Phantom urinary bladder pain--case report.

Phantom urinary phenomena are a relatively rare disease entity. Literature search has revealed only one case following cystectomy, seven cases following spinal cord injury and several other cases in hemodialysis patients. This report presents a case of painful phantom bladder following cystectomy for chronic kidney and urinary tract infection. Treatment was directed toward sensory hyperstimulation for suppression of the subjective experience of the painful phantom. Lumbar sympathetic blocks and transcutaneous electrical stimulation were used. Competent coping mechanisms were increased through relaxation training and assertiveness training to deal with a medical problem which has no standard solution. The patient's response to the Comprehensive Pain Control Program was excellent with an estimated 75% reduction in painful phantom perceptions. A brief discussion of the medical literature on the subject is presented.


A portable electronic 'calling device' as an aid to 'weaning' ventilator-dependent tetraplegic patients from intermittent positive pressure ventilation.

An electronic calling device has been developed. It is easily portable and can be operated by the severely paralysed tetraplegic patient in order to attract attention. The patient can operate the sensor either during artificial ventilation or when breathing spontaneously via the tracheostomy tube at a stage prior to the insertion of a speaking tube.


A new pressure measuring system for cushins and beds--with a review of the literature.

An interface pressure measuring device for the assessment of subject-cushion interface pressures at ten points has been described. Preliminary trials using 12 healthy subjects on four cushions and three spinal injury patients on six cushings, has been carried out. It was also found that the water and foam cushion distributed the pressure over a large area than the foam cushions.


Differences in virulence of Naegleria fowleri.

All pathogenic Naegleria fowleri isolated from the environment were highly virulent to mice when instilled intranasally. Axenic cultivation gradually decreased virulence of highly virulent strains. This decrease was most pronounced in environmental isolates and of minor importance in N. fowleri isolated from human cerebrospinal fluid. The low virulent strains obtained by continuous axenic cultivation appeared after clonation to consist of individuals with different virulence. Virulence could be enhanced in low virulent strains by brain passage and passages in Vero cell cultures, but could not be induced by these methods in nonvirulent strains isolated from the environment. Different mice strains showed different sensitivities to infection with pathogenic Naegleria. In addition, older mice were less sensitive than younger animals to low virulent strains.


[Incidences of resistance capacity (RC) and of electrode-interval enlargement on the gastro-intestinal electrical activity of the awake dog (author's transl)].

By the use of differents resistance capacity (RC) values (0,3, 0,7, 1, 3, 6 and 7 s.) during bipolar recordings of digestive electromyogramme in chronically implanted dogs, it has been shown that the RC incidences were:--an increase of the slow wave potential amplitude, the optimale value of the RC was 3 sec;--the detection with long RC (1, 3, 6 and 7 S.) of a slow post-potential at the antral level. But there is no effect of the RC on the slow wave duration and on the spikes potentials caracteristics (amplitude-duration number). When the interval between the two active electrodes is increased (0,5 to 63 cm) the slow wave amplitude is enlarged; the best results on the slow wave amplitude are obtained with an electrode-interval of 7 to 8 cm. In cases of an important interval between two active electrodes (8, 16, 32, 63, 190 cm), the slow wave frequency at an intestinal level is dependent of the upward electrode. The spikes potentials are affected by electrode interval enlargement, and are dependent of the contractil events which can appear under each separated electrode.


[Plasma cell beta-glucuronidase and monoclonal gammapathies (author's transl)].

beta-glucuronidase activity, estimated cytochemically, is, in more than half cases of myeloma, very increased or decreased, compared to that found in benign dysglobulinemias, where this activity is normal. A very high or very low score makes sure the diagnostic of myeloma. The relations between the beta-glucuronidase activity and the myeloma clinical stade according to Durie and Salmon and the bone lysis are discussed.


[Thrombopenia and radial aplasia: 2 cases with platelet function and ultrastructural studies of megakaryocytes and platelets (author's transl)].

The authors report on two cases of congenital thrombopenia with radial aplasia. Both children display several formative abnormalities and a mild thrombopenia; hemorragic manifestations occurred in the first case only. Megacryoblastic to platelets series, as studied with electronic microscopy, show small-sized, "microcytic" and hypogranular megacaryocytes, displaying a maturative disorder (dysmegacaryocytopoiesis). In functional studies, platelets of the first patient show an imperfect nucleotidic release and do not agregate normally with ristocetin. The second case exhibits mostly a PF3 reduction. The variety of expression of the megacaryocytic-platelets disorders appears likewise in the squelettal and visceral malformations. The whole disorder could be ascribed to a pleiotropic abnormal gene with a variable expressivity.


[Enzymatic induction during isoniazid therapy (author's transl)].

Enzyme induction by isoniazid was studied by urinary D-glucaric acid estimation in slow and fast acetylators. Isoniazid administration increases significantively the D-glucaric acid elimination in the two classes of patients. In fast acetylators, the glucaricaciduria increases regularly up to 1.8 fold the physiological level in 30 days. In slow acetylators, after a progressive elevation during 20 days, the glucaricaciduria reaches quickly a 3 fold increase after 30 days. When a classic inducer such as phenobarbital is administrated in association with isoniazid, induction is stimulated in the two groups of patients. The estimation of plasmatic free isoniazid seems to indicate that the acetylation rate of isoniazid is not on the dependance of the induction process when the drug is administrated alone. In contrast, this rate increases when the association isoniazid-phenobarbital is administrated to slow acetylators. Possible consequences on the hepatic toxicity of isoniazid are discussed.


[Use of serum s-IgA detection in liver pathology (author's transl)].

Secretory IgA (sIgA) were searched in 60 sera of healthy blood donors and in 1 590 sera of subjects having various diseases. 20 percent of these subjects showed an increased amount of sIgA in their sera. The only subjects presenting a constant increase (sometimes more than 20 fold the normal amount) were people with liver diseases. Quantitation of sIgA, in relation with the determination of the IgA/transferrin ratio (IgA/T) in sera, showed an important difference between Laennec's cirrhosis on one hand and virus hepatitis or post-hepatitic cirrhosis on the other. In Laennec's cirrhosis a moderate increase in sIgA went with a strong elevation of the IgA/T ratio, the latter being proportional to the degree of evolution of the disease. In virus hepatitis, the sIgA amount was largely increased while the IgA/T ratio remained at a normal value.


[Detection of blood group P1 activity in F. hepatica and F. gigantica (author's transl)].

Four lyophilizated liver flüke antigens and one of hydatid fluid were examined for the presence of blood group like antigens P1, A, B, M and N by an hemagglutination inhibition test with hyperimmune and natural hemagglutinating sera. P1 or P1-like substance was found in the extracts of "Fasciola hepatica" and "F. gigantica" in a lower concentration than in echinoccocal fluid. This presence may explain the occurrence of anti-P1 antibodies in patients infected with liver flüke and belonging to the P2 blood group. However biochemical studies will be necessary to determine if the P1 activity is due to the blood group P1 substance of the red cell itself or due to a P1 like substance.


[Influence of reintervention by the bacillus of Calmette and Guérin (BCG) on the nonspecific resistance of the mouse to Schistosoma mansoni (author's transl)].

Significant resistance to Schistosoma mansoni appears in mice two weeks after treatment with BCG. The state of nonspecific resistance is considerably increased if the injection of BCG two weeks prior to the infestation test is carried out using mice who have already received BCG. Thus, BCG innoculation in an organism presentized to the bacillus antigens increases immunostimulation. According to these results, during primoinnoculation with BCG, immunostimulation is not significant before this second week, that is, the time necessary for a state of hypersensitivity to appear. Nevertheless, it should be noted that weak yet detectable immunostimulation can be observed before the 14th day following a BCG primoinfection. Thus, the possible role of other mechanisms should be considered.


The survival of cryopreserved human bone marrow stem cells.

Two methods for cryopreservation of bone marrow stem cells were compared using bone marrow obtained from 36 patients. Included in this group were 21 persons with the diagnosis of leukaemia including 14 either with acute myeloid or lymphoblastic leukaemia in remission following intensive remission induction chemotherapy. After freeze-preservation and reconstitution, all marrow samples were tested for nucleated cell (NC) recovery and grown on agar to assess colony forming units (CFUC) and cluster forming units in culture (CluFUc). A slow dilution reconstitution method using freezing media containing AB negative plasma resulted in recovery of 85% of the CFUc activity of fresh marrow. This result was significantly better than the 47% CFUc recovery obtained when freezing media without plasma and a rapid dilution reconstitution technique were used. NC recoveries following slow dilution (51%) and rapid dilution (44%) were not significantly different. CluFUc were disproportionately reduced compared with CFUc although yielding similar results with both methods (26% and 32%). No correlation was found for either method between CFUc and NC recovery or between CFUc and CluFUc recovery in cryopreserved bone marrow.


A solid-phase enzyme-linked immunosorbent assay (ELISA) for detection of antibody to rubella virus.

An enzyme-linked immunosorbent assay was established for detection of antibodies to rubella virus. In this system commercially available rubella antigen was attached to the wells of polystyrene microtitre plates after which sera were added and incubated to allow the formation of antigen-antibody complexes. The presence of bound antibody was detected by adding anti-human globulin coupled to horseradish peroxidase and visually observing the colour change produced after addition of an appropriate substrate. The test was reproducible and simple to perform and had a similar sensitivity to the widely used haemagglutination inhibition system.


Surface marker studies in chronic lymphocytic leukaemia and non-Hodgkin's lymphoma.

Immunological surface marker techniques were applied in a study of 29 cases of chronic lymphocytic leumaemia and 22 of non-Hodgkin's lymphoma. Surface marker characteristics distinguished 2 subtypes of B lymphocytes. Chronic lymphocytic leukaemia was a monoclonal proliferation of B lymphocytes which produced spontaneous rosettes with mouse erythrocytes and had faintly immunofluorescent surface immunoglobulin. The majority of non-Hodgkin's lymphomas also had their origin from B lymphocytes but in contrast, this subtype did not show receptors for mouse erythrocytes and their surface immunoglobulin was brightly staining and demonstrated "capping". The clonal origin of nodular lymphomas could also be demonstrated on frozen sections stained for surface immunoglobulin. Two cases of true histiocytic lymphoma were identified. The current information available on surface marker characteristics of the leukaemias and lymphomas is reviewed.


Transplacental passage of islet cell antibody in infants of diabetic mothers.

The incidence of pancreatic islet cell antibody (ICAb) was assessed in the cord blood and sera of infants of diabetic mothers (IDM). ICAb activity was detected in the cord blood of 10 of 36 (27.8%) consecutive IDM studied and in 0 of 111 (0%) cord bloods of normal control infants. In all instances, ICAb activity in the cord sera was of the immunoglobulin (IgG) class and was associated with ICAb in maternal sera at the time of delivery. No correlation was observed between the incidence of ICAb in IDM and alterations in fetal growth parameters, congenital malformation rates, cord blood insulin levels, or the incidence of neonatal complications. The evidence would support transplacental passage of ICAb from diabetic mothers to their offspring, but would not support a primary pathogenetic role for ICAb in the clinical or metabolic alterations observed in these infants.


Deficient alternative complement pathway activity in newborn sera.

Neonatal susceptibility to overwhelming bacterial infection is commonly attributed to a relative deficiency in serum opsonic activity. However, few studies have compared the functional capacity of the classical complement pathway with that of the alternative complement pathway in the neonate. The opsonic activity of nine maternal infant serum pairs were studied by determining percent uptake of radiolabeled Escherichia coli. Seven mother-infant paired sera were studied using E. coli strains known to be opsonized via the alternative complement pathway: the mean percent uptake of E. coli opsonized in neonatal sera was 16.8%; of those opsonized in maternal sera, 54%; and of those opsonized in control sera, 45% (P less than 0.005). Two E. coli strains requiring the classical complement pathway for opsonization were phagocytized equally well in maternal and infant sera of seven mother-infant pairs. Determination of anti-O hemagglutination inhibition (HI) antibody titers in six maternal sera for one classical complement pathway activating and one alternative complement pathway strain showed no correlation between percent phagocytosis and HI antibody titer. These data would suggest that serum levels of classical pathway components are probably adequate for opsonization of E. coli via the classical pathway, but that low alternative complement pathway activity in neonatal sera may contribute to the newborn's increased susceptibility to bacterial sepsis.


Haemophilus parahaemolyticus serotypes. Pathogenicity and cross immunity.

Pigs inoculated intranasally with Haemophilus parahaemolyticus, serotype 2, resisted challenge 3 weeks later with serotypes 2, 4 and 5 without showing clinical symptoms. The pigs were sacrificed 2 days after challenge, and post mortem examination showed a chronic pleuropneumonia from which only serotype 2 was re-isolated. Pigs inoculated intranasally with H. parahaemolyticus, serotype 2, showed no clinical symptoms when challenged 3 weeks later with serotype 1. Post mortem examination revealed a chronic pleuropneumonia with areas of necrosis from which H. parahaemolyticus, serotype 2, was re-isolated, but also small areas of a more acute fibrinous pneumonia from which serotype 1 was re-isolated. The control pig inoculated with only serotype 1 showed a severe acute fibrinous pleuropneumonia. The results indicate that a considerable cross immunity exists between the various serotypes of H. parahaemolyticus.


The ulcus-syndrome in cod (Gadus morhua). III. A preliminary virological report.

A bullet shaped virus presumably belonging to the Rhabdoviridae was isolated from a pool of 7 cods (Gadus morhua) with the ulcus-syndrome. This virus measured 55 x 175 nm. The virus caused CPE in PS (pike sarcoma) cells, but not in a number of other cell lines. From the same pool another virus was isolated in EPC (epithelia carp) cells. This virus was an icosahedron of 145 nm and most likely to be placed in the family Iridoviridae. Electronmicroscopic examination of papules from another 13 cods with the ulcus-syndrome failed to reveal any virus. Attempts to isolate virus from 33 individual cases of the ulcus-syndrome were successful in one single case only. The two viruses have not consistently been isolated from cods with the ulcus symdrome, but on the other hand they have never been isolated from cods without the ulcus-syndrome.


Influence of storage and preservation of milk samples on microscopic and Fossomatic somatic cell counts.

Milk somatic cell counting was carried out by Fossomatic and microscopically. In unpreserved milk samples Fossomatic counts increased by up to 100% during the first 24 hours after milking. During the next 24 hours the counts increased by 5% whereafter they remained stable until at least 80 hours after milking, when the samples were stored at 5 degress C. On microscopical counting using methylene blue for staining, the results were stable from shortly after milking. In counting, attention had to be paid to the fact that within the first 24 hours a certain part of the cells would stain but faintly. After preservation with potassium bichromate the Fossomatic counts increased more rapidly. Stable results were found 5--8 hours after preservation. The final level of the Fossomatic counts was found to be app. 10% higher in preserved than in unpreserved samples. A smaller increase was found by microscopic counting. The rise of the cell counts during the first 24 hours after milking is probably due to inadequate stainability of living cells with ethidium bromide, resulting in a certain part of them being recorded by the Fossomatic. During the first day the vitality of the cells diminishes whereby they become stainable and countable. This process in hastened by potassium bichromate treatment.


[The surgical treatment of large incisional hernias by lacing of autogenous full thickness skin. 36 cases (author's transl)].

Strips of skin taken from the excess skin adjoining the scar, prepared carefully and used as a loose continuous suture, represent an elastic material which is easy to use and of great strength. Results studied in a group of 36 patients with a follow-up varying from 12 month to 16 years were excellent in 70% of cases and moderate in 25%. The quality of the results obtained, from both a mechanical standpoint as well as in terms of tolerance, together with the simplicity of the technique, have led us to consider it as a method of choice in the repair of large incisional hernias, in particular recurrent.


[Listeriosis in the kidney transplant recipients. Ten cases (author's transl)].

Three clinical pictures were found in ten cases of listeriosis after renal transplantation: septicemia (six cases), meningo-encephalitis (three cases) and asymtomatic but recurrent bacteriuria (one case). Several features demonstrate profound immunologic disturbances in the recipient: close relationship in 9 cases out of 10 between infection and high doses of immunosuppressive drugs, unfavorable outcome of graft (accelerated rejection in 4 cases out of 10) and association in one case with lethal pulmonary infection due to opportunistic agents (cytomegalovirus and Pneumocystis carinii).


[Initiation in vivo of blood coagulation. The role of white blood cells and tissue factor (author's transl)].

Tissue factor is an ubiquitous phospholipid-protein complex, which triggers blood coagulation through the so-called extrinsic pathway. Reactions initiated by tissue factor bypass many of the early stages of coagulation (contact phase) and involve factors VII, X, V, II and fibrinogen but also factor IX (and VIII) as it was recently demonstrated. So, it appears that tissue factor has a key-role in the haemostasic process as it has been suggested by the mildness or the absence of haemorrhagic syndrome in contact factors deficiencies. Tissue factor activity has been found in many types of cells, especially in white bloods cells. Experimental studies have demonstrated the presence of tissue factor activity in polymorphonuclears, lymphocytes, monocytes (or macrophages). This activity is enhanced by gram-negative endotoxin stimulation, inflammation, cell mediated immunologic phenomena or malignancy. These data are in good agreement with a wild range of features observed in human pathology: fibrin deposits in inflammatory lesions, disseminated intravascular coagulation (DIC) during the course of gram-negative septicemias or acute promyelocytic leukemias, local thrombi at the early phase of graft rejection. The protective effect of a phospholipase C against DIC induced in rats by tissue factor infusion suggests in the future, a specific therapy would be possible in man that, in the frequent clinical conditions involving clotting activation through tissue factor pathway.


[Transanal excision of villous tumours. Value of the GIA automatic suture clamp (author's transl)].

The GIA (gastro-intestinal anastomosis) suture clamp facilitates hemostasis during transanal excision of villous tumours, notably when they are situated 8 to 10 cm from the anal opening.


[Rotation flaps of the leg in osteitis (author's transl)].

The authors describe a technique of rotation of a leg flap which is divided into several phases : the flap is prepared and left in place, then two or three weeks later is applied to the area of loss of substance. Finally, the donor site is grafted. This unusual technique gives excellent results when used to cover loss of substance secondary to cutaneo-osseous sepsis of the leg. This was the case in 5 of our patients treated in this way.


[Disseminated sclerosis: diagnostic value of neurophysiological investigations (author's transl)].

Demyelinisation lesions of disseminated sclerosis actually present within the white matter are much more numerous than clinical manifestations would suggest that the majority of these lesions are clinically latent. Ophtalmologists were the first to seek a functional deficit reflecting the demyelinisation which so often affects the optic pathways. In practice, the recording of evoked visual potentials has proved to be the most reliable method. The brain stem is another region where demyelinisation lesions are frequent and quite often latent. Electronystagmography offers a highly sensitive means for the investigation of internuclear connections, whilst other authors use evoked auditory potentials. Similarly, evoked somaesthesic potentials make it possible to reveal lesions of the lemniscal tract.


Structure of the promoter region for the rrnB gene in Escherichia coli.

The nucleotide sequence of the promoter region for the rrnB gene of E. coli had been determined by the Maxam-Gilbert technique. The 700 bp long sequence had been compared with the published sequences of four other rRNA promoter regions. The rrnB sequence was found to be homologous with the rrnA promoter sequence till the 370th base upstream from the coding region of mature 16S rRNA. The significance of this homology is discussed and a tentative model is proposed to account for the unusual properties of the rRNA promoters.


The nucleotide sequence preceding and including the beginning of the ilvE gene of the ilvGEDA operon of Escherichia coli K12.

The DNA sequence of the 570 base pairs that precedes and includes the beginning of the ilvE gene shows no evidence of a leader-attenuator region. Instead, the sequence shows that the ilvE gene is preceded by another structural gene, presumably the normally cryptic ilvG gene. In vitro transcription of the two plasmids (pLC26-3 and pRL5) containing the regulatory region of the ilvEDA operon results in the formation of two "leader" RNAs. These results are consistent with the suggestion that the ilvEDA operon is regulated by an attenuator mechanism and that the attenuator region lies before the ilvG gene.


The effects of mercury-substitution on the hybridisation characteristics of nucleic acids.

The effect of different levels of mercury substitution on the rate and extent of hybridisation of globin mRNA with a complementary DNA (cDNA) copy has been investigated. It was found that mercuration significantly reduces both the rate of hybridisation and the extent of the reaction, but that these effects are abolished when at least stoichiometric amounts of 2-mercaptoethanol are included in the hybridisation medium. As a preliminary to using this technique to isolate specific groups of sequences after long-term hybridisations, we have investigated both the rate of demercuration of RNA and its retention on thiol-sepharose columns after extended incubation under commonly employed hybridisation conditions at 43 degrees or 60 degrees. Retention was essentially quantitative even after incubation times of 300 hours at 43 degrees, but decreased significantly after 48 hours at 60 degrees. It is concluded that thiol-sepharose chromatography offers considerable advantages over hydroxyapatite chromatography for the recovery of hybridised sequences, particularly with regard to the lower levels of non-specific binding obtained and its ability to distinguish directly between DNA-DNA and DNA-Hg RNA hybrids.


The 3' terminus of 16S rRNA: secondary structure and interaction with ribosomal protein S1.

We report studies of the secondary structure and S1 ribosomal protein binding properties of the colicin fragment, containing 49 residues from the 3' terminus of E. coli 16S rRNA. Temperature jump relaxation kinetic measurements reveal two helices in the structure. One of these, melting at 81 degrees C in 5 mM Mg2+, is associated with the 9-base pair hairpin helix predicted by the nucleotide sequence. The other melting transition, at 21 degrees C in 5 mM Mg2+, is assigned to a 4-base pair helix which constrains the pyrimidine tract of the colicin fragment into a bulge loop. S1 protein forms a strong 1:1 complex with the colicin fragment, with an association constant of 5 x 10(6) M-1 in 5 mM Mg2+. More protein molecules are bound, but with weaker affinity, when the S1 concentration is increased. S1 binding causes melting of the colicin fragment secondary structure, as inferred from the observed absorbance increase. The S1 binding site on the colicin fragment has been localized in the region of the bulge loop, since the melting transition corresponding to the 4-base pair helix is lost in the complex. We discuss current models for the role of S1 protein in polypeptide chain initiation in light of these and previous results.


An improved rapid enzymatic method of RNA sequencing using chemical modification.

A version of rapid gel sequencing procedure based on the analysis of partial endonuclease hydrolizates of chemically modified 5'-32P-labelled RNA is suggested. Complete and selective modification of cytidilic residues by a methoxyamine-bisulfite mixture leads to the unfolding of the RNA secondary structure and, due to this effect, to the generation of a more uniform set of fragments after partial RNAase hydrolysis. The position of cytidines in an RNA sequence can be determined by restricting the hydrolysis of phosphodiester bonds between the modified CMP residues and their 3'-neighbours with T2 and A RNAases. The method was verified with tRNATrp (yeast) and 5S RNA (rat liver and yeast).


Antifungal therapy, 1978.

The past 40 years have brought great advances in the knowledge of morphology, immunology and epidemiology of fungal infections in man. Each general advance in the science of medicine has had its counterpart in man's improved facility to deal with these infections therapeutically. Although satisfactory control of fungal infections of man is still lacking, productive paths of investigation seem to have been found and the continued application of rigorous discipline to research efforts should provide even more efficient treatment in the future.


Dermatophyte infections.

The newer antifungal agents, clotrimazole, miconazole and haloprogin are considered for their efficacy and acceptability, and are compared with other topical agents used for the treatment of dermatophyte infections of the skin.


Changes in conditioned performance and general behavior produced by imipramine treatment in dogs.

Dogs with electrolytic damage of dorsomedial amygdala and lateral hypothalamus manifested the syndrome of depression. They were used as models for studying the effects of a tricyclic antidepressant. Imipramine treatment produced increase in general arousal, motility and reactivity as tested by various tests, changes in social behavior and only slight improvement of instrumental conditioned performance. In normal dogs the imipramine administration in most subjects resulted in either no effect or deterioration of various learned tasks. Strong individual differences in the reactivity to drug treatment were observed in both normal and lesioned (depressed) dogs.


Chronic mucocutaneous candidiasis.

The pathogenesis of chronic mucocutaneous candidiasis is surveyed. Treatment comprises topical antifungal treatment which is insufficient, systemic antifungal treatment which is often followed by a rapid relapse, and specific immunotherapy with live tissue or transfer factor. Combination of systemic antifungal therapy and immunotherapy seems to be the most promising approach. However, no permanent cure has so far been achieved.


Aspergillus fumigatus and lung disease.

The range of clinical presentations of lung diseases associated with Aspergillus spp. is great. The conditions are very frequently misdiagnosed but errors should be avoided if the possibility of a fungal cause is considered and simple immunological tests undertaken. Often no more than skin-prick tests and serum precipitins need to be done. In many cases, fungus is not isolated from the sputum and negative results do not exclude the possibility of A. fumigatus as the causal agent. Treatment often results in marked clinical improvement and there is evidence that suppression of recurrent episodes of bronchopulmonary aspergillosis prevents progressive lung damage.


Evaluation of vaginal antifungal formulations in vivo.

Relatively simple and rapid procedures have been developed for evaluating the local efficacy of vaginal antifungal agents in vivo in a vaginal candidiasis model in ovariectomized rats. The results of this investigation indicate that the model and methods described are quite suitable for screening potential antifungal substances and for assessing the chemotherapeutic effectiveness of new antifungal agents and formulations before carrying out clinical studies.


The polyene macrolide antibiotics.

The mode of action of the polyene antibiotics is reviewed together with the effect of genetic and environmental factors on sensitive organisms. The future prospects of polyenes in the treatment of systemic mycoses are considered.


The effect of flucytosine on the germination of Candida albicans.

The effect of flucytosine on the germination of 3 strains of Candida albicans in serum was tested. No inhibition of germination was observed and it was concluded that germination did not require the synthesis of DNA.


Problems in the laboratory assessment of antifungal activity.

The end points of tube dilution tests for minimal inhibitory concentrations of miconazole and flucytosine against Candida albicans were difficult to evaluate because partial inhibition was noted over a wide range of antifungal concentrations. This problem was not encountered with amphotericin B. Partial inhibition of Candida arose because of reductions in yeast growth rate and of cell yield. Different sizes of yeast inocula were differentially inhibited by the same concentration of antifungal agent. An in vitro apparatus was described in which miconazole formulated as commercial creams, pessaries and medicated tampons for intravaginal application could be assessed for its inhibitory action in vitro.


Modern aspects of testing azole antifungals.

Difficulties in the in vitro testing of azole antimycotics are described. The value of minimal inhibitory concentration results are discussed and contrasted to those obtained with polyene antimycotics.


Promotion of pseudomycelium formation of Candida albicans in culture: a morphological study of the effects of miconazole and ketoconazole.

The effects of miconazole and its new derivative ketoconazole on Candida albicans have been evaluated by light and electron microscopy. The growth characteristics and morphology of C. albicans in culture for various periods of time in a solution consisting of Eagle's minimum essential medium supplemented with amino acids and fetal calf serum are emphasized. This medium, normally used for culturing mammalian cells, promotes a rather fast growth of C. albicans and favours the development of pseudomycelium. The obvious interest in using such culture conditions for drug evaluation is the prevalence of pseudomycelium, which in vivo is the predominant pathological form of C. albicans. Suppression of pseudomycelium formation is found in the 10-9 to 10-7 M concentration range of the imidazoles. Growth retardation and the destruction of both yeast and pseudomycelial forms brought about by incubating the cells with 10-9 to 10-4 M of the drugs are reported. At low doses these changes include the alteration of cell division, an increase in cell volume and a progressive deterioration of subcellular organelles at the cell periphery. At higher doses the involvement of all other organelles is observed finally leading to complete cell necrosis.


Sensitivity of Candida albicans from patients with chronic oral candidiasis.

Many patients with oral candidiasis respond very slowly or not at all to therapy with amphotericin. Strains of Candida albicans were collected from 17 patients clinically resistant and from 15 who responded to a normal course of amphotericin treatment. Minimal inhibitory concentrations (MIC) determined on diagnostic sensitivity test agar plates gave values of: amphotericin 0-5 mg/l; nystatin 50 i.u./ml; chlorhexidine 12.5 mg/l. No clear MIC could be determined with plates containing miconazole. No difference was noted in MIC values between the 2 groups of patients. Tube-dilution tests in Sabouraud's broth gave MIC values of: amphotericin 0.25 mg/l; nystatin 12.5 i.u./ml; chlorhexidine 1.5 mg/l; miconazole 8-32 mg/l; ketonazole 64 mg/l. Persistence of oral candidiasis is not an indication of infection with resistant organisms. Despite difficulties in in vitro sensitivity testing of miconazole a clinical trial of the drug for treating oral candidiasis is indicated.


Sensitivity of strains of Candida albicans to jaritin, haloprogin, clotrimazole and miconazole.

The antifungal activity of jaritin and haloprogin against Candida albicans is similar. In vitro they had less activity than clotrimazole and miconazole, but the activity was independent of inoculum size.


RNA ligase reaction products in plasmolyzed Escherichia coli cells infected by T4 bacteriophage.

Searching for a physiological role of T4 RNA ligase [polyribonucleotide synthetase (ATP); poly(ribonucleotide):poly(ribonucleotide) ligase (AMP-forming), EC 6.5.1.3] activity, we developed an acellular system of plasmolyzed Escherichia coli cells infected by T4 bacteriophage. Upon incubation of this system with [gamma-32P]ATP, 32P was transferred into a large number of polyribonucleotides, mostly up to 300-400 residues long. The bulk of 32P in the product polyribonucleotides was found in 5'-terminal phosphate groups, suggesting that they originated by a phosphorylation reaction catalyzed by the endogenous polynucleotide kinase (EC 2.7.1.78). Indeed, these products were not seen in an acellular system from uninfected cells, and their amount and complexity increased with the progress of infection. Analysis of the 32P-labeled polyribonucleotide products by gel electrophoresis, either before or after digestion with alkaline phosphatase (EC 3.1.3.1), revealed that a small fraction of the 32P resided in phosphodiester bonds of several tRNA-sized chains. This specific 32P transfer from [gamma-32P]ATP into phosphodiester bonds was apparently catalyzed by successive polynucleotide kinase and RNA ligase reactions. The possible relationship of the 32P transfer to RNA ligase was investigated next by using a system from cells infected with T4 am M69 (an amber mutant deficient in RNA ligase). Transfer of 32P from [gamma-32P]ATP into phosphodiester bonds was not detected in the am M69 system. However, addition of purified RNA ligase to the am M69 system restored the specific 32P transfer. A system from cells infected with T4 psu-b delta 33 (a deletion mutant lacking the entire tRNA region) sustained the specific 32P transfer into tRNA-sized products, indicating that they were not derived from transcripts of T4 tRNA genes. These data may reflect a role of RNA ligase in posttranscriptional conversion of presumably host polyribonucleotides into novel tRNA species during T4 infection.


Initiation of Escherichia coli ribosomal RNA synthesis in vivo.

The 5'-terminal sequences of Escherichia coli ribosomal RNA precursors (pre-rRNAs) synthesized in vivo were characterized by RNA oligonucleotide sequence analysis. The 60- to 170-nucleotide-long 5'-end-specific fragments were produced by RNase III treatment of 30S and 18S pre-rRNAs. Comparison of the RNA oligonucleotides of these fragments with known DNA sequences of the promoter regions of several ribosomal RNA operons allows us to determine the start points of transcription of each operon. We conclude that transcription of most (and perhaps all) rRNA operons is initiated in vivo at two tandem promoters, called P1 and P2, which have recently been identified by in vitro transcription studies of several groups. Depending on the transcription unit, the initiating nucleotide at P1 promoters is either ATP or GTP, whereas at P2 promoters it is either CTP or GTP.


Control of phosphoenolpyruvate-dependent phosphotransferase-mediated sugar transport in Escherichia coli by energization of the cell membrane.

The phosphoenolpyruvate-dependent phosphotransferase-mediated sugar transport in Escherichia coli is inhibited by the energized of the membrane. This was shown in intact cells as well as in membrane vesicles. Relaxation of the proton gradient by uncouplers stimulated the uptake of sugars via the phosphotransferase system in aerobically cultured cells. No such effect was seen in anaerobic cells, apparently because the cell membrane of these cells is poorly energized. Energization by respiration of D-lactate or ascorbate inhibited the phosphotransferase uptake system in membrane vesicles. This inhibition was reversed by the addition of cyanide. Oxamate, a specific inhibitor of lactate dehydrogenase, prevented the inhibitory effect of D-lactate. Membrane vesicles prepared from a cytochrome-less mutant were not energized by D-lactate oxidation and the phosphotransferase uptake system was not inhibited.


Attractants and repellents control demethylation of methylated chemotaxis proteins in Escherichia coli.

A group of methylated proteins, the methyl-accepting chemotaxis proteins (MCP), has been shown to play a central role in bacterial chemotaxis. Both methylation and demethylation of MCP occur continuously in the absence of added stimuli; these two processes are in balance such that a basal level of methylation is maintained. Attractants cause the methylation level to increase to a new value, whereas repellents bring about a decrease in level. Therefore, attractants and repellents must somehow perturb the balance between methylation and demethylation of MCP. In this report the effect of attractants on demethylation of MCP was monitored in two ways: (i) by following the disappearance of [methyl-3H]MCP and (ii) by measuring formation of [3H]methanol, the product of MCP demethylation. Both methods showed that addition of attractants causes a transient inhibition of MCP demethylation. Repellent addition has previously been shown to stimulate MCP demethylation. It is therefore concluded that control of demethylation plays a crucial role in changing the level of methylation of MCP in response to attractants and repellents.


Replication at restrictive temperatures in Escherichia coli containing a polCts mutation.

Escherichia coli cells with a polCts mutation contain a temperature-sensitive DNA polymerase II and fall to replicate DNA at the restrictive temperature (43 degreees C). Mutants deficient in polymerizing activity of the other two recognized DNA polymerases in E. coli can replicate DNA. We have isolated temperature-resistant revertants from a strain containing polA-, polB-, and polCts mutations. These revertants grow at 43 degrees C, but analysis of partially purified DNA polymerase III from several such revertants shows a temperature-sensitive DNA polymerase III activity. Genetic analysis by P1 transduction confirms that such revertants can contain a polCts mutation and also a polA- mutation. We find that such revertants behave phenotypically as PolI+ cells (DNA polymerase I-containing), and extracts of such cells show a DNA polymerase I-like activity. Revertants of polA-, dnaAts and polA-, dnaBts strains do not show such a DNA polymerase activity.


Biotinyl 5'-adenylate: corepressor role in the regulation of the biotin genes of Escherichia coli K-12.

A DNA filter-binding technique was used to study the interaction of the biotin repressor and operator site. From a biotin saturation curve, the concentration for half-maximal binding (K0.5) was calculated to be 1 microM. However, in a similar study with the in vitro coupled transcription-translation system in which biotin served as the corepressor, the K0.5 for repression was 7.1 nM. This marked difference of over 2 orders of magnitude was attributed to the activation of biotin by the partially purified repressor preparation in the in vitro system. The activated product formed from biotin, ATP, and repressor preparation was identified as biotinyl 5'-adenylate by paper chromatography and hydroxamic acid formation. Synthetic biotinyl 5'-adenylate was as effective as biotin in the in vitro system (K0.5, 10 nM) and much more effective than biotin in the DNA-binding assay (K0.5 1.1 nM versus 1 microM). These studies indicate that biotinyl 5'-adenylate has a more direct role in the regulation of the biotin genes than does biotin per se.


Detection of proteins like human gamma and beta globins in Escherichia coli carrying recombinant DNA plasmids.

Escherichia coli strain chi 1776 carrying recombinant DNA plasmids containing cDNA copies of human beta or gamma globin mRNAs has been shown by radioimmunoassay to synthesize polypeptides antigenically related to the beta and gamma chains of human hemoglobin. The gamma and beta polypeptides have been enriched from lysates on immunoabsorbent columns containing hemoglobin antibodies and shown to specifically inhibit the antigen-antibody binding between 125I-labeled hemoglobin and the homologous antibody but not other hemoglobin-antihemoglobin reactions. Clone JW151, which is known to contain a complete copy of the coding portion of the gamma globin mRNA, has been shown to produce a protein that reacts specifically with antibody to the chain of fetal hemoglobin, hemoglobin Kenya, and hemoglobin Bart's.


Isolation and characterization of transducing phage coding for sigma subunit of Escherichia coli RNA polymerase.

A transducing phage has been isolated with codes for the sigma subunit of Escherichia coli RNA polymerase. Transducing phage were selected from E. coli shotgun collections of HindIII or Sac I fragments cloned into Charon 25, a new bacteriophage lambda vector that is capble of forming lyosogens at high temperature. Transduction of an E. coli strain carrying a temperature-sensitive mutation in the sigma gene was used for the selection. The positions of restriction sites for Sac I, HindIII, Xho I, Bgl II, and Kpn I in the cloned bacterial DNA segments were determined. Phage containing the HindIII fragment complement both primase (dnaG) and sigma (rpoD) whereas those containing the Sac I fragment complement only sigma. Results of analyses of the proteins made both in vivo after infection of UV-irradiated cells and in vitro in a coupled transcription/translation system suggest that a Sac I site separates the promoter for sigma from the sigma structural gene. The direction of transcription of sigma was determined to be clockwise with respect to the E. coli genetic map.


Distribution of active and inactive forms of endorphins in rat pituitary and brain.

The recent isolation and identification of alpha-N-acetyl forms of the C-Fragment of lipotropin (beta-endorphin, residues 61-91) and the C'-Fragment (residues 61-87) [Smyth, D.G., Massey, D.E., Zakarian, S. & Finnie, M. (1979) Nature (London) 279, 252-254] has led to a study of their distribution in the pituitary and brain of the rat. Regions were mapped by the method of immunofluorescent staining and the reactive peptides were determined by immunoassay after extraction, gel filtration, and ion exchange chromatography. The major immunoreactive peptides in both lobes of the pituitary were found to be C'-Fragment and N-acetyl C'-Fragment, which are weakly active or inactive as opiates; the C-Fragment and its N-acetyl derivative represented minor components. This indicates that in the rat the circulating "endorphins" released from pituitary would have little morphinomimetic activity. The same four immunoreactive peptides were observed in rat brain. In the hippocampus the C'-Fragment was the principal component in the midbrain there was more C-Fragment but C'-Fragment predominated; in the hypothalamus the C-Fragment was the major peptide, almost to the exclusion of the other peptides. The results demonstrate that the processing of lipotropin is under differential control in anatomically distinct regions of the central nervous system. The processing of lipotropin in the hypothalamus is directed specifically to the production of lipotropin C-Fragment.


Widespread distribution of protein I in the central and peripheral nervous systems.

Protein I, a naturally occurring substrate for cyclic AMP-dependent and calcium-dependent protein kinases, previously has been found only in mammalian brain, where it has been demonstrated to be located in neurons. Various tissues and organs outside the brain have now been examined for the possible occurrence of protein I, by using both an immunohistochemical approach and a chemical procedure involving radioimmunolabeling of polyacrylamide gels. Protein I has been found in the inner plexiform layer of the retina, in the posterior pituitary, and in the autonomic nervous system. In tissue composed predominantly of cells other than nerve cells, immunoreactivity was present only where innervation was present. Protein I appeared to be localized in some, but not all, nerve terminals and synapses.


Immunocytochemical localization, in synapses, of protein I, an endogenous substrate for protein kinases in mammalian brain.

Protein I is a principal endogenous substrate for cyclic AMP- and Ca2+-dependent protein kinases of mammalian brain. Antibodies raised against purified protein I have been used to localize this protein in the rat central nervous system. At the light microscope level, immunoreactivity was detected in punctate deposits in selected zones of synaptic termination. These deposits varied in density among brain regions. At the electron microscope level, immunoreactivity was observed at some but not all synaptic regions and was restricted to the perimeter of synaptic vesicles and to submembranous material in the postsynaptic neuron.


Localization of neurophysin within organelles associated with protein synthesis and packaging in the hypothalamoneurohypophysial system: an immunocytochemical study.

Electron microscopic immunocytochemical localization of neurophysin in the hypothalamo-neurohypophysial system of mice has been studied by using the pre-embedding staining approach to the unlabeled antibody-enzyme technique of Sternberger. In supraoptic cell bodies, peroxidase-antiperoxidase reaction product was localized within cisternae of the nuclear envelope, rough endoplasmic reticulum, and Golgi saccules but not in GERL (Golgi-associated smooth endoplasmic reticulum from which lysosomes arise). Reaction product was also present in secondary lysosomes. Secretory granules in supraoptic perikarya and posterior pituitary Herring bodies were likewise immunoreactive. These findings with the unlabeled antibody-enzyme technique provide conclusive evidence for the localization of an antigen within cellular organelles associated with protein synthesis and packaging.


Alternative secondary structures of leader RNAs and the regulation of the trp, phe, his, thr, and leu operons.

The trp, phe, his, thr, and leu operons of enteric bacteria are regulated by a transcriptional attenuation mechanism. Under conditions of amino acid sufficiency, transcription terminates at an attenuator site after a leader of about 150 nucleotides has been synthesized. Under conditions of limitation of a controlling amino acid, transcription continues past the attenuator into adjacent structural genes. As demonstrated by others, each of the five leader RNAs contains two regions of potential secondary structure which are partially overlapping. One of these regions occurs at the 3' terminus of the leader and is named the "terminator." The other region, which potentially can preclude the formation of the terminator, is named the "preemptor." Conditions that allow the preemptor to form result in derepression. We report here that the five published leader RNA sequences contain an additional potential region of secondary structure, which we call the "protector." The protector partially overlaps the preemptor in such a way that pairing of the former precludes pairing of the latter. For derepression to occur, a ribosome that is translating the leader must block the protector without blocking the preemptor, a condition that is met when the ribosome is arrested at the 3' end of a set of control codons. Including the protector in the model for attenuation explains why derepression of the operon does not result from the arrest of a ribosome at a codon preceding the control set. It also explains why termination is the outcome when transcription occurs in the absence of ribosomes. Finally, termination is the predicted outcome when unfettered translation of the leader RNA occurs, resulting in release of the ribosome at the translational stop signal.


Effect of diethylpyrocarbonate on lactose/proton symport in Escherichia coli membrane vesicles.

Exposure of Escherichia coli ML 308-225 membrane vesicles to the histidine-specific reagent diethylpyrocarbonate (DEPC) led to concentration- and time-dependent inactivation of active lactose transport, and the sensitivity of the system to inactivation was enhanced when an electrochemical proton gradient (delta- muH+, interior negative and alkaline) was generated across the vesicle membrane. Although beta-D-galactopyranosyl 1-thio-beta-D-galactopyranoside blocked DEPC inactivation, binding of p-nitrophenyl alpha-D-galactopyranoside was not significantly altered, indicating that DEPC does not react at the binding sites of the lac carrier protein. Strikingly, vesicles treated with DEPC exhibited an increased apparent Km for delta- muH+-driven lactose transport and counterflow but no change in the Vmax of these reactions and no change in the apparent Km or Vmax of facilitated diffusion. Moreover, DEPC treatment increased the apparent Km observed for delta- muH+-driven proline and D-lactate transport with no change in Vmax. Finally, the lactose counterflow activity of DEPC-treated vesicles was regenerated by subsequent exposure to hydroxylamine. It is suggested that a histidyl residue(s) in the lac carrier or another protein in the translocation complex is involved either in the binding and translocation of protons or in a conformational change that may occur upon protonation of the lac carrier protein.


Cloning and expression of the Escherichia coli replication origin in a single-stranded DNA phage.

The Escherichia coli DNA replication origin (oriC) and the adjacent asparagine synthetase gene (asnA) have been inserted into the duplex replicative form DNA of the single-stranded phage vector M13Goril. By in vitro recombination, the entire oriC asnA-containing plasmid pJS5 was inserted into M13Gori1 in both possible orientations. Both phage types transduce the asnA gene and confer upon the M13 vector the ability to replicate as a plasmid in the E. coli mutant rep3. In rep+ hosts, these phages undergo single-stranded DNA synthesis and viral morphogenesis.


Attractants and repellents influence methylation and demethylation of methyl-accepting chemotaxis proteins in an extract of Escherichia coli.

During bacterial chemotaxis, attractants and repellents alter the methylation levels of the methyl-accepting chemotaxis proteins (MCPs). These methylation levels represent a balance between two enzymatic processes: methylation and demethylation. In vivo experiments previously have shown that chemoeffectors influence the demethylation process; effects on the methylation system have not been reported. Here we show that in a cell-free extract of Escherichia coli both methylation and demethylation of the MCPs are affected by attractants and repellents. Attractants enhance methylation and inhibit demethylation. Repellents inhibit methylation and stimulate demethylation. The cell-free system provides an opportunity for further study of the mechanisms by which attractants and repellents influence the levels of methylation of the MCPs.


Molecular cloning of Moloney murine sarcoma virus: arrangement of virus-related sequences within the normal mouse genome.

The unintegrated circular DNA form of Moloney murine sarcoma virus (MSV) has been cloned in bacteriophage lambda. Discrete deletions in the viral genome were shown to occur during propagation of recombinant phage in Escherichia coli. Heteroduplex and restriction enzyme analyses indicated the deletion of tandemly repeated sequences within certain of the cloned MSV DNA inserts. Cloned MSV DNA was used to prepare a probe composed of its acquired cellular (src) sequences, shown previously to be necessary for MSV transformation. Analysis of EcoRI digests of normal mouse cellular DNA revealed the presence of a single 14-kilobase-pair fragment containing these sequences which lacked contiguity with endogenous type C helper viral information of the same cells. Thus, the sarcoma virus-specific sequences of MSV are represented within the normal mouse genome in a manner analogous to that of a cellular gene.


Organization of spacer DNA in chromatin.

Detailed analysis of the DNA fragment patterns produced by DNase I digestion of yeast, HeLa, and chicken erythrocyte nuclei reveals surprising features of nucleosome phasing. First, the spacer regions in phased yeast chromatin must be of lengths (10m + 5) base pairs, where m = 0, 1, 2,.... This feature is not seen in parallel studies of chicken erythrocyte chromatin. The 5-base pair increment in the yeast spacer imposes interesting restraints on the higher order structure of yeast chromatin. Second, we have been able to simulate the DNase I cutting patterns and get good agreement with the observed yeast patterns. Third, three different chromatins show a long range periodicity in the DNase I digest pattern, with a period half that of the staphylococcal nuclease repeat. These results suggest that the amount of chromatin observed in discrete extended-ladder bands is a minimum estimate of phasing and in fact phasing may be a more general feature.


Nucleosome organization of the yeast 2-micrometer DNA plasmid: a eukaryotic minichromosome.

The eukaryotic microorganism Saccharomyces cerevisiae contains 50-100 copies per cell of a circular plasmid called 2-micrometer DNA. The intracellular structure of these molecules, which represent about 4% of the total DNA, was examined by digestion of total cellular chromatin with micrococcal nuclease (nucleate 3'-oligonucleotidohydrolase, EC 3.1.31.1). Nuclease-resistant DNA fragments were fractionated by gel electrophoresis and 2-micrometer DNA sequences were detected by hybridization. The 2-micrometer and chromosomal DNA digestion patterns were very similar indicating that both types of DNA are condensed into nucleosomes. An analysis of these digestion patterns showed that the kinetics of digestion of 2-micrometer chromatin and total chromatin are similar and that both have the same nucleosome repeat length of about 165 base pairs. Native 2-micrometer plasmids were examined by zone sedimentation in sucrose gradients containing 0.15 M NaCl and were found to have a sedimentation constant of 75 S, about 3 times the sedimentation constant of protein-free 2-micrometer DNA. This sedimentation property is what would be expected for a 2-micrometer DNA minichromosome. We conclude that within the cell 2-micrometer DNA molecules are organized in a chromatin structure very similar to that of the yeast chromosomes.


Genetic control of enhanced mutability of mitochondrial DNA and gamma-ray sensitivity in Saccharomyces cerevisiae.

Five nuclear mutants enhancing the spontaneous mutation rate of mtDNA have been isolated in Saccharomyces cerevisiae. These mutators fall into five complementation groups and are located at five genetic loci different from rad50 to rad57 loci. Three mutants (gam1, gam2, and gam4), insensitive or weakly sensitive to gamma-rays, exhibit increased frequency of spontaneous production of mutants with large deletions of the mtDNA (p-) and of all tested mitochondrial drug-resistant mutants. Two other mutants (gam3 and gam5), highly sensitive to gamma-rays, increase only the mutation rate of particular alleles of the mtDNA. The mutant gam5 enhances only the production of p- and erythromycin-resistant clones. The mutant gam3 exhibits an enhanced rate of oligomycin-resistant clones as well as a collateral increase of nuclear mutability. The existence of gam3 and gam5 mutants indicates that at least two common steps control both nuclear DNA repair and the mutability of particular alleles of the mtDNA. However, the general spontaneous mutability of the mtDNA includes at least three steps not involved in the repair of nuclear DNA, as revealed by the gam1, gam2, and gam4 mutations.


Analysis of HLA-D region-associated molecules with monoclonal antibody.

A group of monoclonal antibodies directed against nonpolymorphic HLA-D antigens (human Ia antigens) was produced by somatic cell hybridization. One of these antibodies was used to analyze the two-dimensional polyacrylamide gel electrophoretic pattern of the HLA-D alloantigens from different HLA-D homozygous B-cell lines. Two-dimensional gels of [35S]methionine-labeled cell extracts immunoprecipitated by a monoclonal antibody directed against HLA-D antigens showed a complex set of spots whose electrophoretic pattern varied according to the HLA-D type. The major HLA-D-related electrophoretic polymorphism was found in the basic and smaller (26,000-28,000 daltons) HLA-D polypeptides. These patterns represent allele-specific "fingerprints" of different HLA-D genotypes. There are striking similarities with respect to number, size, and charge spectrum among HLA-D polypeptides and the murine I-A and I-E subregion polypeptides, suggesting a similar genetic organization and molecular complexity in both species.


Rich ependymal investment of luliberin (LHRH) fibers revealed immunocytochemically in an image like that from Golgi stain.

Immunospecific staining in 100-micron-thick Vibratome section by the unlabeled antibody method resembles Golgi staining and reveals an abundance of luliberin- (luteinizing hormone releasing hormone, LHRH) positive cells and fibers in close contact with the surface of the third ventricle. The polarity of LHRH cells can be seen and processes can be traced for several millimeters. In the medial preoptic and suprachiasmatic areas bipolar LHRH neurons send short stout processes to the ventricular surface and long processes toward the organum vasculosum laminae terminalis. These cells resemble the receptor cells contacting the cerebrospinal fluid that have been described by Vigh and Vigh-Teichmann [Vigh, B. & Vigh-Teichmann, I. (1973) Int. Rev. Cytol. 35, 189-251]. In the septal region some bipolar neurons send both of their processes towards the ventricular surface. LHRH neurons in the nucleus of the anterior commissure and the bed nucleus of the stria terminalis project over the anterior commissure to form a dense plexus of fibers in the subfornical organ. The proximity of LHRH perikarya and fibers to the ventricular surface supports the hypothesis [Knigge, K.M., Joseph, S.A., Scott, D.E. & Jacobs, J.J. (1971) in The Neuroendocrinology of Human Reproduction, eds. Mack, H.C. & Sherman, A.J., (Thomas, Springfield, IL), pp. 6-22.] that the cerebrospinal fluid functions in neuroendocrine control mechanisms.


Salt, hypertension, and genetic-environmental interactions.

Observations relating to hypertension in humans and rat experiments have been reviewed from several viewpoints including clinical medicine, pathophysiology, epidemiology, and genetics. It seems likely that much of essential hypertension results from excessive salt intake by individuals with an inherited inability to excrete sodium efficiently which is compensated by blood pressure elevation. A few major genes are likely responsible for a few basic mechanisms involving renal membranes and hormones controlling sodium transport. Excess hypertension in blacks, diabetics, the elderly, and oral contraceptive users can be explained by this theory. If hypertension and other genetically predisposed CHD risk factors are to be fully understood, future studies are needed with detailed data on both genetic and environmental factors, and analytic tools that allow an adequate examination of their interactions.


An easily constructed durable chronic intracerebral cannula system.

A simple, inexpensive, and rugged intracerebral cannula system is described and construction details are provided for both guide and injection cannula assemblies. The guide cannula assembly is easily and quickly molded from dental acrylic cement and has a protective acrylic collar surrounding the end of the actual guide cannula. This system is believed to offer some advantage over current methodologies in terms of expense, durability and ease of construction and is well suited to use in rodents.


Functional components of surface membranes: potential targets for pharmacological manipulation.

This short survey of three major developments in membrane studies should serve to convince pharmacologists that biochemical approaches to the study of cell membranes have finally borne fruit. A reasonable model of cell membrane architecture is now in hand, and it should be possible to use these findings to design and interpret new pharmacological approaches to the manipulation of cells via their surface membranes.


Accumulation of acetylcholine receptors and acetylcholinesterase at newly formed nerve-muscle synapses.

Nerve-muscle synapses form and the postsynaptic membrane begins to differentiate rapidly in vitro. New clusters of AChRs have been located at transmitting synapses within 3 hours after a competent neurite contacted a receptive myotube. Receptors within new subneural clusters are not identical with receptors at adult end-plates: they are metabolized rapidly and their mean channel open time is prolonged. In this sense synapse formation must be considered a drawn out affair. AChE also accumulates at synapses soon after functional contacts are established. The cleft enzyme is apparently regulated differently than receptors in the postsynaptic membrane. Muscle activity is necessary for the early accumulation of AChE, but not for the clustering of receptors. Further studies in vitro should be useful for analysis of the role of muscle activity, of the local neural influence on the postsynaptic membrane, and of the mechanism of the muscle response.


Hepatic clearance measurements and pharmacokinetics.

This review emphasizes the need for well-defined models for hepatic pharmacokinetics and discusses the interpretation of hepatic clearance measurements. Such recent approaches are subjected to theoretical and experimental comparisons. The pharmacokinetic consequences, including a classification of drugs according to hepatic clearance, are outlined.


Genital herpes simplex virus infections.

In recent years, a great increase in interest in genital herpes has been stimulated partly by the rising prevalence of this disease and partly by observations suggesting that genital herpes is a cause of cervical cancer. The clinical pictures produced by genital herpes simplex virus infections are similar in men and women. In contrast to recurrent attacks, initial episodes of infection are generally more extensive, last longer, and are more often associated with regional lymphadenopathy and systemic symptoms. Genital herpes in pregnancy may pose a serious threat to the newborn infant. Although the data suggesting genital herpes simplex virus infection is a cause of cervical cancer are quite extensive, the evidence is largely circumstantial. In spite of these more serious aspects of genital herpes simplex virus infection, episodes of genital herpes are almost always self-limited and benign. Frequent recurrences pose the major therapeutic and management problem. At present, there is no satisfactory treatment for recurrent genital herpes simplex virus in fection. Many of the suggested therapies, although some sound very promising, are potentially dangerous and should be used only under carefully controlled conditions.


Nephrotic syndrome.

The nephrotic syndrome appears to be an expression of a number of complex disease processes. The supportive, as well as the definitive, management of the nephrotic syndrome patient requires considerable knowledge of disease pathophysiology, drugs, and drug interactions. Therapy of any degree cannot be undertaken lightly for fear of further compromising the nephrotic syndrome patient.


Stinging insect allergy.

One to two million Americans have a history of systemic allergic reactions to stinging insects and must live with the knowledge that future stings could be catastrophic. Recent advances have been made in diagnosis and treatment of this problem. Pure venoms have been recognized to be superior to the whole body extract as diagnostic and treatment antigens. Use of pure venoms for immunotherapy gives far better protection than whole body extract for the allergic patient and represents a major advance in the field of allergy. The imported fire ant is very different from the other common stinging insects in its venom components, the type of local reactions caused by its sting, and the fact that whole body extract from this insect seems to be as good an antigen for diagnosis and treatment as pure venom. Although important discoveries about insect allergy have been made recently, some very important questions are yet to be answered. Among these are: (1) Can suppliers of venom provide us with enough venom to treat all patients with insect allergy? (2) How long should immunotherapy be continued? and (3) Would treatment with specific antigens from venom be superior to using whole venom? We all await the answer to these and other questions with hopeful anticipation.


Social forces, ideology, and the domain of mental health services.

Public attitudes toward the mentally ill affect practices relating to treatment of mentally impaired persons. This has been true historically. The human rights movement and the community mental health movement have had a major impact on the care and treatment practices of the twentieth century. In the United States these movements have influenced Federal policy, which in turn has provided resources and a redirection that heralded a major expansion of public mental health services.


Benjamin Rush and his friends on human nature.

The 18th century writers explored the nature of man and decided mankind was divided into six subdivisions. Studying man from a naturalistic viewpoint raised doubts about God's primary role, a questioning totally unacceptable to Benjamin Rush. Rush considered all of mankind as originating from God's original pair, but bearing a deficiency and vulnerability from their original sin. He viewed man as a physiologist, as a materialist, and as a monist. Modifying the Scottish philosophers extensive faculty psychology, he accepted nine mental faculties and of these considered the moral sense essential. Rush and his friend, Thomas Jefferson, shared this belief which enabled them to be more optimistic than their friend John Adams who thought man to be driven by a need for distinction and thereby requiring control from a strong central government.


Prospective study of different methods and routes of administration of prostaglandin E2 to improve the unripe cervix.

Methods of vaginal and extra-amniotic prostaglandin administration to achieve ripening of the cervix as a preliminary to induction of labour are described. Three groups of twenty patients with unfavourable induction features were studied, each receiving prostaglandin E2 the evening prior to planned induction. One group received PGE2 500 micrograms suspended in a viscous medium extra-amniotically. One group received PGE2 3 mg suspended in a viscous medium into the vaginal vault. A third group received a 3 mg PGE2 vaginal pessary to the posterior fornix. Improvement in cervical status at time of induction occurred in all groups but no single group had a significant advantage when regarding mean improvement, the induction-delivery interval or the number of patients in whom labour began before formal induction. However, with regard to relative cost, ease of preparation and storage, as well as patient and medical staff convenience, Prostaglandin E2 in pessary form is a superior form of administration.


Effect of ibuprofen on menstrual blood prostaglandin levels in dysmenorrheic women.

In a randomized crossover study 15 dysmenorrheic women were treated during two consecutive menstrual period, once with the potent prostaglandin-synthesis inhibitor: ibuprofen and once with an identical looking placebo. Each patient was medicated for 12 hours during the first day of her menstrual flow and was subsequently fitted with a cervical cup for the collection of menstrual blood during three hours. In these samples the concentrations of prostaglandin (PG)F and PGE were measured by radioimmunoassay. The patients receiving placebo had high PGF levels 135 +/- 27 ng/ml (Mean +/- S.E.) which were significnatly reduced by Ibuprofen to 24 +/- 5 ng/ml (P less than 0.001). The PGE concentrations decreased from 5 +/- 1 ng/ml to 2 +/- 1 ng/ml (P less than 0.05). Ibuprofen also reduced the menstrual pain significantly (P less than 0.001). These results substantiate the earlier conclusion that a causal relationship exists between effective treatment with PG-synthesis inhibitors and decrease in menstrual blood PG levels, intrauterine pressure and dysmenorrheic pain.


The effect of prostaglandin F2 alpha on third stage labor.

The effects of intramyometrially injected PGF2 alpha intravenous Ergometrin and no treatment were compared during 3rd stage labor in 140 patients. In comparison with Ergometrin or no treatment, PGF2 alpha significantly reduced the duration of 3rd stage labor, blood loss, incidence of subinvolution and subfebrility.


Controlled trial of induction of labor by vaginal suppositories containing prostaglandin E2.

A group of 84 women at 39-43 weeks of pregnancy were randomly allocated to a blind trial of induction of labor with vaginal suppositories containing inert material or either 0.2 mg or 0.4 mg of prostaglandin E2. The suppositories were self-administered every two hours during waking hours on two successive days until labor started or 15 had been used. Side-effects were absent. Labor was established within 48 hr of insertion of the first suppository in 9.3% of control patients, 65.4% of those treated with 0.2 mg PGE2 and 85.7% of those treated with 0.4 mg PGE2. The mean Apgar scores in the three groups were the same. The mean total dose of PGE2 were 2.0 mg (0.2 mg group) and 2.3 mg (0.4 mg group). It is concluded that vaginal PGE2 is an effective and acceptable method of inducing labor at term.


The central nervous effect of prostaglandins I2, E2 and F2 alpha on aconitine-induced cardiac arrhythmia in rats.

Intracerebroventricular administration of PGI2 or PGE2 reduced aconitine-induced cardiac arrhythmia in rats. PGF2 alpha had no antiarrhythmic effect under the same conditions. The ED50 values of PGI2 and E2 were 0.25 microgram/kg and 1.1 microgram/kg, respectively. Central mechanisms may participate in the antiarrhythmic effect of these PGs.


[Cartesianism and Henricus Regius' dentistry (1598--1679--1979) (Henricus de Roy, René Descartes].

Henricues Regius (1598--1679), professor at the University of Utrecht was the first physician who accepted the physiology of the philosopher Descartes (1596--1750) that he exposed in Fundamenta physices (1646) and in Fundamenta Medicinae (1647) but in Praxis Medica (1657) his therapy of the "odontalgia" is still mediaeval according to the principles of Galen.


[Odontology and the beginning of cartesianism (1673--1650) (Rene Descartes)].

In the seventeenth century the universities of the Netherlands underwent the influence of Descartes in all the faculties. In medicine three periods can be distinguished: in the first, pathology and therapy are still galenic; the second, by the application of the cartesian method, triumphs in physiology; and the third, corrected by the views of Newton is integrated in a moderate biomechanism.


[Evoked potentials and vigilance states induced during the course of choice reaction time tests].

Characteristics of Visual Evoked Potentials (N120, P200, P300) were investigated during choice reaction time situations in a group of 10 subjects practising meditation (E.S.) versus a matched control group (C.S.) During a series of visual stimuli occuring at fixed intervals, with 10% random omissions, the subjects were asked : 1) to respond by a finger displacement to each visual stimulus; 2) to hold on the response to the stimulus and to respond to omission. Both tasks were recorded before and after the practice of meditation or rest for the controls. The intergroup comparison showed that the experimental subjects had faster RT's with less mistakes, and N120 and P200 of larger amplitude and shorter latency. These differencies were significant before and after meditation. The transient effects of meditation or rest, were opposite for the two groups : whereas after meditation the RT's became longer with less mistakes, and the amplitude of P300 larger, after rest there was a decrease of the P300 amplitude and no change in the RT's of the controls. These results are interpreted in terms of selective attention capacity and information processing strategies, A.S.C. being used as a model for the study of these processes.


Bismuth toxicity in man II. Review of bismuth blood and urine levels in patients after administration of therapeutic bismuth formulations in relation to the problem of bismuth toxicity in man.

A survey of the leterature on bismuth toxicity in man in relation to blood level data, has revealed the necessity of distinguishing between lipid soluble and water soluble organic complexes of bismuth on the one hand and the simple inorganic salts of bismuth on the other hand. A characteristic feature of the former, illustrated by the water soluble bismuth complex triglycollamate, is the high bismuth levels (due to absorption of the complex as such) and the nephrotoxic properties of the compound in man. Bismuth absorption after administration of the simple inorganic salts of bismuth is postulated to occur in the form of ionic bismuth as such, low bismuth levels being characteristic features of such compounds. Bismuth blood and urine levels obtained from patients after administration of a new anti-ulcer drug (Bicitropeptide) in a well controlled clinical trial are discussed and suggest that that this bismuth containing drug behaves pharmacologically in a manner similar to the inorganic bismuth salts in man, low bismuth blood levels and the absence of toxic side effects being conspicuous features of the drug. Based on these considerations, it is proposed that the pharmacologically active bismuth compounds be divided into four different groups depending on structure, stability and solubility. The question as to what constitutes a "toxic bismuth blood level" can only be discussed in relation to the new proposed sub-division of bismuth compounds and is only meaningful if the term is defined to relate only to ionic bismuth (presumably bound to a large extent to blood proteins). Based on information gleaned from the literature and blood level values reported in the clinical trial referred to, it is suggested that bismuth blood level values below 50 micrograms/ml are highly unlikely to be associated with meaningful toxicity in man. Finally, attention is drawn to the reversibility of bismuth toxicity in man as reported by many authors irrespective of the type of bismuth compound concerned.


The effect of dietary disulfiram upon the tissue distribution and excretion of 14C-1,2-dibromoethane in the rat.

Dietary disulfiram enhances the toxicity of inhaled 1,2-dibromoethane in rats. This study was undertaken to determine whether the differential toxicity noted was associated with alterations in the levels of the compound and/or its metabolites in the target organs. A comparison of the levels of 14C in selected tissues of male rats, with and without dietary disulfiram, following the oral administration of 14C-1,2-dibromoethane was made. The results indicated that levels of radioactivity in the target organs of animals in  the disulfiram group were significantly elevated both at 24 and 48 hours following compound administration. The data indicate a direct correlation between tissue levels and the enhancement of toxicity noted in the disulfiram-treated rats in the inhalation study. A significant elevation in the levels of radioactivity in washed liver nuclei obtained from animals receiving dietary disulfiram was also noted, suggesting a relationship between nuclear uptake and the increased incidence of liver tumors appearing in the disulfiran group in the inhalation study.


Effects of some naturally occurring prostaglandins of the D-, E-, and I-type and synthetic analogues on adenylate cyclase of human fat cell ghosts.

The effects of various prostaglandins originating from arachidonic acid (prostaglandin E2, D2, I2) as well as of methylated analogues of prostaglandin E2 (16,16-dimethyl prostaglandin E2 and 15-S-methyl prostaglandin E2) on the human fat cell adenylate cyclase were studied. In addition, the stable breakdown product of prostaglandin I2, prostaglandin 6-keto-F1-alpha, was tested for activity. Prostaglandin E2 and its methylated analogues as well as prostaglandin D2 and I2 were active stimulators of the human fat cell enzyme. The stable breakdown product of prostaglandin I2 (prostaglandin 6-keto-F1-alpha) which is biologically inactive had no stimulatory effects, suggesting that stimulation of the human fat cell function. The stimulatory action of prostaglandin D2 is consistent with the elevation of cAMP-levels induced by this compound in intact fat cells. It is concluded that the stimulatory effects of prostaglandins on the human fat cell adenylate cyclase are related to their stimulatory effects upon lipolysis.


[Canine kidney preservation by intermittent perfusion with hypothermic Collins- or Sacks-solution (author's transl)].

37 canine kidneys were intermittently perfused with Collins- or Sacks-solution every 2--6 h. 32 of these kidneys were transplanted after 12--24 h storage time. Additionally, 24 kidneys were stored under hypothermia for 12--24 h and then transplanted (control group). During perfusion the oxygen consumption of the kidneys and the enzyme and lactate release were measured. It was impossible to improve the results of hypothermic storage preservation by intermittent perfusion of the kidneys, on the contrary kidney function deteriorated by the number of perfusion processes. The failure of intermittent perfusion is caused by the increase of renal vascular resistance during perfusion. From the oxygen consumption and lactate release measurement it was concluded, that the washout solutions for kidney preservation should contain more substrate which can be utilized under anaerobic conditions to improve the results of hypothermic storage preservation.


Ultrastructure of skeletal muscle after CO2-laser incision.

The immediate effect of CO2-laser incision to the dorsal skin and underlying muscular tissue of rats was studied by light and electron microscopical methods. In the subcutaneous layer of skeletal muscle cells an increased susceptibility was found, resulting in distinct zones of cellular changes which are attributed mainly to thermal effects of the laser beam. The importance of these findings for the surgical application of lasers is discussed.


[Adenosine deaminase activity in blood cells; influence of cytostatic therapy in patients with acute leukemia and with renal transplants (author's transl)].

A causal relation between the enzyme, adenosine deaminase (ADA), and immune dysfunction is well known: patients with congenital inactivity of ADA invariably suffer of severe combined immunodeficiency. In contrast, we found in patients treated with immunosuppressive drugs increased ADA enzyme activity. Previous findings on ADA activity in acute leukemias are until now controversial. We found normal to increased ADA activity in children with acute lymphatic leukemia (ALL) and acute myeloid leukemia (AML) in remission as long as they were treated with cytostatic drugs. In the group of cured leukemics (in continuous remission after suspension of the therapeutic regimen) the ADA activities were normal. These findings do not exclude a heterogeneity within the leukemia group. They do not explain the signs of cellular immunodeficiencies well known in patients with acute leukemias.


Preliminary studies on cholinergic control of pituitary gonadotrophin secretion in the ewe.

Administration of neostigmine, a cholinomimetic, to anoestrous ewes diminished the increase in jugular plasma luteinising hormone and follicle-stimulating hormone induced by oestradiol. Intravenous injection of atropine had no effect on the gonadotrophin surge.


Increase in resistance to acute experimental coli-septicaemia in chicks given high levels of ferrous sulphate in the diet.

The resistance of chicks to coli-septicaemia following injection of approximately 10(8) cells of Escherichia coli O78 was found to be appreciably increased in terms of mortality by the supplementation of their diet with 360 mg/kg or more of iron in the form of commercial hydrated ferrous sulphate. This was related to an increase in the plasma iron levels. The results suggest that increased resistance to infection could be partly achieved by correcting the hypoferraemia which has previously been shown to be produced in chickens by injections of E coli endotoxin lipopolysaccharide (LPS). The results also showed that haematological disturbances in survivors of coli-septicaemia can persist for up to at least 30 days after infection.


Radioimmunoassays of human myoglobin in serum and urine.

Two solid-phase radioimmunoassays have been developed for the detection of myoglobin in serum and urine. The sensitivity of the methods is 0.1 and 0.5 microgram/l, respectively, with a coefficient of variation of the respective method of 7-8%. The mean serum concentration of myoglobin in ninety-nine healthy blood donors was 44.3 microgram/l +/- 18.0 microgram/l (SD) with a significant difference (P less than 0.001) between men (50.6 +/- 19.8) and women (35.7 +/- 10.4). Serum myoglobin was positively correlated to age (P less than 0.05), body weight (P less than 0.02), serum creatine kinase (P less than 0.001), and serum creatinine (P less than 0.001) to galactose elimination rate. Serum myoglobin levels were not influenced by exhaustive short time dynamic exercise. The mean urinary excretion of myoglobin in twenty-four healthy students was 1.2 microgram/24 h (range 0.1-4 microgram/24 h). Myoglobin excretion was correlated to excretion of beta 2-microglobulin (P less than 0.02) but not to serum levels of myoglobin. No indications of circulating antibodies to myoglobin were obtained when assaying sixty-seven rheumatoid arthritis and thirteen myastenia gravis sera. Presence of other myoglobin binding substances in serum, which would interfere with the assays also seemed unlikely. Determination of myoglobin in serum by sensitive and specific method might be of clinical value in the diagnosis of diseases involving muscle tissues.


Simple immunofixation modification: a practical way to identify monoclonal protein fractions.

A simple immunofixation modification for identification of monoclonal protein fractions has been adapted on gelatinized cellulose acetate membranes. This modification offers excellent sensitivity, simplification of interpretation and low costs. The analysis result is available in less than 3 h.


Comparison of different measurements of development of plaque and gingivitis in man.

The purpose of the present study was to compare efficiency of measurements commonly used to study the development of plaque and gingivitis in short-term clinical trials. Data from a recent experiment on the effect of mutanase on accumulation of plaque and development of gingivitis were used. Efficiency was compared using 1) level of significance and 2) minimum number of individuals necessary to obtain a statistically significant result at a given level of significance and with a given power of the test. Of all the measurements used, the gingival exudate measurement was the most efficient. Among the nonparametric indices, Plaque Index was the most efficient, especially on proximal surfaces in the mandible. Reduction of the scales for the nonparametric indices from a four-point scale to a two-point (dichotomous) scale slightly reduced experimental efficiency.


Arrangement of actin filaments in in vitro migrating human gingival epithelial cells.

Using immunofluorescent techniques and anti-actin antibodies the presence of actin filaments has been demonstrated in epithelial cells cultivated in vitro. Organized bundles of filaments are restricted to the marginal cells in epithelial sheets. These marginal cells are not contact-inhibited and show structures like membrane ruffles which are common for migrating cells. In contrast, internal cells in the epithelial sheets are diffusely stained by anti-actin antibodies. The actin in all cells is redistributed after incubation in media containing cytochalasin B (CB). The importance of organized actin-filaments in migrating epithelial cells is discussed.


Effect of caries in mentally handicapped children of addition of fluoride and bicarbonate-phosphate to dietary sugar products.

Mentally handicapped children, aged 5--15 years and living in institutions, received fluoride supplement in several sugar products of their diet; in candies, marmalades, jams, fruit juices and in sweet desserts corresponding to 10 mg F as NaF per kg of the sugar (sucrose or glucose) of each product. To two of the four daily candies was also added a NaHCO3 + KH2PO4 mixture (mole ratio 9.8/l, resp.) to substitute for 2.5% of the sugar of the candy. The control children received the respective products without the additives. After stepwise exclusions of subjects for various reasons, e.g. for the absence of permanent teeth, low initial caries activity, strong medication, Down's syndrome, etc., the mean DMFS-increment in the remaining 43 control subjects was 4.5 and in the 41 test subjects 2.6 lesions/100 surfaces at risk, i.e. 42% reduction. Caries arrestment had occurred in these test subjects after the first year, while in the respective controls it was continuously increasing. Among numerous oral and body parameters studied, only surface enamel fluoride in primary teeth was increased by the fluoride supplements and urinary phosphate and calcium excretion decreased.


Age differences in cortical width of mandibles determined by histoquantitation.

A study was carried out to develop a histomorphometric method for indication of individual level of cortical width (MCW) and absolute cortical bone mass (MCW X % bone mass in mm3/mm2 bone surface) in the mandibular body and to demonstrate possible differences as to site, side, state of dentition, sex and age. The material consisted of autopsy specimens of mandibles. Microradiograms of 100-microns-thick buccolingual ground sections were used. An electronic point-counting system (Leitz-Classimat) was used for determination of MCW. The values for percentage of cortical bone mass have been given in previous studies. The analysis indicates that: 1) a reproducible measure for MCW in micron and absolute bone mass in a standard site can be obtained; 2) MCW in other sites of mandibular cortex deviates from that of standard site, which is why the same site has to be used in interindividual analysis; 3) MCW and absolute bone mass in the standard site are independent of side, state of dentition and sex, but show age-related decreases, while 4) the lingual cortex is unsuitable for interindividual analysis of MCW.


An electron-microscopic study of human thyroid cancer.

Authors studied the ultrastructural characteristics of the following thyroid cancer: papillary carcinoma, follicular carcinoma, undifferentiated carcinoma and medullary carcinoma. Some specific ultrastructural-functional correlations for each type of thyroid cancer could be established. Papillary and follicular carcinoma had some common features: larger nuclei than in benign lesions, a highly increased number of mitochondria, a reduced endoplasmic reticulum, cell junctions between the cells and an intact basal lamina. In addition, papillary carcinoma presented stage I and stage II nuclear inclusions, and nuclear invaginations that contained cytoplasm. The higher malignancy of follicular carcinoma compared with that of papillary carcinoma was assigned to less differentiated areas corresponding to the compact fields. Undifferentiated carcinoma consisted of large pleomorphic cells (spindle and giant cells) with abundant mitochondria, a flat rough endoplasmic reticulum, scanty secretory granules and lysosomes, cell junctions, all suggesting their common epithelial origin. Ultrastructure of medullary carcinoma contributed to the explanation of the amyloid origin and of granule types in correlation with hormone storage in cells.


Scanning electron microscopy at macromolecular resolution in low energy mode on biological specimens coated with ultra thin metal films.

In this report, conditions for attaining high resolution in scanning electron microscopy with soft biological specimens are described using the currently available high resolution scanning electron microscopes in emission mode of low energy electrons (secondary and charging electrons). Retinal rod outer segments, red blood cells, intestinal mucosa, and ferritin molecules were all used as biological test specimens. From uncoated specimens a new source of signal, referred to as a discharge signal, can provide a high yield of low energy electrons from an excitation area approximately the size of the beam's cross section. Additionally, under these conditions sufficient topographic contrast can be achieved by applying ultra thin metal coatins. A 0.5 nm thick gold film is found sufficient for generating the total signal, whereas increased coating thickness causes additional topographic background signal. However, a 2.0 nm film is needed for imaging surface details with the present instrument. Ultra thin, even, and grainless tantalum films have been found effective in eliminating the charging artifacts caused by external fields, and the decoration artifacts caused by crystal growth as seen in gold films. To improve, in high magnification work on ultra thin coated specimen, signal-to-noise ratio, methods for obtaining saturation of the signal with discharge electrons are shown. The necessity of confirming the information obtained in SEM by independent techniques (TEM of stereo-replicas or ultra thin sections) is discussed.


Correlation by scanning electron microscopy of in vitro and in vivo effects of amoxicillin and ampicillin on the morphology of Escherichia coli.

The effect of in vitro incubation with subinhibitory and inhibitory doses of amoxicillin or ampicillin on the morphology of E. coli 257 was evaluated by scanning electron microscopy (SEM). No effect, with either penicillin was seen at 1/16 X MIC (0.39 microgram/ml). Filament formation was the major effect seen with both agents at 1/8, 1/4 and 1/2 X MIC but only with ampicillin at the MIC (6.25 microgram/ml). Cells incubated with amoxicillin at the MIC were primarily short rods with bulbous swellings. This morphological difference between the two antibiotics was dose-related since ampicillin, at 2 X MIC, also caused the formation of short rods with bulbous swellings. SEM examination of omentum from amoxicillin and ampicillin treated mice 2 h after infection with E. coli 257 indicated a correlation of morphological effects seen in vitro and in vivo with the respective antibiotics. For example, following a single subcutaneous treatment with amoxicillin and ampicillin at 250 mg/kg, the plasma concentrations 2 h later (the time of infection) were 8.2 and 7.7 microgram/ml respectively but short rods with bulbous swellings were seen in amoxicillin-treated mice whereas filaments were found in the ampicillin-treated mice.


Preparation and observation by SEM of hemopoietic cells cloned in soft agar.

A method is described to prepare clones of hemopoietic cells grown in soft agar for scanning electron microscopy (SEM). A critical modification of the otherwise quite standard SEM processing procedure for biological samples involved the use of silver micropore disks as an adherent substrate to support the highly labile, deformable agar slabs. This support allows maintenance of the normal flat pancake shape of the specimen through the thiocarbohydrazide osmium ligand binding steps, dehydration, and critical point drying. With this support and careful dissection of the surface agar with a fine steel needle using a stereomicroscope, selected areas and depths within the colony can be exposed and examined by SEM. Surface topography of cloned cells can be correlated with intracellular cytological features by excising areas of interest and directly embedding them in plastic for thin-section preparation and viewing by transmission electron microscopy (TEM). The dried-specimen-teasing method appears useful, because of the ease of preparation of the specimens, its reproducibility, and the degree of visibility and preservation of cell surface structures and intraclonal relationships. Our initial observations, using combined EM techniques, indicate that clonal cell topography is highly variable and that this variability appears to be related both to the relative age and proliferative status of the colony. Based on work to date, we suggest that topographical and spatial analysis, in vitro of cloned, agar-embedded hemopoietic stem cells is possible with simple modifications of conventional SEM preparative techniques.


Qualitative and quantitative aspects of labeling cell surface carbohydrates using lectins as probes.

Lectins are proteins which bind mono- and oligosaccharides with great specificity. Many polysaccharides, glycoproteins and glycolipids which are important constituents of cell walls and surface membranes of prokaryotic and eukaryotic cells, contain sugar moieties with which lectins can interact. As a result lectins have been extensively used for the study of cell surface and membrane structure of their labeling. This paper reviews and evaluates the available methods for the preparation of fluorescent, electron-dense and radioactive lectin derivatives. The procedures involved in the visualization of lectin binding under light and electron microscopy are described. In addition, the methods for quantitative evaluation of the fluorescence on labeled cells and for analysis of the number of cell surface lectin binding sites using radioactively-labeled lectins are outlined. Examples are given for the application of fluorescent lectin derivatives for the detection of specific saccharide-containing molecules on the surfaces of living or fixed microbial cells and various normal and neoplastic cells. The use of lectins to demonstrate the dynamic nature of cell membranes and to detect changes in membrane structure or organization which occur or organization which occur during differentiation, development or after neoplastic transformation is discussed.


Hapten-antibody conjugates as probes of the lymphocyte surface.

Various forms and applications of the hapten-sandwich method for labeling cell surface antigens are reviewed. This is a two-layer antibody technique: the first antibody that binds antigen has been chemically modified so that a number of hapten groups are attached; the second antibody is directed against the hapten and has been coupled to a marker for amplified detection. This methodology is particularly suited to circumstances where high sensitivity and specificity are required for labeling alloantigens or for discriminating between antigens in double-labeling procedures. It is applicable to fluorescence visualization, electron microscopy, and radioisotopic labeling. It lends itself also to procedures for specific separation of differing lymphoid cell populations, and has been adapted to studies of cell activation in the immune response.