Iatrogenic exfoliation of teeth by the incorrect use of orthodontic elastic bands.

Four cases of improper use of elastic bands for minor orthodontic treatment are described. For all the patients, the treated teeth were damaged and local surgical intervention was necessary. To prevent complications when elastic bands are being used, the dentist must provide stabilization of the elastic bands by brackets, bands, or bonded attachments, inform the patient about the possibility of losing the rubber elastic band, and mark the elastic rubber by radiopaque material. Every effort must be made to preserve the damaged anterior teeth by fixation and surgical treatment.


Isolation of mycobacteria from dairy creamery effluent sludge.

Sixty three samples of dairy creamery effluent were examined for the presence of mycobacteria. Thirty two strains were isolated from 27 samples. These were classified as follows: M. fortuitum (13), M. peregrinum (6), M. gordonae(5), M. marianum (scrofulaceum) (4), unidentified (4). Ten strains, representative of the groups isolated, were tested for their effect on experimental animals. None were pathogenic for guinea pigs or mice but a number produced a minimal amount of skin sensitization in guinea pigs to avian and mammalian tuberculin.


Prevention of low temperature denaturation injury in T4Bo phage by low concentrations of traditional cryoprotective additives.

The well known cryoprotective additives glucose, sucrose, glycerol, dimethyl sulphoxide, polyvinylpyrrolidone, dextran and ammonium acetate have been found to prevent inactivation of T4Bo phage frozen in sodium bromide solutions. Their protective effects in this experimental system could not be accounted for by a colligative mechanism. It is proposed that they may act by modifying the structure of the unfrozen aqueous phase rather than by direct interaction with the phage.


A clinical trial of WRL 105 strain live attenuated influenza vaccine comparing four methods of intranasal vaccination.

A single intranasal dose of 10(7-0) EID50 recombinant WRL 105 strain live attenuated influenza vaccine was administered intranasally to 193 volunteers either as nose drops or by one of three spray devices which produced sprays of differing physical characteristics. In volunteers with homologous haemagglutinating inhibiting antibody titres of less than or equal to 20 before vaccination, seroconversion rates varied widely from 80% following the administration of drops to 71%, 57% and 28% with the three spray devices. In the week following vaccination 16 (22%) of 74 volunteers who were found to show a fourfold or greater antibody response to took analgesics to control symptoms in comparison with 4 (7%) of 58 volunteers who exhibited no serological response to vaccination (P less than 0-05). However, neither the occurrence of upper respiratory nor systemic symptoms were significantly different in these two groups and the degree of attenuation of the recombinant WRL 105 strain appears to be acceptable for future use.


Platelet-induced tolerance in the production of heteroantisera to human leukemia-associated antigens.

A method is described for raising antisera to human leukemia cells of an individual patient in mice rendered tolerant with cyclophosphamide to platelets obtained from the same patient. The resulting antisera are able to distinguish serologically between leukemic blast cells and remission cells of patients with acute leukemia and may be recognizing leukemia-associated antigens. The antisera are similar in activity to antisera raised following  tolerance-induction with remission leukocytes, but larger volumes of anti-leukemia antiserum can be raised using the more easily obtainable platelets. This technique provides further evidence that human platelets share many of the antigens present on human leukocytes.


Acute infections in interferon-treated patients with osteosarcoma: preliminary report of a comparative study.

The incidence of acute infections in eight patients with osteosarcoma who are receiving interferon regularly is currently being compared with the incidence among their family members. Observations thus far indicate that the patients treated with interferon are less frequently and less severely ill than their untreated family contacts. The study is continuing, and the patients will also be observed after discontinuation of interferon treatment. Sera are regularly frozen for future viral serologic tests. The preliminary clinical data indicate that therapy with exogenous interferon may afford protection against common acute infections.


A solid-phase radioimmunoassay for the measurement of lactoferrin in human plasma: variations with age, sex, and disease.

A solid-phase radioimmunoassay is described for measuring lactoferrin levels in normal human plasma. The sensitivity of the assay was 6 ng. per milliliter with an intraassay coefficient of variation of 4 per cent and an interassay value of 9 per cent. Healthy adult males had a mean plasma level of 1.62 mug per milliliter which was significantly higher than adult females, 1.07 mug per milliliter. Postmenopausal females had levels similar to men, 1.74 mug per milliliter, while younger women had a significantly lower mean value, 0.75 mug per milliliter. Two menstruating women and 2 pregnant women had moderately elevated levels. Consistently elevated levels were found in patients with untreated or relapsing chronic myeloid leukemia--all over 12.0 mug per milliliter, while patients on marrow suppressant therapy tended to have subnormal levels. The collection of serum specimens as opposed to plasma, resulted in inconsistently elevated levels: EDTA was the anticoagulant of choice, as heparin interfered in the radioimmunoassay system.


Sex dependence of the metabolic activation in vitro of the mycohepatocarcinogen aflatoxin B1.

Using liver microsomes, metabolic activation of aflatoxin B1, determined by the formation of DNA- and protein-binding metabolites, was studied in an effort to elucidate the possible basis underlying sex differences in sensitivity of the male and female rat to aflatoxin B1. Male rats, which are more sensitive to AFB1, produced significantly higher amounts of these metabolites than female rats; castration of the male reduced the level to that of the female, whereas ovariectomy was without any effect.


Acute pulmonary edema and hyperosmolality: a clinical study.

Patients in the coronary care unit with acute pulmonary edema, heart failure, and other organic heart disease were studied. Blood and urine samples were taken on admission prior to any treatment and later at prescribed intervals. All the patients with APE were found to have elevated plasma osmolalities and hyperglycemia on admission which decreased with treatment. This was in contrast to the other two groups excluding those factors such as ethyl alcohol and diabetes which can raise plasma osmolality or blood glucose. A discussion of this mild hyperosmolal state in APE follows including possible causes as well as cellular effects of hyperosmolality on humans.


Alkali-labile regions and strand breaks in DNA from cells treated with daunorubicin.

Lymphocyte cells (P-288) in culture were exposed to daunorubicin (Dm) and analyzed for DNA damage by means of alkaline sucrose and neutral (formamide) sucrose gradients. In neutral gradients, the sedimentation rate of DNA from cells treated with 25 X 10(-8) M Dm for brief periods up to 60 min is the same as that of the DNA from control cells (i.e., drug-treated for "zero" time). In contrast, extensive DNA damage is already apparent in alkaline gradients after treatment of the cells for only 30 min. After treatment for 2 hr, single-strand damage is evident in both types of gradient. These results demonstrate the introduction of alkali-labile regions in the DNA of cells treated for 45 or 60 min with Dm. The DNA damage is concurrent with inhibition of thymidine incorporation, and both may occur at threshold levels without affecting subsequent growth of the P-288 cells after short exposures to low concentrations of the drug.


Uptake of adriamycin and daunomycin in L1210 and human leukemia cells: a comparative study.

The uptake of Adriamycin (AM) and Daunomycin (DM) in L1210 leukemia cells was found to be dependent on drug dose, time, and temperature. DM was taken up to a greater extent in human leukemia cells in vitro and in L1210 leukemic cells both in vitro and in vivo. Intracellular retention of DM was less than that of AM. Drug transport appears to be a factor in developing resistance to DM in L1210 cells, but the data for AM were inconclusive in this regard. The importance of cellular uptake and retension operative in determining differences in the overall therapeutic efficacy in vivo remains to be elucidated.


Response of murine DNA polymerases C and N to partial hepatectomy.

Two DNA polymerases, designated C and N have been characterized in extracts of murine spleen and liver (Evans et al. 1974). The effect of partial hepatectomy on the activity levels of these two enzymes was studied by gel filtration of crude extracts on Sephadex G-200. Both enzymes were observed to respond to the proliferation stimulus. Polymerase N activity reached maximum levels between 30 and 48 hours after hepatectomy. Polymerase C response was delayed until 30 hours and reached maximum values at 48-72 hours.


The effect of pretreatment with allogeneic fetal cells on hamster melanoma.

Pretreatment with 7-day embryonic tissue mixed with incomplete adjuvant retarded the growth of melanoma transplants in the Syrian hamster. This effect was not apparent in animals treated with fetal tissue suspension, only or with fetal tissue and complete adjuvant. The inhibitory effect on tumor growth noted in animals challenged with tumor cell doses of 5 x 10(5) or 2.5 x 10(6) cells was abolished when the tumor dose was increased to 5 x 10(6). These results suggest that hamster melanoma and fetal tissue may share a common antigen.


Dose distribution of therapeutic electron beams and automation of treatment planning.

Until the present time, there has been no mathematical evaluation of the therapeutic electron beams in tissue material. Electron treatments are becoming universally applied from betatrons and linear accelerators. There is, therefore, a need for dosimetric programming of hospital computers for electron treatment planning such as those used in photon beam dosimetry. In this study, we have developed a theoretical model for dose calculation in a clinical therapeutic electron beam (Osman, 1972 and 1973). On the basis of this proposed theory, one can predict dose profiles at any source to skin distance "SSD"' and at any depth in tissue "X". Our theoretical model is based on considering the clinical broad electron beam used in radiation therapy as being made up of an infinite number of identical and initially parallel pencil beam components, each of minute width, to which the existing theories (Lewis, 1950; Attix et al., 1968) on electron multiple scattering, in the concerned medium apply. Dose profiles in tissue, as obtained from this model, could provide useful information as input data for routine programs of treatment planning with high energy electron beams, using mini-computers. Further, it is also possible to account for any body inhomogeneity such as subcutaneous fat, lung, air volumes, body cavities, fluids and bone from the basic parameters of these media.


Effects of radiation and porphyrin on mitosis and chromosomes in human hematopoietic cell lines.

The effect on mitosis of a human hematopoietic cell line RPMI-1788 treated with a metal chelate (Zn++) of meso-tetra (p-carboxyphenyl) porphine (Zn-TCPP) alone at various concentrations or in combination with gamma-irradiation at various doses were studied. The results showed that both Zn-TCPP and radiation were effective in interfering with normal mitosis and that the effect of radiation was relatively more effective. Data also suggest interacting effects between Zn-TCPP and gamma-irradiation. At low doses of radiation, Zn-TCPP potentiated the effect of radiation. The reverse seemed to be true at a high dose of radiation. The effects of two porphyrins (Zn-TCPP and hematoporphyrin) and radiation on chromosomes were also studied. Chromosomal aberrations characteristic of radiation were observed. The porphyrins were found not to be effective chromosome-breaking agents under the experimental conditions tested.


Tensile strength, relaxation and mechanical recovery in rat skin as influenced by maturation and age.

Several mechanical parameters were measured in skin strips of rats at various ages (1, 2, 4, 12 or 24 months). Ultimate load and tensile strength increased during the maturation process and reached a maximum at 12 months. In old animals (24 months) a decrease of these parameters was noted. After various preloads relaxation was measured. Relaxation could be described mathematically as a function of logarithm of time. The relaxation coefficient decreased during the maturation as well as during the ageing period. After unloading up to 1/5 initial load mechanical recovery was measured. Mechanical recovery was increased depending on age during the whole life span. It is concluded that various components of viscoelatic behaviour are influenced differently by the maturation and the ageing process.


Correlation of response to 1-beta-D-arabinofuranosyl cytosine and metabolism of drug by tumor.

Phosphorylation versus deamination of ara-C by tumor homogenates was measured in 10 patients prior to treatment with ara-C infusion. The ratio ranged from 0.24 to 1.2 in 5 malignant melanomas, 2.6 to 3.2 in 3 histiocytic lymphomas and 0.9 in a hemangiopericytoma. Treatment of the 9 patients with 2 courses of 5-day continuous ara-C infusion failed to produce objective evidence of tumor regression. The tenth patient had lymphosarcoma leukemia. The baseline ratio of ara-C phosphorylation over deamination activity of the tumor cells was 1.23. Treatment with 2 courses of continuous ara-C infusions produced a brief state of complete remission. The ratio of araC phosphorylation over deamination activity in the tumor cells after relapse was only 0.1 and retreatment with ara-C infusions failed to produce antitumor response. This study indicates that factors other than a high ratio of phosphorylation over deamination activity by the tumor seem to play an important role in the susceptibility of tumors to ara-C treatment.


The Robin anomalad - its nonspecificity and associated syndromes.

The Robin anomalad, once thought to constitute a specific syndrome, is now known to occur with several genetic syndromes, a few drug-induced syndromes, and some loosely associated anomalies. The mode of inheritance in genetic syndromes with the Robin anomalad is that of the particular syndrome with which the anomalad is associated.


Oral changes in Crohn's disease.

Oral lesions in patients with Crohn's disease may be characteristic granulomatous lesions, secondary results of deficiency conditions, or coincidental lesions. Thorough investigations are necessary to rule out involvement of the gut in the disease process. If results fail to substantiate a diagnosis of Crohn's disease, regular follow-up examinations are necessary to detect possible later development of the disorder.


Inappropriate secretion of antidiuretic hormone after cerebral injury.

A case has been presented in which a patient sustained a closed head injury with concomitant maxillofacial injuries; early signs of water intoxication and ISADH developed six days after injury. This disorder was corrected by restricting free water intake for six days until equilibration occurred. Successful reduction of the facial fractures was accomplished after stabilization of the patient's neurological condition and correction of her metabolic disorder. The ISADH and resulting hyponatremia have been documented in a variety of disease states including trauma to the central nervous system. Disruption or irritation to the hypothalamic-neurohypophyseal system has been proposed as the mechanism of dysfunction after cerebral injury. The results of the secretion of inappropriate amounts of ADH relative to renal function and homeostatis have been discussed. Clinical and laboratory diagnosis as well as the elective and emergency management of ISADH have been reviewed. The fact that the sequelae of this abnormal metabolic state may mimic or mask the neurological deterioration which may follow cerebral injury is significant. This may contribute to the difficulty in making a correct diagnosis and designing proper therapy. The problem is basically one of differentiating a correctable metabolic disorder from a lesion that can be fatal unless surgically removed.


Treatment of central hemangioma of the maxilla by embolization: report of case.

Some methods of treatment for cavernous hemangiomas are reviewed and briefly discussed. Artificial embolization with barium-impregnated spheres was used by us because any other method of treatment seemed to be inadvisable. Techniques of the surgical procedure were followed as programmed by Lussenhop and others, 12 Longacre and others, 2 and by Hoey and others.9 A 47-month follow-up with no radiographic evidence of residual hemangioma has indicated a satisfactory result in this case.


Unilateral relapse after oblique osteotomy of the mandibular rami: report of case.

A case of unilateral relapse of mandibular prognathism after oblique osteotomy of the mandibular rami is described. A brief review of the pertinent literature is presented and a method of intraosseous fixation by Vitallium plating is illustrated. The need for close postoperative observations and reinstitution of active therapy at the first sign of a relapse tendency is emphasized.


Correction of class II malocclusion by combined sagittal ramus and subapical body osteotomy.

The complete mandibular subapical osteotomy has been presented as a method for correction of Class II malocclusions. At this point, it holds some promise as a solution to the skeletal relapse seen in the surgical correction of Class II malocclusions with use of other techniques. The procedure is a straight-forward combination of the sagittal osteotomy as described by Obwegeser and the subapical osteotomy as described by Köle. Further investigation into the versatility of this procedure is recommended.


Epidural hematoma after minor oral trauma.

A case report was presented in which a 15-year-old boy was beaten about the head with a baseball bat. Intraoral trauma and facial lacerations were repaired. Since results of the neurological examination were within normal limits, the patient was discharged. The next day, the patient became lethargic; however, the patient's mother did not bring the patient back to the hospital until the routine postoperative visit. At that time, the patient had right hemiparesis, was unable to speak, and was clearly obtunded. A carotid angiogram disclosed a left venous epidural hematoma in the parietal area. A craniotomy was performed with good results. The importance of follow-up neurologic examinations in cases of trauma to the face and head is stressed.


False aneurysm of the facial artery.

A false aneurysm with typical history, clinical and radiographic findings, and histological picture is reported. The absence of enlargement since onset and the absence of bruit were atypical. The latter was probably due to the thick wall of the aneurysm. False aneurysm should be part of the differential diagnosis of swelling of the jaws. Aneurysms should be treated surgically because of the potential for progressive enlargement with danger of spontaneous rupture.


Osteosarcoma of the thyroid gland.

A case of osteosarcoma of the thyroid gland in a 61-yr-old woman who died from tracheal compression due to local recurrence 6 wk after operation is presented. The main features of 27 previously reported similar cases, and the problems of histogenesis are discussed.


Electrical stimulation of osteogenesis in pathological osseous defects.

Although the literature on bioelectric phenomena is extensive, there is a paucity of research on its application in the field of dentistry. This study was designed to evaluate the effect of bioelectrical stimulation on pathological osseous defects in the periodontium of Beagle dogs. Galvanic and thermocouple generators were designed to deliver a current in the nanoampere range. The galvanic unit produced a constant direct current. The thermocouple unit was designed to deliver an oscillating dc current. The generators were implanted in the area of periodontal osseous defects in five dogs. These sites were randomly designated as experimental, active controls, and passive controls. The experimental and control units were implanted submucoperiosteally with the negative electrodes extending into osseous defects. The bone was labeled with Procion Red H-8BS at 1 week. Clinical and radiographic evaluation revealed no significant changes in the depth of the defects. The animals were sacrificed at 45 days and block sections were prepared for histological measurement. From the results of this study it was concluded: 1. Induced constant or oscillating negative dc current levels in the nanoampere range were unsucessful in causing regeneration of periodontal osseous defects either clinically or radiographically in the Beagle dog over a 45-day period of application. 2. Constant or oscillating negative dc current levels in the nanoampere range did not stimulate a significant amount of periosteal osseous apposition. 3. Oscillating negative nanoampere dc currents produced an increase in endosteal osseous apposition as compared to controls. 4. Constant negative dc current in the nanoampere range produced an increase in endosteal osseous apposition as compared to controls. 5. The histological density of the bone in the area of pathological periodontal defects was increased in all experimental sites as compared to controls. 6. Devices of comparable size and construction to both the galvanic and thermocouple electronic units used in this study are acceptable for future investigation of the effect of bioelectrical stimulation on osteogenesis in alveolar bone.


Vital root amputation. A clinical and histological study.

An investigation was designed to study the clinical and histological effects of delaying endodontic therapy for a period of 2 weeks without dressing of the exposed pulp, after vital intentional root amputation had been carried out on periodontally involved teeth. From the results obtained it was possible to conclude that: 1. Endodontics, prior to surgical root amputation, remains the treatment of choice. 2. When preoperative endodontics is not possible, the approach used in this study will provide good results without severe, adverse clinical or histological effects. 3. The altered periodontal condition and tooth form do not interfere with the performance of definitive endodontics 2 weeks after surgery. 4. Definitive preparation of the cut tooth surface at the time of root amputation facilitates maintenance of oral hygiene after removal of the periodontal dressings. 5. The high percentage of involvement of the distobuccal root of the first maxillary molar in periodontal situations requiring correction by root amputation, may be due to the anatomy of this root.


Study of gliding tooth contacts during mastication.

Tooth gliding occurs frequently both in the opening (55.9%) and closing (60.5%) chewing strokes. The average glide length was 1 mm. Good occlusions could not be distinguished from poor occlusion on the basis of frequency and length of gliding contacts. The greater the component of lateral movement during the closing stroke, the greater the probability that gliding contacts would occur.


The destructive potential of electrosurgery on the periodontium.

Three cases have been presented illustrating the complications which may arise from the use of electrosurgery. These cases provide clinical evidence of the potential for severe periodontal destruction, a potential that must be weighed carefully by any practitioner who uses an electrosurgical instrument. It is suggested that new scientific studies be initiated to establish basic principles that will eliminate the hazards that now exist in the routine use of electrosurgery.


Vitamin E and periodontal disease.

Vitamin E levels in patients with and without periodontal disease were measured. There was no significant difference in the levels of serum vitamin E between these two groups. These data do not provide support for the treatment of inflammatory periodontal disease with vitamin E.


Metastatic adenocarcinoma of gingiva. Report of a case.

A case of metastatic neoplasm developing in the gingiva from a primary lesion, probably sited in the gastric mucosa, has been reported. In this case, the first demonstrable metastatic lesion was located in the gingiva. The dentist has a great responsibility in detecting malignancy in the oral cavity because of his opportunities during routine examinations.


The effect of urea peroxide on dilantin hyperplasia.

1. A significant inverse relationship was found between the severity of the hyperplasia and the duration of SDH. 2. The severity of hyperplasia and in flammation was found to vary significantly and inversely with oral hygiene scores. 3. Tooth brushing twice a day and the application of Oxygel showed no significant clinical changes on the inflammation and hyperplasia in the test group, whereas the control group using a placebo showed a moderate reduction in gingival inflammation.


Tolerance and limited cross-tolerance to the effects of N, N-dimethyltryptamine (DMT) and lysergic acid diethylamide-25 (LSD) on food-rewarded bar pressing in the rat.

Lysergic acid diethylamide=25 (LSD) and N,N-dimethyltryptamine (DMT) abolish food-rewarded, fixed-ratio bar pressing by rats in a dose-related fashion. Adult male Holtzman rats trained to press a bar (respond) for milk reward on a 4-response fixed-ratio schedule were given i.p. injections of 3.2 or 10 mg/kg of DMT every 2 hours for 21 days. Every 24 hours the animals were placed in operant chambers for 30 minutes before a scheduled injection and were left in the chambers for 30 to 80 minutes after. During the first week of chronic treatment, daily bar pressing worsened progressively until the 6th day of the series, at which time rats in the 10 mg/kg group did not bar press at all. As the chronic injections continued, rates of bar pressing gradually increased until responding was not disrupted at all by an injection of DMT. Rats in the 3.2 mg/kg group showed cross-tolerance to an injection of LSD (0.1 mg/kg). Another group of rats was made partially tolerant to the disruptive effects of LSD (0.1 mg/kg i.p.) on bar pressing with a series of injections given once per day for 21 days and then three times per day for the next 4 days. Cross tolerance was not demonstrated to a challenge injection of 10 mg/kg of DMT. The LSD injections were continued for another 3 to 5 days until the animals were completely tolerant to the LSD. They then displayed cross-tolerance to 3.2 mg/kg of DMT.


[A respiratory pump for small animals (author's transl)].

A respiratory pump is described which is controlled by compressed air and contains no mechanical moving parts, neither a motor nor a piston. It can function on a self-adjusting rhythm or can be regulated externally (for example by the discharge of a phrenic nerve).


Amniotic cell 4-methylumbelliferyl-alpha-glucosidase activity for prenatal diagnosis of Pompe's disease.

Using a simple fluorometric assay for alpha-glucosidase activity of cultured amniotic cells, we have monitored two pregnancies from families at risk for Pompe's disease. The fetus was judged to be affected in one, the pregnancy being terminated and unaffected in the other. The accuracy of these predictions was confirmed. These results suggest that this assay allows accurate prenatal diagnosis of Pompe's disease, three weeks after diagnostic amniocentesis.


Proliferation and production of hemopoietic cells two stages of disease: preleukemia and overt leukemia.

The kinetics of erythropietic and granulocytopoietic cell proliferation have been investigated in the same patient at two distinct stages: firstly in preleukemia presenting as pancytopenia with ineffective erythropoiesis, and secondly 2 years later in acute myelogenous leukemia. The method investigation is based on determining the DNA synthesis rate of individual cells by means of quantitative 14C-autoradiography after short-term incubation with 14C-thymidine and fluorodeoxyuridine. Erythropoiesis was equally ineffective in the two stages, the rate of proliferation, however, slowed down towards the leukemic state. The production rate of myeloblasts was markedly reduced in preleukemia accompained by a very low labelling index. In leukemia on the other hand the production rate was increased to such a degree that the leukemic myeoblast compartment is to be considered as prevailingly self-reproductive. The proliferation rate of myeloblasts was reduced already in preleukemia, and there was a further decrease in leukemia. From the point of view of cell kinetics the manifestation of leukemia in this patient is explained best by a change in the mode of proliferation: the myeloblasts change from steady state growth to behaving like an exponentially expanding population.


In vitro granulocytic stem cell clonogenic capacity of marrow and spleen cells from mice infected with Rauscher leukemia virus.

Bone marrow and spleen cells from early, midstage, and terminal Rauscher leukemia virus (RLV-A)-infected erythroleukemic mice were assessed for granulocyte stem cell (CFU-c) clongenic capacity in the semisolid agar culture assay. It was found that marrow CFU-c concentrations exceeded normal in early stages of this erythroid disease but returned to near normal values during mid- and terminal phases. Splenic CFU-c concentrations, on the other hand, were generally higher than control values for all stages of the disease. These results are discussed with reference to the pathogenesis of RLV-A disease.


Platelet granulopathy: a new morphologic feature in preleukemia and myelomonocytic leukemia: light microscopy and ultrastructural morphology and cytochemistry.

We have previously reported on the ultrastructure of platelets in preleukemia and myelomonocytic leukemia. We referred to an unusual and distinct anomaly of the platelet granules found in 15 of 16 patients. In the present communication we wish to describe and illustrate the light microscopic appearance of giant anomalous granules. Close scrutiny of the platelet morphology and a search for the aforementioned platelet granulopathy are important in the evaluation of patients with myeloproliferative diseases. In this paper we describe and illustrate in more detail the ultrastructure and ultrastructural histochemistry of the abnormal granules. In those patients with the platelet granulopathy, we have conducted in vitro platelet aggregation studies and carried out an electron microscopic evaluation of the aggregates. At least some of the giant granules remained morphologically intact in advanced stages of the aggregation phenomenon, and thus they are probably composed of elements that were not released during aggregation.


Cell kill kinetics with hydroxyurea.

The effects of various concentrations of hydroxyurea (HU) on a human lymphoid cell line in exponential growth phase have been studied using a combination of methods, including determination of the total and viable cell counts; the cells relative DNA content, measured in a flow microfluorimeter after staining with a fluorescent Feulgen technique; the mitotic index; and the percentage of cells incorporating thymidine-3H (TdR-3H) during brief and continuous exposure to the isotope both in the presence and absence of colcemid. A significant redistribution of the cells in the various phases of the cell cycle occurred during the first 24 hr of continuous treatment with 10(-3) M and 10(-2) M HU as follows: (1) division of cells in G2; (2) depletion of mid and late S phase cells due to early cell death; (3) movement of most G1 cells at a normal rate into early S phase where they accumulate; and (4) arrest of the remaining cells in G1, which represented the surviving population after treatment for 96 hr or longer. After removal of the drug, the cell fraction blocked in early S phase progressed semisynchronously through S, but many of the cells were unable to complete division. Their capacity to recover depended on the drug concentration and duration of exposure, but in general the cellular injury caused by HU was more reversible than that caused by "equivalent" concentrations of arabinosylcytosine.


Usefulness of cytosine arabinoside (NSC-63878) and prednisone (NSC-10023) in refractory childhood lymphoblastic leukemia.

One hundred forty-three children with refractory lymphoblastic and undifferentiated leukemia (ALL/AUL) were treated with cytosine arabinoside (Ara-C) and prednisone (Pred). The dose and duration of Ara-C was escalated during induction depending on the response seen in the peripheral blood and/or bone marrow. For those achieving a remission, Ara-C was also used to determine its maintenance capabilities. Of the 143 children, 79 attained a clinical remission, 45 having a complete bone marrow remission and 34 having a partial remission. Maintenance of remission with twice weekly Ara-C was short and did not appear to depend on the amount of Ara-C given during induction. The major toxicity of Ara-C was myelosuppression.


Clinically significant cardiac infiltration in acute leukemia, lymphocytic lymphoma, and plasma cell myeloma.

Cardiac infiltration by hematologic neoplasms leading to clinically significant cardiovascular disease is rare. Three such cases are described in this report, and it is suggested that rare manifestations of hematologic neoplasms may become more common in the future since these diseases are more amenable to therapy than heretofore. Cardiac involvement with hematologic neoplasms is of more than academic interest since this complication is likely to respond to radiotherapy.


[Association of leukemias and tumors. Studies of 502 cases of leukemia].

Nineteen association of leukaemia and tumour were noted in a series of 502 cases of leukaemia: 12/180 (6.6%, compared with 4.7% of 5136 cases in the liteature) for Chr. L.L. (hypogammaglobulinaemia, reduction in single Ig. serious herpes zoster and the T-lymphocyte nature of leukaemia were not more frequent in these associations); 2/102 (1.9%, compared with 2.6% of 1267 cases in the literure) for Chr. M.L.; 5/220 (2.2%, compared with 2.19% of 1138 cases in the literature) for A.L. The mean age of the overall leukaemia series was virtually the same for A.L. (47 yr in a group composed of subjects aged over 12 yr) and Chr. M.L. (48 yr), with the same incidence of association (2.2 and 1.9%), whereas it was 64 yr and 6.6% incidence in Chr. L.L. The bilogarithmic increase in the incidence of tumours with age may itself explain the higher incidence of Chr. L.L. associations. The duration of leukaemia and the age of incidence must be taken into account in any discussion of the significance of such associations.


Noise as a health hazard in the boilermaking industry.

An investigation was made into noise hazard aspects of three boilermaking firms. This consisted of noise measurement of both background and impact noises, of field audiometry of the workforce and a questionnaire. Environmental findings so far show the noise to be hazardous to hearing. Audiometric testing revealed a high rate of hearing impairment. A general lack of knowledge of noise hazard was shown, and the need for a hearing conservation programme demonstrated.


Spironolactone-associated aggravation of renal functional impairment.

Five patients in whom deterioration in renal function was associated with the use of the aldosterone antagonist, spironolactone, are reported. Care in the use of this drug is urged, especially in the elderly. Patients with known renal functional impairment should not receive this agent.


Management of sugar intolerance in children.

Sugar intolerance is a common problem in paediatric practice. The usual type is lactose intolerance following gastroenteritis, but it may also occur in a wide variety of disorders of the small bowel. Diagnosis depends upon identification of reducing substances in the stools. An approach to dietary management of lactose intolerance is given and use of a carbohydrate-free formula in secondary monosaccharide intolerance is described. In each situation, threshold for digestion or absorbtion of carbohydrate is approached gradually from below, and overflow detected by Clinitest stool testing.


A short interview as an assessment of student health and potential.

A prospective study of the effectiveness of a short interview in assessing health needs and potential performance of students. The results indicate that such an interview is of limited value in the recognition of the vulnerable student, can predict performance of the best students with reasonable accuracy, can recognise potential drop-outs, but failed to distinguish in a borderline group between those likely to succeed and those not.


Vibrio parahaemolyticus-food poisoning: case report.

Symptoms of food poisoning occurred following the ingestion of raw shellfish purchased in the Auckland area. Vibrio parahaemolyticus was recoverer from the patient. The potential of this and closely related microorganisms to cause illness is reviewed.


Pull-through uretero-lithotomy.

A method for removing a stone from the lower end of the ureter by a pull-through uretero-lithotomy is described.


Self poisoning in Auckland.

The records of all deliberate self poisoners seen at Auckland Hospital in a 15 month period in 1971-72 were examined. There were 550 patients of whom 35 presented twice and six three times. Five persons died. A subgroup of 100 was interviewed personally and compared with 100 orthopaedic outpatients matched for age and sex. Self poisoners are more likely to be aged 15-24, of female sex, have seen a psychiatrist, self poisoned previously, come from disturbed family and marital backgrounds and show other signs of moderate maladjustment. Drugs taken are usually prescription drugs obtained legitimately with psychiatric ones particularly hypnotics and anxiolytics, predominating. The act is usually impulsive and not designed to result in death. Most patients require only outpatient treatment. It is suggested that the term attempted suicide is a misnomer in most cases and the less misleading self poisoning replace it.


What do New Zealand general practitioners do in their offices?

A third person physician observer timed 16 New Zealand general practitioners during a work day during summer months. Usual office hours for the 16 physicians occupied over seven hours, during which time the patient list averaged 26 persons. Greatest time span, 27 percent, used in diagnosis, but substantial time was spent in health education, counseling and administration. The distribution of patients' problems was greatest in respiratory disease, followed by musculoskeletal, dermatologic and urogenital. There is considerable similarity to practice patterns of rural physicians in Missouri, USA.


Physiotherapy and the frozen shoulder: a comparative trial of ice and ultrasonic therapy.

In a prospective study ice and ultrasonic applications have been compared in a series of patients with the frozen shoulder syndrome. No significant advantage of one treatment over the other could be demonstrated. It would appear that both of these physiotherapy methods can shorten the painful stage of the condition and, in conjunction with specific exercises, hasten recovery of the range of shoulder movement.


Serum zinc levels in New Zealand children.

A report is made of serum zinc levels in 518 children with an age range of nine days to 17 years. There were significant differences in normal levels between fasting and non-fasting children, Europeans and Maoris, Europeans and Polynesians, and Indians and Polynesians. Lower levels were also found in those under the age of 13 but may be due to variation in serum proteins in these children who were all anaesthetised.


Benign self-limiting sarcoidosis with skin and joint involvement.

Three cases of sarcoidosis presenting with both erythema nodosum and erythema diffusion as well as with an arthritis and pulmonary involvement are described. The simultaneous occurrence of nodose and diffuse skin lesions would suggest that a clinical distinction between Lofgren's syndrome (Lofgren, 1953) and the syndrome described by Caplan, Katz, Rubenstein (1970) is not justified.


Histometric study of healing of periodontal tissues in rats after surgical injury. I. Design of a standardized surgical procedure.

A standardized surgical procedure in rats is described which allowed a histometric registration of the repair of cementum, periondontal ligaments and bone, and which eliminated the possibility of interference of pocket epithelium during the healing process. Using an extraoral incision, the lateral surface of the mandible was exposed, and a cavity was cut through the vestibular bone plate, the periodontal ligaments and into the root surface, using a specially designed handpiece and bur. This procedure was performed in 20 male Wistar rats, 9 weeks old. The observation period was 12 weeks. The systematic error, precision and interanimal variation were examined for histometric registrations of the repair of cementum, periodontal ligaments and bone. The precision of the histometric procedures was in this study found to be 5% or less for most of the different structures measured, and the estimated combined systematic errors probably less than 5% for cementum repair. The coefficient of variation in cementum repair among different rats was 24%. This variance has two components, the combined influence of systematic errors and precision of histologic and histometric procedures and the other the biological variance due to inter-animal differences. The precision and systematic errors probably do not account for more than one third of this variance, the remaining two thirds being inter-animal differences. These differences could not be completely controlled in this study in spite of the use of a controlled genetic strain of rats bred in litters of the same size under SPF conditions.


Histometric study of healing of periodontal tissues in rats after surgical injury. II. Healing events of alveolar bone, periodontal ligaments and cementum.

A standardized surgical periodontal injury was performed in 67 male Wistar rats 9 weeks of age. An osteotomy was made with a burr through the vestibular bone plate and the periodontal ligaments and into the superficial part of the root surface of the mandibular first molar. The rats were sacrificed with observation periods ranging from 1 day to 2 years postoperatively. A histometric registration was made of the repair of alveolar bone, periodontal ligaments and cementum. Bone repair was found after 7 days and included a bony alkalosis. On the period from 7 to 14 days after surgery this ankylosis was removed by resorption process. Argyrophilic reticulin fibers were present 3 to 5 days after surgery. Precollagen fibers were found after 4 days, whereas mature collagen fibers inserting in new formed cementum, was seen after 14 days. The number of collagen fibers increased substantially until 28 days after surgery. Initial cementum repair was found after 14 days and increased substantially with increasing observation periods tending to reach the thickness of the contralateral control tooth. Based on these findings a new concept of healing of the surgically injured periodontium in rats is presented. According to this the healing occurs in three successive phases. 1. Bone repair with formation of an ankylosis. 2. Repair of the periodontal ligament including removal of the established ankylosis. 3. Repair of Cementum.


Radiography of the temporomandibular joint utilizing oblique lateral transcranial projections. Comparison of information obtained with standardized technique and individualized technique.

An individualized technique for radiography of the temporomandibular joint utilizing oblique lateral transcranial projection is described. The patient is fixed in a cephalostat and the central ray directed through the horizontal long axis of the condyle. The information yielded when applying the individualized technique has been compared with that yielded from a standard technique and tomography. The plane of sectioning for tomography was oriented perpendicular to the horizontal long axis of the condyle. The material comprised radiographs from 30 joints. The condyle to fossa relation was studied as well as the occurence of structural changes. Neither the individualized technique nor the standardized technique gave an accurate picture of the condyle to fossa relation when compared to tomography. However, the individualized technique tended to give the same result as tomography more often. Tomography revealed the greatest number of structural changes (66) followed by individualized technique (47) and standard technique (19). Most changes were noted exclusively in one projection. A combination of individualized technique and tomography revealed more changes than a combination of tomography and standard technique.


Treatment of non-vital permanent incisors with calcium hydroxide. VI. A clinical, microbiological and radiological evaluation of treatment in one sitting of teeth with mature or immature root.

The purpose of the investigation was clinically, microbiologically and radiologically to assess the effect of calcium hydroxide as a temporary root-filling inserted in the same sitting as root canal debridement in non-vital permanent incisors with mature and immature root, infected or uninfected root canal and with or without radiologically demonstrated periapical bone changes. The material consisted of 141 teeth divided in 3 groups in which mechanical cleansing was accompanied by flushing with sterile saline and sodium hypochlorite solutions giving 0.5% or 5.0% active chlorine, respectively. Microbiological samples were taken from root canals after extirpation of necrotic pulp tissue, after completed cleansing of the root canal and 3 and 6 month after treatment. Results of treatment were evaluated from the radiographs taken before treatment and at the 3 and 6 month follow-ups. Complication, pain and an abscess, occurred in 2 cases, 2 and 5 days, respectively, after treatment. No statistical correlation between occurrence of samples that gave growth, taken from the root canals at 3 (8%) or 6-month control (9%) and 1) bacteriological status of the root canal prior to filling with calcium hydroxide, 2) the development of the root or 3) periapical healing at 3 or 6 month follow-up could be ascertained. Periapical bone healing at the end of 6-months observation period was noted in 61 teeth (46%), regression of periapical bone lesions in 64 (49%) and no periapical healing in 6 (5%). The only difference in healing pattern, statistically significant on 0.1% level, was found in the group of teeth flushed with 5.0% sodium hypochlorite. At 3 month control they showed percentually less cases with regression and more cases with no healing of periapical bone lesions than the teeth in the other two groups. It was concluded that treatment in one sitting can be done routinely, irrespective of the initial status, in all those cases where no other treatment is possible. If the periodontium or the periapical bone are injured during cleansing procedures or if necrotic rests are not pressed out through the apical foramen, no complications after treatment need to be feared.


Results of therapy in adult acute lymphocytic leukemia.

27 adult patients with acute lymphocytic leukemia have been analyzed. Complete remission was induced in 9 of 113 patients treated with prednisone +6-MP, 4 of 6 patients treated with prednisone + vincristine, and the one patient treated with a combination of prednisone, vincristine, 6-MP, and methotrexate. The median survival for these 20 patients was 11 months and the median duration of complete remission was 71/2 months. Two of these patients remain in complete remission at 8+ years. Our most recent regimen for remission induction, prednisone + vincristine + daunomycin, has produced complete remission in 7 of 7 patients, with a median duration of complete remission of 15 months. Three patients remain in their original complete remission from 16+ to 24+ months, and 5 of these 7 patients remain alive from 16+ to 24+ months.


[Spontaneous radial segmentation of the nuclei of myeloblasts in a case of chronic granulocytic leukemia during blastic crisis. an electromicroscopical study (author's transl)].

By means of electron microscopy, myeloblasts with spontaneous radial segmentation of the nuclei from a patient with micromegakaryotic chronic myelosis in the blastic crisis were analysed. The centriol was found in the center of the cells and microtubules were found between the segments of the nucleus. These observations support the hypothesis that spontaneous radial segmentations of the nuclei of tumor cells is associated with a contractile process of the microtubules with split the nucleus.


[Melkersson-Rosenthal syndrome].

The Melkersson-Rosenthal syndrome consists of recurrent facial edema, mainly localized to the lips, recurrent facial paralysis and plicated tongue. The edema eventually becomes permanent, and in combination with residues of facial paralysis, may bring about considerable disfigurement, thus necessitating medical or surgical intervention. The different elements of the triad often do not occur simultaneously, and two positive findings are often considered sufficient for the diagnosis. A case report of a 52 year old woman with lip edema and partial facial paralysis is presented. Antihistamines and corticosteroids systemically were found to have no influence on the edema, and ultimately plastic correction was resorted to, without any striking improvement of her condition.


[Trial of androgen therapy in the treatment of non-lymphoblastic acute leukemia. First results].

Addition of a daily dose of androgen in the form of 0.15 mg/kg of Stanzolol, given without interruption, gave an average survival of more than 4 years in patients suffering from granulocyte series acute leukaemias after complete remission was obtained. The simplicity of this treatment is apparent only from the appearance of marked manifestations of androgen impregnation in women from the 8th month of treatment onwards. These results, superior to those obtained up to the present time in the survival of myeloid leukaemias (non-lymphoblastic) were also better in terms of the stability and X "quality" of the remission in comparison to those obtained in acute lymphoblastic leukaemias. Confirmation of these results by controlled clinical trial will open up interesting perspectives, along side immunotherapy which remains of unproven effectiveness in myeloid leukaemias. The effectiveness of androgen stimulation of haematopoiesis as a stabilising factor of complete remissions in acute leukaemias has, in addition, interesting implications with regard to the theory of the leukaemic process.


Screening by negative chemical ionization mass spectrometry for environmental contamination with toxic residues: Application to human urines.

A screening procedure based on solvent extraction with minimal clean up followed by examination with negative chemical ionization mass spectrometry has been developed for organochlorine residues and related compounds with masses greater than 130 daltons. The procedure has been made quantitative for analysis of pentachlorophenol residues. A limited survey of human urines indicated extensive contamination of the subjects, studied at parts per billion and higher levels, with pentachlorophenol, 2, 4, 5-trichlorophenoxy acetic acid, other polychlorophenoxy acids, and numerous unknown compounds. In an attempt to define the source of the contamination, the screening procedure was applied to samples of environmental substrates. The results suggest the food chain as one significant source of the contamination. A preliminary study of organochlorine compounds in human seminal fluid showed pentachlorophenol and other organochlorine residues in every sample, a result which suggests that this area may merit additional monitoring.


Conformation of uncomplexed [Phe4, Val6] antamanide crystallized from nonpolar solvents.

[Phe4, Val6] antamanide, a synthetic, biologically active analog of the cyclic decapeptide antitoxin isolated from Amanita phalloides, has been crystallized from a mixture of n-hexane and methyl acetate, and its conformation has been established by the direct method of x-ray diffraction analysis, i.e., without the benefit of any heavy atom. The uncomplexed molecule contains a 2-fold rotation axis and cis peptide linkages between Pro2-Pro3 and Pro7-Pro8. Otherwise, its conformation differs extensively from that of the Na+[Phe4, Val6] antamanide-C2H5OH complex. The 30-membered ring is elongated and relatively planar as compared to the folded ring in the Na+ complex. The six NH groups are directed toward the anterior of the molecule. There is one pair of intra-molecular NH---O=C bonds of the 5 leads to 1 type containing a cis peptide unit. The other four NH groups participate in hydrogen bonds to three H2O sites in the interior of the molecule. The 10 hydrophobic side groups cover the bottom and surround the perimeter of the molecule with the phenyl groups in the four Phe residues folded against the molecule. The conformation found for [Phe4, Val6] antamanide crystallized from nonpolar solvents is different from any conformations proposed for antamanide in solution based on nuclear magnetic resonance data.


Structural and kinetic studies of lasalocid A (X537A) and its silver, sodium, and barium salts in nonpolar solvents.

The ionophore lasalocid A (X537A) and its metal salts have been investigated by high resolution (270 MHz and 360 MHz) proton nuclear magnetic resonance spectroscopy to obtain structural and kinetic information in nonpolar solution. The proton resonances were assigned from double resonance studies on lasalocid A and on its salts, homologs, isomers, and chemically modified derivatives. Studies of proton and carbon longitudinal relaxation time suggest that lasalocid A exists as a monomer, whereas the sodium and barium salts exist as dimers in nonpolar solvents. A study of the magnitude of the vicinal proton coupling constants and the chemical shifts and linewidths of the hydroxyl resonances suggest that the backbone conformation and intramolecular hydrogen bonds are similar for lasalocid A and its sodium and barium salts in nonpolar solvents. Nuclear magnetic resonance studies on the role of bound solvent molecules suggest a tightly bound water molecule in the barium complex dimer (crystallized from water-ethanol) and a weakly bound ethanol molecule in the lasalocid A monomer (crystallized from ethanol) in cyclohexane. The selective changes in proton chemical shift on complexation [where the polar faces of two lasalocid anions coordinate the metal cation(s) in nonpolar solvents have been analyzed in terms of the proximity of the resonances to the cation, their linkage to the coordinating oxygen atoms, and the magnetic anisotropy effects of the polar groups of one ligand on the resonances of its partner in the dimer. The nuclear magnetic resonance studies in solution are compared with earlier observations on lasalocid A and its salts in the crystalline state. Thus, the short Ag-C5 distance in the crystal structure of silver complex dimer is also observed in the solution structure. The kinetic parameters associated with the exchange between lasalocid A and its barium complex in chloroform have been measured from an analysis of the resonance line shapes as a function of temperature.


Localization of acetylcholine receptors during synaptogenesis in retina.

Nicotinic acetylcholine receptors are synthesized in chick embryo retina before synapses appear. Most of the receptors are found associated with neurites in the inner synaptic layer of the retina; later in development the receptors appear in the outer synaptic layer. Cells dissociated from retina and cultured in vitro form aggregates and also sort out into regions comprised either of neurites or cell bodies. Most of the nicotinic acetylcholine receptors of aggregates are found associated with neurite regions. The receptor distribution of cultured retina cells thus resembles that of intact retina.


Unequal accumulation of alpha- and beta-globin mRNA in erythropoietic mouse spleen.

Relative amounts and rates of synthesis of alpha- and beta-globin mRNAs were determined during splenic erythropoiesis in mice. At times after injection of mice with phenylhydrazine, alpha- and beta-globin mRNAs were separated by gel electrophoresis and quantitated by densitometric scanning of stained gels. At 66 hr after injection, the ratio of beta to alpha mRNA is about 1.2. By 138 hr, total globin mRNA is 5-fold greater in spleen cells, and the beta to alpha mRNA ratio approaches 2. This ratio remains around 1 in reticulocytes throughout this period. Analyses of globin products directed by these mRNAs from spleen cells and reticulocytes in the ascites cell-free system reflect the beta to alpha mRNA ratio observed by electrophoresis. Relative rates of synthesis of globin mRNAs were estimated after incubation of spleen cells with either [3H] uridine or [3H] adenosine. Although synthesis of both mRNAs is maximal at 114 hr and then declines sharply, beta mRNA is synthesized at a greater rate than alpha mRNA at every developmental stage. In contrast to the excess accumulation of beta mRNA in spleen cells, synthesis of alpha- and beta-globin chains remains balanced throughout erythroid development. These data suggest that during erythropoiesis in this system, equal synthesis of alpha and beta globin involves regulation at both transcriptional and post-transcriptional levels.


Opioid activity of a peptide, beta-lipotropin-(61-91), derived from beta-lipotropin.

The pituitary peptide beta-lipotropin displays essentially no opoid activity in a preparation of guinea pig ileum or in the opiate receptor binding assay. However, a fragment, beta-lipotropin-(61-91), with the enkephalin sequence (Tyr-Gly-Gly-Phe-Met) at its NH2-terminus, has typical opioid effects in these two assays.


Crystalline semisynthetic ribonuclease-S'.

Crystals of solid phase-derived semisynthetic ribonuclease-S' were prepared and compared with those for native ribonuclease-S' and -S. The semisynthetic species used was the noncovalent complex of synthetic fragment-(1-20), corresponding to residues 1 through 20 of bovine pancreatic ribonuclease-A (ribonucleate 3'-pyrimidino-oligonucleotidohydrolase, EC 3.1.4.22), and native ribonuclease-S-(21-124); the fragment containing residues 21 through 124 of ribonuclease-A. This semisynthetic complex was completely active enzymatically, was homogeneous as judged by polyacrylamide gel electrophoresis, and had no greater than trace amounts of excess ribonuclease-s(21-124) as judged by affinity chromatography. Crystallization of both semisynthetic and native ribonuclease-s' at pH 5.3 resulted in well-formed crystallseater than trace amounts of excess ribonuclease-S-T21-124) as judged by affinity chromatography. Crystallization of both semisynthetic and native ribonuclease-S' at pH 5.3 resulted in well-formed crystals with the symmetry of space group P3121 and unit cell dimensions a=b=44.82, c=97.3 A. This crystal form corresponds to the Y form of native ribonuclease-S previously reported [Wyckoff et al. (1967) J. Biol. Chem. 242, 3749-3753]. X-ray diffraction patterns of the crystals were indistinguishable, indicative of the structural identity of semisynthetic and native ribonuclease-S'.


Amino acid sequence at the FdUMP binding site of thymidylate synthetase.

Cyanogen bromide treatment of thymidylate synthetase of Lactobacillus casei, which had been converted to a ternary complex with [2-14c] FdUMP and 5,10-methylene-tetrahydrofolate followed by S-carboxymethylation, yielded at least four visible peptide bands, the largest with a molecular weight of about 13,000, on polyacrylamide gel electrophoresis in sodium dodecyl sulfate-urea. Identical results were obtained with enzyme that had all four of its cysteinyl residues S-carboxymethylated with iodo [I-14C] acetate in the absence of FdUMP and cofactor. In each case, only the second band from the top of the gel (CN2), with an approximate molecular weight of 10,000= was labeled. Analysis of CN2 that had been labeled with [2-14C] FdUMP and nonradioactive iodoacetate and of that labeled only with iodo[1-14C] acetate revealed that their amino-acid contents were almost identical except for the presence of two S-carboxymethyl (Cm)-cysteinyl residues in the latter peptide and only one in FdUMP-CN2. A nonapeptide was isolated from (Cm)2-CN2 after chymotrypsin digestion that contained the following sequence by dansyl-Edman analysis: Ala-Leu-Pro-Pro-[Cm-Cys]-His-Thr-Leu-Tyr. This peptide was found to be located on the NH2-terminal end of CN2. Automatic sequence analysis of the first 13 residues of (Cm)2-CN2 and of the FdUMP-containing CN2 yielded identical results except for the fifth, or cysteinyl, residue, which could not be identified in the latter peptide. These findings strongly suggest that FdUMP is linked to a cysteinyl residue in thymidylate synthetase that has been inactivated irreversibly by this nucleotide.


Guanidine-unfolded state of ribonuclease A contains both fast- and slow-refolding species.

The kinetics of the refolding reaction of ribonuclease A from high concentrations of guanidine hydrochloride or urea are biphasic, and show two refolding reactions whose rates differ 450-fold at pH 5.8 and 25 degrees. Measurements of cytidine 2'-phosphate binding during refolding, after stopped-flow dilution of guanidine hydrochloride (Gdn.HCl) or urea, show that functional bovine pancreatic ribonuclease A (RNase A; ribonucleate 3'-pyrimidino-oligonucleotidohydrolase, EC 3.1.4.22) is formed in both the fast and slow phases of the refolding process. We conclude that the guanidine-unfolded state of RNase A is an equilibrium mixture of fast- and slow-refolding species, as was found previously for the heat-unfolded state at low pH. The fraction of the fast-refolding species in guanidine or urea-unfolded RNase A is the same as that in the heat-unfolded protein at pH 2. Previous work has shown that the fast-refolding species disappears as the pH is raised from 3 to 5 for heat-unfolded RNase A. This pH effect is not present in refolding from concentrated Gdn.HCl solutions: the same proportion of the fast-refolding species is found from pH 2 to pH 6, and also from 2 M to 6 M Gdn.HCl at pH 5.8. We conclude that the same proportion of the fast-refolding species is present at equilibrium whenever the residual structure in unfolded RNase A is reduced to a low level, and that the structural difference between the fast-refolding and slow-refolding species of RNase A lies in the configuration of the random coil polypeptide chain.


Induction of a protein inhibitor to ornithine decarboxylase by the end products of its reaction.

Putrescine, the end-product of ornithine decarboxylase (ODC: L-ornithine carboxylyase, EC; 4.1.1.17) action, induces the synthesis of a protein(s), in L1210, neuroblastoma, and H-35 cells as well as in rat liver, which inhibits ODC activity. Spermidine and spermine, distal products of ODC activity, also induce the synthesis of a similar protein in H-35 cells. These ODC-inhibitors are heat-labile, trypsin-sensitive, and their induction is dependent upon protein synthesis. They have short half-lives which range from 18 to 66 min; these half-lives are similar to those of the ODC derived from the same source. They are noncompetitive inhibitors of ODC activity with an apparent molecular weight of 26,500. Each inhibitor crossreacts with the ODC's of the other cells and forms an enzyme-inhibitor complex which is stable during Sephadex chromatography; however, after treatment with ammonium sulfate, enzyme and inhibitor activities can be dissociated and recovered intact from the same column. We propose the name antizyme for proteins whose synthesis is induced by the proximal or distal products of the enzyme they inhibit.


Combined effects of dissociable and undissociable local anesthetics upon ATP-induced firefly bioluminescence.

Combined effects of two drugs present simultaneously are usually expressed as summation, synergism or potentiation, and antagonism. When the sum of the effects of each drug present separately equals the combined effect of the two drugs present simultaneously, the action is called additive or summation. However, the expected value of the sum of each effect of drugs present alone has not been well defined. In this report, the thearetical value of the expected sum of each effect of two inhibitors is given and a graphical method is presented to visualize summation, synergism, and antagonism. The inhibitory effects of a dissociable local anesthetic, tetracaine, and an undissociable local anesthetic, benzyl alcohol, upon a soluble firefly luminescent system were analyzed according to the above theory. The results clearly indicate that the action of these two classes of local anesthetics is pure additive or summation.


Solenoidal model for superstructure in chromatin.

Chromatin prepared by brief digestion of nuclei with micrococcal nuclease, and extracted in 0.2 mM EDTA, appears in the electron microscope as filaments of about 100 A diameter which coil loosely. In 0.2 mM Mg++ these "nucleofilaments" condense into a supercoil or solenoidal structure of pitch about 110 A corresponding to the diameter of a nucleofilament. It is proposed that the x-ray reflections at orders of 110 A observed in chromatin originate in the spacing between turns of the solenoid rather than that between nucleosomes along the nucleofilament. The solenoidal structure appears to need histone H1 for its stabilization. Under certain conditions, isolated nucleosomes can also aggregate into a similar structure. The solenoidal structure can be correlated with the "thread" of diameter about 300 A observed by other workers in nuclei.


Amino-acid sequence of tropomyosin-binding component of rabbit skeletal muscle troponin.

The complete amino-acid sequence of rabbit skeletal troponin-T is reported. The protein consists of a single polypeptide chain of 259 amino acids; it has an acetylated amino terminus and a molecular weight of 30,503. The sequence was determined by manual and/or automated Edman degradation techniques on the six fragments obtained after cleavage with cyanogen bromide. The larger fragments were further digested with trypsin, chymotrypsin, alpha-lytic protease, thermolysin, or pepsin to obtain smaller fragments suitable for manual sequencing. About 50% of the residues are charged at neutral pH with highly acidic amino-terminal (residues 1-39) and highly basic carboxyl-terminal regions (residues 221-259). Predictions of secondary structure indicate 37% helical content with two long sections (residues 80-102 and 122-146) in that portion of the molecule implicated in binding to tropomyocin. Two of the three phosphorylated sites in the molecule are located at serine-1 and serine-149 or -150. The sequence about the latter site resembles somewhat the phosphorylase kinase phosphorylation sites in phosphorylase alpha and troponin-I.


Sequential translation of nonstructural proteins in cells infected with a Semliki Forest virus mutant.

Four nonstructural proteins with apparent molecular weights of 70,000 (ns-70), 86,000 (ns-86), 78,000 (ns-78), and 60,000 (ns-60) were translated in cells infected with Semliki Forest virus ts-1 mutant and maintained at the restrictive temperature. After synchronization of the initiation of protein synthesis these proteins were synthesized in the above order, suggesting that they are translated as a polyprotein starting from one initiation site. Two short-lived intermediates with apparent molecular weights of 155,000 and 135,000 were regularly detected. The former is presumably the precursor of proteins ns-70 and ns-86 and the latter of ns-78 and ns-60. The sequence of the structural proteins in their polyprotein was confirmed to be capsid-envelope E-2 (and E-3)-envelope E-1, beginning from the NH2-terminal end. We conclude that in Semliki-Forest-virus-infected cells two polyproteins are synthesized, one for the structural (130,000 daltons), the other for the nonstructural proteins (close to 300,000 daltons).


Cooperative interactions of tetrahydrofolate with purified pig kidney serine transhydroxymethylase and loss of this cooperativity in L1210 tumors and in tissues of mice bearing these tumors.

Serine transhydroxymethylase (5,10-methylenetetrahydrofolate: glycine hydroxymethyl transferase, EC 2.1.2.1) purified 200-fold from pig kidneys showed cooperative interactions with tetrahydrofolate with a Hill coefficient (n value) of 3.9 and a substrate concentration at 50% of maximum velocity, the S(0.5) value, of 0.5 mM. The enzyme in mouse liver and kidney homogenates also showed cooperative interactions with tetrahydrofolate. However, the enzyme obtained from L1210 solid tumors of mice, and from livers and kidneys of mice inoculated with L1210 cells exhibited hyperbolic saturation kinetics and gave a Michaelis constant, Km, value of 0.5 mM for tetrahydrofolate. The interaction of serine with the enzyme from pig kidney, from tissues of normal or tumor-bearing mice, or from L1210 tumors was hyperbolic with a Km of 0.9 mM. The specific activities of the enzyme in the L1210 tumor and in mouse liver were 10-fold higher than in pig or mouse kidney. There was no significant change in the levels of the enzyme in mouse liver and kidney on inoculation with L1210 cells. These results suggest that a tumor can bring about biochemical changes in tissues that are distal to the tumor.


Homologous inhibitors from potato tubers of serine endopeptidases and metallocarboxypeptidases.

A potent polypeptide inhibitor of chymotrypsin has been purified from Russett Burbank potatoes. The inhibitor has no effect on bovine carboxypeptidases A or B but exhibits homology with a carboxypeptidase inhibitor that is also present in potato tubers. The chymotrypsin inhibitor has a molecular weight of approximately 5400 as estimated by gel filtration, amino acid analysis, and titration with chymotrypsin. The polypeptide chain consists of 49 amino acid residues, of which six are half-cystine, forming three disulfide bonds. Its size is similar to that of the carboxypeptidase inhibitor, which contains 39 amino acid residues and also has three disulfide bridges. In immunological double diffusion assays, the chymotrypsin inhibitor and the carboxypeptidase inhibitor do not crossreact; however, automatic Edman degradation of reduced and alkylated derivatives of the chymotrypsin inhibitor, yielding a partial sequence of 18 amino acid residues at the NH2-terminus, reveals a similarity in sequence to that of the carboxypeptidase inhibitor. Thus, inhibitors directed toward two distinct classes of proteases, the serine endopeptidases and the metallocarboxypeptidases, appear to have evolved from a common ancestor.


Responses of Phycomyces indicating optical excitation of the lowest triplet state of riboflavin.

Phototropic and light growth responses of the sporangiophore of Phycomyces have been elicited using tunable laser stimulation from 575 to 630 nm. The growth response shows additional components of the action spectrum with a sharp peak at 595 nm, a sharp cut-off at 585 nm, and a tail extending beyond 630 nm. The integral over the electronic transition (f-value) is 1.5 X 10(-9) times that at 455 nm. These parameters indicate a direct transition from the ground state to the lowest triplet state of riboflavin.


Conformational flexibility and protein folding: rigid structural fragments connected by flexible joints in subtilisin BPN.

Conformational energy calculations are used to analyze the interactions of structural substructures in subtilisin BPN. These substructures are kept fixed or "rigid" so that the only variables in the calculations are the backbone segments that separate them. The flexible segments are assumed to be free turns. Using this representation of the protein it is possible to predict both a likely order of events along a folding pathway and preferred modes of conformational changes of the native protein. Moreover, when the native structure has been perturbed by moving the substructures apart, it is possible to assess the range of interactions that return the protein, upon energy minimization, to its original conformation. These results suggest an approach to the folding problem based on the piecemeal formation of tertiary structure from smaller prefolded fragments.


Size of primary transcripts in Ehrlich ascites cells as measured by tetraphosphate determination.

A method for the quantitation of 5"-tetraphosphate ends in 32P-labeled RNA has been developed. The tetraphosphate content of different RNA fractions obtained from Ehrlich ascites cells labeled with 32P for different lengths of time has been determined. Ribosomal RNA and poly(U)-binding RNA, labeled for long periods, (mRNA) lack 5'-terminal tetraphosphate. 5S RNA, pulse labeled 4-5S RNA, and poly(U)-binding hnRNA (heterogeneous nuclear RNA) do contain tetraphosphate. From the amount of the tetraphosphate, molecular weight data can be calculated for these RNA fractions which agree with independent determinations by denaturing gel electrophoresis. The results demonstrate that the majority of the poly(A) containing hnRNA molecules are small (less than 28S) and contain the tetraphosphate of the primary transcript. Therefore, they do not originate from the 3'-end of large molecules by processing events.


Sensitivity of calcium binding in cerebral tissue to weak environmental electric fields oscillating at low frequency.

Weak sinusoidal electric fields modify the calcium efflux from freshly isolated chick and cat cerebral tissues bathed in Ringer's solution, at 36 degrees. Following incubation (30 min) with radioactive calcium (45Ca2+), each sample, immersed in fresh solution, was exposed for 20 min to fields at 1, 6, 16, 32, or 75 Hz, with electric gradients of 5, 10, 56, and 100 V/m in air. 45Ca2+ efflux in the solution was then measured in 0.2 ml aliquots and compared with efflux from unexposed control samples. Field exposures resulted in a general trend toward a reduction in the release of the preincubated 45Ca2+. Both frequency and amplitude sensitivities were observed. Maximum decreases occurred at 6 and 16 Hz (12-15%). Thresholds were around 10 and 56 V/m for chick and cat tissues, respectively. Similar but nonsignificant trends occurred during other field exposures. All results were statistically compared with matched samples of controls. Tissue gradients could not be measured, but estimates were of the order of 0.1 muV/cm. The susceptibility of the electrochemical equilibrium in the neuronal membrane to small extracellular perturbations is discussed and a possible role for weak intrinsic cerebral fields in neuronal excitability is suggested.


In vitro derived mouse A9 cell clones differing in malignancy: analysis by somatic cell hybridization with YACIR lymphoma cell clones.

A culture of mouse A9 cells yielded clones of both high and low malignancy. Confirmation of the differences in malignancy of the clones was obtained after their hybridization with subclones of YACIR a lymphoma induced by Moloney leukemia virus. Low malignant A9 clones suppressed the malignancy of the YACIR partner in the hybrids. Hybrids between a highly malignant A9 cline and two different clones of YACIR "complemented" to yield hybrids of reduced malignancy. No correlation was formed between the malignancy of a particular A9 clone and its ability to produce a focus-forming mouse C-type virus.


Protein synthesis and actin heterogeneity in calf muscle cells in culture.

Pulse-labeled cytoplasmic proteins from cultured fetal calf-muscle cells at various stages of development were analyzed by one- and two-dimensional gel electrophoresis. The high resolution of the two-dimensional technique allows the determination of those protein species that begin to be synthesized after cell fusion. In addition, actin has been found to exist in three forms possessing similar biochemical properties and identical molecular weights but having slightly different isoelectric points. Two of the forms are found in prefusion dividing myoblasts and also in cultured kidney cells. The third form is the only one found in fetal muscle tissue and is predominant in cultures of fused muscle cells. Thus, it would seem that actin can exist in several isozymic forms of which one is specific to fused muscle tissue.


Selenium is an essential trace nutrient for growth of WI-38 diploid human fibroblasts.

The trace element selenium is essential for clonal growth of diploid fibroblasts from human fetal lung (WI-38) in media containing small amounts of serum protein. Maximum growth stimulation is obtained when 30 nM neutralized selenious acid is added to a synthetic medium containing 1.5 mg/ml of dialyzed fetal bovine serum protein (equivalent to a 3% serum concentration). Serum appears to be a source of selenium in most culture media, since higher concentrations of serum protein or whole serum mask the selenium requirement of WI-38 cells. Selenium is also required by a Chinese hamster cell line that can be grown in a protein-free synthetic culture medium.


Induction of maturation (meiosis) in small Xenopus laevis oocytes by injection of maturation promoting factor.

Treatment of small-(stage III) or medium-sized (stage IV) Xenopus laevis oocytes with progesterone, human chorionic gonadotropin, or para-hydroxymercuriphenylsulfonate does not induce maturation. Only the full-grown oocytes (stage VI) undergo maturation when treated with either one of these three substances. In contrast, injection of maturation promoting factor into oocytes of stages III-VI invariably leads to chromosome condensation and germinal vesicle breakdown. No maturation spindle is found in oocytes smaller than 0.9 nm in diameter, and the nuclear sap does not mix with the cytoplasm in the smallest (0.45-0.55 mm in diameter) oocytes. In oocytes of 0.9 mm, maturation is identical to that of full-grown oocytes, except that the maturation spindle does not reach the cortex of the oocyte. Progesterone increases protein synthesis in medium-sized (0.8 mm in diameter) oocytes without inducing meiosis. It has little or no effect on protein synthesis in smaller oocytes.


Stimulation of fetal hemoglobin synthesis in bone marrow cultures from adult individuals.

The regulation of fetal hemoglobin in adult erythroid cells was investigated with bone marrow cultures. Fetal hemoglobin (Hb F) was identified in individual erythroid colonies with fluorescent antibodies against Hb F and synthesis of gamma chains was determined with analyses of radioactive globins. The appearance of fetal hemoglobin in erythroid colonies was clonal. All the cells of the Hb F synthesizing colonies contained fetal hemoglobin. The frequency of erythroid colonies showing Hb F was higher than expected compared to the frequency of Hb F containing cells in the blood. Production of Hb F in culture, as shown by analysis of the radioactive globins, was 5 to 14 times higher than baseline Hb F synthesis. These results suggest that the ability for gamma chain synthesis in erythroid cells is determined at or above the level of the precursor cell from which the erythroid colonies, in vitro, derive (probably an erythropoietin responsive stem cell), and that stimulation of fetal hemoglobin synthesis in adult erythroid cells is possible.


Mapping of adenovirus messenger RNA by electron microscopy.

Late adenovirus messenger RNA was annealed to complementary regions of partially melted viral double-stranded DNA. The RNA. DNA hybrid regions within the DNA molecules were visualized as loops in the electron microscope. Loops occurred at several regions of the DNA, most frequently, however, at a location near the center of the molecule. This hybridization technique appears well suited for an accurate mapping of messenger RNA, as well as for studies of RNA processing.


Isolation from patients with breast cancer of antibodies specific for antigens associated with breast cancer and other malignant diseases.

Antibodies were specifically purified, with the use of pleural fluid antigens entrapped in a polyacrylamide gel as immunoadsorbent, from a pool of sera from patients with breast cancer and from the pleural effusion of an individual patient. The purified antibodies were radioiodinated and tested for capacity to bind to the pleural fluid adsorbent. Binding of the radiolabeled antibodies was inhibited by many of the sera from women with breast cancer to a much greater extent than by sera of healthy women. With purified antibodies originating from the serum, 52% of the breast cancer sera and 16% of sera from patients with other malignancies were more inhibitory than were 95% of the sera of healthy women. In the tests with antibodies eluted from pleural fluid, 24% of the breast cancer sera and 25% of the sera from patients with other neoplastic diseases were more inhibitory than were 95% of the normal sera. We concluded that sera of patients with breast cancer may contain antibodies against antigens associated with breast cancer as well as additional antibodies against antigens that are also found in patients with other neoplastic diseases or in normal individuals. Individual breast cancer sera may inhibit binding of the labeled antibodies against breast cancer owing to their content of antibodies, antigen(s), or both.


Rheumatoid factor and tumor-host interaction.

In this survey for rheumatoid factor (RF) seropositivity on patients with neoplasms, an 85% rate of positive screening tests was recorded under certain circumstances. This high rate of RF seropositivity occurred after irradiation and/or chemotherapy of breast and lung cancers. Treated patients with breast cancers who had no evidence of residual tumor had an 89% rate of positive RF tests. Conversely, the incidence of RF seropositivity was low among untreated patients with similar tumors and treated patients with glioblastomas or multiple myeloma. The administration of cytotoxic drugs (e.g., azathiprene) was not itself associated with RF production even in renal allograft recipients. The data indicate that RF production occurs frequently after therapy of certain tumors and suggest that in these circumstances RF may be an expression of tumor-host interaction.


Mechanism of human lymphocyte stimulation by concanavalin A: role of valence and surface binding sites.

A monovalent form of concanavalin A (m-Con A) has been prepared to determine the importance of valence for human lymphocyte surface binding and subsequent lymphocyte stimulation as measured by blast transformation and cytotoxicity. Concanavalin A (Con A) was fragmented by a proteolytic process and the m-Con A) derivative was isolated by elution with an ascending D-glucose gradient on a Sephadex G-200 column. The molecular weight of m-Con A was 18,000 by sodium dodecyl sulfate-polyacrylamide electrophoresis. Equilibrium dialysis with alpha-methyl D-glucoside and subsequent Scatchard plot analysis revealed an association constant (Ka) of 1.2 X 10(3) liters/mol and a valence of 1.1. Incubation of lymphocytes with 125I-labeled m-Con A demonstrated surface binding at 1.21 X 10(6) molecules per cell, which was comparable to the binding of [3H] Con A (1.02 X 10(6) molecules per cell). However, in contrast to the intact lectin, m-Con A had a markedly reduced capacity to agglutinate rabbit erythrocytes and human lymphocytes and did not stimulate lymphocyte blast transformation or cytotoxicity at 1 and 10 mug/ml. Finally, pretreatment of lymphocytes with m-Con A blocked blast transformation induced by Con A. These observations demonstrate that m-Con A binds to lymphocyte surface receptors but does not stimulate blast transformation or cytotoxicity, suggesting that Con A must bridge binding sites on the lymphocyte surface to induce lymphocyte activation.


Immunological cross-reactivity of antibodies to a synthetic undecapeptide analogous to the amino terminal segment of carcinoembryonic antigen, with the intact protein and with human sera.

A peptide corresponding to the 11 amino acid residues of the NH2-terminal portion in the sequence of carcinoembryonic antigen(CNTHETIC CEA(1-11) peptide was attached by means of a water-soluble carbodiimide reagent to multichain poly(DL-alanine( as well as to bovine serum albumin. Both macromolecular conjugates provoked in rabbit anti-CEA(1-11) peptide antibodies. The specificity of this immunological system and the crossreactivity between the peptide and intact CEA were investigated by two methods--passive hemagglutination and modified bacteriophage inactivation. Hemmagglutination experiments showed that not only anti-CEA(1-11) sera, but also anti-CEA sera, agglutinated CEA(1-11)-coated sheep erythrocytes, and both these reactions were inhibited with CEA(1-11) peptide. In experiments with the chemically modified bacteriophage technique CEA(1-11)-coated phase was efficiently inactivated with antisera against the CEA(1-11) conjugates, and the inactivation reaction could be totally inhibited with the free peptide. The semipure CEA, but not the pure protein, could also inhibit the phage inactivation, even though less efficiently. On the basis of the above results, sera of some cancer patients were tested for their capacity to inhibit the inactivation of CEA(1-11)-coated phage by means of anti-CEA(1-11) antiserum. The results indicate that sera from a large proportion of patients with adenocarcinomas of the digestive tract, pancreas, and breast are capable of inhibiting the above inactivation, whereas most normal sera do not inhibit.


Soluble dextran-hemoglobin complex as a potential blood substitute.

A complex between soluble dextran and human hemoglobin has been synthesized by two different methods. In the alkylation method, hemoglobin was allowed to react with bromoacetyl groups incorporated into the dextran; the yield of the complex was about 80% in terms of the hemoglobin used. In the dialdehyde method, hemoglobin was allowed to react with dialdehyde groups on the dextran generated by periodate oxidation; the yield of the complex was about 60%. Both soluble dextran-hemoglobin complexes could bind and release oxygen reversibly, but the oxygen-binding curves were shifted to the left relative to that of free hemoglobin. In the rabbit, the complex obtained by the alkylation method was excreted by the kidneys and cleared from the circulation much more slowly than free hemoglobin.


Antibodies as a means of isolating and characterizing biologically active substances: presence of a non-peptide, morphine-like compound in the central nervous system.

Antibodies generated against small molecular weight substances such as drugs are being used to isolate and characterize biologically active agonists. A morphine-like compound can be extracted from brain of various species which has determinant groups that are recognized by specific morphine antibodies. It has a regional distribution which can be quantitated as immuno-equivalents. Immunological, chemical and chromatographic tests show great similatiries of the compound to morphine. This morphine-like compound has biological activity as it inhibits the electrically induced contractions both of the guinea pig ileum and mouse vas deferens but the inhibition is not reversed by naloxone or naltrexone.


Mapping of the influenza virus genome: identification of the hemagglutinin and the neuraminidase genes.

Polyacrylamide gel electrophoresis of the RNA of influenza A/PR/8/34 (H0N1) and A/Hong Kong/8/68 (H3N2) viruses and recombinant viruses derived from them revealed that each contains eight RNA segments, the fourth of which codes for hemagglutinin. (The largest RNA of the segmented genome is counted as band 1.) The neuraminidase gene was identified as the sixth segment in the RNA pattern of influenza A/PR8 virus and as the fifth segment of A/Hong Kong virus. The molecular weights of the RNAs for the hemagglutinin and the neuraminidase genes lie in the range of 600,000-700,000.


beta-Lipotropin as a prohormone for the morphinomimetic peptides endorphins and enkephalins.

The hypophysial homomeric peptide beta-lipotropin (beta-LPH-[1-91]) has no morphinomomimetic activity in a bioassay (myenteric plexus-longitudinal muscle of the guinea pig's ileum) or binding assays with stereospecific opiate-receptors of rat brain synaptosome preparations. Incubating beta-LPH-[1-91] at neutral pH with the supernatant aqueous extracts of rat brain generates (fragments of beta-LPH with) morphinomimetic activity in the same assay systems. These results are related to the recently recognized structural relationships between beta-LPH, the newly isolated peptides met-enkephalin (beta-LPH-[61-65]) and alpha-endorphin (beta-LPH-[61-76]) and also to the biologically active fragments of analogs: beta-LPH-[61-64], beta-LPH-[61-65[-NH2, (Met(O)65)-BETA-LPH-[61-65], beta-LPH-[61-69], and beta-LPH-[61-69]. Enzymatic biogenesis of these morphinomimetic peptides would preclude localizing them as such in cellular or subcellular elements with currently available methodology.


The role of radiation therapy in the treatment of osteosarcoma.

Seventy patients received primary surgical or radiotherapeutic treatment for osteosarcoma at the University of California-San Francisco between 1950 and 1974. Twenty-two received surgery alone and 27 received preoperative irradiation followed at varying intervals by surgery. Twenty-one patients received irradiation alone because of tumor location, refusal of recommended surgery, or development of metastatic disease. The results are analyzed and the role of radiation therapy in the primary management of osteosarcoma is discussed. The authors believe that irradiation without chemotherapy offers little in terms of palliation or enhanced survival in patients who are able to undergo radical surgery.


Multiple chromosomal associations and paracentromeric region instability in a case of acute leukemia.

A case of acute myelomonocytic leukema is described, which was characterized cytogenetically by the presence of centromeric elongations, somatic crossovers, selective endoreduplication figures, and multiple chromosomal clusters. The demonstration of these phenomena by selective staining techniques for the chromosome bands (Q, C, G and S) and the nucleolar areas (acridine-orange, amido black B 10) raises some biological aspects involved in the proliferation of leukemic cells, such as nucleolar persistance during the metaphase and the non-separation of chromatids in the clusters during the anaphase. These structural abnormalities may represent the background for the explanation of the appearance of subclones in neoplastic disorders.


Serum iron increase as a measure of iron absorption - studies on the correlation with total absorption.

The correlation between the magnitude of the increase of serum iron after an oral dose of iron and the total absorption of iron was studied in 51 healthy subjects and 10 patients with iron deficiency anaemia. 59Fe-labelled solutions of ferrous sulphate (25-100 mg iron) were administered to the fasting subjects. The serum iron concentration was followed for 4-6 hours and the absorption was measured in a whole-body counter. Good correlation was found between the maximal increase of serum iron and the total amount of iron absorbed after a dose of iron given as a solution. The serum iron method may be used for comparisons of the absorbability of different doses of iron by performing cross-over studies in groups of subjects. However, it was found that in an individual subject the serum iron method could not be used to determine the amount of iron absorbed from an oral dose of iron.


Absorption from different types of iron tablets - correlation between serum iron increase in total absorption of iron.

The absorption of iron from three different iron tablets - rapidly-disintegrating ferrous sulphate and ferrous carbonate tablets and slow-release ferrous sulphate tablets - was studied in healthy subjects with a serum iron technique and by whole-body counter measurements. A solution of ferrous sulphate was used as a reference. There were no differences in the absorption from the ferrous sulphate preparations, but the ferrous carbonate tablets were less well absorbed. Good correlation was found between the maximal serum iron response and the total absorption of iron and it was concluded that serum iron studies may be used for semiquantitative measurements of iron absorption in comparative studies on different iron preparations.


Influence of a liquid antacid on the absorption of different iron salts.

The influence of a liquid antacid on the absorption of iron from iron tablets was studied in healthy volunteers using a serum iron technique. Tablets containing ferrous salts (carbonate, fumarate, sulphate) with different in-vitro dissolution properties were studied. Co-administration of the antacid and the iron tablets resulted in a marked reduction of the absorption of iron from all preparations studied. The lowest absorption was found when ferrous carbonate was given with the antacid. This was ascribed to the fact that ferrous carbonate is almost completely insoluble at pH-levels near neutral. This was confirmed in a series of patients with achlorhydria in which the absorption from ferrous carbonate was virtually nil. It was concluded that iron tablets and antacid preparations should not be administered at the same time and that ferrous carbonate tablets should not be used in patients with achlorhydria.


Influence of food on the absorption from different types of iron tablets.

The influence of a standardized breakfast meal on the absorption of iron from two types of iron tablets - rapidly-disintegrating and slow-release tablets - was studied in 24 healthy subjects, using a serum iron technique and whole-body counter measurements. The meal markedly reduced the absorption of iron from both types of tablets. An 8-fold reduction was found with the rapidly-disintegrating tablets and a 3-fold reduction with the slow-release tablets. Under fasting conditions the rapidly-disintegrating tablets were significantly better absorbed than the slow-release tablets, whereas the reverse was found when the tablets were given with the meal.


Absorption of iron from slow-release and rapidly-disintegrating tablets - a comparative study in normal subjects, blood donors and subjects with iron deficiency anaemia.

The absorption of iron from slow-release and rapidly-disintegrating ferrous sulphate tablets has been compared using a double radioiron isotope technique. The studies were performed in 15 normal subjects, 20 blood donors and 10 patients with iron deficiency anaemia. The preparations containing 100 mg of ferrous iron were given twice daily on alternate days for 10 days. The absorption differed significantly between the three groups of subjects, being highest in the anaemic patients and lowest in the normal subjects. In all groups, significantly more iron was absorbed from the slow-release tablets compared to the rapidly-disintegrating tablets. The mean absorption ratios were 1.3 in both normal subjects and blood donors. In patients with iron deficiency anaemia the ratio was 1.4.


Blast crisis of chronic myeloid leukaemia: the effect of intensive chemotherapy.

24 patients with Philadelphia chromosome positive chronic myeloid leukaemia (CML) in blast crisis were treated with intensive chemotherapy. 16 patients showed either partial or complete response to this treatment, but median survival remained short (13 weeks), and much of this time was spent in hospital. These results were not significantly better than those obtained by others using vincristine and prednisolone alone, and this combination of drugs can often be given on an outpatient basis. It is concluded that until more effective intensive therapy becomes available patients in CML blast crisis should be managed in such a way that the quality of life is not impaired; and that at present vincristine and prednisolone appears to be the most impaired; and that at present vincristine and prednisolone appears to be the most appropriate initial treatment, though even this is far from satisfactory.


The penetration of trimethoprim into seminal fluid and serum.

A series of determinations of the concentrations of trimethoprim in seminal fluid and serum was made during the treatment of 18 men. Trimethoprim could be demonstrated in an average concentration in serum of 1.8-2.0 mug per millilitre and in seminal fluid of between 1.0 and 1.5 mug per millilitre. At the same time it was demonstrated that the combination of trimethoprim and sulphamethoxazole (co-trimoxazole) has no effect on genital mycoplasma species.


Co-trimoxazole in the long-term treatment of pyelonephritis with normal and impaired renal function.

The effect of co-trimoxazole was studied in 29 cases of acute or chronic pyelonephritis. The therapy produced the desired effect in 22 out of 23 cases and in the 6 cases treated prophylactically. We used a special dosage schedule for the combination of sulphamethoxazole and trimethoprim in the treatment of patients with normal renal function and with varying degrees of renal impairment. The results show that the plasma concentrations were at satisfactory therapeutic levels, with no accumulation of the three substances and without causing any toxic side effects, even in patients with severe impairment of the renal function. The mean duration of the treatment was 12.3 months. The material is, however, not largelu enough to warrant the recommendation of standardized treatment according to the schedule described, in spite of the favourable experience gained. It does, however, permit us to recommend that the plasma concentrations should be determined, particularly of the total sulphamethoxazole in patients with severely impaired renal function.