Thermal destruction of Clostridium botulinum spores suspended in tomato juice in aluminum thermal death time tubes. The heat destruction characteristics of Clostridium botulinum spores suspended in tomato juice and phosphate buffer were determined by the survivor curve method with aluminum thermal death time tubes. Two type A strains of C. botulinum and a type B strain were evaluated. Strains A16037 and B15580 were implicated in outbreaks of botulism involving home-canned tomato products. Strain A16037 had a higher heat resistance than either 62A or B15580. The mean thermal resistance (D-values) for A16037 in tomato juice (pH 4.2) were: 115.6 degrees C, 0.4 min; 110.0 degrees C, 1.6 min; and 104.4 degrees C, 6.0 min. The mean D-values for A16037 in Sorensen 0.067 M phosphate buffer (pH 7) were: 115.6 degrees C, 1.3 min; 110.0 degrees C, 4.4 min; and 104.4 degrees C, 17.6 min. At each test temperature, the D-values were approximately three times higher in buffer than in tomato juice. The z-value for C. botulinum A16037 spores in tomato juice was 9.4 degrees C, and in buffer the z-value was 9.9 degrees C. The use of aluminum thermal death time tubes in a miniature retort system makes it possible to determine survivor curves for C. botulinum spores at 121.1 degrees C. This is possible because the lag correction factor for the aluminum tubes is only about 0.2 min, making possible heating times as short as 0.5 min.


Effect of dredge spoil deposition on fecal coliform counts in sediments at a disposal site. The most-probable-number of fecal coliforms in sediments was monitored at the New London dump site in Long Island Sound during the deposition of dredge spoil from the Thames River. Although the geometric mean for fecal coliforms at five stations in the river was 14,000/100 ml before dredging commenced, the deposition of this material did not increase the incidence of fecal coliforms at 17 spoil stations and 13 control stations in the disposal and surrounding areas. Fecal coliforms appear to occur only in the surface sediment material and are diluted by the subsurface material during the dredging operation. Fecal coliform analyses of bottom waters during high and low tides indicated that the flow of water from the Thames River played a major role in determining the most-probable-number of fecal coliforms in the sediments at the disposal site.


Membrane filter technique for the quantification of stressed fecal coliforms in the aquatic environment. A two-layer membrane filtration (MF) medium (injury-mitigating MF [IM-MF]) and a procedure for the enumeration of injured fecal coliforms are described. These procedures included the addition of glycerol and acetate plus reducing agents to both layers of a two-layer medium and rinsing of the filter with a rich resuscitation medium. Some changes in incubation time and temperatures were used. This method was compared with the multiple-tube fermentation most-probable-number procedure and the one-step M-FC agar-membrane filter method (direct M-FC) in terms of fecal coliform recovery from various aquatic environments that cause bacterial injury. With chlorinated sewage effluents, results of the IM-MF technique were equal to or greater than the most probable number in 9 of 18 trials and were 1.3 to 19 times greater than the M-FC method. When sewage samples were chlorinated in the laboratory, fecal coliform counts with IM-MF equaled or exceeded the most probable number in 7 of 15 trials and always exceeded the M-FC. M-FC was exceeded by IM-MF in 30 of 33 trials with clean mountain stream water. Fecal coliform bacteria that were exposed to low levels of an iodophore in the laboratory produced IM-MF counts 3 to 10 times greater than those with M-FC. A biochemical rationale for the formation of the IM-MF medium is discussed.


Comparison of the surface structure, metal binding, and fecal coliform recoveries of nine membrane filters. A comparative study was made of nine commonly used membrane filters from five manufacturers, all recommended for enumeration of coliform bacteria. Bacterial recoveries and flow rates were examined from three types of water and were found to correlate with the surface pore structure determined by scanning electron microscopy. The sorption of metals was also determined. The results of these studies indicate that the five best membranes for fecal coliform recovery could be placed in two groups: Millipore HC and Gelman, followed by Johns-Manville SG and AG and Sartorius 13806.


Aflatoxin localization by the enzyme-linked immunocytochemical technique. Visualization of aflatoxin deposits within Aspergillus parasiticus ATCC 15517 hyphae was made possible by using an enzyme-linked immunocytochemical technique.


Enzyme-linked immunosorbent analysis for aflatoxin B1. An enzyme-linked immunosorbent analysis (ELISA) permitted the detection of less than 10 pg of aflatoxin B1 per ml. The antitoxin was most specific for aflatoxins B1 and B2alpha, and least specific for aflatoxin G1.


Proteolytic mutants obtained from Clostridium botulinum type E. Proteolytic mutants were isolated from toxigenic strains of Clostridium botulinum type E after several transfers. When these cultures were plated on blood agar, almost all of the colonies obtained were proteolytic, and there were fewer toxigenic colonies than nontoxigenic colonies. The proteolytic mutants and nonproteolytic original strains were different in their biological properties.


Bullous pemphigoid. Clinical and immunologic follow-up after successful therapy. Thirty-six patients with bullous pemphigoid (BP) have been periodically evaluated for four years. This study demonstrates that BP may occur in a transient predominantly localized form that remits spontaneously, and most BP patients after successful therapy remain in prolonged clinical remission. In this study, all patients with active or recurrent disease had IgG and/or C3 basement membrane zone (BMZ) deposition. Serum anti-BMZ antibodies was an inconstant feature. In most instances, clinical remission of BP was associated with disappearance of BMZ ig and C3 deposition and serum BMZ antibodies. Fluorescein-conjugated, antihuman C3 appears to be a more sensitive immunoreagent than antihuman, class specific, immunoglobulin antisera in detecting positive BMZ staining in BP. Combined therapy with azathioprine plus prednisone appears to be superior to prednisone alone in the treatment of BP.


Defective monocyte chemotaxis in mycosis fungoides. Monocyte-macrophage function was studied in 14 patients with histologically confirmed mycosis fungoides. Three component parts of the monocyte-macrophage system were examined. Monocyte bacterial phagocytosis and bacterial killing were normal. However, a defect in directional mobility (chemotaxis) was found in patients with mycosis fungiodes (P less than .0025) when compared to 35 healthy controls. Defective chemotaxis was present regardless of disease stage or therapy. This defect in monocyte chemotaxis represents a previously unrecognized immune deficiency and may help explain the frequent infections found in patients with mycosis fungoides.


Immunopathologic study of herpes gestationis in mother and infant. In vivo bound C3 was demonstrated by immunofluorescence in lesions of both maternal and infant skin in an immunopathologic study of herpes gestationis. Immunoglobulins and other complement components of both classical and alternative pathways were not found. The serum was also negative for circulating antibody to basement membrane of skin. Maternal serum had a factor present that was able to deposit C3 from fresh normal serum onto the basement membrane zone of skin. Hemolytic complement titrations showed a decrease of CH50 levels in serum of both classical and alternative pathways in the mother and the classical pathway in the infant, with a decrease of C2 and C4 in both. To our knowledge, this is the first reportof a decrease in serum complements in herpes gestationis and indicates that complement may not be activated locally.


Eruptive nevocytic nevi after severe bullous disease. In two patients, hundreds of nevocytic nevi (melanocytic nevi) appeared in sites of severe bullous dermatoses that were considered to be toxic epidermal necrolysis and erythema multiforme exudativum, respectively. This phenomenon may result from benign melanocytic hyperplasia accompanying keratinocytic hyperplasia during the healing process of some of the denuded areas of the skin.


The immunoglobulin-bearing cells in the lamina propria and the clinical response to a gluten-free diet in dermatitis herpetiformis. In 17 patients with DH, multiple duodenal and jejunal biopsies were performed. In all patients the small-intestinal biopsy-specimens showed histopathological changes compatible with coeliac disease. Fourteen of the patients maintained a gluten-free diet (GFD) for more than 8 months. The small-intestinal lesions improved in all patients investigated during the GFD. The dosage of Dapsone needed to control the sin lesions could be reduced by more than 50% in 4 patients and the Dapsone could be stopped in 5 other patients on GFD. The immunoglobulin-bearing cells in the lamina propria were counted in 8 patients not on a gluten-free diet, in 6 patients on gluten-free diet, and in 8 healthy controls. The numbers of IgA-, IgM- and IgG-bearing cells were increased in most of the DH patients who were not on a gluten-free diet. The number of IgM-bearing cells in the DH patients who were on a gluten-free diet was the same as that in the control group. This may indicate a mainly IgM response in the lamina propria induced by gluten.


[Fibrin depostition in granuloma anulare (author's transl)]. In 6 cases of granuloma anulare direct immunofluorescence studies were carried out with anti-fibrinogen and anti-immunoglobulin sera. Fibrin deposition was found in all lesions. The result is discussed with reference to local fibrinolysis.


The effect of serotonin on pepsin inhibition by duodenal fat. The purpose of these studies was to evaluate the possible role of serotonin on pepsin secretion. The results of our data suggest that serotonin is a stimulator of pepsin secretion since (a) pretreatment with reserpine (which depletes serotonin stores) abolished the stimulatory effect of histamine on pepsin secretion; (b) pretreatment with UML-491 (a serotonin blocking agent at the effector level) augmented the pepsin inhibition induced by the intraduodenal infusion of fat, and (c) 5-hydroxytryptophan (a serotonin precursor) caused a significant stimulation of pepsin secretion. The mode of action of serotonin is probably by a direct hormonal action on the gastric chief cells.


Observations on preservation, bile drainage and rejection in 64 human orthotopic liver allografts. The combined Cambridge/King's College Hospital series of 64 orthotopic liver grafts' experience dates back to 1968. Many patients were referred for liver grafting late in the course of their diseases and were operated on when they were too ill or died before suitable livers could be found. Complications of biliary drainage were the most frequent causes of death. In the past two years we have acquired experience of a method of liver preservation which permits up to 8 hours of safe storage without any complicated machines and we have been able to transport 22 livers by air and road from other institutions to our own unit. This has enlarged the pool of potential donors and therefore reduced the average waiting time for a liver transplant. A new method of biliary drainage has been employed in 24 patients using the vascularised gall bladder as a conduit between the donor and recipient common ducts. This has resulted in a marked reduction in early postoperative complications of biliary fistula and obstructed bile drainage. Now that survival has improved, it has become clear that uncontrollable rejection of the allografted liver in man is much less severe than that experienced with kidney transplants. The results of liver grafting in our unit during the past year are better than those obtained with kidneys from cadaver donors.


Epidemiology of venous thromboembolism.

Epidemiology of venous thromboembolism. This review of the epidemiology of venous thromboembolism includes estimates of incidence and prevalence of venous thrombosis and its sequelae, a discussion geographical, annual and seasonal variations and data concerning possible risk factors. Selection of patients at increased risk for development of deep venous thrombosis or pulmonary embolism for specific diagnostic screening or for prophylactic therapy with low-dose heparin may be a more effective approach to lowering morbidity and mortality from this disease.


Phenothiazine-induced ECG abnormalities: effect of a glucose load. A total of 54 schizophrenic patients, 27 male and 27 female, satisfying study criteria, were randomly assigned to one of three treatments: placebo; perphenazine, 20 mg/day; or the combination of amitriptyline, 125 mg/day, with perphenazine, 20 mg/day. Medication was administered under double-blind conditions for 12 weeks, after which ECGs were taken following an overnight fast and again following a 600-calorie meal. Among patients receiving perphenazine or amitriptyline-perphenazine, there was a statistically significant increase in repolarization abnormally after eating, whereas placebo-treated patients incurred no such increases. This supports the hypothesis that phenothiazine-induced ECG changes may be caused or facilitated by the glucose load. The incidence of increase in repolarization abnormality after the meal was higher among female patients than among male patients. The findings are of practical significance for readings of abnormality in the ECG of phenothiazine-treated patients.


Comparative studies on the immunologic properties of free endotoxin and lipopolysaccharide from Shigella sonnei. Immunogenic, antigenic and toxic properties of free endotoxin isolated from culture media of Shigella sonnei, phase I, was studied. Experiments with mice showed that free endotoxin is about 3--4 times more active immunogenically and twice as toxic as cellular lipopolysaccharide. On the other hand, the antigenic activity of free endotoxin was about equal to that of lipopolysaccharide. In addition, free endotoxin had a common property characteristic for endotoxins isolated from Gram-negative bacteria, namely it stimulated the immunologic response to unrelated antigens, e.g. antigens of sheep red blood cells.


Studies on the activity of antithymocyte serum (ATS), particularly on the relation between its anti-inflammatory and immunosuppressive effects. Anti-inflammatory and immunosuppressive activity of equine antirabbit antithymocyte serum (ATS) was studied. ATS lowered serum levels of complement, inhibited cell proliferation in the granulation test, and prolonged survival of allogeneic skin grafts. Its action was accompanied by lymphopenia. Repeated immunization enhanced the activity of ATS in the graft rejection test and the drop in serum complement, and at the same time attenuated the effect on the granulation process and lowered proteolytic activity.


Intralysosomal lipid in long-term maintenance transplant atherosclerosis. Intralysosomal accumulation of lipid has been implicated as an important mechanism in the pathogenesis of atherosclerosis. Although atherosclerosis develops frequently in organ transplants maintained on a long-term basis, to our knowledge no studies to date have demonstrated the intracellular localization of the lipid in this setting. Light and electron microscopic study of a renal artery branch from a transplanted kidney maintained for 3 1/2 years demonstrates that the lipid is sequestered within intimal smooth muscle cell lysosomes. The features of the atherosclerotic plaque in long-term transplantation appear to be identical to spontaneous lesions or those induced experimentally.


Bilateral intraocular Nocardia asteroides infection. A 38-year-old man with hypogammaglobulinemia and pulmonary Nocardia asteroides infection developed an intraocular Nocardia infection. The diagnosis was confirmed by examination of a specimen removed at pars plana vitrectomy. The chorioretinal infection in one eye resolved partially, with no organisms visible on histopathologic examination of the globe at autopsy. One month before the patient's death from disseminated nocardial infection, the previously uninvolved right eye developed a new metastatic nocardial chorioretinal lesion. This lesion rapidly progressed in size until the patient's death and showed on postmortem examination the presence of organisms characteristic of N asteroides.


Tarsal pedicle flap for lower eyelid reconstruction. When reconstructing the lower eyelid with use of a temporal-zygomatic skin flap, the inner lining can be provided by a tarsal pedicle flap from the upper eyelid. The flap is hinged in the superior cul de sac and is similar to the letter T on its side, obviating the need to use nasal septal cartilage and mucosa. This method maintains the upward curve of the lower eyelid and restores the lateral canthal angle while causing very little noticeable shortening of the horizontal fissure.


[Biocompatibility of wood in bone tissue (author's transl)]. Alcohol-extracted and sterilised cylinders of ash- and birchwood were implanted into the tibia of rabbits. The selection criteria of these two wood species were thoroughly described. 28 implants were evaluated after 3, 5, 14 and 32 weeks using undecalcified microtome sections and ground sections employing polychromatic sequential-labelling and microradiography. In spite of a foreign-body-tissue reaction new bone is formed around the implants and within the pores of the wood as well. Both wood species therefore seem to be suitable implant materials, which were anchoraged by the bone itself. A tensile strength and elastic properties comparable to bone and simple handling would be the advantages of this material.


Porosity in gold cast against an electroformed gold matrix in an inlay technique. The information obtained from the metallographic procedure used in this paper, as compared with techniques of the past, indicates that certain subgroups such as pinhole porosity, microporosity and subsurface porosity require re-evaluation.


Anterior tooth positions in prosthodontics. The problems associated with positioning anterior health teeth in prosthetic treatment procedures are discussed and some recent work reviewed. Useful guides for establishing the artificial anterior dental relationship have been described and an application of these guides has been illustrated.


The clinical development of the glass-ionomer cement. II. Some clinical applications. A description is given of the varied clinical applications of this cement; these applications include its use as a filling material, as a sealant and as a luting agent.


Secondary intention post and core.

Secondary intention post and core. Methods for restoring existing crown and bridge restorations to normal function following pulp death are reviewed. Endodontic therapy must be supplemental by a reinforcing post-core system if the crown or bridge is to withstand normal functional loads indefinitely.


The stability of maxillary expansion.

The stability of maxillary expansion. The assessment and treatment by rapid maxillary expansion of 516 cases referred by medical practitioners for the treatment of poor nasal airway is reported. The technique is simple and most suitable for the correction of crossbite and Class III malocclusion and for establishing a nasal airway in habitual mouth breathers with a history of ENT problems.


Tissue antigens: autoantigens, alloantigens, xenoantigens and neoantigens. The subject of Clinical Immunology is developing hand in hand with a wide and rapidly moving area of laboratory technology. The result is a better understanding of autoimmune disease, tissue transplantation rejection, foetal-maternal incompatibility, allergic disease, immunodeficiency disorders, adverse reactions to drugs, aberrant responses to bacterial and viral infections, and growth and spread of malignant cells. Basic to this understanding is the need to appreciate the character and composition of natural substances which act as immunogens and elicit antibodies. These substances have, according to their origin, been classified as autoantigens, alloantigens, xenoantigens and neoantigens. This review summarizes our knowledge relating to such antigens, emphasizing those aspects relevant to human disease and pointing to the major gaps that future research must bridge.


Inefficiency of sanitation measures aboard commercial aircraft: environmental pollution and disease. Recent investigations at Tokyo International Airport have proven that environmental pollution resulting from the inefficient disposal of human excretion aboard aircraft is an important problem from the standpoint of quarantine. It is, therefore, recommended that the worldwide aviation industry take immediate measures to improve conditions and eliminate this problem, which has thus far been ignored by aircraft designers, airport administration, and CAB personnel.


Immunomorphologic methods in routine pathology. Application of immunofluorescence and the unlabeled antibody-enzyme (peroxidase-antiperoxidase) technique to formalin fixed paraffin embedded kidney biopsies. Immunofluorescence and the unlabeled antibody enzyme (peroxidase - antiperoxidase) method were applied to formalin fixed conventionally paraffin-embedded kidney biopsy material, and optimal working conditions are reported. Both methods were suitable for the demonstration of immunoglobulin and complement deposits within glomerula. They were equally sensitive on dewaxed pronase-treated sections. Pronase treatment decreased nonspecific background fluorescence and increased the sensitivity of both immunomorphologic techniques, possibly by enhancing antigenicity.


Chitin biosynthesis in protoplasts and subcellular fractions of Aspergillus fumigatus. The biosynthesis of chitin has been obtained in broken mycelia and protoplasts of the fungus Aspergillus fumigatus. The specific activity of chitin synthase (EC 2.4.1.16) in a membrane preparation from protoplasts derived from the hyphal tips of A. fumigatus was 26.8-fold greater than that of the chitin synthase in broken mycelia, indicating that the active chitin synthase is located primarily in a membrane-bound site at the hyphal tip. Polyoxin D was a potent competitive inhibitor of the enzyme, having Ki 5.2 +/- 0.8 micron with respect to the natural substrate UDP-N-acetyl-D-glucosamine, which has Km 1.58 mM.


Purification and steady-state kinetics of adenosine 5'-pyrophosphate sulphurylase from baker's yeast. ADP sulphurylase (EC 2.7.7.5) was purified by chromatography on Sephadex G-200 and DEAE-cellulose. The enzyme was assayed by measuring the incorporation of [32P]Pi into ADP in the presence of the substrate for the reverse reaction, adenosine 5'-sulphatophosphate. In the concentration ranges investigated, by using initial-velocity, product-inhibition and isotope-exchange studies, the data were consistent with a Ping Pong reaction mechanism, with Km for adenosine 5'-sulphatophosphate of 1.20 +/- 0.08 mM and a Km for Pi of 4.95 +/- 0.15 mM. Competitive substrate inhibition by Pi (Ki = 11.7 +/- 0.3 mM) was found. ADP sulphurylase catalyses a sulphate-independent Pi-ADP exchange reaction, the kinetics of which are consistent with the kinetics of the overall reaction, inconsistent with the assay of Burnell & Anderson [(1973) Biochem. J. 133, 417-428], which is based on a sulphate-dependent Pi-ADP exchange reaction.


Evaluation of a new method for the continuous measurement of the distribution of the blood flow between the two lungs. The magnitude of the pulmonary vasoconstrictor response to alveolar hypoxia may be studied by measuring the reduction in blood flow when one lung is ventilated with nitrogen. A radioisotope technique has been developed which permits the distribution of blood flow between the two lungs to be measured continuously. This technique was evaluated in dogs and found to be satisfactory.


Glycopyrrolate-neostigmine mixture for antagonism of neuromuscular block: comparison with atropine-neostigmine mixture. Glycopyrrolate, a new anticholinergic agent, was evaluated and compared with atropine. Glycopyrrolate 0.2 mg to neostigmine 1.0 mg was found to be safe and effective. The heart rates remained more stable with glycopyrrolate, and the frequency of arrhythmia, which was both transient and of no consequence, was similar in the two groups. The antisialogogue action of glycopyrrolate was superior to that of atropine.


Facilitation of nodal metastasis from a non-immunogenic murine carcinoma by previous whole-body irradiation of tumour recipients. Of 193 CBA mice kept under prolonged observation after excision of small intradermal transplants of a non-immunogenic tumour (CBA Carcinoma NT), 27 (14%) presented with local recurrence, 19 (10%) with regional lymphnodal metastasis (RNM) and 72 (37%), with pulmonary metastasis +/- other systemic metastases. When mice were exposed to sublethal whole-body irradiation (WBI) before tumour transplantation, the incidence of RNM rose to approximately 80% and the latent period was reduced from approximately 60 days to approximately 40 days after tumour transplantation. This enhancement of RNM by WBI was undiminished when the interval between WBI and tumour transplantation was increased from 1 to 90 days. An explanation for this effect in terms of immunosuppression by the WBI is unlikely for the following reasons: the tumour was non-immunogenic by standard quantitative tests; the effect persisted long after the expected time for recovery of immune reactivity; and i.v. injection of normal marrow and lymphoid cells after WBI failed to reduce the effect. That the effect was systemic was proved by failure of local pre-irradiation of the tumour bed or regional node to enhance RNM. The effect was not observed when WBI was given 4 days after excision of tumours. These and other experiments failed to indicate the mechanism of the effect of WBI, but its long persistence suggests that it may relate to stored lethal radiation damage in migrating cells of slow turnover tissues.


Lymphocyte markers in non-Hodgkin's lymphomas. The lymphocyte marker pattern of non-Hodgkin's lymphoma cells was related to current concepts of lymphoma classification. In a series of 28 lymphomas lymphocyte markers indicated that 2 were of histiocytic origin, 2 were unclassifiable, none were derived from T cells and the remainder were B-cell neoplasms. The immunoglobulin heavy chain associated with the B-cell tumours was gamma in one case, alpha in one case but was mu in the majority of cases, reflecting the predominance of this heavy chain, together with delta chains, on normal lymph node lymphocytes in man. delta chains accompanied mu chains on the tumour cells in 6/17 lymphomas in which anti-delta staining was performed. delta chains were not found on any lymphomas other than well differentiated diffuse lymphocytic types. There was evidence of a reduction in surface immunoglobulin, Fcgamma and C3 receptors on undifferentiated lymphoma cells. T lymphocytes of normal morphology were present in all lymphomas except one, and were more numerous in follicular lymphomas than in diffuse tumours.


Characterization of mononuclear cells in the inflammatory infiltrates of cutaneous tumours. The identification of mononuclear cells extracted from cutaneous tumours (basal cell carcinoma, squamous cell carcinoma and superficial, spreading melanoma) has been investigated. The relative numbers of T cells and B cells have been determined using the E-rosette test and the EAC-rosette test. The results have been compared to those of delayed hypersensitivity type reactions. Different cell distribution patterns (E/EAC ratio) have been found in the infiltrates according to the type of tumour. An immunocytochemical technique has been developed for the identification in situ of immunoglobulin-producing cells in the inflammatory infiltrates. In each case the class of immunoglobulin (IgM, IgG or IgA) has been identified and the relative frequency of Ig-producing cells has been determined. The results indicate humoral and cellular immune responses with variations attributable to the type of tumour. In weakly malignant tumours, the infiltrate is characterized by an elevated number of T lymphocytes and numerous plasma cells which secrete all classes of Ig; in highly malignant tumours it is characterized by a reduced number of both T lymphocytes (E rosette) and plasma cells which do not secrete all classes of Ig.


Autologous bone marrow repopulation following high dose cyclophosphamide and allogeneic marrow transplantation in aplastic anaemia. Two patients are described who received allogeneic bone marrow transplantation for aplastic anaemia. Both patients rejected their grafts but subsequently repopulated their marrow with autologous haematopoietic cells. The significance of this phenomenon and its relationship to the possible pathogenesis and therapy of aplastic anaemia and discussed.


Activated T-cell in autoimmune haemolytic anaemia. In 12 of 14 patients with clinically active autoimmune haemolytic anaemia high levels (9-78%) of homologous and autologous rosetting cells were found in the peripheral blood. In 10 of these patients, who were observed over a period of 2 years, the levels bore a direct relationship to the activity of the disease. The cells so identified were T-lymphocytes. The role of cells, identified by this method in AIHA, is discussed.


Chronic lymphatic leukaemia: correlation of immunofluorescent characteristics and clinical features. 60 patients with chronic lymphatic leukaemia (CLL) were classified according to the immunofluorescent characteristics of their lymphocytes. A group expressing surface membrane (Sm) IgM had a significantly less aggressive disease than those expressing a mixture of SmIgM and SmIgD or those without surface immunoglobulin. The patients were older than in most comparable series and had a 2:I preponderance of women. It is suggested that especially among old ladies there is a benign variety of CLL which expresses SmIgMK, and that determination of the SmIg class in CLL has prognostic value.


The detection of granulocyte alloantibodies with an indirect immunofluorescence test. An indirect immunofluorescence technique for the detection of alloantibodies against granulocytes was developed. A major problem was the unspecific fluorescence of granulocytes, due to unspecific adherence of immunoglobulins to the cell membrane. This could be suppressed by the prefixation of the granulocytes with paraformaldehyde. The developed test proved to be specific and sensitive and to have a high reproducibility. Strongly positive reactions were obtained with antisera containing granulocyte-specific agglutinins or granulocyte-cytotoxins, but also with some sera that did not react in either the agglutination or the cytotoxicity test. HLA antisera also gave positive reactions, but strong anti-A, anti-B or anti-D sera did not react. With this technique leucocyte antibodies can be detected in a higher percentage of patients with febrile transfusion reactions.


Effect of bromocriptine on the premenstrual syndrome. A double-blind clinical trial. Twenty-one patients suffering from the premenstrual syndrome were each studied during three menstrual cycles. After a control cycle, bromocriptine and placebo were given during the luteal phase of the cycle in a random double-blind cross-over manner, each patient serving as her own control. The dosage of bromocriptine was 2-5 mg twice daily. Serum prolactin levels were found to equal during the follicular and luteal phases, except when reduced by bromocriptine. Serum progesterone and oestradiol-17-beta were within normal ranges, and did not change during treatment. Medication considerably improved all the premenstrual symptoms, but mastodynia was the only one where bromocriptine was significantly better than the placebo.


Treatment of hypertension in pregnancy with methyldopa: blood pressure control and side effects. A total of 242 women with moderate hypertension in pregnancy completed a controlled trial of methyldopa (Aldomet). The hypotensive effect of methyldopa was similar to its action in non-pregnant individuals and greatly reduced the frequency of severe hypertension occurring antenatally and in labour. As pregnancy advanced, an increasing daily dose of methyldopa was needed and there was a greater use of additional hypotensive therapy. Seventeen (14-5 per cent) women assigned to methyldopa had to be transferred to another drug or had to stop treatment completely because of minor side effects, of which the commonest were lack of energy and dizziness. No serious side effects were encountered. Nine per cent of the untreated women developed severe hypertension which required treatment later in their pregnancies. Six weeks after delivery, nearly all the patients were able to stop treatment.


Treatment of pregnancy sickness.

Treatment of pregnancy sickness. A double-blind comparison was undertaken between Debendox with 10mg of extra pyridoxine and placebo with 10mg of pyridoxine, in 56 women suffering from nausea and/or vomiting during the first 10 weeks of pregnancy. The results of treatment were assessed on the patient's own dialy records of:the time of nausea, the frequency of nausea, and the severity of nausea, retching and vomiting. There were statistically significant differences in favour of Debendox with extra pyridoxine in respect of the days of nausea all day (P les than 0-02), the severity of nausea (P less than 0-05) and the severity of retching (P less than 0.05).


Substrate selectivity of squalene synthetase. Six 1-3H-labeled analogues of farnesyl pyrophosphate have been studied as potential substrates for yeast and rat liver squalene synthetases: 2-methylfarnesyl pyrophosphate (4), 3-demethylfarnesyl pyrophosphate (5), 7,11-dimethyl-3-ethyl-2,6,10-dodecatrienyl pyrophosphate (6), 6,7,10,11-tetrahydrofarnesyl pyrophosphate (7), 4-methylthiofarnesyl pyrophosphate (8), and 4-fluorofarnesyl pyrophosphate (9). Analogues 4 and 5 are enzymatically incorporated into 11-methylsqualene (10) and 10-demethylsqualene (11), respectively, even if no farnesyl pyrophosphate is added to the incubations. None of the other analogues gives nonpolar products with either the yeast or liver enzymes. No tritium is enzymatically released to the medium from any of the analogues, indicating that they are not accepted at the first (proton exchanging) site. The data rule out formation of dead-end presqualene pyrophosphate products with analogues as first, but not as second, substrates. Implications of these results for the enzyme active-site topology and mechanism are discussed.


Adsorption protein of the bacteriophage fd: isolation, molecular properties, and location in the virus. The adsorption (minor coat) protein of the bacteriophage fd has been implicated to function in several steps of viral morphogenesis. The protein has been purified by sodium dodecyl sulfate gel filtration after dissociation of the virus. The adsorption protein preparation was estimated to have less than 5% contamination by analysis on sodium dodecyl sulfate-polyacrylamide gels and by the results of semiquantitative dansyl-Edman degradation. The amino-terminal sequence of the adsorption protein is H2N-Ala-Glx-Thr-Val-Glx-Ser-Pro-Leu-Pro-. Carboxypeptidase A plus B digestion of the protein under a variety of denaturing conditions did not release any amino acids. There are 3-4 adsorption proteins per virion as estimated by the distribution of E114C]leucine between the major and minor coat protein peaks on sodium dodecyl sulfate-polyacrylamide gels. Adsorption protein-specific antibodies were induced in the rabbit and used as electronmicroscopic markers to determine the position of the adsorption proteins in the viral particle. The adsorption proteins were found at only one end of the filamentous viral particles.


Adsorption protein of bacteriophage fl: solubilization in deoxycholate and localization in the fl virion. A complex containing the minor coat protein or adsorptionprotein (A protein) of bacteriophage fl has been solubilized from the fl virion, using the detergent deoxycholate. This complex was resolved from the fl DNA and from the fl major coat protein, or B protein, by gel filtration in the presence of deoxycholate. The A protein complex migrated as a single band on sodium dodecyl sulfate-urea-polyacrylamide gels corresponding to a molecular weight of 60 000. Analysis of the amino acid composition and amino terminal residues of this preparation indicates that the preparation contains a 20% contamination of additional protein species. Antibody against purified fd A protein is cross-reactive with deoxycholate-purified fl A protein and with fl phage. Electron microscopic observation of negatively stained complexes of fl phage with this anti-fd A protein antibody and ferritin conjugated goat anti-rabbit IgG antibody revealed phages with ferritin particles at their termini or complexes of two or more phages joined together at one end by ferritin, indicating that the complex of A protein molecules is located at one end of the filamentous fl virion.


Thermodynamic studies of the reversible association of Escherichia coli ribosomal subunits. The kinetics of association of Escherichia coli 30S and 50S ribosomal subunits have been carried out as a function of temperature after a magnesium jump from 1.5 to 3 mM. Turbidimetric recordings combined with a stopped-flow apparatus were used to follow the kinetics. The data show that the rates of formation and dissociation of the 70S particles at 3 mM Mg2+ and +25 degrees C were, respectively: k2 = 10(5) M-1 s-1, k1 = 4,5 X 10(-3) s-1; lowering the temperature decreases the rate constants with activation energies equal to E2 = 7.5 kcal/mol, E1 = 26.5 kcal/mol and enhances the association equilibrium towards the 70S species with an enthalpy change (delta H degrees assoc = -19.9 kcal/mol) dominant over the entropy change (delta S degrees assoc = -33 cal/(deg mol)). These thermodynamic parameters were compared to those obtained from studies on the interactions of codon-anticodon in yeast phenylalanine transfer RNA as well as of ribooligonucleotides. The kinetic and thermodynamic data are shown to be consistent with 16S-23S RNA interaction.


Subunit topography of RNA polymerase from Escherichia coli. A cross-linking study with bifunctional reagents. The quaternary structures of Escherichia coli DNA-dependent RNA polymerase holenzyme (alpha 2 beta beta' sigma) and core enzyme (alpha 2 beta beta') have been investigated by chemical cross-linking with a cleavable bifunctional reagent, methyl 4-mercaptobutyrimidate, and noncleavable reagents, dimethyl suberimidate and N,N'-(1,4-phenylene)bismaleimide. A model of the subunit organization deduced from cross-linked subunit neighbors identified by dodecyl sulfate-polyacrylamide gel electrophoresis indicates that the large beta and beta' subunits constitute the backbone of both core and holoenzyme, while sigma and two alpha subunits interact with this structure along the contact domain of beta and beta' subunits. In holoenzyme, sigma subunit is in the vicinity of at least one alpha subunit. The two alpha subunits are close to each other in holoenzyme, core enzyme, and the isolated alpha 2 beta complex. Cross-linking of the "premature" core and holoenzyme intermediates in the in vitro reconstitution of active enzyme from isolated subunits suggests that these species are composed of subunit complexes of molecular weight lower than that of native core and holoenzyme, respectively. The structural information obtained for RNA polymerase and its subcomplexes has important implications for the enzyme-promoter recognition as well as the mechanism of subunit assembly of the enzyme.


Ficellomycin and feldamycin; inhibitors of bacterial semiconservative DNA replication. The two peptide-like antibiotics ficellomycin and feldamycin impair semiconservative DNA replication but not DNA repair synthesis in bacteria. Specifically both antibiotics cause the accumulation of a 34S DNA species in toluenized Escherichia coli cells which lacks the capability of being integrated into larger DNA pieces and eventually the complete bacterial chromosome. Novobiocin, a known inhibitor of replicative DNA synthesis, was investigated for comparative purposes. The action of this latter antibiotic differs from the ones exerted by ficellomycin and feldamycin in the novobiocin appears to block an event associated with the initiation of Okazaki fragments. The fact that novobiocin impairs DNA gyrase suggests that this enzyme plays an essential role during the initiation of Okazaki pieces.


Stereochemistry of internucleotidic bond formation by tRNA nucleotidyltransferase from baker's yeast. Isomer A of adenosine 5'-O-(1-thiotriphosphate) (ATP alpha S) is a substrate for tRNA nucleotidyltransferase from baker's yeast, whereas isomer B is a competitive inhibitor. The tRNA resulting from this reaction has a phosphorothioate instead of a phosphate diester linkage at the last internucleotidic linkage between cytidine and adenosine. On limited digestion of this tRNA with RNase A, one can isolate cytidine 2',3'-cyclic phosphorothioate which can be deaminated to uridine 2',3'-cyclic phosphorothioate. It can be shown that this compound is the endo isomer and that, therefore, the phosphorothioate diester bond in the tRNA must have had the R configuration. This result indicates that no racemization during the condensation of ATP alpha S, isomer A, onto the tRNA had occurred. Whether inversion or retention of configuration had taken place awaits elucidation of the absolute configuration of isomer A of ATP alpha S.


Effect of proteolysis of transcriptional fidelity of reconstituted chromatin. The effect of proteolysis on the transcriptional properties of reconstituted rat liver chromatin was studied. Within the sensitivity of currently available methods, proteolysis of chromosomal proteins by chromatin-bound protease during chromatin reconstitution has no apparent effect on: (1) number of initiation sites, (2) proportion of reiterating and unique sequences of DNA transcribed, (3) size of the RNA transcribed, and (4) transcription of DNA sequences complementary to poly(A) containing messenger RNA.


Incorporation of phosphorothioate groups into fd and phi X174 DNA. We have synthesized fd and phi X174DNA in the presence of 2'-deoxyadenosine 5'-O-(1-thiotriphosphate) (dATP alpha S) and the corresponding phosphorothioate derivatives of dCTP and dTTP using ether-permeabilized E. coli cells or crude cell extracts of E. coli DNA polymerase I. Reaction rates of enzymes involved in the formation or breakdown of DNA are decreased in the presence of phosphorothioates. The amount of label incorporated with [35S]dATP alpha S suggests that the dAMP has been completely substituted by 2'-deoxyadenosine 5'-0-phosphorothioate (dAMPS). The substituted DNAs have the same sedimentation coefficients, similar buoyant density, infectivity, and thermal stability as the unsubstituted DNAs. The procedure therefore allows specific modification at the 5' position of dA, dC, or dT in the DNA. In view of the recent demonstration of specific binding of Pt2+ complexes to the phosphorothioate analogue of poly[r(A-U)] (Strothkamp, K.G., and Lippard, S.J. (1976), Proc. Natl. Acad. Sci. U.S.A. 73, 2536), the synthesis of phosphorothioate containing DNA may be of use for DNA sequencing by electron microscopy.


Discrimination between Rb+ and K+ by Escherichia coli. 1. The K+ requirment of Escherichia coli is only partially fulfilled by Rb+. The molar growth yield on Rb+ was about 5% of that on K+ and the growth rate in Rb+-supplemented media is lower thatn in K+ influx by any of the four K+ transport systems of E. coli. The high-affinity Kdp system (Km = 2 micron) is poorly traced by 86Rb+. It discriminates against a 86Rb+ tracer at least 1000-fold. The two moderate affinity systems, the high-rate TrkA system (Km = 1.5 mM) and the moderate rate TrkD system (Km = 0.5 mM), discriminate against a 86Rb+ tracer by approximately 10-fold and 25-fold, respectively. 86Rb+ is preferred by the low-rate TrkF system and overestimates its K+ influx by 40%.


Lipid-protein interactions in the plasma lipoproteins. The purpose of this review has been to discuss new information about the mechanism of lipid and apoprotein interaction in the plasma lipoproteins. A special form of the amphipathic helix has been identified as a major structural element of the apolipoproteins sequenced to date. Evidence is reviewed concerning the role of the amphipathic helix in the binding to phospholipids. Several different models for the organization of the components of HDL, LDL and LP-X have evolved from extensive structural studies. Resolution of the differences among these models will require additional experimental testing. Verification of models based on the study of reconstituted HDL will require rigorous proof of native structure in these particles. A detailed description of the molecular organization of the lipid and protein constituents of the plasma lipoproteins is still lacking. Further structural and sequence studies with apoB and the "arginine-rich" protein are needed. Crystallization of an apoprotein or lipoprotein and determination of the three-dimensional structure would be a major achievement. With such further detailed structural information, it may then be possible to correlate changes in structure with determinants of metabolism.


Translation in Escherichia coli mini-cells containing hamster mitochondrial DNA-Co1E1 - Ampr recombinant plasmids. Hamster mitochondrial DNA is cleaved into two fragments (4.2 and 11.4 kilobase pairs of DNA (kb)) by the restriction enzyme, Eco RI. Recombinant DNA molecules formed in vitro between an Escherichia coli plasmid, Co1E1 - Ampr, and Eco RI-digested hamster mitochondrial DNA were transformed into E. coli K12. The translation products of the parent plasmid, Co1E1 - Ampr, and recombinant plasmid DNAs containing (i) the 4.2 kb mitochondrial DNA fragment and (ii) the 11.4 kb fragment were characterized on sodium dodecyl sulfate-polyacrylamide gels using bacterial mini-cell lysates. The Co1E1 - Ampr plasmid specifies at least six polypeptides whose structural genes comprise 56% of the plasmid DNA. Insertion of hamster mitochondrial DNA at the Eco RI site of the plasmid alters the relative rate of synthesis of these six polypeptides and induces the occurrence of a new band on sodium dodecyl sulfate-polyacrylamide gels which is probably not specified by the inserted mitochondrial DNA sequences.


Unidirectional replication of the P-group plasmid RK2. The mode of replication of the broad host-range plasmid RK2 has been determined from examination of molecular replicative forms cleaved with the restriction endonucleases EcoRI and Hind III. Replication is unidirectional, and proceeds from a unique origin. The location of the origin and other evidence suggests that genes involved in plasmid maintenance are not tightly clustered.


Lutropin stimulation of RNA synthesis in corpus luteum chromatin. Lutropin and human choriogonadotropin stimulated the endogenous chromatin-associated polymerase activity in purified chromatin prepared from nuclei of bovine corpus luteum. Chromatin was incubated in two different buffer systems: one that mainly supports the activity of polymerase I, another that supports the activity of polymerase II and is largely alpha-amanitin sensitive. The hormones lutropin and chorigonadotropin stimulated an increase in the rate of incorporation of [14C]ATP or [14C]UTP into RNA in both buffer systems. Follitropin, prolactin and beta-corticotropin had no stimulatory effect. Neither the alpha nor beta subunit of lutropin stimulated RNA synthesis. When premixed, the subunits rapidly formed the active molecule. A maximum response to RNA synthesis was achieved by a 10(-9) M concentration of human choriogonadotropin. Considerable activity was obtained at 10(-11) M human choriogonadotropin. There was no lutropin stimulation to RNA synthesis using calf thymus DNA and Escherichia coli RNA polymerase.


Expression of a bacterial gene for guanine synthesis inserted into colicin E1 factor by an in vitro recombination. Regulation of expression of a bacterial guaA gene inserted into colicin E1 DNA by an in vitro recombination was studied under various growth conditions. In Escherichia coli K-12 cells that carried this hybrid ColEl plasmid the level of guaA enzyme activity was not regulated by the concentration of guanine in the medium, but by the number of plasmid DNA copies. The optimal conditions for amplifying the guaA gene product by chloramphenicol treatment were determined. The level of guaA enzyme activity found under the optimal conditions was about 37 times that in extracts of wild-type E. coli cultured in guanine-free medium. The properties of the promoter for the guaA gene and applicability of this hybrid ColEl plasmid for amplification of various gene products were discussed.


Thermometric enzyme linked immunosorbent assay: TELISA. A new method, thermometric enzyme linked immunosorbent assay (TELISA), for the assay of endogenous and exogenous compounds in biological fluids is described. It is based on the previously described enzyme linked immunosorbent assay technique, ELISA, but utilizes enzymic heat formation which is measured in an enzyme thermistor unit. In the model system studied determination of human serum albumin down to a concentration of 10(-10) M (5 ng/ml) was achieved, with both normal and catalase labelled human serum albumin competing for the binding sites on the immunosorbent, which was rabbit antihuman serum albumin immobilized onto Sepharose CL-4B.


Modification of tyrosine residues of the lactose repressor protein. Reaction of the lactose repressor protein from Escherichia coli with high molar excesses (up to 800 fold) of tetranitromethane resulted in modification of tyrosine residues in the amino-terminal and core regions of the molecule. Tyrosines 7 and 17 exhibit significant reactivity at low levels (5-10 fold molar excess) of tetranitromethane. The loss of operator binding activity upon nitration at these low concentrations of reagent indicates involvement of these two tyrosines in the binding process. Inducer binding activity was maintained at approx. 90% of unreacted repressor for all excesses of reagent studied. Addition of inducer to the repressor prior to reaction resulted in decreased modification of tyrosines in the core region, but anti-inducers did not affect the reaction significantly. The effect of inducers on the pattern of reaction apparently reflects the conformational change which occurs upon binding of these ligands. Acetylation of the repressor protein with N-acetylimidazole modified lysines and tyrosines with complete loss of operator binding activity and retention of 75-80% of inducer binding activity.


Histidinol dehydrogenase from salmonella typhimurium and Escherichia coli. Purification, some characteristics and the amino acid sequence around a reactive thiol group. The purification and some physical properties of histidinol dehydrogenase, L-histidinol-nicotinamide adenine dinucleotide oxido-reductase (EC 1.1.1.23) from either Salmonella typhimurium or Escherichia coli are reported in this paper. Modification of histidinol dehydrogenase with one equivalent of N-(4-dimethylamino-3,5-dinitrophenyl)maleimide at pH 6.8 yields an enzyme that is inactive toward the oxidation of L-histidinol. The modified cysteine residue was located in an acid insoluble tryptic core. The amino acid sequence around the reactive thiol group in S. typhimurium is: Leu-Cys-Gly-Val-Glu-Glu-Ile-Phe, and in E. coli is: Leu-Cys-Gly-Val-Glu-Asp-Val-Phe. These unique sequences show no homology to the reactive thiol groups from some other dehydrogenases.


The yeast aminoacyl-tRNA synthetases. Methodology for their complete or partial purification and comparison of their relative activities under various extraction conditions. Several fractionation steps are described which can be applied to the partial purification of the 20 aminoacyl-tRNA synthetases from commercial baker's yeast. Comparative experiments performed in the presence or absence of protease inhibitors revealed that some enzymes prepared in the presence of the inhibitor exhibit much higher specific activities than the proteins extracted in the absence of the inhibitor. The methodology reported can be used for the simultaneous preparation of several pure aminoacyl-tRNA synthetases. As examples, the large scale purification of phenylalanyl-and valyl-tRNA synthetases are described.


[On the role of N7 atoms of guanosine in tRNA-Phe (E. coli) in interaction with ribosomes]. The influence of alkylation of phe-tRNAPhe (E. coli) with 2',3'-O-[4-(N-2-chloroethyl-N-methylamino)-benzylidene]-uridine - 5' - methylphosphate on its ability to participate in non-enzymatic complex formation with ribosomes and poly-U was investigated. Phe-tRNAsPhe, containing alkylated guanosines at different positions, including anticodone, are active in binding with ribosomes. It is concluded, that N7 nitrogens of guanosine of the tRHAPhe are not elements, significant for the interaction with ribosomes.


[Some peculiarities of phage DDVI-specific methylases]. The types of methylases are found in the cellular extract of Escherichia coli B, infected with phage DDVI. One of them is a cellular enzyme, which methylates adenine to form 6-methylaminopurine (6-MAP) and is repressed in the infected cell in vivo. The second type, which is not found in the non-infected cells, is specific for phage DDVI and induces the formation of 7-methylguanine (7-MG). Both enzymes recognize various sites, which accounts for the ratio 6-MAP/7-MG to vary in heterological DNAs between 2.07 in phage Sd DNA and 0.40 in phage DDII DNA. During in vitro incubation with homologous methylases phage DDVI DNA and especially phage T2 DNA are subjected to further methylation, which is probably indicative of their "undermethylation" in vivo. The DDVI-specific enzyme, similar to B-specific type, methylates DNA with a normal set of nitrogenous bases (phages Sd and DDII), as well as DNAs containing 5-oxymethylcytosine and glucose (phages T2 and DDVI). Both methylases under study use only native double-helical DNA as substrate and are strongly inhibited by S-adenosylhomocysteine. Phage DDVI Methylase is characterized by low stability.


[Cinematics of fibroblast spreading. I. Analysis of a population]. Normal mouse fibroblasts were studied by means of time-lapse microcinematography under phase-contrast microscope. For 31 cells the area measurements during spreading were made with the intervals of 2.5--5 minutes. The following results have been obtained. The maximal area of spreaded cell is 3--4 times greater than the surface area of the sphere with the spherical cell radius. There is a tendency of the cell area to increase linearly in the middle, that is in a regime of a constant middle rate. Most cells spread either in one regime, or in two, when a more rapid regime is followed by a shower one. The change of the spreading regime is a result of the control of mean rate of spreading.


[Limited proteolysis of bovine pepsin].

[Limited proteolysis of bovine pepsin]. A limited proteolysis of bovine pepsin (EC 3.4.4.1) was carried out. A proteolysis-resistant C-terminal protein fragment containing about 170 amino acid residues was isolated and its N-terminal sequence was established, using Edman's automatic method. It was assumed that the fragment of bovine pepsin isolated, similar to the previosly obtained porcine pepsin fragment, is an independent constituent of the protein molecule.


[T2 DNA, modified by 2,2,6,6-tetramethyl-4-bromoacetooxypiperidine-i-oxyl as a template for RNA polymerase from E. coli B]. T2-DNA was modified by 2,2,6,6-tetramethyl-4-bromoacetooxypiperidine-1-oxyl (I) at different NaCl concentrations (10(-1) M NaCl--10(-4) M NaCl). Modified DNA were investigated as templates for the RNA-polymerase from E. coli B. It was shown that T2-DNA modified I in 0,1 M NaCl completely preserves the native secondary structure, has a low degree modification (1 molecule I per 1000-2000 nucleotide pairs), but is a noneffective template for the RNA-polymerase from E. coli B (20%-40% as compared with unmodified T2-DNA). Under these conditions the modification occurs probably at the "weakest" (readily melting) sites of DNA. The role of these "weak" sites on DNA as promotors is discussed. The modification of T2-DNA by reagnet I has a stronger inhibitory effect on the total RNA synthesis than on the RNA-synthesis stable to rifampicin. Possible existence of two kinds of "early" promotors on T2-DNA is assumed.


[Proton translocation in membranes of submitochondrial particles]. Effect of an electrophilous inhibitor, chlorophenacyl, on energy-dependent functions of submitochondrial particles is studied. Chlorophenacyl at concentrations up to 1 mM is found practically not to affect the generation of membrane potential under NADH and succinate oxidation and ATP hydrolysis and to be a strong inhibitor of oxidative phosphorylation and reverse electron transport. The mechanism of the inhibition of energy-dependent functions of submitochondrial particles with chlorophenacyl is different from that of electron transport inhibitor, energy transport inhibitors and classical uncoupling agents--protonophors. The data obtained are suggested to be due to the existence of two ways of proton translocation in submitochondrial particle membrane, phosphorylating and non-phosphorylating, the effect of chlorophenacyl being directed on phosphorylating way only.


Microbiological relevance and clinical potential of ampicillin-cloxacillin synergism. Recent demonstration "in vitro" that combinations of ampicillin and cloxacillin, using concentrations at which neither were previously effective, will kill certain resistant Gram negative bacteria, has important clinical potential. We therefore studied 50 ampicillin resistant Gram negative rods cultured from septicemic patients for synergism. 7 of 23 E. coli strains with a minimal bactericidal concentration (MBC) of 128 microgram/ml were killed by combinations containing 4-32 microgram/ml ampicillin and 16-32 microgram/ml cloxacillin. These same strains were also resistant to cephalothin. Synergism was also noted in 1/4 strains of Enterobacter, 1/3 strains of Serratia, 5/14 strains of Klebsiella and 1/6 strains of Pseudomonas. In separate experiments ampicillin (56 mg/kg) and cloxacillin (14 mg/kg) was administered intravenously over a period of thirty minutes. Peak serum levels measured simultaneously at the end of the infusion were 235 +/- 11 (SEM) microgram/ml for ampicillin and 85 +/- 7 microgram/ml for cloxacillin. Levels of ampicillin/cloxacillin measured one and two hours after the infusion were 117/32 and 43/8 microgram/ml. T 1/2 of ampicillin and cloxacillin were 0.8 and 0.5 h respectively. Since the serum concentrations of both antibiotics measured one hour after the infusion were greater that levels required for "in vitro" synergism, we suggest that combinations of ampicillin and cloxacillin would be of importance in the treatment of Gram negative septicemia, since such therapy would increase the bactericidal effect of the sera without increasing the risk of toxic side effects.


Collagen stimulates more platelet prostaglandin endoperoxides in thrombosis. Platelets prostaglandin cyclic endoperoxides after collagen stimulation were measured in 7 patients with arterial thrombosis, 4 patients with recurrent deep venous thrombosis and 27 patients before undergoing total hip replacement. Patients with post-surgical D.V.T. synthetized more platelet cyclic-endoperoxides compared to those with no D.V.T. COULD BE DIVIDED INTO 2 SUBGROUPS : GROUP I with platelet cyclic endoperoxides and group II with higher cyclic-endoperoxides synthesis. Also, there was a higher ratio of platelet cyclic endoperoxides in patients with arterial thrombosis and in 2 patients with recurrent venous thrombosis and shortened platelet survival time. Thus, a greater synthesis of platelet prostaglandin cyclic, endoperoxides was formed in a number of patients with thrombosis.


Immunologic status of cancer patients : correlation between number of active rosettes and analytical electrophoresis of lymphocytes. Clinical findings in cancer patients and experimental results after separation of active rosettes-forming cells are compatible with the hypothesis that the lymphocytes which do form active rosettes are those whose electrophoretic mobility is about 1.20 micron. sec-1v.-1.cm.


Human autologous and allogeneic rosettes with erythrocytes of the Bombay type. Human red blood cells of the Bombay type which lack ABH group substances can bind to allogeneic lymphocytes just as well as erythrocytes of any other type. A much lower percentage of auto-rosettes between erythrocytes and lymphocytes from the Bombay donor was observed, a result which may be due at least partially to some T lymphocyte defect in the Bombay donor.


An approach to simultaneous immunofluorescent and autoradiographic analysis of human normal and neoplastic blood cells. A technique for evaluating membrane immunofluorescence and isotope (3H-Thymidine, 3H-Uridine, 3H-Leucine) labelling in the same cell is described in detail. The possible interference of autoradiographic labelling on the fluorescent staining have been considered and found not to alter the final preparations. The technique proved valid both in normal and in neoplastic cells irrespective of their origin (peripheral blood, bone marrow, lymph node). Its possible extensions are finally discussed.


A phase II clinical trial of prednimustine. Clinical screening cooperative group of E.O.R.T.C. In a phase II trial, prednimustine was often efficient in treating chronic lymphoid leukaemia (CLL) patients and was also active in some patients with lymphosarcoma, melanoma and bronchus carcinoma. Tolerance was generally excellent, the most critical side effect being thrombocytopenia in the case of CLL.


A kinetic model of cooperativity in aspartate transcarbamylase. A relatively simple kinetic model is proposed to account simultaneously for data on the binding of carbamyl phosphate and succinate to aspartate trans carbamylase (ATCase), and for the relaxation spectrum associated with this binding. The model also accounts for measurements of the initial velocity of the reaction of ATCase with respect to aspartate and carbamyl phosphate. The principal assumption made is that ATCase consists of three identical noninteracting cooperative dimers. Ordered binding and both sequential and concerted conformational changes in the dimers are needed to account for the properties of ATCase. The values of the parameters of this model can be determined by fitting to existing experimental evidence. Various new quantitative predictions are made that can serve as additional tests of the proposed theory.


Some observations on oscillatory changes in the growth rate of Saccharomyces cerevisiae in aerobic continuous undisturbed culture. Oscillatory changes in the growth rate were observed in undisturbed continuous culture of Saccharomyces cerevisiae on sugar-cane molasses media when nitrogen sources (2.56 to 6.17 g/liter of ammonium sulfate or 1.22 g/liter of urea) were added to the feeding mash and when the air rate was 1.3 to 1.6 v/v/m. The oscillations were not affected by the addition of yeast extract. The suppression of the nitrogen source during the continous test leads to a nonoscillatory transient state. No oscillations occured at all when no nitrogen source was added to the medium and/or the air rate was equal to zero or equal to about 3.3 v/v/m. The oscillatory responses of the system were affected by a previous anaerobic continuous cultivation of the yeast.


[Test-strains of E. coli for detection of chemical mutagens]. Two temperature-sensitive mutants--AP 16 and AP 18 were isolated after the treatment of E. coli AB2500 strain with two mutagens (acridine orange and 5-bromuracil). The mutants obtained proved to be sensitive and formed revertants when treated with the following agents: N-methyl-N'-nitro-N-nitrosoguanidine, hydroxylamine, nitrous acid, sodium metabisulfite, methylmethansulfonate, and proflavine. Introduction into the mentioned strains of additional mutation causing elevation of their sensitivity to crystal violet increased somewhat their capacity to form revertants under the effect of proflavine and methylmethansulfonate.


[Transformation of the substrate specificity of EcoRI restrictase under the influence of glycerin]. The restriction endonuclease EcoRI hydrolyzes DNA to a greater number of fragments in the presence of glycerol than under normal conditions. This enzyme begins to work by the so-called EcoRI-type of restriction when glycerol concentration reaches 50%. The EcoRI activity appeared in experiments only when the ionic strength of the solution was decreased and pH of the solution was increased. However, under such extreme conditions the enzyme was quickly inactivated and it was difficult to obtain reproducible results especially for hydrolysis of the high-molecular DNA. The suggested conditions for the EcoRI activity permit to obtain reproducible results, this being practically equivalent to discovery of the new restriction endonuclease.


[Cytopathogenic effect of E. coli cells containing heterogeneous human O(H) type antigen on human cells in culture]. Interaction of two enteropathogenic strains E. coli O55-K59 and human Hela cells containing O(H) isoantigen was studied. When E. coli strain No. 5789 containing heterologic antigen O(H) was added to HeLa cell culture the cytopathogenic effect with the microbial doses of 2 X 10(10), 2 X 10(5), 2X 10(4) was revealed on the third day of the interaction. A dose of 2 X 10(3) of E. coli microbes gave no such effect. Strain No. 3827 containing no heterologic antigen of ABO type failed to exert any cytopathogenic effect with maximal, mean, and minimal doses of the microbes. It is assumed that the cytopathogenic effect of strain No. 5789 is connected with the presence of the strain antigen identical to the group antigen of the human cell culture under study.


[Specificity of antinuclear autoantibodies in diseases with systemic destruction of connective tissue]. By using the fluorescent antibody technique several patterns of antinuclear antibodies (ANA) were detected in the systemic connective tissue diseases, as well as in some other diseases and in healthy donors, depending on the character of nuclear fluorescence. Homogeneous nuclear fluorescence, homogeneous fluorescence free of nucleaolar fluorescence, ring-shaped fluorescence, lumpy fluorescence, selective nuclealar fluorescence, nuclear fluorescence in the form of long fine plexiform bands associated with fluorescence in the nuclear membrane region. The latter ANA pattern was found only in several forms of lupus erythematosus, and this fluorescence was different from the previously reported "reticular" and "filamentous" patterns.


[Permeability of cutaneous macrophage lysosomal membrane in delayed hypersensitivity to streptococcal antigens]. The acid phosphatase (APh) was studied in the skin macrophage lysosomes of animals with delayed-type hypersensitivity (DTH) to the antigens of group A streptococci or tuberculoproteins. The histochemical procedure for the APh determination was employed in the skin sections without any preliminary tissue fixation. Intradermal injection of specific antigens in DTH proved to lead to the increase of the lysosomal membrane permeability in the skin macrophages. These data support the supposition that the lysosome enzymes of macrophages can act as factors causing tissue destruction in DTH.


Skin cover for exposed cardiac pacemakers. Skin flap cover of exposed cardiac pacemakers in 3 patients was successful in 2. In the third, repeated plastic surgical procedures enabled the patient to have the continuous use of his pacemaker for nearly a year.


Anaphylactic reactions to endotoxin in guinea-pig tissues: relationship to endotoxin toxicity. 1 A lipopolysaccharide extract of Escherichia coli 026:B6 cells (026:B6(B) endotoxin) was shown to be toxic to normal adult guinea-pigs. 2 The agent had no action on isolated preparations of ileum and heart taken from normal adult guinea-pigs. 3 Ileal segments from animals actively immunized against 026:B6(B) endotoxin showed dose-dependent contractions when exposed to endotoxin. Desensitization phenomena were demonstrated. 4 Reactivity of 026:B6(B) endotoxin was transferred to isolated preparations of ileum and heart from normal animals by passive transfer of immune serum. 5 Tissue responses to 026:B6(B) were associated with release of ileal spasmogen into the bath medium. Mepyramine blocked the effects of this spasmogen at bath concentrations which caused little change in ileal responses to carbachol. 6 It is concluded that E. coli endotoxin can elicit anaphylactic reactions, and that this process may potentiate endotoxin toxicity in sensitized animals. However, endotoxin toxicity in guinea-pigs does not appear to depend on this kind of allergic process.


Feline endotoxin shock: effects of methylprednisolone on kininogen-depletion, on the pulmonary circulation and on survival. 1 Escherichia coli endotoxin, administered intravenously in a dose of 2 mg/kg to pentobarbitone anaesthetized, artificially ventilated cats resulted in pulmonary hypertension, systemic hypotension and an immediate (1-2 min) 30-40% reduction in plasma kininogen, an effect which probably indicates a release of plasma kinins. 2 Methylprednisolone (30 mg/kg), when administered 30 min before endotoxin, did not influence the endotoxin-induced pulmonary hypertension or systemic hypotension but completely prevented the depletion of plasma kininogen. 3 In spontaneously breathing cats, methylprednisolone, administered 30 min after endotoxin, caused a rapid repletion of kininogen and prolonged survival (47% at 6 h compared to 10% in the endotoxinalone animals). Methylprednisolone did not appear to influence lactate production or the hyperventilation observed during the delayed endotoxin shock phase. 4 It is concluded t,at methylprednisolone does not prevent the release, by  endotoxin, of a pulmonary vasoconstrictor prostaglandin, or its effects, but that perhaps by preventing kinin release it may reduce endotoxin-induced capillary leakage.


Mechanical suture methods in thoracic and abdominal surgery. Mechanical suturing procedures on the bowel, lung, bronchi and pulmonary vessels have been used on 264 occasions in 180 thoracic and abdominal operations. The instruments proved simple to use and produced reliable good quality closures, for the most part without manual reinforcement. There were 2 technical failures, one ending fatally, after colo-colostomy. Two minor postoperative suture leaks were encountered in 122 operations on the alimentary tract, but no leakage or other problem has arisen in 58 pulmonary cases. The staple-suturing methods described are safe and straightforward, and it is believed that they represent a significant advance in the techniques of thoracic and abdominal surgery, which can usefully be applied in many situations.


Computer-aided diagnosis of lower abdominal pain in women. This paper describes the use of a system of computer aided diagnosis in an unselected, prospective survey of 393 women suffering from lower abdominal pain of less than 1 week's duration. An accurate diagnosis was made by clinicians at first patient contact in 68-5 per cent of the group of patients. The computer's diagnostic prediction (based on the same data) matched the final diagnosis in 81-6 per cent of the patients. During this survey a marked improvement in diagnostic accuracy was observed amongst the junior clinicians. It is suggested that this is because of the discipline of data collection imposed and the intermittent feedback received, and also that this educational aspect of computer usage may be of wider benefit.


Renal autotransplantation in the treatment of hypertension and uraemia due to renal artery stenosis. Four cases of renovascular hypertension cured or improved by renal autotransplantation are described. In one case correction of renal ischaemia resulted in an improvement of renal function. Previous reports of this technique are reviewed and the limitations of the more standard operation of saphenous vein bypass graft are discussed.


Abdominal wound closure: a trial of nylon, polyglycolic acid and steel sutures. Failures after abdominal wound closure (early dehiscences and late incisional hernias) are due to breakage of sutures, slippage of knots or tearing out of sutures from the tissues. The suture-holding capacity of the entire thickness of muscle and aponeurosis is nearly twice that of the anterior rectus sheath, and deep bites (1-0 cm from the cut edges) are nearly twice as secure as bites of 0-5 cm. In a random controlled clinical trial of 357 major laparotomies, closure with either layered monofilament nylon or mass polyglycolic acid or steel resulted in 2 burst abdomens (0-56%), 10 incisional hernias due to suture failure (3-4%) and 8 incisional hernias caused by deep sepsis (2-7%). There were no statistically significant differences among the treatment groups, but 1 patient in the nylon and 3 in the steel groups had persistent sinuses until their sutures were removed.


Abdominal wound healing: a prospective clinical study. The incidence of wound dehiscence and incisional hernia after two methods of abdominal wound closure (layered closure with retention sutures and single-layer "mass closure") was studied in a randomised prospective clinical trial in a consecutive series of 200 patients. Dehiscence occurred in 1% of patients and herniation in 4-7%; the incidence of these complications was similar with both methods of closure. Seven of the 11 cases of dehiscence or herniation occurred in infected wounds, and wound infection was associated with a tenfold increase in the incidence of these complications. The prevention of wound infection would reduce substantially the incidence of dehiscence and herniation in abdominal wounds.


Spina bifida and anencephaly: are miscarriages a possible cause? The marriage-to-conception interval in 151 pregnancies producing infants with anencephaly or spina bifida was not significantly different from that in 218 pregnancies resulting in normal infants. Significantly more miscarriages occurred before than after the birth of 285 infants with anencephaly or spina bifida, but in 133 controls no before-after difference was observed. These observations seem to favour the idea that miscarriages are a manifestation rather than a cause of anencephaly and spina bifida.


Reduction in mortality after myocardial infarction with long-term beta-adrenoceptor blockade. Multicentre international study: supplementary report. In a controlled multicentre trial carried out to assess the value of long-term practolol treatment after myocardial infarction the provisional results showed a significant reduction in mortality, though some of the data were lacking. These have now been included and the results updated. The final figures for all deaths were 78 in the placebo group of 1533 patients and 48 in the practolol group of 1520 patients. The reduction in mortality (38%) was significant at the 1% level. The figures for non-fatal reinfarction (97 in the placebo group, and 75 in the practolol group) were not significantly different. Patients with pre-entry anterior infarction, and especially those with a diastolic blood pressure equal to or below the mean (78 mm Hg) at entry to the trial, were at high risk but benefited particularly well from beta-adrenoceptor blockade. After pre-entry inferior infarction the percentage reduction in deaths occurring within two hours after symptoms of a new event was similar to that after anterior infarction, but the incidence of death more than two hours after the event was greater in the practolol-treated group. Thus the difference between groups in total deaths after pretrial inferior infarction was marginal. Until the results of further trials are reported long-term beta-adrenoceptor blockade (possibly up to two years) is recommended after uncomplicated anterior myocardial infarction.


Cellular reaction to group A beta-haemolytic streptococcal membrane antigen and its relation to complement levels in patients with rheumatic heart disease. Cell-mediated immunity and blood complement activities were studied in 35 patients with chronic rheumatic heart disease (RHD) and 17 normal subjects. The T-cell population in patients with RHD was reduced, as were the CH50 and C3 complement levels. The response to phytohaemagglutinin stimulation was deficient, but the lymphocytes of patients with RHD showed increased avidity for 3H-thymidine when stimulated with specific streptococcal membrane antigen. No differences were found between patients with acute rheumatic activity and those without such activity. The susceptibility of individual patients may be related to the specific sensitisation of lymphocytes, while the fact that this persisted even when T-cell numbers had returned to normal may account for the well-known recrudescenses after streptococcal infections in these patients.


Treatment of acute lymphoblastic leukaemia: effect of variation in length of treatment on duration of remission. Report to the Medical Research Council by the Working Party on Leukaemia in Childhood. Patients with acute lymphoblastic leukaemia (ALL) were allocated at random either to stop maintenance chemotherapy after six 12-week courses or continue with a further six. The main difference between the two groups was in the incidence of bone-marrow relapse within nine months after stopping treatment. Such relapses occurred less in older patients and those with higher leucocyte counts initially than in those who appeared to have the best prognosis--namely, those with typical low-count childhood ALL. No patient given prophylactic irradiation to cranium and spine combined with intrathecal methotrexate suffered meningeal relapses, whereas among those not given such prophylaxis the lack of benefit from continuing treatment was mainly attributable to meningeal relapses.


Contribution of atenolol, bendrofluazide, and hydrallazine to management of severe hypertension. The efficacy of various combinations of atenolol, bendrofluazide, and hydraliazine given twice daily was assessed in a double-blind trial on 39 patients with moderate to severe essential hypertension. Concurrent treatment with all three drugs proved most effective and produced a mean reduction in blood pressure of 43/31 mm Hg. In the dosage used, hydrallazine affected only the diastolic blood pressure, and when added to either bendrofluazide or bendrofluazide plus atenolol it produced a further mean reduction in pressure of 6 mm Hg. Once-daily treatment with atenolol and bendrofluazide was as effective in reducing blood pressure as the same combination given twice daily, and the hypotensive effect was still present at least 24 hours after the last dose of tablets. A combined tablet of atenolol and bendrofluazide taken once daily would be a simple regimen to follow and would provide almost as much hypotensive effect as a twice-daily regimen incorporating a modest dose of hydrallazine. The hypotensive effect of atenolol was equal to that of bendrofluazide on systolic pressure but significantly better than that of bendrofluazide on diastolic pressure. Atenolol reduced plasma renin and urate concentrations but increased plasma potassium levels. The biochemical effects of atenolol, therefore, may be an advantage over those of bendrofluazide when deciding on first-line treatment for essential hypertension.


Amoxycillin and co-trimoxazole in presumed viral respiratory infections of childhood: placebo-controlled trial. A double-blind randomized controlled trial of amoxycillin, co-trimoxazole, and placebo was conducted on 197 children presenting with presumed viral respiratory infections. Routine throat swabs were taken to exclude streptococcal diseases. The three disease categories studied--nasopharyngitis, pharyngotonsillitis, and bronchitis (including laryngotracheobronchitis)--showed a generally similar pattern of resolution irrespective of treatment. Nevertheless, seven out of 66 children receiving placebo were withdrawn from the trial with unremitting symptoms or complications thought to require antimicrobial treatment. Only two of 56 children receiving amoxycillin and none of 75 receiving co-trimoxazole were withdrawn. Three other children receiving amoxycillin and three receiving placebo were seen during the trial but further treatment was not thought to be necessary. Thus the return consultation rate in children receiving placebo therapy was 15% compared with 4% for those receiving antimicrobial treatment. Antimicrobial treatment was associated with less nasal discharge on the eighth day of treatment. Placebo treatment allowed an earlier return to normal activity. There was a high incidence of possible side effects on all regimens including placebo. It is concluded that the benefits of antimicrobial treatment in presumed viral respiratory infections are marginal, and they should not be routinely prescribed for these conditions.


On the occurrence of substance P-containing fibers in sympathetic ganglia: immunohistochemical evidence. Several sympathetic ganglia of the guinea pig, cat and rat were studied with indirect immunofluorescence technique using antibodies to substance P (SP) and dopamine-beta-hydroxylase (DBH), the latter of which represents a marker for noradrenaline containing neurons. In all ganglia studied SP-like immunoreactivity could be observed in certain nerve fibers (SP-positive fibers). Their number, localization fluorescence intensity and 'fluorescence morphology' varied considerably between the different ganglia as well as between species. In the inferior mesenteric ganglion and the coeliac-superior mesenteric ganglion complex of all species, and in particular of the guinea pig, a dense plexus of varicose SP-positive fibers was observed around the mostly DBH-positive, principal ganglion cells. In contrast no such fibers were seen in relation to the DBH-positive SIF cells of the ganglia. No SP-positive ganglion cells were observed in any of the ganglia studied. Previous biochemical and immunohistochemical studies have indicated that peripheral SP-positive fibers may represent the branches of primary sensory neurons. The rich supply of varicose, SP-positive nerve terminals, often surrounding the noradrenergic ganglion cells in a basket-like manner, may indicate that these fibers may play a functional role within the ganglia. Instead of merely passing through the ganglion they may be involved in special types of reflex arches.


Facilitation of the lordosis reflex in female rats by electrical stimulation of the lateral vestibular nucleus. (1) Electrical stimulation of the lateral vestibular nucleus (LVN) facilitated lordosis reflex responses to somatosensory stimulation, in estrogen-primed female rats. (2) Unilateral LVN stimulation was sufficient for this effect. Currents as low as 10muA could be used. (3) With the stimulus parameters used, the effect develops within 5 min of LVN stimulation and decays within 10 min following cessation of stimulation. (4) Together with previous evidence that LVN lesions disrupt lordosis, these results suggest LVN facilitation of spinal reflex mechanisms for lordosis.


Matrix vesicles in newly synthesizing bone observed after ultracryotomy and ultramicroincineration. Matrix vesicles were observed in femurs of 8-day-old chick embryos prepared by ultracryotomy. Some of the sections were subjected to ultramicroincineration. The unfixed tissues never came into contact with solutions, and thereby artifacts due to dissolution, redistribution, or rearrangement of the mineral constituents were avoided. In the osteoid, electron dense objects with the size and appearance of matrix vesicles were seen, although limiting membranes were not visible. After ultramicroincineration the vesicles were observed to contain small crystals and a less dense amorphous mineral material which may te the precursor of bone mineral. In addition, a ring of ash enclosed the crystalline and amorphous mineral and appeared to occupy the position of the vesicle membrane as seen in conventionally prepared material.


Effect of positive end-expiratory pressure on lung mechanics during anaesthesia in dogs. We studied supine anaesthetized dogs to determine whether the use of positive end-expiratory pressure to increase functional residual capacity (FRC) could thereby prevent the small lung compliance values frequently reported during anaesthesia. We first made control measurements of FRC and of both static (CSTAT) and dynamic (CDYN) lung compliance. Continuous positive pressure ventilation (CPPV) with 5 cm H2O positive end-expiratory pressure was then commenced, and FRC and compliance measurements were repeated at 15-minute intervals during one hour of CPPV. We found that the positive end-expiratory pressure served to increase FRC to a level expected for unanaesthetized normally breathing dogs in the same posture (J. Appl. Physiol. 36: 93, 1974), and this FRC was well maintained during the one-hour observation period. Despite the volume preservation, we found a marked decline in lung compliance, equal in both CSTAT and CDYN; however, these changes were completely reversed to control values following hyperinflation of lung (artificial sighting). We conclude that diminution in lung compliance in our anaesthetized dogs was not dependent on low lung volume breathing but must have been due to another mechanism such as suppression of sighing or possibly regional hypoventilation. It might well be important to recognize that reversible reduction of compliance may occur despite increase of FRC in patients managed with positive end-expiratory pressure.


Glucagon from avian pancreatic islets: radioreceptor studies. Glucagon extracted from isolated islets of the pigeon was studied by means of Sephadex gel filtration. Radioreceptor assay, using rat liver plasma membranes and radioiodinated porcine glucagon, showed that the bulk of the activity eluted with glucagon (molecular weight 3500). Avian glucagon appeared to be less effective than porcine glucagon in inhibiting the binding of labeled porcine glucagon to rat plasma membranes.


Immunofluorescence studies of Nitrobacter populations in soils. Certain steps of a protocol to enumerate a bacterium directly in soil by immunofluorescence were studied with respect to the enumeration of Nitrobacter in soils of diverse properties. Maximal counts of Nitrobacter were obtained by varying factors involved in the release of bacteria from the soil. Differences with respect to these factors were related to soil-colloidal properties. Enumeration protocols modified with regard to soil properties were used with strain-specific fluorescent antibodies (FA) to enumerate Nitrobacter populations (a) in soil during storage, (b) in comparison with most probable number (MPN) enumeration, and (c) occurring in a spectrum of soil samples. FA counting was rapid and precise and gave counts generally 10- to 100-fold higher than obtained by MPN. Nitrobacter cells of the two serotypes studied, designated as agilis and Winogradsky, occurred in each soil at levels of 10(4)-10(5) per gram.


A single-bottle blood culture system: evaluation and comparison with two other systems. A commercially available single-bottle blood culture system was evaluated at Ben Taub General Hospital, a Harris County District Hospital. Blood cultures from 1010 patients were examined with the Lederle Diagnostics one-bottle blood culture medium-SPS, Columbia broth (E-Vac, Pfizer), and an in-house-prepared brain heart infusion broth with p-aminobenzoic acid (PABA) and 0.1% agar. Of the 1010 patients examined, blood cultures from 211 (20.8%) were positive, yielding a total of 23 different species of microorganisms. Comparison of the results during clinical evaluation, as well as those from simulated blood cultures, showed that the Lederle Diagnostics blood culture bottle was as effective as the in-house-prepared brain heart infusion and commercially available Columbia broths for isolation of aerobes as well as anaerobes. The techniques used in the evaluation and the advantages of a single-bottle culture system are discussed.


Lipids of antibiotic-resistant and -susceptible members of the Enterobacteriaceae. Lipids of antibiotic-resistant and related -susceptible strains of the Enterobacteriaceae were extracted with chloroform-methanol and characterized by thin-layer chromatography, densitometry, and fatty acid analysis using gas chromatography. Quantitative differences which correlated with antibiotic resistance existed among the phospholipids and fatty acids. A relatively higher concentration of a ninhydrin-positive phospholipid concentration with a lower amount of phosphatidylethanolamine was observed in antibiotic-resistant strains of serratia marcescens. Bacterial strains which harbored R-factor 222 had a higher ratio of phosphatidylglycerol to diphosphatidylglycerol than their respective parent strains while those strains which were resistant to the polymyxins had a lower ratio of these phospholipids. Differences in the relative amounts of certain unsaturated and cyclopropane fatty acids were observed between susceptible and resistant strains. Such differences, however, were dependent upon a particular genus and species.


Reduced synthesis of beta-galactosidase in Escherichia coli infected with phage phi X 174. The synthesis of beta-galactosidase (EC 3.2.1.23;beta-D-galactoside galactohydrolase) in E. coli was repressed as a result of infection with single-stranded DNA phage phi chi 174. Evidence is presented to show that this repression was not due to the restricted entry of the inducer molecules into the infected cells but to some phage-specified product(s). It was further shown that either the infected cells synthesized a fewer number of enzyme-specific mRNA or all such molecules were translated with a reduced efficiency; the half-lives of the mRNA's remained more or less unaffected.


Structural proteins of a lipid-containing bacteriophage which replicates in Escherichia coli: phage PR4. PR4 is a lipid-containing bacteriophage which is able to replicate in Escherichia coli. The virus was labeled with either [14C]leucine and [14C]threonine or H235SO4 and then purified by several rounds of sucrose gradient centrifugation. Autoradiographs showed the virus to be composed of six major protein species with molecular weights (1) 68 000, (2) 47 500, (3) 38 500, (4) 35 000, (5) 20 700, (6) 16 500 daltons. Electropherograms showed protein No. 2 to be the major protein, comprising about 43% of the total weight of viral protein.


Isolation and distribution of oligotrophic marine bacteria. A useful plate culture method for isolating oligotrophic bacteria found in the low-nutrient environment of the open sea has been developed. The method uses a glass-fiber filter substitute for agar. Nutritional requirements of oligotrophic bacteria consisted of a dilute mutrient solution containing 16.8 mg C/l total organic carbon aseptically added to the sterilized filter. Distribution of bacteria in oceanic and neritic seawater was determined using the membrane filter method. In the case of seawater containing less than 0.5 mg/l dissolved carbohydrates, plate counts of oligotrophic bacteria were found to be several- to 100-fold greater than the heterotrophic bacterial counts enumerated by standard methods routinely used for enumeration. However, in seawater containing approximately over 0.5 mg/l dissolved carbohydrates, heterotrophic bacterial counts were 10-fold greater than oligotrophic bacterial counts.


Transient monocular blindness.

Transient monocular blindness. This paper is a review of the causes of intermittent monocular blindness. The nature of cholesterol and platelet retinal emboli is discussed. Their sources, the frequency with which they may cause transient or fixed blindness and the association between these emboli and pathology of the major cerebral vessels and other organs is discussed. Consideration is given to the equally important abnormalities of platelet behavior and to some of the physiology of retinal blood flow and non-embolic blindness. The current treatment of this symptom may be anticoagulation, surgical correction of a stenotic artery or both. The effect of treatment is unpredictable and in some situations the rationale is suspect. This review may provide a summary on which to base future studies of the effectiveness of various therapeutic agents.


Alkaline gastritis and alkaline esophagitis: a review. Alkaline gastritis and alkaline esophagitis are now precisely defined syndromes. They occur most often after gastric surgery in which function of the pyloric and lower esophageal sphincter is compromised. Reflux of bile in these patients can then lead to severe inflammation of the gastric and lower esophageal mucosa. Epigastric pain, nausea and bilious vomiting are characteristic symptoms. Gastroscopy with biopsy is, therefore, the definitive diagnostic test; during endoscopy bile is seen in the lower esophagus or stomach, and the mucosa is red, friable and contains acute erosions. Conservative therapy including the administration of cholestyramine has not been helpful. Surgery consisting of diversion of the duodenal contents away from the stomach and lower esophagus is the treatment of choice. The Roux-en-Y procedure has been used most often and has resulted in the amelioration of the symptoms and signs in most patients.


Ocular complications in renal transplant recipients. The occurrence of ocular complications among 62 recipients of functioning renal transplants was reviewed 6 months to 7 1/2 years after transplantation. Posterior subcapsular cataracts were found in 29 (46.7%). A positive correlation was found between the development of cataracts and the total dose of corticosteroid, the number of days on which the dose exceeded 100 mg, and the number of rejection episodes treated with high doses of corticosteroid. Identification of specific high-risk periods in regard to cataract formation could lead to use of preventive therapy, as yet undetermined, during these periods. Lower doses of corticosteroids should be used to treat rejection episodes.


A comparison of cyclophosphamide, adriamycin, 5-fluorouracil (CAF) and cyclophosphamide, methotrexate, 5-fluorouracil, vincristine, prednisone (CMFVP) in patients with metastatic breast cancer: a Southeastern Cancer Study Group project. In an ongoing prospective randomized study, 113 evaluable patients have received either a three-drug combination that included cyclophosphamide, Adriamycin and 5-fluorouracil (CAF) or a five-drug combination including cyclophosphamide, methotrexate, 5-fluorouracil, vincristine and prednisone (CMFVP) given intermittently 1 week out of 4. Responses (64%), median duration of response (32 weeks), and median duration of disease control (32 weeks) achieved with CAF were superior to those achieved with CMFVP (37%, 22 weeks, 17 weeks, respectively). Morbidity secondary to CAF was significant, with nausea and vomiting, malaise, total alopecia, and granulocytopenia being the main features.


Mutagenicity of smoke condensates from cigarettes, cigars and pipe tobacco. Smoke condensates from cigarettes, cigars and pipe tobacco were mutagenic on Salmonella typhimurium TA100 and TA98 when activated with rat liver microsomal system. Mutagenicity of a unit weight of smoke condensate was rather high in cigars, low in pipe tobacco and intermediate in cigarettes. Specific mutagenic activity was almost comparable among smoke condensates from low- to high-tar cigarettes, although some variations were observed depending upon the country producing the cigarettes. Marked mutagenicity of cigarette smoke condensate could not be explained by the benzo (a) pyrene or nitroso compounds it actually contained, suggesting the presence of other very potent mutagens in tobacco smoke condensates.


Phase II study of oral methyl-CCNU and prednisone in previously treated alkylating agent-resistant multiple myeloma. Thirteen patients with multiple myeloma (MM) who either failed to respond to or who were relapsing from standard agents and who received four or more courses of methyl-CCNU + prednisone (adequate drug trial) are reported in this paper. Methyl-CCNU was given orally before breakfast at 6-week intervals at a starting dose of 50 mg/m2 with the intention of increasing the dose to 100, 150, and 200 mg/m2 with each subsequent course. The dose of prednisone was 75 mg/day x 7 with each course. The response rate was 46% (six of 13 patients). No patient had better than a fair response. Drug toxicity, severe enough to prevent further dose escalation, was observed in every case. Prior BCNU therapy or the lack of response to previous alkylating agents did not prevent a response to methyl-CCNU + prednisone. The response rate of methyl-CCNU + prednisone in MM is comparable to the results achieved with other agents in similar groups of patients.


Evaluation of a MOPP-type regimen in histiocytosis X--a Southwest Oncology Group study. The overall 38% response rate in histiocytosis X for cyclophosphamide, vincristine (Oncovin), prednisone, and procarbazine was not superior to responses that can be achieved with the component single agents. This is in contrast to the higher response rates that can be achieved by the combination of these agents in Hodgkin's disease. This is the second study of combination chemotherapy with individually effective agents that has failed to improve the response rates in histiocytosis patients with poor prognostic indicators.