[Chronic infection caused by the simian Mason-Pfizer virus in primate cell cultures]. Cell cultures of monkey prepuce (Rhfs) and African green monkey kidney (BSC-1) were infected once with simian Mason-Pfizer virus (MPV) and virus expression in the course of establishment of chronic infection was studied. The productive infection was characterized by changes in the cell metabolism (DNA synthesis increased 2-3-fold as early as the "zero" passage), the appearance of gs-antigen, formation of virions of type D and high activity of RNA-dependent DNA-polymerase. Multinuclear giant cells appeared only in the infected Rhfs cell culture most sensitive to MPV. In human embryo kidney culture (HEK) productive infection was also established however, HEK cultures did not survive after 3-4 passages. No signs of transformation could be found in any of the cultures studied.


[Reversion in the natural variability of influenza A virus]. In mid-November, 1977, local outbreaks of acute respiratory diseases (ARD) in institutionalized communities began to be recorded in a number of geographical zones of the USSR, and by the end of the month a general increase in the incidence was observed in some areas of the country. The epidemic outbreaks extended gradually and were characterized by moderate development involving mainly young subjects. The strains causing the epidemic had no antigenic relationship with reference A (H3N2) virus and the H1 test were neutralized with antisera to influenza A virus with the antigenic formula H1N1 to 1 1/4 titer. Their neuraminidase was inhibited by the antiserum to the recombinants containing neuraminidase of the subtype N1, especially A/New Jersey 8/76. These results permit to classify the strains causing the epidemic outbreaks in November--December, 1977, as influenza A virus with the antigenic formula H1N1.


[Effective urodynamic evaluation in hospital and general practice (author's transl)]. Urodynamic diagnostic procedure have provided new concepts with consequent advantages for urological patients. The postoperative evaluation of renal pelvic surgery, the detection of functional stenosis of the ureter, stress incontinance, prostatic hypertrophy and postoperative incontinence are typical examples of conditions ideally suited to urodynamic assessment. Urodynamic investigation represents a very important contribution towards the prevention of postoperative failure and enables the achievement of higher diagnostic accuracy.


[Microfluorometric method of determining the quality of reference sera labelled with FITC]. A microfluorimetric method for the determination of the quality of FITC-labeled antisera with the determination of the titer and the optimal working dilution in the FA test is described. Besides, the method permits to determine the confidence factor for any dilution of FITC-antiserum and to compare different lots.


[Study of human diploid cell strains for the presence of oncornavirus determinants]. The WI-38 strain of human diploid cells (HDC) and two strains, L-58 and L-63 isolated in Moscow Research Institute of Virus Preparations were examined in the course of sequential passages for the presence of primate oncornaviruses and their antigens. In parallel, the tumorigenic capacity of the cells was tested by inoculating them into the cheek pouch tissue of Syrian hamsters. In all cases the examined HDC cultures contained no oncornavirus particles or their antigens and in no case showed any infectious activity determined in the KC-test. Only on one subculture of the L-63 strain cells at the 44th passage was a positive result obtained in immunoprecipitation test with antiserum to the LPV oncornavirus. Cells of the L-58 strain which had undergone 27 and 43 passages showed tumorigenicity in hamster experiments. In subsequent passages of these HDC strains no such properties were found. The regular control for the absence of oncogenic message is a necessary condition for the use of HDC strains in virological practice.


[Persistence of the Sindbis virus in cultures producing oncornavirus]. Continuous lines were obtained from primary cultures of BALB/C mouse embryo cells which were found by electron microscope and reverse transcriptase reaction to produce permanently oncoronavirus type C after exogenous infection with Rauscher leukemia virus (RLV). Sindbis virus (SV) was inoculated into virogenic cultures 398 days after infection with RLV. The system in characterized by rapid (3-21 days) disappearance of the infectious arbovirus from the medium and the cells, long-term (over 5 months) persistence on SV noninfectious antigen and signs of stimulation of oncornavirus activity. The level of reverse transcriptase activity in cultures in the presence of persisting arbovirus was 1.5-3.3-fold higher than in cultures infected with RLV alone. Two variants of the course of mixed chronic infection of the cultures with oncornavirus and arbovirus differing in the rate of transition of the arbovirus into the noninfectious form and inhibition or stimulation of oncornavirus functions are discussed.


[Immune ascitic fluids in rats to the Sindbis virus]. Data are presented concerning preparation of universal immune ascitic fluid (IAF) to Sindbis virus possessing a high activity and specificity in different serological tests: CFT, HI, NT, AGDPT, and direct immunofluorescence test. The advantages of rat IAF over the serum of these animals consists in a higher specific activity, lower amount of nonspecific hemagglutination inhibitors and waste protein substances. Rat IAF may be used for production of highly specific diagnostic preparations.


[Use of the immunofluorescence method for the diagnosis of cytomegalovirus infection]. The method of immunofluorescence for diagnosis of cytomegalic infection vitally and in cases of fatal outcomes of the infection is proposed. Recommendations for the preparation of anticytomegalovirus immune sera for an indirect modification of the method are presented alongside with the characteristics of procedure for the immunofluorescent analysis of cytomegalovirus. The clinico-laboratory diagnostic studies have demonstrated certain advantages of the proposed method over virus isolation from patients.


[Method of preparation and evaluation of antisera for the differential determination of secretory immunoglobulin A and free secretory component in biological fluids]. One of the pressing tasks in the study of local nonsusceptibility to infectious diseases and immunochemical analysis of the external secretion is recording of the level of various forms of the secretory IgA (SIgA) and of the secretory component (SC) in various biological fluids. Indication and measurment of the concentrations of the mentioned proteins encounter serious difficulties caused by heterogeneity of their molecular forms. It was shown that the antisera to the whole molecule of SIgA and SC are of no use. On the basis of a new method of purification of free SC and technology of preparation of monospecific antisera capable of separation of SIgA and free SC there were obtained diagnostic antisera for the quantitative recording and differentiation of various forms of IgA and SC in biological fluids. A reliable measurement of the SIgA and SC concentration in some external secretion was carried out with the aid of the mentioned preparations without any complicated chromatographic experiments.


[Effectiveness of the tularemia-antibody erythrocyte diagnostic agent for the detection of specific antigen and antibodies to it]. The authors demonstrated the efficacy of utilization of tularemia antibody erythrocytic diagnostic agent. Data are presented indicating a strict specificity and a high sensitivity of this diagnostic agent for detection of the Vi-antigen both in the tularemia microbe cultures and in the suspensions of the organs of rodents which perished of tularemia, irrespective of the state of cadaver. There was revealed relationship between the sensitivity of the diagnostic agent and the virulence of the tularemia microbe cultures. A possibility of using the diagnostic agent in the antigen neutralization test for detection of specific antibodies in the sera of patients, who sustained the disease, and of the vaccinated humans and animals was revealed.


[Effect of decomplemantation on the course of several allergic skin reactions]. Experiments were conducted on mongrel albino mice; a study was made of the role played by the complement in the development of passive skin anaphylaxis after Ovarie and the so called reverse direct allergic (cytotoxic) reaction. Preliminary animal decomplementation 3 hours before the reactions were conducted failed to alter the intensity of Ovarie reaction, and decreased the intensity of the reverse direct allergic reaction significantly. Reproduction of Ovarie reaction and of the reverse direct allergic reaction on animals 24 hours after the decomplementation did not influence the manifestation of both reactions.


[Reasons for the decrease in viral hepatitis morbidity in Orekhovo-Zueve]. Factors of action on the epidemic process in viral hepatitis in Orekhovo-Zuevo leading to a sharp reduction of the incidence of this disease (which before failed to differ from the mean level in the USSR) -- from 154.0 per 100 thousand residents in 1970 to 36.5 per 100 thousand residents in 1975 are considered from the point of view of discussion of the work of Gromashevsky and Birinboim (ZMEI No. 11, 1976). The decisive significance of measures directed to reduction of the incidence of intestinal infections and gamma-globulin vaccinations is refuted. There was revealed a direct 100% correlation between the incidence of positive benzidine tests for blood contamination of medical instruments and the changes in the viral hepatitis morbidity. Planned measures directed to the elimination of factors of parenteral transmission of the infection carried out intensively from 1970 led to reduction of the incidence of viral hepatitis.


[Immunochemical assessment of the antigenic relationship between the pancreatic kallikrein of rodents and carnivores]. Using quantitative and qualitative immunochemical methods, in combination with isoelectrical focusing, it was shown that antibodies to commercial kallikrein are involved into precipitation reaction with kallikrein in extracts from the pancreas of dogs, cats, guinea pigs, rats and mice. Complexes antigen--antibody exhibit similar physico-chemical properties; during isoelectric focusing, they are found in the same pH range as the immunoactive portion of commercial kallikrein. These data indicate antigenic similarity of pancreatic kallikrein in different mammalian species.


[Flagellar antigens of E. coli serologically interrlelated to H40, 41 and H41, 42, 43 flagellar antigens of Citrobacter. New H-antigeny E. coli i Citrobacter]. The authors confirmed the reference of the test strains H13 (P6c) and H22 (A231a) of the international collection of E. coli to Citrobacter; their antigenic formula was established. As shown, strains P6c possessed a variety of the H-antigen which was not described in Citrobacter earlier, designated as H41a, 97. Three types of flagellar antigens characterized by the presence of an interrelationship with the partial factor H41 of the flagellar Citrobacter antigens were revealed in E. coli; the partial composition of H-antigenic components common for E. coli and Citrobacter was studied. Two of three new varieties of the E. coli H-antigen revealed was characterized by a cross correlation and a relation to the standard H19 E. coli antigen. The strain with the third variety of the H-antigen was capable of forming the H-antigenic mutants which acquired the antigenic component identical to the standard H16 E. coli antigen. E. coli strain is recommended for the replacement of the strain P6c in the International collection of E. coli.


[Determination of the quality of fluorescent antibodies against human globulins using an antinuclear antibody standard]. The authors present data concerning the standardization of the assessment of fluorescent antibodies against human globulins. Fluorescent emision proved to depend on the biochemical indices of the conjugates and on the quality of the initial raw material of crude serum. Materials of quantitative recording of the strength of fluorescence of the nuclear antigen treated with fluorescent antibodies are presented. A difference between the activity and the specific activity (with respect to normal human serum) of the preparations was noted.


Demonstration of RNA-dependent DNA polymerase activity in non-disrupted type D-retravirus particles after treatment with EDTA. The type D-retraviruses PMF virus (PMFV) and Mason-Pfizer moneky virus (MPMV) show RNA-dependent DNA polymerase (revertase) activity after treatment of the nondisrupted virions with the chelating agent ethylenediaminetetraacetate (EDTA). In the range of 1 to 5 mM, the effect is dependent on the concentration of EDTA. As compared with Nonidet P40 disrupted particles treatment of PMFV with EDTA results in a revertase activity of about 65%. Simultaneous addition of Mg- or Ca-ions prevents the EDTA effect. Other divalent cation-binding agents (o-phenathroline, thiosemicarbazide) do not induce an EDTA-like effect. It is suggested that EDTA chelates divalent ions responsible for the structural organization of the viral membrane which, after chelation, gets permeable for the exogenous revertase template.


Immunoglobulin-bearing cells in leprosy. Peripheral blood lymphocytes of 28 untreated and 17 treated patients with different types of leprosy were investigated for the occurrence of immunoglobulin (Ig) bearing cells by means of a smear method. Seven healthy Africans served as controls. In a later stage a complementary study was performed on 6 tuberculoid and 6 lepromatous leprosy patients by means of a suspension method. The immunofluorescence technique was used for the detection of Ig-bearing cells, In tuberculoid leprosy an increase of Ig-bearing cells seems to occur during treatment, predominantly expressed by an increase in IgD-bearing cells. In lepromatous leprosy no increased percentages of Ig-bearing cells were observed.


Syringocystadenoma papilliferum: light and electron microscopic studies. Light microscopic and electron microscopic studies of a lesion displaying a typical histopathological picture of syringocystadenoma papilliferum (S.P.) are reported. Light microscopic serial sections showed that the tumor parenchyma extended to the epithelium just above the intrafollicular sebaceous duct. The tumor thus presumably develops from the epithelial region corresponding to the intrafollicular duct of the apocrine sweat gland. Electron microscopy revealed a non-keratinized intracytoplasmic cavity and intercellular canaliculi apparently formed as a continuation of this cavity. Keratinized cells were absent from the glandular and duct epithelium and even from the superficial epithelial portion of acanthotic areas. Neither myoepithelial cells nor secretory granules were identified in areas showing tubular, glandular, or sinusoidal structures. It was concluded that the tumor differentiates towards both the intrafollicular and intradermal duct of the embryonic apocrine sweat gland apparatus.


Antenatal assessment of fetal age and maturity using fetal fat staining cells in amniotic fluid. A cross sectional retrospective study of 400 cases of amniocentesis was performed to evaluate the reliability of using fetal fat staining cells as a method for assessment of fetal age and maturity. A sharp rise in the percentage of these cells in amniotic fluid occurred after 37 weeks of gestation and a level of 20 per cent or more indicated fetal age of 38 weeks or more, corresponding to maturity by our definition. The false positive and negative rates were 3.5 per cent and 14 per cent respectively. The pre-eclamptics, diabetics and Rh sensitized groups fell within the distribution of the normal population. The dysmature group, however, showed an earlier rise (20% at 36 weeks). The origin of the fetal fat staining cells and the reasons for the early rise in dysmaturity are discussed. Estimation of fetal fat staining cells in the amniotic fluids using the 20 per cent level is a reliable test for maturity in normal and some abnormal pregnancies, excluding dysmaturity.


The inverted follicular keratosis. A surprising variant of seborrheic wart. The pathological features of 29 lesions that had been diagnosed as inverted follicular keratoma have been reviewed. The clinical data suggest that these lesions occur predominantly on the face of middle-aged or elderly man. None were correctly diagnosed before removal. The main histological points noted were the presence of an invaginated type of architecture, foci of keratinization (squamous eddies) and areas of basaloid cells which were arranged haphazardly with mitotic figures (Borst-Jadassohn change). All the histological features seen are also observed in irritated or traumatized seborrheic warts and no evidence was found for the original suggestion that these lesions derive from hair follicles.


Clinical and histopathological studies on spotted grouped pigmented nevi with special reference to eccrine-centered nevus. Clinical and histopathological studies on 16 cases of spotted grouped pigmented nevus resulted in this disease being classified into at least three types. Characteristic findings of each type are described in this paper, in particular that nevus cell proliferation, a finding common to each type, is closely related pathogenetically to skin appendages, especially with eccrine sweat ducts. The eccrine-centered nevus characterized by Mishima corresponds to the third type, according to the present author's classification. The similar characteristic histopathological finding was also observed in a specimen taken from a black papule in a case of giant pigmented nevus with disseminated pigment cell nevi. We would like to consider that the cells of eccrine-centered nevus are derived from nevoblasts present in the walls of sweat ducts.


Hairy cell leukemia (leukemic reticuloendotheliosis) in serous effusions. The cytology of two cases of hairy cell leukemia (leukemic reticuloendotheliosis) involving pleural and ascitic fluids is described. With the Papanicolaou stain, the cells have a "lymphoid" appearance. The presence of these cells plus knowledge of the clinical history might help the cytologist and/or hematologist in their identification. A tartrate resistant acid phosphatase (TRAP) stain will confirm the true nature of such cells and establish the clinical diagnosis of hairy cell leukemia. Unlike the other members of the leukemia-malignant lymphoma group, the body cavities are rarely involved.


Mechanism of recurrence of pigmented nevi following dermabrasion. A study was made of pigment freckles which recur after skin abrasion of spotted grouped pigmented nevi, according to the lapse of time, chiefly by means of the fluorescence method of Falck and Hillarp. The mechanism of recurrence is summarized as follows. Nearly simultaneously with the epidermal regeneration, dendritic melanin-producing cells derived from hair follicles and eccrine sweat ducts appear in the basal and prickle-cell layers of epidermis. These cells then create junction activity in the basal layer of epidermis, in the hair follicles and in the eccrine sweat duct walls. Finally, these nevus cells drop off into the underlying layer of scar tissue along the epidermal appendages. The dendritic melanin-producing cells seen in the early regenerative pigment freckles were thought to be incompletely differentiated nevus cells in the pre-stage of junction nevus formation. Appearance of dendritic melanin-producing cells and formation of junction activity in the eccrine sweat duct walls suggest the following possibilities. (1) In the cases of spotted grouped pigmented nevus which we studied, nevus cell proliferation also occurred eccrine-centrically on pathogenesis. (2) An important role is played by the eccrine sweat ducts in the recurrence of ordinary pigmented nevi after incomplete removal. (3) Eccrine-centered nevus cells are derived from nevoblasts in the eccrine sweat duct walls.


Identification of mononuclear cells infiltrating basal cell carcinomas. The identification of mononuclear cells extracted from 12 basal cell carcinomas has been carried out by means of various assays such as the E rosette test, FITC anti-Ig conjugates and peroxidase conjugates viewed at an ultrastructural level. The relative quantities of B and T cells have been determined as well as the morphology of the extracted cells. It was found that the T/B ratio is close to that of a delayed-type reaction to tuberculin, suggesting a possible identical immune mechanism, and also that the morphological appearance of some tissue lymphocytes differs from that of circulating lymphocytes due to the presence of both membrane and cytoplasmic Ig.


The fate of tetrachlorosalicylanilide in photosensitized guinea pigs. The distribution and fate of 3, 3', 4', 5-tetrachlorosalicylanilide (TCSA) in photosensitized guinea-pig skin following topical application at varying times was investigated by fluorescence microscopy in unfixed, frozen sections. When the site to which TCSA had been applied was irradiated with long-wave UV light, TCSA was detected in the entire epidermis only 3 days after application and in the horny layer up to 10 days. Unless TCSA-treated sites were exposed to light, TCSA was still recognized in the entire epidermis 10 days after application and in the horny layer even at 3 weeks. The irradiated photosensitized guinea pigs in the present study eliminated TCSA more rapidly than normal control animals in our previous work. The mechanism of persistent light reaction was discussed and we concluded that TCSA can remain in the skin for a long time, because it is a prohapten, not a hapten.


Phototoxic reaction to chlorpromazine as studied with the quantitative mouse tail technique. A quantitative in vivo technique for the study of the phototoxic reaction to systemic chlorpromazine has been developed, based on the increasing tissue fluid content of the albino mouse tail. A maximum increase in tissue fluid of about 25% was registered 24 hours after starting the long-wave ultraviolet light exposure. The minimum phototoxic dose of chlorpromazine was 2.5 mg/kg.


The leukocyte migration test in nickel contact dermatitis. Ten patients with contact dermatitis to nickel, confirmed by patch testing, were investigated by the leukocyte migration test. Specific inhibition of leukocyte migration was obtained with nickel protein complexes. Bovine albumin, human albumin and human epidermis protein were used as carrier proteins in the complexes. Specific inhibition of leukocytes from patients with positive patch test to nickel could not be demonstrated with the hapten alone.


The nitroblue tetrazolium test in acne vulgaris. The nitroblue tetrazolium (N.B.T.) test has been studied in 19 patients with moderate or severe acne vulgaris and 18 patients with subclinical or no acne. There was a significant elevation of the N.B.T. score in those patients with moderate or severe acne as compared with the control group. This is probably a result of the inflammatory reaction in and around the involved sebaceous gland follicles.


Treatment of psoriasis with trioxsalen baths and dysprosium lamps. Photochemotherapeutic treatment of psoriasis with trioxsalen baths (0.5 mg/1) for 15 minutes followed by irradiation with dysprosium lamps (Osram HQI-TS) healed or nearly healed the psoriatic lesions in 18 patients within 3-5 weeks. A control area treated with the Ingram method showed a slower healing in 9 of these patients. Methoxsalen bath was not as effective in healing at the concentration used (1 mg/l). The bath method is easy to administer and cosmetically acceptable. Sensitisation to light is maximal immediately after the bath and disappears more quickly than after painting with an alcoholic trioxsalen solution. By using baths, there is less risk of accidental burns or uneven pigmentation than with the often time-consuming local application of psoralen solutions. Toxic systemic effects, which are possible with oral treatment, are less apt to occur. The dysprosium lamps give high intensity in the UV-A region. Exposure times of 10 seconds to 8 minutes are effective in the treatment of psoriasis, where both the UV-B region itself and the UV-A in combination with trioxsalen have psoriasis-healing properties.


Necrolytic migratory erythema and glucagon cell tumour of the pancreas: the glucagonoma syndrome. Report of two cases. Two cases of necrolytic migratory erythema are described. Both patients also suffered from anaemia, weight loss, hypersedimentation and carbohydrate intolerance. A solitary pancreatic tumour was found in both cases-- at autopsy in one and at laparotomy in the other. Microscopic examination of skin biopsies showed necrolysis of superficial epidermis. Both patients had extremely elevated plasma concentrations of pancreatic glucagon. By means of specific staining and immunofluorescence techniques the tumours were shown to consist of glucagon-containing alpha2-cells. It is concluded that these patients suffered from the newly described glucagonoma syndrome.


Syndermatotic cataract in patients with psoriasis. 266 consecutive in-patients, more than 10 years of age (mean age 24.7) and suffering from psoriasis have been examined with slit lamp and ophthalmoscope to determine the incidence of cataract among such patients. 188 (70%) had clear lenses. 66(26%) presented some minute punctate opacities considered as physiological variations. Four had blue-dot cataract, and 6 had congenital or evolutionary small opacities at various sites in the lenses but without blurring of vision. Only one patient, aged 72, had crystalline cataract with slightly impaired vision. The conclusion is drawn that the incidence of cataract among patients with psoriasis does not exceed that in the normal population. Consequently, routine eye examinations for cataract are unnecessary in these patients.


Dermatitis herpetiformis in association with treated coeliac disease. The paper describes an adult patient with coeliac disease who developed classical dermatitis herpetiformis while receiving a gluten-free diet. The dermatitis is controlled by sulphapyridine but relapses when this is withdrawn, even though the patient continues with the diet, indicating that a gluten-free diet plays little or no part in controlling the skin condition.


Recurrent bullous eruptions on the lower legs. In 15 patients with venous or arterial insufficiency of the legs, bullous eruptions were observed on the lower legs. The bullae were subepidermally situated. Circulating antibodies against the basement membrane zone were not demonstrated in 2 cases investigated. This eruption seems to represent a clinical entity of unknown etiology.


Metastasizing erythroplasia Queyrat. Report of a case. A typical clinical and histologic case of Queyrat's erythroplasia of the glans penis is presented. For several years the patient had been treated for balanitis and inflammation of the glans. A few months before the patient died, the correct diagnosis was established. Shortly after, metastases to the inguinal lymph nodes were found and treated with X-rays. At autopsy, metastases to other organs were revealed. It is concluded that erythroplasia of Queyrat might be Bowen's disease of a mucosal or mucocutaneous area, and this "carcinoma in situ" may metastasize. Consequently Queyrat's erythroplasia should be treated as a malignant disorder as soon as the diagnosis is established.


Determination of cyclic AMP in heat-separated human epidermal tissue. Heat separation is introduced as a simple, reliable and quick method for obtaining pure epidermis for cyclic AMP analysis. Heating at 60 degrees -65degrees C for 10 min resulted in a distinct separation of epidermis from dermis. The level of cyclic AMP in epidermal tissue from 9 patients was 3.93 +/- 0.31 pmol per mg dry weight. The inter- and intra-assay variations were 10.3% and 8.8%, respectively. The coefficience of variation in eight biopsies from the same piece of skin was 16.2% The sensitivity of the assay was 0.07 pmol. Recovery analyses revealed that 80.5% of labelled cyclic AMP was refound after extraction and purification procedures. Addition of unlabelled cyclic AMP resulted in a recovery of approximately 100%. The specificity of the assay was high without interference from several nucleotides and the validity was confirmed by zero values obtained after addition of phosphodiesterase or charcoal.


Enzymatic liberation of viable cells of human skin. To obtain viable cells from normal human skin, clostridial collagenase was used. Crude collagenase digestion of collagen fibres and basal lamina results in free dermal cells and sheets of epidermis. The collagenase was tested at various concentrations, solvents and incubation periods. The specimens digested were either split or full thickness skin of varying size. The optimal result was obtained by using small (3mm across) split skin pieces incubated in 2 mg/ml collagenase. The choice of solvent MEM, MEM supplemented with serum, and Tris buffer, was less important. 3 hours' incubation the epidermis was peeled off in sheets and finally dissociated by trypsin-EDTA. The corium was completely digested after 6 hours. After 6 hours' incubation no viable cells could be seen. The epidermal cells appeared mainly as polygonal cells of various sizes and a few little dendritic cells. The dermal cells had a heterogeneous morphology during the first weeks of cultivation. After 2 weeks the cells appeared as fibroblast-like cells.


The cytolytic effect of PUVA treatment on PHA-stimulated human peripheral lymphocytes. The morphology of PUVA-treated and PHA-stimulated human peripheral lymphocytes in vitro was studied by means of the light and electron microscopes. Most cells showed signs of cytolysis, such as pyknotic nuclei and a swollen and decomposed cytoplasm; a few cells, however, appeared as normal lymphoblasts. In the latter, repair-enzyme mechanisms may successfully have rectified alterations in the DNS prior to the inception of S-phase. These findings are discussed with reference to the in vivo effect of PUVA treatment.


Prostaglandins in PUVA-treated psoriasis. The biosynthesis of prostaglandins (PG) in biopsies from 9 patients with recalcitrant psoriasis was studied before, during, and after treatment with 8-methoxy-psoralen and long-wave ultraviolet light (UVA). No statistical difference was found between the results obtained before, during, and after the treatment. In PGE1-equivalents, the concentration in involved psoriatic skin was 2.16 +/- 0.31 ng (mean +/- S.E.M.) before, 2.01 +/- 0.33 ng during, and 2.85 +/- 0.31 ng per g wet weight after PUVA. In uninvolved skin the concentrations were 2.38 +/- 0.26 ng, 2.23 +/- 0.15 ng and 3.29 +/- 0.24 ng per g wet weight, respectively. In the presence of exogenous arachidonic acid in the incubation medium the activity formed was higher, but no statistical difference was found between pretreatment values and those obtained during and after PUVA treatment. The hypothesis that PUVA treatment stimulates PG biosynthesis, thus accounting for the beneficial therapeutic antipsoriatic effect, could not be confirmed.


Melanocyte mitosis in UVB-irradiated mouse skin. Little is known about the mitotic activity of the epidermal melanocyte population. Vincristine sulphate has now been used in vivo to study the frequency of mitosis of epedermal melanocytes in the ear skin of C57BL mice. It is shown that there is continuous formation of new melanocytes in unstimulated skin due to mitosis of pigment-producing epidermal melanocytes. The increase in this mitotic rate following daily UVB irradiation suggests that this is a major mechanism responsible for the UV-induced increase in the epidermal melanocyte population.


Ultrastructure of giant pigment granules in lentigo simplex. Electron microscopic observation of lentigo simplex on the sole revealed the presence in the lesion of giant pigment granules in melanocytes and keratinocytes. The giant granules were membrane-bound bodies containing electron-dense amorphous substances and less electron-dense microvesicles. It was also revealed that compound melanosomes, similar in size and shape to giant pigment granules, were present in melanocytes. Within the compound melanosomes, melanosomes showed disintergration into electron-dense fine particles concomitant with the release of less electron-dense microvesicles. These fine particles then aggregated to the mass of electrondense amorphous substances  which eventually embedded the microvesicles. This evidence strongly suggests that the giant pigment granules are formed by complete degradation of melanosomes in the large compound melanosomes which can arise within melanocytes by autophagy.


Pigmented nevus divided between recipient and donor sites by intermediate skin grafting procedure: a study with fluorescence method (Falck & Hillarp). Pigment freckles seen in donor and recipient sites at intermediate skin grafting were studied, chiefly by means of Falck & Hillarp's fluorescence method, with the following results. (1) An important role is played by the regenerative eccrine sweat ducts in the mechanism of recurrence of so-called "lentigines" after incomplete removal. (2) Junctional activity, not associated with the regeneration of epidermal appendages, can recommence in the epidermis of intradermal nevi. (3) B-type nevus cells once again actively produce melanin. (4) Specific fluorescence was observed corresponding to the melanin granules distributed in the manner of "nuclear caps" in keratinocytes around junctional activities. This latter finding may indicate that melanosomes which still contain DOPA are transferred from melanin-producing cells to keratinocytes.


Pemphigus acantholysis in tissue culture: studies on photo-induction. It has previously been shown tha UV irradiation can give rise to fresh skin lesions in the uninvolved skin of patients with pemphigus. In this study no increase of acantholytic activity was found in an organ culture model system for pemphigus in vitro, after in vitro and in vivo irradiation under various experimental conditions. By immunofluorescence no increase in IgG binding in irradiated specimens was observed. From these in vitro findings, which contrast the effect of UV light in pemphigus in vivo, it may be concluded that the circulatory system of the skin plays a major role in the photo induction of acantholysis in pemphigus.


Epidermolysis bullosa acquisita and Crohn's disease. A patient with epidermolysis bullosa acquisita (EBA) associated with Crohn's disease is presented. The clinical, histological and immunological findings were in keeping with previous reports. However, clinically normal skin and mucosa exhibited deposits of IgG and C3 in the basement-membrane zone. These deposits remained unchanged during the treatment period. It is therefore suggested that immunological mechanisms are implicated in pathogenesis of the disease.


Lymphocyte transformation induced by nickel sulphate: an in vitro study of subjects with and without a positive nickel patch test. Lymphocytes from 8 patients with contact dermatitis and a positive nickel patch test, 7 patients with contact dermatitis due to other factors and with a negative nickel patch test, and 9 other subjects, 7 of whom suffered from other dermatological disorders, were tested with the lymphocyte transformation test (LTT), using nickel sulphate in various concentrations. All execpt one of the nickel allergics showed a significant response to nickel, whereas the controls showed borderline (3 patients) or no response (12 patients). These observations confirm previous findings indicating that nickel hypersensitivity can be diagnosed in vitro. This may be of importance in cases of acute contact dermatitis where patch testing is undesirable. Some individuals with a negative nickel patch test responded significantly though weakly to nickel in the LTT, indicating that apart from acting as a specific antigen (hapten?) nickel sulphate may also have weak non-specific mitogenic properties.


Local absorption of zinc from wounds treated with various zinc-compounds. It is shown that in the rat, zinc is absorbed from excisional wounds into serum, from various locally applied zinc oxide preparations as well as from zinc peroxide and zinc sulphate. The most pronounced absorption of zinc to serum was recorded from wounds treated with zinc peroxide. Microscopically the cellular inflammatory response observed in wounds treated with a gauze sponge impregnated with a zinc sulphate solution was weaker than that found with the other treatment methods used.


Generalized pustular toxic erythema: pathogenetic relationship between pustule and epidermal appendage (hair follicle or sweat duct). Seven patients are described, who had generalized toxic erythema with sterile pustules. Study of serial sections of the pustules confirmed a specific localization to hair follicle or epidermal sweat duct. Five patients had a past history of medications, exposure to an organic solvent, or infections.


Circulating immune complexes in syphilis. 13 patients with syphilis were investigated regarding the presence of circulating immune complexes by the methods of C 1q-binding-activity and anticomplementarity. Elevated C1q-binding-activity was demonstrated in 6 of 7 patients with secondary syphilis, a significantly greater incidence than among patients with primary syphilis and neurosyphilis. Anticomplementarity was demonstrated in five of seven patients with secondary syphilis and in two patients with neurosyphilis. Anticomplementarity was found in only one of four patients with primary syphilis. The presence of immune complexes may be of importance in the aetiology of some of the lesions of secondary and perhaps tertiary syphilis.


5-S-cysteinyldopa and trichochromes in red feathers. Red cock feathers (Rhode Island) were found to contain dopa and cysteinyldopa, trichochromes B and C, and two unidentified trichochromes. Trichochromes E and F were not found. Previous findings of trichochromes E and F may be explained as artifacts.


Free and bound 5-S-cysteinyldopa and dopa in human malignant melanomas. Free dopa and 5-S-cysteinyldopa were extracted from two human melanomas. Subsequent hydrolysis of carefully washed melanoma tissue released dopa and 5-S-cysteinyldopa, indication the presence of these catechol amino acids in proteins. Cysteinyldopa-containing proteins may represent the antigens previously demonstrated in human melanomas.


Deposits of complement and immunoglobulins in dermal and synovial vessels in psoriasis. Deposits of complement C3 and/or immunoglobulin were found in the vessel walls and/or at the dermal-epidermal junction in skin lesions of all of 11 patients with psoriatic arthritis and guttate psoriasis. Similar deposits were seen in 6 out of 15 patients with psoriasis vulgaris. Synovial tissue available from 2 patients revealed deposits in the vessel walls.


Local excision in the treatment of oral discoid lupus erythematosus. Oral lesions in eight patients with discoid lupus erythematosus (DLE)  were treated with local excision. Ten well-demarcated oral lesions were excised and a primary closure was obtained following the excision. Six patients out of seven became symptom-free following the treatment. The median observation time was 1.0 year. The follow-up examination showed erythema in one case and recurrence in one case at the operation site. In the remaining eight operation sites, a soft scar with no sign of recurrence was seen. These results indicate that the method should be investigated further in order to obtain long-term results. Local excision of oral discoid lesions may be a valuable therapeutic supplement in some cases of DLE.


Atypical necrobiosis lipoidica of the face. A 36-year-old healthy man with atypical necrobiosis lipoidica of the face is described. The lesions were annular with a raised erythematous palpable border. The centres were slightly depigmented and atrophic, without telangiectasia or hair loss. Histopathological changes revealed prominent giant cells in small groups without clear granuloma formation. They were located at all levels of the drmis between the collagen bundles.


Pemphigus vulgaris in a 15-year-old girl. Pemphigus vulgaris is uncommon in adolescence and only ten well documented cases in this age group were found in a recent review (5). Because the condition is often more severe in the younger age group it is important to consider it in the differential diagnosis of bullous eruptions of childhood and to perform direct and indirect immunofluorescence studies.


Palpebral cellulitis.

Palpebral cellulitis. Patients with palpebral cellulitis may turn in the first place to a skin clinic. The clinical picture and the course of disease are illustrated here by four case histories. The importance of differentiating between collateral orbital edema, palpebral cellulitis and orbital cellulitis is stressed.


Thyroid function during pregnancy with special reference to hydatidiform mole and hyperemesis. The thyroid function tests, serum protein-bound iodine (PBI), serum triiodothyronine reaction (T3-test) and serum cholesterol were measured in three groups of pregnant women: 1) 57 with normal pregnancy, 2) 35 with hyperemesis and 3) 14 with hydatidiform mole. A pattern of high values of PBI and T3-test and relatively low values of cholesterol in most of the patients with hydatidiform mole, and in almost one third of the patients with hyperemesis was found. A positive correlation of the volume of molar tissue and the values of PBI was observed. The treatment of patients with hyperemesis mostly resulted in normalisation of the thyroid function tests, while this was not the case in patients with mole. It is suggested that there might be a common cause of the thyroid stimulation in patients with mole and hyperemesis.


alpha-Fetoprotein content in liver from rats following hepatotoxin administration and partial hepatectomy. alpha-Fetoprotein (AFP) contents in liver from hepatotoxin-treated or partially hepatectomized rats were quantitatively measured using a radioimmunoassay by freezing and thawing perfused-liver homogenate in Triton X-100 No significantly increased contents of AFP in injured or regenerating livers were found even with a marked increase in serum AFP concentrations of the treated rats, suggesting that AFP could be rapidly secreted into the circulation identically under the entirely different conditions. Increased contents of AFP in liver could be detected only in newborn rats with serum AFP concentrations of more than 10 microgram/ml.


A case of whipple's disease with repeated operations for ileus and complete cure. The authors make a case report of a male patient, 58 of age, whose complaints started 37 years ago; inspite of an operation, repeated medical check-ups and treatment, they had no other choice but to perform a further laparotomy when the patient presented himself with progressive cachexia and signs of an acute abdomen. Taking the history, the operative and histological findings of the patient into consideration, the correct diagnosis could be made: Whipple's disease. The presence of this condition was also proven by postoperative X-rays and immunological examinations. The patient has been restored to health after a year's antibiotic treatment. An impaired defensive ability, mainly of the immune and macrophage system can be proven to exist in Whipple's disease. Two main tasks are essential: 1) to hold the patient under constant control and 2) to continue research with the aim of finding ways of immunocorrection.


Changes in fetal supraventricular extrasystoles during uterine contractions in labour. The electrocardiogram and phonocardiogram were recorded from a fetus exhibiting supraventricular ectopic beats during labour. The ectopic beats showed increased amplitude of the first heart sound (S1) compared to sinus and postectopic beats; the R-S1 interval was prolonged and the mechanical systole, measured as the S1-S2 interval, shortened. During uterine contractions these parameters were measured every 10 sec. The S1 amplitude of the ectopic beats decreased; simultaneously the R-S1 interval increased and the S1-S2 interval shortened. The changes were delayed compared to the amniotic pressure curve. The changes found can be explained by a shift of blood between the fetus and placenta, caused by uterine contraction.


Pseudo-exfoliation fibrils examined by negative staining. Pseudo-exfoliation (PE) material, collected from lenses extracted because of cataract, has been examined by negative staining. The material was fragmented by ultrasound into single fibrils and small collections of fibrils. The fibrils were composed of a small number of filamentous subunits associated with or surrounded by a fuzzy material which had side excrescences at regular intervals of 50--55 nm or 25--30 nm. The side excrescences correspond to the cross-bands of the PE fibrils. It is proposed that the PE fibrils are composed of a few subunits of a fibrillar protein, which form a core to which glycosaminoglycan side chains are attached. The visible periodicity of the PE fibrils, appearing as cross-bands, is thought to be caused by clusters of glycosaminoglycan distributed at regular intervals along the fibrils.


Synthetic disaccharide-protein antigen for production of specific O2 antiserum for immunofluorescence diagnosis of salmonella. Antisera from rabbits immunized with the synthetic disaccharide paratose 1 leads to alpha 3 mannose, representive of Salmonella O-antigen 2, covalently linked to bovine serum albumin (BSA), were used in indirect immunofluorescence studies for the identification of Salmonella serogroup A (O-antigen 1,2,12) bacteria. Among 1311 enteric bacteria tested, 497 were Salmonella. The anti-paratose 1 leads to alpha 3 mannose-BSA serum identified correctly all the 63 serogroup A strains tested. No positive reactions were recorded among 1248 strains respresenting Salmonella other than serogroup A, E. coli, Shigella, Citrobacter, Klebsiella, Enterobacter, Serratia, Proteus, Pseudomonas, Acinetobacter, Vibrio, Yersinia and Bacteroides. The study illustrates the high specificity of the antiserum elicited by immunization with the synthetic disaccharide-protein immunogen.


Plasma proteins in the period of posttransfusion polycythaemia in rats. Plasma proteins in the period of post-transfusion polycythaemia in rats. Acta Physiol. Pol., 1978, 29 (1): 47-54. The plasma protein of polycythaemic rats with strongly inhibited erythropoiesis and of normal rats was characterized with the use of ion exchanger chromatography on DEAE-cellulose in an NaCl concentration gradient and electrophoresis on polyacrylamide gel. Experimental posttransfusion polycythaemia was found to cause considerable changes in the amount and composition of plasma proteins. The amount of gamma and alpha1 globulins increased in polycythaemic animals, whereas albumin content decreased. There also appeared wide differences in the elution profiles obtained from ion exchanger chromatography of both kinds of plasma. Electrophoretic analysis of fractions obtained by chromatography demonstrated a similarity only in the composition of the globulin fractions of both the plasma kinds. The remaining fractions differed widely in their protein composition.


Local effect of single stranded polyadenylic acid on passive cutaneous anaphylaxis in mice. Single stranded polyadenylic acid (Poly A) administered locally inhibited passive cutaneous anaphylaxis in mice. In experiments performed by equilibrium dialysis Poly A was able to bind histamine. The association constant of the reaction was determined, Ka = 1.3 +/- 0.2 x 10(5) I/M. One Poly A molecule can bind maximally two molecules of histamine dichloride. Poly A inhibited the antigen-induced release of histamine from the peritoneal rat mast cells when it was given together with sensitizing antibodies.


The effect of sodium 5,6-dimethyl-2-nitroindanedione on anaphylactic reactions in vitro. A nitroindanedione (BRL 10833) inhibited the antigen induced release of histamine and slow reacting substance of anaphylaxis (SRS-A) from passively sensitized human lung at similar concentrations to those required for the inhibition of histamine release by disodium cromoglycate (DSCG). BRL 10833 was more potent than DSCG as an inhibitor of histamine release by antigen from actively and passively sensitized rat peritoneal cells and rat skin fragments.


Epidemic kepone poisoning in chemical workers. From March 1974 through July 1975, 76 (56%) of 133 persons who had worked at a pesticide plant that produced Kepone, a chlorinated hydrocarbon insecticide, contracted a previously unrecognized clinical illness characterized by nervousness, tremor, weight loss, opsoclonus, pleuritic and joint pain, and oligospermia. Illness incidence rates for production workers (64%) were significantly higher than for nonproduction personnel (16%). The mean blood Kepone level for workers with illness was 2.53 ppm and for those without disease 0.60 ppm (p less than 0.001). Blood Kepone levels in current workers (mean, 3.12 ppm) were higher than those in former employees (1.22 ppm). Blood Kepone levels for workers in nearby businesses and for residents of a community within 1.6 km of the plant ranged from undetectable to 32.5 ppb. Illness attributable to Kepone was found in two wives of Kepone workers; there was no apparent association between frequency of symptoms and proximity to the plant in the survey of the community population.


Two pregnancy-associated serum proteins with pregnancy-specific beta1-glycoprotein determinants. A protein showing strong antigenic cross-reactivity with the "pregnancy-specific" beta1-glycoprotein (PSbetaG) was demonstrated in serum of pregnant women during the second and third trimesters, but not in sera from normal men and nonpregnant women. The described protein had alpha2-electrophoretic mobility and its molecular weight was estimated to be around 200,000 as compared to 80,000 for PSbetaG. Crossed immunoelectrophoretic analysis, using antiserum to PSbetaG, indicated the presence of additional antigenic determinants in PSbetaG not demonstrable in the cross-reacting alpha2-protein. The PSbetaG/alpha2-protein ratio was rather constant in consecutive serum samples from one woman during pregnancy, but showed a marked variation when sera from different women were compared. Results of rocket immunoelectrophoresis gave a rough estimate of the ratio between these two proteins in a sample of pregnancy serum. The reported findings point to the need for development of specific methods for quantitation of the two proteins and the necessity to re-evaluate earlier quantitative PSbetaG data.


Adriamycin alone or in combination in 100 patients with carcinoma of the cervix or vagina. One hundred consecutive evaluable patients with advanced carcinoma of the cervix or vagina were treated with doxorubicin (Adriamycin) alone or in combination. The use of Adriamycin in conjunction with cyclophosphamide and 5-fluorouracil resulted in the highest response rate with all responders continuing to respond.


Improved surgical palliation of advanced carcinoma of the esophagus. Fifty-four patients with far-advanced carcinoma of the esophagus were operated on between the years 1974 and 1976. No attempts were made to resect the lesion. The stomach was used fifty-three times to bypass the lesion and the right colon was used once. In twenty-eight patients the stomach was placed substernally and the anastomosis was done in the neck. Twenty-five patients had the anastomosis to the esophagus done in the chest. The thirty day operative mortality was 7.4 per cent and the average survival was five months. These figures compared favorably with a group of thirty-five patients with far-advanced carcinoma of the esophagus seen between the years 1971 and 1973 and handled with a variety of modalities. In this group the thirty day mortality was 31.4 per cent (11/35) and the average survival was three and a half months.


Hyper-alpha-2-macroglobulinemia in narcotic addicts. Significant increases of serum alpha-2-macroglobulin (alpha-2-M) were detected in narcotic addicts presenting at a methadone treatment center. The mean alpha-2-M level was 341 +/- 14 mg/dl compared with 231 +/- 8 mg/dl in normal persons (p less than 0.01). In a comparable group of alcoholics with laboratory evidence of hepatic dysfunction the mean alpha-2-M level was 208 +/- 10 mg/dl. Although the serum immunoglobulin M content was also substantially elevated in the narcotic-addict group, no correlation was obtained between the serum content of this protein and alpha-2-M. Similarly, no correlation between alpha-2-M level and serum zinc content was observed. When the values of the trypsin-binding activity of serum measured in 13 addicts, 15 alcoholics with laboratory evidence of hepatic dysfunction, and 16 normal subjects were plotted against the amount of alpha-2-M measured in the same subjects, a linear correlation was obtained between trypsin-binding activity and alpha-2-M. Thus, the significantly increased serum trypsin-binding activity observed in the addicts is that which might be expected if normal alpha-2-M is being accumulated in large amounts as a result of increased macroglobulin synthesis.


Lymph-node bacilliform bodies resembling those of Whipple's disease in a patient without intestinal involvement. A 38-year-old man developed symptoms of arthalgias and arthritis, lymphadenopathy, and weight loss. An axillary lymph-node biopsy was done in the diagnostic study; a periodic acid Schiff stain, done for evidence of fungal infection, showed periodic acid Schiff reagent-positive macrophages. Electron microscopy showed the typical morphologic features of the bacilliform bodies associated with Whipple's disease to be present in the macrophages of the lymph node. The patient had no intestinal symptoms. The absorption of a variety of substrates was found to be normal. Nine intestinal biopsies showed no organisms similar to those found in his lymph node. On tetracycline therapy, he symptomatically improved. The findings raise the question of the route of infection in Whipple's disease and point up the usefulness of periodic acid Schiff staining of lymph-node biopsies.


The distribution of myelin basic protein in central nervous system lesions of multiple sclerosis and acute experimental allergic encephalomyelitis. To obtain more information about early events in central nervous system myelin injury in multiple sclerosis (MS), a comparative study was conducted of the distribution of myelin basic protein (BP) in tissue lesions of both MS and experimental allergic encephalomyelitis (EAE). Sixteen postmortem specimens containing lesions from 4 persons with MS and the brains of 14 guinea pigs with acute EAE induced with BP were studied. Cryostat sections of quick-frozen material were fixed, treated with rabbit antibody to BP, and processed by immunoperoxidase techniques. The reaction of antibody to BP was reduced or absent in MS lesions but normal in uninvolved surrounding tissue. Alterations in BP generally paralleled changes in staining of myelin by histological methods. Marked diminution of reactivity with anti-BP occurred in early lesions of MS and extended far beyond any identifiable inflammatory elements. In both parenchyma and perivascular areas, lipid-laden macrophages in MS plaques frequently contained BP material in addition to a variable amount of endogenous peroxidatic activity. Compared with MS, BP was relatively well preserved in brains of guinea pigs with EAE. At the light microscopy level, normal-appearing patterns of BP existed adjacent to the perivascular cellular infiltrates, and macrophages containing BP material were rare. The results of this study suggest differences between MS and acute EAE in both removal of BP from the central nervous system and its subsequent disposal.


Electron microscope visualization of chromatin and other DNA-protein complexes. In this review we have tried to cover the direct mounting techniques currently in use and, through micrographs kindly supplied by our colleagues, to illustrate uses of the different methods. For the novice, it is important to select one technique, taking time to become acquainted with its use in visualizing simple test samples before applying it to examine more complex structures. We have tried to emphasize the importance of controlling the fixation for two reasons. First, confidence in the fixation step allows one to correlate structures observed by EM with structures present in solution. Second, fixation of otherwise labile samples allows one to apply powerful purification techniques.


Bullous pemphigoid and ovarian cystadenocarcinoma. Immunologic studies. A patient with bullous pemphigoid and papillary serous cystadenocarcinoma of the ovary was studied to determine if patient tumor cells would absorb antiepithelial basement membrane antibody present in patient serum. Absorption of patient serum with trypsinized tumor cells failed to decrease the titer of serum antibasement membrane antibodies. These studies present additional evidence that malignant neoplasms and bullous pemphigoid are not causatively related.


Clinical applications of serum carcinoembryonic antigen and alpha-fetoprotein levels in children with solid tumours. A study was carried out on serum carcinoembryonic antigen (CEA) and alpha-feto-protein (AFP) levels, both measured by radioimmunoassay, in 88 children with malignant solid tumours and in 26 children with nonmalignant disorders, who presented during the years 1973-77. Slightly or moderately raised CEA levels were found at presentation in 11 of 66 children with malignant tumours, in 2 others with recurrent tumours, and in 4 children with nonmalignant disorders. Raised CEA levels generally indicated advanced malignant disease, often affecting the liver, or other hepatic disorders, but were not associated with a specific tumour type. Except in the first months of life, significantly raised AFP levels were detected only in 11 patients with yolk sac-derived tumours, or hepatomas, and in one child with tyrosinosis who later developed a malignant hepatoma. Serial measurements of AFP accurately reflected the clinical response to treatment and in 2 patients indicated recurrence before this could be detected clinically.


Analytical methodology for the determination of Kepone residues in fish, shellfish, and Hi-Vol air filters. The recent discovery of the pollution of the environment with Kepone has resulted in a tremendous interest in the development of residue methodology for the compound. Current multiresidue methods for the determination of the common organochlorinated pesticides do not yield good quantitative analytical results for Kepone. The extracting solvents are usually of insufficient polarity to extract Kepone from the various media. This article describes some of the recently developed methodology for Kepone in air filters, finfish, finfish livers and entrails, shellfish, and archival oyster samples. The finfish and archival oyster samples were Soxhlet extracted using diethyl ether/petroleum ether (1:1 v/v) as the extracting solvent. The finfish livers and entrails were macerated in a Duall tissue grinder containing acetonitrile followed by partitioning of the Kepone into benzene. Shellfish samples were analyzed after extracting the sample with acetonitrile and partitioning the Kepone into benzene. The Hi-Vol air samples were extracted with methanol/benzene (1:1 v/v). The coextracting contaminants were removed by micro-Florisil column chromatography and/or acid digestion. The procedural recovery of Kepone from fortified samples averaged 82%.


["Torsades de pointe" and reentry induced by anti-arrhythmia agents]. Some anti-arrhythmic agents which have a stabilising effect on the membrane, particularly those belonging to Group I of Vaughan William's classification, are likely to cause rythmic disturbances by a re-entry phenomenon (atrial flutter 1/1, premature ventricular contractions, ventricular tachycardia, "torsades de pointe", even fatal ventricular fibrillation). The quinidines have been the most often condemned due to their effect on the action potential and on the speed of conduction. The fact that these re-entry phenomena are more frequent at therapeutic doses than at high or toxic doses suggests an individual susceptibility and the interaction of various predisposing factors: renal insufficiency, potassium depletion, pre-existing conduction disturbances, potentialsing drugs. In order to prevent these incidents it is advisable not to prescribe such anti-arrhythmic agents for susceptible patients, to control individual tolerance and to watch for the first ECG signs of conduction disturbances or of re-entry, such as premature contraction with fixed coupling. Treatment consists in an electrosystolic stimulus and the possible administration of potassium and bretylium.


[The heart diseases of children born to alcoholic mothers]. The authors present their experience of heart disease in children with congenital abnormalities of alcoholic origin, of which they have seen 50 cases in 3 years. This syndrome, which was first described by Lemoine in 1968, appears to be fairly common. It shows the teratogenic properties of alcohol. It is characterised by facial abnormalities which are easy to recognise, gross failure to gain in height and weight, psychomotor retardation of varying severity and several other malformations. From amongst these, cardiac malformations are the most commonly encountered. These are usually in the form of septal defects (atrial and ventricular septal defects).


[Histological studies on the hypothalamic adenohypophyseal-gonadal system after mestranol and chlormadinone acetate administration in fattening bulls]. Seventy-six young steers were examined to elucidate the action of the steroid oestrogen Mestranol and the gestagen chloromadinone acetate upon the morphology of hypothalamus, adenohypophysis, testes, and accessory sexual glands. The doses used were those applicable also to real animal husbandry. The mechanisms of action of the above pharmaceutical products are discussed against the background of the experimental results. No pathological alteration was observed that might prohibit application of the tested sexual steroids to young steers.


[Quantifiable morphokinesis on parts of the hypothalamo-adenohypophyseal-gonadal axis of hormonally castrated boars. I. Studies on boars treated with chlormadinone acetate]. A clearly morphokinetic effect was recorded from the parts of the hypothalamo-adenohypophyseo-gonadal axis in boars that had been treated with chloromadinone acetate (CAP). Luteinising hormone release is inhibited by CAP, which is reflected in an increase of luteinising hormone cells in their storage form. This leads to statistically significant decline in androgenic testicular tissue percentage and in the cell nucleus volumes of Leydig's cells. Excessive weight decrease of the testicle is caused primarily by some 60 per cent reduction in tube length. The neurons of the tested hypothalamic nuclear region, that is the nucleus praeopticus medialis, have shown that in treated animals cell nucleus volumes were significantly depressed.


[Severe viral hepatitis in childhood: course and prognosis]. In 22 children with severe acute viral hepatitis, the course of the disease followed 3 patterns: 8 children completely yielded (regenerative hepatitis); 8 died during the first three weeks of evolution (aregenerative hepatitis); 8 had a prolonged evolution (hyporegenerative hepatitis). In the latter group, 6 patients died after an average survival time of 55 days and 2 patients rapidly developped a cirrhosis. This type of evolution was characterized by persistence of liver failure manifestations, in spite of liver regeneration, as indicated by increased levels of alpha-foetoprotein and presence of pseudo-acini, giantcells and nodules at histological examination. During the second week of evolution, the size of liver, levels of clotting factors VII +X and alpha-foetoprotein concentrations seem to constitute important prognostic factors.


Rapid methenamine silver stain for Pneumocystis and fungi. A rapid modification of the methenamine silver stain for Pneumocystis carinii and fungi employs heat alone to accelerate the silver reduction step and is suitable for both sections and smears.


[Effects of experimental factors on the tinctorial properties of structural elements of the mammalian cerebral cortex (intravital microscopic study)]. Effects of a vital dye--methylene blue--on brain neuronal structures were investigated in the alive preparations of mammal brains by means of contact optical microscope in falling polarization light. Tinctorial properties of neuronal cortical structures were studied under the influence of a number of experimental factors (illumination conditions, dye concentration, temperature regime and oxygen supply to the preparation). A certain dependence in the character of dyeing and bleaching neuronal cells on the factors mentioned above was stated. Conditions for appearance of the alternating photodynamical effect and measures to prevent it were clarified. Practical recommendations were given for application methylene blue in prevital morphological investigations of mammal central nervous system.


[Degeneration of the laminated corpuscles (of Vater--Pacini) following colchicine application to a nerve]. Laminated pacinian corpuscles from the cat mesentery have been studied morphologically and morphometrically after nerve section and colchicine application to the nerve and the results obtained are represented. Similar interventions in the nerve produce changes in the receptors resembling those of wallerian type degeneration, degeneration rate after sectioning being higher than after colchicine application. At early stages after colchicine application the internal cone and its nuclei increase in size. The data obtained suggest the nuclei of the internal cone to be under neurotrophic control of the sensory neuron that might be realized via axoplasmic transport of substances.


[Morphology and morphophysiology of reactive neuronal states]. It is stated that nonspecific reactivity of the neuron to the irritating action of an agent is based on some morphological regularity such as dynamics of its structural-tinctorial changes. There exists a definite parallelism between phase development of the dynamics and fluctuations of the membrane potential (MP): invariable correlation and accompany of these phenomena. Therefore, it is possible to treat MP changes as one of stimulating signs of the neuron and consider them as a component in the complex of its nonspecific changes. The data available in literature and those of the author, make it possible to conclude that the phase of nonspecific changes of the neuron directly affects its specific activity.


The radiotherapy of melanoma.

The radiotherapy of melanoma. A retrospective review of a series of cases of malignant melanoma is reported. Two hundred and forty-six records of patients irradiated from 1958 to 1976 inclusive reveal a sex ratio of males to females of 1.59 to 1, with mean ages of 52.4 years and 54.4 years respectively. Three patients are lost to follow-up. Two hundred and twenty are deceased. Of 23 living, nine are diseased and 14 are well. Of 60 patients treated aggressively, 10 survived five years: one of 26 irradiated only; three of 11 irradiated before surgery; and six of 23 irradiated after surgery. Of 168 treated by radiotherapy for palliation of growth restraint, one died of disease after five years, and two are alive and well after 10 years! The primary site was treated in 20 cases, with three patients surviving five years. Secondary sites were treated in 226 cases, with 10 patients surviving five years.


Interaction of urokinase with alpha2-macroglobulin investigated by isoelectric focusing. Evidence for non-specific dissociable binding. The binding of urokinase to human alpha2M (alpha2-macroglobulin) was investigated in comparison with the formation of the equimolar trypsin-alpha2M complex. Experiments were performed by molecular-sieving on Sephadex G-200, subunit conversion by sodium dodecyl sulphate-polyacrylamide-gel electrophoresis after reduction and isoelectric focusing in linear sucrose gradients with ampholytes pH 3.5-10.0. Urokinase activity was determined with alpha-N-acetyl-L-lysine methyl ester and by activation of plasminogen on unheated fibrin plates. alpha2M was determined by single radial immunodiffusion. alpha2M was capable of binding some urokinase by a non-specific type of attachment that could be disrupted by isoelectric focusing but not by gel filtration. The pI of the undissociated trypsin-alpha2M complex was 6.0, and differed from that of the pure alpha2M (5.2-5.4). Likewise the pI of the immunoreactive alpha2M was 5.2 after exposure to urokinase, whereas the dissociated urokinase focused at pI 10.2. This indicated lack of true inhibitor-complex formation, which was also sustained by total absence of subunit conversion. The results are in agreement with our previous findings with pancreatic and urinary kallikreins.


Contrasting patterns of newer histocompatibility determinants in patients with rheumatoid arthritis and systemic lupus erythematosus. The Ia alloantigens as measures of different alleles of loci in the major histocompatibility complex were determined in patients with systemic lupus erythematosus (SLE) or rheumatoid arthritis (RA). The Ia specificities of reagents used were defined by their pattern of reaction with lymphoblastoid lines derived from normal donors homozygous for HLA-D determinants. The reagent specificities included those associated with a single Dw type as well as those reacting with a single specificity shared by several Dw types. Patients with RA had a marked elevation in the frequency of alloantigens detected by reagent sera that recognize various determinants shared by cell lines from HLA-Dw4, Dw7, or Dw10 individuals (Ia 4-7-10). The frequency of mixed lymphocyte culture alleles Dw4 and Dw10 was found to be increased; however this elevation did not approach the higher frequency for the serologically determined antigens of the Ia 4-7-10 group. In contrast, patients with SLE had an increased frequency of reactions with the reagent alloantisera defined by reactions with either HLA-Dw2 or Dw3 positive cell lines. The data suggest that immunogenetic factors are relevant to both groups of patients, but that these are entirely distinct for each disease.


Immunologic approaches toward detection of type C viral expression in man. Type C RNA viruses have been isolated from a large number of mammalian species. These agents may be horizontally transmitted as infectious cancer-inducing agents, or vertically transmitted from one generation to the next, often in an unexpressed form, within the host genome. To date, the translational products of three viral genes have been identified. With purified virus-coded proteins as probes, sensitive and highly specific radioimmunologic assays have been developed for the detection of antibodies and antigens related to the known type C viruses. These techniques have proved valuable in sero-epidemiologic studies of the horizontally transmitted oncogenic viruses of cats, cattle, and gibbons, and have been used to detect translational products of endogenous viruses in tissues of species from which complete virus has yet to be isolated. This review describes the application of radioimmunoassays in the search for immunologic evidence of type C virus expression in man.


Persistent measles virus infection in vitro and in man. The characteristics of cell lines persistently infected with measles virus are described. Molecular mechanisms for measles virus-persistence in vitro as well as in human disease include defective interfering particles, mutant virus strains, proviral DNA, and nonpermissive cell types. The role of measles virus in subacute sclerosing panencephalitis and evidence for measles virus infection in multiple sclerosis, Paget's disease, and systemic lupus erythematosus are discussed.


[The prognosis of intracranial saccular aneurysms in the conservative treatment (author's transl)]. During the period from June, 1961 to September, 1975, 1080 cases of intracranial saccular aneurysms were experienced in Tohoku University. This investigation is based on the follow-up studies of 59 cases which were treated conservatively. On discharge the condition of the cases were classified into the following 5 groups: "Excellent", the patients is fully capable for employment, "Good", the patient is capable of working, although some neurological deficits remain, "Fair", the patient has one or more of three handicaps--Incapability of walking independently, psychic disturbances or aphasia, "Poor", the patient is completely incapable of walking even with assistance. The results of 59 cases were evaluated on discharge. The total consisted of 15 excellent cases, 2 good, 2 fair, 2 poor and 38 deaths. The follow up studies were obtained from the all cases which were discharged from the hospital (the follow up rate of 100%). The follow-up periods ranged from six months in minimum to 11 years and 9 months in maximum with the average periods of 3 years and 6 months. The results on follow up studies revealed 13 excellent cases, 0 good, 0 fair, 0 poor, and 8 deaths. Six cases out of 13 which were reported as excellent were unruputured cases. The causes of deaths were attributed to rebleeding in 3, progressive deterioration associated with rupture of aneurysm in 2, neoplasma in 1, acute abdomen in 1, and uncertain cause in 1.


Histamine challenge and anterior nasal rhinometry: their use in the assessment of pseudoephedrine and triprolidine as nasal decongestants in subjects with hayfever. 1 Nasal airway resistance (NAR) was measured by anterior rhinometry in ten volunteers with allergic rhinitis. Measurements before and after challenge with three concentrations of histamine diphosphate showed significant rises in NAR for each challenge. 2 In a double-blind, crossover study with the same patients triprolidine (2.5 mg) and pseudoephedrine (60 mg) were shown to be equally effective in reducing the rise in NAR produced by histamine challenge to one nostril; both were significantly better than placebo. 3 The rise in NAR of both nostrils after histamine challenge to one nostril was significantly reduced after pseudoephedrine compared with placebo. This suggests that pseudoephedrine is effective in preventing reflex mucosal congestion in the unchallenged nostril. 4 No increase in the pulse rate or blood pressure of the volunteers was detected after either drug.


Radioimmunoassay of platelet factor 4 and beta-thromboglobulin: development and application to studies of platelet release in relation to fibrinopeptide A generation. Platelet and fibrinogen survival and turnover studies have shown that platelet activation and fibrin formation may occur to different degrees in different thrombotic disorders. More direct evidence of differential involvement of platelet activation and fibrin formation should be provided by specifically measuring the products of these reactions, i.e. released platelet proteins and fibrinopeptide A. Two platelet proteins, platelet factor 4 (PF4) and beta-thromboglobulin (betaTG), were isolated and characterized, and sensitive and specific radioimmunoassays were developed to measure them. These assays were employed, along with the radioimmunoassay for fibrinopeptide A (FPA), to study the release of PF4 and betaTG in relation to FPA cleavage. PF4 and betaTG were released by ADP and collagen with time course and concentration dependence similar to that of [14C]serotonin release. FPA was not cleaved from fibrinogen during ADP or collagen-induced platelet release. Thrombin caused release of PF4 and betaTG as well as cleavage of FPA. Cleavage of FPA occurred with concentrations of thrombin about 100 times less than did release of PF4 and betaTG, and release of [14C]serotinin required still higher thrombin concentrations. Release of [14C]serotonin and platelet proteins was similar as a function of time. Sodium citrate was found to inhibit platelet release induced by thrombin.


Effect of antabuse (disulfiram) on Rous sarcoma virus and on eukaryotic cells. Antabuse (disulfiram) is widely used in the treatment of chronic alcoholism. We have examined the effect of this drug on malignant transformation by Rous sarcoma virus, on eukaryotic cell synthesis, and on nucleic acid binding. It was found that: (1) Disulfiram inhibits the activity of the RNA dependent DNA polymerase of Rous sarcoma virus and inactivates the ability of the virus to malignantly transform chick embryo cells. The monomer of disulfiram, diethyldithiocarbamate does not affect the virus. (2) Disulfiram induced the synthesis of four proteins in normal chick embryo and human foreskin cells. The monomer diethyldithiocarbamate, induced these proteins also. Cellular DNA synthesis is more sensitive to disulfiram than are RNA and protein synthesis. (3) Disulfiram binds to neither DNA or RNA in the presence or absence of copper. However, diethyldithiocarbamate in the presence of, but not in the absence of, copper binds to HeLa cell DNA and to Rous sarcoma virus 70 S genome RNA. These results indicate that this compound, which causes no symptoms in people who do not consume alcohol, may have significant effects on a cellular level.


Analysis of larger than tetrameric poly(adenosine diphosphoribose) by a radioimmunoassay in nuclei separated in organic solvents. Anitibodies were prepared against poly(adenosine diphosphoribose) of an average chain length of 40 adenosine diphosphoribose units by repeated injection of the polymer mixed with methylated albumin and adjuvants into rabbits. The antibody was present mainly in the 7 S fraction of the immunoglobulins. A membrane binding assay was developed, and its specificity determined for the detection of (adenosine diphosphoribose)ngreater than4 in organs. The method is suitable for the study of the variation of the polymer content of nuclei. The size recognition of the anti-poly(adenosine diphosphoribose) globulin fraction was the same for polymers composed of 4--40 adenosine diphosphoribose units, but smaller oligomers were not detectible. A quantitative extraction technique was developed and applied for radioimmunoassay of nuclear (adenosine diphosphoribose)n greater than 4. Organs were freeze-clamped, freeze dried, broken into subcellular fragments in a colloid mill, and the nuclear fraction was subsequently separated in organic solvents in order to preserve the polymer. Nicotinamide and nicotinic acid, when administered in vivo, augmented the (adenosine diphosphoribose)n greater than 4 content of rat liver and heart. Tissues of infant pigeons contained larger quantites of (adenosine diphosphoribose)ngreater than4 than tissues of adult rats.


Template-specific requirements for DNA synthesis by the Mason-Pfizer monkey virus DNA polymerase: unique aspects. The biochemical properties  of DNA polymerase purified from Mason-Pfizer monkey virus were studied, with respect to synthetic and natural template-primer utilization. Thes studies revealed the following new information about the Mason-Pfizer monkey virus enzyme: (a) Mason-Pfizer monkey virus polymerase was found to prefer template: primer molar nucleotide ratios of 2.5-5: 1 for optimal rates of synthesis with poly(C) .(dG)12-18  as template-primer. (b) Poly(A)-directed synthesis was stimulated by the addition of low concentrations of inorganic phosphate to the reaction mixture. (c) Poly(2' -O-methyl-cytidylate), poly(rCm), was the only template studied for which Mn2+ proved the preferred divalent cation. Combinations of divalent cations stimulated rather than inhibited poly(rCm)-directed poly(dG) synthesis by the Mason-Pfizer monkey virus enzyme. (d) Heteropolymeric regions of rabbit globin mRNA and avian myeloblastosis virus 70 S RNA could be copied by the Mason-Pfizer monkey virus polymerase with oligo(dT), oligo(U) or in the case of avian myeloblastosis virus RNA, endogenous primers. In all such studies, Mg2+ was the preferred divalent cation and a distinct preference for the DNA primer in the reverse transcription of natural RNAs was observed. These new findings necessitated comparative studies with the DNA polymerases from Rauscher murine leukemia virus and murine mammary tumor virus, as representative type C and type B retroviruses. Although the Mason-Pfizer monkey virus enzyme was found to share some properties in  common with both type C and type B mammalian viral enzymes, certain of the above properties rendered it unique among the polymerases examined.


Synthesis of Sindbis virus complementary DNA by avian myeloblastosis virus RNA-directed DNA polymerase. Sindbis virus 42 S RNA was efficiently transcribed into complementary DNA (CDNA) by avian myeloblastosis virus alphabeta DNA polymerase using oligo- (dT) or single-stranded calf thymus DNA as primers. Both of the Sindbis virus cDNA products were able to protect 60% of 125I-labeled Sindbis virus RNA, at near equal weight ratios, from RNAase A and T1 digestion. Using hybridization kinetics, the Crt 1/2 value for hybridization of the calf thymus-primed cDNA product with excess Sindbis RNA was determined to be 1.8 9 10-2 mol . s . 1-1. Thes data demonstrate that the Sindbis virus cDNA products are relatively uniform representations of Sindbis virus RNA sequences.


The temperature dependence of the surface tension of aqueous solutions of plasma proteins. The surface behavior of aqueous solutions of fibrinogen, transferrin, gamma-globulin and albumin at the liquid-gas interface has been investigated by a modified Wilhelmy technique. The temperature dependence of the surface tension was studied over a temperature range of 20--80 degrees C and a pH range of 2--12. Most pronounced conformational changes of fibrinogen with this technique were found in physiological conditions: 35--45 degrees C and pH 7--8. A conformational change was found for gamma-globulin and transferrin solutions, but at a higher temperature and less pronounced than fibrinogen. Albumin did not undergo conformational transitions to a significant extent.


Bovine fetus-specific serum proteins. Purification and characterization of alpha1-fetoprotein and immunological identification of alpha2- and beta-fetoproteins. Bovine alpha1-fetoprotein was isolated from fetal calf serum by successive procedures of concanavalin A-Sepharose chromatography, DEAE-Sephadex chromatography, SP-Sephadex chromatography and preparative disc polyacrylamide gel electrophoresis. The bovine alpha1-fetoprotein preparation was considered homogeneous by several physicochemical and immunochemical criteria. Bovine alpha1-fetoprotein has a molecular weight of 68 000 with an amino acid composiotn similar to that of other mammalian alhpa1-fetoprotein. In addition, bovine alpha1-fetoprotein was shown to exist as two distinct variants on the basis of carbohydrate heterogeneity. alpha2-Fetoprotein and a new beta-fetoprotein were immunologically identified in fetal calf serum. These fetoproteins, like alpha1-fetoprotein, were not detectable in non-pregnant cow serum by immunoelectrophoresis.


A catalytic role for heparin. Evidence for a ternary complex of heparin cofactor thrombin and heparin. The interaction of heparin with chemically modified thrombin and heparin cofactor is studied. Amidinated heparin cofactor does not bind to heparin-agarose and the reaction rate of the amidinated inhibitor with unmodified thrombin is not affected by heparin. Likewise, thrombin modified with 1,2--cyclohexanedione does not bind to heparin agarose and the reaction rate of the modified enzyme with unmodified inhibitor is not affected by heparin. In the absence of heparin, the modified and unmodified proteins react at the same rate in all possible combinations. Affinity chromatography of diisopropylphosphoryl thrombin on heparin cofactor coupled to Sephadex G--50 is used to study the binding of heparin cofactor and thrombin to heparin. The thrombin for all experiments is tritium-labeled and then inactivated with diispropylfluorophosphate. Thrombin is not bound to heparin cofactor-Sephadex columns. However, after treatment of the columns with a heparin solution, thrombin binds tightly, and is eluted at high ionic strength. Bound thrombin can also be eluted with either excess non-radioactive thrombin or excess free heparin. Heparin-dependent binding of thrombin does not occur if the heparin cofactor-Sephadex is heat-denatured. The ability of heparin to couple solution-phase thrombin to solid-phase heparin cofactor indicates that a ternary complex is formed. Analysis of the binding of the proteins to heparin by a dye displacement method suggests that at least one site on heparin binds to thrombin but not to heparin cofactor. Further support for a catalytic role for heparin derives from the ability of catalytic concentrations of heparin to enhance the rate of hydrolysis of prothrombin by thrombin, another protein pair which bind mutually to heparin.


Elimination of asialofetuin and asialoorosomucoid by the intact rat. Quantitative aspects of the hepatic clearance mechanism. The capacity of the liver to eliminate asialofetuin and asialoorosomucoid was investigated in intact rats. From plasma radioactivity curve measurements and assays on tissue homogenates the liver is shown to be able to dispose of an average of 19.8 microgram of asialofetuin/min per 100 g body weight. No other major route is identified for the disappearance of asialofetuin from the plasma, although trace amounts of the protein were detectable in the urine. From analyses of the plasma radioactivity curves the elimination process for asialoorosomucoid appears to be comparatively complex because of the existence of extrahepatic disposal routes. Quantification of labelled asialoorosomucoid in liver homogenates indicates, however, that the hepatic clearance rate for asialoorosomucoid is similar to that for asialofetuin. Urinary excretion significantly contributes to the disappearance of asialoorosomucoid from the plasma but the hepatic and renal routes do not account for all the protein lost from this compartment. At plasma concentrations above the maximal eliminative capacity of the liver, the hepatic clearance of asialofetuin obeys zero-order kinetics and is remarkably constant. Elimination of a quantity of asialoglycoprotein which exceeds the calculated total number of binding sites in the liver does not reduce the efficiency of the pathway, and studies of [3H]leucine incorporation indicate that the lectin, unlike the bound asialoglycoprotein, is not destroyed in the elimination process. Cytochalasin B (80 microgram/100 g body wt.) had no measureable effect on the hepatic clearance of asialofetuin. Administration of colchicine (10 mg/100 g body wt.) resulted in transitory accumulations of asialoorosomucoid in the liver, presumably due to interference with the intracellular transport of the endocytised protein.


[Effect of inductors of alkali cation permeability on cytochrome c bound to mitochondrial membrane]. The effect of inductors of alkali cation permeability--valinomycin, gramicidin A, gramicidin S and its N,N'-diacetyl derivative--on rat liver mitochondria during respiration has been studied. It is shown that valinomycin, gramycidin A and diacetylgramicidin S at optimal concentration for uncoupling cause two-phase activation of mitochondrial respiration and that this effect results from cytochrome c solubilization. Gramicidin S at optimal concentration cannot remove cytochrome c from the respirating mitochondria. It is suggested that this property of gramicidin S is owned to cytochrome c immobilisation in membrane, due to the effect of this compound.


A case of slow variant bisalbuminmia: study of its composition in amino acids. The authors report an observation of slow variant bisalbuminemia. After electrophoretic separation of two varieties, the amino acid chromatographic analysis has permitted them to show that in the slow variant the molecule of albumin has undergone a substitution of several glutamic acid residues by lysine and of several alanine residues by proline. Such an alteration poses the problem of its effect on the antigenic specificity of the molecule and that of its mechanism of production.


Hydrodynamic orientation of sperm heads for flow cytometry. Two specially shaped sample injection tubes orient flat-shaped mature sperm heads in a flow cytometer. Orientation allows control of a photometric difficulty experienced with some flow cytometers in the measurement of fluorescent and scattered light from sperm and other flat cells. Both relative DNA-stain content and a measure of cell flatness can now be determined simultaneously for each cell at the high rates possible with flow instruments.


[Effect of antibodies to denatured DNA on the human bone marrow cells forming colonies in semi-solid agar]. gamma-globulin from rabbit sera containing antibodies to denatured DNA or cytidine suppressed the efficiency of colony formation in agar cultures of human bone marrow cells. The antibodies to DNA isolated from the immune sera by the immunosorbent produced an analogous effect. gamma-globulin from the sera of intact animals and immune sera after the removal of antibodies to denatured DNA from them produced a different effect. gamma-globulin in a concentration of 0.28 and 5 mg failed to influence the colony-formation of 2.10(5) explanted nucleus-containing cells, by stimulated the colony growth when added in a concentration of 15 mg.


[Histamine-liberating effect of antihistaminics on the isolated rat mast cells]. Research Allergological Laboratory of the USSR Academy of Medical Sciences and Laboratory of Pharmacology, S. Ordzhonikidze Research Chemico-Pharmaceutical Institute, Moscow. Among the tested new antihistaminic drugs (quinuclidine derivatives) quinuclidyl-3-(O-tolyl) carbinol possessed histamine releasing action (HRA) on the isolated rat mast cells. In used concentrations (up to 0.4 mmol) all phenothiazines (promethazine, phenethazine, chlorpromazine, methylene blue) had HRA. There was no correlation between the HRA and the antihistaminic activity of the tested drugs. Histamine release induced by antihistaminic drugs and a steep dose-response curve, was produced at low temperature and was not inhibited under conditions of inhibition of energy-dependent stage of 48/80-induced histamine release. It was concluded that the tested antihistaminic drugs which had HRA were non-selective histamine releasers.


[Expression of C-type oncornavirus proteins in tumors of CC57BR mice]. Murine endogenous oncornavirus (C-type) genome expression was investigated in methylcholanthrene-induced (MC-induced) tumours of low-leukemic CC57BR mice by means of the radioimmunodiffusion method, using the precipitating test-systems. The gs-1 antigen of murine C-type viral p30 protein and Gross leukemia virus type-specific antigen (AGLV) served as viral genome markers. The gs-1 antigen was found in the primary and continuous MC-induced sarcomas, whereas AGLV appeared only during the tumour passage. A distinct association was revealed between the quantity of p30 protein (gs-1 antigen titre) and the presence of AGLV. Simultaneously with the appearance of AGLV the gs-1 titre increased markedly and the AGLV disappearance was always accompanied by a decrease of the gs-1 titre. The data presented suggest a coordinated expression of the two investigated proteins of murine C-type endogenous oncornaviruses in the chemically-induced tumours of CC57BR mice.


[Effect of alpha fetoprotein on the processes of reparative regeneration of the skin and tubular bones]. The influence of alpha-fetoprotein of man, dog and cat on the processes of reparative regeneration of the skin and long tubular bones was studied. alpha-fetoprotein was shown to stimulate regenerative processes by forming young connective tissue, but had no strong species specificity. Commercial preparation of human alpha-fetoprotein proved to possess maximum physiological activity in comparison with other alpha-fetoproteins under study.


[Thermostable antigens of malignant tumors and normal tissues of experimental animals]. Gel precipitation and immunoelectrophoresis tests tests demonstrated the presence of thermostable antigens in dimethylbenzanthracene-induced malignant tumours of the muscle tissues of Wistar rats. These antigens were relatively tissue-specific for the tumours, amniotic fluid and the uterus on the one hand, and for the lung, spleen, and the serum on the other hand. The thermostable antigens of the tumours and the amniotic fluid had the electrophoretic mobility in the zone of alpha and alpha1-globulins.


Edinger-Westphal nucleus: projections to the brain stem and spinal cord in the cat. The efferent connections of the Edinger-Westphal (EW) nucleus of the cat have been examined using the autoradiographic anterograde axonal transport technique. Following injections of [3H]amino acids into the EW nucleus, fibers could be traced from this region to a number of sites in the caudal brain stem and spinal cord heretofore not known to receive an afferent input from this nucleus. Two descending pathways have been identified. One pathway travels in the medialmost aspect to the medial longitudinal bundle and terminates in the dorsal accessory olive. The other pathway leaves the nucleus laterally, coursing through the medial tegmentum, and then shifts to a ventrolateral position in the rostral rhombencephalon. Some fibers of this lateral pathway curve dorsally and terminate in the medial parabrachial nucleus. The remainder of this fiber system lies ventral to the spinal trigeminal complex, with some axons terminating in the subtrigeminal nucleus, while other fibers continue ventral to the caudal part of the spinal trigeminal nucleus and appear to terminate in the marginal layer and ventromedial part of this nucleus. Another component of this system terminates between the gracile and medial cuneate nuclei. The main pathways from the EW nucleus to the spinal cord include (1) some fibers which course through the dorsal column nuclei into the ventromedial part of the dorsal columns, and (2) other fibers which continue caudally immediately along the ventrolateral aspect of the spinal trigeminal nucleus and then proceed to the spinal cord in the region between the dorsal horn and the lateral cervical nucleus. These EW fibers appear to terminate mainly in Rexed's lamina I (marginal layer); other fibers, from both tracts appear to terminate in lamina V. There was no evidence for any ascending projections from the EW nucleus. Confirmatory data were obtained from a series of horseradish peroxidase (HRP) experiments, in which EW neurons were retrogradely labeled following injections into the dorsal column nuclei, spinal trigeminal nucleus, inferior olivary nucleus, or spinal cord. These results clearly indicate that the traditional view of the EW nucleus as merely a parasympathetic preganglionic nucleus should be seriously questioned.


Neuronal, non-neuronal and hybrid forms of enolase in brain: structural, immunological and functional comparisons. Three forms of the glycolytic enzyme, enolase [2-phospho-D-glycerate hydrolase (E.C. No. 4.2.1.11)] have been prepared from rat whole brain extract. The most acidic enolase form is neuron specific enolase (NSE) which had previously been designated neuron specific protein (NSP). The least acidic form designated non-neuronal enolase (NNE) has been purified and compared structurally, immunologically and functionally to NSE. NNE is a dimer of 86,500 M.W. consistint of two very similar subunits. The data establish that NNE is larger than NSE which has been shown to be composed of two apparently identical 39,000 molecular weight subunits (78,000). NNE is less acidic than NSE having a pI of 5.9 compared to the value of 4.7 for NSE. Structural and immunological analysis establishes that the NNE subunit is distinct from the NSE subunit, and are therfore products of two separate genes. The structural designation of NSE is (gammagamma) and that of NNE (alpha' alpha'). NSE is strictly localized in neurons indicating that the gene coding for the gamma subunit is only expressed in neuronal cells. The intermediate brain enolase form has been partially purified; structural and immunological evidence indicate that it is a hybrid molecule consisting of one NNE subunit and one NSE subunit (alpha'gamma).


Axonal transport of [3H]proteins in a noradrenergic system of the rat brain. Proteins synthesized from [3H]leucine injected into the rat nucleus locus coeruleus (LC) were transported through the hypothalamus in 5 successive waves at rates of 72-192, 24-48, 13-20, 3-4 and 1.4-2.9 mm/day (waves I through V, respectively). Waves I through IV began axoplasmic transport in the LC within the first few hours after [3H]protein synthesis began in the LC. Wave V was delayed in onset for 1.7-3.7 days and was also probably transported through the contralateral hypothalamus. Wave IV was not transported within the LC-hypothalamic axons ascending through the dorsal noradrenergic bundle since its transport was not blocked by 6-OHDA lesions in this bundle, as was transport of the other 4 waves. Unilateral dorsal bundle lesions caused a well defined caudal backup of dopamine-beta-hydroxylase immunofluorescence and a fall in dopamine-beta-hydroxylase activity in the ipsilateral frontal cortex and hypothalamus of 55% and 9%, respectively. Bilateral lesions caused only a significantly further reduction in hypothalamic levels indicating crossed innervation of the hypothalamus by the LC of 27%. Waves I and II have been classified as rapid transport and contained 33% of the transported [3H]protein. Wave V was slowly transported and contained 51% of the transported [3H]protein, while wave III was intermediate in rate and contained 16% of transported [3H]proteins.