minute videotape program designed as a learning and self-assessment experience in evaluating and treating these disorders was coproduced with the Network for Continuing Medical Education. The program has been presented nationally to about 25,000 medical professionals. The mean correct score was 50% for the pretest, 69% for the instructional section, and 79% for the posttest; the composite score on the posttest was 59% higher than that on the pretest. Physicians' responses indicated strengths and weaknesses in their knowledge of sleep disorders, thus delineating future directions for medical education in insomnia, narcolepsy/cataplexy, nightmares, night terrors, sleepwalking, and enuresis. All of these sleep disorders can be best evaluated and treated by the general physician.


[Treatment with NK 631 (new bleomycin analog) of oral and maxillofacial cancer (author's transl)]. Eight cases of malignant tumors of the head and neck were treated with NK 631 on a dosage schedule of 10 mg at a time 3 times weekly, by intravenous one-shot injection or intravenous drip infusion, to observe its therapeutic effects and adverse reactions. The treatment was assessed markedly effective in 3, moderately effective in 1 and ineffective in 4 of them. The treatment was also assessed moderately or markedly effective in 3 and ineffective in 2 out of squamous cell carcinoma cases. Hematologic findings, serum electrolytes and enzymologic findings were normal, but the pulmonary function examinations revealed a tendency for PaO2 to decrease slightly. In 1 case where frequent cough was observed, the cough was mitigated on withdrawal of the treatment. The adverse reactions that evolved included fever, alopecia, eruptions, nausea and vomiting, and pigmentation of the nail. To summarize these findings, the authors were impressed with NK 631 and that the agent would exert an excellent antitumor effect, compared with bleomycin, and that its effect would evolve at the early stage of its treatment. Fixed drug eruption was observed as an adverse reaction of this drug in 1 case; however, the adverse reactions of this bleomycin analog appear similar to those of its parent compound, bleomycin.


[Toxicological studies on pepleomycin sulfate (NK631). VI. Chronic toxicity of pepleomycin in dogs (author's transl)]. Chronic toxicity and its recovery of pepleomycin sulfate was studied in both sexes of beagle dogs. At dose levels of 0.3, 0.15 and 0.075 mg/kg, pepleomycin was administered intramuscularly to dogs for 180 successive days. Two dogs of the 0.15 mg/kg dose group were used for recovery test for 35 days. As general findings, the decrease of food intake, the loss of body weight, ulceration of foot pad, nail root necrosis and onychoptosis, ulcer of tongue and labia, and alopecia, dermatitis and necrosis at friction sites were observed more severely in the 0.3 mg/kg dose group of both sexes, especially in male, than those in bleomycin were. In the dose groups of 0.15 and 0.075 mg/kg, their findings were observed as slightly as those in bleomycin were. The death occurred in the 0.3 mg/kg dose group of both sexes. The lesions of liver and kidney were recognized in the 0.3 mg/kg dose group of both sexes, severely in male, on histopathological findings. Additionally severe fibrosis of lung was observed in one of the 0.3 mg/kg dose group of female. In general chronic toxicity of pepleomycin was revealed more severely in the 0.3 mg/kg dose group than that of bleomycin was, but in the dose groups of 0.15 and 0.075 mg/kg difference between their toxicities was not significant. In addition, chronic toxicity of pepleomycin in dogs showed more severely in male and its recovery was hardly recognized during its period. The maximum safety dose in this studies was estimated to be between 0.075 and 0.15 mg/kg in dogs.


[Clinical experience with NK 631 in malignant tumors of skin (author's transl)]. NK 631, a new derivative of bleomycin, was clinically used in 14 cases of malignant tumors of skin. The results obtained were remarkably effective in 7 cases, effective in 3 cases, slightly effective in 2 cases and ineffective in 2 cases. The effective ratio was 71% when calculated regarding slightly effective as ineffective. Compared with bleomycin, we were impressed that the side effects observed with NK 631 were similar to those with bleomycin, but the influences for lung were smaller and slighter than the latter.


Human acute myelogenous leukemia antigens defined by simian antisera: evidence for leukemia-associated antigens distinct from immune response-associated alloantigens. Leukemic blasts from a patient with acute myelogenous leukemia (AML) and peripheral blood T- and B-lymphocyte subpopulations from his genetically identical normal twin were analyzed with the use of the simian antiserum-defining AML antigens and a rabbit antiserum to immune response-associated (la)-like antigens. Blast cells from the patient consistently reacted with both reagents, whereas the B-lymphocyte populations from the patient's normal identical twin reacted only with the rabbit anti-la serum and in no instances reacted with the antiserum to AML cell antigens. Blast cells from the AML patient significantly stimulated the lymphocytes of his normal twin and his own remission leukocytes, whereas the cells from the normal twin failed to stimulate the cells of the patient. These results suggested the existence on AML cells of tumor-associated antigens that are distinct from various other well-characterized normal human alloantigens and differentiation antigens including B-cell antigens. Changes were reported in the expression of leukemia-associated antigens and Ia-like antigens on the cells of an AML patient undergoing chemotherapy as well as in the ability of the simian antisera to distinguish antigens specific for myeloid leukemias from lymphocytic types of leukemias.


Human tumor-lymphocyte interaction in vitro. VI. Specificity of primary and secondary autologous lymphocyte-mediated cytotoxicity. A T-cell-enriched lymphocyte subset of samples from 15 tumor patients was tested for primary cytotoxicity against autologous tumor cell preparations and against 1-3 different allogeneic tumor cell preparations from biopsy material. Allogeneic cytotoxicity occurred in only 1 of 10 patients with autologous reactivity. The lymphocytes of 14 patients were cultured with autologous cells from biopsy material for 6 days. These lymphocytes killed autologous targets, but only 1 patient's lymphocytes were cytotoxic against 1 of the 4 allogeneic tumors tested. Cocultivation with allogeneic cells from biopsy specimens generated cytotoxicity toward the sensitizing allogeneic cells in 3 of 9 test combinations. In 2 of 3 instances the effectors were also active against the autologous tumor cells. Cytotoxicity in primary and secondary tests occurred thus only rarely against allogeneic targets. This indicated either the presence of individual tumor-related antigens on the cells from biopsy material or reflected the histocompatibility restriction of T-cell-mediated cytotoxicity.


Effect of Propionibacterium acnes on the cellular immune responses to tumor-specific antigens on malignant rat liver cells. The effect of Propionibacterium acnes on the cellular immune responses to tumor-specific membrane antigens was investigated by microcytotoxicity assays (MA) and 51Cr release assays (CRA) with use of mesenteric lymph node cells (LNC) of syngeneic BD IV and BD VI rats. BD rat liver cell lines transformed in vitro by chemical carcinogens were used as target cells with tumor-specific antigens. By MA, the LNC from rats that were inoculated with malignant liver cells under the adjuvant effect of heat-killed P. acnes showed significant cytotoxic response to the target cells but not to nonmalignant liver cells. By CRA, these LNC did not show specific cytolysis to the malignant liver cells. Assays with various target cells derived from BD rat liver and inhibition tests with syngeneic and xenogeneic antisera against tumor-specific antigens on the malignant liver cells proved that LNC reacted with tumor-specific individual or tumor-specific cross-reacting antigens on the malignant liver cells. Cytotoxic responses against the malignant liver cells were not demonstrated even by MA with use of the LNC from rats inoculated with either the malignant liver cells or P. acnes alone. LNC from the rats inoculated with both nonmalignant liver cells and P. acnes were not cytotoxic to malignant or nonmalignant liver cell lines.


[Disorders of heart rhythm during bicycle ergometry in ischemic heart disease]. During a test on a bicycle ergometer disorders of cardiac rhythm were revealed in 41 (20.5%) out of 240 patients with ischemic heart disease who underwent coronary arteriography. It is concluded that disorders of cardiac rhythm are mostly encountered in patients with ischemic heart disease attended by stenosing changes of the coronary arteries and disorders of left ventricular contractile function as demonstrated by ventriculography. In a few cases ventricular extrasystole may be the only sign of myocardial ischemia in patients with ischemic heart disease.


[Healing of argon laser lesions of the rabbit iris (author's transl)]. Healing of perforating and low dose argon laser lesions of pigmented rabbit irides was studied by light microscopy. Low dose coagulation produced brown stained areas with fibrinous exudation of the surrounding tissue. Healing proceeded with little infiltration of leucocytes leading to an atrophic iris scar with numerous pigment macrophages. Laser iridotomies were closed within 28 days by a scar, shaped by proliferating cells and newly formed collagen fibrils. Lesions of the epithelium of the lens were demonstrated by hemalum staining of the nuclei, even in cases, when no lens opacities could be detected by biomicroscopy.


Immunologic features of a carcinogen-induced murine bladder cancer: in vivo and in vitro studies. Certain in vivo and in vitro immunologic features of carcinogen-induced murine bladder cancer have been studied. The consistency of tumor induction, its natural history, and immunogenicity both in vivo and in vitro render this syngeneic murine bladder tumor a suitable model for immunologic studies. Pre-immunization of strain C3H/Hen mice with mid-gestational fetal cells did not protect the animals from tumor challenge. Sera of mice immunized with mid-gestational fetal cells were not cytotoxic to cultured tumor cells in a microcytotoxicity assay indicative of dissimilarity between the tumor associated antigen and the syngeneic mid-gestational fetal antigen.


Fluorescence histochemical and ultrastructural observations on preterminal noradrenergic axons following ligation or treatment with 6-hydroxydopamine. This study was designed to compare the effects of 6-hydroxydopamine (6-OHDA) treatment with those of mechanical constriction of noradrenergic preterminal fibres in the rat. Constriction produced a progressive accumulation of glyoxylic acid induced fluorescence, attributable to noradrenaline, in nerve trunks proximal to applied ligatures. The electron microscope showed that this accumulation comprised large numbers of granular vesicles in swollen axons. Degenerative changes were also present and the axonal microtubules were distorted or absent. Preterminal fibres also contained accumulated noradrenaline histofluorescence after 6-ODHA (100 mg/kg) treatment, though this accumulation was not as focal as it was after constriction. The ultrastructural appearance of preterminals from 6-OHDA-treated rats closesely resembled that seen proximal to constrictions. Swollen axons were present. These contained many granular vesicles and exhibited degenerative changes. The microtubules were disrupted or absent. It is suggested that 6-OHDA produces accumulation of noradrenaline storage vesicles and other organelles in preterminal fibres because the primary lesion of the terminals presents a barrier to intra-axonal transport of these organelles.


Are pancreatic beta-cells under vagal control? To elucidate the importance of cholinergic innervation for pancreatic beta-cells in vivo, atropine sulphate (0.2 mg/kg body weight) was subcutaneously injected into mice 4 times a day for 10 days. Control animals received 0.9% NaCl. Paraffin sections of the pancreas were stained for beta-cells with aldehyde fuchsin and for alpha-cells (glucagon cells) with silver according to Grimelius. Endocrine and exocrine cell nuclei were measured with an ocular screw micrometer. The light absorbance at 550 nm wave-length of aldehyde-fuchsin-stained islet sections was measured with a microscope photometer. Atropine caused no loss of body weight or apparent food consumption. The nuclei of beta-cells shrank significantly in response to atropine; A550 of islet surfaces showed a significant negative correlation to beta-cell nuclear size. Acinar cell nuclei near islets also became smaller after atropine treatment but to a lesser extent. No such change was observed in the alpha-cells. A trophic influence of the vagal nerve may be important for the long-term control of beta-cell function.


Coronary-artery bypass surgery in stable angina pectoris: Survival at two years. European Coronary Surgery Study Group. 768 men aged under 65 with angina pectoris, at least 50% obstruction in two or more major vessels, and a left-ventricular ejection fraction greater than or equal to 0.5 took part in a prospective randomised trial of the effect of coronary-artery bypass on prognosis. 373 patients were alloted to medical and 395 to surgical treatment. There was no significant difference between the two groups in the distribution of variables recorded at the time of randomisation. 1. "surgical" patient was lost to follow-up. 26 "surgical" patients did not undergo surgery and 50 "medical" patients were operated on. All these 76 patients were retained in their original treatment groups for the analysis. At 2 years there was no significant difference in mortality between the two groups. A significant difference was, however, found in the subset of patients with three-vessel disease, survival being significantly better for surgical patients. Operative (in-hospital) mortality was 3.6% in all operated patients and 1.5% in the last third. On average, 1.9 grafts per patient were inserted in the two-vessel-disease subgroup and 2.4 grafts per patient in the three-vessel-disease subgroup. Graft-patency rate was 90% within 9 months and 77% between 9 and 18 months after surgery. Symptomatic improvement was significantly better and deterioration less in the surgical group.


Comparison between systemic and oral antimicrobial prophylaxis in colorectal surgery. In a prospective randomised trial in which 93 patients undergoing elective colorectal operations were given a short prophylactic course of metronidazole and kanamycin orally or systemically, postoperative sepsis occurred in only 3 (6.5%) of those given antimicrobials systemically, compared with 17 (36%) of those given oral prophylaxis (P less than 0.01). 15 of the 17 infections in patients who received antimicrobials orally were due to kanamycin-resistant bacteria present in the colon at operation. Bacterial overgrowth of Staphylococcus aureus was recorded in 6 of the patients who received oral therapy. Antibiotic-associated pseudomembranous colitis occurred in 7 patients, 6 of whom had received prophylaxis orally. These results indicate that oral administration of prophylactic antimicrobials in colon surgery should be avoided because of the risks of bacterial resistance, superinfection, and antibiotic-associated pseudomembranous colitis. Systemic per-operative antimicrobial prophylaxis is safer and more effective.


The loin-pain/haematuria syndrome.

The loin-pain/haematuria syndrome. Nine patients with the loin-pain/haematuria syndrome are described. The previously reported clinical features of severe recurrent loin pain and tenderness in young women, the relationship of symptoms to use of oral contraceptives, and the demonstration of intrarenal vascular abnormalities by angiography are confirmed. Histology revealed minor non-specific abnormalities, and electron microscopy showed fibrin in the afferent arteriole and in glomerular capillaries of one patient only. No diagnostic changes were found on renography. In all patients except one the heparin-thrombin clotting-time was abnormal, suggesting increased platelet activity or release and providing further evidence of a vascular disorder.


Are apolipoproteins better discriminators than lipids for atherosclerosis? Plasma-levels of major lipids (cholesterol, triglyceride, high-density-lipoprotein cholesterol), two major apolipoproteins (apo-B and apo-A1), and two ratios (total-cholesterol/apo-B and apo-A1/apo-B) were studied in 218 survivors of myocardial infarction and 160 controls. Apolipoproteins were as good as lipids as discriminators between the populations under the age of 50 and better in the sixth to eighth decades.. Furthermore, values of total-cholesterol/apo-B and apo-A1/apo-B obtained from controls and normolipaemic survivors of myocardial infarction gave a bimodal distribution. The protein moiety of lipoproteins is a better discriminator than lipids between atherosclerotic subjects and controls.


Liver disease among homosexual males.

Liver disease among homosexual males. 5% of 2612 homosexual males attending genitourinary clinics were found to be hepatitis-B surface-antigen (HBsAg) positive. Liver biopsy was done in 25 who had abnormal liver-function tests but no symptoms or signs of liver disease, and 14 (56%) of these proved to have chronic active hepatitis or active cirrhosis. Neither the liver-function tests nor the viral markers in serum reflected the severity of the liver disease. 38% of a group of 118 HBsAg positive patients were HBeAg positive, and sexual contacts of these individuals may be at serious risk of infection.


Obstructive nephropathy: successful evaluation with radionuclides. A non-invasive test of nephron function, the renal parenchymal transit time (P.T.T.) index, is shown to be as accurate as antegrade pressure measurements in diagnosis of renal outflow obstruction. It is more sensitive than conventional methods of demonstrating the associated obstructive nephropathy. P.T.T. is useful in following the effects of surgery, or of temporising, in patients with confirmed or potential outflow tract obstruction. Renal radionuclide P.T.T. analysis gives the functional information essential for the management of patients in whom intravenous urography reveals a dilated renal pelvis.


When should pre-term babies be sent home from neonatal units? 20 randomly selected infants of 33 weeks gestation and under at birth were allowed to go home from the neonatal unit provided they were clinically well and had passed the nadir of their postnatal  weight-loss and provided home conditions were satisfactory. Weight-gain at home was satisfactory and there was no increased rate of hospital readmission compared with 20 randomly selected pre-term infants who were discharged at a more traditional weight of 2200 g. There is therefore little justification for the widespread practice in Britain of delaying discharge of preterm infants until they reach a predetermined weight (usually 2000--2500 g).


Depression in women after perinatal death. Post-partum depression was studied with the Beck inventory during a 6-month period in two groups of over 300 women. The difference in prevalence of depression between women who had had live births and women whose babies were stillborn or died in the first 7 days of life was influenced by age. At 6 months, post-partum depression was just as common in women aged under 24 whose babies had survived as in women of the same age whose babies had died.


Uromucoids and urinary stone formation.

Uromucoids and urinary stone formation. Crystal formation was studied in fresh urine samples after rapid concentration to standard osmolarity in a rotary evaporator at 37 degrees C. Uromucoids promoted calcium oxalate and calcium phosphate crystal formation and also induced clustering of calcium phosphate precipitates. It is postulated that uromucoid precipitation is the first stage in stone formation. Minerals then deposit on the uromucoid precipitate and the whole complex becomes attached to the renal tubules.


Prostacyclin: a solution to some problems of extracorporeal circulation. Experiments in greyhounds. In cardiopulmonary-bypass experiments in greyhounds the effects of adding prostacyclin, prostacyclin plus heparin, and heparin alone to the extracorporeal circulation were compared. With heparin alone platelet count and function were reduced, the pressure differential across the arterial filters rose, platelet deposits were found on the arterial filters, and plasma-fibrinogen levels fell. Plasma from these dogs was toxic to fetal-mouse hearts in culture. With prostacyclin alone, fibrinogen levels fell, but the platelets were preserved. With a combination of prostacyclin and heparin, platelet count and function were maintained, there was no consumption of fibrinogen, and there was little deposition on the arterial filters.


Cimetidine and gastric cancer.

Cimetidine and gastric cancer. Three patients with dyspeptic symptoms who were being treated with the H2-receptor blocking drug, cimetidine, were later found to have gastric carcinoma. It was not possible to determine whether the association was fortuitous, whether the drug had masked the neoplastic change, or whether it was involved in some other way. Repeated clinical and endoscopic evaluation is essential in patients on this treatment for any length of time.


Controlled comparison of day-hospital and outpatient treatment for neurotic disorders. 106 patients with anxiety, phobic, and depressive neuroses referred to the outpatient clinics of 6 psychiatrists were randomly assigned to outpatient care or two forms of psychiatric day care. Ratings of symptoms and social adjustment were recorded before treatment and after 4 and 8 months in 89 patients. There were no important differences in the outcome of day care and outpatient treatment although patients were more satisfied with the outpatient service. Because outpatient care is more economical of time and personnel it should be preferred to psychiatric day care for many neurotic disorders.


Factors associated with cardiac mortality in developed countries with particular reference to the consumption of wine. Deaths from ischaemic heart-disease in 18 developed countries are not strongly associated with health-service factors such as doctor and nurse density. There is a negative association with gross national product per capita and a positive but inconsistent association with saturated and monounsaturated fat intake. The principal finding is a strong and specific negative association between ischaemic heart-disease deaths and alcohol consumption. This is shown to be wholly attributable to wine consumption.


Percutaneous transluminal angioplasty for severe renovascular hypertension due to renal-artery medial fibroplasia. Percutaneous transluminal renal angioplasty (P.T.R.A.) was performed in a 43-year-old woman with medial fibroplasia of the right main renal artery that was causing high-grade stenoses and severe hypertension. The procedure led to a virtually complete dilatation of the stenosing lesions and a prompt and lasting fall in blood-pressure to normal. Hypersecretion of renin from the right kidney was also promptly suppressed. The procedure was well tolerated by the patient and no complications were discerned. P.T.R.A. deserves further investigation to assess its role in the management of renovascular hypertension.


Oestrogen receptors and prognosis in early breast cancer. In a study of the role of oestrogen-receptor analysis in early breast cancer the oestrogen-receptor content of the tumour was estimated in 286 patients undergoing mastectomy. These patients were followed for up to 39 months, and the recurrence of disease was noted in relation to the presence or absence of oestrogen receptor.Recurrence-rates were significantly higher in patients whose tumours did not contain receptors than in those whose tumours did. This same relationship was seen when women with and without axillary metastases were considered separately. The highest rates of recurrence were in women with axillary lymph-node involvement whose tumours lacked oestrogen receptors. Women without axillary-node involvement whose tumours lacked oestrogen receptors showed the same high rate of recurrence as all women with axillary-node involvement. The oestrogen-receptor content of a primary breast cancer appears to be an independent guide to early recurrence of the disease.


Chlorpropamide alcohol flushing and diabetic retinopathy. "Mason-type" diabetics (mild diabetes which is dominantly inherited) are relatively free of retinopathy. Alcohol almost invariably causes facial flushing in these patients when they are given chlorpropamide (chlorpropamide alcohol flush, C.P.A.F.). 291 non-insulin-dependent diabetics were examined to see whether there was a difference in frequency of retinopathy between C.P.A.F. positive and negative cases who were of comparable age and duration of diabetes. Retinopathy was commoner and often severe in CPAF negative patients. Blindness from retinopathy was almost confined to C.P.A.F.-negative cases. Lens opacities, on the other hand, were equally common in both groups. Since C.P.A.F. is an inherited trait, retinopathy in non-insulin-dependent diabetics is to a considerable extent, although not entirely, determined by genetic factors.


High incidence of undetected neoplasia in maldescended testes. Testicular biopsy specimens from 4 (8%) of 50 men previously treated for maldescended testes had a carcinoma-in-situ pattern. 2 of these men had adjacent invasive carcinoma (seminoma and embryonal carcinoma). The patient with embryonal carcinoma had an enlarged testis but the 3 other patients with neoplasia had no clinical signs or symptoms of malignancy. Routine follow-up including testicular biopsy in young men with maldescended testes is essential because of the increased risk of malignancy and may detect testicular neoplasia at a stage when orchidectomy alone is curative.


Comparison of effects of different combined oral-contraceptive formulations on carbohydrate and lipid metabolism. Oral glucose-tolerance tests were performed and fasting serum cholesterol and triglyceride levels measured in 1628 Caucasian women taking combined oestrogen/progestagen oral contraceptives (o.c.) and 577 women not taking O.C. The former were divided into six groups according to the composition of the O.C Glucose tolerance deteriorated in all O.C. groups containing oestrane progestagens (nortestosterone-derived) or the gonane, norgestrel, but was unaltered by O.C. containing a pregnane progestagen (derived from progesterone). The greatest deterioration was with O.C. containing 75 microgram or more oestrogen, and this was associated with impairment of the early insulin response to glucose. In O.C. containing a pregnane progestagen insulin secretion was unaffected. In the remaining O.C. groups insulin secretion was increased; this was most pronounced with the O.C. containing a gonane progestagen. Serum-cholesterol was elevated only with O.C. containing 75 microgram or more oestrogen and an oestrane progestagen and tended to be lower in O.C. containing a gonane progestagen. O.C.-induced hypertriglyceridaemia was oestrogen-dose-related, and this effect was potentiated by the pregnane progestagen. The gonane progestagen antagonised oestrogen-induced hypertriglyceridaemia.


Unrecognised nocturnal hypoglycaemia in insulin-treated diabetics. Overnight metabolic studies in 39 poorly controlled insulin-treated diabetic patients aged 9 to 66 years showed hypoglycaemia (blood-glucose less than 2 mmol/1) in 22 patients; it lasted 3 h or more in 17. Hypoglycaemic symptoms were very mild or absent, but 19 patients had other features of overtreatment with insulin. These included lethargy, depression, night sweats, morning headaches, fits (3 patients), glycogen-laden hepatomegaly (3), and acquired tolerance to high doses of insulin (mean 1 u/kg/24 h). The best clinical clue to recurrent nocturnal hypoglycaemia was the intermittent occurrence of symptoms, however "mild" and infrequent these appeared to be. Reduction of insulin by a mean of 25% in these patients (without change of species) did not result in loss of overall control; 1 patient with recurrent ketoacidosis was stablished on 40% of his initial dose. It is difficult, sometimes impossible, to achieve good overnight control with conventional once or twice daily insulin therapy. Since patients readily become tolerant of low blood-glucose levels, reliance on urine tests and symptoms of hypoglycaemia as a guide to dosage easily produces a spiral of overtreatment.


Biopsy evidence of left ventricular myocardial abnormality in patients with mitral-leaflet prolapse and chest pain. Left ventricular endomyocardial biopsy specimens were obtained at cardiac catheterisation in 11 patients with mitral-leaflet prolapse, chest pain, and normal coronary arteries. Specimens were examined for birefringence before and after contraction induced by adenosine triphosphate and were also subjected to histochemical analysis. Biopsy specimens from a control group of patients with rigorously defined normal left ventricular function and another control group with poor left ventricular function were also studied. Biopsy specimens from patients with mitral-leaflet prolapse showed a markedly subnormal birefringence response to adenosine triphosphate, with the group mean value intermediate between values for controls with normal and poor left ventricular function. Histochemical studies in the prolapse group also revealed an abnormal pattern, particularly for monoamine-oxidase activity. This is further evidence of a cardiomyopathy in this symptomatic subgroup of patients with mitral-leaflet prolapse.


Cause of anaemia in malaria.

Cause of anaemia in malaria. 51Cr studies showed that after complete eradication of malarial parasites erythrocyte life-span is reduced for 4-5 weeks and that the reduction is associated with the presence of complement-containing immune complexes on the red-cell surface. The rate of erythrophagocytosis in the spleen is increased during this period. 59Fe studies of erythropoiesis indicated a mild degree of marrow depression. A very important factor causing anaemia in malaria appears to be a complement-mediated immune process.


The puzzle of "urethral syndrome": a possible answer? 82 strains of slow-growing, CO2-dependent, gram-positive organisms have been isolated from the urine of 9 male and 73 female patients. 100% of the men and 93% of the women had urinary symptoms at the time of isolation, and 66% of the specimens showed pyuria. These organisms might account for the urinary symptoms of some patients previously diagnosed as having "urethral syndrome". Preliminary data show that, if appropriate antibacterial treatment is given, most of these patients become symptom-free, and the organism can no longer be isolated from the urine.


Endoscopic choledochoduodenostomy for common bileduct obstructions. Endoscopic choledochoduodenostomy (E.C.D.S.) was done in six patients with common bileduct obstructions. After E.C.D.S., jaundice disappeared and the patient's general condition improved. There were no complications. E.C.D.S. may be a suitable alternative to conventional surgery in patients with common bileduct obstructions.


Simplifying radiological examinations. The urogram as a model. The diagnostic yield of one and three film urograms was compared with that of complete examinations to determine whether a moderately complex examination could be simplified without loss of important diagnostic information. Although sensitivity was high (88-93%) and was not altered by increasing the complexity of the examination, the definitive disease diagnoses were more accurate with the three film rather than the one film studies. Specificity increased from 69% to 77-80% with the more complex examinations. A strategy based on terminating the examination if the single film urogram is normal with a three-film examination in positive cases might effect considerable savings, both economic and in terms of gonadal radiation dose, without serious diagnostic loss.


"Have you eaten laburnum?".

"Have you eaten laburnum?". In an average summer over three thousand children are admitted to hospital in England and Wales because of laburnum poisoning. It is suggested that laburnum is not as dangerous as has been thought and that many of these admissions are unnecessary.


Epidemic hepatitis B caused by commercial human immunoglobulin. An epidemic of acute hepatitis B followed the administration of human immunoglobulin to members of the staff of a mission hospital in India and their families. Jaundice developed in 123 (38%) of 325 persons inoculated. Hepatitis-B surface antigen was detected in three of the batches of immunoglobulin which were available for testing.


Delayed separation of the umbilical cord, widespread infections, and defective neutrophil mobility. In six infants, from two families. the umbilical cords were still attached at 3 weeks of age. Five of these developed severe local and disseminated infections from which four died. Two of these children were tested, and both, including the survivor, had defective neutrophil mobility; in the survivor this was improved in vitro and in vivo by ascorbic acid. It is suggested that a primary genetic defect of a contractile protein could explain the association. The sixth child, with delayed cord separation but normal neutrophil mobility and no excess of infections, who has survived without special treatments, also has mastocytosis, apparently inherited independently.


One-hour blood-xylose test in diagnosis of cow's milk protein intolerance. In this prospective study, 18 consecutive children suspected of cow's milk protein intolerance (C.M.P.I.) on clinical grounds were investigated before and after challenge with cow's milk. One-hour blood-xylose, serum-IgE, eosinophil count, serum-complement (C3) and a jejunal biopsy specimen for histology and disaccharidase activity were obtained from all patients before challenge. Serum-complement was measured again 24 h after the beginning of the challenge and the first three tests were repeated on the fifth day. A second jejunal biopsy was obtained in 9 of the first 10 children. 15 of the 18 children were considered to have C.M.P.I. The one-hour blood-xylose test emerged as a simple and valuable test in the diagnosis of C.M.P.I. A significant drop (51--81%) in the one-hour blood-xylose level was observed in all 15 patients. In all patients but one it dropped below 25 mg/dl. Mean value (+/- S.D.) for the group was 47.5 +/- 11.1 mg/dl before and 17.2 +/- 4.2 mg/dl after cow's milk ingestion.


Progression of diabetic nephropathy.

Progression of diabetic nephropathy. Although deterioration of renal function in diabetic nephropathy varies considerably from one diabetic to another, its rate is constant in individuals. For each patient there is a linear relation between period (months) which elapses from the time serum creatinine becomes greater than 200 mumol/l and the inverse the inverse of the serum-creatinine. The observation is of practical importance in predicting the time at which end-stage renal failure will develop, so that treatment can be planned in advance.


Prenatal diagnosis of congenital adrenal hyperplasia (21-hydroxylase deficiency) by HLA typing. Congenital adrenal hyperplasia (C.A.H.) due to 21-hydroxylase deficiency is an HLA-linked recessive disorder. HLA-A and B antigens are expressed on amniotic cells. Prenatal diagnosis of C.A.H. by HLA typing of families and amniotic cells was attempted in two at-risk families. In one family HLA typing indicated that the fetus would have C.A.H., and this prediction was confirmed after birth. In the second family, HLA typing indicated that the fetus would be an unaffected, phenotypically normal carrier of the disease gene, and this prediction was also confirmed after birth.


Jejunal morphology in multiple sclerosis. In a prospective survey jejunal biopsy samples from 11 patients with multiple sclerosis on normal diets were compared with those from controls matched for age and sex. Quantitative histology, morphometry, and electron microscopy showed no difference between the two groups. Since there was no evidence that gluten sensitivity plays a role in the aetiology of multiple sclerosis, the wide adoption of gluten exclusion by patients seems unjustified.


Severe hypertension after ingestion of an appetite suppressant (phenylpropanolamine) with indomethacin. Severe systemic hypertension developed in a patient who took indomethacin shortly after ingesting an appetite suppressant ('Trimolets') containing phenylpropanolamine. The hypertension was attributed to a drug interaction whereby the inhibition of prostaglandin synthesis by indomethacin exacerbated the sympathomimetic effects of phenylpropanolamine. It is suggested that withdrawal of preparations containing phenylpropranolamine from general use should be considered in view of their potential for adverse reactions with other commonly used drugs and their doubtful therapeutic value.


Successful therapy of advanced arteriosclerosis obliterans with prostacyclin. Five patients with advanced arteriosclerosis obliterans of the lower extremities, as evidenced by resting pain, ischaemic ulcers, and focal necrosis, received intra-arterial infusions of prostacyclin at doses of 5--10 ng/kg/min for 72 h. Within 2 days of termination of the infusion, pain at rest had disappeared in all patients. In three of the five, the necrosis had completely regressed and the ischaemic ulcers healed within 2 months. The other two patients showed considerable improvement. Prostacyclin therapy was not associated with changes in the radiographic appearance of the major blood-vessels. However, muscle blood-flow, as measured by xenon-133 clearance, was significantly increased both during prostacyclin infusion and for the 6 weeks of measurement after its termination.


Disorders of chloriuretic hormone secretion. Experimental evidence supports the existence of a circulating substance, natriuretic hormone, which augments electrolyte excretion. Because such a hormone probably acts by inhibiting chloride reabsorption in the thick, ascending limb of the loop of Henle it would more accurately be called chloriuretic hormone. Chloriuretic hormone must have an action which resembles that of loop diuretics such as frusemide and ethacrynic acid. An excess of chloriuretic hormone could explain all the manifestations of Bartter's syndrome, whereas a deficiency could account for Gordon's syndrome. Hyporeninaemic hypoaldosteronism may develop in subjects who are unable to increase chloriuretic hormone concentrations appropriately in response to progressive impairment of renal function.


Effect of different operation policies on mortality from bleeding peptic ulcer. The outcome in bleeding gastric and duodenal ulcer has been compared in two hospitals in Nottingham which take their patients randomly from a common source according to a fixed daily admission rota. Bleeding seemed equally severe in the two groups but 66 (32%) of 206 patients were operated upon at hospital A compared with 44 (46%) of 96 at hospital B (P=0.03), where operations also tended to be done earlier. Postoperative mortality-rates were virtually identical at the two hospitals, but at hospital B the overall death-rate from bleeding ulcer was higher, partly because of its higher operation-rate. The value of an aggressive surgical policy in bleeding peptic ulcer is questionable.


Tests of prolactin secretion in diagnosis of prolactinomas. Prolactin-secreting tumours of the pituitary were identified and treated by transsphenoidal microsurgery in fourteen infertile females with hyperprolactinaemia. Resting prolactin levels were 590--9000 mU/1 (mean 3400). In seven patients, tomography of the pituitary fossa was normal and resting prolactin levels were 590-6000 mU/1 (mean 3400). In these patients the pre-operative diagnosis prolactinoma in these patients was made by demonstrating loss of the normal circadian prolactin profile and impaired prolactin response to intravenous thyrotrophin-releasing hormone (T.R.H.) and metoclopramide stimulation. Prolactin response to the acute oral administration of L-dopa and bromocriptine was of less diagnostic value. Preoperative assessment of anterior pituitary function identified abnormalities other than hyperprolactinaemia in four patients (28%). Post-operative assessment indicated that microsurgery was curative in twelve patients (86%), selective in all, and without significant side-effect. It is concluded that dynamic tests such as T.R.H. and metoclopramide stimulation have considerable value in identifying hyperprolactinaemic patients with prolactin-secreting adenomas, particularly those which are radiologically occult.


Specific allergic sensitsation to filarial antigens in tropical eosinophilia syndrome. Reaginic antibodies to antigens from the human filarial parasites Wuchereria bancrofti and Brugia malayi and the animal parasite Dirofilaria immitis were studied by histamine release from basophils in 7 patients with tropical eosinophilia (T.E.) and 18 patients with other manifestations of filarial infection (lymphatic changes or symptomless microfilaraemia). All the patients had antibodies to all three filariae but T.E. patients were more highly sensitised. T.E. patients responded more to antigens from microfilariae than did patients with non-T.E. filariasis and responded more to microfilarial antigens from the human parasites than to those from the animal parasite. These findings support the view that T.E. is a form of occult filariasis which results from host hypersensitivity to the microfilarial stage of parasites which, in other individuals, cause the more common lymphatic manifestations of filarial disease.


Low frequency of chlamydial antibodies in patients with Crohn's disease and ulcerative colitis. Serum samples from 55 patients with Crohn's disease and from 23 patients with ulcerative colitis were tested for antibodies to Chlamydia trachomatis immunotypes by a micro-immunofluorescence technique. Antibody titres of 1:8 or greater against several immunotypes were detected in 14.5% of patients with Crohn's disease and in 21.7% of those with ulcerative colitis. These figures resemble the incidence in a healthy, non-venereal-disease population. Furthermore, there was no correlation between the presence of antibody and such factors as duration of symptoms, localisation of disease, or disease activity. These findings indicate that there is no reason to believe that Crohn's disease involves chlamydiae or that examination for chlamydial antibody is helpful in diagnosis.


Cellular origins of the fetal-haemoglobin-containing cells of normal adults. The origin of the small population of adult red cells which contain Hb F (F-cells) has been studied in a clonal disorder of haemopoiesis, polycythaemia rubra vera (P.R.V.). In eleven patients who had not received cytotoxic therapy F-cells comprised less than 0.1% to 11.9% of the circulating red cells, compared with 0.34% to 4.6% in 21 haematologically normal controls. Two additional patients were glucose-6-phosphate-dehydrogenase heterozygotes in whom the clonal nature of the P.R.V. could be demonstrated directly; they had F-cell values of 2.1% and 8.3%. These observations indicate that F-cells arise from the same population of stem cells as other adult cells and not from a separate stem-cell pool.


Classes of antibodies in acute gonorrhoea. In patients with acute gonorrhoea the antibody responses to envelope antigens of Neisseria gonorrhoea were investigated with a sensitive enzyme-linked immunosorbent assay. IgG antibodies predominated even early in the disease but IgM and IgA antibodies were present. Surprisingly there were circulating antibodies linked to secretory piece. These were largely but not entirely IgA and were presumed to have been absorbed from sites of local production.


Tuberculosis in immunosuppressed patients. 11 patients who developed tuberculosis while on long-term immunosuppressive-drug therapy are described. The indications for immunosuppressive therapy were varied, but all patients received high doses of corticosteroids with azathioprine in addition in 2 cases and chlorambucil in a third. The diagnosis was delayed in all cases because of suppression of symptoms, so that 4 patients died, 3 directly as a result of tuberculosis. 3 of the remaining patients were critically ill at the time of diagnosis but recovered with antituberculosis chemotherapy; whereas the other 4, although not critically ill, had advanced cavitated pulmonary tuberculosis with miliary spread in one case. A clear policy of investigation and, if indicated, chemoprophylaxis is necessary for all patients in whom long-term immunosuppressive therapy, especially with prednisolone, is contemplated. A high index of suspicion for tuberculosis must be maintained with regular clinical and radiological review.


An outbreak of gastroenteritis in young children caused by adenoviruses. During October and November, 1978, gastroenteritis developed in 17 of 24 young children aged between eight months and two years from an R.A.F. station in the U.K. The illness, in which diarrhoea was always the predominant symptom, had an incubation period of eight to ten days and lasted about a week. It seemed to be transmitted from child to child, and in all but one instance parents and older siblings remained well. Stool specimens from 14 of the affected children were examined bacteriologically and virologically, and a highly significant association was found between the presence of adenovirus particles in stools, identified by electron microscopy, and the acute stage of the illness. This evidence suggests that an adenovirus was the cause of this outbreak of gastroenteritis.


Increased shelf-life and feulgen staining intensity of a modified trichloracetic acid-Schiff reagent. Two modified formulae for the preparation of trichloracetic acid (TCA)-Schiff reagent have been presented in this communication. The modifications involve addition of 250 mg or 500 mg of trichloracetic acid per 100 ml of a 0.5% aqueous solution of basic fuchsin in place of NTCA which means 1.63 g of TCA per 10 ml of distilled water to be added to 100 ml of the dye solution. The use of TCA-Schiff reagents prepared by the modified method reveals a considerably faster Feulgen staining, a more intense colouration of the nuclei and a longer shelf-life of the dye-SO2 reagents as compared with the standard TCA-Schiff of Bloch and Godman. Possible interpretations for these phenomena have been presented.


Hoffman's violet and dahlia as specific stains for animal chromosomes. The paper deals with staining of the chromosomes of animal testicular materials with two basic dyes, Hoffman's violet and dahlia of the triphenylmethane group, following iodine-dye procedure. The important finding, as presented herein, is that iodinated alcohol after staining can be substituted with various acids, both organic as well as inorganic, all of which act as trapping agent preventing leaching of the dye that binds with the chromosomal DNA. It is clear from this study that RNA is not involved by this process of staining, since treatment of stained sections with cold phosphoric acid at 5 degrees C for 20--25 min and then stained also reveals perfect colouration of the chromosomes without any cytoplasmic staining. The in vitro absorption properties of Hoffman's violet have also been presented herein. The probable mechanism of action of these dyes has been suggested.


A differential staining technique for vertebrate histology. A staining method is described for studying micro-anatomy of different vertebrate tissues in the light microscope. A staining sequence of celestin blue--erythrosin--orange G--fast green with mordanting in phosphomolybdic acid yields a satisfactory differentiation and fine colour contrast in various tissues. The efficacy of the method was tested on different avian and mammalian tissues.


Simian rotavirus (SA 11) in serodiagnosis of human rotavirus infections. Tests were made on 169 sera from children up to 10 years of age for rotavirus antibodies by indirect immunofluorescence and complement fixation tests. The simian strain SA 11 served as a substitute antigen for the human rotavirus. Furthermore, the SA 11 complement-fixing antigen was compared with a commercial antigen of the bovine rotavirus strain NCDV. It was demonstrated that SA 11 which can be more easily propagated than most bovine strains, may be used as a substitute antigen in complement fixation and indirect immunofluorescence for serodiagnosis of human rotavirus infections.


Bleomycin in combination chemotherapy of metastatic Wilms tumor: a late recurrent case, refractory to conventional therapy. A patient with a late recurrence (13 years) of Wilms tumor is presented. After relapse with conventional chemotherapy, an objective partial remission was obtained with bleomycin, vinblastine, cyclophosphamide, and cis-platinum. Stable disease was noted when bleomycin was withheld. Bleomycin in this combination appeared to effect a good response, and further studies of chemotherapy of Wilms tumor should include bleomycin in combination with other active agents.


Bleomycin induces cataract formation in newborn rats in association with modifications of crystallin synthesis. The antitumor antibiotic Bleomycin induces the formation of cataract when injected into newborn rats but not when injected into rats after 12 days or older. The development of these lesions is strictly associated with progressive biochemical modifications of the crystallins as demonstrated by electrophoresis on one and two-dimensional gels. When the lenses of the rats are observed histologically, the first modifications appear between 7 and 13 days after the injection of the drug concomitantly with the first biochemical modifications. Later on the fiber cells start to degenerate and there is a gradual destruction of the whole center of the lens.


[Reverse transcription of the RNA of eukaryotes and viruses. Conditions for obtaining a long product]. The receiving of full-size DNA-copies of RNAs is necessary for molecular hybridisation experiments as well as for synthesis of recombinant bacterial plasmids with eucariotic DNA sequences. Some authors received such cDNAs for different RNAs with a help of variations in reaction conditions. In this article it is shown that such empirically chosen conditions mainly had an influence on a secondary structure of RNA-templates and that only in such a case the synthesis of the DNA-product with sizes corresponding to the RNA-template is possible.


[Conformational state and mechanism of functioning of gramicidin A]. Theoretical conformational analysis of the antibiotic gramicidin A HCO--L-Val--Gly--L-Ala--D-leu--L-Ala--D-Val--L-Val--D-Val--(L-Trp--D-Leu)3--L-Trp--NHCH2CH2OH has been carried out by stagewise computations of a serie of LD penta-decapeptide analogs, which approximated the structure of the natural compound at the final stage. The potential surface of the LD-peptide skeleton of the gramicidin molecule is shown to predetermine the existence of a set of pi4LD--Pi6LD structures. Low-energy helical structures with no hydrogen bonds have also been revealed, which are due to compensational relations between hydrogen bonding and nonbonded energies. Inclusion of D-Val into the amino acid sequence discriminate against alpha-helix, while Trp and Leu residues contribute to a formation of pi4LD and pi6LD helices and to a reduction of energy differences between them. Conformational properties and geometrical parameters of the lowest-energy helical structures of gramicidin provide transport of protones and of all alkali metal ions. A mechanism of cation transportation through the gramicidin trans-membrane channel is discussed.


[Circular dichroism of DNA complexes with dyes. III. Effect of latent optical activity and the structure of the complexes]. Circular dichroism anisotropy was studied both theoretically and experimentally for the complexes of DNA and dsRNA with dyes (proflavine, 2,7-di-t-butyl proflavine, "Hoechst-33258") and antibiotics (distamycin A, netropsin and olivomycin). Theoretical analysis showed that general features of CD anisotropy, revealed in the previous studies (CD components--delta epsilon parallel to and delta epsilon perpendicular--are ten times or more bigger than the CD-effect without orientation, and delta epsilon parallel to approximately 2 delta epsilon perpendicular) are due to the existence of a specific effect named "latent" optical activity (LOA). This effect can be observed in many cases of non-chiral symmetrical chromophores if they are oriented. The effect is due to the excitation of an electrical dipole transition and a perpendicular magnetic dipole transition (or quadrupole transition) of a molecule. The amplitude and the sign of the LOA-effect depends on the orientation of the chromophores with respect to the light beam; with a random orientation the mutual compensation of LOA-effects of different chromophores happens and no LOA-effect appears. The analitycal expressions relating the value of LOA-effect of the system with electronical characteristics of the chromophores and the geometrical parametra of their arrangement was obtained. The experimental data obtained for the oriented complexes of DNA and dsRNA with proflavine made it possible to determine an angle between the chromophore and the plane perpendicular to the DNA axis--gamma. For the calf thymus DNA gamma = = + 1.8 +/- 0.4 degrees, for the phage T2 DNA gamma = + 2.2 +/- 0.4 degrees, and for phage f2 dsRNA gamma=--3.5 +/- +/- 0.5 degrees. These results, obtained at relatively low concentrations of the bound proflavine (r approximately 0.01), are in accordance with the intercalating mode of the dye binding. A study of CD anisotropy of DNA complexes with other ligands showed that many different chromophores possess LOA-effect. This phenomenon can be used to obtain both spectroscopic and structural information about the systems similar to those reported here.


["Revertase" project: results of carrying out the scientific program]. The article summarizes the data obtained within the scope of the "Revertase" project organized in 1972 under the guidance of Scientific Councel on Molecular Biology. The scientific program included four main topics: synthesis of complementary DNA via reverse transcription on the various RNA templates and their use as tools in molecular biology, enzymatic synthesis of structural genes in double DNA-polymerase system and their propagation by means of genetic engineering, enzymology of reverse transcription, problems of molecular oncornavirology. It is stressed that the success in realization of this wide and intensive research program was achieved due to cooperative efforts and division of labour between scientists of different specialities due to international cooperation within the frame of the project. Appearance of more than 80 publications between 1973 and 1977 clearly manifested that the program was fullfilled and led to creation of a new area of research absent in the given countries before functioning of the "Revertase" project.


[Electron microscopic study of the DNA products of reverse transcription]. DNA-products synthesized on pre-mRNA's from rat liver and rabbit erythroidal bone marrow cells, on rabbit globin mRNA and the RNA-templates themselves have been examined by electron microscopy. In spreading conditions providing extension of molecules the sizes of DNA-products corresponded to sizes of RNA templates. Globin mRNA, pre-mRNA as well as cDNA synthesized on these templates in the presence of actinomycine D were seen as single-stranded molecules by electron microscopy. The preparations of cDNA synthesized in the absence of actinomycine D on the pre-mRNA template were represented by double-stranded molecules along side with sigle-sranded. Up to 10% of double-stranded DNA-product appeared to be branched; all branches of such molecules had the thickness and rigidity of double-stranded structures. The possible ways of formation of branched structures are discussed.


Detection of small numbers of monoclonal B lymphocytes in the blood of patients with lymphoma. The definition of complete remission and early relapse in patients with lymphoid malignancy is complicated by the difficulty of recognizing the presence of small numbers of malignant lymphocytes in a population of normal lymphocytes. We have used a method based on the cytofluorometric detection of lymphocytes having homogeneous amounts of surface immunoglobulin of one light-chain class to study blood from patients with lymphomas. The test is capable of reliably detecting 10 per cent or less of monoclonal B lymphocytes in normal blood, and it shows a high incidnece (30 to 40 per cent) of previously unsuspected monoclonal B lymphocytes in patients who were though to have no abnormal blood cells according to standard morphologic technics. The ability to estimate the number of such cells may facilitate the planning of therapy and make more meaningful the concept of complete remission.


Specificities of antibodies to acetylcholine receptors. Antisera to acetylcholine receptor purified from the electric organs of Torpedo and Electrophorus were highly species specific, but showed limited cross-reaction with receptors from all mammalian species tested. Antisera to receptor purified from rat muscle cross-reacted extensively with receptor from the muscles of other mammals. Antibodies to receptor from Electrophorus cross-reacting with receptor from Torpedo were affinity purified. Subsequently these showed increased cross-reaction with receptor from mammalian muscle. Sera from patients with myasthenia gravis reacted best with receptor from human muscle, but such sera also cross-reacted with receptor from other mammals, especially squirrel monkeys. These sera cross-reacted slightly with receptor from Torpedo, but not detectably with receptor from Electrophorus,


Myasthenia gravis: review of diagnosis and management. Although the cause of myasthenia gravis is still unknown, its pathogenesis appears clear: immunologic attack on synaptic receptors in muscle causes receptor deficiency, decreased miniature endplate potentials, and decrements in the compound action potentials evoked from muscles on repetitive stimulation of peripheral nerves. In addition to the involvement of skeletal muscle, some MG patients may manifest subtle alterations of the function of heart, lung, smooth muscle, and CNS, indicating that this is truly a systemic disorder. Modern therapy involves adjusting treatment to the needs of individual patients. Anticholinesterases, calcium, ephedrine, potassium, and germine partially correct the defect in neuromuscular transmission; prednisone, ACTH, cytotoxic drugs, antilymphocyte serums, gamma globulin, thoracic duct drainage, plasmapheresis, and thymectomy partially modify the abnormalities of the immune system.


[Use of psychotropic substances with antidepressive action in pain control]. Starting with the conviction that chronic oncological pain needs an interdisciplinary approach, the use of antidepressant tricyclic compounds in association with other techiques in antalgic therapy is commented on. In spite of the possible side effects, it is concluded that the drugs are definitely effective, and that in addition to combating the depressive component, when present, their analgesic action potentiates the effect of other drugs.


Endometrioid carcinoma of the ovary. A clinicopathologic, histochemical, and electron microscopic study. A material consisting of 23 endometrioid ovarian carcinomas, including 2 rare endometrioid tumors with argyrophil cells, was analyzed clinically and with the use of various histochemical staining methods. Electron microscopy was performed in 3 cases. Apical neutral mucin mixed with sulfate and carboxyl groups was typical for the cells at the light microscopic level, and a prominent Golgi complex with many small secretory vesicles was commonly seen at the ultrastructural level. The nuclei had concentric nuclear bodies and nucleoli with mesh-basket appearance. There was a positive correlation between the grade of differentiation and clinical stage. The corrected 5-year survival rate was 80% in stage I, 50% in stage II, 17% in stage III, and 0% in stage IV; the overall 5-year survival rate was 46%. Signs of increased endometrial estrogen activity were found in 6 of the 12 postmenopausal patients. In the whole series, 6 endometrial carcinomas were found simultaneously with the ovarian carcinoma.


The development of preschool children of heroin-addicted mothers: a controlled study. Disturbances of growth and behavior in infants and toddlers of women addicted to heroin during pregnancy have been reported in uncontrolled studies. In this study, 3- to 6-year-old children of heroin-addicted mothers were compared to three other groups matched for age, race, sex, birth weight, and socioeconomic status. Heroin-exposed children weighed less and were shorter than those in the comparison groups; 14% had a head circumference below the third percentile. Heroin-exposed children were rated by parents as less well adjusted than control children and they differed significantly in perceptual measures and on subtests of the Illinois Test of Psycholinguistic Abilities and McCarthy Scales of Children's Abilities relating to the process of organization. These findings suggest that chronic intrauterine exposure to heroin may affect growth and behavior as well as perceptual and learning processes in preschool children.


Apparent allosterism by avian myeloblastosis virus reverse transcriptase and E. coli DNA polymerase I. A recent report (1) presented evidence for allosterism in reverse transcription by Mason-Pfizer monkey virus reverse transcriptase and by E. coli DNA polymerase I. Our experiments also demonstrate these apparent cooperative effects when synthesis is catalyzed by either avian myeloblastosis virus DNA polymerase, feline sarcoma virus DNA polymerase, or E. coli DNA polymerase I (large fragment). We show that the apparent cooperativity depends on the use of oligo(dT)12-18 as primer. However, if the polymerase reaction products are isolated chromatographically, then the polymerases obey classical Michaelis-Menten kinetics with respect to substrate and enzyme concentrations. These results suggest that the cooperative effects are an acid precipitation artifact. The results are also consistent with the enzyme operating by a distributive mechanism with the oligo(dT)12-18 primer.


Isolation of human alpha-fetoprotein in two fractionation steps and demonstration of homogeneity. Human alpha-fetoprotein (hAFP) has been isolated from cord serum in 40% yield using an isolation procedure consisting of only two major steps: affinity chromatography followed by preparative polyacrylamide gel electrophoresis (PAGE). The final product appeared homogeneous on the basis of five independent criteria for purity. Sodium dodecyl sulfate gel electrophoresis (SDS-PAGE) demonstrated a single polypeptide chain with molecular weight of 71,000. The protein exhibited an apparent isoelectric point (pI') of 4.85, molecular radius of 3.0 nm and a valence (net H+/molecule) of 21.9 derived from computation of analytical PAGE data. The two-step isolation procedure made it possible for a single operator to isolate milligram amounts of hAFP in a matter of weeks.


Genome of avian myelocytomatosis virus MC29: analysis by heteroduplex mapping. The virion RNA of avian myelocytoma virus MC29 was hybridized to full genome length DNA of the Prague strain of Rous sarcoma virus and analyzed by heteroduplex mapping in the electron microscopy. The results show that MC29 specific sequences for which there are no homologous counterparts in the Rous sarcoma virus genome make up a contiguous stretch of RNA about 1.8 kilobases long. These sequences are located approximately in the middle of the genome, replacing the 3' half of the gag gene, the entire pol gene, and the 5' portion of the env gene, which are absent from MC29. This MC29 specific genetic substitution may contain information for the leukemogenic transformation of the host cell.


Antigenic drift in type A influenza virus: peptide mapping and antigenic analysis of A/PR/8/34 (HON1) variants selected with monoclonal antibodies. Variants of A/PR/8/34 (HON1) influenza virus, having hemagglutinin molecules with probably a single altered antigenic determinant, were isolated by growing the virus in the presence of the monoclonal hybridoma antibody PEG-1. The variants were analyzed by peptide mapping and characterized antigenically by using PEG-1 and four other monoclonal hybridoma antibodies to PR8 hemagglutinin. Peptide maps of the large hemagglutinin polypeptide, HA1, from 8 out of 10 variants showed a single changed peptide. This peptide from two of the variants was analyzed, and in each case a serine residue in the wild-type hemagglutinin was replaced by leucine in the variant. Although these eight variants showed identical peptide maps, one could be discriminated antigenically from the others with one of the hybridomas. (The peptide maps represented about one-third of the HA1 molecule.) Of the other two variants, one gave the same HA1 map as the wild type, but could be distinguished antigenically from wild-type virus by two of the hybridomas. The other was unique, and could be distinguished, both antigenically and by peptide mapping, from the other variants. Since a large number of the variants selected with PEG-1 showed the same peptide change, it is likely that this alteration in amino acid sequence (serine to leucine) was responsible for the inability of the variants to bind PEG-1 monoclonal antibody. We do not know, however, whether the changed amino acids were located within the antigenic sites or whether the change occurred somewhere else in the hemagglutinin molecule and altered the determinants through conformational changes.


In vitro transformation of mouse bone marrow cells by the polycythemic strain of Friend leukemia virus. Strains of Friend leukemia virus (FLV) that are associated with polycythemia contain the defective spleen focus-forming virus (SFFV). To determine whether the transforming ability of FLV was affected by the presence of this second agent, DBA/2J mouse bone marrow cells were infected in vitro. Criteria for transformation were the establishment of permanent lines, growth on semisolid agarose, and the production of tumors at the site of inoculation in syngeneic hosts. Two lines of immature hematopoietic cells that grow in suspension originated from the infected cultures. Each has an almost diploid karyotype (38-39 chromosomes) and 3-4 metacentric chromosomes. These transformed cells express gp71 viral envelope glycoprotein and p30 viral core protein antigens. Virus production was measured by reverse transcriptase (RNA-dependent DNA polymerase) activity of the virions released into the medium. The virus, assayed in vivo for infectivity, has SFFV activity but is attenuated for leukemogenicity. The stimulation of hemoglobin synthesis in the cells grown in medium supplemented with dimethyl sulfoxide or hexamethylene bisacetamide indicates that the cells are erythroid in origin. SFFV may have a function analogous to erythropoietin in influencing the process of transformation by FLV.


The attenuation of delta 9-tetrahydrocannabinol and morphine of the quasi-morphine withdrawal syndrome in rats. The effect of delta 9-tetrahydrocannabinol (THC), morphine, haloperidol and chlordiazepoxide on the exhibition of the signs of the quasi-morphine withdrawal syndrome was studied in rats. In preliminary studies approximately equi-sedative doses of these drugs were chosen. Morphine and THC produced a very similar degree of suppression of the signs of the quasi-morphine withdrawal, but unlike morphine, the effects of THC were not reversed by the narcotic antagonist, naloxone. The dopamine receptor antagonist, haloperidol, produced a moderate suppression of the withdrawal syndrome and chlordiazepoxide was without significant effect. It is concluded that THC is of very similar potency to morphine in suppressing the quasi-morphine withdrawal syndrome, but its activity in this regard does not appear to be dependent upon the availability of opiate or dopamine receptors, nor is it due to sedation alone.


[A new method of cytological staining].

[A new method of cytological staining]. The haematological method (Giemsa, Pappenheim, Wright, ecc.) is renowed for karyological legible, while the cytologic method (Papanicolau, Shorr, haematoxylon-eosine, etc.) for its view of the whole. Antithesis between two methods was subject polemics and discussions after put in round table promoted by Acta cytological in 1958. The present work suggests for the attention of cytologists an attempt at technical reconciliation of two methods, it obtained with a new coloration method that uses like fixative a cadmiumalcoholacetic mixture, like nuclear colouring the basic fuchsine maturing in presence of formic aldehyde and acetic acid, like cytoplasmatic colouring a triacid orange-eosine-blue mixture of aniline.


Central nervous system lesions biopsied or treated by CT-guided needle placement. Computed tomography has been utilized to guide a needle into central nervous system lesions as small as one centimeter in diameter. Direct puncture has drained many abscesses effectively, provided material for immediate Gram stain and subsequent culture, and permitted irrigation with appropriate antibioties, Multilocular cerebral lesions and loculated extracerebral fluid collections have been aspirated and treated with this technique. Aspiration of known cystic tumors has reduced their mass and obviated some palliative decompressions. Unresectable gliomas and metastases have been biopsied prior to chemotherapy or radiotherapy. Hemorrhage and other rare complications of biopsy may be detected and immediately treated.