Recovery of tectal nicotinic-cholinergic receptor sites during optic nerve degeneration in goldfish. The concentration of cholinergic nicotinic-like sites as measured by alphabungarotoxin (alphaBuTX) binding, decreased in the goldfish (Carassius auratus) optic tectum after optic nerve disconnection. Initially, the rate of loss of sites is greater than the rate of tissue or protein degradation in experiments where disconnection was achieved either by unilateral optic nerve crush or by enucleation of one eye. When the crushed optic nerve is allowed to regenerate and form behaviorally potent connections, the number and concentration of these sites appears restored. Pharmacological studies indicate that the alphaButTX binding site in the goldfish optic tectum has a drug binding profile similar to that seen at central or peripheral alphaBuTX sites in other species.


Pulvinar and lateral geniculate neuronal activity in the cat during operantly conditioned appetitive behavior. Cats were trained to press a lever for 0.5--1.0 ml of milk reward both in the presence and absence of ambient light. 'Floating' microelectrodes were implanted in the pulvinar and lateral geniculate nuclei to allow chronic recording from single or multiple neurons over a period of 1--16 days. Thirty-one of 36 pulvinar neurons (86%) showed a consistent periodicity in the firing rate that was clearly related to certain phases of operant behavior. In the presence of ambient light, the firing rate was maximal shortly prior to or immediately after a rewarded bar press, particularly during the animal's initial contact with reward (the first lap). This burst of activity was followed by an abrupt and strong inhibition of firing which was, in most instances, associated with high voltage slow wave (6--9 c/sec) postreinforcement synchronization (PRS) of the electroencephalogram over the visual and association cortices as well as in the pulvinar. Postreinforcement inhibition of neuronal activity was also observed in the lateral geniculate. Neuronal firing in both nuclei dropped significantly below the respective mean rate observed for each nucleus during relaxed wakefulness. In the dark, however, the reward-induced inhibition was abolished in both nuclei. Moreover, some pulvinar neurons in the dark showed a conspicuous and significant increase in firing during the consummatory response (dark reversal pattern). These findings suggest that a transient but powerful inhibitory process triggered by positive reinforcement depends on visual input. Firing patterns of some pulvinar neurons were noted to be entrained not only by the rhythm of PRS oscillations (inhibitory phasing and postinhibitory discharges) but also by the much slower 3--4 c/sec rhythm of lapping and licking. Furthermore, the patterns of entrainment by initial lapping appear to be different from those observed during the later phase of the consummatory response, thus indicating that the pulvinar may receive information regarding the content of the delivery cup. No entrainment of neuronal activity was observed in the lateral geniculate. These findings reveal remarkable plasticity in the pulvinar neurons that is consistent with their role in polysensory integration.


Fuchs' heterochromic cyclitis associated with retinitis pigmentosa: a family study. To determine the hereditary, clinical histopathological aspects of the association between Fuchs' heterochromic cyclitis (FHC) and retinitis pigmentosa (RP), the family of a patient with FHC and bilateral RP was studied genealogically, ophthalmologically and immunogenetically. The oldest brother and the youngest sister of the proband had bilateral RP and glaucoma which in the brother lead to enucleation of an eye which was studied histologically. The proband, his brother with RP, and 2 of their healthy siblings were homozygous for the haplotype A3, B7, w6. The parents of the siblings were healthy, and the pedigree showed much parental consanguinity and indicated autosomal recessive inheritance of RP.


Lacrimal sac calculi.

Lacrimal sac calculi. Analysis of dacryoliths removed at operation showed that most consisted of calcium. One consisted of ammonium and could be directly related to the presence of bacteria. Tear samples obtained from 14 patients with dacryoliths were compared with seven normal patients with regard to calcium, phosphorus, and uric acid concentrations, tear to serum calcium ratios, and calcium-phosphate products. There was no significant difference between the two groups and thus no evidence that dacryoliths from because of abnormal tear electrolytes. We believe that dacryolith formation results from chronic obstruction and inflammation of the sac causing a build up of various electrolytes, particularly calcium. The preoperative dacryocystograms showed either a filling defect or apparent displacement of the lacrimal sac.


Rat retinal blood vessels.

Rat retinal blood vessels. The largest arteries in the rat retina are the arterioles in the nerve fiber layer adjacent to the optic papilla. They are 50 to 100 micrometer in diameter, have an incomplete internal elastic lamina and usually a single layer of smooth muscle. Smaller arterioles of 10 to 50 micrometer have no internal elastic lamina and the media is formed by one or two layers of slender smooth muscle cells. In these vessels, myoendothelial junctions and close contact areas between smooth muscle cells are numerous. Capillaries are present in all layers of the rat retina and from plexuses in the nerve fiber, outer plexiform and exterior part of the inner plexiform layers. In nearly all capillaries, pericytes and their processes from a single layer external to the endothelium with numerous contact points or zones between endothelium and pericytes without any intervening basal laminar material. Areas of close contact between adjacent pericyte processes are frequent. The possible functions of myoendothelial junctions and pericyte-endothelial contacts in relation to vessel tone, mechanical stabilization and metabolic exchange are discussed.


Congenital miosis.

Congenital miosis. The inheritance of congenital miosis in 2 pedigrees is described. The inheritance was autosomal dominant in one and autosomal recessive in the other. The pupils were 0.5-2.5 mm diameter, reacted normally to light and accommodation but dilated poorly with mydriatics. In both pedigrees, the affected members had enlarged corneas. The five members in the autosomal dominant family were myopic and had translucent peripheral irides. Iridodonesis was presented in both members of the autosomal recessive pedigree. The embryology and innervation of the iris muscles together with the ocular abnormalities and syndromes associated with congenital miosis are reviewed.


Bonnet-Dechaume-Blanc syndrome.

Bonnet-Dechaume-Blanc syndrome. A 16 year old girl presenting the visual disturbances was found to have a right retinal arteriovenous communication. The provisional diagnosis of Bonnet-Dechaume-Blanc syndrome was confirmed by angiographic demonstration of ipsilateral orbital and intracranital arteriovenous malformations.


A spectacle device to treat facial nerve paralysis. We describe a new method to prevent ocular complications in facial nerve palsy. A device mounted on a spectacle frame senses the blink of the normal eye and closes the lids of the paretic eye by levers which rest on the upper and lower lids. It provides a physiologic replacement of function and prevents corneal exposure.


The Panisset-Pop syndrome.

The Panisset-Pop syndrome. The authors describe a syndrome occurring in people learning fundus photography. This entity comprises the following: decrease of vision due to accomodative spasm, signs of anxiety and the finding of an improperly set ocular on the instrument used by the trainee.


Antihormone treatment of stage IV breast cancer. The antiestrogen Tamoxifen (T), given orally to 113 patients with stage IV breast cancer, induced objective remission in 50%. Duration of remission in the first 39 patients, with minimum 27 months follow up, is 18 + months; these results are equal to those of surgical hypophysectomy. T prolonged survival in responders. Older age, previous response to endocrine therapy and positive estrogen receptors predicted response to T. T was effective in hypophysectomized patients in whom serum growth hormone and prolactin were undetectable, but serum ostrogens were present in low amount, suggesting a direct stimulatory effect of estrogens at the tumor level. Hypophysectomy induced further palliation after treatment with T, indicating that pituitary hormones may also play a role in the growth of some human breast cancers. Side effects from T were minimal. T is the initial treatment of choice for postmenopausal women with hormone responsive stage IV breast cancer.


Further observations on the inhibition of tumor growth by C. parvum with cyclophosphamide. VII. Effect of treatment prior to primary tumor removal on the growth of distant tumor. The present investigations were directed toward determining whether primary tumor manipulation prior to its removal is advantageous for the control of metastases and survival. Studies were carried out to ascertain whether 1) there is justification for delaying surgical removal of a primary tumor to permit preoperative administration of cyclophosphamide (CY) and/or C. parvum (CP) and 2) there is an advantage to administering the immunotherapy directly into a primary tumor. After operation, in all investigations, systemic CP and CY was used. Despite the putative similarity of animals, tumors and treatment regimens there was marked variation in response of tumors to therapy. No benefit was derived from administering preoperative immunotherapy alone. When operation was delayed to employ systemic immuno-chemotherapy, a slight improvement in the control of distant tumor was noted. The employment of preoperative intratumor immunotherapy led to a greater prolongation of survival and more inhibition of distant tumor growth than did immediate primary tumor removal or the use of preoperative systemic immunotherapy. The results suggest that there may be an advantage to delaying removal of a primary tumor so that it may be employed in therapeutic strategies directed toward control of metastatic disease.


Cervix cancer: treatment with combination vincristine and high doses of methotrexate. Patients with carcinoma of the cervix (31) and of the vulva (2) were treated with high doses of Methotrexate. The objective response rate was 17% with a median duration of 3 months. Three toxic deaths occurred. The response rate of high dose methotrexate in advanced and prior treated cervix cancer does not differ from that reported in prior studies with conventional doses of intravenous methotrexate.


Electrocardiographic changes after adriamycin chemotherapy. The electrocardiograms of 146 patients with metastatic carcinoma of the breast were reviewed before, during, and after the patients received Adriamycin (Doxorubicin) chemotherapy (AD). The most significant electrocardiographic change occurred in the amplitude of the QRS voltage. Seven patients developed cardiomyopathy after AD and showed a significant decrease in QRS voltage. This decrease, however, was more severe at the onset of congestive heart failure that at conclusion of Adriamycin. In 35 patients with pleural effusion, there was an inverse relation between the extent of the effusion and the amplitude of QRS voltage in the absence of congestive heart failure. These results indicate that 1) the sudden and relatively severe decrease in QRS voltage with the onset of CHF limits the value of this ECG criterion for predicting early Adriamycin toxicity, and 2) caution should be exercised in the interpretation of QRS voltage changes in patients with significant pleural effusion.


Safety and efficacy of living BCG or BCG cell walls (CW) in the treatment of guinea pig hepatoma. Guinea pigs with established intradermal tumors and microscopic lymph node metastases were treated by intralesional injection of graded doses of living BCG or BCG CW. The lowest dose of living BCG used produced a significant cure rate and no grossly evident toxicity. An intermediate and the highest dose of living BCG used cured some animals but others lost weight and a significant number died as a result of the treatment. Histologic examination of animals with significant weight loss showed fatty degeneration of the liver, granulomatous hepatitis and histiocytic infiltration of the spleen. None of the doses of BCG CW used was toxic and they were at least as effective as living BCG in intralesional treatment. In some experiments in which treatment was delayed it was found that the extent of disease required to render treatment ineffective was about the same for living BCG and for BCG CW.


Cellular immunity in Hodgkin's disease.

Cellular immunity in Hodgkin's disease. Defective cell-mediated immunity (CMI) occurs early in the course of Hodgkin's disease (HD) and may persist even two years after successful treatment. This has been confirmed by in vivo and in vitro tests performed on 51 untreated and 52 treated patients of HD. The grading of skin reponse in vivo to dinitrochlorobenzene (DNCB) correlated very well with the in vitro leukocyte migration inhibition (LMI) response against phytohemagglutinin (PHA). An inhibitory influence of HD patients' sera was demonstrated by LMI tests in vitro. The response of peripheral leukocytes from HD patients in the LMI tests could be augmented in vitro by addition of levamisole (an immuno-potentiator) to the culture medium, thus pointing to an intrinsic defect in Lymphocytes. The data indicate that defect at multiple sites in the immune system is responsible for persistent anergy in HD.


Systemic complications of MER immunotherapy of cancer: pulmonary granulomatosis and rash. BCG immunotherapy often has severe complications in cancer patients despite lack of toxicity in the immunocompetent individual. MER, a cell wall fraction of BCG, has been reported to cause immunopotentiation similar to that of BCG without equivalent toxicity. Recently, animal models have been reported to develop MER complications, especially disseminated granuloma formation, like those of BCG. For the past several years, MER has been used as adjuvant immunotherapy for treatment of malignant tumors with minimal systemic toxicity reported. A patient with malignant melanoma was treated with intralesional MER at the site of local metastases. He developed military pulmonary granulomatosis and a severe cutaneous eruption in association with MER therapy. The toxicities of BCG and MER therapy were compared with the pathogenesis of granuloma formation reviewed. This patient's complications were consistent with a hypersensitivity reaction to MER. Pulmonary granulomatosis and rash must be added to the list of known MER toxicities.


Adenomatoid odontogenic tumor: ultrastructural demonstration of two cell types and amyloid. A typical adenomatoid odontogenic tumor removed from a 13-year-old female was studied by light and electron microscopy. The tumor was composed of two types of epithelial cells: Type I cells were cuboidal and occurred in nests or formed ductlike structures and Type II cells were smaller and spindle shaped. The formation of extracellular masses of amyloid was found in association with Type I epithelial cells, and amyloid formation was not observed in association with Type II cells. Results suggest that the lesion is of enamel organ origin, derived from cells of the inner enamel epithelium at the pre-ameloblastic stage, stellate reticulum and stratum intermedium. The origin of this amyloid material is unknown; however, it may be of enamel protein origin which, like amyloid, may have a beta-protein conformation.


A comparative study including ultrastructure of intramuscular myxoma and myxoid liposarcoma. The clinicopathological data of ten intramuscular myxomas (IMM) and three myxoid liposarcomas (MLS) are presented with emphasis on their ultrastructure. Electron microscopy of three of the IMM demonstrated the principal cell of the IMM to be similar to a fibroblast with prominent R.E.R., well-developed Golgi apparatus and cytoplasmic filaments. Intracytoplasmic lipid droplets were very rare. The matrix was composed of finely granular material, collagen fibers, fibrils and few capillaries. In contrast with the IMM, the cytoplasm of the MLS contained extensive lipid droplets. Capillaries were very abundant and in close proximity to the lipoblasts. The distinction between an IMM and MLS is usually clear-cut by light microscopy. Dissimilar ultrastructural features provide additional data to separate these two tumors. Ultrastructure confirmed Enzinger's impression that the cell of origin of the IMM is similar to a fibroblast with features that support the hypothesis that these cells are the source of production of the excessive amount of mucopolysaccharides in the matrix.


Mixed Brenner and adenomatoid tumor of the testis: an ultrastructural study and histogenetic considerations. A previously unreported association of Brenner and adenomatoid tumor found in the tunica vaginalis testis is presented. Many ultrastructural features found in mesothelial cells such as intercellular spaces, deeply indented nuclei, tonofilaments and tight desmosomes, were also shared by cells present in both neoplastic patterns. The previous histogenetic origins ascribed to testicular Brenner tumors are discussed and the evidence for their origin in the mesothelium considered.


Numbers of mast cells in the axillary lymph nodes of breast cancer patients. This retrospective study of surgical specimens from forty-three consecutive patients treated by modified radical mastectomy for carcinoma of the breast shows that those patients surviving for at least sixty months after operation have a significantly greater number of mast cells in their axillary lymph nodes than the nonsurvivors (p less than .001). When expressed as cells/mm2 the survivors are associated with nodal concentrations greater than eleven mast cells per square millimeter while the nonsurvivors haver fewer than eleven mast cells/mm2. The results suggest that the number of nodal mast cells may be a readily quantitated host response of prognostic value in carcinoma of the breast.


Atypical lipoma, atypical intramuscular lipoma, and well differentiated retroperitoneal liposarcoma: a reappraisal of 30 cases formerly classified as well differentiated liposarcoma. Thirty cases formerly diagnosed as Grade I lipogenic liposarcoma (well differentiated liposarcoma) were reviewed. The basic histologic pattern in all was that of adult fat modified by the presence of cells with enlarged, hyperchromatic nuclei; in most cases there was also a component of myxoid and/or fibrous tissue. The length of follow-up ranged from two to 30 years. Nine of the tumors were located in the subcutaneous layer. None of these recurred after excision, not even those which were simply "shelled out," and none metastasized. The term "atypical lipoma" is proposed for this group. Thirteen were located within or between muscles of the limbs, limb girdles, and head and neck. Nine of these recurred at least one, but there were no metastases and no deaths due to tumor. These were designated "atypical intramuscular lipoma." The remaining eight originated in the retroperitoneum. Although none of these patients developed metastases, five suffered inoperable recurrence and three died as a result of the neoplasm. It is suggested that the term "well differentiated retroperitoneal liposarcoma" be retained for cases of this type.


Tumor antigen of human cervical squamous cell carcinoma: correlation of circulating levels with disease progress. A double-antibody radioimmunoassay method was used for serial determinations of a tumor-antigen (TA-4) of cervical squamous cell carcinoma, and the correlation of serum antigen levels with the disease progess was investigated in 23 patients with cervical squamous cell carcinoma. Ten cases with widespread metastases received radiotherapy and/or chemotherapy. Nine of these cases who showed progression of the disease had a corresponding increase in serum antigen levels, while one case who had regression of the disease showed a corresponding decrease in serum antigen levels. Thirteen patients received radical surgery, and in all of these, high pretreatment antigen levels declined to undetectable levels 1 or 2 weeks after surgery. A panel of coded sera from the NCI-Mayo Clinic Serum Bank was also studied for evaluating the specificity of the assay. Thirteen of 25 patients (52%) with cervical squamous cell carcinoma showed positive serum antigen levels, while only one of 58 control cases (1.7%) showed false-positive result. These results suggest that serial TA-4 determinations may provide a useful method for evaluating regression or progression of the disease.


Localized fibrous "mesothelioma" of pleura (submesothelial fibroma): a clinicopathologic study of 18 cases. Eighteen cases of solid fibrous tumors of the visceral pleura are studied. The series includes neoplasms of various sizes removed during the last 17 years at The Mount Sinai Hospital. Most tumors showed consistent gross and light microscopic features, such as encapsulation, preservation of pedicle, origin from visceral pleura, predominance of spindle cells with more or less hyalinized stroma, little or no pleomorphism or mitotic activity and no infiltration of the underlying lung. No sex predominance or relation to environmental pollutants was noted. Peak incidence was in the 6th decade of life. Total excision proved curative in all cases with no recurrences or complications. The cell of origin of this tumor appears to be a noncommitted mesenchymal cell present in the areolar tissue subjacent to the mesothelial lining.


Stomal polypoid hypertrophic gastritis: a polypoid gastric lesion at gastroenterostomy site. Four cases of stomal polyps (the so-called gastritis cystica polyposa--GCP) at the gastroenterostomy site were studied with detailed description of their macroscopic and histologic features. Thirty-eight cases with reresection of the stomach including the site of previous gastroenterostomy were used for a comparative study. In 25 (66%) of the 38 cases localized mucosal hypertrophy of varying degrees was recognized near the site of anastomosis, the morphology corresponding well, though of the smaller scale, to that of the GCP both macroscopically and histologically. Macroscopic and histologic continuity was traced between the minimal protrusion of the stomal gastric mucosa and the stomal polyps. It is suggested that the GCP is an extreme occasion of stomal gastritis characterized by atrophy and pseudopyloric gland metaplasia of the fundic glands often with their submucosal invasion and hyperplasia of the surface and foveolar epithelia, these being probably caused by reflux of duodenal contents. It must be borne in mind that such lesions do exist at the site of gastrojejunostomy to be strictly differentiated from malignancies at the gastric stump.


Germ cell neoplasms arising in gonadoblastomas. Gonadoblastomas have a propensity to give rise to germ cell neoplasms. This study analyzes the clinicopathologic findings in six phenotypic females with features of 46,XY pure gonadal dysgenesis who developed germinomas and other germ cell tumors in gonadoblastomas. All stages in the evolution of germinoma from the germ cells of gonadoblastoma were observed, including in situ, incipient, microinvasive, and metastatic varieties. Admixtures with teratoma and endodermal sinus tumor occurred in two patients. Germ cell tumors which originate in gonadoblastomas appear to have the same clinical behavior and response to therapy as those that arise de novo in the ovary, testis or extragonadal sites. Although it is debatable whether gonadoblastomas are true neoplasms or blastomatoid dysgenetic malformations, their potential for giving rise to fully malignant germ cell neoplasms must be recognized.


Blood group isoantigen deletion in carcinoma in situ of the urinary bladder. Blood group isoantigens A and H (O) were measured by the Secific Red Cell Adherence (SRCA) Test in nine radical cystectomy specimens removed from patients with extensive carcinoma in situ of the urinary bladder. All bladders had areas of histologically normal epithelium and areas of epithelial atypia in addition to the carcinoma in situ. In eight cases, tissue-associated blood group isoantigens were deleted in areas showing either atypia or carcinoma in situ. Isoantigens were present in these areas in the ninth case. Blood group isoantigens were absent in approximately half of the sections of histologically normal epithelium. We propose that areas of epithelium which appear benign according to conventional histological criteria but in which the blood group isoantigens are absent may represent areas of low grade carcinoma in situ.


Retroperitoneal paraganglioma and systemic amyloidosis: a case report. The case of a 50-year-old woman with a retroperitoneal paraganglioma and nephrotic syndrome is presented. After the tumor was removed deposits of amyloid material were observed in the paraganglioma, kidneys, and liver. There was no thrombosis of the renal vein. The clinical picture (fever and nephrotic syndrome) was similar to that of patients with amyloidosis associated with neoplasms. The nephrotic syndrome disappeared 8 months after the operation. The patient has not had fever or nephrotic manifestations over the past 4 years and is asymptomatic at the present time. We believe this to be the first reported case of paraganglioma associated with amyloidosis. Nor have we found any references to clinical and biological remission of the nephrotic syndrome following surgical removal of the concomitant neoplasm.


Ovarian cancer: incidence and case-control study. The incidence of ovarian cancer in Rochester, Minnesota over the 40-year period 1935 through 1974 was determined; and risk factors for epithelial ovarian cancer occurring in Rochester from 1945 to 1974 were examined in 116 patients and 464 controls. Among the characteristics studied, only nulliparity was found to be a significant risk factor--relative risk 1.8. Other suspected risk factors--including hypertension, obesity, age at menopause, prior therapeutic pelvic radiation, and prior exposure to exogenous estrogen--were found not to differ significantly between patients and controls. The ovarian cancer patients were found to have a significantly lower frequency of prior hysterectomy and of unilateral oophorectomy than the control group. Thus out data show that hysterectomy, even when one or both ovaries are preserved, is associated with a lower risk of subsequent ovarian cancer.


Squamous cell carcinoma in chronic hydradenitis suppurativa: a case report. A case of squamous cell carcinoma arising in hydradenitis suppurativa of over 20 years duration is described. Only 6 other cases are reported in the literature. Multiple sites of origin of malignancy is suspected in this case. Wide-spread bone metastases resulted in severe hypercalcemia. Palliation was obtained for a short period following chemotherapy and Adriamycin and Bleomycin. Autopsy studies showed metastatic tumor in almost all the organs of the body.


Group counseling in adult patients with advanced cancer. The effect of a structured, interdisciplinary group counseling program was studied in 30 newly diagnosed adult patients with advanced cancer. These were compared to 30 patients who did not undergo group counseling. Group counseling resulted in a significant improvement in patient perception and self-concept. This structured educational and psychological support program provided a mutal support experience for newly diagnosed patients with advanced cancer.


A histopathologic study on the effects of radiofrequency thermotherapy on malignant tumors of the lung. Malignant cells are thermosensitive but this does not fully explain the wide-spread pathologic changes observed in solid cancers following local therapy with radiofrequency heating. The histopathology of human malignant tumors after radiofrequency thermotherapy reveals not only the expected degeneration of tumor cells, but significant changes in the tumor's stroma as well. Stromal blood capillary walls are massively infiltrated with round cells and undergo degenerative changes leading to necrosis and obliteration of the tumor's vascular supply. Extensive infiltration of the stroma with the host's round cells occurs. Following focal necrosis and rupture of the stroma, pools of lymphocytes establish direct contact with heat damaged malignant cells leading to further destruction of persisting tumor. The histologic signs of the generalized breakdown of lung tumors consequent to radiofrequency heat-therapy bear close resemblance to lesions in acutely rejecting organ allografts.


Patterns of urinary excretion of modified nucleosides. A recently developed high-performance liquid chromatography method permits quantitative measurement of low levels of modified nucleosides in urine. We report on the patterns of excretion of seven modified nucleosides by normal subjects and cancer patients. It was found that the excretion of these nucleosides expressed as a function of creatinine concentration was constant, not episodic nor related to diet. Thus, randomly collected samples of urine are satisfactory sources for measurement of nucleosides, and the level of nucleosides is of significance when related to creatinine excretion. The constancy of the excretion of the modified nucleosides in normal subjects is quite remarkable. It implies strict metabolic control of transfer RNA turnover. The values for the individual nucleoside/creatinine ratios were found to be significantly elevated in the urine of colon cancer patients.


Purification and mechanism of action of macromomycin. Macromomycin (MCR), a polypeptide antibiotic previously shown to have antitumor activity in experimental tumors, has been purified into an electrophoretically homogeneous component with an approximate molecular weight of 12,500. MCR has alanine as an NH2-terminal amino acid, 4 cysteine residues, and no arginine or methionine residues. With a fluorescence assay and agarose gel electrophoresis, MCR was shown to induce strand breaks in PM2 DNA in vitro. 2-Mercaptoethanol inhibited the DNA cleavage activity of MCR. When incubated with Novikoff hepatoma ascites cells in tissue culture, MCR caused Novikoff hepatoma ascites cell DNA degradation as observed by the slower sedimentation of DNA on alkaline sucrose density gradient centrifugation when compared to untreated cell DNA. DNA synthesis in Novikoff hepatoma ascites cells was inhibited by 80% after a two-hr treatment with MCR (0.03 microgram/ml). RNA and protein syntheses were inhibited by 25 and less than 10%, respectively, at this concentration of drug. At a concentration of MCR (1.0 microgram/ml), syntheses of DNA and RNA in Novikoff hepatoma ascites cells were totally inhibited. The results of this study suggest that MCR may inhibit tumor cell growth by causing DNA breakage with subsequent inhibition of DNA and other macromolecule syntheses.


Expression of cell-mediated immunity and blocking factor using a new line of ovarian cancer cells in vitro. A cell-mediated cytotoxicity test, quantitated by postlabeling with tritiated thymidine, was used to asses immune reactivity of cancer patients to the HeW cell line derived from serous cystadenocarcinoma of the ovary. Lymphocytes from 71.4% of serous and mucinous cystadenocarcinoma patients demonstrated a cytotoxic response towards the HeW cells, whereas no reactivity was observed towards target cells derived from nonovarian cancer. These observations indicated that the HeW cells express tumor-associated antigen. In some patients bearing similar tumors, cytotoxicity was blocked by ascitic fluid from other patients with cystadenocarcinoma. In addition, antigen obtained from the spent culture fluid of HeW cells exhibited blocking activity in a typical dose-response fashion, suggesting that blocking factor may be free tumor-associated antigen or an antigen-specific suppressor molecule. Thus, blocking of the lymphocytotoxic response of cystadenocarcinoma patients towards HeW cells may be utilized to monitor the isolation of ovarian carcinoma-associated antigen.


Promoting effect of saccharin and DL-tryptophan in urinary bladder carcinogenesis. The existence of at least two stages in bladder carcinogenesis was evaluated in male Fischer rats using N-[14-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) fed for six weeks at a level of 0.2% of the diet as the initiator. Sodium saccharin and DL-tryptophan were fed at levels of 5 and 2% of the diet, respectively, as possible promoting chemicals, and they were fed either immediately after FANFT administration or after six weeks of FANFT plus six weeks of control diet. All surviving rats were killed at the end of two years. Both chemicals significantly increased the incidence of bladder tumors following FANFT feeding compared to six  weeks of FANFT feeding followed by control diet, and the results were similar whether saccharin or tryptophan feeding was started immediately after FANFT feeding was concluded or after a six-week delay. Saccharin was considerably more potent as a promoting agent than was tryptophan, inducing higher incidences of bladder tumors and having a shorter latent period. Long-term administration of FANFT induced a 100% incidence of bladder cancer. Sequential epithelial changes were observed by scanning and transmission electron microscopy as well as by light microscopy. Pleomorphic microvilli were present on the superficial cells of all tumors examined and on the surface cells of hyperplastic bladder epithelium after six weeks of FANFT plus six weeks of saccharin, but not after six weeks of FANFT and six weeks of control diet. Rats fed only saccharin tryptophan, or control diet did not have bladder tumors or pleomorphic microvilli on bladder epithelium. These data suggest that saccharin and tryptophan might act as tumor-promoting agents during bladder carcinogenesis.


Cell growth recovery after treatments at various supraoptimal temperatures. The growth recovery kinetics of HeLa cells was investigated after treatments at intermediate (43-45 degrees) or high (49-55 degrees) supraoptimal temperatures for various periods of time (2 to 300 min) or after irradiations with nanosecond infrared CO2-laser pulses at energy densities equivalent to very high temperatures rises. After treatments at intermediate temperatures, single cells developed into colonies smaller than those obtained from nonheated control cells. Daily incorporations of pulsed [3H]thymidine in whole populations and microscopic observations of individual cell proliferation revealed a complex growth recovery for both survivors and dying cells. In contrast, colonies arising from cells treated at temperatures above 49 degrees do not differ in size from those developed from controls, and growth evaluations by [3H]thymidine and microscopy demonstrated that survivors resume normal proliferation immediately after treatments. The results which are supported by other studies on the effects of dose fractionation and metabolism status on cell survival are further discussed in relation to the "multistep" kinetic model for the cell response to hyperthermia.


Acute ultrastructural effects of the antitumor antibiotic carminomycin on nucleoli of rat tissues. Male Sprague-Dawley rats were treated with carminomycin i.v. in doses ranging from 1 to 40 mg/kg. Within 1 hr after the administration of carminomycin, 20 mg/kg, nucleoli of cardiac and skeletal muscle cells were segregated, while nucleoli of liver parenchyma cells were unaffected. Three and one-half hr after drug administration, cardiac muscle nucleoli reverted to normal ultrastructure. However, some skeletal muscle cell nucleoli were still segregated. Following treatment with carminomycin, 10 mg/kg, no significant ultrastructural changes were observed. These results demonstrate that at sufficiently high doses carminomycin induces ultrastructural lesions in nucleoli of both cardiac and muscle cells. The dose of carminomycin required to produce nucleolar segregation in cardiac and skeletal muscle is 6 times greater than the dose of Adriamycin (3.5 mg/kg) required to induce equivalent alterations.


Fluctuations in nucleoside uptake and binding of the inhibitor of nucleoside transport, nitrobenzylthioinosine, during the replication cycle of HeLa cells. Binding of the potent nucleoside transport inhibitor 6-[(4-nitrobenzyl)thio]-9-beta-D-ribofuranosylpurine (NBMPR) and rates of uptake of several nucleosides were examined at 4-hr intervals during the replication cycle of HeLa S3 cells. Monolayer cultures of synchronous cells, obtained by mitotic detachment, were assayed for high-affinity binding of NBMPR and for rates of uptake of thymidine, uridine, cytidine, adenosine, inosine, and guanosine. The number of NBMPR binding sites per cell doubled between 4 and 16 hr after detachment (late G1 and S phase); during this interval, V max 'S for uptake of cytidine and adenosine doubled, and for uridine and thymidine uptake increased about 4- and 8-fold, respectively. Rates of inosine and guanosine uptake at extracellular concentrations below saturation increased 2-fold between G1 and S phase of the cell cycle. Km 'S for cellular uptake of thymidine, uridine, cytidine, and adenosine did not change with progress through the cycle. The results presented suggest that changes in nucleoside uptake during the HeLa cell cycle were due, in part, to changes in the activity of NBMPR-sensitive transport elements in the membrane.


Tumor angiogenesis activity in clonal cells transformed by bovine adenovirus type 3. Four different sets of clonal cells transformed by bovine adenovirus type 3 and its oncogenic DNA fragments, and their clonal normal counterparts, were tested for tumor angiogenesis activity. The activity was assayed by measuring the host-mediated vascular response of a chorioallantoic membrane to the cell suspension separated from the vascular bed by a Millipore filter. Angiogenesis activity due to inflammation reaction was prevented by using corticosteroids. All of the transformed cells tested induced strong vascular responses as compared with their corresponding clonal normal cells. Cell dose and time dependency for the expression of the activity were also shown.


Differential effect of adriamycin on DNA replicative and repair synthesis in cultured neonatal rat cardiac cells. The effect of the potent antitumor antiobiotic Adriamycin (ADM) on DNA replication and unscheduled DNA synthesis in cultured rat cardiac cells was investigated. Autoradiography and [3H]thymidine incorporation studies were carried out on parallel cultures. DNA replication was depressed for up to 6 days following a 3-hr pulse of ADM administration. An ADM concentration of 1 microgram/ml which was effective in reducing replicative DNA synthesis by as much as 75% did not reduce the ability of cardiac cells to repair UV-damaged DNA. However, cells exposed to higher ADM concentrations failed to undergo significant UV-induced repair. In the absence of UV treatment, ADM did not stimulate unscheduled DNA synthesis. To account for the differential response of the cardiac cell cultures to replicate and repair DNA, we propose that ADM exerts a localized effect on DNA synthesis covering a region proximal to its primary intercalation site.


Comparison of mutagenicity, antitumor activity, and chemical properties of selected nitrosoureas and nitrosoamides. A number of nitrosoureas and nitrosoamides have been compared with respect to mutagenicity for Salmonella typhimurium, in vitro cytotoxicity, in vivo toxicity and antitumor activity against murine L1210 leukemia, and chemical properties. Despite chemical similarities between the nitrosoureas and nitrosoamides, they show important differences in biological activity. Some of the nitrosoureas are very active antitumor agents, and they are less mutagenic than are the corresponding nitrosoamides, which lack antitumor activity.


Effects of cis-dichlorodiammineplatinum(II) on human colon carcinoma cells in vitro. The lethal effects of cis-dichlorodiammineplatinum(II) were investigated on an established human colon carcinoma cell line. cis-Dichlorodiammineplatinum(II) was one of the most efficient antineoplastic agents tested thus far on this human colon carcinoma cell line. Survival of exponentially growing cells exposed to increasing concentrations of the drug (both in medium or in Hanks' balanced salt solution) was of the threshold exponential type (Dq = 1.2 microgram/ml, 1 hr; Do = 3.5 microgram/ml, 1 hr). Stationary-phase cells were extremely sensitive to the drug, and the survival curve demonstrated a simple exponential pattern (Do = 3.9 microgram/ml, 1 hr). Long-term exposure to low concentrations of cis-dichlorodiammineplatinum induced a high degree of killing, with only 0.5% of the cells surviving after incubation for 24 hr with 2 microgram/ml. Cells were unable to recover from potentially lethal or sublethal damages induced by the drug.


N-(4-Hydroxyphenyl)retinamide, a new retinoid for prevention of breast cancer in the rat. The synethesis of a new retinoid, N-(4-hydroxyphenyl)-all-trans-retinamide, which has useful biological properties, is described. This retinoid was more potent than retinyl acetate in reversing keratinization caused by retinoid deficiency in tracheal organ culture. It was markedly less toxic than retinyl acetate when fed p.o. to rats over 2-week or 6-month periods. It was an effective agent for inhibition of the development of breast cancer induced in rats by N-nitroso-N-methylurea, although it was not as potent as retinyl acetate in this regard. However, the lesser toxicity of 4-hydroxyphenylretinamide makes it a superior agent for prevention of breast cancer. High-pressure liquid chromatographic analyses of liver and breast extracts from rats treated for 6 months with retinoids show the pharmacokinetic basis for the superiority of 4-hydroxyphenylretinamide; this retinoid and its metabolites were found in high concentrations in breast tissue, without any measurable accumulation in the liver or evident liver toxicity. In contrast, chronic feeding of retinyl acetate caused marked deposition of retinyl esters in the liver and severe hepatotoxicity. Whole mounts of rat mammary glands, made after chronic feeding of 4-hydroxyphenylretinamide, showed that it had a marked antiproliferative effect on mammary epithelium.


Physical and chemical composition of subcellular fractions from tumor cells treated with metabolic inhibitors or hormones. Line 10 guinea pig hepatoma cells are resistant to killing by antibody plus guinea pig complement but not by antibody plus human complement. Agents that increase (metabolic inhibitors) or decrease (hormones) the sensitivity of the cells to killing by antibody plus complement were examined for their effects on the chemical, physical, and enzymic composition of the cells. The effects of these agents on the chemical and enzymic characteristics of isolated plasma membrane and intracellular fractions of these cells were also measured. Adriamycin treatment resulted in a decrease in the amount of ribonucleoprotein and smooth endoplasmic reticulum isolated from the cells as compared to untreated cells. Hormone (insulin or hydrocortisone)-treated cells were enhanced in their yield of this fraction but were decreased in their yield of mitochondria as compared to controls. Generally, the drug-treated cells were decreased, whereas hormone-treated cells were enhanced, in protein, lipid phosphate, and total phosphate content, as compared to untreated controls. This pattern was also noted in the plasma membrane fraction of the cells and, in several cases, in intracellular membrane fractions. These data suggest that the protein, lipid phosphate, and total phosphate content of the plasma membrane and intracellular membranes of these cells may correlate with their ability to resist humoral immune attack.


Formation and removal of specific acetylaminofluorene-DNA adducts in mouse and human cells measured by radioimmunoassay. Rabbit antiserum prepared against N-(guanosin-8-yl)-acetylaminofluorene was utilized in radioimmunoassay to detect formation and removal of C-8 adducts from the DNA of cultured cells exposed to N-acetoxy-2-acetylaminoflorene. The assay was able to quantitate both acetylated and deacetylated C-8 adducts between 0.5 and 5 pmol while the N2 adduct, 3-(deoxyguanosin-N2-yl)acetylaminofluorene, was not detected below 160 pmol. By varying the proportions of acetylated and deacetylated C-8 adducts in the radioimmunoassay, a series of standard curves were developed from which the relative proportion of each adduct could be determined in unknown mixtures. DNA from mouse epidermal cells and human skin fibroblasts exposed to N-acetoxy-2-acetylaminofluorene in culture contained only 3 and 5% respectively, of the C-8 adduct in the acetylated form. Quantitation by radioimmunoassay of total C-8 adducts bound to DNA yielded values approximately 25% lower than total carcinogen binding determined by radiolabeling. When removal of C-8 adducts was followed over a 23-hr, carcinogen-free culture period, mouse and human cells removed 40 and 50%, respectively, of bound acetylated and deacetylated C-8 adducts. These studies demonstrate the versatility of radioimmunoassay as a molecular probe for studies of chemical carcinogens.


Spermine-induced variations in the adenosine 5'-diphosphate ribosylation patterns of nuclear proteins from rat liver and hepatoma. Rat liver and hepatoma nuclei were incubated in vitro with [3H]nicotinamide adenine dinucleotide to allow synthesis of a polymer of adenosine diphosphoribose subunits joined in an 1',2' ribose-ribose linkage. The addition of 1 mM spermine altered the adenosine 5'-diphosphate (ADP) ribosylation patterns of nuclear proteins in hepatoma, host liver, and regenerating liver. Spermine-treated nuclei showed a greater incorporation of ADP-ribose into H1 histones and nonhistone nuclear proteins with isoelectric points between pH 3.0 and 6.0 when separated on polyacrylamide gels. Conversely, a large reduction in ADP ribosylation was seen in core histones (H2A, H2B, and H3) from the same nuclei. The proportion of ADP-ribose incorporated into histones was reduced in the nuclei from proliferating cells relative to their respective control livers. These results imply that polyamines, which are higher in concentration in rapidly dividing cells, may elicit a regulatory function by causing the preferential ADP ribosylation of H1 histones, as well as the more acidic of the nuclear proteins.


Evidence for a new mechanism of cytotoxicity of 1-beta-D arabinofuranosylcytosine. Inhibition of DNA synthesis by a pulse of 1-beta-D-arabinofuranosylcytosine (ara-C) results in reinitiation of DNA replication in DNA segments replicated earlier in that S phase and hence double replication of some DNA segments. Experiments were with tissue culture cell lines of human origin. DNA replicated early in S phase was labeled with a pulse of [3H]deoxycytidine with the cells pulsed later in S phase with ara-C. The DNA replicated after the time of the ara-C pulse was density labeled with 5-bromodeoxyuridine. Reinitiation of DNA replication in the already replicated [3H]DNA segments was demonstrated using CsCl density gradient analysis by an increase in the 3H label present in the light-heavy peak of semiconservatively replicated DNA. Also, in DNA of control cells, all of the 3H was in the same strand of the light-heavy DNA duplex as was the 5-bromodeoxyuridine, as shown by alkaline CsCl density gradient analysis of purified light-heavy DNA. However, after a pulse of ara-C, utilization of [3H]DNA strands as template strands was demonstrated by the presence of 3H label at the density of unsubstituted DNA chains in alkaline CsCl gradients of the purified light-heavy DNA. This double replication phenomenon can explain certain chromosomal abnormalities induced by ara-C.


Recovery of 2'-deoxycoformycin-inhibited adenosine deaminase of mouse erythrocytes and leukemia L1210 in vivo. The antibiotic 2'-deoxycoformycin, a potent inhibitor of adenosine deaminase, has potential as a chemotherapeutic agent. Injection of 2'-deoxycoformycin i.v. (0.2 mg/kg) to mice bearing ascites L1210 leukemia cells completely inhibits adenosine deaminase in both erythrocytes and L1210 cells. The recovery of the enzymic activity is markedly different in the two tissues. The recovery is very slow in erythrocytes (13% in 48 hr), whereas 80% recovery occurs during the same time interval in L1210 cells. This marked difference in the recovery of the enzyme in different tissues may play a role in the pharmacological and chemotherapeutic behavior of this drug.


Renal excretion kinetics of high-dose cis-dichlorodiammineplatinum(II) administered with hydration and mannitol diuresis. The effect of hydration and mannitol diuresis on the nephrotoxicity and renal excretion kinetics of high-dose cis-dichlorodiammineplatinum(II) was examined in seven men undergoing treatment for testicular or bladder carcinoma. The elimination half-life averaged 26.8 hours, shorter than values previously reported from studies not employing hydration and diuresis. In addition, no signs of nephrotoxicity were observed. The results suggest that hydration and mannitol diuresis decrease the nephrotoxicity of cis-dichlorodiammineplatinum(II) by increasing its rate of elimination.


Pharmacology and toxicology of alpha-2'-deoxythioguanosine in cancer patients. alpha-2'-Deoxythioguanosine (alpha-TGdR) was administered as a single dose to 13 cancer patients in 18 experiments at dose levels of 150--1500 mg/m2 and as a daily dose to 22 patients in 42 experiments at dose levels of 100--4000 mg/m2/day X 5 days. No significant toxicity was observed. Blood levels and rates of excretion were determined with radiosulfur-labeled alpha-TGdR. Approximately 80% of the dose was excreted in the urine in 24 hours, initially as unchanged alpha-TGdR and increasingly as metabolites. Metabolites appear to be nucleosides and do not include 6-thioguanine, 6-thioxanthine, or 6-thiouric acid to any measurable extent. Small amounts of the alpha-TGdR in blood samples were bound to albumin and to erythrocyte membranes. Blood plasma levels of alpha-TGdR at the highest doses were in the range of 200--300 micrometer, declining in 24 hours to 67--124 micrometer.


Enzymatic assay for the antitumor agent-N-(phosphonacetyl)-L-aspartic acid (PALA). A method is described for the assay of N-(phosphonacetyl)-L-aspartic acid in plasma or urine, based on inhibition of partially purified aspartate carbamoyltransferase from rat liver. Concentrations of N-(phosphonacetyl)-L-aspartic acid as low as 0.1 microgram/ml can be detected. Plasma disappearance and urinary excretion curves from one patient are shown as examples of the application of the method.


BISACK analysis of the phytohaemagglutinin-induced proliferation of human peripheral lymphocytes. The rate of stimulation as well as subsequent cell cycle duration was examined in phytohaemagglutinin-stimulated human peripheral lymphocytes grown in vitro in the presence of non-inhibotory concentrations of bromodeoxyuridine. After incorporation of this heavy atom analogue of thymidine into replicating cellular DNA, it was possible to identify unequivocally metaphase cells which had replicated for one, two and three or more cells cycles. Utlizing this technique, distribution curves were obtained for the appearance of metaphase cells in successive generations, were analysed by a computer simulation model, and the rate of stimulation (4.5% per hr of the reminaing unstimulated population) and cell cycle duration (12.3 hr) were determined. The results were compared with those obtained by autoradiography and the possible relationship to the 'transition probability' model for cellular proliferation is discussed.


Tumour cell proliferation in relation to the vasculature. The proliferation pattern of a transplantable mouse mammary carcinoma has been studied in relation to its macroscopic and microscopic structure. No significant differences were seen in the labelling or mitotic indices or in the percentage labelled mitoses curves for the peripheral 2.0 mm rim or for the central tumour core. When these parameters were scored for cells classified according to their position in relation to capillaries or to necrotic regions, marked differences were observed in all the parameters. Higher labelling and mitotic indices and higher grain counts were seen adjacent to the capillaries. These appear to result from a shorter cell cycle duration and a higher growth fraction. The variation in cell cycle is mainly due to a change in the duration of G1.


Growth of mouse and human bone marrow in diffusion chambers in mice. Development of myeloid and erythroid colonies and proliferation of myeloid stem cells in cyclophosphamide- and erythropoietin-treated mice. Both murine and human bone marrow cells were cultured in plasma clots which were formed inside diffusion chambers implanted into cyclophosphamide- and saline-treated mice. After an initial fall, the number of mouse bone marrow cells and numbers of mouse myeloid stem cells (CFU-C) and agar cluster-forming units rose faster in the cyclophosphamide-treated animals. These hosts also favored formation of myeloid (CFU-D-G) and erythroid (CFR-D-E) colonies and myeloid higher than those of CFU-C from the same marrow population. These observations suggest the existence of humoral factors stimulating granulocyte progenitor cell replication and differentiation. At its best the increment of CFU-D-E number was equivalent to that caused by a single 0.1 unit erythropoietin dose. Culture of normal human marrow cells resulted in colonies in the plasma clot containing only granulocytes and macrophages. Cyclophosphamide-treated host animals were essential for human CFU-D-G development. Plating efficiency for human marrow myeloid colonies was better in the conventional in vitro agar cultures than in diffusion chambers.


Morphological, histochemical and biochemical studies on germ cell mitochondria of normal rats. Morphological changes in rat germ cell mitochondria are described. In diplotene and secondary spermatocytes and in the spermatids of the Golgi, cap and acrosomal phases, the mitochondria take on a rounded appearance with the inner space containing the matrix flattened against the outer membrane and the intracristal spaces considerably swollen ("condensed" mitochondria). Functional studies on "condensed" mitochondria isolated from the germ cells of normal rats have been performed. The following parameters have been evaluated: ADP/O ratio, respiratory control ratio (RCR) and ADP affinity. The ADP/O values found in the presence of various substrates are in agreement with the theoretical figures. The RCR is remarkably high. Moreover, the ADP affinity of these mitochondria is very high, as demonstrated by the low values of the "apparent Km". These biochemical findings, which demonstrate a high oxidative capacity coupled with a marked phosphorylation, suggest that the "condensed" appearance of germ cell mitochondria is the expression of an active functional state.


Immunocytochemical identification of vasopressinergic and oxytocinergic neurons in the hypothalamus of the cat. Our immunocytochemical investigation of the magnocellular neuroendocrine cells in the cat hypothalamus reveals a mixture of vasopressin (VP)- and oxytocin (OT)-containing neurons in the supraoptic (NSO), the paraventricular (NPV) and in five accessory nuclei (NAC). We describe the lateral hypothalamic nucleus (NLH), a new accessory nucleus, lying at the junction of the internal capsule and pallidum, and possibly involved in drinking behavior. Previously characterized incompletely in mammals, the four other accessory nuclei consist of the circularis (NC), anterior fornical (NAF), posterior fornical (NPF) and retrochiasmatic (NRC). The two peptidergic cell types, VP and OT, are equally mixed in the NPV and the NAC, but in the NSO VP neurons predominate. The perikarya of these VP and OT neurons do not show distinct morphological differences at the level of light microscopy. The organization of magnocellular neuroscretory neurons in the cat hypothalamus closely resembles that described in other mammals with the exception of the unique presence of the lateral hypothalamic accessory nucleus.


Metals and phosphate in the chloragosomes of Lumbricus terrestris and their possible physiological significance. The chloragog cells of the earthworm Lumbricus terrestris contain numerous granules, chloragogomes, which were analyzed for metals and phosphate by histochemistry, by use of an electron microscope X-ray microprobe (EMMA) and by chemical analysis of chloragosome preparations. Inorganic and organic phosphate each accounts for about 3% of the chloragosome dry mass, Ca for 2--3%, Zn for 1--3% and Mg for 0.2--0.4%. Carbonate is not present in chloragosomes. The average molar ratios Ca:Mg:Zn:total PO4 are 1:0.1:0.3:1. The Ca:PO4 ratio is fairly constant (correlation coefficient 0.99), while the Zn:PO4 ratio varies considerably. It is concluded that Ca is bound in the form of inorganic CaHPO4 and organic R-OPO3Ca (or possibly Ca-polyphosphate complexes). Mg may also be phosphate-bound, while Zn probably is not. Chemical analysis of the calciferous glands revealed a high concentration of Ca, small amounts of Mg and phosphate, but no Zn. It is concluded that Zn is not excreted through the calciferous gland. Storage of Ca in the chloragocytes and excretion of CaCO3 by the calciferous gland may be physiologically linked. Regulation of the concentrations of Ca and HCO3- ions in the blood and coelomic fluid may assist in equalization of osmotic pressures during dehydration and rehydration. This regulation may be a major function of the chloragosomes. The chloragosomes were discussed in relation to the "spherites" of various arthropods and molluscs and to the "cytosomes" of anoxia-tolerating molluscs.


Metabolism of glycoconjugates in hypothalamic neurons and glial cells: comparison of incorporation of [3H]fucose and [3H]N-acetylmannosamine by electron microscopic autoradiography. Incorporation of two glycoconjugate precursors, [3H]fucose and [3H]N-acetylmannosamine, in the arcuate nucleus of the rat hypothalamus is compared 30 min after intraventricular administration. Electron microscopic autoradiographs were analyzed by a sampling technique which provides information about (i) the distribution of radioactivity in the tissue, (ii) the relative volumes, and (iii) the amount of radioactivity per unit volume of the various tissue compartments. With both precursors, the highest incorporation was found in glial cell bodies, being about five to six times that of neurons. This holds true for all three types of glia. The concentration of radioactivity in the neuropil consisting of neuronal and glial processes was exceedingly low, although, due to its large volume, it contained the highest fraction of total tissue radioactivity. The limitations imposed on the interpretation of the data in terms of synthesis of fucosyl- and sialoglycoconjugates are discussed. It is proposed that the observations support the concept that high rates of synthesis for glycoproteins (and possibly gangliosides) are a characteristic metabolic property of glial cells.


Effects of heat acclimation on the secretory products of the submaxillary gland of the rat. An ultrastructural and biochemical study. Secretory granule area and glycoprotein concentration of the saliva in the submaxillary gland of rats were measured during various stages of acclimation to heat at 34 +/- 1 degrees C. Granule size decreased by 18% during the first five days of heat acclimation (0.025 less than p less than 0.05) after which period it increased to reach 118% of the control levels after 28 days (p less than 0.05). Glycoprotein concentration in the saliva of stimulated glands rose above control levels, reaching a maximum between the 2nd and 5th day of acclimation (p less than 0.05). It was concluded that the initial decrease in granule size reflects a decrease in glycoprotein content following an increase in salivary flow known to occur at high ambient temperatures. The subsequent increase in granule size is considered an adaptation of the gland to continuous stimulation. The rise in salivary glycoprotein concentration suggests increased efficiency of the secretory mechanism.


Scanning electron microscopy of microcorrosion casts of the vascular bed in the skin of the spotted salamander, Salamandra salamandra L. Microcorrosion casts of blood vessels in the skin of the spotted salamander, Salamandra salamandra L., were studied using scanning eletron microscopy. The investigated vessels include a subepidermal network of respiratory capillaries and the vessels of poison glands. A hypothesis is proposed, according to which both types of vascular beds possess a common origin. Probable factors involved in the differentiation of the primary netword of the subepidermal vessels in larvae are indicated and speculation concerning the supposed mosaic type of circulation in the capillaries of the subepidermal respiratory bed is presented.


A system of intraependymal cisternae along the margins of the median eminence in the rat: structure, three-dimensional arrangement and ontogeny. Structure, three-dimensional arrangement and ontogeny of large intracellular cisternae located in the median eminence region of the rat hypothalamus were studied using toluidin-blue stained semithin sections and electron microscopy. The cisternae occur along the projections of ependymal cells lining the ventral portion of the third ventricle (infundibular recess). Small cisternae can be seen close to the ventricle, whereas larger ones, divided into smaller compartments by thin septa, cluster near the surface of the hypothalamus. The cisternae are encompassed by a thin layer of cytoplasm to which axon terminals containing synaptic and dense core vesicles are closely related. Cisternae are arranged around the median eminence in a characteristic pattern. They occupy the midline in the retrochiasmatic area, flank both margins of the median eminence and extend caudally behind the origin of the pituitary stalk. The cisternae appear first between the 15th and 17th postnatal days. At about the 30th day their size and distribution resemble the situation observed in adult animals. The ependymal cisternae are suggested to be closely related to the luteinizing-hormone releasing-hormone (lH-RH)-containing fibers.


The midgut of Tomocerus minor Lubbock (insecta, collembola): ultrastructure, cytochemistry, ageing and renewal during a moulting cycle. The midgut cells of Tomocerus minor (Insecta, Collembola) were examined with the electron microscope and cytochemically. The midgut epithelium consists of a series of cells characterised by numerous mineral concretions scattered throughout the cytoplasm. Mitochondria are abundant; microvilli are well developed at the apical surface of the cell. A zonula continua (continuous junction) characterises the apical contact region of these cells. Polysaccharides, glycoproteins and carbohydrate components have been demonstrated on the surface of microvilli. Peritrophic membranes surround the food bolus and preserve midgut cells from mechanical abrasion. Lysosomes are present during the alimentary period and show strong acid phosphatase activity. During an intermoulting cycle, two stages can be observed: (1) the postexuvial feeding period during which cytoplasmic extrusions appear at the apical part of the cell: lysosomes increase in number and autophagic vacuoles appear. (2) The preexuvial fasting period; a new epithelium grows beneath the old one and pushes it into the lumen. Degeneration processes can be observed in the old epithelium. This excretory reactivity of the midgut epithelium has been compared to the cycle of the cuticle.


Three dimensional organization of mammalian adrenal cortex. A scanning electron microscopic study. The adrenal cortex of different mammals was studied by SEM in order to demonstrate its actual three-dimensional organization. In the rat, as well as in the cat and pig, the adrenal cortex appeared as a "tunnelled continuum" of polyhedral cells arranged in plate-like structures (laminae). This laminar arrangement was more evident in the inner fasciculate and reticular zones where the cortex revealed a striking similarity to liver tissue. The polyhedral cells of all cortical zones possessed regular facets populated by small pits, larger invaginations and numerous microvilli with the exception of very short and smooth areas probably corresponding to attachment zones and/or gap junctions. This cellular architecture produced a labyrinthic system of intercellular channels or lacunae in which the capillaries were suspended. The pericapillary areas of this labyrinth contained microvilli, amorphous material, a delicate net of fibrils and occasional cells. The intercellular compartment of this lacunar system was mainly bordered by numerous microvilli arising from endocrine cells. The luminal surface of the capillary wall showed not only irregularly protruding margins (interpretable as endothelial junctions) but also clearly overlapping and flattened endothelial extensions. In all the animals and areas of the adrenal cortex examined, the endothelial wall was provided with abundant clusters of small fenestrations (about 50 nm in diameter) generally arranged in sieve plates. Larger fenestrations were noted mainly in the fasciculate and reticular zones of the cat and pig and occasionally in the rat. A final point related to the nature and significance of sinusoidal fenestrations was the occurrence of irregularly shaped and intracapillary located cells mainly noted in the deeper zones of the fasciculate and reticular zones of the gland. These elements--possessing the surface characteristics of macrophages--were observed, with their irregular and slender evaginations, in close proximity to the large fenestrations in a manner reminiscent of Kupffer cells within the lumen of liver sinusoids.


Electron microscopic investigation of the formation of spermatophores of Armadillidium vulgare. An ultrastructural investigation of the formation and morphology of spermatophores of the isopod, Armadillidium vulgare, has been conducted. Armadillidium spermatozoa are organized into bundles ensheathed by a specialized structure that functions as a spermatophore. The spermatophore is an extracellular investment consisting of two components: (1) a cone-shaped assembly of longitudinally aligned, 400 to 450 A diameter extracellular tubules, extending from the area rostral to sperm acrosomes to the region of sperm nuclei; and (2) matrix material, which surrounds spermatozoa for the entire length of the bundle. Morphological evidence suggests the participation of the rough endoplasmic reticulum and Golgi apparatus of testicular follicle cells in the production of tubular and matrix components of the spermatophore. Although the organization of spermatophores is similar throughout the male reproductive tract, the morphology of the matrix material appears to change at lower regions of the tract.


Ultrastructural and biochemical characteristics of Leydig cells from newborn androgen-insensitive rats. Leydig cells of testis of newborn pseudohermaphrodite (tfm) rats have an ultrastructure similar to that of the normal, containing well developed organelles and inclusions. The cytoplasm is filled with smooth endoplasmic reticulum forming a network of interconnected tubules. Lipid droplets are surrounded by cisternae of smooth endoplasmic reticulum and are in close association with pleomorphic mitochondria. Many of the latter are cup-shaped and have tubular cristae and intramitochondrial dense bodies. Essentially, these are characteristics of normal Leydig cells. Accordingly, the production of testosterone by testes from newborn tfm rats is the same as that by testes from normal newborns and adults. However, it is significantly higher than that by testes of tfm adults. Also, the plasma testosterone levels of newborn tfm rats are the same as in the normal newborn, but lower than in normal adults and much lower than in adult tfm animals. Thus, since in the tfm rat the morphology of Leydig cells, androgen production, and maintenance of plasma levels of testosterone are normal in the newborn, but become abnormal with advancing age, it appears that defective androgen action rather than insufficient androgen production is the cause of male pseudohermaphroditism.


Resorption of uncalcified cartilage in the diaphysis of the chick embryo tibia. The resorption of the uncalcified cartilage matrix of the middle third of the diaphysis in the chick embryo tibia has been studied using histological, histochemical and electron microscopic techniques. The first stage in the resorption process affects the periosteal bone, which is breached by osteoclasts at one or several points. Capillary vessels and clear, apparently undifferentiated cells penetrate through the holes so formed and reach the cartilage. The loss of acid proteoglycans to a depth of 10--20 micrometer into the matrix is the first sign of cartilage resorption; it is followed by the digestion of collagen fibrils, the opening of cell lacunae, chondrocyte degeneration and fragmentation and, lastly, the complete dissolution of the cartilage. This process is mediated by cells which probably derive from  perivascular elements. Most of these cells have an undifferentiated appearance, but they have macrophagic properties, as is shown by phagocytotic activity along their plasma membrane, by the presence of lysosome-like bodies in their cytoplasm, and by their intense acid phosphatase activity. Resorption by giant cells of chondroclastic type only occurs at a late stage.


Identification of G6PDH-active sinusoidal cells as Kupffer cells in the rat liver. The aim of this study was to identify the G6PDH-active sinusoidal cells in the rat liver described by Rieder et al. (1978). Because of their number and distribution in the liver parenchyma, endothelial cells and pit cells could be excluded. Fat-storing cells were specifically marked by vital staining with vitamin A and identified by fluorescence microscopy. Kupffer cells could be detected after vital staining with carmine. Both staining methods allowed a subsequent incubation for the demonstration of G6PDH activity in the same unfixed cryostat section. Whereas more than 80% of the fluorescent particles were found outside the enzyme-positive cells, all G6PDH-active cells contained carmine particles. After counting the G6PDH-active cells, an estimation of 0.217 x 10(8) cells/g liver tissue was obtained. The results indicate that high G6PDH activity is common to all Kupffer cells, and is therefore a highly specific marker enzyme for this class of sinusoidal liver cells.


New observations on the fine structure of the liver in goldfish (Carassius auratus). A re-examination of goldfish liver was made through the use of SEM of fractured samples and TEM of ultrathin-sections and freeze-etch replicas. Several new hepatic fine structures described in the present study are morphologically similar to those reported previously in many higher vertebrates including mammals. Hepatic sinusoids of goldfish contain fenestrations which are arranged into sieve plates. Although the hepatic plates are made up of two layers of hepatocytes, the parenchymal cells of gold fish liver are morphologically similar to mammalian hepatocytes, particularly with respect to the sinusoidal surfaces which are studded with numerous microvilli. The intercellular surfaces of hepatocytes have both nexus and demosomal junctions, similar to those found in various epithelial cells of higher vertebrates, between the openings of the intracellular bile canaliculi and the intralobular bile ductules which are situated in the center of the bicellular hepatic plate.


Ultrastructural and histochemical investigation of the terminal capillaries in the spleen of the carp (Cyprinus carpio L.). In the spleen of the carp arterial capillaries of a highly differentiated structure have been studied by light and electron microscopy. These capillaries share various structural characteristics with the sheathed capillaries (ellipsoids of Schweigger-Seidel) of higher vertebrates. The long arterial capillaries of the carp spleen are provided with cuboidal endothelial cells containing filaments approximately 7 nm in diameter. There is no basal lamina. The endothelial cells form various types of cell junctions, but there are also extensive areas without any junctions. Here, a free passage is possible between the capillary lumen and the subendothelial space. The capillaries possess a single-layered sheath of macrophages. Characteristically, the sheath macrophages posses long and slender cell processes forming a loose framework, the meshes of which are filled with lymphocytes and spindle cells. The sheath macrophages show a zone of ectoplasm rich in filaments. The also contain numerous phagolysosomes rich in hydrolytic enzymes, as identified histochemically. The sheath is sharply limited against the pulp by a thick layer of collagen fibers.


A comparative study on the innervation and the vascularization of the bulbus arteriosus in teleost fish. The structure of the bulbus arteriosus of a wide range of teleost fish is described with particular reference to the vascularization and innervation. The adventitia of the organ consists of blood vessels and large nerve bundles in a collagen matrix. The nerve bundles contain monoamines, and fluorescence studies show small terminal bundles penetrating the muscular media; this is confirmed by electron microscopy. The media consists of an extensive elastic tissue matrix with a spiral arrangement of smooth muscle cells joined end to end by desmosomes and presumed electronic junctions. The muscles cells are innervated only at the adventitia/media boundary and the significance of this innervation is discussed. It is proposed that there is a correlation between the degree of vascularization and innervation and the activity of a particular species of fish.


Stereologic studies on mitochondrial configuration in different organs of the rat. Mitochondria from different organs of the rat with configurations ultrastructurally resembling those of isolated mitochondria of known respiratory states have been subjected to stereologic analysis. Mitochondria were examined from mossy fibers of the granular layer of the cerebellar cortex (condensed state), of the pericentral hepatocytes (orthodox state), and of heart muscle and parietal cells of the gastric fundus (transitional state). In order to study the relationship between mitochondrial compartments and the internal membrane a "partition coefficient" was introduced, which expresses the volume of the matrix (E mm) and external compartment (E ocm) respectively per unit surface area of internal mitochondrial membrane. The sterologic parameters investigated, i.e. surface density of the mitochondrial membranes, volume density of the mitochondrial compartments and membranes, and partition coefficients generally agreed with the visual evaluation of mitochondrial ultrastructure. However, analysis of the coefficient of variation delta/x x 100% for E ocm and E mm has shown significantly greater variability in the mitochondria of the myocardium than in the gastric mitochondria, despite similar ultrastructure. It is suggested that stereologic methods, like time-lapse cinematography, give a compound picture of configurational variation and of its plasticity.


Molecular changes in the membranes of mouse erythroid cells accompanying differentiation. The development of the mouse erythroblast to a mature erythrocyte is accompanied by changes in the composition and properties of the plasma membranes of these cells. Using double fluorescence techniques, we have simultaneously determined the distribution of lectin receptors and spectrin on the membranes of these cells. The lateral mobility of the lectin receptors in the membranes decreases as differentiation proceeds, and this is accompanied by an increasing concentration of spectrin associated with the membranes. The most significant concentration of spectrin occurs, however, during the enucleation of the late erythroblast, where we observe a complete segregation of the spectrin to the incipient reticulocyte, as well as a previously observed enrichment of receptors for concanavalin A into the plasma membrane surrounding the extruding nucleus. On the basis of these and other observations, we explore the possible molecular mechanisms involved in erythroblast enucleation and the role of spectrin in the regulation of protein mobility in erythroid cell membranes.


The ovalbumin gene: alleles created by mutations in the intervening sequences of the natural gene. Two allelic forms of the natural chicken ovalbumin gene have been independently cloned. These alleles differ from each other by an Eco RI restriction cleavage site in one of the seven intervening sequences within the natural ovalbumin gene. Restriction endonuclease mapping and sequence analyses of these cloned genotypic alleles have shown identical sequence organization and molecular structures of the interspersed structural and intervening sequences except for the particular Eco RI cleavage site. Sequencing data of the cloned DNA suggest that this Eco RI site may be created or eliminated by a single base mutation in the intervening sequence of the ovalbumin gene. The occurrence of apparent homozygous and heterozygous allelic forms of the ovalbumin gene in individual hens and roosters within the same breed has been observed. 10 and 40% of the chickens examined are homozygous for the ovalbumin gene with and without the extra Eco RI site, respectively, while 50% of them are heterozygous. Further analysis of individual chicken DNA cleaved by restriction endonuclease Hae III has revealed that there may be a series of such mutational variations within the ovalbumin gene. We have identified two Hae III cleavage sites that do not occur in all of the chickens, thus giving rise to several additional allelic variations of the ovalbumin gene. At least one of these Hae III sites is situated in the intervening sequence of the ovalbumin gene, and its lcoation has been mapped. Such allelic variations must be taken into consideration when determining eucaryotic gene structure by restriction mapping of the genomic DNA. Furthermore, this type of mutation within the intervening sequences of an eucaryotic gene has no known phenotypic manifestation. It represents an extrastructural silent mutation that must be taken account of in studies to estimate the rates of eucaryotic gene sequence divergence during evolution.


A murine leukemia virus mutant with a temperature-sensitive defect in membrane glycoprotein synthesis. Cells infected with a temperature-sensitive mutant (ts-26) of Rauscher murine leukemia virus (R-MuLV) or with wild-type virus were labeled with 35S-methionine, and cell extracts were examined for radioactive polypeptides which could be precipitated by monospecific antisera to viral proteins. When shifted from permissive (31 degrees C) to nonpermissive (39 degrees C) temperature, cells infected with ts-26 rapidly begin to accumulate gPr90enr, the glycoprotein precursor to the membrane envelope glycoprotein gp70 and to the membrane-associated protein p15E. Simultaneously, formation of these mature virion proteins ceases. In addition, lactoperoxidase-catalyzed surface labeling with 125I--iodine indicates that the plasma membrane of cells infected with ts-26 becomes depleted of gp70 antigens at 39 degrees C. Nevertheless, at 39 degrees C these cells release defective MuLVs which lack gp70 and p15E but contain an outer membrane. The released particles also contain an aberrantly processed form of the major virion core protein p30, and many of these virion cores have an unusual immature crescent shape. It has previously been reported that cells infected with the ts-26 mutant of R-MuLV process a 65,000 dalton precursor (Pr65gag) of the virion core proteins more slowly at 39 degrees C than do cells infected with wild-type virus (Stephenson, Tronick and Aaronson, 1975). Although we have confirmed these results, this effect is relatively small and it is known that various alterations of MuLV assembly can lead secondarily to inhibited processing of Pr65gag. We propose that the ts-26 mutant has a primary temperature-sensitive defect in membrane glycoprotein synthesis and that this change causes pleiotropic effects on core morphogenesis.


Utilization of promoter and terminator sites on bacteriophage T7 DNA by RNA polymerases from a variety of bacterial orders. The transcriptional properties of bacterial RNA polymerases purified from seven different species and representing a variety of bacterial orders have been studied using the well characterized DNA from phage T7 as template. The subunit composition of the different preparations suggests that each RNA polymerase holoenzyme has a promoter structure (betabeta'alpha2sigma) similar to that of the well studied E. coli and B. subtilis enzymes. Each enzyme utilizes DNA from bacteriophage T7 as an effective template for RNA synthesis, although all preparations contain a substantial fraction of inactive enzyme. Electrophoretic analysis of the RNA products made with the different RNA polymerases in vitro using T7 DNA (deletion mutant deltaD111) as template reveals that with minor exceptions, all of the heterologous RNA polymerases utilize the same collection of promoter sites on T7 used by the E. coli host enzyme, and only those promoter sites. The T7 early terminator is also efficiently utilized by each enzyme. Since the different bacterial species from which the RNA polymerases were derived are genetically quite distant, it appears that there is a structural element in the promoter which governs its recognition and which is universally recognized among RNA polymerases of different bacterial species. While the different bacterial RNA polymerases generally utilize the same set of T7 promoter sites, the efficiency of utilization of the different promoters varies considerably for different RNA polymerases and for different reaction conditions with the same RNA polymerase. Hence although the several T7 promoter sites share the ability to be recognized by bacterial RNA polymerases, each shows a unique pattern of utilization and therefore must possess a unique element of promoter structure as well. It has previously been shown (Stahl and Chamberlin, 1977) that T7 promoters A1, C, D and E interact differently with E. coli RNA polymerase as judged by the properties of complexes formed between each promoter and the latter enzyme. Since competition takes place among different promoter sites on a template and since these sites can differ functionally, small changes in reaction conditions or in the structure of the RNA polymerase can lead to significant changes in the rate of utilization of different promoter sites even when these promoter sites share common elements. Because the T7 promoters and terminator are utilized efficiently by such a wide range of RNA polymerases and because each of the several T7 promoters possesses unique properties which govern its utilization by RNA polymerase, analysis of the transcripts formed on a T7 DNA template provides a simple and rapid procedure for detecting and analyzing alterations in bacterial RNA polymerases which affect promoter or terminator recognition or utilization.


Prevalence of mitral valve prolapse in presumably healthy young men. We determined the prevalence of mitral valve prolapse (MVP) in presumably healthy young men by studying 107 male house officers and medical students with cardiac auscultation in the supine, sitting and standing positions. Echocardiograms were performed at rest in the supine position before and after amyl nitrite inhalation and were obtainable in 101 subjects. Eleven of the 101 subjects had abnormal findings on auscultation: four had an isolated click and seven had a click and late systolic murmur. Correlation of the independent auscultatory and echocardiographic data in the 101 subjects showed that all seven of the subjects with a click and a murmur had echocardiographic evidence of prolapse. None of the 90 subjects with normal auscultation or the four with an isolated click had an abnormal echocardiogram. All seven subjects with MVP had thoracic skeletal abnormalities, but only one was symptomatic. These data suggest that the prevalence of MVP in healthy young males is similar to the reported 6-10% prevalence in healthy young females.


Echocardiographic assessment of cardiac anatomy and function in hypertensive subjects. Cardiovascular complications are a major source of morbidity and mortality in hypertensive patients. To assess the prevalence of anatomic and functional abnormalities of the heart in such patients, we studied 234 asymptomatic subjects with mild-to-moderate systemic hypertension by echocardiography. After adjusting the echocardiographic values for age and body surface area, we found abnormally increased ventricular septal and/or posterobasal free-wall thickness in 61% of the hypertensive subjects. We found increased left atrial, aortic root, and left ventricular internal dimension (at end-diastole) in 5-7%, and decreased mitral valve closing velocity (E-F slope) and left ventricular ejection fraction were noted in six and 15% of the subjects, respectively. Four percent of the patients had disproportionate septal thickening (i.e., ventricular septal-to-left ventricular free-wall thickness ratio greater than or equal to 1.3). In contrast to the high prevalence of cardiac abnormalities detected by echocardiography, less than 10% of the hypertensive subjects had abnormal 12-lead ECGs or abnormal chest x-rays. These findings demonstrate a high prevalence of cardiac abnormalities in a population of asymptomatic hypertensive subjects. These abnormalities can be detected by echocardiography before they are otherwise apparent.


Carotid baroreflex function in young men with borderline blood pressure elevation. Carotid baroreflex function was assessed in 10 normotensive young men and 20 age-matched subjects with borderline hypertension (successive blood pressures above and below 140/90 mm Hg) by measuring sinus node responses to brief neck suction. Subjects with borderline hypertension were divided into two equal groups according to their average systolic arterial pressures. Baroreflex responses were reset to function at higher pressure levels than normal in subjects with mild borderline hypertension, but reflex sensitivity was normal. Responses were also reset in subjects with more severe borderline hypertension, but reflex was subnormal. The results suggest that a gradation of baroreflex responsiveness exists among patients classified as having borderline hypertension: Subnormal responsiveness was found in those subjects whose resting average systolic arterial pressure was greater than or equal to 140 mm Hg.


Comparative assessment of stimuli that release neuronal and adrenomedullary catecholamines in man. We assessed the release of neuronal and adrenomedullary catecholamines in response to various stimuli of the sympathetic nervous system in normal subjects. Plasma catecholamines and their urinary metabolites, normetanephrine and metanephrine, were measured. Sodium restriction increased supine plasma norepinephrine by 37% and ambulatory plasma norepinephrine by 22%, with urinary normetanephrine excretion increased 29%. The sodium restriction did not elevate plasma epinephrine or urinary metanephrine. The most potent stimuli of norepinephrine were treadmill exercise, orthostasis, caffeine, the cold pressor test, sodium restriction and handgrip exercise, in descending order. Plasma epinephrine was increased by caffeine, treadmill exercise, the cold pressor test, handgrip exercise and the Valsalva maneuver, in that order. Syncope resulted in profound changes in plasma epinephrine but only modest changes in plasma norepinephrine. We conclude that in man, there is frequent dissociation between the effects of different stimuli on neuronal and adrenomedullary catecholamine release.


Effects of volume expansion and contraction in normotensive whites, blacks, and subjects of different ages. We studied the blood pressure, natriuretic, kaliuretic and humoral responses of 347 normal subjects after volume expansion and volume contraction to examine possible differences among whites, blacks and subjects of different ages. According to outpatient 24-hour urine collections, blacks excreted less sodium and potassium than whites. After similar states of sodium intake were achieved among all subjects, 2 liters normal saline were given i.v. Blacks and subjects greater than or equal to 40 years excreted less sodium than whites or subjects less than 40 years, over a 24-hour period. In addition, blacks excreted less potassium. The delay in sodium excretion occurred during the first 12 hours after the salt load. Blacks had a greater suppression of plasma renin activity than whites 24 hours after saline. Blacks also had higher blood pressures than whites after saline administration; their pressure remained elevated until furosemide was given. Furosemide, 120 mg over 24 hours, evoked greater natriuresis, but less kaliuresis in blacks than in whites. The greater prevalence of hypertension in both blacks and older subjects may be related to relatively blunted natriuretic responses when these groups engage in the high sodium-low potassium intake characteristic of our society.


Cardiovascular response to isometric exercise in normal adolescents. The purpose of this study was to document the cardiovascular response to submaximal isometric handgrip (IHG) exercise in 32 normal adolescents. Left ventricular (LV) dimensions and systolic time intervals were recorded using echocardiography; blood pressure was measured by sphygmomanometry both at rest and during IHG exercise at 25% maximum contraction. This level of isometric exercise produced significant (p less than 0.001) increases in mean heart rate, systolic, diastolic and mean blood pressures. Despite this response LV diastolic and systolic dimensions remained unchanged during exercise; hence, stroke volume remained constant. Cardiac index increased by 22% (p less than 0.001) due to the increase in heart rate. Systemic vascular resistance did not change significantly. LV ejection indices, including shortening fraction, mean Vcf and systolic time intervals also remained unchanged, except for an increase in LV ejection time index (p less than 0.025). These data indicate that the cardiovascular response to submaximal isometric exercise in normal adolescents is similar to that reported in adults. This study demonstrates that sustained isometric stress testing in adolescents is safe and provides normal hemodynamic values.


The characteristic sequence for the onset of contraction in the normal human left ventricle. The sequence for the onset of segmental contraction of the left ventricle was studied in 25 normal patients by analyzing sequential frames obtained at 16.7-msec intervals of right anterior oblique (RAO) ventriculograms by two independent methods. In the first method, we compared the times of onset of contraction of the hemidiameters associated with each of 54 segments with time of onset of contraction of the average of all the hemidiameters for the ventricular contour. In the second method we used a radial coordinate system and determined relative phase relationships by plotting the motion of each of 54 segments against the average motion of all segments. The resulting pattern showed that, on the average, the midregion of the inferior wall began to contract 25 msec before the apex and the midregion of the anterior wall began contraction 18 msec before the apex. In 12 of 25 patients the interior and anterior walls both began to contrast before the apex. In only one of 25 patients did the apex begin to contract first. This sequences of contraction corresponds to the reported sequence of electrical activation for normal human left ventricles.


Thallium-201 myocardial scintigraphy in patients with triple-vessel disease and ischemic exercise stress tests. Thirty patients with triple-vessel coronary artery disease proven by angiography, symptomatic angina and a positive ECG stress test were evaluated with thallium-201 (201TI) scintigraphy. Twenty patients also had aortocoronary saphenous vein bypass surgery; 15 of them had repeat noninvasive evaluation. Seventy percent of these patients showed ischemia by 201TI scintigraphy, of which one-half returned to normal after surgery. Postoperative reversion of the ECG stress test together with 201TI stress/reperfusion imaging correlated well with the completeness of surgical revascularization. We could not explain the prevalence (80%) of infarcts detected by 201TI in this group, of which 76% could be anatomically correlated to epicardial scars. The positivity of infarcts by 201TI exceeded that predicted by previous history of infarction, Q waves on resting ECG or ventriculographic akinesis. These observations suggest that 201TI scintigraphy is a useful noninvasive tool in the follow-up and understanding of patients with coronary heart disease. These conclusions also support the concept that 201TI stress imaging need not have the identical connotation as the ECG stress test.


Aspirin-induced increase in collateral flow after acute coronary occlusion in dogs. Aspirin (acetylsalicylic acid) has inhibitory effects on platelet function and prostaglandin synthesis. Since alterations in either platelet function or prostaglandin-mediated vascular responses could influence blood flow to ischemic myocardium, we tested the effects of aspirin on coronary collateral flow after acute occlusion of the left anterior descending coronary artery in dogs. Aspirin dose (600 mg i.v.) consistently inhibited in vitro ADP-induced platelet aggregation. In 13 open-chest dogs, regional myocardial blood flows (radioactive microsphere technique) were determined at 5 minutes and 4 hours after occlusion). In seven of these dogs, aspirin (600 mg i.v.) was administered 1 hour before occlusion. In the aspirin-treated dogs, collateral flow increased significantly (p less than 0.05), from 0.09 +/- 0.02 ml/min/g at 5 minutes to 0.15 and 0.02 ml/min/g 4 hours after occlusion. Collateral flow was not significantly altered over 4 hours in control dogs. The aspirin-induced increase in collateral flow was confined to epicardium (12 +/- 4% of normal zone flow at 5 minutes to 23 +/- 4% at 4 hours after occlusion).


Aberrant coronary artery origin from the aorta. Report of 18 patients, review of literature and delineation of natural history and management. To clarify the natural history and management of patients with aberrant origin of a coronary artery from the aorta, we reviewed 18 patients whose right (RCA) or left (LCA) coronary artery arose aberrantly and passed between the aorta and right ventricular infundibulum. Nine patients had aberrant LCA. Three young males died suddenly after exertion, each with proximal focal LCA stenosis. None of the six adults with angina (ages 36-70 years) studied angiographically had proximal LCA stenosis. Unlike the young, "sudden death-prone" patients in whom coronary bypass of proximal stenosis may have prophylactic value, our older patients with aberrant LCA did not have proximal stenosis or sudden death, and therefore LCA bypass for sudden death prophylaxis is not warranted. Nine patients (ages 18-60 years) had aberrant origin and course of the RCA. Seven patients studied angiographically because of angina had no focal proximal RCA stenosis, but two patients had hypoplastic RCA ostia. Although a potential concern in these latter patients, sudden death has not been reported with aberrant RCA, therefore in the absence of syncope, RCA bypass for sudden death prophylaxis is not indicated.


Ventricular premature contractions: a randomized non-drug intervention trial in normal men. The influence of a 6-week intervention on factors thought to be related to ectopic cardiac rhythms was tested in normal men with frequent ventricular premature contractions (VPCs), using a randomized, controlled and partial crossover design. The VPC intervention trial experimental regimen included total abstinence from caffeine and smoking, reduction of alcohol intake, and a physical conditioning program. Effects were studied in detail among 81 healthy men with persistent VPCs. VPCs were measured during standard states of rest, dynamic and isometric exercise and other stresses, and 24-hour ambulatory monitoring. Adherence to the treatment was excellent. The experimental group achieved more than 80% of activities asked of them, and little "contamination" occurred in the control group. VPCs were analyzed according to VPC/min, VPC/man and VPC/total number of heart beats. Moderate changes in VPC rates occurred in both experimental and control groups but no significant group differences were found at rest or during any induction test. This 6-week, multiple-factor "hygienic" intervention program had no significant influence on the frequency or occurrence of VPCs in apparently normal men with persistent and frequent VPCs. Because the mechanisms and the significance of VPCs are different in patients with ischemic heart disease, our approach and methods may be useful for similar trials among cardiac patients of adjunct or non-drug therapy for ectopic rhythms.


Preeminence of the left stellate ganglion in the long Q-T syndrome. In seven patients with Romano-Ward syndrome, stellate ganglion block or stimulation and pharmacologic interventions were made to assess their influence on duration of the O-T interval, electrical alteration of the T wave and ventricular tachydysrhythmias. Left stellate ganglion block and right stellate ganglion stimulation shortened Q-T interval, abolished alternans phenomena and suppressed tachydysrhythmias. Propranolol and phenytoin had a similar effect. In contrast, right stellate ganglion block, left stellate ganglion stimulation and prior administration of quinidine and procainamide had an opposite effect. These responses resemble observations in animal models which suggest that excessive or unopposed activity of the left, or subnormal activity of the right stellate ganglion, or both, account for the pathophysiologic manifestations of the long Q-T syndrome. They are also consistent with clinically correlated, cardiac neuropathologic findings in these patients. An analogous but acquired dysautonomia involving the left stellate ganglion and ischemic left ventricle may precipitate sudden coronary death.