Effect of interferon on mouse cells chronically infected with murine leukaemia virus: kinetic studies on virus production and virus RNA synthesis. NIH/3T3 cells chronically infected with the Moloney strain of murine leukaemia virus were incubated with interferon (IF). There was no effect on virus production during the first 4 h, but thereafter an antiviral state gradually developed, reaching a maximum at about 12 h. When IF was removed, the antiviral state (expressed in terms of inhibition of release of virus) remained constant for 10 h, after which there was an abrupt return to the normal rate of virus release. Analysis of IF-treated cells showed that there was a three to fourfold increase in the amount of virus RNA in the nucleus at 48 h after IF addition, and still a slight increase at 72 h. There were no increases in the amounts of virus RNA in the cytoplasm during 72 h after the addition of IF. These results agree with the postulate that IF inhibits a late stage in the maturation of virus in chronically infected cells.


Isolation and purification of a polymeric form of the glycoprotein of rabies virus. Of the three major proteins associated with the rabies virus membrane, only the glycoprotein was found to be located on the external surface of the virus membrane. Glycoprotein prepared by treatment of rabies virus with Triton X-100 and purified by isoelectric focusing was found to be homogeneous with respect to size and isoelectric point. This material, which is free of phospholipids, is able to protect in vaccination experiments against a lethal challenge infection with rabies virus. The apparent mol. wt. of this component isolated under non-denaturing conditions is approx. 400000. The same material analysed by SDS polyacrylamide gel electrophoresis (PAGE) was found to consist solely of polypeptide chains of the G protein (mol. wt. 80000). A minor glycoprotein (gp 50), detected by PAGE of the Triton X-100 released material, appeared to be a breakdown product of the G-protein. Therefore the detergent released material represents homopolymers of the G-protein. Whether the antigenic determinants reside on the monomeric subunit or are a property of the polymeric form of the G-protein is discussed.


A direct plaque assay for hog cholera virus. Direct plaque formation with representative strains of hog cholera virus (HCV) has been obtained using several pig kidney cell lines under agar overlay. HCV-infected cells appear as hazy plaques when viewed against an indirect light source, and as white plaques after neutral red staining. HCV assay by direct plaque procedure is rapid and convenient and gives infectivity titres identical to the fluorescent focus assay technique.


Virus-like particles associated with the mitochondria of ethidium bromide treated transformed cells. A cell line resistant to ethidium bromide has been developed from baby hamster kidney cells transformed by Rous sarcoma virus. The parental cells are normally non-producers but the resistant cell line appears to produce virus-like particles that are associated with the mitochondria as visualized by electron microscopy and determined biochemically.


Studies on Cotia virus--an unclassified poxvirus. This paper is a report of studies on Cotia virus; this had been first isolated in 1965 in Brazil and was subsequently shown to be a poxvirus. Cotia virus grew in a wide range of cell cultures and on the chick chorioallantois (CAM), Its growth characteristics are similar to those of other poxviruses. Microscopy showed virus factories or type B inclusions appearing before infectious progeny virus could be demonstrated. Type A inclusions appeared later, after development of progeny virus; these were shown by electron microscopy to differ from the type A inclusions of cowpox and other poxviruses and they have been termed Cotia bodies. Immunofluorescent staining also showed ring structures which appeared before the development of Cotia bodies. The growth of Cotia virus in human embryo lung (HEL) cells was sensitive to inhibitors of DNA and protein synthesis but was resistant to a concentration of rifampicin which inhibited vaccinia virus. Sharing of antigens between the Cotia virus and vaccinia virus was shown by gel precipitation tests and immunofluorescent staining. There was no cross neutralization between Cotia virus and vaccinia virus nor did anti-Cotia sera neutralize representatives of other poxvirus groups.


Further evidence for deletion of envelope glycoprotein (gp69/71) sequences in formation of Moloney-murine sarcoma virus. Moloney-murine sarcoma virus (S+L- strain of M-MSV) has been nonproductively cloned in murine and non-murine host cells (S+L- cells) and the expression of Moloney leukaemia virus (M-MuLV) 30000 mol. wt. core protein (p30) and envelope glycoprotein (gp69/71) were studied by radioimmunoassay. Antigenic determinants of the M-MuLV p30 were associated with the sarcoma virus genome in these non-productively transformed cell clones studied, while the determinants of M-MuLV gp69/71 were not. The absence of envelope-associated glycoprotein expression in sarcoma virus transformed cells was confirmation of biological studies demonstrating that rescued sarcoma virions acquire envelope-associated properties of host range, neutralization and interference from rescuing helper virus, and further evidence that the M-MuLV gp69/71 sequences have been deleted during the formation of the M-MSV. During the course of these studies, it was also found that S+L- dog cells were releasing into culture supernatant large amounts of the p30 antigenic determinant, apparently as a soluble antigen.


Anoxic block and recovery of axoplasmic transport and electrical excitability of nerve. Axoplasmic transport of cat sciatic nerves was studied in vitro in a chamber in which maximal alpha action potentials could also be elicited. After initiation of N2 anoxia, electrical responses fell to zero at an average time of 22 min. A shorter time to zero of 11 min was seen during a second period of anoxia. A good recovery of both action potential responses and axoplasmic transport occurs after a period of anoxia lasting 1--1.5 hr. An apparent failure of recovery of axoplasmic transport was seen after 2 hr of anoxia with a good recovery of electrical responses. Axoplasmic transport tended to return toward normal when more time was allowed for recovery after anoxia. An adequate supply of approximately P was shown to be present by measurement of ATP and creatine phosphate levels. The delay in recovery of transport thus signifies a failure of utilization of approximately P by the transport mechanism. Longer periods of anoxia and recovery were limited in vitro and for this reason, ischemic anoxia was produced in vivo. Blood pressure cuffs were placed on the upper thigh of cats and maintained for times of 1--8 hr at pressures of 300-310 mm Hg. Then, recovery times up to 7 days were allowed. It was shown that axoplasmic transport could gradually recovery after an anoxia lasting up to 6-7 hr if sufficient recovery times were allowed. A possible explanation for the delay in the recovery of axoplasmic transport and the disassociation in the earlier recovery of electrical responses as against the recovery of transport was discussed.


Gliotoxin analogues as inhibitors of reverse transcriptase. 1. Effect of lipophilicity. The reaction scheme, developed for the synthesis of the gliotoxin analogue 2, was found to be of general applicability for analogues with varying substituents at N(1) and C(2). Analogues 11b-g prepared by this method are inhibitors of reverse transcriptase (RNA-directed DNA polymerase). Their inhibitory activity seems to be related to the lipophilicity of the effector molecules: the most lipophilic compound is the most active inhibitor. The techniques of reversed-phase thin-layer chromatography with silylated, precoated plates as well as reversed-phase high-performance liquid chromatography were used to measure the relative lipophilicities; both techniques gave analogous results.


Gliotoxin analogues as inhibitors of reverse transcriptase. 2. Resolution and X-ray crystal structure determination. A novel, simple, and efficient method for the chemical resolution of epidithiodioxopiperazines is reported, which is based upon covalent formation of diastereomers. This method might be a general one for the resolution of chiral cyclic disulfides. Dithiol 5, prepared from 2 by reduction with NaBH4, was allowed to react with the disulfenyl chloride 8 to yield 9 and 10, which were separated by short-column chromatography on silica gel. From these, the optically pure enantiomers 11 and 12, respectively, were obtained by reduction with NaBH4, followed by reoxidation with I2-pyridine. In this way the precursor 7 of the resolving agent could also be recovered. The absolute configurations of 11 and 12 were derived from CD spectra. Kinetic asymmetric transformation of the gliotoxin analogue 2 with the diphosphine 6 gave a 19% enrichment in one enantiomer of the starting material. Surprisingly, both enantiomers were found to inhibit reverse transcriptase, the RNA-dependent DNA polymerase, to the same degree, indicating that there is no relation between this property of epidithiodioxopiperazines and their bridgehead configurations. From the X-ray crystal structure determination it can be seen that there is a considerable torsional and conformational strain in compound 2, which might enhance the ease of cleavage of the S-S bond. A possible relationship between this property and the biological activity of 2 is discussed.


Underutilization and misuse of audiovisual modes of communication by senior faculty. Guest lectures by 10 chairmen of medical school departments, five professors, and five associate professors were videotaped. Their means of communication were than analyzed in detail. The faculty did best when using the written word for lecture synopses. The majority misused or underutilized audiovisual methods at least part of the time. A number of basic mistakes were made during slide presentations, including intermixing of illegible slides with very clear ones. All of the speakers projected still photos (slides) but relied very little on movie films and videotapes. The sampling reported here implies that audience preferences and needs are not being met well by some faculty members, who may benefit from workshops or other training in audiovisual modes of communications.


Antibody-linked cytotoxic agents in the treatment of cancer: current status and future prospects. Antibodies against tumor cell surface antigens have been used as selective carriers of anticancer drugs, which themselves lack selectivity. Although such antibodies have been demonstrated in tumor hosts, xenogeneic antitumor sera should provide larger yields of better-defined antitumor antibodies for therapeutic purposes. This review examined factors that influence the immune response to tumor-associated transplantation antigens (TATA) and the methods for rendering tumor cells more immunogenic. Consideration was also given to techniques for elimination of irrelevant immunoglobulin molecules. These could involve purification of both antitumor sera and TATA fractions for immunization, as well as tailoring of the immunization protocol. Various toxic agents that have been linked to antitumor globulins with retention of agent and antibody activity were tabulated: alkylating drugs, antibiotics, antimetabolites, cell surface agents, protein synthesis inhibitors, and unconventional anticancer agents that selectively convert nontoxic arsenicals or halides into cytocidal derivatives. The methods by which effective conjugates can be produced and their possible mode of action were described for the different types of agents. Several problems inherent in this modality of tumor therapy include: 1) the necessity of binding therapeutically effective amounts of antitumor agent, 2) ensuring of delivery of drug in active form to target sites, 3) avoidance of host reactions to foreign proteins, and 4) possible emergence of resistant tumor cell populations. Antibody-linked cytotoxic agents may find their greatest use in the eradication of small numbers of circulating tumor cells and micrometastases remaining after removal of primary tumors.


Immunologic methods for the identification of cell types. II. Expression of normal mouse mammary epithelial cell antigens in mammary neoplasia. Mouse mammary epithelial (MME) cell antigens were cell type-specific and were retained, to a large extent, by MME cells after neoplastic transformation. These MME cell antigens were expressed in mammary tumors of BALB/c, C3H, GRS/A, RIII, and Is/Bi mice tested and were not expressed in tumors whose normal counterpart was other than mammary tissue. They were not dependent on cell culture conditions or on the presence of murine mammary tumor virus; therefore, they can be used in an ubiquitous cell type marker for MME cells in both normal and neoplastic tissues.


In vivo modulation of thymus-leukemia antigens on mouse leukemia cells and thymocytes: retention of modulating antibody on the cell surface. Inoculation of RADA1, ASL1, and ERLD murine leukemia cells into the peritoneal cavities of (C57BL/6J x A/TL--)F1 mice hyperimmunized against thymus-leukemia (TL) cell-surface antigens rendered most cells insensitive to lysis in vitro by guinea pig complement even in the presence of TL antiserum. Thymocytes of A/J mice were similarly modulated by passive injection of TL antiserum. In all cases, retention of some modulating antibody on the surfaces of most cells modulated in vivo for 1--27 days was indicated by: 1) acquisition of sensitivity of modulated cells to lysis by absorbed rabbit complement; 2) positive immunofluorescence reactions for mouse IgG on the surfaces of modulated cells; and 3) release of cytolytically active TL antibody from cells into the circulation of unimmunized mice following transfer of modulated cells. Reversal of modulation of RADA1 cells was complete in some experiments within 24 hours after transfer to unimmunized mice, by which time all indications of cell-bound TL antibody were lost. These results indicate that even long-term modulation of TL antigenicity in vivo does not result in a complete loss of modulating antibody (presumably attached to TL antigens) from the cell surface.


Metabolism of concanavalin A-affinity molecular variants of alpha-fetoprotein in rats. Serum half-lives of unfractionated rat alpha-fetoprotein (AFP), concanavalin A (Con A)-nonreactive AFP, and Con A-reactive AFP in normal 45- to 55-day-old male and female F344 rats were determined to be 19 hours. No significant differences were seen between the half-lives of unfractionated AFP and either of the two Con A-affinity molecular variants of AFP studied. No evidence of interconversion of the Con A-affinity AFP variants during 24 hours in the circulation was found. These results supported the conclusion that the proportions of Con A-affinity molecular variants of AFP in the sera of fetal, newborn, and hematoma-bearing rats result from differences in the relative synthesis rates of the two variants.


Secondary in vitro generation of cytolytic T-lymphocytes (CTL's) in the murine sarcoma virus system. Virus-specific CTL induction across the H-2 barrier. Following in vitro stimulation of murine sarcoma virus Moloney isolate (M-MuSV)-immune spleen cells with syngeneic antigenically related Moloney leukemia cells, highly efficient cytotoxic T-lymphocytes (CTL's) were generated. The cytotoxic effect was directed only against H-2-compatible target cells bearing M-MuSV tumor-associated antigens (TAA). However, in a cold target competition assay a weak but detectable capacity to block CTL activity was also obtained when allogeneic Moloney leukemia cells were added. Moreover, when M-MuSV-immune spleen cells from mice inoculated with virus 14 days previously were stimulated by allogeneic Moloney leukemia cells, a strong cytotoxic effect toward syngeneic and allogeneic tumor cells bearing M-MuSV TAA was elicited.


Type-common CP-1 antigen of herpes simplex virus is associated with a 59,000-molecular-weight envelope glycoprotein. The CP-1 antigen of herpes simplex virus type 1 (HSV-1) is a glycoprotein found in the soluble portion of infected cells, in detergent extracts of infected cell membranes, and in the envelope of purified virus. Antisera were prepared against a further purified form of CP-1 prepared from HSV soluble antigen mix; a glycoprotein, gp52, isolated from detergent-treated infected cells; and detergent extracts of purified virus. Each of the antisera reacted with CP-1 to give a single immunoprecipitin band of identity, and each antiserum neutralized the infectivity of HSV-1 and HSV-2. Our results suggested that the type-common determinants involved in the stimulation of neutralizing antibody resided on a 52,000-molecular-weight (52K) glycoprotein. The envelope of HSV contains several glycoproteins: one component at 59K and a complex of two or three components at 130K, none of which corresponds in molecular weight to gp52. Using the antisera as immunological probes, we performed pulse-chase experiments with [(35)S]methionine-labeled HSV-1-infected cells and followed the disposition of the glycoproteins during the infectious cycle. Each antiserum immunoprecipitated a (35)S-labeled 52K protein from lysates of cells pulse-labeled at 5 h after infection. By 10 h, the label was chased into a 59K protein also precipitable by each of the three antisera. The results suggest that gp52 is a precursor of gp59 and that the latter corresponds in molecular weight to one of the major glycoproteins of the virion envelope.


Inhibition of reverse transcription of 70S and 35S avian myeloblastosis RNAs by nonprimer tRNA's. We studied the kinetics of the reverse transcription of 70S and 35S RNA of avian myeloblastosis virus in the presence and absence of various tRNA's. All tRNA's inhibited synthesis. tRNA's from Escherichia coli and yeast exhibited a noncompetitive type of inhibition, i.e., they bound reversibly and randomly and did not alter the affinity of the viral RNA for the polymerase. Nonprimer tRNA's obtained from 70S RNA molecules produced a complex pattern of inhibition. The results show that the nonprimer tRNA's which bound to the reverse transcriptase decreased the affinity of the viral RNA for the enzyme. The maximum rate of synthesis with 70S RNA as the template was less than that with 35S RNA, presumably because the former contains nonprimer tRNA's which can interact with the polymerase.


Induction of type C RNA virus from cultured rabbit lymphosarcoma cells. Type C RNA virus(es) was readily released from primary lymphosarcoma cell cultures of WH/J rabbits after induction with halogenated pyrimidines. The virus contained an RNA-directed DNA polymerase and the p30 structural protein. The rabbit virus RNA-directed DNA polymerase and p30 protein shared antigenic homologies with other mammalian type C oncornaviruses but appear to also possess unique antigenic determinants. Normal rabbit liver contained DNA homologous to a 3H-labeled complementary DNA transcript of the rabbit viral genome, indicating that type C viral genetic information is present in at least the WH/J strain of rabbits.


Origin and biological properties of a new BALB/c mouse sarcoma virus. A focus-forming virus previously isolated from a BALB/c mouse hemangiosarcoma has been shown to be replication defective. Analysis of individual BALB/c mouse sarcoma virus (BALB-MSV) nonproducer transformants for expression of helper virus-coded proteins revealed genetically stable variants that expressed two, three, or all four gag gene products in the absence of detectable helper viral env gene expression. The type-specific antigenic determinants of helper viral proteins encoded by the BALB-MSV genome and by the B-tropic virus isolated from the BALB-MSV stock were demonstrated to be indistinguishable from those of BALB:virus-1, a known endogenous virus of BALB/c cells. These findings imply that a BALB/c endogenous virus was involved in the generation of BALB-MSV. By the same immunological approach, the presence of at least a portion of the Moloney-MuLV gag gene has been identified in two other transforming viruses--Moloney-MSV and Abelson lymphosarcoma virus--previously isolated from the BALB/c strain. The tissue culture properties of cells transformed by these defective viruses were also shown to be distinguishable. These findings indicate that transforming virus isolates of the same inbred strain differ in their transforming activities as well as in the helper viral sequences stably associated with their genomes.


Evidence for membrane potential changes in isolated synaptic membrane ghosts monitored with a merocyanine dye. Changes in the fluorescence intensity of merocyanine-540 were measured in suspensions of synaptic plasma membrane ghosts isolated from rat brain cortex. With preincubation of the membrane ghosts in isotonic KCl or NaCl solution, K- and Na-enriched ghosts samples were prepared. In suspensions of both sort of synaptic membrane ghosts, merocyanine-540 showed a fluorescnece emission peak at a wavelength of 590 nm. Under a fixed total concentration of NaCl and KCl, high external K+ induced an increase in the fluorescence intensity, such an increase being proportional to logarithm of K+ concentration. Replacing K+ by Rb+, NH4+ or Cs+, a similar effect was observed. Rb+ was about as effective as K+ ; NH4 is about 2/3 and Cs+ 1/4 as effective. But the changes in fluorescence with increasing K+ concentration were larger in K-ghosts than in Na-ghosts. K+-induced fluorescence changes were very small when gramicidin D was added to the suspension. K+-induced fluorescence changes were not observed in ultrasonicated ghost suspensions. Such findings seem to indicates that the K+-ions induced fluorescence increase reflect the depolarization in the isolated synaptic plasma membrane ghosts. Furthermore, the permeability ratio PNa/PK (PNa, PK: the permeability constant for Na+ and K+) was estimated to be smaller than 0.03.


[Cardiac arrhythmias in exposure to +Gz loads after immersion]. The frequency and pattern of cardiac arrhythmias were studied during an exposure to acceleration +3 Gz for 5 min before and after 3- and 13-day dry immersion. Following immersion the frequency of extrasystolic arrhythmias increased four-fold and more severe arrhythmias--group and systemic extrasystoles--developed as compared with control exposures. The following factors--increased in the parasympathetic tone, disorders in the systemic and regional circulation and fluid-electrolyte balance--seemed to be mainly responsible for distrubed regulation of the cardiovasucular rhythm during a post-immersion exposure to acceleration.


Management of malignant pleural effusion. A pleural effusion is a frequent complication of malignant disease. Essential to the care of oncology patients is a fundamental knowledge of the pathophysiology and treatment of such effusions. This article discusses the current thoughts concerning the occurrence of malignant effusions, outlines the current available methods and agents employed for control, and presents a modification of the thoracostomy procedure that appears to be more effective than the standard procedure.


Scabies and pediculosis.

Scabies and pediculosis. There has been a recent notable increase in scabies and pediculosis pubis and pediculosis capitis. Once the diagnosis is established (preferably by demonstrating the mite or louse), therapy includes consideration of a risk to benefit ratio, explanation of the nature of the infestation to the patient, application of appropriate therapeutic techniques, and follow-up procedures. Resistance of the Sarcoptes mite to modern scabicides has been suggested but not proved. Resistance of the head louse and body louse to modern pediculicides has been reported in other countries but not in the United States.


Antimicrobial resistance and enterotoxin production among isolates of Escherichia coli in the Far East. The frequency of association between transferable extrachromosomal D.N.A. (plasmid) mediated antibiotic resistance and enterotoxin productin is unknown. The antimicrobial susceptibility of 176 enterotoxigenic Escherichia coli from 57 children and adults in the Philippines, Korea, Taiwan, and Indonesia has been examined. 126 isolates (72%) were resistant to one or more antibiotic(s); 77 (44%) were resistant to four or more antibiotics. 43 E. coli which produced both heat-labile and heat-stable toxin, 110 isolates which produced only heat-labile toxin, and 23 which produced only heat-stable toxin were frequently resistant to multiple antibiotics. 25 of 31 resistant isolates tested, 80% transferred antibiotic resistance in bacterial mating experiments. In 35% of the matings transferring antibiotic resistance, the ability to produce enterotoxin was also conferred on the recipients. This in-vitro observation suggests that the widespread use of antibiotics could increase the distribution of enterotoxigenic E. coli, as genes coding for antibiotic resistance and enterotoxin production are frequently transferred together.


Percutaneous liver biopsy and chronic liver disease in haemophiliacs. Systematic screening of forty-seven haemophiliacs in Sheffield revealed abnormal liver-function tests in thirty-six (77%), with a tendency for these abnormalities to persist. To assess the importance of these abnormalities, percutaneous liver biopsy was carried out on eight symptom-free patients under factor-VIII cover. A wide spectrum of chronic liver disease was demonstrated, including chronic aggressive hepatitis and cirrhosis. The liver pathology bore no relation to clinical history or to biochemical findings. Hepatitis-B-virus markers were common, but evidence suggests that this is not the only factor contributing to the development of liver disease. The high incidence of chronic liver disease seems to be a recent development and is probably related to factor-concentrate replacement therapy.


Comparison of tienilic acid with cyclopenthiazide in hyperuricaemic hypertensive patients. Tienilic acid, a diuretic with uricosuric properties, was compared with cyclopenthiazide, in an open, random-order, within-patient crossover study (3 months on each drug) in 36 hyperuricaemic hypertensive patients. All were on an established dose of cyclopenthiazide; most were also on a beta-blocker which they continued to take in their usual dose. A mean dose of 210 mg of tienilic acid gave the same antihypertensive and diuretic effect as a mean dose of 0.41 mg of cyclopenthiazide. Serum uric acid was very much lower when patients were on tienilic acid (0.29 mmol/l) than on cyclopenthiazide (0.50 mmol/l). Apart from slightly higher serum-chloride and serum-urea during the period on tienilic acid, no major differences in serum-electrolytes, renal-function tests, glucose tolerance, and fasting lipids were observed. Audiometric tests showed that tienilic acid was not ototoxic. S.G.O.T. and S.G.P.T. rose to pathological values in 3 women when they were on tienilic acid, to a lesser extent, in 2 men when they were on cyclopenthiazide. There is no definite evidence that the changes in the transaminases were related to tienilic acid. Some postural hypotension or slight fluid retention occurred during the initial, dose-finding period, and 3 patients had mild indigestion but no patient had to discontinue the trial because of side-effects.


Clonidine blocks acute opiate-withdrawal symptoms. In a double-blind, placebo-controlled, cross-over trial, clonidine eliminated objective signs and subjective symptoms of opiate withdrawal for 240--360 min in eleven addicts in a hospital setting. In an open pilot study of the effects of clonidine on longer-term opiate abstinence and symptoms, the same patients did well while taking clonidine for one week. There was only one documented instance of heroin use, in a patient who did not take clonidine after hospital discharge. 6 weeks or more after the study, four patients were back on reduced doses of methadone, one was on tricyclic antidepressants, and seven were off of all opiates. All eleven patients were doing well. These data suggest that opiate withdrawal is due to increased neuronal activity in areas such as the locus coeruleus which are regulated by both alpha-2 adrenergic and opiate receptors.


A syndrome of hypertension, convulsion, and cerebral haemorrhage in thalassaemic patients after multiple blood-transfusions. Various combinations of hypertension, convulsion, severe headache, and cerebral haemorrhage appeared in eight thalassaemic patients after they had received 3--7 units of blood in preparation for splenectomy. Intracranial haemorrhage definitely developed in four patients, three of whom died. At necropsy the brains showed changes similar to those of hypertensive cerebral haemorrhage and hypertensive encephalopathy. It is believed that hypertension initiates this syndrome. Since the episodes often occurred days, as long as 15 days, after the last unit of blood was transfused, hypertension did not seem to result from volume overload, but probably from vasopressive substances provided by or occurring in association with multiple blood-transfusions. Host factors may also contribute.


Cephalothin plus an aminoglycoside is more nephrotoxic than methicillin plus an aminoglycoside. In a prospective, randomised, double-blind trial to determine if cephalothin plus an aminoglycoside is more nephrotoxic than methicillin plus an aminoglycoside, patients were assigned to one of four treatment groups: cephalothin and gentamicin (C.G.), cephalothin and tobramycin (C.T.), methicillin and gentamicin (M.G.), or methicillin and tobramycin (M.T.). The incidence of definite nephrotoxicity was: C.G., 7/23 (30.4%); C.T., 5/24 (20.8%); M.G., 2/20 (10%); and M.T., 1/23 (4.3%). There was no statistically significant difference in nephrotoxicity between the combined gentamicin groups (C.G. and M.G.) and the combined tobramycin groups (C.T. and M.T.). Definite nephrotoxicity developed in 12/47 (25.5%) of the combined cephalothin groups (C.G. and C.T.) and in only 3/43 (7%) of the combined methicilllin groups (M.G. and M.T.). The combination of cephalothin plus an aminoglycoside is therefore more nephrotoxic than the combination of methicillin plus an aminoglycoside.


Abnormal rectal immunoglobulin pattern in Hirschsprung's disease. The immunoglobulin content of rectal biopsy tissue and secretions from twelve neonates in whom Hirschsprung's disease was suspected was determined by an organ-culture technique and radioimmunoassay. The immunoreactive IgG content of explanted rectal tissue and its secretions in those six children who proved to have Hirschsprung's disease was much higher than in those with other types of obstructive lower-intestinal disease. Increased amounts of IgG may represent maternally derived antibody associated with neonatal gut neural elements. This seems to be the first report of an immunological abnormality in Hirschsprung's disease.


Role of mononuclear infiltrating cells in pathogenesis of hepatitis. Mice that have been injected with Corynebacterium parvum have mononuclear-cell infiltrates in the liver lobules. In such mice a small dose of lipopolysaccharide endotoxin produced a lethal hepatitis, with high serum-transaminase concentrations, glycogen depletion, and hypoglycaemia. It is suggested that lipopolysaccharide triggers the release from the infiltrating mononuclear cells of factors toxic for hepatocytes. Similarly certain parasitic and virus infections and graft-versus-host reactions can sensitise mice to the induction of hepatitis by exposure to small doses of lipopolysaccharide. This model may be applicable to human hepatitis.


Colitis.

Colitis. The disability caused by proctitis or colitis has been assessed among patients attending a hospital outpatient clinic. Although bowel frequency was a common and troublesome symptom, urgency of defaecation with a tendency to precipitate incontinence was a major factor limiting working life and leisure and social activities. Abdominal or rectal pain and lassitude were the other main symptoms. In every aspect of life studied the disability was as great for patients with proctitis or distal colitis as for those with more extensive inflammation of the colon. As a result of symptoms about a fifth of the patients had a reduced earning capacity and quarter were restricted in their social or leisure activities even when symptoms were at their least bad. The social disability of proctitis or colitis may be underestimated because patients do not mention their fear of incontinence and because their complaint of lassitude does not always correlate with the apparent activity or extent of the disease.


The continuing risk of vitamin-D intoxication. Eight cases of vitamin-D poisoning are described. In six patients this therapy was unnecessary and in the remaining two patients inadequate supervision of the treatment resulted in overdosage. In five cases the history of vitamin-D therapy was either unknown or not appreciated by the referring clinician so that three surgical procedures of dubious value were performed on two patients. Large doses of vitamin D should only be used when strictly indicated and on the understanding that close biochemical and clinical supervision is necessary.


Breast-milk monitoring to measure Michigan's contamination with polybrominated biphenyls. In 1973 and 1974, several thousand Michigan dairy farms were contaminated by polybrominated biphenyls (P.B.B.) as the result of an industrial accident. An unknown quantity of contaminated meat and dairy products entered the food chain before contaminated farms were quarantined. To determine the extent of human exposure, P.B.B. concentrations were measured in human breast milk, which was collected in a random-sample survey from nursing mothers throughout Michigan. 96% of 53 samples from Michigan's lower peninsula and 43% of 42 samples from the less densely populated upper peninsula contained detectable levels of P.B.B. These data indicate that about 8 million of Michigan's 9.1 million residents have detectable body burdens of P.B.B.


Aminoglutethimide in treatment of metastatic breast carcinoma. 42 patients with metastatic breast carcinoma were treated with aminoglutethimide, which inhibits adrenal steroid hormone synthesis. Treatment was stopped in 2 patients before response could be assessed; of the other 40, 15 (37.5%) had an objective response, 1 (2.5%) showed a response in bone but not in soft tissue, and 4 (10%) had complete or very great relief of metastatic bone pain but no radiological evidence of improvement. 19 (53%) of 36 patients with bone metastases responded to treatment (15 had X-ray evidence and 4 had pain relief), as did 5 (45%) of 11 patients with soft tissue metastases, 2 (25%) of 8 with malignant marrow infiltration, 1 (14%) of 7 with lung metastases, and none of 13 with liver metastases. Response was commonest in patients who had previously responded to other forms of endocrine therapy. Side-effects, usually mild and transient, occurred in a few patients; the most important were an initial period of somnolence in 9 patients and a rash in 5.


Response of plasma-25-hydroxyvitamin D to ultraviolet irradiation in long-stay geriatric patients. The response of plasma-25-hydroxyvitamin D (25[OH]D) to different exposures to ultraviolet irradiation has been studied in patients in long-stay geriatric wards. Increases sufficient to bring the plasma-25(OH)D into the normal range may be obtained with doses less than those required to produce erythema. The provision of such background irradiation may be a suitable method of preventing vitamin-D deficiency in elderly subjects who receive very little exposure to sunlight.


Hepatitis B outbreak among chimpanzees at the London Zoo. Persistent carriage of hepatitis B virus in extremely high titre was identified in 5 out of 9 chimpanzees kept at the London Zoo. Antibody to this virus was present in the other 4 chimpanzees. Serological survey of the other primates in the Regent's Park collection did not reveal the presence of the surface antigen in 2 gorillas, 11 orang-utans, and 2 gibbons, although surface antibody was present in the serum of 1 gorilla and 2 orang-utans. 3 of the carrier chimpanzees were born at the Zoo and were the offspring of either a carrier mother or a carrier father, and perinatal transmission may have occurred. A strict safety code of practice was introduced and hepatitis B immunoglobulin was given at intervals to designated staff members. Sero-conversion did not occur in any of the 38 staff members under surveillance for more than 2 years. Treatment of the carrier state in the chimpanzees was attempted with human leucocyte interferon, with and without ribavirin ('Virazole'), and with adenine arabinoside, but the effects were mostly temporary.


The mechanism of placebo analgesia.

The mechanism of placebo analgesia. The effect of naloxone on dental postoperative pain was studied to examine the hypothesis that endorphins mediate placebo analgesia. All patients had extraction of impacted mandibular third molars with diazepam, N2O, and local block with mepivacaine. 3 h and 4 h after surgery naloxone or a placebo was given under randomised, double-blind conditions. Pain was evaluated on a visual analogue scale. Patients given naloxone reported significantly greater pain than those given placebo. Patients given placebo as their first drug was either placebo responders, whose pain was reduced or unchanged, or nonresponders whose pain increased. Naloxone given as a second drug produced no additional increase in pain levels in nonresponders but did increase pain levels of placebo responders. Nonresponders had a final mean pain rating identical to that of responders who received naloxone as their second drug. Thus the enhancement of reported pain produced by naloxone can be entirely accounted for by its effect on placebo responders. These data are consistent with the hypothesis that endorphin release mediates placebo analgesia for dental postoperative pain.


Repeated failure of nickel-containing prosthetic heart valves in a patient allergic to nickel. Life-threatening peri-prosthetic incompetence developed with two successive nickel-containing mitral-valve prostheses in a patient allergic to nickel. Neither prosthesis had been incorporated satisfactorily. Her present nickel-free prosthesis seems to be satisfactory 22 months after insertion. Since allergy to nickel may have been involved in the failure of these prostheses, it is recommended that nickel-sensitive patients should be given nickel-free prostheses.


Mitochondrial inheritance and disease.

Mitochondrial inheritance and disease. Spontaneously occurring variants of the D.N.A. content of mitochondria may be responsible for human disease. Among the prime candidates for such a mitochondrial aetiology are certain drug-induced blood dyscrasias, particularly that due to chloramphenicol. Because mitochondria are generally inherited from the female parent, such disorders should be clustered among matroclinally related individuals. The clinical manifestations of such diseases are a function of the manner in which mitochondria are allocated to somatic cells and tissues during development.


Mycobacteria as a possible cause of inflammatory bowel disease. Mesenteric lymph-nodes from 27 patients with Crohn's disease, 13 with ulcerative colitis, and 11 without inflammatory bowel disease were cultured for mycobacteria. A node from a patient with Crohn's disease yielded a strain of Mycobacterium kansasii. Cultures from 22 other patients with Crohn's disease, 7 with ulcerative colitis, and 1 control subject yielded pleomorphic organisms with the electron-microscopic appearances of cell-wall-deficient organisms. Further culture and characterisation of these organisms has so far proved unsuccesful. Skin tests with tuberculin were positive in a smaller proportion of patients with Crohn's disease than in healthy control subjects. Conversely, the patients gave a higher proportion of positive reactions to a reagent prepared from the strain of M. kansasii isolated. No differences in the proportion of positive test were found between patients and controls with reagents prepared from 16 other mycobacteria. Cell-wall-deficient mycobacteria are a possible causative agent of inflammatory bowel disease.


Search by immunofluorescence for antigens of Rotavirus, Pseudomonas maltophilia, and Mycobacterium kansasii in Crohn's disease. Crohn's-disease tissue was tested by indirect immunofluorescence for antigens of rotavirus, Pseudomonas maltophilia, and Mycobacterium kansasii. No reactions were obtained with anti-serum to P. maltophilia and M. kansasii, and a granular fluorescence seen with rotavirus antibody was probably non-specific.


Subcutaneous ancrod in prevention of deep-vein thrombosis after operation for fractured neck of femur. In a randomised double-blind controlled trial 53 patients received 5 daily subcutaneous injections of ancrod ('Arvin') after operation for fractured neck of femur, and 52 patients received saline fractured neck of femur, and 52 patients received saline injections. Deep-vein thrombosis (D.V.T.) was detected by bilateral ascending venography or necropsy 6--16 days after surgery. The frequency of D.V.T. and bilateral D.V.T. was significantly lower in the ancrod group (P less than 0.01). The frequency of major D.V.T. (thrombi in veins proximal to the calf, or calf-vein thrombi more than 3 cm long) was also significantly lower in the ancrod group (P less than 0.001). No complications of ancrod prophylaxis occurred. Ancrod reduced plasma-fibrinogen, and hence plasma and blood viscosity, during the first week after surgery; preoperative levels of fibrinogen and viscosity were not associated with post-operative D.V.T. Subcutaneous ancrod is a simple and effective alternative to oral anticoagulants for the reduction of the frequency of D.V.T. after operation for hip fracture, and merits assessment in other high-risk groups of patients.


Deterioration of renal function during treatment of chronic renal failure with 1,25-dihydroxycholecalciferol. A controlled study of the effects of the potent vitamin-D metabolite, 1, 25-dihydroxycholecalciferol (1,25[OH]2D3), and vitamin D3 was done in 18 non-dialysed patients with chronic renal failure (C.R.F.). Patients with a creatinine clearance below 35 ml/min and mild renal osteodystrophy were selected. After 6 months' observation of the spontaneous course the patients were randomly allocated to 6 months' oral treatment with either 1, 25 (OH)2D3 or vitamin D3 in initial daily doses of 1microgram and 4000 I.U., respectively, combined with 0.5 g calcium. 1,25(OH)2D3 quickly corrected hypocalcaemia, reduced serum-alkaline-phosphatases and serum-immunoreactive-parathyroid-hormone, and more than doubled the urinary excretion rate of calcium. D3 had similar, but less pronounced effects. 7 out of 8 patients on 1,25(OH)2D3, developed hypercalcaemia which necessitated a reduction in dosage. None of the patients on D3 treatment developed hypercalcaemia. The percentage fall in creatinine clearance was greater during treatment than before treatment in all patients on 1, 25 (OH)2D3 (P less than 0.01) and in 7 of 9 patients on vitamin D3 treatment (though the group change here was not significant). Deterioration of renal function is a major limitation of the clinical use of 1, 25(OH)2D3 and D3 in non-dialysed patients with C.R.F. In fact, the decrased formation of 1, 25(OH)2D3 seen in C.R.F. might protect renal function at the expense of abnormalities in mineral metabolism.


Innappropriate renin secretion unmasked by captopril (SQ 14 225) in hypertension of chronic renal failure. Captopril (SQ 14 225), an orally active inhibitor of angiotensin-converting enzyme, was given to 7 hypertensive patients with chronic renal failure whose plasma-creatinine ranged from 1.5--7.4 mg/dl; whose plasma-renin activity was normal; whose hypertension was not controlled by previous therapy consisting in 5 patients of three or more antihypertensive drugs; and whose blood-pressures averaged 176/111 +/- 11/3 mm Hg. Inhibition of converting enzyme by oral captopril, 200 mg twice daily, reduced blood-pressure to 156/100 +/- 9/5 mm Hg. 5 patients needed additional treatment by frusemide 40--250 mg/day orally. With this combined regimen the blood-pressure of all patients averaged 126/85 +/- 4/3 mm Hg after 8 +/- 2 weeks of captopril. The drug was well tolerated. These results suggest that inhibition of angiotensin-converting enzyme with or without sodium depletion is an efficient treatment for hypertension associated with chronic renal failure. It appears that although renin levels in patients with this condition may be "normal", they are inappropriate in relation to the subtle degree of sodium retention that occurs with this disorder.


Acute colitis related to penicillin and penicillin derivatives. 5 cases of acute transient colitis associated with the ingestion of ampicillin, an ampicillin derivative, and penicillin are discussed. The clinical presentation, course, and radiographic and sigmoidoscopic findings are distinct from those in pseudomembranous colitis. The colonic changes may be related to an allergic reaction in the intestine which results primarily in transient ischaemia. The presenting symptom is blood diarrhoea, and barium enema is the most productive diagnostic examination.


Evidence of acquired immune deficiencies in Mediterranean lymphoma. A possible aetiological link. Patients with Mediterranean lymphoma (M.L.) had a significant reduction in humoral immunity (IgG and IgM) as well as impaired cellular immunity (50% were anergic to three antigens--P.P.D., mumps, and dinitrochlorobenzene). Any hypothesis for the pathogenesis of M.L. has to account for the peculiar geographic distribution of the disease, the age and sex incidence, the plasma-cell nature of the tumours, the associated heavy plasmacytic proliferation with relatively intact intestinal mucosa, involvement of the proximal small intestine, and alpha-chain production in a large proportion of patients. All areas in which M.L. is common are currently involved in the seventh cholera pandemic. Vibrio cholerae toxin inhibits both immediate and delayed immune reaction in vitro through its effect on cyclic adenosine monophosphate. V. cholerae antigens stimulate the proliferation of IgA-producing immunocytes in the mucosa without deeply penetrating the mucosa. The proximal small bowel is usually affected by the disease but there is little epithelial damage. The population in endemic areas is continuously exposed to V. cholerae antigens and toxins. It is suggested that such exposure, under certain genetic or other circumstances, may produce a state of immunosuppression in the gut thus accelerating, predisposing to, or producing lymphoplasmacytic neoplasia.


Drug treatment of epilepsy.

Drug treatment of epilepsy. The difficulties of the drug treatment of epilepsy include the high prevalence of the disorder, poor prognosis, prolonged multidrug treatment, chronic toxicity, and uncertainty of the relative efficacy and toxicity of individual anticonvulsants. These problems are reviewed in relation to recent knowledge of the clinical pharmacology of the drugs, the application of which offers the possibility of a more simple, rational, and effective approach to therapy. There is considerable potential for monotherapy assisted by drug-level monitoring.


Fatal echovirus 11 infections in outbreak in special-care baby unit. In December, 1977, an outbreak of echovirus 11 infection occurred in the special-care baby unit of the Cambridge Maternity Hospital. 3 neonates died, and of 24 infants on the ward during the epidemic 6 others were infected, 3 with symptoms. 3 infants had similar symptoms but virus could not be isolated. The pathological changes in the 3 fatal infections, have not previously been reported for echovirus 11; nor have fatal infections occurred in an epidemic in a special-care baby unit.


Natural history of perforated duodenal ulcers treated by suture closure. Of 50 patients with a perforated duodenal ulcer treated with simple suture closure, 36 (72%) had symptoms before the perforation for a mean period of 9.9 years. 24 of the 36 (66.6%) acquired further symptoms or complications at 1--48 months (mean 11 months) after the operation, whereas only 2 of the 14 patients (14%) who were symptom-free preoperatively had further complications. At follow-up 48% of the patients were still free of symptoms. Simple suture closure with a "wait and see" policy remains the treatment of choice, and a prospective clinical study to evaluate definitive emergency surgery for duodenal ulcer has been cancelled.


Controlled trial prednisolone in acute polyneuropathy. In a multicentre, randomised trial of prednisolone in acute polyneuropathy of undetermined aetiology (Guillain-Barré syndrome), 21 patients were treated with prednisolone (60 mg daily for one week, 40 mg daily for four days, and then 30 mg daily for three days) and 19 did not have steroid treatment. Patients were graded on a six-point scale by one of two neurologists who had no knowledge of the treatment schedule. Reassessment at one, three, and twelve months consistently showed greater improvement in the control than the prednisolone group but the only statistically significant result was in the improvement at three months among patients entered to the trial within a week of onset of illness. The 6 control patients had improved by 2.5 +/- 0.43 grades by three months from entry to the trial whereas the 10 prednisolone patients had only improved by 0.9 +/- 0.46 grades (P less than 0.05). There was 1 death related to the polyneuropathy in each group, and 1 suicide in a control patient during convalescence. 6 prednisolone patients were left with considerable disability compared with 1 control patient. There were 3 relapses in the prednisolone group, but none in the control group. The results indicate that steroid treatment is not beneficial and can be detrimental in acute neuropathy of undetermined aetiology.


Systemic lupus erythematosus and lymphoma. In 4 women lymphomas developed 2 months to 12 years after the onset of systemic lupus erythematosus. An association between the two diseases had previously been reported in 14 cases, in 6 of which the lymphoma either preceded or was diagnosed at the same time as the autoimmune disease. In systemic lupus erythematous early biopsy of suspect lymph-nodes is recommended.


Enzymatic basis of typical X-linked icthyosis. Steroid sulphatase activity was determined in cultured fibroblasts from 25 individuals with X-linked ichthyosis from four countries. All those with X-linked disease had markedly reduced enzyme levels compared with controls and patients with other types of ichthyosis. X-linked ichthyosis seems to be the result of a common mutation affecting the expression of steroid-sulphatase activity.


Single lead for permanent physiological cardiac pacing. A method of achieving permanent pacing by the use of a single bipolar transvenous lead is described, the atrial and ventricular electrodes being located on the same lead 14--17 cm apart. The new leads have been implanted successfully in two patients.


Possible diagnostic test for Crohn's disease by use of buccal mucosa. Immunofluorescent tracing was used on buccal mucosa from patients with Crohn's disease to investigate some of its immunological characteristics, and to compare these with those of mucosa from controls. Normal buccal mucosa from patients with Crohn's disease, incubated with its own serum then stained for deposited antibody by the fluorescent technique, showed a positive reaction, not observed in buccal mucosa from normal persons or patients with ulcerative colitis. These observations could provide the basis of a diagnostic test.


Diagnosis of protein-losing enteropathy by gastrointestinal clearance of alpha1-antitrypsin. Alpha1-antitrypsin was assessed, in 10 patients with protein-losing enteropathy and 13 control subjects, as an endogenous marker of plasma-protein loss into the gastrointestinal tract. Both faecal alpha1-A.T. concentrations and faecal alpha1-A.T. clearance were significantly higher in patients than in controls. Wtih clearance there was no overlap between the groups. Over 10 days the normal gastrointestinal clearance of alpha1-A.T. was 3.07 +/- 2.25 (S.D.) ml/day. Measurement of alpha1-A.T. clearance is easy and requires no radioisotopes.


The human rumen.

The human rumen. The cow is a ruminant, and cow's milk has evolved to promote bacterial growth in the upper small bowel; whereas human milk has evolved to discourage bacterial growth. Examination of the constituents of the two milks shows that their differences can be accounted for in terms of this difference in function. Children who are fed a calf's diet tend to develop a rumen. This may lead to chronic diarrhoea and malnutrition and may be a factor in diarrhoea ascribed to cow's-milk-protein allergy and lactose intolerance.


Mortality from carcinoma of cervix uteri in Britain. A review of age-specific mortality-rates from cervical cancer in England and Wales and in Scotland in 1968--76 shows a decline, striking at some ages. In England and Wales, however, there has been an increase at ages 25--34 and, possibly, at 15--24. Signs of such an increase are also seen in Scotland for the age-groups 25--34 when figures for two regions with well-established screening programmes are removed. Indeed trends in these regions compared with those for the rest of Scotland support the benefit of cervical screening.


Cervical cancer in younger women.

Cervical cancer in younger women. Cervical cancer does not begin in women at some vague mid-procreative term, but is a concomitant of early sexual activity. In Brighton the mean age of patients with histologically proven carcinoma of the cervix has been falling, from 50 in 1967 to 35 by 1977. In one year abnormal cells were found in cervical smears from 24 teenage girls (5 aged 16, 2 aged 17, 8 aged 18 and 9 aged 19), and in 4 of these teenage cases malignancy was subsequently proved histologically. Until the aetiology of the  disease is established no lower age limit should be set for cervical screening; all young women should be entitled and encouraged to have cervical cytology tests.


Suppression of serum thyrotropin (TSH) concentrations following thyroidectomy and cold exposure by passive immunization with antiserum to thyrotropin-releasing hormone (TRH) in rats. Administration of antiserum to synthetic thyrotropin-releasing hormone (TRH) to thyroidectomized rats caused a significant depression of serum thyrotropin (TSH). Serum TSH and triiodothyronine (T3) responses to cold exposure (4 +/- 1 C) were abolished by administration of anti-TRH serum. In addition, synthetic TRH lost its biological activity when bound to the gamma globulin fraction from the anti-TRH serum. These observations provide evidence that TRH is involved in the mechanism of enhanced TSH secretion from the pituitary following thyroidectomy and cold exposure.


[Experiences with the electrophoretic mobility test in children. A diagnostic help in pediatric oncology? (author's transl)]. Lymphocyte sensitisation to encephalitogenic factor (EF) was determined in 131 children with the electrophoretic mobility test (EM-test) to find out, whether this test may be helpful in the diagnosis of malignant disease in children. None of 34 healthy controls showed a decrease of electrophoretic mobility of more than 5%, while all 10 children with malignant solid tumors showed a slowing of more than 5%. 3 of 54 patients with non malignant disease showed a slowing of more than 5% in the EM-test. Children with malignant solid tumors during therapy and children with leukemia during different stages of the disease often showed a slowing of less than 5% in the EM-test. The possible diagnostic help of the EM-test is shown in a case history. Finally some technical remarks are made on improving this test, and further studies are suggested.


Responses of radiation-sensitive mutants of Saccharomyces cerevisiae to lethal effects of bleomycin. Haploid and diploid strains of yeast containing genes conferring radiation-sensitivity were studied under growing and nongrowing experimental conditions for their relative sensitivities to growth-inhibitory effects of bleomycin (BM). The rad1, rad2, rad3, rad4, rad5 (and allelic rev2), rad7, rad10, rad11, rad 12, rad14, rad15, rad16 and rev3 strains exhibited responses similar to normal (Rad+) yeast strains. It is concluded from these findings that the excision-repair function deficient in several of these mutant strains is not important for repair of bleomycin-induced damages in yeast. The sensitive strains contained rad6, rad9, rad18, rad22, rad50, rad51, rad52, rad53, rad54, rad55, rad56, rad57 and rs1. Strains bearing rad8 or rad19 could not be classified unambiguously. With one exception, all rad mutants found very sensitive to BM were sensitive to X-rays, suggesting that some aspect of the repair of BM- and X-ray-induced damages in yeast may be similar. Sensitivities to BM and radiation co-segregated in pedigrees following meiosis, and several BM-resistant revertants isolated from two rad6 mutant strains sensitive to BM, X-rays and UV were cross-resistant to all three agents. These results confirm that the rad mutants were responsible for the cross-sensitivities in the original strains.


[Protein fractions and total proteins in serum of rats on high fat diets]. This paper deals with the behaviour of serum protein fractions in rats fed 22 resp. 52 weeks a 50% or 3% fat diet using Millipore electrophoresis. The total serum protein concentration was also estimated. In rats on the high-fat diet the albumin fraction significantly dropped, while elevated levels of alpha 1- and gamma-globulin fraction were found. Serum protein content was slightly elevated in rats fed the diet 22 weeks, but this elevation was only significant after 52 weeks feeding. Within the same diet group there were no significant differences observed.


Synthesis of an ovalbumin-like protein by Escherichia coli K12 harbouring a recombinant plasmid. A cloned DNA transcript of ovalbumin mRNA was cut a few nucleotides away from the initiator codon, and fused in phase to the beginning of the Escherichia coli beta-galactosidase gene. The hybrid gene has been cloned in E. coli where it produces large amounts of an ovalbumin-like protein.


Effects of 5-aza-2'-deoxycytidine on DNA synthesis in mouse lymphatic tissues. 5-Aza-2'-deoxycytidine-3H administered i. p. to mice is preferentially incorporated into nucleic acids in the spleen and thymus. The labeling of the spleen is maximal during first hours after drug administration and decreases rapidly thereafter. Following administration, 5-aza-2'-deoxycytidine is metabolized giving rise to several new compounds. The incorporation of 5-aza-2'-deoxycytidine is blocked by the simultaneously administered deoxycytidine to a lesser degree than that of deoxycytidine by the analogue. Maximal inhibition of deoxycytidine incorporation by the drug was observed in the spleen; the incorporation of thymidine under similar conditions was diminished only about 20%. However, pretreatment of the animals with the analogue resulted in a strong inhibition of thymidine incorporation (about 80%) both in the spleen and the thymus. Simultaneously the activity of thymidine and thymidylate kinases in cell-free spleen extracts was markedly depressed.


The retrovirus particles in human myeloma cells RPMI8226: morphological, biochemical, immunological and infective transmission studies. The retrovirus designated RPMI8226V (isolated from human myeloma cells RPMI8226) has been characterized with respect to its morphological, biochemical and immunological properties as well as its propagation in various animal and human cells. The myeloma cells RPMI8226 produce intracytoplasmatic A-type particles and extracellular particles. The extracellular particles have been classified as immature particles with translucent core center, typical mammalian C-type virus particles and C-type particles with intermediate membrane. However, the budded particles in secondarily infected human neoplastic cells contained complete doughnut-shaped nucleoids. This type of budding is rather characteristic for B-type particles. The 3H-uridine labeled RPMI8226 viral particles have a buoyant density 1.17 g/ml in sucrose gradient containing high molecular weight RNA and the distribution of viral structural proteins in SDS-PAGE is characteristic for oncornaviruses. The internal structural proteins according to MW are ranged from 13 000 to 30 000 daltons. The virus contains a magnesium-dependent reverse transcriptase. The cellular homogenate and viral concentrate from RPMI8226 cultures do not react with antibodies against ALSV, MuLV, FeLV, RD114, MP-MV and SiSLV. The only reaction was scored with anti BLV antibodies. However, anti BLV serum inhibiting the reverse transcriptase activity of BLV to 60% does not cross-react with the reverse transcriptase of RPMI8226V. In contrast to BLV concentrates, neither XC nor KC cells show syncytia formation by RPMI8226V. The RPMI8226V replication is restricted to human tumor and normal human glia-like cells. The possible origin of the virus is discussed.


Checking of carcinoma patients with the leukocyte migration technique (LMT) under agarose. Leukocyte migration tests under agarose (Clausen technique) were performed in 28 patients tentatively diagnosed as having any malignancy with the use of a 3 M KCl-extract panel prepared from bronchogenic, gastric, colonic, renal, and mammary carcinoma, corresponding normal tissues, carcinoembryonic antigen (CEA), and human encephalitogenic protein (HEP). 17 out of 22 proven carcinoma patients showed sensitization by reaction with optimal concentrated KCl-extract of cancer from the same organ type as their own tumor. In some cases positive reactions could be observed also with normal tissue antigen (NTA) of tumor organ type (7/22) or with an additional carcinoma extract of organ type differing from patients own primary tumor (8/22). Gastrointestinal carcinomas, especially, showed sensitization to CEA (7/12) contrary to nongastrointestinal carcinomas (1/10). With HEP no positive reactivity could be found (0/10). With the use of tumor antigen panel (5 antigens) only few positive reactions (MI less than 0.80 or greater than 1.20) could be observed in 6 patients with nonmalignant diseases (1/30 tests) and 8 healthy blood donors (1/40 tests). A widespread individual screening program using tissue antigens for patients suspected of malignancies could give a pattern of reactivities and improve the recognition of cell-mediated sensitization against tumor tissues.


[Treatment of malignant lesions of the esophagus with the thermoprobe]. In inoperable stenosing tumours of the oesophagus the problem arises of maintaining oral food intake. As malignant cells are more cold-sensitive than normal cells oesophageal tumours may  be "cold-coagulated" with special probes. Preliminary results with this treatment in 28 patients are reported.


Prenatal genetic diagnosis: nine years experience. The advent of techniques for the prenatal diagnosis of disease represents one of the most important advances in clinical genetics of the past decade. It has made it increasingly possible for parents to make informed decisions about reproduction. Physicians should think of genetic amniocentesis as one more method of assessing the status of a fetus at risk. It should be undertaken for a medical indication only, but the safety and accuracy are sufficiently established for it to be considered an integral part of providing high risk obstetrical care.


Neural tube defects: maternal serum screening and prenatal diagnosis. Neural tube defects represent some of the most common and serious of the congenital malformations. Although elevation of alpha-fetoprotein in amniotic fluid is not diagnostic, it does indicate an abnormality of the fetus in a very high proportion of cases. A normal level, however, does not exclude the possibility of a closed neural tube defect. It is therefore recommended that all amniocenteses performed between 15 and 20 weeks of gestation include measurement of alpha-fetoprotein. Maternal serum alpha-fetoprotein assay is a screening test and pilot studies will be necessary to determine its value as a routine prenatal blood test.


Some guidelines for uses of hypnotherapy in pediatrics. Hypnotherapy has many uses in pediatrics, and its value, not only as a adjunct but also as a primary therapy for certain conditions, justifies its inclusion in pediatric training programs. Suggestion and expectation have long been related to therapeutic outcomes in medicine, but not all physicians know how to apply them constructively and systematically in communication with patients. In pediatrics there is a tendency to overlook opportunities in which hypnosis might be the treatment of choice. Because children engage in imagination and fantasy easily without the cognitive inhibitions of adults, they are able to use hypnosis more readily than adults. More research into the imaginative skills of children may facilitate understanding of learning mechanisms and make it possible for professionals to prevent the loss of the natural imaginative capacities in children and, therefore, enhance the ability of mature members of society to use these skills. In addition to reduction of specific symptoms through hypnotherapy, children benefit by the sense of mastery which they acquire, a sense which is surely needed to overcome the feelings of hopelessness, loss of control, and depression induced by many diagnostic and therapeutic procedures in medicine.


Sequence specific cleavage of DNA by the antitumor antibiotics neocarzinostatin and bleomycin. We have investigated the sites of DNA damage by the antitumor antibiotics neocarzinostatin and bleomycin by using a 5'-end-labeled DNA fragment of defined sequence as a substrate. At the high drug concentrations used here, neocarzinostatin creates single-strand breaks in DNA at positions of adenine and thymine in the presence of 2-mercaptoethanol, and bleomycin cleaves DNA at GC and GT sequences and to a lesser extent at TA sequences with its degradative activity enhanced by 2-mercaptoethanol. In the presence of ferrous ions, bleomycin cleaves DNA at TT, AT, and TA, as well as at GC and GT sequences. Both antibiotics make double-strand breaks in DNA at specific sites and it is likely that these result from two independent single-strand breaks at nearby sites on opposite strands of the DNA.


Association of mitochondria with microtubules in cultured cells. By indirect immunofluorescence techniques, microtubules and mitochondria were localized in normal rat kidney cells, human WI38 fibroblasts, mouse peritoneal macrophages, and a putative smooth muscle rat cell line, in monolayer culture. The mitochondria were found to be arranged along the cytoplasmic microtubules in each cell type. Disruption of the microtubules with colcemid caused a redistribution of the mitochondria in these cells. There was no correlation between the location of the mitochondria and actin-containing filaments. This evidence suggests that mitochondria are directly or indirectly associated with microtubules in these cells.


Immunoglobulin levels in large calf agglomerations. The zinc sulphate turbidity test was used to establish the total immunoglobulin levels of calves from birth to 4 months of age. Colostral immunity fell rapidly and the lowest levels were found in the third and fourth week of life. During the fifth week there was a significant increase. 25% of animals were hypogammaglobulinaemic in the first week of life. This ratio increased to 50% in the third week of life. At the age of two months the lowest immunoglobulin levels almost vanished. These findings are discussed as basic information for the appropriate management of calf agglomerations. Animals should be agglomerated only after the first months of life. When this is not possible, the pens for large agglomerations should have individual housing for the newly admitted calves. The importance of appropriate epizootological measures to reduce infections are pointed out.


Massive postoperative facial edema in a rhytidectomy. Case report. We present a case of massive facial edema secondary to an uncomplicated face-lift procedure, and we discuss the mechanisms of histamine release.


Parental symbolism in the doctor image of asthmatic patients. In a group of asthmatic adult subjects (n = 88) the maternal and paternal aspects in the doctor image have been invesigated. The Semantic Differential Parental, an item list consisting of 18 paternal and 18 maternal items, was employed. With this item list the following concepts were scored: the doctor, the father, the ideal father, the mother, and the ideal mother. In the formation of the doctor image, the contribution of maternal and paternal qualities were both qualitatively as well as quantitatively studied and compared with the parental images. The acquired data are interpreted in comparison with the findings of former studies, with the same methodology, in groups of neurotic, schizophrenic and mixed psychosomatic patients. It appears that asthmatic patients consider the doctor mostly as a minimally differentiated omnipotent figure, possessing all parental qualities, but excelling by paternity, instrumental and nurturance functions. We notice a close likeness with the doctor image of the mixed group of psychosomatic patients. In contrast to neurotics, who project on the doctor more the norm giving pole of paternity and the receptivity pole of maternity, for asthmatics and psychosomatic patients in general the doctor figure is invested with the all-embracing power qualities of both parents.


Quantification of gastrin-producing cells (G cells) and parietal cells in relation to histopathological alterations in resected stomachs from patients with peptic ulcer disease. The distribution and numbers of G cells and of parietal cells were related to the distribution and severity of histopathological alterations (inflammatory cell infiltration, atrophy and intestinal metaplasia) in corresponding mucosal tissue blocks from resected stomachs (12 patients with gastric ulcer, 11 with duodenal ulcer, and 14 with duodenal ulcer and uremia). In all patients the histopathological features were more severe in the pyloric antrum than in the body, and the change in severity corresponded well with the disapperance of G cells at the body-antrum border. The transitional body-antrum zone was histopathologically similar to the remaining antrum. A marked individual heterogeneity of the histopathological alterations was observed. An increasing grade of atrophy was associated with increased severity of inflammation, and the presence of intestinal metaplasia was especially associated with atrophy. No significant correlation was found between the antral G-cell number and the grade of antral inflammatory cell infiltration, whereas there was a reduction in cell number with increasing grade of atrophy in all patient categories. The parietal-cell density in the body mucosa was decreased with increasing grade of inflammation as well as with increasing grade of atrophy. The presence of patchy intestinal metaplasia resulted in a complete absence of G cells and of parietal cells from the corresponding part of the mucosa in the antrum and body respectively.


Serum hormone levels in benign prostatic hyperplasia. The peripheral serum levels of FSH, LH, testosterone and total estrone (sum of free + conjugated estrone, mainly estrone sulphate) were determined in 20 patients with benign prostatic hyperplasia (BPH) aged 59--79 years and in 49 healthy men aged 58--79 years. There were no significant differences in the levels of FSH, LH and testosterone. The level of total estrone was significantly higher in the BPH group (p less than 0.05). The results are in accordance with our previous study on the urinary estrogen excretion in BPH and give additional support to the hypotheses concerning a role of the estrogens in the etiology of BPH.


[Prenatal alpha fetoprotein screening for defects in the neural tube]. Abnormally increased alpha1-fetoprotein (AFP) in the serum of women in the 16th--18th week of pregnancy defines a group of individuals with a high risk for open neural tube defects (spina bifida and anencephaly). Repeated measurement of AFP, ultrasound and further clinical examination serve to exclude other situations associated with high serum AFP such as threatened or missed abortion, wrongly estimated duration of pregnancy, twins, etc. Suspicion of fetal neural tube defect can be substantiated by measurement of AFP in the amniotic fluid following amniocentesis to an extent which renders interruption of pregnancy justifiable. A symposium held in Zurich is reported whose purpose was to summarize the present state of knowledge and discuss the possibility of a screening program for Switzerland. It is estimated that in this country 0.8--1.3 cases of spina bifida and 0.1--1 case of anencephaly per 1000 births occur, rendering screening justifiable. The points which prompted most discussion were ethical questions. A Swiss study group was constituted for further evaluation of the matter.


[Antibodies against various antigenic determinants in the factor VIII molecule (proceedings)]. Specific antibodies against human factor VIII have been prepared in rabbits and ducks. These antibodies differ from each other in that duck anti VIII antibody precipitates human and rabbit factor VIII, whereas rabbit antibody does not precipitate its own factor VIII. It is possible that the two antisera detect carriers differently.


Biological properties of virus-induced interferons. Both mouse and human cells elaborate more than one molecular species of interferon. Mouse interferon consists of a number of polypeptides which all probably have antiviral activity. Human fibroblast interferon is a single polypeptide, but other human cells produce special species of interferons, which show different physical and biological properties. The human interferons certainly differ in their extent of glycosylation and hydrophobicity, and may have different amino-acid sequences, since it has recently been reported that when the messenger RNAs extracted from fibroblast and lymphoblastoid cells are translated in Xenopus oocytes, the products are antigenically distinguishable (26,29,40). If the polypeptide sequences are different, then there may be at least two structural genes for human interferon. Both the antiviral and anticellular activities of mouse and human interferon probably reside in the same molecule.


Some properties of interferons induced by stimulants of T and B lymphocytes. Murine interferons induced by viruses and by non viral substances such as lectins, tilorone, lipopolysaccharide and Brucella have been analysed by affinity chromatography on a column of Sepharose coupled to anti-bodies directed against viral interferon. B cell stimulants induced interferons which are antigenically related to viral interferon and T cell dependent mitogens induced an interferon species which appears to be antigenically and physicochemically different. Despite their antigenic differences, both kinds of interferon seem to follow similar pathways for the induction of the antiviral state (sensitivity to antimetabolites, and in vitro phosphorylation of a 67 000 MW protein).


Mitral stenosis in Whipple's disease.

Mitral stenosis in Whipple's disease. A patient who died of Whipple's disease had moderate mitral stenosis with large firm yellow vegetations on the contact area of the mitral leaflets. Light microsocopy showed PAS positive macrophages within the thickened cusps and overlying vegetations. Negative images of rod-shaped bodies were visible in the cytoplasm of the histiocytes. No Aschoff bodies were seen, and there was no history of rheumatic fever. The findings in this patient lend support to the concept that chronic rheumatic type valvar deformity may result from a persistent intrinsic infectious agent.


False HLA--D assignments may be caused by cytotoxic responder lymphocytes. Lymphocytes from a healthy female repeatedly giving rise to MLC-typing response against HLA--D homozygous typing cells of three different specificities were investigated for cytotoxic capacity by the direct CML technique. Testing against a panel indicated the presence of circulating cytotoxic lymphocytes with specificity towards HLA--A2. When tested against selected HLA--D homozygous typing cells, the pattern of CML reactivity closely resembled the pattern of MLC-typing responses, i.e. typing responses were mostly restricted by the presence of HLA--A2 on the stimulator cells. This pattern was also found when time course studies of Cr--51 release were performed using experimental conditions identical to ordinary MLC typing, but involving chromium-51 labeled, irradiated homozygous typing cells as targets. These studies indicate that the presence of in vivo educated cytotoxic lymphocytes among the responder lymphocytes may in some instances mimic typing responses. Such lymphocytes are thought to lyse relevant stimulator lymphocytes prior to initiation of the proliferative response.


Low responsiveness in MLC induced by certain HLA--A antigens on the stimulator cells. An individual, BK, repeatedly gave typing responses against homozygous typing cells representing more than two HLA--D antigens. Family studies showed that she had inherited the HLA--Dw2 and Dw7 determinants, in agreement with results from primed lymphocyte typing. The HLA--A, B, C types of the stimulators giving rise to unexpected typing responses all involved the HLA--A1, A3 or A11 antigens. The hypothesis of this specificity in low-responsiveness was confirmed in the independent stimulator sample provided by the 7th workshop homozygous typing cell panel. The same pattern was observed when BK was tested as a responder towards related and unrelated heterozygous stimulators. Furthermore, in three-cell experiments it was found that BK cells were able to suppress the response to the Dw-identical individual, BS, towards stimulators carrying HLA--A1, A3, or A11. This effect of BK's cells appeared to be radiosensitive. We were not able to demonstrate suppressive supernatant factors in the relevant cultures, neither were we able to find circulating cytotoxic cells by the direct CML-technique, nor an accelerated cytotoxic response by the indirect CML-technique against targets carrying HLA--A1 or A3. This is the first demonstration of induction of suppressor cells in MLC by HLA--A, B, C antigens in the stimulator cells.


Proliferative response of H-2 incompatible leukocytes to an Abelson virus-induced lymphoma cell line apparently not expressing Ia-antigens. The proliferative response of normal lymph node cells from different mouse strains to an Abelson virus-induced leukemic cell line (MLVA) in mixed leukocyte culture (MLC) has been investigated. MLVA leukemic cells do stimulate a specific response and the stimulating determinants are controlled, most likely, by genes in the K-end of the H--2 complex. However, using two different anti-Ia sera and different assay systems, we were unable to detect the expression of Ia-antigens on MLVA. In addition, an alloantiserum raised against MLVA, and absorbed with P815, a nonstimulating mastocytoma cell line, showed no residual activity against MLVA cells.


Studies of the HLA complex in families of children with congenital heart disease. In order to test the hypothesis that HLA-linked developmental determinants are involved in the pathogenesis of congenital heart disease, we performed tissue typing on 10 families in which two or more siblings had cardiac malformations. No association between the occurrence of congenital heart disease and the HLA complex was found.


New HLA--D specificity included in Dw7.

New HLA--D specificity included in Dw7. An HLA--D homozygous typing cell -- MS -- was found to induce typing responses with a phenotype frequency of 7% tested in a panel of 202 random individuals. All but two of these were positive for HLA--Dw7, which implies that the MS cell defines a split of Dw7. The new subgroups, MS, is associated with B12. Three individuals positive for MS but lacking Dw7 were found to be B13 positive. The relationship of the MS specificity to HLA--Dw11 has yet to be defined.


HLA and Japanese MS.

HLA and Japanese MS. Determination of HLA-A, -B and -C types in 43 Japanese patients with multiple sclerosis (MS) and of DR type in 25 MS patients was carried out using antisera from the 7th International Histocompatibility Workshop. The results were compared with 46 controls typed simultaneously. Twenty-three patients were also tested for HLA-Dw2. The conclusions were: 1) There were no significantly higher occurrences of HLA-A3, B7, Dw2 or DRw2 in Japanese MS. 2) Japanese MS might nevertheless be associated with the human major histocompatibility complex, because HLA-B40 was significantly less frequent in MS and two anti HLA-DRw sera, 7w008 and 034, reacted positively more often against lymphocytes from MS patients.


Studies of HLA alloantigens of the Navajo Indians of North America. I. Variance of association between HLA-DRW (WIA) and HLA-DW specificities. As a part of the 7th International Histocompatibility Workshop, 49 randomly selected Navajo Indians were B-cell (DR) typed. Thirty-three of these individuals were also HLA-D (L.D.) typed with the Caucasian derived Copenhagen homozygous testing cell panel. DRW 2, 7, and WIA 8 were clearly defined while DRW 3 and 4 were less well defined and DRW 1, 5, and 6 were not defined in this population by the Workshop antisera. There was highly significant correlation between DRW 7/DW 7 and WIA 8/DW 8 but no correlation at all between DRW 2/DW 2, DRW 3/DW 3 and DRW 4/DW4. These results, in part, were discordant with the finding of the 7th Workshop and suggest that either DRW (WIA) and DW alloantigens are the product of more than one gene locus or that these antigens are a product of one gene locus but that each antigen molecule has multiple factors.


A conjugate of cellulase with fluorescein isothiocyanate: a specific stain for cellulose. A fluorescent technique has been developed for in situ staining of cellulose. The staining agent in conjugate of cellulase and fluorescein isothiocyanate (FITC). Application of this agent does not disturb intercellular or intracellular substances. The technique depends on the specific binding of the fluorescent labeled enzyme to its substrate. The stain has been tested on cell-free noncellulose polysaccharides similar to cellulose and does not stain them. The technique has been used to localize cellulose during the life cycle of Dictyostelium discoideum with results that correspond to previous work using other methods.


Effects of hydrogen ion dissociation and concentration of the reactivity of dichromate-fixed tissue components to the PAS procedure: recommendations for reducing undesirable background staining. The undesirable PAS reactivity of cytoplasmic aldehydes after dichromate fixation can be suppressed without affecting selective staining by lowering the pH of Lillie's Cold Schiff reagent to 1.5. Alternatively, dilution of pH 2.2 Cold Schiff reagent with distilled water (1:2) is recommended. Hydrogen ion concentration and dissociation effect the rate of color formation in various PAS positive sites differentially with respect to the time of incubation in Schiff reagent. Based on the experiments, aldehydes exposed in different tissue components appear to be chemically distinct and separable depending on the rate of color formation and duration of incubation in Schiff reagent.


Identification of oleoresin in epoxy-embedded slash pine tissue. Sudan black B stains oleoresin blue-black in epoxy-embedded material as well as in living tissue. The Sudan black B staining properties of oleoresin are similar to those of lipid, but it can be distinguished from tannin, which stains brown. Pratically all oleoresin present in resin  ducts and intercellular spaces, and much of that contained in epithelial and ray cells, is extracted in preparatory procedure for electron microscopy. A fixation procedure is proposed which preserves significantly more oleoresin in situ. The use of Sudan black B enables one to localize oleoresin by light microscopy, and permits direct comparison of adjacent sections of epoxy-embedded material at the ultrastructure level. Ultrastructurally oleoresin and lipid possess similar electron densities and can be distinguished from the highly electron-opaque tannin deposits.


Improved nuclear staining with mordant blue 3 as a hematoxylin substitute. For progressive staining 1 g mordant blue 3, 0.5 g iron a alum and 10 ml hydrochloric acid are combined to make 1 liter with distlled water. Paraffin sections are stained 5 minutes blued in 0.5% sodium acetate for 30 seconds and counterstained with eosin. For regressive staining, 1 g dye, 9 g iron alum and 50 ml acetic acid are combined to make 1 liter with distilled water. Staining time is 5 minutes followed by differentiation in 1% acid alcohol and blueing in 0.5% sodium acetate. Counterstain with eosin. In both cases results very closely results very resemble a good hematoxylin and eosin.


Enhancing aniline blue fluorescent staining of cell wall structures. Prior staining with the periodic acid-Schiff reaction, toluidine blue O, Congo red, or Calcofluor White M2R New, or reduction by NaBH4 do not interfere with aniline blue-induced fluorescence of sieve plates, new cell walls, pit fields or tracheids in compression wood of conifers. Detail of such fluorescent structures is improved by these treatments because of increased contrast, reduced flare, and a quenching of autofluorescence.


Simple and durable staining of thick sections of the human brain for macroscopic study. Observations on gross morphology of the brain is greatly facilitated by enhancing the contrast between gray and white matter. The proposed technique is much more simple than the generally recommended Mulligan method and it variations. Moreover, there is no loss of stain since the fugitive surface impregnation, obtained by the Mulligan method, is replaced by a thoroughgoing block-staining procedure with the nonfading copper phathalocyanine dye astra blue. Staining procedure: wash formalin-fixed brain slices overnight in running tap water. Place slices in performic acid for 1 hour. Wash in running tap water. Place slices individually in staining solution consisting of 0.1 g astra blue (Merck) in 1000 cc distilled water and 1 cc HCl (37%), for 12-24 hours. Wash in running tap water. Embed in gelatin and mount in plastic cuvettes.


Methods of preparation of fossil bone samples for light and transmission electron microscopy. The methods described make possible the preparation of fossil samples for light nad transmission electron microscopy. To obtain good images with ground sections of bone, both the grinding process and classical bone staining techniques were modified, the latter by staining already made ground sections. Ground sections of suitable thickness were etched with a 4:1 mixture of 5% HNO3 and 2% OsO4 at a lowered temperature, thereby obtaining isolated osteocytes with a complete network of processes. Using the same method with thicker ground sections, the contents of vascular canals were isolated. The contents retained spatial relations permitting complete reconstruction of the vascular system of the bone in question. The isolated osteocytes, the vascular canal contents, and the isolated remnants of the intercellular organic part of this bone, subsequently embedded in Epon 812 and in methacrylate, made possible sections suitable for detailed analysis of the image of the surfaces of these elements. Polished surfaces of the fossil bone were replicated by the so-called clearing replicas technique adapted to fossil bone. The latter facilitated analysis of the ultrastructure of the surfaces of the vascular canals, isolated osteocytes and their processes, and analysis of the immediate proximity of these elements, since the technique preserves the mutual spatial relations intact.


[Efficiency of radiation therapy in the treatment of epipharyngeal tumors (author's transl)]. The radiotherapeutic results of 38 patients with malignant tumours of the epipharynx are analyzed. Only five patients had T1 and T2 lesions, while all the others suffered from more advanced tumours. Five patients revealed distant metastases at the beginning of palliative radiotherapy. For this reason, the 5-year survival rate was only 16%. Conversely, a local tumour control was achieved in more than two thirds of the patients (27/38). This result gives support to the suggestion that radiotherapy, either in combination with surgery or without it, is to be considered at this time as the most effective method of treatment for malignant tumours of the epipharynx.


[Fluorescence anisotropy and myelin structure. II. Azimuth characteristic of surviving nerve fibers]. The dependence of fluorescence polarization of stained nerve fibres on the angle between the fibre axis and electrical vector of exciting light (azimuth characteristics) has been considered. Evidence is provided that the azimuth characteristics of stained nerve fibres depends on dye molecules adsorbed on the myelin sheath membranes. From the previous calculations it may be concluded that part of the dye molecules are oriented at a small angle to the geometrical axis of the nerve fibre.


[Fluorescence anisotropy and myelin structure. III. Effect of temperature changes on the orientation of dye molecules in nerve fibers]. The temperature dependence of fluorescence polarization of stained nerve fibres has been studied. As has been previously demonstrated by the authors, the dependence of fluorescence polarization on the angle between the electrical vector of exciting light and the fibre axis (azimuth characteristics) is associated with the molecular orientation of dyes adsorbed on the membranes of the myelin sheath. This permits an indirect conclusion to be made about the structure and structural changes of an adsorbent. The experiments with changing temperature show that the molecular orientation of dyes decreases with decline of temperature from the room temperature to the freezing point of the Ringer solution. The structure of myelin membranes is suggested to be stabilized through hydrophobic interaction.


[Immunochemical characteristics of the peroxidases of several mouse tissues]. Enzymes catalyzing peroxidase reaction of a lysosomal fraction in bone marrow, leucocytes, spleen, thyroid gland, stomach, kidney, heart, lungs, brain and skeletal muscle of mice were investigated by immunochemical methods. A high level of peroxidase activity was discovered in leucocytes, bone marrow, spleen, heart and lung, a lower activity appeared to be characteristic of liver, thyroid gland and kidney. The peroxidase activities in brain, skeletal muscle and stomach were low. The reaction of immunoprecipitation with myeloperoxidase-specific antiserum revealed considerable antigenic distinctions between the enzymes catalysing peroxidase reaction in various tissues of mice.


Prenatal diagnosis of neural tube defects through amniotic fluid measurements. On the basis of our experience with 1314 prospective diagnoses for neural tube defects, we continue to rely on amniotic fluid AFD as the primary and most reliable indicator. The problem of fetal blood contamination has to be tackled in a quantitative manner, and an attempt made to estimate the likely contribution from fetal serum based on measurements of the proportion of fetal erythrocytes in the mixture. We also recommend estimation of the macrophage proportion at 18 hours in the cultured amniotic fluid cells, and a detailed examination of the morphology of cells that adhere, at this time, to the culture surface. An outline of the procedure that we currently employ in handling amniocentesis samples is shown in Figure 3.


The importance of accurate gestational age estimation in screening for fetal neural tube defects using maternal serum alpha-fetoprotein levels. Estimates of gestational age based on menstrual history are often in error, often by as much as 4 weeks. The effect of this problem with respect to a screening program for neural tube defects based on maternal serum alpha-fetoprotein determination is examined. The variance of alpha-fetoprotein values for serum samples assigned to gestational age categories from 15 to 19 weeks of gestation based on menstrual history and that of samples classified according to sonographic estimates do not show clear-cut differences. However, when an individual sample is referred to a calibration curve of normal values, errors in gestational age estimates significantly increase the possibility of a false positive/false negative decision for high-level percentile classification. The effect of such an error is least at 16 to 17 weeks of gestation.


Pitfalls and problems in the interpretation of AFP data--myths and misuse of the "normal range". Drawing a line more intelligently for AFP data means using as a denominator two fairly well-defined populations (abnormal and normal), instead of one ill-defined normal population. The line is drawn at the intersect between two curves to minimize the error of misclassification; this replaces an arbitrarily defined cutoff level (multiples of the standard deviation unit). Moving the line to the right reduces the false positive error when screening for NTD, whereas moving the line to the left provides a sensitive alert signal for routine screening of maternal sera by AFP determinations for fetal distress. Adjustment of the line for cost and benefit factors and the use of probability statements that include the prevalence of specific conditions all help sharpen medical classification and decisions. Cross-sectional analysis is separated from longitudinal analysis. A benefit from this discussion may be the replacement of the normal range with a simple plot of AFP results or a percentile distribution, which would be more meaningful and more easily understood by all.


Biologic tumor markers in management of testicular and bladder cancer. The role of biologic markers in the management of testicular and bladder cancer has been described. alpha--fetoprotein (AFP) and human chorionic gonadotropin when used together have been extremely helpful in detecting recurrence, staging, and monitoring of testicular cancer. The cellular localization of these markers by immunoperoxidase has also been an aid in understanding the natural history of these tumors. The cell surface antigens in bladder cancer have been useful predictors as to whether or not Stage A bladder cancer will develop into more invasive cancer when compared with the conventional histopathologic examination of the bladder tumor. In the last six years we have prospectively utilized these markers in the management of testicular and bladder cancer. The clinical and laboratory findings of these prospective studies are reviewed.


Detrusor pressure in bladder outlet obstruction. Synchronous urinary flow and pressure studies were done on 51 male adults consiting of 12 normal subjects and 39 patients with varying degrees of bladder outlet obstruction due to benign prostatic hypertrophy. Mean maximum detrusor pressure levels of 50 mm. Hg and above were found to be indicative of bladder outlet obstruction. Elevated mean maximum detrusor pressure fell preoperatively within normal limits after prostatecomy. Chronic retention of urine was not associated with poor contraction of the detrusor muscle. Clinical findings of obstruction were shown to be inadequate for estimation of maximum detrusor pressure. Recording the detrusor pressure was found to be a better urodynamic test for bladder outlet obstruction than recording the urinary flow rate. Recording urinary flow is, however, a simple test and should remain a screening test for impaired drainage of the bladder outlet.


Effect of periodate oxidation on specific activities and carbohydrate components of human blood group N- and M-specific glycoproteins and glycopeptides. Mild as well as strong periodate oxidation of isolated erythrocyte N and M glycoproteins and glycopeptides gave extensive to complete destruction of N-specificities as measured with Vicia graminea extracts and of N- as well as M- activities determined with all but 1 of 8 animal anti-N and 13 anti-M sera. Results with human antisera differed somewhat, while the specificity of mildly oxidized N-glycoprotein was completely destroyed as determined with all 8 human anti-N sera used, that of strongly oxidized N-active substance was completely inactivated towards one of the human antisera; the remainder showed 63--94% destruction, and 2 sera indicated no effect of oxidation. Similarly, while 10 of 14 human anti-M indicated complete inactivation of M-specific glycoproteins and glycopeptides after mild or strong oxidation, 2 showed partial inactivation and 2 human anti-M sera showed no inactivating effect of oxidation. The most relevant findings of quantitative carbohydrate analysis of periodate oxidized N- and M-specific substance were extensive transformation of N-acetylneuraminic acid (NAN) to its C8 and C1 analogues on mild oxidation and pronounced destruction of NAN and its analogues on strong oxidation; however, some intact NAN always remained. In all instances galactose (Gal) was destroyed to a much larger extent in N-derived than in M-derived glycoproteins and glycopeptides.


New hepatitis B surface antigen subtypes inside the ad category. In addition to the 10 HBs Ag subtypes already described, 2 new subtypes were defined by using the q determinant. Exceptions to the rule generally accepted were found in that the q determinant was only lacking in HBs Ag/adw4. These exceptions occurred in adw and adr categories. These 2 new subtypes are adw q positive and adr q negative. Out of 98 HBs Ag/adw4 from silent carries and patients from different parts of the world, mainly from France (79), 6 were found q positive. 3 out of these 6 cases came from Montpellier (South of France) and another 3 from Germany. The 92 other cases were found q negative. Further studies will be necessary to better know the location of this new subtype adw4 q positive, but it seems to be present only in certain parts of Europe. Out of 86 HBs Ag/adr from silent carriers from Asia (58), Oceania (17), France (10, most of them contaminated in Asia) and Réunion (1), 10 were found q negative. All these 10 cases were detected in Oceania, 2 out of 2 in carriers from New Caledonia and 8 out of 13 from French Polynesia. The new subtype adr q negative seems localized in Melanesia and Polynesia and absent from Asia. These 2 new markers of hepatitis B virus will allow better epidemiological and geographical studies.