Serum alpha-feto protein (alpha-FP) and hepatoma in Rhodesian Africans. Alpha-feto protein (alpha-FP) is now accepted as a very useful serum marker of hepatocarcinoma, and since its discovery in 1963, surveys have been carried out in various regions of the world, where hepatoma is particularly prevalent, which have shown that alpha-FP may be detected in 50 to 80% of patients with liver cancer. The sera of 30 Rhodesian African patients with histologically proven hepatoma were tested in this study for the presence of alpha-FP. In only 47% was alpha-FP detected. Thus it is our experience that a positive result is strong evidence in favour of a diagnosis of hepatocarcinoma whilst a negative result is of little value, necessitating that liver biopsy be carried out in all patients with clinical suspicion of liver cancer.


Hepatitis associated antigen, cirrhosis and primary carcinoma of the liver in the Gambia. 45% of 65 Gambian patients with cirrhosis, and 53% of 63 patients with primary liver cancer were found to have hepatitis-associated antigenaemia, re-inforcing the possible aetiological role of hepatitis in these two common diseases of Africa.


Heterologous reactions involving parasites, blood group antibodies and tissue components. The sera of many patients with malaria and filariasis, and also anti-A and B blood group sera, were found to react by immunofluorescence with the somatic musculature of nematodes, especially Ascaris, and human and rat muscle, especially the skeletal type. These reactions were attributed to a polysaccharide related to AB substance in adult nematodes and to raised AB antibodies in malaria. Similar heterologous reactions were found to involve the integument of schistosome adult worms and the microfilariae of Loa loa, which were attributed to the incorporation of host AB blood group substances into the parasite. Other parasites and sera gave mainly negative results. These heterologous reactions constitute a potential hazard in immunofluorescence tests, against which skeletal muscle provides a control. The antibodies concerned were not operative in complement fixation tests, but there was a relationship to anti-complementary activity which suggested the transient presence of a circulating antigen.


Leishmaniasis in Brazil: X. Some observations of intradermal reactions to different trypanosomatid antigens of patients suffering from cutaneous and mucocutaneous leishmaniasis. In spite of the difficulty of antigen standardization, paired intradermal skin tests demonstrated qualitative differences between leishmanin and in vitro exo-antigen. Some of these differences may be due to the destruction of certain antigens by phenol, while others seem to reflect basic antigenic differences. Leishmania mexicana amazonensis in vitro exo-antigen produced immediate anaphylactic reponses in 74.4% of the patients with parasitologically proven cutaneous leishmaniasis. Although more individuals responded to leishmanin, 82.4% as compared to 73.9%, the leishmanial in vitro exo-antigen gave significantly larger delayed responses, Trypanosoma cruzi trypamosomin elicited delayed reactions, while trypanosomal in vitro exo-antigen only produced immediate anaphylactic reactions in persons with active cutaneous leishmaniasis.


Another example of a B variant.

Another example of a B variant. A family in which five members has a weak B variant and one member had a normal A and weak B reaction is reported. It appears that these phenotypes arise by the inheritance of a variant allele of the ABO locus. Their cells react weakly when added to anti-B sera, and they have normal anti-A activity and an absence of anti-B in their sera. Additionally, all of the members tested had increased levels of cellular H substance and all of the affected secretor members of the family secreted normal amounts of both B and H substance. Transferase activity of the sera of these individuals confirmed the presence of the B-gene specific transferase.


A pH-dependent auto-agglutinin with anti-P specificity. An agglutinin with apparent anti-P specificity, reactive only at pH values below 6.0, was found in the plasma of a healthy female blood donor. It is postulated that this agglutinin recognizes latent receptors revealed by structural alterations to red blood cell membranes which occur in an acid environment.


Cross reactivity between H-2K and H-2D products. III. Effect of H-2K-H-2D cross sensitization on skin graft survival. Cross sensitization between the products of the H-2K and H-2D loci has been demonstrated by an experimental design in which recipients were sensitized with tissue bearing determinants controlled by one locus (e.g., H-2K), and then challenged with skin grafts expressing determinants controlled by the other locus (e.g., H-2D). In one combination, experimental grafts were rejected faster than first-set control grafts. Cellular rather than humoral mechanisms appeared to mediate this effect. In a second combination, prolonged survival of experimental grafts were observed. This enhanced survival may have been influenced by serum antibody, either to H-2 or Ia determinants.


Common geography as a clue to causation. Certain diseases characteristic of modern civilization have been shown to have a similar worldwide geographical distribution, and in developing countries a similar socio-economic distribution. This suggests that some causative factor may be common to each, and evidence is provided to support the hypothesis that this might be the over-refining of carbohydrate foods.


Serum immunoglobulin and protein concentrations in Zambian African patients in Lusaka. Serum immunoglobulins IgG, IgA, IgM and IgD and gamma-globulin, total globulin and albumin concentrations have been determined in 136 adult Zambian African hospital in-patients, with several diagnoses, in Lusaka, Zambia. The study gives base-line data for that population. A significant positive correlation between serum IgG and gamma-globulin concentrations (P less than 0.001) suggests that for routine use the less costly method for serum gamma-globulin determination gives equally useful information as that for IgG.


[Spectral and photometric characteristics of different batches of auramine O (OO)]. The spectra of absorption and fluorescence of Auramine O (OO) specimens in water solution and in cells, stained with Schiff-type auramine-SO2 reagents for DNA, were investigated. Using different specimens of the stain the accuracy and sensitivity of the cytofluorometric method were estimated by the next parameters: the value of the useful signal from the nucleus (no), the value of the production of no by the time constant of fading (tau), the value of proportion of the signal to the background (no/nphi) and, in addition, the value of the variation coefficient under the measurement of the rat liver tetraploid cells. The purity of the dye was shown to interfere largely with accuracy and sensitivity of the above method. Auramine 00 of the Reanal production appeared to have the best photometricl characteristics. For this, the maxima of the absorption spectrum in the 320--700 nm region in the solution and in the cell were 371 and 433 nm, and 375 and 436 nm, resp. The maximum of the fluorescence spectrum was found 521 nm and 526 nm, resp. for the solution (0.02%) and for the Auramine-SO2 treated cells. Moreover, an isomer of Auramine O (00) -- Auramine G can be used for detection and photometrical measurement of DNA and glycogen after the Feulgen and PAS reactions, resp.


[Method for the morphological identification of individual brain neurons intravitally and after fixation]. Pyramidal neurons stained with methylen blue (from layers III and V of the cat brain cortex) are revealed in vivo after a special preparation of the contact objectives in the reflecting light. Small pieces of the brain tissue with identified neurons were separated under visual control, after which an oriented piece placed in a special grid-basket appeared to be stabilized by the borders of the grid. This method permits investigating the same neurons after fixation by means of histological light or electron microscope techniques.


[Alpha-fetoprotein in assessing obstetric diagnosis (author's transl)]. Radio-immunologic measurement of alpha-fetoprotein is possible in small amounts of blood and amniotic fluid. During the first half of pregnancy the AFP values in maternal serum are lowered when there is fetal abnormality (hydatid cyst, immnent abortion). In the second half of pregnancy high values of AFP also indicate danger to the fetus (Rh-incompatibility). The older the pregnancy the AFP values in amniotic fluid fall. In pathologic pregnancy (hydramnion, anencephaly, trisomy) AFP concentration was normal or raised. Measurement of the AFP level in umbilical cord -- blood provides further diagnostic chances: assessment of the duration of pregnancy and, therefore, of fetal maturity.


The soluble antigens of Rickettsia prowazeki, R. typhi and R. canada. Investigation of their interrelationship by various serological methods. The purpose of this research is the isolation of an eventual species-specific fraction from the "soluble antigen" of Rickettsiae. The "soluble antigen" of R. prowazeki (Breinl strain), R. typhi (Wilmington strain) and R. canada were purified at 25% saturation with ammonium sulphate (PSA). Corresponding antisera were produced in rabbits. The serological methods used were the complement fixation, the micro-agglutination, the precipitation method in capillary tubes and the immuno-diffusion method carried out with complete and previously cross-absorbed antisera. Furthermore, the PSA were subjected to immuno-electrophoretic and disc electrophoretic fractionation. The PSA of R. prowazeki was found to contain at least 4 group-specific fractions. A species-specific component could be demonstrated with certainty only with the precipitation method in capillary tubes carried out with previously cross-absorbed antisera. The PSA of R. typhi contains 5 fractions of which 4 are group-specific and one is species-specific. This result was confirmed by all methods. The PSA of R. canada: the maximum of 3 components could be detected with the help of immuno-electrophoretic fractionation. A fourth antigenic determinant was made apparent by the presence of corresponding antibodies in the anti-R. canada PSA only.


Immunological activity of the peptidoglycan. Studies on the immunology of peptidoglycan received impetus from the initial observation that the rabbit Group A-variant streptococcal antisera were a rich source of antibodies to peptidoglycan. Indeed, quantitative precipitin studies revealed concentrations as high as 7-10 mg/ml of antiserum. The amount of antibody after Group A-variant streptococcal immunization is much greater than the amount in the sera following immunization of rabbits with the Group A or C streptococci. Furthermore, earlier studies had shown that the purified peptidoglycan obtained as a residue following extraction of streptococcal cell walls with hot formamide was a poor antigen. Both the hexosamine polymer and the peptide moiety are antigenic. Use of the solid phase synthesized pentapeptide L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala and related similar peptides facilitated the determination of the fine structure of the immunodominant site of pentapeptide. The evidence points to the C-terminal D-Ala-D-Ala as the immunodominant determinant. Because of the similarity of the peptidoglycans of a number of different bacteria, it would be anticipated that they would cross-react immunologically, and this has been shown to be the case. The biological and medical significance of antibodies to peptidoglycan has yet to be determined. Certainly the exposure to this ubiquitous substance which occurs in all the indigenous bacteria of the respiratory and the gastrointestinal tract must mean that from an early age and through advancing years there is a constant stimulation of the immune response to peptidoglycan. Because the immunochemistry of these substances is now firmly established, there is a scientific basis for proceeding with the medical and biological implications of peptidoglycan immunity.


Measurement of peptidoglycan antibodies by a radioimmunoassay. Staphylococcus epidermidis peptidoglycans solubilized by sonication or lysozyme digestion, and synthetic peptidoglycan analogs such as HSA-carboxymethyl-Gly-L-Ala-L-Ala-D-Ala-D-Ala (HSA-pentapeptide) or L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala (pentapeptide) have been labeled with 125I and tested for their applicability in the radioactive antigen binding assay. Use of radioiodinated Staph. epidermidis peptidoglycans was found to be considerably impeded by the presence of at least 2 different antigenic sites on such molecules, the pentapeptide and the glycan determinant. Application of labeled HSA-pentapeptide was limited by the necessity to use PEG for precipitation of Ag-Ab-complexes and by short linear potions of binding curves. However, the synthetic pentapeptide hapten, radioiodinated by the active ester method of BOLTON and HUNTER, proved to be a most useful regent for the selective measurement of pentapeptide antibody. Inhibition studies indicated that the immunological specificity of the labeled hapten was retained. Pentapeptide binding curves were linear from 15-500 g/ml of antibody. Generally, there was good agreement between pentapeptide antibody concentrations measured by radioimmunoassay and quantitative precipitation.


Isolation and characterization of homogenous rabbit antibodies to Micrococcus lysodeikticus with specificity to the peptidoglycan and to the glucose-N-acetylaminomannuronic acid polymer. The antibody response of rabbits to Micrococcus lysodeikticus is characterized by the production of a high concentration of antibodies which manifest markedly reduced heterogenicity. The specificity of these antibodies was studied and it revealed that M. lysodeikticus contains 2 major antigens: both the glucose-N-acetyl-aminomannuronic acid polymer obtained by formamide extraction of the cell walls and peptidoglycan solubilized by ultrasonic treatment gave precipitin reactions with hyperimmune antisera. By means of inhibition studies of the glucose-mannose polymer specificity, glucose appeared as the immunodominant sugar in the majority of antibodies studied. Inhibitions studies also confirmed that both the glycan and peptide moieties constitute antigenic determinants of M. lysodeikticus peptidoglycan. Antibodies to the glucose-mannose-polymer and the peptidoglycan were specifically fractionated by use of immunoadsorbents formed from lysozyme solubilized cell walls and activated Sepharose. Both antibody specificities showed a limited heterogeneity by isoelectric focusing. Finally, because antisera to M. lysodeikticus are a rich source of antibodies to peptidoglycan, emphasis is placed on the possible usefulness of this system for studies of clonal dominance.


[A contribution to the morphology of Sudeck's syndrome (author's transl)]. Histologic illustrations of stages of Sudeck's syndrome in the case of a 22 year old man, following amputation of the left mid-foot. Histometric techniques were also used.


[Serological studies on Staph. aureus strains from various processes of a disease of human origin (author's transl)]. This paper deals with serological studies on 332 Staph. aureus strains of human origin having been isolated from various sources. On the basis of typing with 17 factor sera, a great number of groups and types was obtained occurring among the strains of several processes of a disease without showing special serological structural formulae which would have been typical for pathological changes of certain localisation. But still deviations have been observed between the strains of most of the series of observation which proved to be significant differences of incidence and referred to several groups and types. In this isolated observation of the single factors, different antigen-structures occurred as result.


[Neuraminidase neutralizing antibodies in pigs with chronic Erysipelothrix rhusiopathiae infection (author's transl)]. Investigating sera from pigs with chronic E. rhusiopathiae infection relative gamma-globulin values, agglutination titers, and neuraminidase-neutralizing humoral antibodies were determined at the 12th and 15th, 11th and 14th, and 3rd and 6th month after infection. We found increasing gammaglobulin values and increasing neuraminidase neutralizing antibodies but decreasing agglutination titers. Few animals don't have antibodies against neuraminidase and died during the observation period. Furthermore immunodiffusion test demonstrated precipitating lines of antibodies against living E. rhusiopathiae cells and active neuraminidase. No or very weak bands were observed against heat killed cells and heat-inactivated enzyme.


Studies on serotyping of Neisseria gonorrhoeae. By means of an agar-gel double diffusion technique the existence of five serological types of Neisseria gonorrhoeae was revealed by demonstrating the antigens responsible for type specificity. Based on these findings a typing method has been developed using a microprecipitin technique in agar-gel. 381 gonococcal strains have so far been typed by means of five cross-absorbed rabbit antisera.


[Kanner's syndrome and childhood schizophrenia]. The examination of 32 children with Kanner's syndrome of early infantile autism permits to assume that this syndrome in some of the cases is expressed only by inborn anomalies which correspond to constitutional psychopathy in adults. In most of the cases this syndrome forms the initial expression of child schizophrenia. In separate cases disorders very similar to Kanner's syndrome may be seen after the first olliterated attack during early childhood (up to 3 years). A comparative study of the same indices of development of 268 children with an early onset of schizophrenic process in spite of some differences confirms that Kanner's syndrome is very close to childhood schizophrenia. An analysis of genealogical data shows genetical relations of Kanner's syndrome with child schizophrenia.


Studies on topical antiperspirant control of axillary hyperhidrosis. Axillary hyperhidrotics is reviewed from the standpoint of anatomical factors, physiological mechanisms and the history of methods of control. Anhydrous aluminum chloride and anhydrous zirconium tetrachloride are shown to be superior topical agents for partial control of axillary sweating when applied as a powder or in anhydrous nonreactive vehicles. Complete anhidrosis as demonstrated by sustained garment armpit dryness could be achieved in hyperhidrotics within 48 hours by the following trinary antiperspirant system: (1) a saturated solution of aluminum chloride hexahydrate or zirconyl chloride in absolute ethanol or isopropyl alcohol, (2) application to the dry axilla at times of sleep or other prolonged non-sweating period, (3) water vapor occlusion of area for 6 to 8 hours by means of Saran wrap. The hypothesis is presented that metallic antiperspirants act by reflux entrance into the terminal intraepidermal eccrine duct, slowly combining with the intraductal keratin, to produce a fibrillar contraction (super contraction) of keratin and hence functional closure, not histologically evident. This altered keratin is shed weeks later, with the consequent return of ductal patency and sweating.


Development of high intensity narrow-band lamps and studies of the irradiation effect on human skin. Irradiation with high intensity lamps. A compact light source has been developed. It consists of a high-pressure mercury lamp, a shutter, and water-cooled filters for selected wavelength regions. A mixing device permits simultaneous irradiation from two lamps of different wavelengths. The spectral distribution of the light for seven filter combinations and the construction of a power meter are also described. The application of the lamps to clinical investigations is illustrated by determination of minimal erythemal and blister doses, as well as pigmentation, for various groups of patients. The light intensities available are high enough to make pain threshold measurements possible in the UV and visible regions.


Products of epidermal protein synthesis. The products of epidermal protein synthesis in tissue slice and cell-free systems have been characterized by chromatographic, electrophoretic, and immunologic methods. Radiolabeled proteins of heterogeneous size are found in the supernatant and particulate fractions from both systems. The major labeled proteins in the supernatant fraction have molecular weights of approximately 43 000 and 65 000. The natively-insoluble proteins of the particulate fraction were solubilized either with desoxycholate or alkaline urea. Subsequent analysis of theses fractions yielded a labeled, high molecular weight, urea-soluble protein whose electrophoretic mobility was slower than that of the heavy chain of myosin. The immunologic reaction between the radiolabeled fraction and antibody raised against filamentous protein suggests that synthesis of proteins identical to epidermal filamentous protein occurs in the in vitro systems.


Electron microscopic study of nevic corpuscle. An intradermal nevus, in which a number of nevic corpuscles were clearly observed, was studied, using the electron microscope. Nevic corpuscles contained laminated cells consisting of flattened cytoplasmic processes stretching across the corpuscles to form a complicated labyrinth. The perikaryon of these cells contained premelonosome-like dense bodies and other organelles including mitochondria, rough-surfaced endoplasmic reticulum, free ribosomes, and Golgi apparatus. Neither axons nor dendrites were found in these areas. From these observations, it was concluded that the nevic corpuscles were composed exclusively of nevus cells and could be clearly distinguished from Meissner corpuscles. This view would support the idea of a unitary origin of nevus cells. In addition, an isolated cilium found in a laminated cell is briefly discribed.


Liver biopsy in methotrexate-treated psoriatics-a re-evalution. Two-hundred and eighty-six liver biopsies were performed in 139 psoriatics on treatment or considered for treatment with methotrexate. In 56 psoriatics included in this study both pre- and post-methotrexate biopsies were performed, the average methotrexate dose being 936 mg. None of the data showed statistically significant differences between pre- and post-methotrexate biopsies, with the exception of an increase in fattly infiltration, found when comparing all pre-methotrexate biopsies with the total number of latest post-methotrexate samples. As expected, alcohol seemed to be significantly associated with liver fibrosis in pre-methotrexate biopsies. An patients, although potassium arsenite alone has not been proved to be the cause of liver damage among psoriatics included in this study. While only 1 of 22 psoriatics with a total normal biopsy had been on arsenite, 6 of 18 of the same group of psoriatics who had fibrosis had been on this drug earlier. Although no statistically significant differences related to fibrosis and cirrhosis could that in three cases liver cirrhosis did appear in a biopsy from a methotrexate-treated psoriatic who had signs of fibrosis of cirrhosis in a pre-methotrexate biopsy. This incidence is low in relation to the total number of patients treated. The relatively low incidence of cirrhosis found in the present study, as in earlier studies by our group is believed to be due to the use of an intermittent dosage schedule. The study showed that early fibrosis and cirrhosis seem to appear, with very minor abnormalities in laboratory results. This finding indicates the necessity of performing liver biopsies in the control of psoriatics on long-term methotrexate therapy. The difference between biopsies from psoriatics and liver biopsies from control patients may indicate that severity of disease may be a complicating factor in the pathogenesis of the liver damage.


HL-A antigens (17, 27, UPS) in psoriasis with special reference to patients with arthritic lesions. Fifty patients with longstanding and severe psoriasis vulgaris have been tissue-typed. HL-A 17 and 27 from second sublocus and UPS from third sublocus have been found to occur in increased frequency. HL-A 27 was found to be more prevalent among those with arthritic lesions. The lack of significantly increased frequency of HL-A 13 is discussed.


Circulating immune complexes in a patient with prolonged gonococcal septicemia. A case of prolonged gonococcal speticemia is presented. Lack of genital symptoms, of demonstrable gonococcal antibodies in whole serum, and initially also of skin lesions contributed to a delayed diagnosis that was finally confirmed by positive blood culture. Evidence of circulating immune complexes was obtained by the demonstration of gonococcal IgG antibodies after dissociation of serum by gradient centrifugation of pH 4.0. The possibility that circulating immune complexes may contribute to the symptomatology of gonococcal septicemia is pointed out.


Anti-basement membrane antibodies in sera from patients without bullous pemphigoid. Sera from 208 individuals were examined for antibasement membrane (anti-BM) antibodies by means of an indirect immunofluorescence technique. In sera from patients with bullous pemphigoid 19 of 25 had anti-BM antibodies in titres varying from 80 to 32 000. In 2 of 42 sera from patients with psoriasis, anti-BM antibodies were demonstrated (titres 40-640). Of 58 sera from patients with leg ulcers, antibodies were found in 4 (titres 40-320). Antibodies were found in 2 of 43 patients with cardio-vascular disease (titre 80) and in 1 of 40 control persons (titre 40). Serum from one of the patients with leg ulcers caused a punctate staining pattern unlike the tubular pattern seen in all other positive sera.


Epidermal intercellular binding of concanavalin a and pemphigus antibody. The binding sites of Concanavalin A (Con A) and pemphigus antibody in the intercellular spaces of the human epidermis have been investigated. Extraction of human skin with phosphate-buffered saline (PBS) and 70% ethanol resulted in complete loss of the binding sites of the pemphigus antibody, whereas the extracted skin still retained its reactivity to Con A. Preincubation with more than 0.25 mg/ml of Con A with normal skin caused a reduction in the binding of pemphigus sera. However, preincubation of pemphigus sera with human skin caused no inhibition of the subsequent binding of fluorescein-labelled Con A. These results indicate that Con A binding sites might be related to, but not identical with those of pemphigus antibody.


The effects of X-ray, ultraviolet and infrared irradiation on the basement membrane zone antibody reaction of the human skin in vitro. Two patients with bullous pemphigoid were treated with radio cobalt or betatron irradiation. Thereafter bullous eruptions developed in the non-irradiated skin. It is known from the literature that ultraviolet light applied to the normal-appearing skin of patients with bullous pemphigoid produced fresh histologic and immunohistochemical changes in the irradiated skin. X-ray irradiation applied to human skin biopsy specimens using single doses of 7 000 and 8 000 R resulted in an increase of basement membrane zone antibody binding of 2-3 titre dilution steps. An increase of 3-4 titre dilution steps was achieved by irradiation with ultraviolet light. Infrared irradiation of skin biopsy specimens, on the contrary, caused the total loss of the basement membrane zone fluorescence. It merits further study, whether these contrary effects on the basement membrane zone antibody binding of human skin in vitro (activation of bound antigen after X-ray or ultraviolet irradiation and inactivation of antigen caused by infrared irradiation) are of clinical value concerning the bullous pemphigoid problem in vivo.


Poikiloderma congenitale-an early case of rothmund-thomson's syndrome. A case of congenital poikiloderma is presented. Two types of congenital poikiloderma can be differentiated on the basis of onset of symptoms and configuration of skin lesions, i.e. Rothmund-Thomson's type and Wodniansky's type. The case presented here is of the Rothmund-Thomson's type, but showed only minor abnormalities apart from skin changes and hair loss. A slightly elevated concentration of lysine and cystine was found in the urine of our patient.


Surgical treatment of paronychia granulomatosa hallucis. Paronychia granulomatosa is a complication incident to unguis incarnatus; surgery for this complaint was performed on 26 patients with 52 granulomas. The operation was first described by Bartlett in 1937. The postoperative hospitalization was 3 days and the patients returned a week later to have the sutures removed. After the operation, ten remaining granulomas were observed of which 6 were cured by operation and 2 conservatively, leaving 2 under continuing conservative treatment (5 months and 1 month, respectively).


Normal response to thyrotrophin releasing hormone (TRH) in familial thyroxine-binding globulin deficiency. The response in serum thyrotrophin (TSH) to thyrotrophin releasing hormone (TRH) has been studied in 5 euthyroid patients with familial thyroxine-binding globulin (TBG) deficiency. Total serum thyroxine (T4), serum triiodothyronine (T3) and free T4 index and free T3 index were significantly and equally decreased, but in spite of these findings the serum TSH and response to TRH was normal. The TRH test seems to be a better indicator of the euthyroid state in familial TBG deficiency than the measurement of free T4 and free T3 in serum.


Cellular localization of somatostatin in endocrine-like cells and neurons of the rat with special references to the A1-cells of the pancreatic islets and to the hypothalamus. With the indirect immunofluorescence technique somatostatin, a recently isolated and structurally characterized hypothalamic tetradecapeptide, which inhibits growth hormone release, has been traced in different tissues of the rat. Somatostatin or somatostatin-like immunoreactivity (SLI) was present in both neurons and endocrine-like cells. Somatostatin positive nerve cell bodies were observed in the periventricular region in the anterior parts of the hypothalamus and in probable nerve endings in the external and internal layer of the median eminence and in the pituitary stalk and in the ventromedial, arcuate and ventral premammillary nuclei and to a small extent in the periventricular region. Somatostatin positive nerve fibers were also found in the posterior pituitary, indicating the existence of a third neurosecretory hypothalamo-hypophysial system, and in nerves in different layers of the wall of the small and large intestine. These nerves may possibly be of sensory nature since some cell bodies of the spinal ganglia showed a weak immunofluorescence. Somatostatin or SLI was found in certain cells localized in the periphery of the pancreatic islets. In parallel studies with glucagon antibodies it could be established that the somatostatin positive cells and the glucagon positive cells were not identical but localized extremely close to each other. Furthermore, with the Hellman-Hellerström silver staining technique it could be shown that virtually all somatostatin cells are argyrophilic and vice versa. Somatostatin positive cells thus constitute the socalled A1-cells (D-cells). Glucagon positive cells, on the other hand, do in all probability not exhibit argyrophilia. The occurrence of probable somatostatin containing cells in the pancreatic islets is of special interest in view of several recent studies demonstrating an inhibitory action of this peptide on glucagon and insulin secretion. Somatostatin or SLI was found in a rather small number of cells in the thyroid gland with a parafollicular position. These cells exhibited a positive immunofluorescence also after pretreatment of the antiserum with calcitonin. Furthermore, endocrine-like cells in the stomach and in the intestine were also somatostatin positive. A large number of cells in various tissues, especially in the lamina propria of the gastro-intestinal tract, showed a strong fluorescence both after incubation with control serum and with FITC conjugated serum alone. The present results indicate that somatostatin or a somatostatin-like peptide(s), in addition to its inhibitory action on growth hormone release, may play a physiological role at many other levels of the organism.


Histochemical analysis of epidermal cells and mucous cells in the skin of Torpedo ocellata Rafinesque. The histochemical analysis carried out on the mucus of the epidermis of Torpedo ocellata RAF. showed that it is not only produced in considerable quantities by the mucous cells but is also elaborated by the epidermal cells of the skin. The mucus elaborated by the mucous cells is rich in carboxyl and phosphate containing MPS and in sialic acid containing glycoproteins. It also contains a neutral mucopolysaccharide fraction relatable to glycogen and a considerable quantity of chondroitinsulfates B. Epidermal cells produce a secretion with the same histochemical characteristics as the mucus elaborated by the mucous cells and contain mucopolysaccharide-protein complexes. Their secretory activity is also discussed in relation to their functionality.


Light microscopic differential staining of epoxy-embedded adenohypophysis. Light microscopic histochemical methods for differential staining of adenohypophysis glandular cells were applied to Epon-embedded tissue, after prior softening of resin with saturated solution of NaOH in absolute ethanol. All investigated staining procedures, i.e. carmoisin L-orange G, PAS-orange G, PAS-range G-methyl blue, Gabe's aldehyde fuchsin-Halmi counterstain, aldehyde thionin-PAS-orange G and performic acid-alcian blue-PAS-orange G were found to give satisfactory results and tissue structure showed no signs of damage caused by the Epon-dissolving reagent. The features of light microscopic image given by plastic-embedded semithin sections: nearly 2-dimensional picture as well as the highest possible resolving power provide the opportunity for much more precise and detailed examination of adenohypophysis prepared that way, as compared with conventionally paraffin-embedded gland. Moreover, the described technique permits to establish a direct connection between light microscopic differential histochemistry of adenohypophysis and the electron microscopy by means of adjacent sections.


Investigation of the presence of chromaffin tissue in the ovary of the percoid teleost fish, Nandus. The ovary of Nandus shows chromaffin tissue at different sites below the tunica albuginea, along the ovigerous lamellae and near the oocytes. The tissue can be mistaken for interstitial cells as it shows intense sudanophilia. That it is chromaffin tissue is confirmed by its positive reaction to all the usual histochemical tests for chromaffin. This tissue may represent some stage in the development of "corpora lutea".


The effect of ACTH and cortisone on basement membrane in relation to the embryological origin of the epithelium. The changes of intensity of PAS reaction in basement membranes of epithelial cells derived from 3 different germ layers in rats after treatment with ACTH and cortisone were observed. After treatment with cortisone an increase of PAS reaction in all investigated basement membranes was observed. Influence of cortisone on all basement membranes was independent of the embryological origin. The results after treatment with ACTH were not uniform.


[On the possibility to demonstrate the presence of catecholamines in enterochromaffin cells: critical review (author's transl)]. Various data contributing to support, after Lillie et al., the presence of catecholamines instead of (or in addition to) 5-hydroxytryptamine in the enterochromaffin cells, are submitted to analytical criticism by the Author. Topics of the discussion are mainly the behaviour of the diazo-reaction (in acid medium), the fluorescence induced by formaldehyde, and various reactions carried out either directly or following a chemical block. The Author demonstrates that 5-HT is present in the enterochromaffin cells and that it is impossible, at present time, to prove the presence of catecholamines in these cells on the basis of histochemical data. The Author debates also how biochemical or pharmacological tests may supply the lacking of histochemical proofs in demonstrating the presence of catecholamines; it is possible by applying these procedures to study situations like those postulated by Lillie et al., extending a comparative research also to other elements of the enterochromaffin cell system after ERSPAMER; the examples here reported however do not agree perfectly in some of the cases with these views.


Lysosomal hydrolases in the trigeminal ganglion of mice afflicted with an hereditary sensory neuropathy (dystonia musculorum). Selected lysosomal hydrolases have been investigated in the trigeminal ganglion of mice afflicted with an hereditary sensory neuropathy (dystonia musculorum). This was done using direct enzyme histochemistry. Correlative electron microscopy was also used to further elucidate perikaryal changes. The earlies observed lesion in the trigeminal ganglion of afflicted mice was numerous axon swellings containing intense lysosomal hydrolase activity. Subsequent to this observation, numerous neurones showed central chromatolysis, eccentric nucleus and increased lysosomal hydrolase activity. As various neurones throughout the ganglion underwent the classical chromatolytic reaction, the Golgi apparatus moved to a juxtanuclear location, and there was a focal juxtanuclear accumulation of lysosomes. During the later stages of the disease, a striking decrease in neuronal hydrolase activity characteristic of neuronal atrophy was observed. These results are consistent with earlier suggestions that loss of sensation in the disease could be due to an interruption of axonal transport in primary sensory of neurones.


[The so-called filaments in lafora-bodies, corpora amylacea and Bielschowsky-bodies. Fasciolar substructures in accumulated intraplasmatic material of cellular dysmetabolies (author's transl)]. Both the corpora amylacea and Lafora bodies have been described in the literature as filamentous structures. On electron microscopic examination they are often composed of ribbon-like fasciolar units. The ultrastructural appearance of these structures is discussed on the example of an amylopectin model. These fasciolar substructures are also encountered in Bielschowsky bodies and are then considered to be a sign of intracellular accumulation of dysmetabolic products. The histological and electron microscopical similarity of corpora amylacea, Lafora bodies and Bielschowsky bodies suggest that all these structures are an unspecific, nearly identical endproduct of various intracellular metabolic disturbances. The pathognomonic significance of these structures depends on their regional distribution: the common corpora amylacea occur predominantly in the astroglia, the Lafora bodies typical in the neurons and the Bielschowsky bodies are restricted to the neurons of the exterior pallidum.


The diagnosis of chronic simple conjunctivitis. Vital staining of tarsus with tetrazolium - alcian blue mixture. Vital staining with a tetrazolium - alcian blue mixture gave a characteristic punctate red staining of the superior or the inferior tarsus in 69% of all examined patients with chronic simple conjuctivitis (69 patients subjected to 168 examinations). The superior tarsus was more frequently stained than the inferior (61% against 33%). The staining was most often concentrated centrally or in the middle anterior part of the superior tarsus. If located on the inferior tarsus, it was most often found medially, anteriorly. No corresponding staining was seen in normal eyes. The differential diagnosis involving keratoconjunctivitis sicca and pemphigoid is mentioned. The staining is due to enzymatic reduction of tetrazolium in the cytoplasm of the tarsal epithelial cells. The positive vital staining finding is consistent with a diagnosis of chronic simple conjunctivitis.


Variations of protease inhibitors in foetuses, newborn infants and in some neonatal disorders. Low levels of protease inhibitors have been found on the 1st day of life in IRDS infants. 19 IRDS infants were studied together with foetuses and control term and preterm infants. Alpha1-antitrypsin, antichymotrypsin and alpha2-macroglobulin were measured with the electroimmuno assay. IRDS infants had significantly reduced concentration of alpha-antitrypsin and antichymotrypsin on the 1st day, the level increasing to normal on the 2nd day. In foetuses alpha1-antitrypsin was normal, antichymotrypsin 2% and alpha2-macroglobulin 1/3 of the normal adult level. The protease inhibitors are increased in infants born after premature rupture of foetal membranes. The part, if any, played by protease inhibitors is not entirely understood. The inhibitors may, theoretically, be of some importance in the dissolution of the hyaline membranes, protect against pulmonary vasoconstriction, protect pulmonary tissue against leucocyte and macrophage proteolytic enzymes and inhibit the release of or counteract vasoactive substances that might take part in the development of shock in IRDS babies.


Bacterial antigen and acid phosphatase in macrophages in experimental pyelonephritis. Experimental retrograde E. coli pyelonephritis was produced in rats. The study covered the period from 6-24 hours up to 6 months. Macrophages in the renal tissue were studied using immunofluorescence staining for bacterial E. coli antigen and histochemical staining for aicd phosphatase. A comparison of sections stained according to the two methods showed that antigen-containing macrophages in nearly all cases yielded a positive reaction for acid phosphatase. On the other hand, in several kidneys acid phosphatase-positive macrophages occurred which in consecutive sections studied by immunofluorescence did not contain antigen. The possibility of using staining for acid phosphatase as a screening method for the detection of active, antigen-containing macrophages in human chronic pyelonephritis is discussed.


Canine mycoplasmas: serological studies of type and reference strains, with a proposal for the new species, Mycoplasma opalescens. The type and reference strains of all Mycoplasma species or "serogroups" presently known to occur in dogs were compared serologically. Tests of growth inhibition and metabolism inhibition as well as indirect immunofluorescence tests did not disclose any cross reactions. Examination by double immunodiffusion revealed at least one antigen specific for each strain tested. In addition, the common antigens were found within the glucose fermenting group and within the arginine metabolizing group of Mycoplasma strains, respectively. No antigens were shared by these groups. The arginine positive strain MH5408, serogroup D was found to differ serologically from all other arginine positive Mycoplasma species. Serogroup D is therefore a new species for which the name Mycoplasma opalescens is proposed. The type strain is MH5408 (ATCC 27921 and NTCC 10149). Finally, serological data are presented which relate strain HRC689 to M. mycoides.


O-antigenic specificity of lipopolysaccharides from Bacteroides fragilis ss. fragilis. Lipopolysaccharides (LPS) isolated from three strains of Bacteroides fragilis ss. fragilis were examined by indirect haemagglutination and inhibition of haemagglutination in unabsorbed and absorbed rabbit antisera. Six different antigenic specificities were detected. All serological activity was destroyed by oxidation of LPS with periodate.


Local tissue effects of surface-applied ENT drugs. Local tissue effects caused by a selected group of topically applied ENT-drugs are analysed. By means of a testing system consisting of vital mircoscopy, infrared thermography and microangiography applied to hamsters and rabbits, an evaluation of the varying degree of tissue injury is made. According to the degree of tissue damage the tested substance could be graded in three groups (I-III). The drugs belonging to group I show fairly little microcirculatory disturbance, while those belonging to group III cause tissue necrosis. Our conclusion is that topically applied drugs should be used with caution, especially on previously injured tissues.


Quantification of the inhibitory effect of eriochrome black and sodium nitrite on non-specific immunofluorescent staining. To evaluate non-specific staining (NSS) of animal tissues by FITC-labelled immunoglobulins a model system was developed. HeLa cells were treated with labelled antisalmonella globulins and the fluorescence intensity of the cells was determined quantitatively by means of a fibre optic probe system. This system was used to determine the optimal treatment conditions (adequate concentrations, duration of treatment) using the two NSS-reducing agents eriochrome black and sodium nitrite. Approximately the same inhibitory effects (40-50 per cent using nitrite; about 85 per cent using eriochrome black) were obtained by conjugates of different F/P ratio values. The fibre optic probe system was also used to determine the effects of the above-mentioned agents on the NSS of liver sections. In this system, the NSS was strongly reduced by eriochrome black whereas nitrite treatment did not induce any inhibitory effect. The applicability of nitrite and eriochrome black as NSS-reducing agents was further demonstrated by the fact that they had no influence on the specific fluorescence intensity of salmonella bacteria. The effect of eriochrome black was also studied in clinical specimens infected with salmonella or tularemia bacteria.


In vitro and in vivo release of histamine by contrast media in the rat. Some contrast media are able to release histamine from isolated mast cells of the rat and produce degranulation of these cells. Such properties are fairly correlated with the ability to increase vascular permeability in vivo and to produce a drop in blood pressure. If histamine is similarly being released in humans many untoward effects of these agents referred to clinically might be thus explained.


The effect of arfonad on blood proteins and nitrogen components in experimental shock. In experiments performed on anaesthetized dogs the effects of dye shock and peptone shock on changes in serum osmolality, haematocrit value, amino acid level in serum and erythrocytes, and serum total protein and protein fraction levels were determined. In further experid (0.1 mg/kg). It was found that Arfonad exerts an inhibitory effect on the shock-induced reduction in the serum osmolality, plasma volume and on the albumin level.


Thyroxine utilization rate in sucking pigs. In 10 suckling pigs age 28--34 days, thyroxine utilization rate (TUR) was determined by means of the isotope dilution procedure. For this purpose biological half-life of T4 was estimated during 96 h of blood sampling and serum T4 concentration was measured directly by a competitive protein-binding technique. High rate of the peripheral hormonal-iodine metabolism (from 10.3 +/- 0.6 to 21.2 +/- 1.2 mug T4/kg/day) was found in the one-month old pigs. TUR value calculated for these pigs corresponds to that reported in human infants and it is much higher than in the growing ruminants.


The Di Guglielmo syndrome: a study of 17 cases. II. Periodic-acid schiff staining of the erythroblasts. In 17 patients with Di Guglielmo's syndrome the PAS-stained erythroblasts in bone marrow smears have been assessed by a scoring system. Most of the PAS scores were either moderately high or very high--much higher than those normally seen in disorders with benign erythropoietic hyperplasia. On the other hand, very low scores were obtained in a few cases. Whereas a high PAS score always lends strong support to a diagnosis of Di Guglielmo's syndrome, a low score, or even a negative reaction, does not exclude this diagnosis. The highest scores were obtained in cases with many pathological erythroblasts. Staining after diastase digestion confirmed that the positive PAS reaction is due to glycogen. In this test, experience showed that a period of at least 2 hours is required for complete digestion of the glycogen.


The mast cells of the newborn rat diaphragm and their response to histamine liberators. The mast cell population of rat diaphragm was estimated between birth and adulthood and found to rise with an increase in the age of rat studied. Degranulation of these cells was observed in rats from all age groups, following treatment with compound 48/80 and dextran. The association of mast cells with the blood vessel wall in adult rat diaphragm was not observed in the comparable tissues of newborn rats. These findings are discussed in relationship to the poor vascular permeability reactions exhibited by newborn and young rats.


The effect of Trimepranol on thrombocyte function and histamin release in the rat. Trimepranol a beta adrenergic blocking drug released serotonin from rat isolated and plasma-rich thrombocytes in vitro. The release was time- and dose-dependent. With the same concentrations this drug inhibited the histamine release from isolated rat mast cells. Aggregation of isolated or plasma-rich thrombocytes induced by ADP was inhibited by Trimepranol. The inhibition was dose, time dependent and reversed by calcium ions.


On fibrinolysis in diabetes mellitus.

On fibrinolysis in diabetes mellitus. In diabetics, irrespective of age at onset and standing of the disease, the fibrinolytic activity in the blood is decreased, most markedly in obese patients and in those with angiopathy. In this investigation we found--significantly more often than in controls--a low release of the plasminogen activator to the blood from the endothelial cells, and also in one fourth of the diabetics an abnormally low activator activity of the vessel walls. These changes, as well as the significantly elevated levels of inhibitors of the fibrinolytic activity, will impair removal of fibrin deposits and possibly contribute to the development of vascular lesions.


Treatment of Whipple's disease with sulphamethoxazole-trimethoprim. A patient with Whipple's disease responding rapidly to sulphamethoxazole-trimethoprim is described. In spite of initial folate deficiency, the drug was well tolerated for one year. It is proposed that a sulphonamide combined with trimethoprim should be the drug of first choice in the treatment of Whipple's disease. Repeated jejunal biopsy is suggested as the best way to record the effectiveness of treatment.


Immunological activity against tick-borne encephalitis virus in fractions of sera from goats of different ages. A comparative analysis of the antihaemagglutinating, virus-neutralizing and complement-fixing activities of 19 S, 7 S, 4S and 1.5 S fractions of blood sera from goats of different ages pastured in a tick-borne encephalitis focus was carried out. Changes in the immunological activity of different fractions in relation to the age of the animals are discussed.


Tricuspid atresia: clinical course in 101 patients. The clinical profile and course of 101 patients with tricuspid atresia seen at the Children's Hospital Medical Center, Boston, were reviewed; the electrocardiograms, hemodynamic data, associated anomalies, complications of the malformation and surgical results were outlined and a definition of anatomic types and radiologic groups formed in order to facilitate therapeutic decisions. Overall survival to 15 years of age was approximately 50 percent. Tricuspid atresia type 1 (normally related great arteries), group A (decreased pulmonary vascularity) was the most frequent category (54 percent); without surgery, these patients had only a 10 percent chance of survival beyond the 1st year of life. Surgical intervention improved chance of survival to 15 years of age to 50 percent. Overall surgical mortality was 23 percent. The Waterston shunt is the procedure of choice in symptomatic small infants with diminished pulmonary flow (group A). Type II patients with increased pulmonary blood flow (group B) would benefit from a pulmonary arterial band. In contrast, patients with group B, type I atresia would rarely need a pulmonary arterial band. Patients with spontaneously changing hemodynamics (group C) usually require a shunt later in life than those in group A, with more favorable operative results. Our experience indicates the continued need of early surgical palliation. New surgical approaches, such as the right atrial-pulmonary arterial anastomosis, may result in a more prolonged survival.


Relationship of maternal amino acids and proteins to fetal growth and mental development. The relationship of maternal amino acids, proteins, and certain other factors to fetal development (weight and cranial volume at birth and mental and motor scores at 8 months of age) was investigated in 129 gravidas. Maternal prepregnancy weight was significantly related to birth weight and cranial volume in the newborn. Among amino acids, glycine, lysine, and total amino acids were positively correlated with birth weight, whereas valine and threonine showed a negative correlation. Cranial volume at birth was significantly related to the level of glycine and alpha1 globulin. Other protein and amino acids which contributed to cranial volume included threonine, histidine, glutamic acid, and beta globulin, negatively correlated, whereas glutamine was positively correlated. Alpha1 globulin (negatively) and glycine (positively) made significant contributions to motor development. For mental development, alpha 1 globulin and histidine showed a negative correlation, whereas isoleucine was positively correlated. Findings indicated that the concentrations of certain maternal amino acids and proteins in the third trimester of pregnancy correlated significantly with fetal growth and development. This study suggests: (1) specific proteins or amino acids may be responsible for different developmental measures; (2) injudicious dietary restrictions in pregnancy should be avoided; (3) the determination of alpha1 globulin and a few amino acids such as glycine, lysine, and histidine in late pregancy may be used as predictors of fetal growth and development.


Antinuclear antibodies in chronic psychotic patients treated with chlorpromazine. The authors found a significantly higher incidence of antinuclear antibodies (ANA) and positive lupus erythematosus cell tests in chronic psychotic patients who received 400 mg a day or more of chlorpromazine (CPZ) for at least 7 weeks than in those who had taken 50 to 300 mg a day for varying periods or those who had received no CPZ for at least 3 months. Despite the high incidence of ANA, there was no observed development of lupuslike syndrome. The authors suggest that CPZ in high doses may induce ANA in humans.


A more reliable gram staining technic for diagnosis of surgical infections. The purpose of this study was to identify pitfalls in the Gram staining technic that limit its diagnostic value. In our clinical experience, gram-positive organisms were often decolorized too easily. Factors have been identified that alter the susceptibility of gram-positive organisms to decolorization in the Gram staining technic. The age of the bacterial culture, the preparation of the smear, the fixation technic, and the mordant have an important influence on the ease with which gram-positive organisms are decolorized. On the basis of these studies, a more reliable and reproducible Gram staining technic has been developed for the diagnosis of surgical infections.


Severe histamine mediated reactions to intravenous drugs used in anaesthesia. Severe histamine mediated reactions to intravenous drugs used in anaesthesia may occur as a result of anaphylactic and anaphylactoid reactions. The incidence is rare, but appears to be increasing. The difficulties in diagnosing such reactions and in determining the drug responsible and how these difficulties have led to confusion in the literature are discussed. Six cases of severe histamine mediated reactions are presented and detailed analyses of the drugs in these and other reported reactions is made showing varied clinical patterns with different drugs. The prevention, treatment, and follow-up of severe histamine mediated reactions are considered.


[The budgerigar (Melopsittacus undulatus)--a suitable endocrinological model of the thyroid gland]. The budgerigar (Melopsittacus undulatus) was tested as an endocrinological model of the thyroid gland. For this purpose, 514 animals were applied in our investigation. Besides the elaboration of an animal-specific method for the registration of thyroid function, questions of iodine metabolism, especially the influence of several endogenous and exogenous factors, were elucidated. Taking into consideration the results received we examined the model with regard to its application in functional, morphological, pharmaceutical, and radiotherapeutical questions. The results show that, by means of the method used, the model answers the various questions unreservedly. Further advantage recommend the budgerigar as a model of the thyroid gland.


Palliation in malignant pleural effusion. Experience with thoracostomy and intrapleural chemotherapy in the management of malignant pleural effsion is presented. Of 19 patients not requiring further thoracentesis, five died within one month. The surviving 14 patients all responded, 11 completely and three partially, with good palliation, for periods of from one to 28 months. Indications for the procedure and technical problems are discussed, and the results evaluated. This would seem ot be the method of choice for recurring malignant pleural effusion.


Some aspects of immunohistoperoxidase reactions in the agarose bead model system. Agarose beads that serve as matrices for antigens can be useful tools, not only for studying the quantitative aspects of immunohistoenzyme reactions but also for detecting antibodies. The present study focuses on localization of the ultimate reaction product within the beads; this factor is important when quantitative aspects of immunohistoenzyme reactions are under investigation. Localization varies with respect to the concentration of CNBr used to activate the beads. In addition, a method is described that facilitates the use of indirect immunohistochemical bead procedures and reduces the amount of antigen (-coupled beads) needed. These advantages may be particularly helpful when these methods are used for serologic detection of specific antibodies on a large scale.


Diagnosis of Neisseria infections by defined immunofluorescence. Methodologic aspects and applications. Defined IF techniques have been used in our laboratories for the past 15 years for diagnosing meningococcal and gonococcal infections. Conjugates against these bacteria give desired specific staining (DSS) but also undersired specific staining (USS) due to cross-reacting antigens. USS is controlled by absorption. Nonspecific staining (NSS) of protein A-containing S. aureus caused by the reaction with the Fc part of the IgG molecule is blocked by adding RB 200-labeled antiS. aureus globulins to diluted FITC conjugates against gonococci or menigococci. Nss is also avoided by using FITC-labeled F(ab')2 fragments of IgG. By optimal labeling and separation of unlabeled FITC and heavily labeled molecules on Sephadex G-25 at low ionic strength and low pH, nonspecific reactions with tissues are reduced to a minimum. Defined IF can be utilized for diagnosing pathogenic Neisseria by examination of smears prepared from clinical specimens, as confirmative tests of oxidase-positive colonies, and for demonstrating these bacteria in tissue efflorescences and tissue biopsies. It is stressed that this technique should always be used in conjunction with conventional methods.


Rhodamine conjugates:specific and nonspecific binding properties in immunohistochemistry. Anionic-exchange fractions of IgG labeled with FITC, MRITC, RB200SC, or RBITC were tested on different substrates, and the resultant fluorescence was evaluated with the Ploem optical system. Conjugations with MRITC or RB200SC were found to afford the following advantages over FITC: immunofluorescence sensitivity was elevated six to seven times on a molar basis; high sensitivity could be combined with a wide specificity interval; there was negligible fading of emitted light; there was negligible tissue autofluorescence at the excitation wavelength (546 nm); because of the two latter points, repeated observations could be made on tissue sections stored for several years; and eosinophilic leukocytes that are prone to yield nonspecific staining could easily by identified by switching to ultraviolet-blue light excitation.


Immunology of DNA. III. Crithidia luciliae, a simple substrate for the determination of anti-dsDNA with the immunofluorescence technique. C. luciliae are hemoflagellates nonpathogenic for man and easy to culture. They have a giant mitochondrion, in which the mitochondrial DNA is concentrated in a single large network, the kinetoplast. When used as a substrate for the indirect immunofluorescence technique, studying sera from patients with SLE, we could demonstrate a very good correlation between this test and the Farr assay for the demonstration of antibodies to double-stranded DNA. Although the sensitivity of both techniques is on the same order of magnitude, the IF technique has the following advantages over the Farr assay. It is easy to perform in laboratories equipped for autoimmune serology. It possesses an intrinsic check on the immunoglobulin character of the DNA-binding activity. It allows one to determine the Ig classes and subclasses of antibodies to DNA. It permits study of complement fixation to antibodies without interference of Clq fixation to DNA or anticomplementarity of the serum. There is an absence of interference with antibodies to single-stranded DNA.


An approach to quantitation in rhodamine isothiocyanate labeling. A survey of commercially available TMRI and RBI products revealed that many are of unsatisfactory quality. Optimum dye/IgG ratios were obtained in conjugates with TMRI of about 65% purity by labeling them with reaction mixture ratios of 10 and 20 mug dye/mg IgG. Optimum labeling with RBI of comparable purity required reaction mixture ratios of at least 20 mug dye/mg IgG. Although specific storage stability studies of TMRI or RBI products were not performed, one TMRI product underwent appreciable degradation during a 2-year period of storage in a desiccator over Drierite. The importance of evaluating the protein labeling capacity of TMRI or RBI products before making immune conjugate preparations is evident.


Studies on defined immunofluorescence in clinical immunopathology. II. Relationship of F/P ratios pf conjugates and staining properties in indirect immunofluorescence. The effect of conjugate F/P ratios on titers of patient antibodies was evaluated with two model systems in IIF chessboard titrations, notably, antinuclear antibodies and the intercellular antibodies of pemphigus. The plateau titers of both antibodies and F/P conjugate ratios were analyzed and proved to have a high degree of linear relationship statistically. Single titrations of other tissue antibodies, including skin basement membrane antibodies of bullous pemphigoid, MTA, GPA, SMA, and ThA with the same group of conjugates yielded similar information. A quantitative relationship between the two parameters was established.


Microfluorometric evaluation of conjugate-specificity with the defined antigen substrate spheres (DASS) system. Six fluorescent antihuman Ig preparations were tested for their Ig class specificity by reacting them with highly purified IgG, IgM, IgA, and OVA coupled covalently to Sepharose beads. OVA was used as a measure for nonimmunologic binding. Bead fluorescence was determined by microfluorometry. The amounts of USS and NSS were expressed quantitatively. These data were compared with the performance of these particular conjugates in a biologic system, namely, monoclonal bone marrow cells. Five of the six conjugates satisfied the requirement of monospecific activity; one did not. At a dilution of 1 : 8, the five monospecific conjugates reacted between five and 50 times stronger with their appropriate antigens than with OVA-coupled beads. Cross reactivity with other Ig classes, after correction for OVA staining was maximally 6%. The conjugate that was nonspecific in the  bone marrow system gave very high cross reactivity with the Ig-coupled beads. A good  correlation was found between OVA bead staining and nonimmunologic binding of conjugates in bone marrow slides. In this respect, conjugates prepared from antibody preparations isolated by solid immunoadsorbents proved to be superior to globulin or whole IgG fractions. Ig coupled to Sepharose beads seems to represent a very promising substrate for conjugate specificity testing.


Studies of the linkage relationship of beta-2-microglobulin in man-mouse somatic cell hybrids. Beta-2-microglobulin (beta2mu) production has been studied in 33 primary man-mouse hybrid clones and in 26 secondary man-mouse hybrid clones. These clones have also been examined for the presence of 15 human enzyme phenotypes. Karyotypic analyses have been carried out on clones. From these studies the following conclusions can be drawn: (1) Gene(s) determining human beta2mu production in humans are apparently syntenic with the MPI gene on chromosome 15. (2) Long-term fibroblast lines may be of limited use in mapping studies as chromosomal rearrangements frequently occur in these lines. (3) The gene(s) determining beta2mu production in humans segregate independently of chromosome 6. If the assignment of the genes determining HL-A alloantigens to chromosome 6 is correct, our results imply that beta2mu and HL-A alloantigens are determined by genes carried on different chromosomes, despite the fact that beta2mu forms an integral part of the HL-A molecule.


Properties of bispecific rosette-forming cells. I.--Presence of T-lymphocyte markers. Three T-cell markers have been used to determine the origin of bispecific rosette forming cells (RFC) observed after stimulation by two species of heterologous red cells. The inhibition by anti-theta serum (AOS), azathioprine (AZ) and anti-lymphocyte serum (ALS) of the majority of bispecific RFC indicates their T-cell origin. The study of their sensitivity to these inhibitors suggested that about half of the bispecific RFC belong to immature T1 cell subset and the other half to more mature T2 cells. Conversely, monospecific RFC include both B-celle RFC have characteristics different from monospecific RFC argues against the reaction of double RFC against a common antigenic determinant of the two erythrocyte types.


Properties of bispecific rosette-forming cells. II. --Rosette formation by "educated" T-lymphocytes. Normal thymus cells transferred into thymectomized and lethally irradiated syngeneic mice responded to stimulation with sheep erythrocytes by rosette formation with these erythrocytes. This response reached a peak on the 7th day and was not associated with any production of circulating antibodies. Rosettes produced by "educated" T-cells were inhibited by anti-theta serum as well as by anti-mouse Fab serum. Simultaneous stimulation of transferred thymocytes with sheep and pigeon erythrocytes provoked the appearance of a minority of cells simultaneously binding both types of erythrocytes. Depletion of B cells contaminating normal thymocyte populatins after passage through anti-mouse Ig coated columns before their transfer in thymectomized and irradiated recipients did not prevent the appearance of simple and double RFC. Moreover, when normal thymocytes or T-cells "educated" by allogeneic stimulation were incubated at 4 degrees C with anti-SRBC and anti-PRBC mouse sera a subsequent incubation at 37 degress of resulted in the dissociation of most passive rosettes formed at 4 degrees. Conversely similar incubation at 37 degrees of rosettes formed by actively immunized cells resulted in capping of about 50% simple and double rosettes. This redistribution of membrane receptors is proposed as a routine test for distinguishing active from passive rosettes.


Developments in laboratory diagnosis of chromosomal abnormalities. The development of chromosome banding techniques has made possible the detailed analysis of human chromosomes. A vast body of knowledge has accumulated during the last few years as a result of their application in biology and medicine.


Two IgG1 (IgF) subclasses in mice: presence in normal serum and representation within lymphocyte and plasma cell populations. Monospecific anti-gamma1 sera were prepared in rabbits against each of two IgG1 mouse myeloma globulins isolated from myeloma A2 and MOPC21. Both these sera react with IgG1 of normal mouse serum by immunoelectrophoresis and cannot be distinguished any further. The antibodies contained in the sera were cross-isolated on immunoadsorbents containing either the A2 or MOPC21 antigenic material and tested by immunoelectrophoresis against normal mouse serum. An interesting spur was formed between these two antibodies. The two isotypes thus revealed in normal serum are tentatively designated IgG1a (globulin A2) and Ig G1b (globulin MOPC21). With live peripheral lymph node cells almost all lymphocytes which stain with antikappa antibodies (B lymphocytes) also stain with the specific anti-gamma1a antibodies, but only 5-10% stain with the specific anti-gamma1b antibodies. On fixed spleen cells (C57B16 mice) the anti-gamma1a antibodies stain almost identical percentages of plasma cells as anti-kappa antibodies: 1.8% are gamma1 and 2.2% are kappa. Anti-gamma1b antibodies stain only 5% of kappa-positive plasma cells. The % values estimated on both the normal lymphocyte and plasma cell populations were not significantly affected by inhibtion experiments of the specific anti-gamma1 antibodies performed with isolated myeloma globulins representative of several Ig classes.


[Electron microscopic study of the localization of penicillin acylase in E. coli cells]. The paper describes the studies on definition of penicillinacylase localizations in the cells of E. coli with the help of ferritin labeled immune sera and electron microscopy. Both the intact cells and the cells treated with the substances affecting the cell wall intactness were used. The study showed relation between penicillinacylase and the surface structures of the cell, i.e. the cell wall and the periplasmic areas. It was found that penicillinacylase got into the environmental medium with the splitted cell fragments which corresponded to the general mechanism of excretion of large high molecular compounds by gramnegative organisms.


Concentration of Rous sarcoma virus from tissue culture fluids with polyethylene glycol. Concentration of Rous sarcoma virus from tissue culture fluids with polyethylene glycol, with and without NaCl or dextran sulfate, resulted in significant and highly variable losses caused by entrapment of virus particles in proteinaceous debris. Treatment of concentrated preparations with Pronase greatly enhanced the recovery of virions. Maximum recovery of virus particles was obtained by the addition of 8% polyethylene glycol and 0.4 M NaCl to tissue culture fluids, followed by Pronase treatment of the concentrated virus preparations.


Automated single-slide staining device.

Automated single-slide staining device. An automated single-slide staining device is described. Smears gram stained by the automatic device were equal in quality to the manual method.


The Blalock-Hanlon procedure. A new look at an old operation. The Blalock-Hanlon procedure has been carried out in 48 neonates with 10 early deaths (21%). This compares favorably with the reported early mortality for balloon atrial septostomy. Most striking have been the length of palliation, freedom from complications, and low incidence of late death among these patients. In contrast, reported series of balloon septostomies have shown a high incidence of strokes, repeat hospitalization, inadequate palliation, and late deaths. Since palliation is reliable following a Blalock-Hanlon procedure, we do not favor early correction with hypothermic arrest but recommend it at 12 to 18 months of age, when conventional perfusion can be easily utilized.


The influence of non-cytotoxic concentrations of the herbicide 2,4-dichlorophenoxyacetic acid on the DNA synthesis in cultured vertebrate cells. The influence of the herbicide 2,4-Dichlorophenoxyacetic acid on the growth of cultured chicken muscle cells was studied by observing morphological changes and by cytophotometric measurement of the DNA content of individual cells. DNA-synthesis curves were derived from the distribution of DNA values. No marked effects were observed at a concentration of 60 mug per liter. In contrast, doses of 600 mug per liter caused a loss in cell differentiation consisting of a partial lack of polar orientation, diminished formation of fibers, and an increase of the N/C ratio. The nuclear matrix of desoxyribonucleoprotein (DNP) changed to a more heteropycnotic structure. Isolated cell necroses were found after 44 hrs of exposure. When compared to the controls, the relative DNA synthesis time was found to be prolonged 1.5-fold. The number of cells in the G2 phase decreased during 44 hrs of exposure. It is concluded from these results that the dosedependent growth inhibition of 2,4-D on vertebrate cells in culture takes effect during DNA synthesis. The morphological changes of the nuclear structure suggests that 2,4-D directly affects the template activity of the DNP matrix.


[Quantitative fluorometric determination of gamma globulin and fibrin in the aortic wall of rabbits with experimental atherosclerosis]. By means of the fluometric method in 76 rabbits at definite stages of the development of experimental atherosclerosis in the aorta an increased amount of gamma-globulin and fibrin was established. An increase in the level of gamma-globulin was accompanied by fixation of beta1C-fraction of complement, which is characteristic of deposit of an immune complex. The most pronounced concentration of gamma-globulin was noted in areas of lysis and fragmentation of elastic fibres.


[Total transposition of the great vessels: Surgical treatment]. One hundred nine cases of complete transposition of great vessels alone or associated to other abnormalities are presented. In 91 cases a palliative treatment is given and in 18 cases there is a total correction done, depending upon the technique. Surgical indication and the technique used in every case is commented, making special reference to transpositions with ductus arteriosus or associated to coarctation of the aorta. In the 18 cases where we have done a total correction, we comment the surgical procedure of one of them who had a subpulmonary stenosis of the fibromuscular type, which we resolved through left ventriculotomy done on the apex of the ventricle and resecting the stenosis with an electroscalpel, which is a technique we have devised.


Vascular changes in the surrounding of osteoid osteomas. Authors observed in 11 instances of osteoid osteoma out of 20 revised cases the characteristic features of arteriolosclerosis in the small arteries supplying the nidus. With these, earlier hypothesis of Jaffe about an increased intravascular pressure in the nidus and supplying vessels in supported.


Studies on the role of the areA gene in the regulation of nitrogen catabolism in Aspergillus nidulans. Mutants of Apergillus nidulans with lesions in a gene, areA (formerly called amdT), have been isolated by a variety of different selection methods. The areA mutants show a range of pleiotropic growth responses to a number of compounds as sole nitrogen sources, but are normal in utilization of carbon sources. The levels of two amidase enzymes as well as urease have been investigated in the mutants and have been shown to be affected by this gene. Most of the areA mutants have much lower amidase-specific activities when grown in ammonium-containing medium, compared with mycelium incubated in medium lacking a nitrogen source. Some of the areA mutants do not show derepression of urease upon relief of ammonium repression. The dominance relationships of areA alleles have been investigated in heterozygous diploids, and these studies lend support to the proposal that areA codes for a positively acting regulatory product. One of the new areA alleles is partially dominant to areA+ and areA102. This may be a result of negative complementation or indicate that areA has an additional negative regulatory function. Investigation of various amdR; areA double mutants has led to the conclusion that amdR and areA participate in independent regulatory circuits in the control of acetamide utilization. Studies on an amdRc; areA double mutant indicate that areA is involved in derepression of acetamidase upon relief of ammonium repression.


The effects of allopurinol and clofibrate on the elimination of coumarin anticoagulants in man. The effects of chronic treatment with allopurinol and clofibrate on the elimination from plasma of warfarin and dicoumarol were examined in man. In addition, the binding of warfarin to human serum proteins was measured with and without clofibrate and its metabolite chlorophenoxyisobutyric acid (CPIB), both present in therapeutic concentrations. Treatment with allopurinol and clofibrate did not alter the rate of elimination of warfarin from plasma. In addition, clofibrate and CPIB caused no significant displacement of warfarin from serum proteins. This evidence supports the conclusion that the clinically significant potentiation of warfarin activity by clofibrate in man is due to an interaction at the receptor site. In contrast, treatment with allopurinol resulted in significant prolongation of the plasma half-life of dicoumarol in one of three subjects. These data are consistent with inhibition of the metabolism of dicoumarol by allopurinol in some individuals.


Comparative ability of hybrid and homozygous resistant larvae of Lucilia cuprina to cause strikes under experimental conditions. The intervals between the jetting of sheep with dieldrin or BHC, and the inducement of flystrike by implants of hybrid and homozygous BHC/dieldrin resistant larvae of Lucilia cuprina, were compared. Flystrikes soon established with homozygous resistant larvae, but considerable protection was afforded against hybrid larvae, the interval being longer with BHC than with dieldrin. Dieldrin was shown to have a residual effect on rr larvae in a mixed population 9 weeks after jetting. It is suggested that wide adoption of mulesing and mid-season's crutching would favour reversion to susceptibility in the blowfly and thus extend the field life of insecticides to which resistance has emerged.


Some staining characteristics of Mott's or morular cells of nagana. The spherules of morular (Mott's) cells are not metachromatic with toluidin blue at pH 8 and the majority are stained yellow with PTAH. This distinguishes them from globule leucocytes. They differ from Russell bodies of plasma cells which are Gram-positive.


Use of phast contrast microscopy of semithin epoxy sections in histochemistry. A number of histochemical staining methods can be applied to semithin (1-2 mum) Epon sections either unmodified or after slight modification of the original method. While only the histochemically stained specific component is visible in bright field illumination, phase contrast microscopical study of the same field of view shows also the histological and cytological details of the specimen and thus allows exact localization of the specific component in the tissue. Applications will be found in several areas in histochemistry. Practical examples are given for demonstration of periodic acid-SCHIFF-reactive mucosubstances, exogenous perioxidase, and acid phosphatase activity.


Further studies on the specificity of the N- and C-terminal antigenic determinant of hen egg-white lysozyme. The specificity of the N- and C-terminal antigenic determinant (P17: sequence Lys1-Cys6-Asn27, Trp123-Cys127-Leu129) of hen egg-white lysozyme (HL) was studied in more detail. In a Scatchard plot of the binding of 14C-acetyl HL with guinea pig purified anti-P17 antibody experimental values bent sharply near r=1. This suggests the presence of two antibody populations with different affinities for HL or possible steric hindrance in the binding of a second HL molecule to the second binding site of the antibody molecule. The antigenic activities of various peptides were tested by measuring their inhibition of the binding of 14C-acetyl-P17 with the anti-P17 antibody. Only P17 and P17t (sequence Lys1-Cys6-Homoser12, Trp123-Cys127-Leu129) were inhibitory, with KI values of 2.0 times 10(4) and 8.1 times 10(3), respectively. These results indicate that the direct binding site of P17 to anti-P17 antibody may be located in the terminal portion of P17 (sequence Lys1-Cys6-Homoser12, Trp123-Cys127-Leu129) while the rest of P17 may be important in maintaining the conformation of this determinant. The single disulphide blood involved in this determinant is essential for manifestation of immunological activity.


Comparative studies on antibody formation against hen egg white lysozyme and its split fragment. A peptide fragment Fr. 17 (Lys1-Cys-Asn27 Leu129-Cys-Ala122) of hen egg white lysozyme (HL) was previously found to retain at least one antigenic determinant region of the native protein. In this work a highly purified preparation of Fr. 17, contaminated with less than 0.01% HL and less than 1% of other fragments was found to be strongly immunogenic to rabbits. The kinetic patterns of antibody formation against Fr.17, assayed by passive hemagglutination (PHA), were quite different from those of antibody formation against HL. The specificity of the antibody elicited to Fr. 17 was mainly directed to the Fr. 9-10-a region (Ala11-Asn27) while that of the antibody elicited to the Fr. 17 region in native HL was directed to the Fr. 15-b region (Lys1-Cys-Ala10 Leu129-Cys-Trp123). It is concluded that in the process of antibody formation, the recognition of the Fr. 17 region in native HL is different from that of fragment Fr. 17.


Discrimination of antigenic site and thiol-inhibitor-sensitive site of hexokinase isoenzymes. Rabbit antiserum was prepared against hexokinase isoenzyme type I which was purified from rat brain mitochondria. The antiserum inhibited the activity of the mitochondrial hexokinase type I as well as that of the cytosolic type I enzyme prepared from rat brain, kidney and spleen. It did not, however, inhibit the activity of type II hexokinase from muscle and spleen or that of the type III enzyme from spleen. The results suggest that all hexokinase type I isoenzymes may have a common antigenic site irrespective of their sources, though their responses to a thiol inhibitor are different.


In-vitro inhibition of RNA-instructed DNA polymerase. Rifamycins and distamycins were assayed in vitro for their effects on the activity of a reverse transcriptase derived from the plasma of a patient with breast cancer. The inhibitory effect observed was compared with that of a human placental substance with a molecular weight of approximately 10 000. The latter substance is apparently also capable of affecting reverse transcriptase activity in vitro.


The effect of specific brain phospholipid on the rat's allergic encephalomyelitis. A new preparation which contains specific brain phospholipids (Gricertine) acts as a histamine liberator in CNS and protects rats from allergic encephalomyelitis. It is supposed that the protective effect of Gricertine on the allergic encephalomyelitis is based on its histamine-liberator properties which can occur during the first days of therapy: this would explain the large decrease of the brain histamine after 3 weeks of treatment. Gricertine is also able to remove clinical symptoms of allergic encephalomyelitis in rats. It is presumed that Gricertine may also be acting independently of that mechanism, in the way that it replaces phospholipids that disappeared in the cases where demyelinisation had taken place.


[Sensitivity tests of malignant tumours against cytostatic agents in vitro and in vivo/studies on the mouse sarcoma 180 (author's transl)]. The effects of ten different substances on the mouse sarcoma 180 have been compared using in vivo and in vitro test systems. The size of the tumours was taken as a measure of the success of the therapy in animal experiments. The in vitro effects were estimated by measuring the incorporation of radioactively labelled uridine in tumour cell suspensions from solid tumours, in ascites tumours and in tissue culture. Similar results were obtained using all three in vitro test systems. Four substances (daunomycin, fluoruracil, actinomycin D, adriamycin) exhibited activity both in vivo and in vitro, whereas five substances (cytosinarabinoside, methotrexate, ibenzmethyzin, triaziquone, bleomycin) showed no activity in any of the test systems used. With podophyllic acid ethyl hydrazide, however, no correlation between in vivo and in vitro effects was observed. Studies on the influence of the cytostatic agents on the rate of transport of uridine into the cells showed that podophyllic acid ethyl hydrazide strongly reduced the permeability of the cells to radioactive uridine.


Metabolism of a prednisolone compound in human under normal and pathological conditions. A method has been presented for the quantitative determination of 21-deoxy-N-(N'-methyl-piperazinyl)-prednisolone (Depersolone), a water soluble corticosteroid, and one of its metabolites, prednisolone, in human urine. Following administration of 30 or 90 mg Depersolone i.v., the dynamic appearance of this hormone and the prednisolone metabolite, their biological half-lives and the effect of the synthetic corticosteroid on the excretion of grouped and individual 17-ketosteroids were determined. Results of experiments revealed a dose dependent excretion of the unchanged hormone and a biological half-life 12 to 23 h in control subjects. Prednisolone as a metabolite seems to play a minor role in Depersolone action. The excretion rate of Depersolone was greater and its half-life less than normal in hepatopathies. The dynamic appearance of the hormone was found slow in the urine of patients with prolonged corticosteroid therapy.


Cytological and functional criteria for the classification of malignant lymphomata. The subtle morphology and functional properties of cells are the best parameters to use for their definition. This is also true for the corresponding tumours, especially malignant lymphomata. In studies of 106 cases of malignant lymphoma we therefore applied as morphological methods haematological staining (Giemsa in sections and imprints) and electron microscopic analysis. As functional criteria we used the nonspecific esterase reaction to define tumours of histiocytes and an estimation of the immunoglobulin content of tissue extracts and single cells to define tumours of B lymphocytes and their derivatives. By combining all of these methods it was possible to propose a new classification. Whereas not one histiocytic malignant lymphoma ("reticulosarcoma") was found in the series, at least most of the malignant lymphomata investigated seemed to be derived from the B lymphocyte system. The following types are distinguished: (1) Chronic lymphocytic leukaemia; (2) diffuse germinocytoma (malignant lymphoma, lymphocytic, intermediate); (3) germinoblastoma (follicular, follicular and diffuse, diffuse; sclerotic, nonsclerotic) which can show a transition into germinoblastic sarcoma; (4) immunoblastic sarcoma of the B cell type (previously called reticulo-sarcoma); (5) lymphoplasmocytoid immunocytoma, which may be associated with Waldenstrom's macroglobulinaemia and can show a mixed cellularity; (6) lymphoblastic (paraleukoblastic) sarcoma and leukaemia, which are, at least in most cases, probably neoplasias of germinoblasts. All of these lymphomata can produce immunoglobulins. Sixty-seven cases showed an Ig increase in the tumour. This was mostly IgM, but sometimes IgG, IgA, IgD and/or IgE. A morphological equivalent of even abnormal Ig secretion is the globular positive (diastase resistant) PAS reaction in lymphoid cells (not in the histiocytes) in paraffin sections, which we found in 43 cases. Only 19 of the 63 cases with an IgM increase  in the tumour showed an increase of IgM in the serum. Waldenström's macroglobulinaemia is a facultative symptom of morphologically different malignant lymphomata and should therefore be considered only as a clinical syndrome and not as a nosological entity.


Combination chemotherapy and radiotherapy in non-Hodgkin's lymphomata. The results obtained with intensive chemotherapy and intensive chemotherapy plus radiotherapy in non-Hodgkin's lymphomata are reported. A quintuple drug regimen (mechloretamine, adriamycin, bleomycin, vincristine and prednisone) in histiocytic lymphomata (Stage III and IV) yielded complete remissions in 53% and complete plus partial remissions in 77%. These figures were 44% and 64% respectively in lymphocytic lymphoma. In Stage III complete responders after combination chemotherapy were subsequently irradiated (involved field irradiation). The median duration of complete remission after completion of radiotherapy was 9-5 months in histiocytic and 12-0 months in lymphocytic lymphomata. At 2 years actuarial survival in Stage III and IV was better in patients with the lymphocytic type and with nodular pattern than with histiocytic and diffuse patterns. A more recent trial compares, in Stage IV patients, cyclophosphamide, vincristine and prednisone (CVP) versus adriamycin, bleomycin and prednisone (ABP). Although the number of evaluable patients is still limited, there appears to be no difference in the response rate between CVP and ABP. In Stages I and II, 6 cycles of CVP were given as adjuvant treatment after radiotherapy. At the present moment, there is no statistical difference in the relapse rate between the group of patients treated with radiotherapy alone and that with radiotherapy plus CVP.


Non-Hodgkin's malignant lymphomata in adults: chemo-radiotherapy in stages III and IV. Patients presenting with Stage III or IV non-Hodgkin's malignant lymphoma were given chemotherapy; about 20% complete remission was obtained for both stages. The addition of radiotherapy increased the incidence to 70% in Stage III patients. The duration of first complete remission was longer for Stage III (25% of the patients are still in first remission at 7 years) than for Stage IV (0%). The survival was longer for nodular lymphosarcoma patients (25% are alive at 7 years for Stages III and IV) than for diffuse lymphosarcomata and reticulosarcomata (10%). Among the new drugs, VM 26 is able to produce a good frequency of remission in patients in relapse.


Light and electron microscopic differentiation of amyloid and colloid or hyaline bodies. Hyaline or colloid bodies (Civatte bodies) and amyloid are sometimes confused because of their similar appearance under the light microscope. A differentiation can be made by histochemical means (DMAB-Nitrite method) and by fluorescence or polarization microscopy. At the ultrastructural level hyaline or colloid bodies as well as amyloid exhibit a fibrillar structure, but can be differentiated by the diameter and arrangement of fibrils and the localization of the aggregations. A clearcut differentiation between hyaline or colloid bodies and amyloid seems of great importance in any dicussion of the epidermal origin of amyloid and the classification of fibrillar aggregations associated with skin tumours.


Maternal plasma alphafetoprotein levels in the second half of normal pregnancy: relationship to fetal weight, and maternal age and parity. A semi-automated radioimmunoassay for plasma alphafetoprotein has been developed, suitable for routine clinical use. Several hundred samples can be assayed in a working week, with results available within 24 hours. The use of extensive quality controls ensures that good precision is maintained both within and between assays. The range of plasma concentrations of alphafetoprotein in the second half of pregnancy has been established in 100 normal subjects. The levels rise progressively to reach a peak at 32 weeks, and thereafter fall until term. No relationship between circulating alphafetoprotein levels and birth weight was observed.


A semi-automated serum alphafetoprotein radioimmunoassay for prenatal spina bifida screening. A simple, reliable, semi-automated and inexpensive radioimmunoassay for serum alphafetoprotein (AFP) is described which fulfils the requirements for a screening test for open neural tube defects. The normal range for serum AFP throughout pregnancy has been determined in 1534 samples. In a retrospective study of 15 samples of maternal serum taken from nine pregnant women carrying a fetus affected with a neural tube defect, the serum AFP level was found to be above the 95th centile in four of the five cases of open defect sampled between 15 and 20 weeeks gestation. Preliminary results of a prospective study are reported in which two cases of nencephaly were detected among 671 unselected pregnancies sampled between 15 and 20 weeks gestation.


Detection of structural differences between nuclear and mitochondrial glutamate dehydrogenases by the use of immunoadsorbents. Structural differences between crystalline mitochondrial and nuclear glutamate dehydrogenases from ox liver have been detected by immunological techniques. Antisera prepared against each enzyme precipitate both glutamate dehydrogenases; upon immunodiffusion, the antiserum against the nuclear enzyme gives a line of incomplete identity with the two antigens, whereas the antiserum against the mitochondrial enzyme gives a line of complete identity. Fractionation of the antibodies contained in each antiserum by means of an immunoadsorbent, to which the nuclear or the mitochondrial enzyme has been covalently linked, shows that nuclear glutamate dehydrogenase (GDH) contains specific antigenic determinants as well as determinants common to the mitochondrial enzyme, whereas the latter appears to have no antigenic portions which are not present in the nuclear antigen, in accord with the results of immunodiffusion. The antibodies against determinants common to both enzymes precipitate and inhibit them, whereas the specific anti-nuclear GDH antibodies precipitate but do not inhibit the nuclear antigen.


Effects of proteins on thermotropic phase transitions of phospholipid membranes. A variety of proteins have been studied for their ability to interact and alter the thermotropic properties of phospholipid bilayer membranes as detected by differential scanning calorimeter. The proteins studied included: basic myelin protein (A1 protein), cytochrome c, major apoprotein of myelin proteolipid (N-2 apoprotein), gramicidin A, polylysine, ribonuclease and hemoglobin. The lipids used for the interactions were dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylglycerol. The interactions were grouped in three catagories each having very different effects on the phospholipid phase transition from solid to liquid crystalline. The calorimetric studies were also correlated with data from vesicle permeability and monolayer expansion. Ribonuclease and polylysine which exemplify group 1 interactions, show strong dependence on electrostatic binding. Their effects on lipid bilayers include an increase in the enthalpy of transition (deltaH) accompanied by either an increase or no change in the temperature of transition (Tc). In addition, they show minimal effects on vesicle permeability and monolayer expansion. It was concluded that these interactions represent simple surface binding of the protein on the lipid bilayer without penetration into the hydrocarbon region. Cytochrome c and A1 protein, which exemplify group 2 interactions, also show a strong dependence on the presence of net negative charges on the lipid bilayers for their binding. In contrast to the first group, however, they induce a drastic decrease in both Tc and deltaH of the lipid phase transition. Furthermore, they induce a large increase in the permeability of vesicles and a substantial expansion in area of closely packed monolayers at the air-water interface. It was concluded that group 2 interactions represent surface binding followed by partial penetration and/or deformation of the bilayer. Group 3 interactions, shown by proteolipid apoprotein and gramicidin A, were primarily non-polar in character, not requiring electrostatic charges and not inhibited by salt and pH changes. They had no appreciable effect on the Tc but did induce a linear decrease in the magnitude of the deltaH, proportional to the percentage of protein by weight. Membranes containing 50% proteolipid protein still exhibited a thermotropic transition with a deltaH one half that of the pure lipid, and only a small diminution of the size of the cooperative unit. It was concluded that in this case the protein was embedded within the bilayer, associating with a limited number of molecules via non-polar interactions, while the rest of the bilayer was largely unperturbed.