Surface mapping of mouse thymocytes.

Surface mapping of mouse thymocytes. The blocking method used previously for determining the relative positions of different components of the cell surface was modified by first fixing the cells with paraformaldehyde. This technique was applied to the H-2K (K), H-2D (D), TL, Lyt-1, and Lyt-2 surface components of mouse thymocytes, and the results were compared in parallel with data obtained with the original technique with unfixed cells. Previous mapping data with unfixed cells, indicating the positions of these molecules relative to one another, were confirmed with paraformaldehyde-fixed cells, with one exception. On unfixed cells, D and TL appeared sufficiently adjacent to produce mutual interference in the attachment of anti-D and anti-TL antibodies. With paraformaldehyde-fixed cells this was not so, D and TL appearing sufficiently separated from one another to obviate interference in the attachment of anti-D and anti-TL antibodies. The previously reported close association of K with Lyt-1 and of D with Lyt-2 were demonstrable equally with unfixed and paraformaldehyde-fixed thymocytes. It is suggested that activation of D sites, and alternatively of TL sites, by antibody in the present experiments brings these two molecules into apposition and that this movement may exemplify a mechanism concerned in immunological recognition and response.


Cell-mediated lysis of lipid vesicles containing eye muscle protein: implications regarding pathogenesis of Graves ophthalmopathy. We prepared artificial vesicles that are lysed upon cell-mediated immunological attack by human lymphocytes. These vesicles are made from a mixture of dimyristoyl lecithin, dipalmitoyl lecithin, and cholesterol, have eye muscle membrane protein (EMP) inserted into the bilayer wall, and contain intravesicular (99m)Tc marker. Injury to the vesicular membrane was assessed by measurement of (99m)Tc release. Thyroglobulin (Tg) and Tg-anti-Tg complex (TgA) bind to EMP-vesicles to an extent equal to or greater than to native eye muscle membranes in vitro; this binding requires the presence of normal human IgG. The role of Tg, TgA, IgG, and peripheral blood lymphocytes in altering membrane permeability was analyzed. Incubation of vesicles for up to 3 hr alone, with added IgG alone, or with further addition of Tg or TgA did not result in (99m)Tc release. Addition of lymphocytes from normal donors to the above four preparations showed release in the presence of TgA. Lymphocytes from each of eight patients with Graves ophthalmopathy caused release not only in the presence of TgA, but also in the presence of Tg. Separation of a patient's lymphocytes into high- and low-affinity rosette-formers (T and K cells, respectively) showed that cell-mediated vesicle lysis in the presence of TgA was greater with K cells than with T cells, while vesicle lysis in the presence of Tg was greater with T cells than with K cells. Vesicles made with inserted Tg but lacking EMP were not lysed by such T cells. Lymphocytes failed to induce permeability changes in vesicles containing other inserted proteins obtained from human nonextraocular muscle, liver, spleen, or adrenal, even if Tg or TgA were present. The results support the concept that muscle cell damage in Graves ophthalmopathy is immunological, cell-mediated, and of two types: (i) K lymphocytes reacting to immune complex, TgA, on the eye muscle cell surface (i.e., antibody-dependent cytotoxicity) and (ii) sensitized T lymphocytes reacting to Tg on the eye muscle cell surface. An antigenic role for EMP is possible, but has not been unequivocally proven.


Immunological tolerance: high-dose antigen-induced suppressor cells from tolerant animals inactivate antigen-presenting macrophages. Studies were carried out to characterize the target cell for the activity of suppressor cells induced in highzone tolerance to deaggregated human gamma globulin (HGG). We applied an in vitro system for the initiation of an immune response, consisting of culturing spleen lymphocytes on HGG-fed macrophages, in which initiator T cells are generated. These cells, when injected into the foot pads of syngeneic mice, recruit specific anti-HGG effector T lymphocytes. We found that HGG-fed macrophages were incapable of signaling spleen cells from HGG-tolerant animals to generate initiator cells. Spleen cells from tolerant animals, when mixed with spleen cells from normal donors, inhibited the capacity of the normal population to give rise to initiator cells after culture on HGG-fed macrophages. Thus, suppressor T cells, which inhibit education of T cells by antigen-fed macrophages, exist in the tolerant spleen. Spleen cells from HGG-tolerant animals, when seeded on macrophages fed simultaneously with HGG and keyhole limpet hemocyanin (KLH), also prevented the macrophages from signaling an anti-KLH response. Spleen cells from HGG-tolerant animals from which the suppressor cells were depleted by "affinity chromatography" on histamine columns, when seeded on macrophages fed with HGG and KLH, generated initiators to both antigens. It appears, therefore, that suppressor cells act at the level of antigen-presenting macrophages, affecting macrophages fed with the tolerogen, and therefore affecting also the immunogenic effect of other antigens presented by the same macrophages. By testing the mode of action of suppressor cells on the tolerogen-fed macrophage, we found that the suppressors manifest a cytotoxic effect on such macrophages. We propose that the suppressor cell is, in fact, an anti modified-self killer, acting on macrophages possessing surface self-antigens "modified" by the tolerogen. The similarity in cell-surface markers between suppressors and anti modified-self killers supports this concept.


Fine specificity of antibodies to poly(Glu60Ala30Tyr10) produced by hybrid cell lines. The polyethylene glycol-mediated cell fusion technique has been used to analyze the diversity of the antibody response to the terpolymer poly(Glu60Ala30Tyr10)(GAT). Nine stable clones (all producing IgM K anti-GAT antibodies) were isolated from a fusion between P3-X63-Ag8 myeloma cells and spleen cells from a DBA/2 mouse sensitized to GAT 5 days earlier. Seven other clones (producing IgG K anti-GAT antibodies) were derived from another fusion between NSI myeloma cells and spleen cells of (C57BL/6 X DBA/2)F1 hybrid mice hyperimmunized with GAT. These 16 anti-GAT antibodies were grouped according to their pattern of reactivity with GAT and the two related polymers of poly(Glu60Ala40) (GA) and poly(GLU50Tyr50) (GT). Two monoclonal anti-GAT antibodies (IgM F9-102.2 and IgG F17-148.3) demonstrated crossreactivity with GA but failed to crossreact with GT determinants. In contrast, the remaining 14 hybridoma antibodies demonstrated preferential reactivity with GAT but also exhibited crossreactive binding to GT and in some cases GA. There was a correlation between the fine specificity pattern and the presence of a common anti-GAT idiotype on these antibodies. Thus, the hybridoma anti-GAT antibodies which reacted with GT shared crossreactive idiotypic determinants (CGAT) present in mouse anti-GAT immune sera. In contrast, the monoclonal F9-102.2 and F17-148.3 antibodies that failed to bind to GT lacked the major CGAT idiotypic determinants.


Human breast carcinoma antigen is immunologically related to the polypeptide of the group-specific glycoprotein of mouse mammary tumor virus. We have shown [Mesa-Tejada, R., Keydar, I., Ramanarayanan, M., Ohno, T., Fenoglio, C. & Spiegelman, S. (1978) Proc. Natl. Acad. Sci. USA 75, 1529--1533] that an antigen immunologically related to gp52, a 52,000-dalton glycoprotein of the mouse mammary tumor virus, can be identified in sections of human breast cancer by means of an indirect immunoperoxidase technique. The specificity of the reaction was established by absorption experiments which revealed that only purified gp52, or material containing it, served to eliminate the IgG molecules responsible for the immunohistochemical reaction in the human breast tumors. We show here that the cross-reactivity between the human and murine tumor antigens is due to the polypeptide rather than the polysaccharide components of gp.52. Sugar-free gp52 prepared by deglycosylation with a mixture of glycosidases was as fully effective as the intact gp52 in removing from anti-MMTV the IgG responsible for the reaction with the human tumor antigen. In contrast, the isolated polysaccharide of gp52 was unable to exert blocking activity.


Ultrasonic assessment of the fetal spine before 20 weeks' gestation. The ultrasonic appearance of the normal and abnormal fetal spine before 20 weeks' gestation was reviewed in 121 pregnances. A model was constructed to establish the mechanism of production in the various configurations seen. The relative roles of ultrasonography, alpha fetoprotein assay, and fetoscopy in the diagnosis of spina bifida are discussed.


The extra-articular deposition of fibrinogen antigenic material in patients with rheumatoid arthritis. Experimental suction blisters were elicited on extensor and flexor sites or forearm skin of patients both with rheumatoid arthritis and without rheumatoid diseases. The blister fluid obtained was investigated, together with rheumatoid bursa fluid and rheumatoid noduli material by crossed and quantitative immunoelectrophoresis for fibrinogen antigenic material, plasminogen and inhibitors of fibrinolysis. In rheumatoid noduli and rheumatoid bursa fluid, fibrinogen degradation products identical with those previously described in rheumatoid synovial fluid and in bullous dermatosis were found (3, 14). On the other hand, no fibrinogen antigenic material was found in the experimental suction blisters. These results indicate that the fibrinogen degradation products in rheumatoid inflammatory exudate are probably not specific for a rheumatoid process, but rather part of a general mechanism in inflammation.


Acute myelomonocytic leukemia: coexistent cytochemical markers for monocytes and granulocytes in the leukemic cells. A 22-year-old man presented with mediastinal lymphoma and lymphoblastic leukemia. The leukemic cells in his blood and marrow did not stain positively with Sudan black B, peroxidase, chloroacetate esterase, or nonspecific esterase, and were considered as either lymphoblasts or stem cells. One year after the initial presentation, acute myelomonocytic leukemia developed and he died. The leukemic cells possessed the morphologic and cytochemical characteristics of both monocytes and granulocytes. This case illustrates the close ontogenetic relationship between the monocytes and the granulocytes. It also demonstrates that our present concept of acute myelomonocytic leukemia should be broadened to include cases in which the leukemic cells possess the morphologic and cytochemical characteristics of both the monocytes and the granulocytes.


Teratogenic action and embryo lethality of AY 9944R. Prevention by a hypercholesterolemia-provoking diet. Embryomortality and teratogenesis provoked by inhibitors of cholesterol synthesis are well demonstrated. Teratogenic action is particularly reflected by holoprosencephalies, but also by uro-genital abnormalities. A hypercholesterolemia-provoking diet has been shown to be completely effective for preventing holoprosencephaly, but only partially so for preventing the uro-genital malformations and fetal mortality. It is thus possible that the two types of abnormalities are governed by different mechanisms. In addition, the diet itself, whose hypercholesterolemic effect is considerable, has certain disadvantages. It seems to have a certain effect on fetal mortality and could be responsible for several uro-genital malformations. This deserves further study.


Interpretation of isolated agenesis of the pituitary. Cholesterol synthesis inhibitors administered to rats caused more or less complete forms of the holoprosencephalic syndrome, consisting of severe abnormalities of the brain, sense organs and pituitary. The absence of the pituitary was also observed in fetuses without externally visible cephalic abnormalities. These observations suggest that the isolated absence of the pituitary is the lesser form of the holoprosencephalic syndrome. This interpretation is also valid for cases of isolated absence of the pituitary observed in humans.


Different teratogenic efficacy to mouse fetal CNS of 5-azacytidine in combination with X-irradiation depends on the sequence of successive application. The single treatment of pregnant mice on day 12 post conception with 5-azacytidine (AzaCr), followed by a single irradiation dose of 200 rad two hours later, is exclusively neurotoxic to the fetus, as shown by a severe hypoplasia of the parieto-occipital regions of the telencephalon. This effect is explicable by the specific function of the mitotic cell population for the integrity of the cortex wall. Combining these two hazards in the reverse manner, i.e., irradiation followed by AzaCr, resulted in no general hypoplastic effect in the forebrain and only caused a depletion of cells in the marginal cortex. This indicates a significantly diminished AzaCr sensitivity of fetal cortical cells subsequent to X-irradiation. In addition, rosette-like cell clustering in the cortex of all X-irradiated animals occurs to a similar degree, irrespective of any additional AzaCr-treatment. The only conformity between these different schedules is that a great portion of the surviving cells is most likely in the DNA synthesizing phase at the time of irradiation. It is therefore concluded that rosette formation starts perferentially from cells injured during the S-phase.


[The inoperable carcinoma of the esophagus: results from radiation therapy and conclusions for irradiation planning (author's transl)]. Conclusions are drawn for irradiation planning from treatment results of 161 patients with inoperable esophageal carcinomas, comparable with reference to the technique applied using moving-field irradiation and delivering a total focal dose of 6000 rd. The palliative effect of radiation therapy is obtained by measurable dilatation of the stenosis in more than 90% of the patients. 5-year survival, however, was found only in cases of an average target volume comprising 500 cm3 within the 80% isodose, largely independent of the stage of the primary tumor. The best correlation between tumor extension and target volume is found in T3 tumors. A target volume larger than 700 cm3 does not seem to be required. The rate and localization of the recurrences are showing that a minimum safety distance beyond the radiologically ascertained tumor expansion is indispensable.


Non-HLA-D determinants detected by the micro-MLC test. A modified, highly sensitive microculture technique was used to demonstrate the presence of ? non-HLA-D-encoded Lad when both stimulator and responder cells were presumably HLA-D identical and when tested in conventional mini-MLC in round bottomed Linbro/Cooke plates failed to demonstrate any significant stimulation.


Specific inhibition of T-cell-mediated cytolosis by alloantiserum against HLA-D-related (DR) antigens. Lymphocytes sensitized against HLA-A and B region-compatible, HLA-D region-incompatible stimulators were cytotoxic for target cells having the correct HLA-D antigens. This form of cytotoxicity was inhibited by platelet-absorbed pregnancy sera containing antibodies to the HLA-DR antigens of the target cells but not by sera with antibodies against other DR antigens. This form of CML was also inhibited by unlabeled monocytes and B cells of the relevant HLA-D specificity but not by T lymphocytes from the same donors.


Role of I-J-subregion gene products in the regulation of macrophage-T-Lymphocyte interaction. Pretreatment of splenic macrophages with anti-Ia serum specific for the various known I subregions and complement deleted the macrophages required for an in vitro antibody response to burro erythrocytes. Antibodies directed at the gene products of the I-J subregion, however, were unique in that they did not require the presence of complement. When macrophages and macrophage depleted cells were obtained from strains B10.A(3R) and B10.A(5R) differing only at the I-J subregion, they could not be mixed in culture for an effective response. Differences at other I subregions using alternative strain combinations resulted in an enhanced response. These results suggest that the I-J subregion has a critical role in macrophage-T cell interaction.


[Dynamic characteristics of the fluorescence of human peripheral blood neutrophilic polymorphonuclear leukocytes and lymphocytes intravitally fluorochrome-stained with acridine orange]. Neutrophil segmentonuclear leukocytes and lymphocytes of the human peripheral blood vitally stained with Acridine Orange (AO) in concentrations of 250 and 330 mcg/ml show different fluorescence dynamics. The number of neutrophil segmentonuclear granulocytes with green fluorescence of nuclei decreases, whereas the number of cells with red fluorescence of nuclei increases. As a criterion of this process, time T1/2 is taken during which the number of green-fluorescent cells decreases twofold. With AO concentrations of 250 and 330 mcg/ml, T1/2 is equal to 40 or 5 minutes, resp. The nuclei of lymphocytes within a 60 minutes observation show green fluorescence. This effect is likely to be due to structural-functional peculiarities of neutrophil segmentonuclear granulocytes and lymphocytes.


[Immunochemical study of the leukocyte myeloperoxidases from various sources]. The antigenic difference between myeloperoxidases of human, rabbit, guinea pig, horse, dog, sheep and mouse leucocytes and horse radish peroxidase was investigated. By counterimmunoelectrophoresis with antiserum specific for human and mouse myeloperoxidase and horse radish peroxidase, the enzyme catalysing peroxidase reaction in leucocytes from the above sources was shown to possess species specificity and different antigenic composition.


Evaluation of the chloride and bromide salts of cetylpyridium for the transportation of sputum in tuberculosis bacteriology. The usefulness of cetylpyridium chloride and cetylpyridium bromide, at a final concentration of 1% in 1% solution of sodium chloride, for the transportation of sputum in tuberculosis bacteriology, was evaluated. It was found that these compounds did not alter the staining properties nor the viability of the tubercle bacilli for at least 14 days.


[Cystectomy as "ultima ratio" after failure of radiotherapy in bladder cancer (author's transl)]. 20 patients were cystectomized an average of 2 years after unsuccessful curative radiotherapy. This technically difficult procedure has a high mortality rate of about 10--20%. None of the patients survived more than 5 years, but improved quality of life was the main subjective impression. A tumor-free specimen -- the effect of radio-induced downstaging -- is no guarantee of long-term survival. 4 of 6 patients without histological tumor cells in the cystectomy or lymph node specimens died of metastic disease. The indications for "salvage" cystectomy were: untreatable bladder hemorrhage, in some instances as emergency; frequently recurrent tumors; and subjectively unbearable state of disease.


[Comparative statistic of complications in openprostatectomy and transurethral resection (author's transl)]. On the basis of the case material of 1049 patients, the advantages and disadvantages of enucleation and the transurethral resection of the prostatic adenoma are compared. There are no significant differences in the lethality of the operation. Although at present preference is generally given to the transurethral resection, this procedure appears less favorable to us with regard to the intraoperative complications as well as late complications of urethral strictures, during long-term follow-up.


Beta-2-microglobulin as renal diagnostic agent. Our group has undertaken the determination of serum levels of beta-2-microglobulin in 250 patients. We have compared this diagnostic entity to radioisotopic glomerular filtration rate, serum creatinine, and blood urea nitrogen. We have also analyzed the cost effectiveness of each of these diagnostic entities and combinations thereof. We conclude that beta-2-microglobulin is a simple test to perform; it is specific, sensitive, and accurate. Its cost effectiveness is excellent. We advocate its wide clinical usage.


Benign prostatic hyperplasia in an XX man. A sixty-nine-year-old white phenotypic male who was being investigated for a myeloproliferative disorder, was found to have an XX karyotype in all cells examined in bone marrow, lymphocytes, and skin fibroblast cultures. Despite essentially no testosterone in the plasma, he also suffered from severe prostatic hyperplasia, a finding not reported previously in patients with this genotype. While endocrine studies showed normal follicle-stimulating hormone, the luteinizing hormone level was twice the upper limit of normal and estrogens were in the normal female range. Except for complete absence of Leydig cells, testicular histology resembled that usually found in Klinefelter syndrome. The patient died of the combined effects of the myeloproliferative disease and urinary tract obstruction. The mechanism of occurrence of the sex chromosome anomaly as well as the cause and implication of the unusual finding of prostatic hyperplasia are discussed.


[Clinico-morphological characteristics of latent form of prostatic cancer]. 36 cases of cancer of the prostate, diagnosed at the histological study of the tissue removed during adenomectomy, have been studied. The most common from is a highly differentiated adenocarcinoma with a well developed parenchyma. Elastic consistency and smooth surface of the tumors make difficult their clinical, diagnosis. As compared to other forms of prostatic cancer, these tumors are characterized by a more favourable clinical course and prognosis.


Antigenic determinants of influenza virus hemagglutinin. II. Antigenic reactivity of the isolated N-terminal cyanogen bromide peptide of A/Memphis/72 hemagglutinin heavy chain. Gel filtration of a cyanogen bromide digest of pure intact hemagglutinin from A/Memphis/102/72 influenza virus allowed the isolation of a variety of fragments. One of these fragments consists of three cyanogen bromide peptides (CNl and CN3 from HA1, and CNl from HA2) which remain linked together by disulphide bonds. This fragment was found to be antigenically active, as it was able to form antigen-antibody complexes (detected by affinity chromatography of radioiodinated peptide-IgG mixtures on protein A-Sepharose) with IgG directed against the protein moiety of viral hemagglutinin. The three cyanogen bromide peptides present in this disulphide-linked fragment were separated by gel filtration, carried out under reducing conditions, and tested for antigenic activity after controlled reoxidation of the individual peptides. Only one cyanogen bromide peptide, CNl from HA1, showed significant binding to antibody. The results indicate that antigenic activity of A/Mem/1O2/72 hemagglutinin resides within the N-terminal 170 amino acid residues of the hemagglutinin heavy chain.


[The diagnostic value and risks of small bowel biopsy: an assessment in 80 patients (author's transl)]. Small bowel biopsy is a routine procedure used mainly in the evaluation of malabsorption. The Crosby-Kugler capsule, which permits the performance of just one biopsy per session and hydraulic biopsy tubes, permitting multiple biopsies from any level of the small bowel, have been in use now with several modifications for about 20 years. The diagnostic range and risks of these tools are described briefly in this paper and discussed in the light of our own experience in 80 patients. In accordance with the experience of others, we found that small bowel biopsy was accompanied by only a few minor complications and we consider it a valuable procedure in clinical diagnosis and experimental research.


[New data on glycosaminoglycans and proteoglycans]. The most important investigations on glycose aminoglycans and proteoglycans, carried out within the recent 5-7 years, are reviewed. Data on primary structures repeating in glycose aminoglycans and on relationship between these biopolymers and protein components in proteoglycans are considered. Newer information on conformational characteristics of glycose aminoglycans, various complexes aggregation of proteoglycans and distribution of the biopolymers in animal tissues and cells is discussed. Properties of proteoglycans in various solutions are briefly considered.


[Therapy of postoperative arrhythmias (author's transl)]. Severe supraventricular and ventricular tachyarrhythmias occurred after surgery in 9 patients. Oxprenolol or pindolol were used as primary drugs in 8 cases. The effect of therapy was documented quantitatively by an arrhythmia computer with trendscription and alarm recording. Beta-receptor blockade was successful in 6 cases. On supraventricular tachycardia with Wolff-Parkinson-White syndrome was abolished by quinidine. Severe ventricular instability with fibrillation following myocardial infarction was temporarily suppressed by lidocaine. One fatal outcome was caused by severe illness. Although all patients were suffering from coronary artery and/or hypertensive heart disease with diminished cardiac reserve, no serious side effects were directly related to beta-receptor blockade. Therefore beta-receptor blocking drugs are recommended as drugs of first choice in the treatment of postoperative arrhythmias induced by sympathetic overstimulation. Dysrhythmias based on cellular pathology should be treated with conventional antiarrhythmic drugs alone or combined with beta-blockers.


[Mechanisms of cardiac rhythm disorder in hypothalamic lesions in basal meningiomas before and after surgery]. Seventeen of 27 patients studied before and after surgical operations for basal meningioma frequently showed forms of cardial sinusal arythmia, sinusal tachycardia and ventricular extrasystolia. These pathological changes in cardiac rhythmics appeared and were aggravated during the postoperational period. Their possible pathophysiological mechanisms, due to the effect of the tumor and operative injury to the hypothalamo-diencephalic apparatus are discussed.


Non-histone chromatin proteins of mouse spleen cells during the primary immune response to sheep red blood cells and aggregated human gamma-globulin. 1. The protein/DNA ratio in chromatin of spleen cells increased during immunization; the ratio was the highest at the time of the maximum antibody synthesis, then decreased to the control values. 2. In the spleen cells of mice immunized with sheep red blood cells or aggregated human gamma-globulin, several characteristic fractions of non-histone chromatin proteins were preferentially synthesized: two antigen-specific fractions were observed in the phase of IgM and IgG synthesis, respectively, and the third, unspecific fraction was detected when the antibody synthesis ceased.


[Receptors for complement on rat mast cells and release of histamine]. Using rat complement-treated zymosan particles a rosetting of purified rat peritoneal mast cells could be demonstrated. The question was investigated whether the binding of activated complement could be a trigger of histamine release. Varying the degree of complement label on the zymosan particles, the time and temperature of incubation and the dependence on Ca2+ ions, we could not induce a release of histamine in any case. The addition of labeled zymosan increased slightly the mediator release induced by ATP. The immunologic significance of the complement receptors on mast cells is still unclear.


[Allergy and lectins: action between IgE-mediated histamine release and glycoproteins from Viscum album L. (mistletoe)]. Low-molecular constituents from Viscum album (toxic) release, in irritatively toxic manner, histamine from human leukocytes without destroying the cells (cytotox test). This histamine release is prevented by preincubation with viscum lectins. The viscum lectin blocks the Fc-portion of allergen-specific IgE against anti-IgE [125I]. Lectin also inhibits the allergen-specific histamine release from human leukocytes occurring through IgE; this can be demonstrated on leukocytes from patients with atopic extrinsic asthma. The inhibitory effect of lectin could also be due to a primary membrane effect, in addition to blockage of IgE.


Macrophage-lymphocyte interaction in Graves' disease and Hashimoto's thyroiditis. The formation of macrophage-lymphocyte rosettes was studied in lymphocyte cultures from patients with Graves' disease; Hashimoto's thyroiditis, other thyroid diseases and control subjects; the cultures were incubated with normal human thyroid and other non-specific antigens. At the end of incubation, the cell pellets were smeared on slides, stained with Wright's stain and the number of rosettes determined under the microscope. The membrane immunofluorescence technique was employed to identify whether the surrounding lymphocytes were T- or B-lymphocytes. In Graves' disease and Hashimoto's thyroiditis, the mean percentages of rosette formation with crude thyroid antigen were 0.98 +/- 0.22% (mean +/- SEM), and 1.15 +/- 0.25%, respectively. These values were significantly higher than those of control lymphocytes (0.03 +/- 0.02%). Lymphocytes from other thyroid diseases also gave higher values than controls. Kidney antigen, used as a control antigen, gave negative results in Graves' disease and other thyroid diseases, but in Hashimoto's thyroiditis, the mean percentage was of borderline significance. In the direct immunofluorescent staining study using fluorescein-conjugated goat anti-human Ig determinants, including the Fab fraction of anti-human IgG, it appeared that both B- and T-lymphocytes were involved in the rosettes, although B-lymphocytes were more numerous. These results indicate that in patients with Graves' disease and Hashimoto's thyroiditis, a probable immune reaction with thyroid antigen can be demonstrated by macrophage-lymphocyte rosette formation.


[Endogenous opiates (endorphins) and pain]. The current knowledge of the physiological role of endogenous opioid peptides and their receptors, is presented. The possible role for pain perception, acupuncture analgesia, stress analgesia and opiate addiction is discussed.


Orthochromatic leukodystrophy with pigmented glial cells. An adult case with clinical-anatomical study. The case history is reported of a woman who died at the age of 36, at the conclusion of 11 years progressive neurological and psychiatric symptomatology. The anatomical and histological examination demonstrated an orthochromatic leukodystrophy with pigmented glial cells. Attention is drawn to the difficulty of finding these cells, which serve to differentiate between the unusual and the "simple" form of the disease. In the reported patient the pigmented cells were found around the vessels and only in specific cerebral locations. It is emphasized that the form is extremely rare (this is the tenth case so far reported). The significance of whether the iron content should be considered as an incidental or necessary finding is discussed. Systematic research for pigmented casts of this kind is taken to be important for all brains presenting a diffuse sclerosis after a protracted clinical course, mainly in adult patients.


A silver impregnation method for reactive microglia in 1 mum epoxy sections. A simple rapid ammoniacal silver nitrate impregnation method for reactive microglia in 1 mum epoxy-embedded tissue is presented. This has been applied to demonstrate argyrophilic microglia in stab-wounded rat brain and in the human cerebellar cortex.


Human corneal endothelium in organ culture. The influence of temperature and medium of incubation. Forty-four human corneas from patients between 21 and 86 years were incubated in Eagle's minimum essential medium with Earle's salts 10--46 h post mortem. The influence of incubation temperature and composition of the medium on endothelial survival was evaluated. Whole corneas were stained by alizarine red. Recent cell loss was indicated by morphological alterations in the endothelial pattern. After 20--28 h of incubation minimum cell loss was found at 31 degrees C when 8% Dextrane-250 and 20% serum or 8% Dextrane-500 and 10% serum was added to the medium.


Postmortal "vital" staining of the external eye. Two hundred and two eyes from 112 individuals were vital-stained from 2 to 46 h after death. Intensity and extension of staining were studied in ten regions. The staining gradually progressed after death, also within non-exposed areas. It most often started anteriorly on the tarsus and finanally included the fornix. Microscopy revealed diffuse cell staining by rose bengal or trypan blue. The cell nucleus was most often more intensely stained than the cytoplasm. Up to five h after death, neutral red had only stained vacuoles in the cytoplasm. Later diffuse staining of cells occurred. Tetrazolium differed from the above dyes in that the pronounced staining seen immediately after death gradually decreased in the course of time. Microscopy disclosed stained inclusion bodies in the cytoplasm up to 18 h after death. Then cell staining was only seen as a rare exception in relation to extracellular dye granules. However, the mucous thread in the inferior fornix showed gradually increasing postmortal tetrazolium staining. The amount of mucus was found to be the same in dead persons as in the living. The characteristic appearance of the dead eye is due, among other things, to ruptures of the corneal epithelium (fluorescein-stained) and cell death, and not to drying up or coating by mucus.


Bleomycin as adjuvant in radiation therapy of advanced squamous cell carcinoma in head and neck. Since 1969, Bleomycin (BLM) has been used in three different ways at the Radium Centre in Copenhagen. First BLM given as the sole treatment led to complete regressions in 12% of 138 patients (12). Secondly BLM was used as simultaneous adjuvant in radiation therapy for 86 previously untreated patients, but 66% developed mucositis which disrupted the treatment. In a third period BLM was therefore combined sequentially with radiation, administered for 2 weeks prior to radiation therapy to 142 patients. The tumour shrinkage achieved with preirradiation BLM was very pronounced in 38% of cases. 104 patients with T3 tumours have been observed for a minimum of 3 years. The medium survival time, 47 months for those sequentially treated patients responding pronouncedly to BLM, was greater than for the remainder of the sequentially treated patients, 16 months (2 p = 0.010) and for the patients treated simultaneously with BLM, 15 months (2 p = 0.013). The prognostic value of the degree of shrinkage achieved with preirradiation BLM treatment is discussed.


Cytology and cytochemistry of acinic cell carcinoma. In a primary material of 314 epithelial parotid tumours treated at the University Hospital in Umeå since 1958, fiften (11 female and 4 male patients) were classified as acinic cell carcinomas. Four of the female patients were in the age range 16--19 years. At the ultrastructural level, granulated cells were the predominant cell type in four tumours studied. Agranulated cells highly reminiscent of intercalated duct cells were also encountered, however. Various cytochemical techniques were employed to demonstrate periodate reactive carbohydrates and acid phosphatase activity. Tumour specimens were also collected and analysed for their content of amylase and cyclic AMP. The cyto- and biochemical findings are discussed and correlated to those observed in normal salivary gland tissue.


Argyrophil cells in the exocrine pancreas. The silver positive cells of the exocrine pancreas and primary pancreatic cancers were studied with the Grimelius silver stain and the Fontana-Masson technique. In the pancreas, cells containing black granules with the Grimelius method, which at the same time react negative to Fontana-Masson, are considered argyrophil. These cells were present in the basal portion of some of the acinar tissue and in the ductal epithelia, as well as in the A cells of islets. The incidence and distribution of these argyrophil cells were also studied in a variety of ductal lesions. In the so-called ductal proliferation numerous numbers of positive cells were found. Argyrophil cells were frequently situated in the basal portion of ductal squamous cell metaplasia. In goblet cell metaplasia the numbers were few, and less than in normal ducts. We concluded that the distribution and incidence of argyrophil cells in the ductal epithelia is related to chronic pancreatitis, and in particular to regenerative processes. The incidence of argyrophil cells in primary pancreatic cancer, excluding islets cell origin, was 18 per 41 cases (43.4%). We considered them pancreatic cancer with argyrophil cells.


Alpha-fetoprotein producing rhabdomyosarcoma of the adult liver. An autopsy case of a rare primary tumor in a 76-year-old man is described. The tumor is histologically comprised of rhabdomyoblasts massively infiltrating without any epithelial elements referable to be as hepatoma. There was no evidence that regarded the hepatic tumor as metastatic secondary tumor in histological examination of other organs. Furthermore, interesting was the production of alpha-fetoprotein from the tumor cells that was detected by immunofluorescent antibody technique. Review of the literature on primary liver rhabdomyosarcomas or adult hepatoblastomas shows no similar case.


Anaphylatoxin-induced shock and two patterns of anaphylactic shock: hemodynamics and mediators. In the dog, different cardiorespiratory reactions were identified in two types of anaphylactic shock and in C5a-AT (anaphylatoxin)-induced shock. All three types had in common a portal blood pooling with consequent decrease in the venous return, cardiac output, and arterial pressure. In anaphylaxis (a) of the first type, at a low titer of hemagglutinating antibodies, the latent period was 68 s and heart and lung function was unchanged. In the second type, at high titer, the latency was 19 s and pulmonary hypertension and decreased heart contractility occurred. After AT injection pulmonary hypertension appeared with tachypnea and unchanged heart function. Tachyphylaxis, but not cross-over tachyphylaxis against the anaphylactic agent and AT was observed in dogs and isolated guinea pig lungs. AT induced a transient release and a, a prolonged release of histamine, prostaglandins (PGs), and thromboxane A2 and endoperoxides from guinea pig lungs. SRS-A was released only in a. Indomethacin inhibited AT-induced release of PGs in guinea-pig lungs and AT-induced hypotension in the dog though it did not prevent the drop in cardiac output. These model studies suggest that different patterns of clinical a. can occur, depending on the type of antibodies and/or mediators involved.


Vascular anatomy and tissue osmolality in the filiform and fungiform papillae of the cat's tongue. The vascular anatomy of the filiform and fungiform papillae of the feline tongue was studied by i.a. injection of India ink. Vascular loops of various appearances were found in the types of papillae studied, i.e. the large and the small filiform papillae and the fungiform ones. Such hairpin loops may function as countercurrent exchangers and to test this hypothesis tissue osmolality was determined in the papillae, while exposing them to various isotonic electrolyte solutions. The large filiform papillae with a vascular arrangement similar to that of intestinal villi exhibited a marked osmolar gradient from tip to base when exposed to a solution containing both glucose and sodium. If sodium and/or glucose was excluded from the solution, tissue osmolality was significantly decreased. This was also the case when the chloride ions of the solution was substituted with sulphate. The small filiform papillae are only provided with one or a few capillary loops. They exhibited a less marked osmolar gradient than the large ones and one of the different electrolyte solutions decreased the gradient. In the fungiform papillae a tissue hyperosmolality at the tip was also demonstrated. It is proposed that the papillary epithelium is provided with active transport mechanism(s) and that the papillary vessels function as countercurrent multipliers. The functional importance of these mechanisms are tentatively discussed.


A case of merbromin (Mercurochrome) intoxication possibly resulting in aplastic anemia. A patient is described who appeared to be suffering from mercury intoxication caused by local application of merbromin to an operation wound and who developed aplastic anemia, which we ascribed to merbromin.


The erythrocyte sedimentation rate and stress. The effect of a 75-hour vigil on the erythrocyte sedimentation rate (ESR), i.a., was studied in two experiments with 63 healthy male volunteers. The ESR was increased at the end of the vigil compared with pre-exposure values. The increases did not correlate significantly with concomitant changes in serum triglycerides, free fatty acids, cholesterol or gammaglobulins, except for a significant, negative correlation with cholesterol changes in one of the two studies. Although the mechanism for the increases in ESR in response to stressor exposure remains unclear, it is concluded that when using the ESR in clinical practice, allowance should be made for situational factors such as the patient having experienced some stressful days and sleepless nights.


Quantitation of coronary arterial narrowing at necropsy in sudden coronary death: analysis of 31 patients and comparison with 25 control subjects. A quantitative analysis of the degree and extent of coronary arterial narrowing by atherosclerotic plaques in the entire lengths of each of the four major coronary arteries in a group of patients dying suddenly from coronary heart disease ("sudden coronary death") is described at necropsy. Cross sections were examined histologically in a total of 1,564 five mm long segments of the left main, left anterior descending left circumflex and right coronary arteries in 31 patients with sudden coronary death, and the observations were compared with those from examination of 1,100 five mm segments of the major epicardial coronary arteries in 25 control subjects. An average of 25 cm (50 five mm segments) of coronary artery were examined from each patient and an average of 22 cm (44 five mm segments) from each control subject. Of the 1,564 five mm segments examined in the 31 study patients, 557 (36 percent) were 76 to 100 percent narrowed in cross-sectional area by atherosclerotic plaques (control subjects 3 percent), 536 (34 percent) were 51 to 75 percent narrowed (control subjects 22 percent), 360 (23 percent) were 26 to 50 percent narrowed (control subjects 42 percent) and only 111 segments (7 percent) had 25 percent or less narrowing (control subjects 33 percent). The amount of severe (greater than 75 percent) narrowing of the right, left anterior descending and left circumflex coronary arteries was similar. Additionally, the amount of severe (greater than 75 percent) narrowing in the distal one half of the right, left anterior descending and left circumflex coronary arteries was similar to that in the proximal halves of these three arteries. The number of 5 mm coronary arterial segments narrowed 76 to 100 percent in cross-sectional area in the 31 study patients was not affected by the patient's age of death, sex, presence or absence of previous angina pectoris or myocardial infarction or the weight of the heart.


Use of prostaglandin E1 in infants with d-transposition of the great arteries and intact ventricular septum. Prostaglandin E1 was used to treat five infants with d-transposition of the great arteries and intact ventricular septum who had persistent severe hypoxemia after the creation of an interatrial communication. Three infants had a dramatic improvement in systemic arterial oxygen saturation associated with dilation of the ductus arteriosus; in two of the three cases urgent surgery was avoided. Two infants had no clinical evidence of increased ductal shunting and no improvement in oxygen saturation. A trial of prostaglandin E1 is recommended for treatment of severe hypoxemia in infants with d-transposition of the great arteries with intact ventricular septum if the presence of a large atrial septal defect is established.


Prognosis for the newborn with transposition of the great arteries. To determine the prognosis for the newborn with transposition of the great arteries, the clinical course of 112 consecutive neonates with dextrotransposition was reviewed. Patients were managed with balloon atrial septostomy at initial cardiac catheterization, palliative operation if needed in the 1st year of life and Mustard's intraatrial baffle repair. The 1st month of life was the period of greatest risk (8 percent mortality rate). Between balloon septostomy and baffle repair, 14 of 103 patients at risk (14 percent) either died or had a cerebrovascular accident. The mortality rate at baffle repair was 14 percent (10 deaths in 71 patients), and there were 3 late postoperative deaths. Actuarial analysis of the data indicates that with this plan of management, approximately 50 percent of newborns with transposition of the great arteries will survive 5 years with excellent function and an additional 15 to 20 percent will survive with one or more medical handicaps.


The utilization of radioimmunoassay antibodies for the immunohistologic staining of polypeptide hormones on paraffin-embedded tissue. A method is described whereby commercially available radioimmunoassay-grade antibodies specific for the polypeptide hormones calcitonin, gastrin, glucagon, and somotastatin are used to detect these antigens on paraffin sections of routinely fixed tissue. The hormone antibodies are applied to deparaffinized tissue sections as the primary specific immune sera using the standard peroxidase technic. The use of these hormone antibodies to detect their respective antigens has proved valuable in demonstrating polypeptide forming tumor cells in pathologic specimens.


Cytochemical diagnosis of acute myelomonocytic leukemia. Blood and bone-marrow smears from adult patients with acute leukemias were stained for esterase reaction, consecutively with naphthol AS D-chloracetate (chloracetate esterase) followed by alpha naphthyl butyrate (nonspecific esterase). The two substrates were, respectively, granulocyte- and monocyte-specific. By this method three subgroups of acute nonlymphocytic leukemias could be distinguished. Leukemic cells may be positive for either chloracetate esterase or nonspecific esterases, and the authors believe these two subgroups represent "true" granulocytic and "pure" monocytic leukemias. In a third group, leukemic cells contained both esterases in the same cell, and it is believed this group may represent "true" myelomonocytic leukemias. In the majority of patients in this group, leukemia evolved from a preleukemic phase. When only Romanowsky-stained smears are used, the monocytoid feature and absolute elevated monocyte counts in acute granulocytic leukemia may lead to an erroneous diagnosis of the leukemia  as acute myelomonocytic leukemia. This happened in five of the 13 cases in the study. The presence of granulocyte- and monocyte-specific esterases in a single cell supports the concept of a common origin of granulocytes and monocytes. The authors conclude that the combined esterase reaction can distinguish among acute granulocytic leukemia, acute monocytic leukemia, and acute myelomonocytic leukemia.


Comparison of stainable iron in aspirated and needle-biopsy specimens of bone marrow. A comparison of stainable iron in simultaneously obtained aspirated smears and needle-biopsy specimens from 1,000 patients was undertaken. Significant differences occurred when iron was assessed as absent in the aspirated smear. In only 35% of the corresponding needle-biopsy specimens was iron absent. When only the aspirated smear was used, there was a significant overdiagnosis of iron deficiency. In general, iron tended to be less in the aspirated smear; however, correlations were better when iron stores were assessed as being present or increased in the aspirated smears, for stainable iron in the needle-biopsy specimen was always present in equal or greater amounts. Hemosiderotic smears (increased stainable iron) and needle-biopsy specimens (3+ -4+) correlated well. The aspirated smear and needle-biopsy are complementary procedures, and each has advantages. In the authors' experience, the needle-biopsy was preferable to the aspirated smear for evaluation of iron stores, particularly when iron stores were low or absent.


Pneumocystis carinii pneumonia. An approach to rapid laboratory diagnosis. Laboratory procedures used to establish the diagnosis of Pneumocystis carinii pneumonia were evaluated using an experimental murine model. Touch preparations and suspension smears were prepared from lung tissue know to contain Pneumocystis cysts. These preparations were stained by a variety of methods known to demonstrate either cyst forms or sporozoites and trophozoites. Suspension smears proved to be superior to touch preparations in terms of cyst content and homogeneity of staining. Also, methods that stain cyst forms were superior to those that stain sporozoites and trophozoites for location and identification of organisms. The authors believe that suspension smears prepared from lung tissue and stained with toluidine blue O should be examined initially as a rapid screening method for Pneumocystis cysts. When the results of this initial screen are negative or inconclusive, additional suspension smears stainded by the modified Gomori methenamine silver nitrate method should be examined, pending availability of histologic sections.


Genetic polymorphism of the A subunit of human coagulation factor XIII. Utilizing a fluorescent technique for the localization of transglutaminase activity after electrophoresis on thin layer agarose gels, we observed a new polymorphism of coagulation factor XIII in both platelets and plasma. The electrophoretic pattern was that of a dimeric protein. Homozygotes gave a single band, while heterozygotes presented a three banded pattern. The polymorphism was found to be due to variation of the A subunit. Data from Australian blood donors indicate that the A subunit of factor XIII has an autosomal locus.


Research needs in low vision.

Research needs in low vision. Research needs are presented for several areas of low vision: (1) reading and learning, (2) new low-vision aids, (3) modifying the environment, (4) utilization of disciplines, (5) new technologies, and (6) integrating low-vision services.


Peroperative fat staining of frozen sections in primary hyperparathyroidism. Roth and Gallaher recently described a fat staining method for rapid peroperative differentiation between parathyroid adenoma and chief cell hyperplasia. They used Sudan IV in a solution of ethanol and acetone. This solution, however, was found to cause a considerable dissolution of small lipid droplets from the tissue; in our hands sections stained with this technique were diffcult to interpret. To diminish the loss of fat from the tissue, we have used a modification of Lillie's supersaturated ispropanol method with oil red O. This method gave a deeper staining and increased the difference between hyperfunctioning and unnivolved parathyroid tissue with respect to the amount of stainable lipid in the chief cells. It was found to be a valuable supplement, adding a functional dimension to the structural interpretation of the tissue, and it facilitated the peroperative distinction between ademona and hyperplasia. The pattern of lipid distribution within the glands from patients with nodular hyperplasia suggests that the compact nodules of such glands are autonomously hyperfunctioning, whereas the intervening parts of the parenchyma are more or less responsive to the serum calcium level.


Reconstruction after total gastrectomy: construction of a Hunt-Lawrence pouch using Auto Suture staples. A simple gastric reservoir using autosuture staples can be constructed rapidly after total gastrectomy. The importance of both a reservoir capacity and delayed evacuation was supported by the clinical course of most of the patients. All 18 patients had improved nutrition. Patients resected for cure tended to have greater weight gain and less abdominal discomfort. Occasionally patients who underwent only palliative resection were long-term survivors with a clinical course similar to that of patients resected for cure. On the basis of these results, it is unfair not to consider this type of reconstruction in patients who have undergone a total gastrectomy because it adds little to the operating time, yet it provides better palliation in terms of nutrition and abdominal comfort on both a short- and long-term basis.


Binding of antigen to surface Ig during in vitro hyposensitization of isolated rat mast cells. The mechanism of in vitro hyposensitization was examined in isolated rat mast cells. Surface distribution of immunoglobulin was examined by immunofluorescence technique. Hyposensitization could be ascribed neither to changes in the distribution of surface immunoglobulins nor to capping or shedding phenomena. The distribution of cell-bound antigen and the number of antigen-positive cells (patch-bearing cells) was not changed by hyposensitization. The binding of antigen to surface Ig was quantitated by I125-labelled antigen. In rats sensitized to two different antigens, hyposensitization of the mast cells with one of these inhibited the subsequent histamine release induced by either of the two antigens, whereas no changes were observed in the antigen binding capacity. The present investigation indicates that the mechanism of in vitro hyposensitization is the result of events secondary to the initial binding of antigen to the mast cell surface immunoglobulin.


Audiotape slide teaching: use with automated and simple apparatus. Using a cross-over design trial and a series of 20 audiotape slide programmes on clinical physics and measurement, no difference in teaching effectiveness was found between their use with synchronised automated apparatus and simple manual equipment. In both cases 91% of the main lecture points were immediately recalled and there was a significant gain in knowledge from an average pre-test MCQ score of 43%--76% post-test score. The advantages and disadvantages of automated and simple equipment were discussed. The anaesthetists found the teaching by the tapeslide technique very acceptable, helpful and useful.


The cerebellar projection from the lateral reticular nucleus as studied with retrograde transport of horseradish peroxidase. The cerebellar projection from the lateral reticular nucleus (NRL) was studied in cats by means of retrograde axonal transport of horseradish peroxidase (the projection to the paramedian lobule was not included, see Brodal, 1975, for afferents to this cortical region). The entire cerebellar cortex and all cerebellar nuclei receive fibres from the NRL. The strongest connection is with the anterior lobe and lobulus VIIIB of the posterior lobe vermis. As concerns the anterior lobe the observations confirm the previous finding by Brodal (1975) that there is a clearcut topical pattern in the nuclear projection to this part of the cerebellum. The observations furthermore show that crus II is the only cerebellar region devoid of fibres from the subtrigeminal part of the NRL. The cerebellar projection from the NRL is bilateral with a heavy ipsilateral preponderance. The large majority of the labeled cells within the NRL are of the small category (less than 25 micrometer in size). This and the other findings are discussed in relation to previous studies on the efferent and afferent connections of the nucleus.


Spinal ganglion cells innervating the stomach of the rat as demonstrated by somatopetal transport of horseradish peroxidase (HRP). Spinal ganglion cells innervating the stomach of the rat were demonstrated using the somatopetal horseradish peroxidase (HRP) transport technique. After injection of the tracer into the anterior wall of the stomach, labelled neurons were observed bilaterally within spinal ganglia T4--L1. They were most numerous in ganglia T8--T10. No clear evidence of somatotopic organization of the labelled cells was found. Most of the cells measured 35--40 micrometer in diameter. There was a striking paucity of cells smaller than 30 micrometer. The predominance of medium-sized spinal ganglion cells may indicate that the stomach is innervated mainly by small myelinated rather than unmyelinated splanchnic afferent fibers.


Lidocaine (without epinephrine) does not affect the fine structure or microtubules of the trigeminal nerve in vivo. The authors examined the fine structure and microtubules of unmyelinated axons and Schwann cells in the infraorbital branch of rat trigeminal nerve 1, 3.5 and 24 hours after intraneural injection of lidocaine HC1, 1--4 per cent, 0.2 ml, or saline solution, 0.2 ml; untreated control nerves were also examined. These concentrations of lidocaine are more than sufficient to block impulse conduction and rapid axonal transport in rat infraorbital nerve, but contrary to previous reports, significant structural change was not found as compared with control nerves.


A novel method to assess antihistamine and decongestant efficacy. A method has been developed to equalize as many of the variables as possible in a clinical trial to assess the response to medication in allergic rhinitis. This method was used to study the effectiveness of azatadine maleate and pseudoephedrine sulfate alone and in combination. Azatadine effectively relieved symptoms of hay fever but did not reduce nasal congestion to a significant degree. Pseudoephedrine relieved nasal congestion but did not reduce symptoms. The combination of azatadine and pseudoephedrine relieved both symptoms and congestion.


Denaturization of allergen P: effect on allergenicity, antigenicity and immunogenicity. When allergen P was denatured by 8M urea, the modified molecule still reacted with IgE specific for the native allergen but not with hemagglutinating antibodies. Heating at 100 degrees C abolished the reaction in both cases. The results suggested differences between allergenic and antigenic capacities which may be based on structural differences of the antigenic determinants.


Immunologic diagnosis of cancer.

Immunologic diagnosis of cancer. Immunologic tests for the specific detection of tumor specific antigens would provide a method for the early detection of malignancy. Using immunodiffusion and immunofluorescent techniques it is possible to measure antigenic markers. Carcinoembryonic antigen and alpha1 fetoprotein are helpful in following treatment and prognosis of a patient. Skin tests and tests for cellular immunity help to measure the immune competence of the cancer patient.


[Use in histological technic of naphtol AS-D chloroacetate for demonstration of granulocyte series elements]. The authors report their experience of a histo-enzymatic method, using Naphtol AS-D Chloroacetate as a substrate, for the specific demonstration of elements of the granulocyte series, in histological sections, after formol fixation and mounting in paraffin wax. This simple and reliable staining technique offers the possibility of very rapid diagnostic orientation in haematological conditions.


A study of HLA-A, B, C, and DR specificities in pigeon breeder's disease. The frequencies of HLA-A, -B, and -C antigens were determined among 51 symptomatic pigeon breeders, 102 asymptomatic pigeon breeders, and 100 normal blood donors. The HLA-DR specificities were also studied in 32 symptomatic and 29 asymptomatic pigeon breeders. All subjects were white. Symptomatic subjects were defined by the development of respiratory symptoms or decreased pulmonary function after aerosol challenge with pigeon serum. Asymptomatic subjects were comparably exposed to pigeons but had no pulmonary signs or symptoms after aerosol challenge. No significant association was found between any of the tested HLA specificities and pigeon breeders. This lack of association and the observed paucity of multiplex families indicate that HLA complex genetic factors tested in this study do not favor the development of pigeon breeder's disease in exposed persons.


An animal model of hypersensitivity pneumonitis in rabbits. Development of chronic pulmonary inflammation and cell-mediated hypersensitivity after repeated aerosol challenge. Chronic pulmonary inflammation was produced in immunized rabbits by repeated aerosol exposure to soluble antigen. The pulmonary inflammatory response was correlated with the development of cell-mediated hypersensitivity in the lung as evaluated by migration-inhibition studies using bronchoalveolar cells. Such inflammation could be produced with either pigeon dropping extract, an etiologic agent of hypersensitivity pneumonitis, or with human gamma globulin. Development of the inflammatory response was immunospecific and could not be transferred to normal recipients with large quantities of immune serum. Collectively, these data suggest that the development of pulmonary inflammation was due to a cell-mediated immunologic reaction in the lung.


Pharmacokinetics of halogenated hydrocarbons. Pharmacokinetic modeling is being applied to chemicals posing health hazards because of their presence as contaminants in our environment. This paper presents a rationale for constructing mathematical models based on anatomic and physiological principles. Pharmacokinetic investigations incorporating such models are illustrated for two halogenated hydrocarbons that exhibit different patterns of disposition in the rat: 2,2',4,4',5,5'-hexachlorobiphenyl and Kepone. The models permit interspecies comparison and prediction of normal pharmacokinetic behavior and alterations induced by intervention measures. As an example a theoretical limit is predicted in the case of a treatment for Kepone poisoning involving oral administration of adsorbents.


Ethical considerations in cancer chemotherapy. Cancer treatment has improved dramatically in the recent past. As a consequence, the range of choices has been greatly broadened. With a much larger range of choices and a larger range of treatment disciplines available, decisions regarding treatment create major ethical burdens for the physician and for the patient. The participation of the patient in such choices improves the ethical posture of the therapist, whenever that is possible, and also improves the probability that the best treatment will be administered. Clinical research plays an important role in cancer treatment, not only in the the discovery of new treatment, but in the provision of the best possible care for patients participating in such research. At all levels of clinical research, such treatment has the potential for being the best available treatment when conducted in the most professional manner. While poor clinical research, like poor conventional treatment, certainly exists, it is nonetheless true that clinical research has a permanent place in cancer treatment and provides an important alternate to cancer quackery. Virtually every patient with a malignant disease has some effective treatment available to him, even in the most hopeless circumstance, where the Phase I type of clinical investigation still offers significant opportunity and hope.


Experience with a hospice-care program for the terminally ill. A hospice-care program offers an opportunity to provide effective palliative care for patients terminally ill with malignant disease and to develop improved methods for coping with the problems of the dying patient. All patients for whom antitumor therapy does not offer a reasonable possibility of cure are eligible for Church Hospital's multidisciplinary program, the focus of which is on both the patient and his family. Acceptance by medical staff, patients and families has been enthusiastic. Both conventional and unconventional methods can be helpful in making terminally ill patients more comfortable. Much has been learned about the control of pain in such patients. Intestinal obstruction can often be managed non-operatively without the use of nasogastric tube. Other common symptoms such as weakness, anorexia, depression, dyspnea, etc. can be relieved with varying degrees of success. An objective of the program is to allow the patient to be at home for most of his terminal illness and to die there if possible. By utilizing patient and family instruction, visiting nurses and home health aides, approximately two-thirds of the patients in the program at any given time are at home. Basing the program in an acute care hospital has allowed coordination with the curative treatment of malignant disease and effective use of radiation and chemotherapy for palliative purposes. The organizational structure, financing, facilities and clinical experience with 100 consecutive patients of the Church Hospital hospice-care program are described.


[Hormonal desexing of boars with chlormadinone acetate]. Chloromadinone acetate produces a dependable desexualising effect on boar by contant administration in feed rations of 30 mg per die over 70 days. Sexual odour thus can be widely eliminated. Other aspects studied in a group of 107 boars are body weight development, sexual behaviour, slaughter yield, and skin quality.


An investigation of elevated blood lead levels in Detroit children. The findings of this report are based on data from 1309 children living near three heavily traveled streets in Detroit. This study was designed to determine the most probable reasons for elevated blood lead levels in young children. The authors used a linear multivariate regression to evaluate the effects of distance from the highway, age and sex of the child, and housing condition. The authors found that the distance the child lived from the highway did not have a measurable affect on blood lead level. Housing condition played the most important role in determining blood lead level. Poorer housing caused a higher average blood lead level and caused the blood lead distribution to skew, giving a significantly greater number of blood leads over 40 micrograms/100 ml. Age also was important in determining blood lead level. There was a decrease of about 1 micrograms/100 for each increase of one year in the child's age. Sex also appeared to play a small role. In all three areas of the study, blood lead levels were slightly higher in girls than in boys. This study shows, as many have before, that the primary cause of the lead problem with children can be greatly diminished by improved housing.


Acridine orange particles in cultured fibroblasts. A comparative study of macular corneal dystrophy, systemic mucopolysaccharidoses types I-H and II, and normal controls. Vital staining with the fluorescent dye, acridine orange, was evaluated as a means of detecting abnormalities of lysosomes in cultivated fibroblasts of patients with macular corneal dystrophy and mucopolysaccharidoses types I-H (Hurler's syndrome) and type II (Hunter's syndrome). Multiple cultures were compared with normal fibroblasts using a "double-masked" design to exclude observer bias. Cells of patients with the mucopolysaccharidoses were easily and accurately separated from other fibroblasts. Contrary to a recent report, corneal fibroblasts of patients with macular corneal dystrophy were indistinguishable from control cells.


Automotive exhaust and mouse activity: relationships between pollutant concentrations and decreases in wheel running. Groups of male and female mice inhaled either clean air, 100 ppm carbon monoxide, or light-irradiated and nonirradiated automotive exhaust containing nominally 25, 50, 75, or 100 ppm carbon monoxide in three tests with exposure lasting from 4 to 7 days. Exhaust from a factory or lean-tuned engine in the first and third tests reversibly suppressed activity wheel running during exposure in mice of both sexes by as much as 78.3 and 83.1%, respectively. Light-irradiated exhaust suppressed running more than nonirradiated exhaust. For the second test, when the engine was tuned to be low in pollutants other than carbon monoxide, exhaust did not suppress running. Exposure to carbon monoxide alone only slightly decreased running in male mice, but increased running in female mice.


A comparative study of O'nyong nyong virus with Chikungunya virus and plaque variants. Two plaque variants of Chikungunya (CHIK) virus were serologically compared with O'nyong nyong (ONN) virus in order to elucidate the reported one way antigenic relationships between the two viruses. Three different hypotheses are examined and evidence is shown to support one of them. Comparison of some biological properties of the viruses showed ONN to be distinct in some respects. All viruses and variants were found to replicate in Anopheles gambiae cells.


Inosiplex in the treatment of subacute sclerosing panencephalitis. Six patients with subacute sclerosing panencephalitis were treated with the antiviral agent inosiplex. The clinical condition of four children continued to deteriorate in spite of the treatment, the condition of one remained unchanged, and that of another improved minimally. The effect of inosiplex in these patients is not evident since this group demonstrated a pattern of progression similar to that seen in subacute sclerosing panencephalitis without specific treatment.


Differential effect of clofibrate on inflammation-induced alterations in plasma proteins in the rat. Daily intramuscular injections of clofibrate begun 6h before the initiation of inflammation induced by the subcutaneous injection of turpentine exerted a differential, dose-dependent inhibition of the anticipated acute-phase globulin response. Specifically, clofibrate at 140mg/kg muted the increase in alpha(2)-macrofoetoprotein, but did not affect that of seromucoid or haptoglobin and only transiently inhibited the rise in copper and the rebound in transferrin. A higher dose, 280mg/kg, markedly suppressed alpha(2)-macrofoetoprotein appearance and the rebound in transferrin, somewhat inhibited the increase in seromucoid and haptoglobin and only transiently affected the rise in plasma copper; 420mg of clofibrate/kg very nearly abolished the appearance of alpha(2)-macrofoetoprotein, markedly suppressed the transferrin rebound and the increases in seromucoid and haptoglobin and again only transiently affected the increase in copper. Clofibrate did not diminish the localized inflammatory response, did not cause microscopically detectable liver damage and did not prevent the hypozincaemia, hypoalbuminaemia and enhanced amino acid uptake by liver usually associated with inflammation. Thus it is unlikely that clofibrate exerted its dose-dependent selective inhibition by muting the initial stimulus or by impairing hepatic metabolism. This seemingly selective action of clofibrate on plasma-protein alterations during inflammation may provide a means of elucidating the function of individual acute-phase globulin during disease. Clofibrate of itself, apart from inflammation, produced decreases in plasma zinc, copper, transferrin and seromucoid and an increase in hepatic amino acid uptake that were to some extent dependent on the dose of the drug.


Evidence for a role of phosphatidylinositol turnover in stimulus-secretion coupling. Studies with rat peritoneal mast cells. Histamine secretion and phosphatidylinositol turnover were compared in antigen-sensitized rat peritoneal mast cells stimulated with a number of different ligands. A small and variable increase in the incorporation of [(32)P]P(i) and of [(3)H]inositol into phosphatidylinositol was observed when the cells were treated with immunoglobulin E-directed ligands (antigens and concanavalin A), and this was accompanied by a low amount of secretion (<10% of total cell histamine). In the presence of added phosphatidylserine, the addition of immunoglobulin E-directed ligands invariably led to an enhanced rate (approx. 4-fold) of labelling of phosphatidylinositol and, in the presence of Ca(2+), this was accompanied by the secretion of histamine. The labelling of phosphatidylinositol and histamine secretion were also stimulated by chymotrypsin and compound 48/80. Whereas the phosphatidylinositol response did not require the presence of extracellular Ca(2+), the secretion of histamine was either enhanced or dependent on extracellular Ca(2+) (depending on the ligand used). The dependence on ligand concentration for the phosphatidylinositol response and histamine secretion were similar. The increased isotopic incorporation into phosphatidylinositol continued for about 1h although histamine secretion (elicited with concanavalin A) stopped within 2min. These results support the proposition that metabolic events involving phosphatidylinositol play a necessary intermediate role in the regulation of Ca(2+) channels by ligand-activated receptors.


Binding of unmodified low-density lipoproteins to human fibroblasts. An investigation by immunoelectron microscopy. The bindinf of unmodified low density lipoproteins to the plasma membrane of fibroblasts was studied at the ultrastructural level. The bound low density lipoprotein was visualized by an indirect immunoperoxidase technique, with the use of an antiserum against apoprotein B. Immunoreactive regions representing bound apoprotein B were found on the plasma membrane, in indented regions with a diameter of 0.15--0.30 micrometer and a fuzzy coat on the cytoplasmic side. Fibroblasts from a patient homozygous for hyperlipoproteinaemia type IIa showed no immunoreactive material in the indented regions. The specific 125I-labelled low density lipoprotein binding to these homozygous fibroblasts was 7% compared to control fibroblasts.


Association of myelin basic protein with detergent micelles. Equilibrium measurements of the binding of central nervous system myelin basic protein to sodium dodecyl sulphate, sodium deoxycholate and lysophosphatidylcholine have been obtained by gel permeation chromatography and dialysis. This protein associates with large amounts of each of these surfactants: the apparent saturation weight ratios (surfactant/protein) being 3.58 +/- 0.12 and 2.30 +/- 0.15 for dodecyl sulphate at ionic strengths 0.30 and 0.10, respectively 1.34 +/- 0.10 for deoxycholate (at 0.12 ionic strength) and 4.0 +/- 0.5 for lysophosphatidylcholine. Binding to the ionic surfactants increases markedly close to their critical micelle concentrations. Sedimentation analysis shows that at 0.30 ionic strenght in excess dodecyl sulphate the protein is monomeric. It becomes dimeric when the binding ratio falls below 1 at a free detergent concentration of approximately 0.25 mM: below this concentration much of the protein and deterent forms an insoluble complex. The amount of dodecyl sulphate bound at high concentrations and at both above-mentioned ionic strengths corresponds closely to that expected for interaction of a single poly-peptide with two micelles. Variability of deoxycholate micelle size on interaction with other molecules precludes a similar analysis for this surfactant. Association was observed only with single micelles of lysophosphatidylcholine. The results provide strong evidence for dual lipid-binding sites on basic protein and indicate that lipid bilayer cross-linking by this protein may be effected by single molecules.


[Quantitative immunoenzyme technic in the evaluation of specificity of the beta 1-g-globulin test in trophoblastic tumors]. The method of guantitative immunoenzymatic determination of beta 1-G-globulin (TSG) in the blood serum has been developed. The sensitivity of the method is 6 ng/ml TSG. It has been shown that elevated levels (12-100 ng/ml and higher) are usually observed in trophoblastic tumours of the uterus. The TSG ectopic synthesis is found to proceed in some tumours of the gastrointestinal tract and testicular teratoblastomas.


[Simple method of detection of Fc receptor sites on cells]. A bacterial adhesion technique for identification of Fc-receptor bearing cells is based on the ability of Staphylococcus aureus, Cowan 1 strain, to form rosettes with cells previously treated with immune complexes or heat aggregated Ig G. The new technique is distinguished from other Fc-receptor detection methods by its simplicity and ease of detection of both immune complexes and aggregated IgG-binding cells.


Immunologic studies of native and modified human factor VIII/von Willebrand factor. Factor VIII/von Willebrand factor (FVIII/vWF) is a glycoprotein with a molecular weight greater than one-million daltons. Two activities are associated with this large molecule: FVIII procoagulant activity and vWF activity. Incubation of FVIII/vWF with proteolytic enzymes causes rapid inactivation of the FVIII procoagulant activity but has little effect on the vWF activity or antigenicity. In an attempt to gain insight into the structural features required for these two activities, antisera were raised in rabbits to normal, thrombin-inactivated, and plasmin-inactivated FVIII/vWF. All of these proteolytically modified forms of FVIII/vWF cross-reacted with each of the rabbit antisera; each blocked the ability of a human inhibitor to inactivate native active FVIII/vWF. Each of the antisera was a potent inhibitor of vWF activity and inactivated vWF activity at the same titer. The antisera were much less potent inhibitors of FVIII activity than of vWF activity. Antibodies to thrombin-inactivated FVIII/vWF or normal FVIII/vWF had about the same ability to inactivate FVIII procoagulant activity. Surprisingly, those to plasmin-inactivated FVIII/vWF still retained about 50% of this inhibitory capacity. A comparison of the three types of antigens by polyacrylamide gel electrophoresis in sodium dodecyl sulfate-6 M urea demonstrated that the structure of thrombin-inactivated FVIII/vWF was indistinguishable from that of normal FVIII/vWF, while plasmin-inactivated FVII/vWF was completely cleaved to lower molecular weight fragments. Some of the reported variations in the ability of rabbit antibodies to inhibit procoagulant activity may be due to partial degradation of the starting antigen. The retention by FVIII/vWF protein of its immunologic properties even after extensive proteolytic degradation suggests that under nondenaturing conditions, the conformation of the native and degraded molecules are very similar.


[Anti-smoking activities in Switzerland and their evaluation]. Activities against smoking in Switzerland are planned and evaluated by the "Swiss Interagency Council on Smoking and Health". With special regard to the particular local situation it tries to coordinate the programs of its members (private organization as well as state authorities) in accordance with a multi-step plan (information about the effects of smoking, publicity, motivation for change of behavior, support of smoking-withdrawal programs, influencing legislation). Evaluation of anti-smoking programs so far has only been fragmentary, showing some circumstantial evidence for a trend towards non-smoking. It is planned to evaluate the impact of future programs by periodical surveys of representative samples of the adult population, monitoring knowledge of the hazards of smoking, attitudes towards smoking, motivation for change of behavior as well as the actual smoking habits.


The cytological differentiating potential of pineal parenchymal neoplasms (true pinealomas). A clinicopathological study of 28 tumours. A series of 28 pineal parenchymal tumours is described, with special reference to the potential of some of these neoplasms to differentiate along glial or ganglionic lines, or both. The more undifferentiated tumours (pineoblastomas, 11 cases) were the most frequent: they are histologically similar to medulloblastomas. One example showed focal differentiation to retinoblastoma at the primary site. The histological features of pineoblastomas merged with those of pineocytomas (7 cases), in which the lobular architecture is reminiscent of that of the mature pineal gland. In addition, 10 further examples in the group of pineocytomas showed more advanced differentiation as follows: towards astrocytes only (2 cases), towards ganglion cells only (1) case) and towards both astrocytes and ganglion cells (gangliogliomas) (7 cases). Confirmation of the pineal parenchymal nature of these neoplasms and of their differentiating potential was provided by a modification of the Achúcarro-Hortega's silver carbonate impregnation technique for pineal parenchymal cells, by specific silver impregnations for axonal processes, and by an immunoperoxidase stain for glial fibrillary acidic (GFA) protein. Electron microscopy of one new example of pineocytoma with neuronal and astrocytic differentiation demonstrated the presence of numerous microtubules, of clear-centred and dense-core vesicles, and of synaptic complexes. Seven illustrative clinical histories with pathological findings are presented. The identification of special features of cellular differentiation is of importance in evaluating the biological behaviour of these neoplasms since a definite correlation can be established between the patient's age, some of the cytological variants, and the malignant potential of the tumour. Pineoblastomas are highly malignant neoplasms of children and young adults which disseminate widely throughout the cerebrospinal fluid pathways. -ineocytomas without cellular evidence of further differentiation occur at any age and are also clinically malignant, but with a somewhat lesser tendency to metastasize than pineoblastomas. Pineobytomas with astrocytic differentiation occur in adults and may be either slowly growing or malignant. Pineocytomas with neuronal or with neuronal and astrocytic differentiation occur in later life, remain localized, and are relatively benign. Since the latter account for approximately one-third of pineal parenchymal tumours and are likely to be relatively radio-resistant, tissue diagnosis is imperative for a determination of the therapeutic approach. Radiation to the entire neuraxis should be administered to patients with pineoblastomas and malignant pineocytomas in view of their high frequency of cerebrospinal metastasis. An accurate histological classification of these tumours therefore carries important clinical and therapeutic implications...


A comparison of the organization of the projections of the dorsal lateral geniculate nucleus, the inferior pulvinar and adjacent lateral pulvinar to primary visual cortex (area 17) in the macaque monkey. Both anterograde and retrograde transport tracing methods were used to study the organization of the projections of the dorsal lateral geniculate (DLG), the inferior pulvinar and subdivisions of the lateral pulvinar to primary visual cortex (striate cortex or area 17). The DLG projects only to striate cortex. These projections are retinotopically organized, and do not extend to any cortical layers above layer IVA. In contrast the inferior pulvinar (PI) and the immediately adjacent portion of the lateral pulvinar (PL alpha 48) project to both striate and prestriate cortex. The projections from these two thalamic areas to the striate cortex are also retinotopically organized and exist in parallel with those from the DLG. In contrast to the DLG, the projections from PI and PL alpha terminate above layer IVA in striate cortex, i.e. layers I, II and III. In prestriate cortex the layers of termination include layers IV, III and I. The pulvinar terminations in layers II and III of area 17 occur in segregated patches as do the geniculate terminations in layers IVC and IVA. On the other hand the pulvinar terminations in layer I which overlie those in layers II and III of area 17 appeared to be continuous. Control studies show that the remainder of the lateral pulvinar overlying PL alpha does not project to striate cortex. It is concluded that there are 3 visuotopically organized inputs from the lateral thalamus to primary visual cortex and that each of these inputs have different layers of termination. The inputs from PI and DLG can convey direct retinal inputs while those from PI and PL alpha can also be involved in intrinsic cortico-thalamocortical connection with prestriate cortex. It remains, then that it cannot be tacitly assumed that the ascending inputs which influence the response properties of the primary cortical neurons arise solely from the dorsal lateral geniculate nucleus. It is also argued that these inputs to the supragranular layers may be excitatory as those from the DLG to the IVth layer.


Specific uptake and retrograde flow of antibody to dopamine-beta-hydroxylase by central nervous system noradrenergic neurons in vivo. This study sought to determine whether the administration in vivo of antibody to dopamine-beta-hydroxylase (AD beta H) is taken up by central noradrenergic neurons and transported by retrograde flow to the cell bodies of origin. AD beta H serum or preimmune serum (control) in volumes of 1--20 microliter were stereotaxically injected into the lateral ventricle. Rats were sacrificed at times ranging from 1 h to 8 days. Cryostat sections were stained with fluorescein conjugated IgG. After 24 h, a bilateral granular fluorescence was seen only in neuronal cell bodies corresponding to noradrenergic cell groups A1--A7 with the most intense fluorescence localized within perikarya and processes of the locus coeruleus (A6) and subcoeruleus. This technique also permitted the visualization of the ascending dorsal and ventral noradrenergic bundles as well as varicose fibers and terminals in a pattern identical to that reported with histofluorescence, autoradiographic, biochemical and classical immunofluorescence techniques for the identification of noradrenergic fiber distributions. At 3 and 6 h, the first detectable fluorescence was observed in forebrain noradrenergic terminals and in fibers of the dorsal and ventral noradrenergic bundles. At 10 h fluorescent varicosities were first visualized within the caudal dorsal bundle and some cytoplasmic fluorescent particles were seen within locus coeruleus cell bodies. After 18 h locus coeruleus and subcoeruleus cell bodies were heavily stained, whereas medullary noradrenergic cell groups and nerve fibers were not labeled until after 24 h. An intense locus coeruleus fluorescence remained for 3 days and was completely absent after 6 days. Bilateral transection of the dorsal noradrenergic bundle in the rostral mesencephalon, at the time of injection, effectively blocked the retrograde transport of fluorescing material to the locus coeruleus. The overall staining pattern suggests that, in vivo, central noradrenergic fibers are capable of taking up antibody to dopamine-beta-hydroxylase. The ability of a dorsal bundle transection to abolish locus coeruleus staining, as well as the time course of AD beta H staining in noradrenergic neurons, suggests that AD beta H is transported via a rapid retrograde flow process. This technique combines retrograde transport of a marker protein with the sensitivity and specificity of immunocytochemical procedures to provide a new tool for the neuroanatomical study of neurotransmitter systems.


Nerve terminal proteins of the rabbit visual relay nuclei identified by axonal transport and two-dimensional gel electrophoresis. The proteins in nerve terminals can be uniquely identified by two-dimensional gel electrophoresis of proteins labeled during synthesis in the cell body and then transported intra-axonally to the terminals. We have explored the potential of the identification procedure by comparing the proteins which are transported from the retina to the lateral geniculate nucleus (LGN) and the superior colliculus (SC) of the rabbit. We have been able to identify between 150 and 200 proteins which ate common to both LGN and SC nerve terminals, very few of which are present at significantly different concentrations in one nucleus relative to the other. The similarity between proteins sent from the retina along two neural pathways subserving different functions illustrates the subtlety of biochemical changes that must underlie physiological differences. Only a small fraction of the labeled proteins are major proteins of the relay nuclei as judged by Coomassie-staining, and some of these arise from in situ nonspecific labeling with blood-borne radioactivity, rather than by transport to the terminals. We have shown that about 5 times more proteins are transported at fast than at intermediate transport rates. More than 50% of the fast proteins turn over rapidly and are gone in 24 h. Few intermediate proteins turn over rapidly. Since only 6% of the proteins in the relay nuclei (at 36 h) could not be detected in the optic tract at that time, transsynaptic labeling by breakdown and resynthesis must be small, if it occurs at all.


Primary osteogenic sarcoma: the rationale for preoperative chemotherapy and delayed surgery. From 1973--1975, 31 patients with biopsied primary osteogenic sarcoma were treated with preoperative chemotherapy followed by surgical ablation of the primary tumor. Surgery was delayed in order to obtain a custom-fitted prosthetic bone implant in an attempt to avoid amputation. Preoperative chemotherapy included high dose methotrexate (HDMTX) with citrovorum factor rescue (CFR) and adriamycin (T-5 protocol) and was administered for 3 months preoperatively and continued with the inclusion of cyclophosphamide for approximately 5 months postoperatively. At a follow-up period of 30--52 months, 23 of 31 patients (75%) are surviving (21 of 23 with no evidence of disease). Histologic examination of primary tumor removed at surgery revealed varying degrees of tumor destruction (from very little effect to no evidence of viable tumor) attributable to the effect of chemotherapy. The 21 patients that are disease-free survivors had a more complete effect of preoperative chemotherapy on the primary tumor. Some patients achieving favorable effects upon the primary tumor did so only after the dose of HDMTX was escalated to greater than the starting dose of 8 g/m2. Preoperative chemotherapy for all patients with osteogenic sarcoma would seem to offer the following advantages: 1) Evaluation of the effect of HDMTX with CFR on the primary tumor with escalation of the dose of HDMTX until a clinical response is observed, thus defining the dose of HDMTX effective in that patient, to be continued postoperatively as adjuvant therapy; 2) The early use of systemic therapy to eradicate distant microfoci of disease that will eventually kill the patient if not adequately treated by effective chemotherapy; 3) Allow more time for postoperative healing without the need to start adjuvant chemotherapy immediately; and 4) Provide the surgeon time to plan resection surgery. To date, 20 additional patients with biopsy proven osteogenic sarcoma have been treated with more aggressive preoperative chemotherapy (T-7) for approximately 2 1/2 months prior to definitive surgery (resection or amputation). Doses of HDMTX were escalated where necessary and good clinical responses were obtained in 19 of 20 patients. In the majority of patients, no evidence of viable tumor was found on histologic examination of the surgically removed primary tumor. All 20 patients are surviving free of active disease at this brief follow-up period of 4--20 months.


Carcinoma of the esophagus: pretreatment assessment, correlation of radiation treatment parameters with survival, and identification and management of radiation treatment failure. Between January 1969 and February 1975, 344 patients with carcinoma of the esophagus were managed primarily at the Princess Margaret Hospital, Toronto. One hundred sixty-eight (168) of the patients were treated palliatively and 176 of the patients were treated by radical doses of radiation, surgical resection or both. Survival of the radical treatment group was biphasic, the steeper component being identical to the survival of the palliative treatment group, thereby representing a group of patients that did not respond to radical treatment. Analysis of pretreatment assessment parameters indicated that all patients with T1 lesions (length less than or equal to 5 cm, circumference incomplete) and all patients with Stage I disease responded to treatment. Patients who were female, age greater than or equal to 70 years, N0 or had well differentiated squamous cell histology, responded to treatment in at least 80% of cases. No patient with extralymphatic distant metastases responded to treatment. The presence of other major disease did not affect response to treatment. Thirty patients had surgical resections and their survival was not significantly greater than the 146 patients who had radical radiation alone. Survival analysis revealed an optimum range of nominal standard dose (NSD) of 1602--1714 rets (median 1679 rets) for patients treated by radiation alone. An optimum port size (area) of 100--140 cm2 was observed for patients receiving 5000 rads and supervoltage irradiation gave a significantly improved survival in comparison with megavoltage irradiation. Sixty-seven percent (67%) of patients treated by radical doses or radiation developed esophageal strictures postradiation and on the basis of radiological, endoscopic or histological evidence 75% of these strictures were considered to be associated with the persistence of malignancy. On the basis of postmortem examinations (32) and death certificates there was overall an 80% failure to control the disease locally and 95% of strictures were associated with persistence of malignancy in the esophagus. Thirty-one of the 146 patients receiving radical radiation alone had palliation for esophageal obstruction following radiotherapy. The construction of a physiological bypass (e.g., colon) resulted in a mean survival of 215 days which was much longer than the survival observed with rigid esophageal tubes (35 days) or gastrostomy tubes (58 days).


Occurrence of a thermostable antigen of ovarian carcinoma in normal tissues and secretions. Thermostable antigen (TA) occurs in ovarian carcinoma and in certain specialized tissues. High titers of TA were found in nineteen of twenty endocervical extracts but not in myometrium, endometrium, or exocervix. TA was present in all of sixteen cervical mucus samples. Antibodies to perchloric acid extract of bronchus showed cross-reaticity with TA from ovarian carcinoma and cervix. Immunofluorescence with frozen sections revealed TA to be present in the columnar epithelium of ovarian neoplasms, and endocervical glands, in bronchial seromucous glands and in hepatic bile canaliculi. Immunofluorescence and immunodiffusion were used to test for cancer patient antibodies against TA. All sera tested negative. An immunofluorescence inhibition test for circulating TA also tested negative in sera of ovarian and cervical cancer patients.


Induction chemotherapy in advanced squamous head and neck carcinoma with high-dose cis-platinum and bleomycin infusion. Forty patients with advanced head and neck cancer were treated with combined Cis-platinum-Bleomycin chemotherapy. Cis-diammine dichloroplatinum (DDP) 120 mg/m2 iv was given after prehydration, with mannitol diuresis on Day 1. On Day 3, an initial loading dose of Bleomycin 15 mg/m2 was given by rapid iv push followed by continuous 24 hour intravenous infusion of Bleomycin 15 mg/m2 Day 3 through Day 10. DDP 120 mg/m2 iv was administered again on Day 22. The patients were evaluated for tumor response and resectability between Day 29 to Day 35. Of 39 patients who were evaluable, there were 8 complete responses or CR (20%) and 22 partial responses or PR (56%), for a major response rate of 76%. Nineteen patients had surgery (14 patients whose lesions were initially inoperable and 5 patients who were initially operable). Chemotherapy toxicity in 40 patients included alopecia (40), vomiting (39), mucositis (11), skin rash (10), fever (17), weight loss of more than 5 lbs. (25), WBC less than 3,000 (2), platelets less than 100,000 (1), peak serum creatinine of 2 mg% (3), severe-hearing loss (1), hypersensitivity reaction (2). Surgical complication in 19 patients were pharyngocutaneous fistulae (2), wound dehiscence (1), meningitis and brain abscess (1). There was one death secondary to nephrotoxicity. This particular combination chemotherapy when given as initial treatment, appears very effective in reduction of tumor bulk. Long-term follow-up and randomization is necessary to determine effect upon survival.


Chemotherapy of advanced head and neck cancer with methotrexate, bleomycin, and cis-diamminedichloroplatinum II in an effective outpatient schedule. Thirty-one patients with advanced cancer of the head and neck, twenty-six of whom had failed prior irradiation, were treated with an out-patient chemotherapy regimen combining methotrexate, bleomycin, and cis-diamminedichloroplatinum (II). The overall response rate among evaluable patients was 61%, with 22.5% complete remission. If 6 additional patients not completing 3 weeks of treatment are included, the response rate was 51.4%. The median duration of partial remission was 3 months, but none of the complete remitters has relapsed with a follow-up of two to twenty months (median six). Response rate was not dependent on performance status, prior irradiation, or primary site. Toxicity was mild. The remission rate and duration suggest that this regimen may be superior to single agents, and as effective as more complicated and toxic regimens using higher doses of the same drugs.


Evidence for antigenic distinctness of the Epstein--Barr virus-determined nuclear antigen and the Herpesvirus papio-determined nuclear antigen. Previous serological evidence indicated that Epstein--Barr virus-determined nuclear antigen (EBNA) has cross reactive components with the herpesvirus papio-determined nuclear antigen (HUPNA) and, in addition, it has distinct components of its own, not present in HUPNA. In the present study, this hypothesis was studied by absorption experiments. Absorption of anti-EBNA positive human sera with EBNA abolished both anti-EBNA and anti-HUPNA reactivity. Absorption with HUPNA removed all anti-HUPNA reactivity but did not reduce anti-EBNA to any detectable degree, thus confirming the hypothesis.


The effect of combination chemotherapy on neutrophil function in cancer patients. The effect of adjuvant and palliative combination chemotherapy (Vincristine, MeCCNU, 5-Fluorouracil) on neutrophil function was studied in 18 patients prior to and during the anticancer treatment. Neutrophil responsiveness was assessed by leukocyte chemotaxis and chemiluminescence. No impairment could be observed in the circulating phagocyte function in either of the 2 groups before or in connection with short or long-term treatment.