The influence of stress and stress hormones on the transplantability of a non-immunogenic syngeneic murine tumor. Quantitative transplantation assays of a syngeneic murine adenocarcinoma have been used to investigate the effects of stress hormones and tumor take probability. Cortisol, injected intraperitoneally one hour before and 3 hours after tumor cells, caused a dose dependent reduction of TD50 (number of tumor cells required for 50% takes) by factors of from 4 at a total dose 4 microng/g to 68 at 400 microng/g. ACTH, given at 0.2 I.U. daily for 9 days spanning the time of tumor cell injection, reduced the TD50 2.5-fold, indicating that the peak gluco-corticoid level achieved, rather than its duration, was of greater significance. Adrenaline, while much less effective than cortisol, produced an 8-fold reduction in TD50 at its maximum tolerable dose. The effect of cortisol simulated that of whole body irradiation (WBI), and while both these agents depress immune reactivity, evidence is presented to suggest that immunological mechanisms are not responsible for their effect. WBI constitutes a systemic stress, and the demonstration that surgical trauma (laparotomy) could also reduce the TD50 for this tumor suggested that both might act via endogenous glucocorticoids. However, the failure of prior total adrenalectomy of mice to abrogate the effect of either WBI or laparotomy indicated that stress hormones were not essential intermediaries. It is concluded that both stress hormones, especially glucocorticoids, and stressful procedures acting independently of stress hormones, can facilitate tumor transplantation.


Correlative study of the morphology and C19-steroid metabolism of benign and cancerous human prostatic tissue. Perineal punch biopsy specimens of human prostate with benign hyperplasia (BPH), well- and poorly-differentiated adenocarcinoma and transitional-cell carcinoma were incubated with testosterone-1,2-3H and 5alpha-dihydrotestosterone-1,2-3H. Incubations were carried out using a single tissue-radiosubstrate ratio and time point. Resulting radiosteroid patterns were related to histologic and ultrastructural features of these tissues. Well differentiated neoplasms had ultrastructural characteristics closely resembling hyperplastic epithelia. Both in BPH and in well differentiated carcinomas the C19-steroids were mainly metabolized by the 17beta-hydroxysteroid pathway. In contrast, cells in poorly-differentiated adenocarcinoma and transitional-cell carcinoma lacked the cytoplasmic organelles responsible for secretion; formation of 5alpha-reduced 17beta-hydroxysteroids was decreased in these carcinomas, while conversion to 17-oxosteroid radiometabolites remained unchanged or was greatly increased. These results indicate that loss of prostatic differentiation is attended by a trend from reductive toward oxidative radiotestosterone metabolism. Even in NAPDH-supplemented preparations of the majority of poorly-differentiated tumors, there was diminished transformation to 5alpha-dihydrotestosterone, the key intracellular hormone in the expression of androgenic activity in the prostate. These findings may explain why poorly-differentiated prostatic neoplasms are frequently unresponsive to anti-androgenic therapy.


Malignant granular cell tumor of the radial nerve: an autopsy observation with electron microscopic and tissue culture studies. Malignant granular cell tumor closely associated with the radial nerve in the left upper arm, was found in a 34-year old man. The tumor increased rapidly in size, was locally invasive and showed recurrence and metastases to lymph nodes and the lungs. The tumor was composed solely of granular cells and its histological features were very similar to those of benign granular cell tumors. However, considerable variation of cellular size and shape, hyperchromatic plump nuclei, and some disordered cellular arrangement were thought to be significant indications of malignancy. Ultrastructural study of the tumor revealed many lysosome-like dense granules in the cytoplasm. The presence of a few axon-like cytoplasmic processes between granular cells and the intimate anatomical association of the tumor with the radial nerve seemed to support the concept of peripheral nerve origin of the malignant granular cell tumor.


Uterine leiomyoblastoma: ultrastructural and cytological studies. A case of uterine leiomyoblastoma is presented with ultrastructural and cytological findings. As in gastric leiomyoblastoma, fine filaments of dimensions consistent with myofilaments and marginal dense bodies are present within the tumor cells, providing evidence for the smooth muscle origin of this neoplasm. Numerous cytoplasmic vacuoles are seen in cellular samples obtained by directly scraping the lesion corresponding to the vacuoles present in glutaraledehyde-fixed material examined by electron microscopy. They do not appear to be artifacts of poor fixation and may be related to the nature of the tumor. Cytologically the cells do not resemble epithelial cells of either squamous or glandular origin and are more consistent with cells of mesenchymal origin.


Rarity of cancer of the cervix in the Malaysian Orang Asli despite the presence of known risk factors. Cancer of the cervix is exceedingly uncommon in the Malaysian Orang Asli (aborigine), despite the presence of factors associated with an increased risk of developing this malignancy. In only three patients was the diagnosis of carcinoma of the cervix established, out of a total of nearly 18,000 female inpatients, admitted to the Gombak Orang Asli Hospital over a 13-year period. Over this same period, 81 female patients were diagnosed as having cancer. Interviews with female Orang Asli patients show the presence of alleged risk factors for cervical cancer, including early age of first intercourse, multiparity and non-circumcision of husbands. The low incidence of cancer of the cervix in this aborigine community may be due to the strict moral code of the Orang Asli, limiting extramarital sexual activity and associated venereal infection.


Microfollicular thyroid carcinoma with amyloid rich stroma, resembling the medullary carcinoma of the thyroid (MCT). A human thyroid tumor is described which, on light microscopy, exhibited the features of medullary carcinoma of the thyroid (MCT). The cells were arranged in solid lobules or trabeculae, and the stroma was abundant and gave positive reaction for amyloid as assessed by congo red, crystal violet and thioflavin T stains. However, there was microfollicular differentiation in certain portions of the tumor even where the tumor was invasively growing. On electron microscopy, numerous microfollicles were identified even where unsuspected by light microscopy. There were junctional complexes, the cells possessed well developed rough endoplasmic reticulum, prominent Golgi and numerous dense bodies obviously derived from the Golgi vesicles, which were of the same morphology as "secretory" granules described previously by some authors in MCT. However, we find this kind of cytology typically in microfollicular thyroid carcinomas. Presence of about 100 A thick fibrils in the stroma was consistent with histochemically positive amyloid. The biochemical data were compatible with the differentiated follicular cell origin of the tissue. The homogenate contained poorly iodinated thyroglobulin, and thyroid peroxidase activity. Calcitonin was undetectable by a sensitive radioimmunoassay. It is concluded that this tumor was follicular thyroid carcinoma with amyloid rich stroma. The presence of amyloid and dense bodies with homogenous electron dense contents is insufficient for making conclusions about histogenesis of thyroid tumors. The so called MCT with amyloid stroma probably represents a heterogenous group of thyroid tumors, at least some of them derived from follicular epithelium.


Peripheral ameloblastoma: a study of 21 cases, including 5 reported as basal cell carcinoma of the gingiva. The peripheral ameloblastoma and the basal cell carcinoma of the gingiva are probably the same lesion. This article describes the clinical features, natural history, and histopathology of nine acceptable cases published as peripheral ameloblastoma, five lesions published as basal cell carcinoma of the gingiva, and seven  unpublished cases of peripheral ameloblastoma. Tissue sections were studied from seven of the cases published as peripheral ameloblastoma, three of the cases published as basal carcinoma, and all the unpublished cases. The peripheral ameloblastoma may exhibit several of the various histologic patterns found in the intraosseous ameloblastoma but has a marked tendency to be acanthomatous. These lesions appear to arise from either remnants of the dental lamina within the gingiva or from the surface epithelium. They are relatively innocuous lesions lacking the persistent invasiveness of intraosseous ameloblastoma. Peripheral ameloblastomas should be excised with a small margin of normal tissue and the surgical site re-examined periodically.


The morphology of human papillomas of the upper respiratory tract. Recurrent squamous papillomas of the upper respiratory tract were examined by light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Surface of the cells is irregular and is covered by numerous stout microvilli. These are shorter and broader than those of cells of the uninvolved mucosa. The villi often seem umbilicated at the apex; the remainder of them, however, are rounded. The epithelium participating in the formation of papillomas shows some maturation of the cells but this does not progress normally. The predominating area is the thickened spinous layer representing the bulk of the lesion. The basal layer shows mildly increased activity but the basement membrane is intact. The cells often are very closely packed but in some areas, more particularly  in the deep layer, they are loosely arranged. The intercellular space contains a moderately electron-dense finely fibrillar material. No abnormal mitoses are found. The neighboring univolved epithelium often shows increased growth activity and some inflammation. The larygneal papillomas propably represent an overgrowth of epithelium which may develop following hindered desquamation caused and/or heralded by a chronic inflammatory condition probably of viral origin and may be preceded by epithelial metaplasia and hyperplasia.


Amylase in the lung.

Amylase in the lung. Although elevated amylase levels in serum, pleural fluid, and extracts of tumor tissue in primary lung cancer have been reported, electrophoretic and column-chromatographic studies have not revealed the ectopic production of amylase but have merely shown an increase of amylase activity of chiefly the salivary type in these materials. The present study was designed to make clear the nature of the amylase or amylase-like substance in the serum, pleural fluid and tumor extracts, and to determine whether amylase might be produced ectopically in tumor tissues. Our data not only confirmed that the hyperamylasemia in some cases of primary lung cancer was due to an increase in salivary type isoamylases, but also showed that the same isoamylase pattern occurs in serum, pleural fluids, and diseased lung tissue of patients with pneumonia. However, the elution pattern of amylase in these materials in column-chromatography on Sephadex G-75 Superfine was different from that of salivary amylase. On the basis of our observations, it seems reasonable to conclude that the salivary type hyperamylasemia in some cases of primary lung cancer may be due to an increase in the amylase contained in normal lung tissues, resulting from activation and release into the blood stream by some inflammatory process. However, ectopic production of amylase was demonstrated in one particular case of primary lung cancer in which a high amylase content and a peculiar isoamylase were found both in the primary and metastatic lesions.


Squamous cell carcinoma of the buccal mucosa associated with chronic oral polyvinyl chloride exposure: report of a case. The following case report describes a 22-year-old healthy man, non-smoker, non-drinker, who developed a squamous cell carcinoma of the buccal mucosa. The appearance of this unusual tumor in an unexpected clinical setting was unexplained until it was discovered that the patient had a lifetime habit of chewing plastic materials containing polyvinyl chloride (PVC). The extent and chronicity of his intense oral exposure to PVC is believed to be related to his subsequent development of an oral neoplasm. Attention is again focused on PVC as an important environmental toxin and carcinogenic agent. Clarification of the content of PVC in consumer products is urged. Physicians must remain aware of possible environmental carcinogens in patients presenting with unexpected or rare malignancies. Detection of such exposure may well prevent subsequent development of additional unexplained neoplasms.


Ectopic production of human placental lactogen by human breast tumors. Serum samples from 72 patients with established carcinoma of the breast were investigated for ectopic presence of hPL. Further, the relationship of ectopically-secreted hPL and hCG-beta in breast cancer was investigated. Ten of 72 patients examined had detectable hPL and 12 had detectable hCG-beta, at 1-2 ng/ml serum sensitivity of the assay. The presence of hPL in serum of breast cancer patients was found to be independent of that of hCG-Beta. Sera of 13 patients with cystic mastitis, five with fibroadenoma, two with acute inflammation of breast, 20 normal women (non-pregnant) and 20 normal men did not show any detectable serum hPL or hCG-beta at the above mentioned sensitivity of the assay. Since these hormones were not detectable in normal men, normal non-pregnant women, and in patients having other pathological conditions of breast, the possible use of them as markers in cancer is expected


The relationship of fever, granulocytopenia and antimicrobial therapy to bacteremia in cancer patients. The relationship of fever, granulocytopenia, and antimicrobial therapy to bacteremia was studied retrospectively in 53 cancer patients. Severe granulocytopenia was present at the time blood cultures were positive in 27 to 31 episodes of bacteremia. Twenty-five episodes of bacteremia documented before the initiation of antimicrobial therapy in patients who were granulocytopenic and febrile. No bacteremia occurred in the absence of fever. Only two bacteremias occured while patients were receiving parenteral antimicrobials. Antimicrobial therapy was terminated 30 times in the presence of granulocytopenia and fever, and subsequent bacteremia occurred in 14 patients within 4 days. Patients who died with fungal disease did not receive more antibiotics than patients who died without fungal disease. These data suggest a rationale for long-term use of antimicrobial therapy in patients with persistent granulocytopenia and fever.


Barium enema, proctosigmoidoscopy and upper gastrointestinal series in the preoperative evaluation of the cancer patient. Charts of 566 patients admitted to the Surgery Branch, National Cancer Institute over a five year period were reviewed. Routine upper gastrointestinal series was performed in 453 patients; barium enemas in 490; and proctosigmoidoscopies in 342. Upper gastrointestinal series detected no metastases or second primary malignancies but delineated the extent of large intra-abdominal tumors. Barium enema and proctosigmoidoscopy together revealed three unsuspected primary colonic tumors (1.2% in patients over 50 years of age). Sigmoidoscopy and examination of the stool for occult blood would have detected the same patients. Barium enema may be limited to surgical cancer patients in whom the extent of local tumor invasion is to be defined and to older patients in whom colonic tumors are suspected. Neither UGIS nor barium enema appear to be of value as a routine preoperative screening test in surgical cancer patients.


A comparison of Hodgkin's disease in Alameda County, California, and Connecticut: histologic subtype and age distribution. A comparison of the histologic patterns of Hodgkin's disease in Alameda County, California and Connecticut was made in relation to age. All Hodgkin's disease cases from the Alameda County Cancer Registry diagnosed between 1960 and 1969 for which adequate material was available for evaluation were reviewed and histologically subtyped according to the Rye classification (159 cases). The age-specific relative frequencies and incidence rates for the histologic subtypes of Hodgkin's disease in Alameda County were estimated and compared to those estimated for Hodgkin's disease in Connecticut. The morphologic ex pression of Hodgkin's disease in both areas was similar and characteristic of economically developed regions with high living standards. The comparison supports the hypothesis that the host immune capacity, influenced by socioeconomic factors, has a strong and measurable effect on the pathogenesis of Hodgkin's disease.


Follow-up of survivors of childhood cancer. A registry has been established of survivors of childhood cancer treated at the Sidney Farber Cancer Institute, Boston. This resource provides opportunities for studies of physiologic, genetic, psychologic, and social sequelae of childhood neoplasia. Findings indicate potential in most patients for a high quality of extended life. Comprehensive long-term care may further reduce late morbidity and mortality.


Cancer risk as related to use of oral contraceptives during fertile years. A case-control study of 452 breast cancer patients, aged less than 50 years, and 872 age-, race-, and religion-matched control patients generated relative risk estimates of breast cancer associated with oral contraceptive practices. The relative risk of breast cancer from ever-use of oral contraceptives was 1.1, not significant. Relative risks did not differ by age, interval since first use, interval since last use, or time periods in which steroid compounds differed in composition and potency. However, the relative risks of breast cancer from current use, from 2 to 4 years of ever-use, from 6 or more years of use by women with prior benign breast disease, and from use before first childbirth were increased significantly. The findings suggest the malignant process may be quickened if transformed cells are present during oral contraceptive use. Yet, the findings neither indicate that oral contraception induces breast cancer nor do they exonerate female steroid hormones. The findings do encourage continued surveillance of steroid contraception for cancer induction or promotion. In addition to duration of oral contraceptive use and other measures of dose response, future observations should pay especial attention to use by women before first childbirth and by women with already established benign breast disease.


Epidemiology of vaginal adenosis and adenocarcinoma associated with exposure to stilbestrol in utero. Following the observation that maternal ingestion of diethylstilbestrol was associated with the development of adenocarcinoma of the vagina in young women, analysis of 170 cases of the disease, collected over a two-year period, was undertaken. Details of history of non-steroidal estrogenic intake during pregnancy, its dosage and duration of intake and prevalence of concomitant abnormalities of the vagina and cervix were recorded. This analysis disclosed that dosage and duration of intake do not appear to be vital factors in contributing to the development of carcinoma. A controlled, prospective investigation of prenatal exposure to stilbestrol was also carried out. Vaginal and cervical abnormalities were common among 110 young women exposed to stilbestrol in utero. Biopsy-proved vaginal adenosis was present in 35% of the exposed population as compared with only 1% of the control subjects. Fibrous ridges of the vagina and cervix and failure of the vaginal mucosa and portions of the cervix to stain with iodine were also more frequently observed in the exposed population. While the risk of developing adenocarcinoma appears to be very low, a majority of exposed females do have other abnormalities of the lower genital tract. While some investigators believe that adenosis is a premalignant lesion, at present there is no clear-cut evidence of transition from adenosis to adenocarcinoma.


Oral contraceptives and breast neoplasia. The continued high prevalence of oral contraceptive (OC) use by young women warrants the careful study of these agents. At present, it appears that the risk of benign breast disease is reduced by use of OCs. This effect seems to be greater for fibrocystic disease than for fibroadenoma. Available information suggests that, as yet, OC use has not had any effect on breast cancer risk. The recent slight rise in breast cancer rates in younger women has not been shown to be due to OC use. The relationship between OCs and breast cancer may not become clear for another 5 to 10 years because of the long latent period of this disease. Meanwhile, for reasons presented, it would seem wise not to prescribe OCs for women with a history of benign breast disease and to withdraw them from women who develop these conditions.


Induction of preneoplastic and neoplastic lesions in grafted rat tracheas continuously exposed to benzo(a)pyrene. Heterotopically transplanted rat tracheas were continuously exposed to measured amounts of benzo(a)pyrene over a period of 1 to 6 months. The cumulative doses ranged from 10 to 2490 microng. The morphological response of the tracheal epithelium was characterized by hyperplasis during the first 2 weeks, followed by atrophy. Squamous metaplasias did not appear until after 4 months of exposure; at 4 and 6 months numerous dysplastic lesions and noninvasive carcinomas resembling those seen in the airways of humans were found in the higher carcinogen dose groups. The first invasive carcinomas developed at 4 months in the groups given 1250 microng or more benzo(a)pyrene. The lowest dose tested that produced a carcinoma within the observation period of 22 months was 300 microng benzo(a)pyrene. The majority of the neoplasms were squamous cell carcinomas, although several adenocarcinomas and sarcomas also developed. Since a variety of metaplastic and dyplastic lesions can be induced by carcinogenic polycyclic hydrocarbons in the transplanted rat tracheas, this experimental model appears to be well suited for the study of the sequential epithelial changes that lead to respiratory tract neoplasia.


Fluidity of membrane lipids and lateral mobility of concanavalin A receptors in the cell surface of normal lymphocytes and lymphocytes from patients with malignant lymphomas and leukemias. Lymphocytes isolated from the peripheral blood of patients with nonmalignant and malignant disorders were studied for fluidity of membrane lipids and lateral mobility of concanavalin A (Con A) receptors. The degree of fluidity of the surface membrane lipid core was monitored quantitatively by fluorescence polarization analysis using the probe 1,6-diphenyl-1,3,5-hexatriene embedded in lipid regions of the surface membrane of intact cells. Mobility of Con A surface receptors was determined by the cap-forming ability after binding of fluorescent Con A. The present studies were performed on lymphocytes from 28 patients with malignant lymphomas, 22 patients with leukemia, 28 individuals who either were healthy or had nonmalignant disorders, and 5 patients with carcinoma. The results showed that lymphocytes and mononuclear cells from patients with malignant lymphomas and leukemias have a more fluid lipid layer in their surface membrane than do lymphocytes obtained from healthy individuals or from patients with other malignant and nonmalignant disorders. This increase in membrane fluidity was less pronounced in lymphocytes isolated from leukemic patients in clinical remission and from leukemic patients receiving treatment with steroids. The results also show a marked difference in the cap-forming ability of lymphocytes from patients with malignant lymphomas or leukemia as compared with lymphocytes from patients with non-malignant disorders or carcinoma. Lymphocytes isolated from lymphoma and chronic lymphatic leukemia patients during remission stages of the disease exhibited a higher cap-forming ability. The cap-forming ability of cells from patients with chronic lymphocytic leukemia was unaffected by treatment with steroids. The present results, which are in line with previous observations, have shown that normal lymphocytes can be characterized by a low degree of lipid fluidity but a high degree of mobility of Con A receptors, whereas leukemic lymphocytes are characterized by a high degree of lipid fluidity but a low degree of mobility of Con A receptors. These results confirmed our general hypothesis on the dynamic interrelation between membrane lipids and membrane protein receptors, and they indicate that the widely accepted term "membrane fluidity" requires better consideration for different membrane components.


Morphology, growth, chromosomal pattern and fibrinolytic activity of two new human neuroblastoma cell lines. Neuroblastoma cell lines LA-N-1 and LA-N-2 were extablished from neuroblastoma cells in the bone marrow and in the primary tumor, respectively, of two children with metastatic neuroblastoma. Morphology, growth in vitro and in athymic nude mice, chromosomal patter, and fibrinolytic activity of these cell lines and of previously extablished human neuroblastoma cell lines IMR-32, SK-N-MC, and SK-N-SH were compared. Most LA-N-1 cells were tear-drop shaped, small cells with processes; they tended to grow in clusters. LA-N-2 was comprised of elongated cells and small round cells, the latter growing in dense clumps on the former. Electron microscopy revealed numerous cytoplasmic dense cores in many LA-N-1 cells but none in LA-N-2 CELLS. During logarithmic growth in vitro, doubling times for LA-N-1, LA-N-2, SK-N-MC, SK-N-SH, and IMR-32 cells were 32,56, 23, 36, and 26 hr, respectively. Cells of all lines formed colonies in soft agar, and, after variable latency periods, LA-N-1, LA-N-2, SK-N-MC, and IMR-32 cells formed tumors in athymic nude mice. The marker chromosome(s) characteristic of each cell line was present in more than 90% of cells of given line. Significant plasminogen-dependent fibrinolytic activity was present in cells of all lines. These studies indicate that LA-N-1 and LA-N-2 cells arose from single but different aberrant progenitor cells and that they have properties of neuroblastoma cells. They also demonstrate that cell lines derived from human neuroblastomas are heterogenous as are the tumors in children.


Plasma pharmacokinetics of adriamycin and its metabolites in humans with normal hepatic and renal function. A new, nondestructive, plasma extraction technique ultilizing chloroform:isopropyl alcohol (1:1) and ammonium sulfate saturation has been devised to isolate adriamycin and its metabolites from human plasma. Adriamycin was the most prominent species in plasma. It disappeared according to  a triphasic pattern with a mean half-life of 30 hr. Six metabolites have been clearly separated from adriamycin by thin-layer chromatography. Three were aglycones and three were polar metabolites, one of which has been identified as adriamycinol. All metabolites appeared rapidly in plasma and disappeared according to a biphasic or tri-phasic pattern. The polar metabolites in plasma were found in similar relative concentration to those in urine. In contrast to the small Quantities of aglycones in urine, however, significant concentrations of aglycones were found in plasma. The least prominent metabolite was adriamycin aglycone; the most prominent metabolite was a less polar aglycone, most likely deoxyadriamycin aglycone, and a more polar aglycone, presumably demethyl deoxyadriamycinol aglycone, was the only metabolite to show variable pharmacokinetics in different patients. The nondestructive plasma extraction technique has verified the presence of extensive human metabolism of adriamycin and demonstrated the presence of aglycone and polar metabolites.


Influence of inducers and inhibitors of mixed-function oxidasts on benzo(a)pyrene binding to the DNA of rat liver nuclei. Benzo(a)pyrene-conjugated DNA was isolated after benzo(a)pyrene was incubated in vitro with liver nuclei from control, phenobarbital-treated, and methylcholanthrene-treated rats and the reduced nicotinamide adenine dinucleotide phosphate-generating system. Aryl hydrocarbon hydroxylase, the levels of activity of which varied with the different nuclear systems, was highly specific in the activation of the benzo(a)pyrene to forms that bind to DNA. Sephadex LH-20 chromatography of the degreded DNA from liver nuclei of pretreated rats revealed an in vivo DNA-bound product  previously shown to be derived from 7,8-dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide (Product A). The nuclei from phenobarbital-pretreated rats also contained a DNA-bound product corresponding to that derived from the binding of benzo(a)pyrene 4,5-oxide (Product C), but they completely lacked the DNA-bound product derived from futher metabolism of 0-hydroxybenzo(a)pyrene (Product D). In contrast, nuclei from methylcholanthrene-treated rats yielded large amounts of Product D. No bound products were found with the control nuclei. 7,8-Benzoflavone caused an 80% inhibition of benzo(a)pyrene binding to DNA in the nuclei of methylcholanthrene-pretreated rats, but it had no effect on control nuclei or those from phenobarbital-pretreated rats. It inhibited formation of Product A, but not Products B or C, in nuclei from phenobarbital-pretreated rats. With nuclei from methylcholanthrene-pretreated rats. With nuclei from methylcholanthrene-pretreated animals, 7,8-benzoflavone led to an overall loss of products, but small amounts of Products A and B persisted. 1,1,1-Trichloropropene 2,3-oxide strongly inhibited nuclear aryl hydrocarbon hydroxylase activity and the phenol fraction measured by thin-layer chromatography in all nuclear systems, but it significantly increased the benzo(a)pyrene binding in all nuclei. With nuclei from phenobarbital-treated rats, it inhibited the formation of Product B but not of Products A and C; whereas with nuclei from methylcholanthrene-pretreated rats, it inhibited the fromation of all the hydrocarbon-deoxyribonucleoside products.


The distribution of [3H]vinblastine in tumor and host tissues of Nb rats bearting a transplantable lymphoma which is highly sensitive to the alkaloid. A new transplantable lymphoma in Nb rats responded dramatically to treatment with vinblastine (VLB). A single i.p. injection of VLB, 0.8 mg/kg, caused even highly advanced tumors to regress until they were no longer palpable. or investigation of the hypothesis that the oncolytic response may reflect a special affinity of VLB for the tumor, lymphoma-bearing rats were given an i.p. injection of -e13H]VLB, and the levels of radioactivity and [3H]VLB in the tumor and host tissues were determined as a function of time. Radioactivity was concentrated by the lymphoma relative to the blood (mostly as unchanged [3H]VLB) at levels that showed only a modest decline over a period of at least 48 hr. During this time the [2H]VLB in both the plasma and whit blood cell fraction of the blood declined markedly and continuously to very low levels. Thymus and lymph nodes resembled the lymphoma in showing a long-term retention of radioactivity. The levels of radioactivity in the spleen, liver, and bone marrow were initially much higher than that in the tumor but decreased markedly with time. In addition very little of the radioactivity remaining in the spleen and liver at 48 hr was due to [3H]VLB, and by this time the VLB concentration in these tissues was much lower than in the tumor. It is suggested that the chemotherapeutic response of the lymphoma may be related to the continuing presence of a significant concentration of VLB in this tumor after the plasma VLB had fallen to very low (subantimitotic) levels.


Carcinogenicity and cocarcinogenicity of chromium carbonyl in heterotopic tracheal grafts. Tracheal grafts, implanted s.c. on syngeneic rats, were used as a bioassay for carcinogenicity or cocarcinogenicity of chromium carbonyl (CC). After a period of revascularization, the lumens of 22 grafts were filled with an agar suspension of 2.5 mg of CC. Twenty-two grafts were filled wit a suspension of 2.5 mg of benzo(a)pyrene (BP) and a mixture of the two chemicals was placed in 24 other grafts. Four controls were exposed to the vehicle only. Eight squamous cell carcinomas developed in the tracheas treated with BP alone and 10 similar neoplasms arose in the CC-BP treated group, but 3 of those induced by CC-BP had metastasized by 9 months. CC alone induced carcinomas in two grafts. The data indicate that this metal carbonyl is a carcinogen that can act synergistically with BP and demonstrate the utility of the technique as an efficient tissue-specific bioassay.


Effect of mouse interferon and polyriboinosinic acid-polyribocytidylic acid on L-cell tumor growth in nude mice. L-929 cells injected s.c. into athymic nude mice cause the formation of single well-defined tumors at the site of inoculation. The development of these tumors is markedly inhibited by polyriboinsinic acid-polyribocytidylic acid [poly(l)-poly(C)], whether poly(l)-poly(C) treatment is initiated one day after L-cell inoculation or 3 weeks later, when tumors have grown to an appreciable size. The antitumor effect of poly(l)-ploy(C) was found to be dose dependent. Multiple injections of 100 microng poly(l)-poly(C) per mouse (congruent to 5 mg/kg) almost completely arrested tumor growth, but even at 1microng/mouse (congruent to 50 microng/kg), poly(l)-poly(C) caused a partial reduction in tumor growth. Mouse interferon administered exogenously in amounts that closely mimicked the interferon blood levels induced endogenously by poly(l)-poly(C) failed to duplicate the antitumor effects of poly(l)-poly(C). Hence, the role of interferon induction in the mechanism of antitumor action of poly(l)-poly(C) can be eliminated. Since all results were obtained with the use of athymic nude mice, the role of thymus-dependent immunity can also be eliminated. At the dosage regimens used to determine the inhibitory effect on L-cell tumor growth, neither poly(L)-poly(C) nor interferon altered the life-span of the nude mice.


Isolation of glycoprotein antigen from ascites fluid of pancreatic carcinoma. A glycoprotein antigen from the ascites fluid of pancreatic carcinoma has been isolated with the use of perchloric acid extraction and chromatographies on Sepharose 4B, DEAE-Sephadex, and DE52, followed by Sephadex G-200. The purified glycoprotein was found to be homogenous by sodium dodecyl sulfate polyacrylamide gel electrophoresis. A molecular weight of 185,000 dattons was determined by gel filtration. The molecule contained 55% protein and 45% carbohydrate. Both the amino acid and carbohydrate compositions were different from those of carcinoembryonic antigen. This glycoprotein antigen of pancreatic cancer cross-reacted with anticarcinoembryonic antigen; however, the antiserum prepared from this antigen did not react with carcinoembryonic antigen. The biological significance of this glycoprotein in pancreatic cancer is being studied.


Effects of antiinflammatory agents on mouse skin tumor promotion, epidermal DNA synthesis, phorbol ester-induced cellular proliferation, and production of plasminogen activator. The antinflammatory ateroids fluocinoine acetonide, fluocinonide, and fluclorolone acetonide were found to be very effectiveinhibitory agents of mouse skin tumor promotion. These steroids also drastically inhibited epidermal DNA synthesis and epidermal cellular proliferation induced by a phorbal ester tumor promoter. In addition, these compounds were potent inhibitors, of plasminogen activator production in tumor cell cultures. The clinically used non-steroidal antiinflammatory agents oxyphenbutazone, indomethacin, and Seclazone also inhibite tumor promotion but were much less effective. Although these agents are useful against inflammatory disorders in general when given p.o., in our studies they had little effect on inflammation and epidermal cellular proliferation induced by a phorbol ester tumor promoter when given topically. The afore mentioned nonsteroidal antiinflammatory agents also had little effect on epidermal DNA synthesis. Oxyphenbutazone and indomethacin were less potent inhibitors of plasminogen activator production in tumor cells than were the antiinflammatory steroids, and Seclazone produced a negligible inhibition. There is, therefore, a general correlation in the potencies of a series of steroidal antiinflammatory agents for inhibition of tumor promotion and their ability to inhibit plasminogen activator production by tumor cell cultures and epidermal DNA synthesis.


Mitochondrial inclusions in human cancer of the gastrointestinal tract. Four cases of adenocarcinoma of the gastrointestinal tract have been examined. Large electron-dense mitochondrial inclusions were found in the mitochondria of many cells; after detailed histochemical tests they could be classified into two types. The first type of inclusion consisted of clusters of electron-dense, calcium-containing granules linked to a glycoproteic substrate. These inclusions were linked to a glycoproteic substrate. These inclusions were always associated with cristae and were found in the mitochondria of cells that showed clear signs of degeneration. The second type of inclusion was found much more frequently and consisted essentially of phospholipids of which electron density was strictly osmium dependent. Their structure was usually at least partly lamellar, but in some cases it was homogeneous throughout. It is hypothesized that inclusions of the second type may have the same biological role as the morphologically identical inclusions found in the mitochondria of brown fatty tissue in the perinatal rat and in yeasts during glucose repression or anaerobiosis. The resemblance between the homogeneous variety of inclusions within the second type and the mitochondrial inclusions recently described in human leukemic lymphoblasts and monoblasts has been stressed to bring out the need for a histochemical check on the supposedly viral nature of the latter inclusions.


Effects of nine N-nitroso compounds on the specific radioactivity of liver proteins after injection of [14C]leucine into rats. We compared the effect of nine N-nitroso compounds, given by gavage to adult rats, on specific radioactivity of the trichloroacetic acid-precipitable liver proteins, 1 hr after the injection of [14C]leucine. The specific radioactivity was decreased by dimethylnitrosamine, diethylnitrosamine, methyl-n-butylnitrosamine, and nitrosomorpholine 5 to 10 hr after their administration; was increased by nitrosopiperidine, dinitrosopiperazine, and methylnitrosourea 5 to 24 hr after gavage; and was unaffected by nitrososarcosine and nitrosodihydrouracil. With dimethylnitrosamine, specific radioactivity was decreased by 10 but not 5 mg/kg. In control rats and rats given injections of either of two nitrosamines, protein specific radioactivity at 60 min after the [14C]leucine injection was 76 to 87% of that at 30 min, indicating some degradation of the proteins at 60 min. The liver:blood ratio of [14C]cycloleucine concentration was unaffected by four nitrosamines, indicating no effect on leucine transport. The effect of the nine compounds was examined on total pool size of free leucine in the liver, at times close to those for the maximum specific radioactivity effect. For these data, we calculated "corrected specific radioactivity," adjusted for changes in pool size. This adjustment is only a first approximation since, for example, the free leucine pool is not uniform with respect to protein synthesis. The four N-nitroso compounds that decreased specific radioactivity also decreased corrected specific radioactivity, even though they enlarged the leucine pool. Of the remaining compounds, two enlarged the leucine pool and three increased corrected specific radioactivity. For all nine compounds, the decrease in specific and correlated with the ability to cause acute liver necrosis. When nitrosodihydrouracil was excluded, the decrease in specific and corrected specific radioactivity was significantly correlated with the reported liver carcinogenicity.


Fasting and refeeding: cell kinetic response of jejunum, ileum and colon. Following a period of fasting, feeding a normal diet results in a burst of DNA synthesis in the crypts of the colonic epithelium. This is due largely to a prompt entry of cells, blocked in G1, into S. Peak levels of S cellularity exceed 4 times the fasting, and 2 times the normal fed control values. Refeeding a low residue diet (soluble casien, glucose and corn oil) results in a return to control levels of proliferative activity, but no hyperplasia. However, in jejunum and ileum, refeeding is followed by a return to near control levels of proliferation with only a slight overshoot in S phase cellularity. During the fasting period, the ileal crypt proliferative compartment (Pc-zone) and total crypt cellularity decline significantly. These changes are accompanied by an increase in the total cycle time, due to an equivalent lengthening of the G1 and S phase. Following refeeding, there is a reduction in the cycle time and a gradual return to the control values for the Pc-zone size and cellularity. In the colon, fasting has no effect on the Pc-zone size or total crypt cellularity. There is an approximate doubling of the cycle time due solely to an increase in G1. Following refeeding there is an increase in the Pc-zone size and crypt cellularity and a marked shortening of the cycle time. Evidence that a G1 cycle blockade is induced in the colon by fasting is given by a lenghening of the G1 period and by stathmokinetic studies employing vincristine.


Some properties of a "G0" -model of the cell cycle. II. Natural constraints on the theoretical model in exponential growth conditions. The two-phase (G and C phases) model first proposed by Burns & Tannock (1970) to describe the cell cycle kinetics has the major advantage of requiring only two parameters for a complete description of the kinetic behaviour of populations that are in a steady-state, or that grow exponentially (with no cell loss from the population). Steady-state populations were examined in paper I of this series. Exponential populations with no cell loss are investigated here. The model assumes two basic kinetic states--a 'C' phase which includes S, G2, M and perhaps part of G1, and a 'G' phase which cells enter after completing the C-phase and from which either are lost or return to C-phase randomly. The model assumes that transit time through C-phase is constant for all cells in the population. An original method is described which allows the determination of two independent parameters of the model from the experimental 'fraction of labelled mitoses' (FLM) curve; From those two parameters, the ratio of G-cells among the total number of cells (NG/N) has been calculated for each cell population studied, The range of the NG/N values thus obtained is fairly restricted, and the mean NG/N value for exponential growths is not statistically different from that found in steady-states considering in that case the only sub-population of cycling cells (i.e. the cells that will undergo a further mitosis).


A deficiency of hematopoietic stem cells in steel mice. In the present study, population sizes of high self-renewal potential stem cells, i.e. colony forming units (CFU), and low self-renewal potential stem cells, i.e. transient endogenous colony forming units (TE-CFU) in Sl/Sld mice and their normal congenic littermates were measured and compared. By correcting for differences in the seeding efficiency 'f', it was possible to demonstrate that Sl/Sld mice suffer a deficiency of both stem cell populations. Therefore, it is concluded that the defective stromal tissue of the Sl/Sld mouse does not support normal size stem cell populations. However, as noted in the discussion, it remains an open question as to whether the defective stromal tissue supports normal erythroid differentiation at the stem cell level.


The effect of vincristine on mouse jejunal crypt cells of differing cell age: double labelling autoradiographic studies using 3H- and 14C-TdR. The mechanism of action of the alkaloid vincristine (VCR) has been investigated in vitro on HeLa cells in culture and in vivo on jejunal crypt cells of the mouse. The in vitro experiments with HeLa cells show that VCR affects not only mitotic but also interphase cells. The VCR-affected cells first continue their passage through the cell cycle undisturbed but after reaching mitosis they are arrested in metaphase. This agrees well with the results obtained by Madoc-Jones & Mauro (1968) and Madoc-Jones (1973) on synchronized cell cultures. Until now there has been no investigation of the mechanism of action of VCR in vivo. This is due to the absence of a suitable technique for synchronization in vivo. The present study is based on a method which permits the assessment of the VCR sensitivity as a function of the cell age without synchronization in the usual sense. The jejunal crypt epithelium of the normal mouse was double labelled with 3H- and 14C-thymidine (TdR) in such a way as to produce a narrow subpopulation of crypt cells with a maximum age difference of 1 hr. On autoradiographs these cells can be distinguished by their characteristic labelling from other cells. As this 'pseudo'-synchronized subpopulation passes through the cycle the effect of VCR can be studied, i.e. one can analyse the effect in well-defined time intervals of the cycle. The results show that the effect of VCR is the same in vivo as in vitro. The crypt cells which are affected by VCR in interphase continue their passage through the cycle, but upon entering mitosis they are arrested in metaphase. VCR has, at the concentration used in the present study, no effect on the duration of the S and G2 phases. The necrotic cells seen after VCR application are formed from arrested metaphases.


The application of age distribution theory in the analysis of cytofluorimetric DNA histogram data. Age distribution theory has been employed in a model to analyse a variety of histograms of the DNA content of single cells in samples from experimental tumours growing in tissue culture. The method has produced satisfactory correspondence with the experimental data in which there was a wide variation in the proportions of cells in the intermitotic phases, and generally good agreement between the 3H-thymidine labelling index and the computed proportion in S phase. The model has the capacity to analyse data from populations which contain a proportion of non-cycling cells. However, it is concluded that reliable results for the growth fraction and also for the relative durations of the intermitotic phase times cannot be obtained for the data reported here from the DNA histograms alone. To obtain reliable estimates of the growth fraction the relative durations of the phase time must be known, and conversely, reliable estimates of the relative phase durations can only be obtained if the growth fraction is known.


Cytokinetics of rat tracheal epithelium stimulated by mechanical trauma. Mild abrasion of rat tracheal epithelium results in irreversible damage to the superficial cells and stimulates the viable basal cells to participate in a nearly synchronous wave of DNA synthesis and mitosis. For the growth population as a whole, DNA synthesis started at 14 hr after injury and persisted for 16 hr. The duration of S in individual cells was determined autoradiographically by identifying the time at which a second pulse of DNA precursor (14C-TdR) was no longer incorporated by cells labelled with 3H-TdR at the onset of S. S was found to be 8-9 hr long. It was also determined that cells entering S at later times synthesized DNA for the same 8-9 hr period. TG2 was calculated to be 2 1/2-3 1/2 hr by subtraction of TS and 1/2TM from the period from onset of DNA synthesis to metaphase. By making a second denuding lesion adjacent to the first injury, the cells were stimulated through at least another period of S. At the peak of the second wave of DNA synthesis (50 hr after injury) 14C-TdR was present in the same cells which had incorporated 3H-TdR administered at the mid-point of the preceeding synthetic phase. The 28-hr interval between these two peaks of synthesis is the measure of cell cycle duration for these regenerating tracheal epithelial cells.


Lectin binding to dissociated cells from two species of Xenopus embryos. 1. Fluorescein isothiocyanate-conjugated concanavalin A (F-conA) and soy bean agglutinin (F-SBA) bind to the surface of EDTA-dissociated cells from blastula and gastrula stage Xenopus laevis and X. mulleri embryos. 2. Binding of these lectins is abolished by appropriate haptens (alpha-methyl-D-mannopyranoside for F-conA and 2-acetamido-2-deoxy-D-galactose for F-sba). 3. Gastrula stage cells show a clustering or capping of lectin binding sites not shown by blastula stage cells. 4. At least for F-conA, this capping is induced by the lectin. 5. There are no striking regional differences in either amount or pattern of lectin binding in early gastrulae of both species.


Studies on the structure and possible function of the RNA "cap" in developing sea urchins. Paracentrotus lividus embryos at the hatching blastula stage very quickly incorporate radioactivity from labeled nucleosides (except uridine) or 32P- or methyl-labeled methionine into a portion of the RNA that has been identified as a "cap". The most probable sequence of this cap is m7G (5') ppp (5')mAmpCp. A very active "capping" and methylation of the "cap" of preexisting RNA molecules was shown to occur at the blastula stage.


RNA synthesis in spermatocytes and spermatids and preservation of meiotic RNA during spermiogenesis in the mouse. The rate of RNA synthesis at different stages of spermatogenesis in the mouse, and the preservation of RNA from the diploid to the haploid phase of spermatogenesis were studied in homogeneous germ cell fractions separated by velocity sedimentation at unit gravity. The uridine pool expansion method was used for determining the rate of RNA synthesis: seminiferous tubules were labelled in culture with increasing concentrations of [3H]-uridine and the incorporated radioactivity was estimated in cell fractions separated by velocity sedimentation. The results indicate that before nuclear elongation, round spermatids (steps 1 to 8 of spermiogenesis) synthesize RNA at the same rate per DNA content as middle-late pachytene spermatocytes. The preservation of RNA molecules synthesized in meiosis was investigated by labelling pachytene spermatocytes with T3H]uridine in vivo and collecting samples of germ cells at definite stages of spermatogenesis at various time intervals thereafter. The results show that a considerable proportion the RNA synthesized during the pachytene stage is preserved through spermatid development until late spermiogenesis.


Analogs of natural lipids. II. Polymorphic behavior of the tris-homoacyl derivatives of cyclopentane-1,2,3-triols. The polymorphic behavior of three series of tris-homoacyl (C14:0, C16:0 and C18:0) cyclopentane-1,2,3-triol analogs of the natural saturated triglycerides has been studied using differential thermal analysis, Fourier transform infrared spectroscopy, and X-ray diffraction. It was found that the triglyceride analogs derived from the 1,2,3/0 and 1,2/3 cyclopentanetriols exhibit different polymorphic behavior than that of the natural triglycerides. The analogs derived from 1,3/2 cyclopentanetriol, however, were found to parallel the polymorphic behavior of the natural triglycerides quite closely. This polymorphic behavior is discussed in terms of the different configurations which the chains assume in each of the triglyceride analogs.


Effects of stroke volume and velocity of ejection on end-systolic pressure of canine left ventricle. End-systolic volume clamping. To study the effects of contraction mode on ventricular end-systolic pressure-volume relationship, we compared the end-systolic pressure of isovolumic contraction with that of ejecting contraction at an identical end-systolic volume. The left ventricle of excised cross-circulated canine hearts was fitted with a water-filled balloon. The balloon was connected to a hydraulic pump that allowed the ventricle to contract to a preset constant end-systolic volume (19-37 ml) from a variable end-diastolic volume. At each of control, enhanced, and depressed levels of contractility, differences of end-systolic pressures of steady state isovolumic and ejecting contractions were evaluated while stroke volume and velocity of ejection were widely varied. The end-systolic pressure in the ejecting contraction tended to decrease by 5-15% from that of the isovolumic beat with increases in either stroke volume to 20-25 ml or peak velocity of ejection to about 800 ml/sec. There was no obvious difference in the results at different levels of contractility. The magnitude of the end-systolic pressure depression due to ejection was, however, relatively small as compared to 4-fold changes in end-systolic pressure due to the changes in contracility. We, therefore, conclude that the ventricular end-systolic pressure-volume relationship is affected slightly by ejection, and that this effect is much smaller than the maximal effect of changing contractility on the end-systolic pressure-volume relationship.


Input impedance of the systemic circulation in man. To determine the systemic input impedance, pulsatile pressure and flow were measured in the ascending aorta in 16 human subjects who were undergoing diagnostic cardiac catheterization. Blood flow was measured with a catheter-tip electromagnetic velocity meter, and pressure with an external transducer connected with the fluid-filled lumen of the catheter. Five subjects were found to have no evidence of cardiovascular disease (group A, mean age 32 +/- 2 years, mean aortic pressure 97 +/- 4 mm Hg). Seven had clinical and angiographic signs of coronary arterial disease, and mean pressures less than 100 mm Hg (group B, mean age 48 +/- 2 years). Four subjects had signs of coronary disease and mean pressures greater than 100 mm Hg (group C, mean age 48 +/- 3 years). The frequency spectra of impedance were qualitatively similar in all three groups and resembled those previously observed in the canine aorta. Characteristic impedance was lower in the normal subjects (group A, average 53 dyn sec cm-5) than in the subjects with coronary artery disease (groups B and C, average 129 dyn sec cm-5). Among the subjects with coronary disease, characteristic impedance was higher in the hypertensive subjects (group C, average 202 dyn sec cm-5) than in those with lower mean pressures (group B, average 95 dyn sec cm-5). External left ventricular work per unit time (hydraulic power) averaged 1715 milliwatts (mW) in group A, 1120 mW in group B, and 2372 mW in group C. Cardiac outputs were within normal limits in all subjects, but tended to be lower in group B than in group C. These results suggest that the subjects of group C were better able to meet the increased energy demands imposed by an abnormally high aortic input impedance. Further investigation is needed to learn whether the high impedances in subjects with coronary disease represent an increase with age and transmural pressure alone, or whether some additional factor is involved. The data on relatively normal subjects permit a tentative definition of the normal limits for aortic input impedance in man: 26-80 dyn sec cm-5.


Differences between proximal left and right bundle branch block action potential durations and refractoriness in the dog heart. To date the electrophysiological mechanism responsible for aberrant intraventricular conduction of critically timed premature supraventricular impulses has not been documented. Microelectrode techniques were used to measure in vitro action potential and refractory period durations of the canine proximal right and left bundle branches equidistant from the distal bundle of His. Both measurements in the right bundle branch were statistically significantly longer than these parameters of the left bundle branch. Transection of the bundle branches immediately distal to the distalmost recording sites effected no change in the proximal right bundle action potential but caused marked prolongation of proximal left bundle branch action potential and refractory period durations. We conclude that functional right bundle branch aberrancy is most likely due to the longer proximal right bundle action potential duration and refractoriness. Our data also suggest that the shorter proximal left bundle branch action potential durations and refractory periods may be due to the proximity of the low ohmic resistance Purkinje fiber-muscle junctions on the left septal surface, effecting electrotonic foreshortening of these proximal left bundle branch parameters.


Effects of the antibiotic ionophore X-537A on contractility and ionic exchange in rabbit ventricular myocardium. The effects of the ionophore X-537A on mechanical function and on ionic exchange were studied in the isolated, arterially perfused rabbit interventricular septum. X-537A produced an initial positive inotropic response which was, however, transient in this preparation and appeared to be dependent on an effect of the ionophore on catecholamines. The positive inotropy gave way to a progressive decline in force development which was unrelated to the action of catecholamines and was not accompanied by the development of contracture. Isotope uptake experiments revealed that coincident with this decline in force development there was a continuous net loss of tissue K+ and a net gain of Na+. X-537A (5 micronM) perfused for 20 minutes resulted in a net K+ loss of 50.2 +/- 4.6 mmol/kg dry weight and a net Na+ gain of 74.0 +/- 4.5 mmol/kg dry weight. Isotope washout experiments confirmed that the entire net loss of K+ could be accounted for by increased K+ efflux. X-537A did not alter Na+ efflux nor did it have any detectable effect on 45Ca exchange of the perfused septum in which the ability to detect net movements is at a level of approximately +/- 550 micron mol/kg dry weight. On removal of the ionophore the decline in force development ceased and reversed to near control levels and the progressive ionic changes ceased. However, despite the near total recovery of contractile function the Na+ and K+ levels remained at values little different from those reached at the termination of X-537A perfusion. In addition, after removal of the ionophore, we found that K+ exchange rate remained significantly less than control and, furthermore, a kinetic inhomogeneity of tissue K+ was observed. The results emphasize a dissociation between cellular Na+ and K+ content and function following the ionic perturbations induced by the ionophore.


Effect of hematocrit on the washout of xenon and iodantipyrine from dog myocardium. The rates of washout of 133Xe and 131I-iodoantipyrine from the myocardium into the coronary sinus blood were determined over a wide range of hematocrits after simultaneous injection of the isotopes into the left anterior descending coronary artery of dogs. The ratio of the monoexponential disappearance constants (kX/kI), which is a measure of the ratio of the dynamic blood-tissue partition coefficients (lambdabIX/lambdabtI), increases linearly with hematocrit. This dynamic lambdabt ratio has almost the same relationship to hematocrit as the static lambdabt ratio. By the use of appropriate hemotocrit-specific lambdabt values, we found that simultaneous blood flows calculated from the two indicators with different disappearance constants showed excellent agreement. If this hematocrit-dependent alteration in indicator partitioning is neglected, blood flow measurement in the dog with 133Xe washout may introduce an error of approximately 1.15% per unit hematocrit deviation from the normal value (taken as 45%), whereas blood flow measurements with 131I-iodanatipyrine washout had negligible errors.


High-performance liquid chromatography of pyruvic and alpha-ketoglutaric acids and its application to urine samples. Pyruvic and alpha-ketoglutaric acids were quantitatively and rapidly converted to quinoxalones by a new method, to enhance their detectability in the ultraviolet. The quinoxalones were separated by reversed-phase chromatography with use of 10 micronm particles packed in a 30-cm (length) column. An aqueous solution of ammonium acetate and methanol was used for linear gradient elution. The urinary excretion of pyruvic and alpha-ketoglutaric acids by normal individuals, diabetic patients, and patients with renal dysfunction was determined.


Some problems associated with assay of 25-hydroxycalciferol in human serum. Methods available for the assay of 25-hydroxycalciferol in human serum are evaluated and compared to one another. Ethanol was chosen for use in the initial extraction procedure and rat-kidney cytosol as the binding protein, although good alternative methods are also available. We used silicic acid for chromatography and found this an essential step. Reproducibility was increased when, after "bound" and "free" material were separated, an aliquot of the supernate was pipetted into the counting vial instead of the entire supernatant fluid being decanted. Beta-lipo-protein added to the assay system was of no advantage; added bovine serum albumin interfered with the assay by giving rise to high blank values. With ethanol extraction, silicic acid chromatography, rat kidney cytosol and separation on dextran-coated charcoal, sera from normal subjects showed a mean 25-hydroxycalciferol concentration of 28.5 microng/liter (range, 13.1 to 43.9) during the fall season. Coefficients of variation for a control serum were 4.9% (intra-assay) and 10.9% (interassay).


Stability of commonly used thiols and of human creatine kinase isoenzymes during storage at various temperature in various media. We assessed the stability--at 20, 4 --20, and --80 degrees C--of 2-mercaptoethanol, dithioerythritol, and dithiothreitol, and of semi-purified creatine kinase isoenzymes of human origin in the presence and absence of the three compounds. The isoenzymes plus the sulfhydryl compounds were also assessed an aqueous buffer, in heat-inactivated pooled sera, or in fresh pooled sera with low endogenous total creatine kinase. We also examined the influence of thawing conditions from--20 or--80 degrees C, of fast freezing, and of exposure to fluorescent light. At 20 and 4 degrees C some--SH groups are oxidized. At--20 and--80 degrees C this loss is diminished. It is a function of both temperature and diluent. For stability, creative kinase isoenzymes stored at any temperature require the presence of a suitable sulfhydryl compound. Periodic addition of fresh sulfhydryl solutions stabilized creatine kinase isoenzymes suspended in the two protein-based diluents by 10-30%; for those in an aqueous buffer such additions decreased the activity to stored creatine kinase isoenzymes. Fast freezing does not increase recovery of creatine kinase isoenzyme activity over slow freezing at--80 or--20 degrees C. Thawing should be done as infrequently as possible and should be done rapidly, at 37 degrees C, for the residual enzyme activity to be maximal.


Intra-individual variation of some serum constituents and its relevance to population-based reference ranges. The advent of high-capacity multi-channel analyzers allows estimation of long-term variability in serum constituents of large numbers of subjects. By frozen storage of specimens with subsequent analysis in a single machine run, long-term analytical variation may be eliminated, thus sharpening the estimates of intra-individual variation. In the present study we used the Vickers M-300 analyzer to obtain the data for such estimates from 37 male volunteers, each bled once a week for 22 weeks. Secimens were analyzed in random order to eliminate any biasing effect of analytical drift during the 4-h machine run. Ten serum constituents were measured. Storage-induced linear trends were small or negligible during the period of specimen collection. Using the ratio of average within-subject variance to the variance among subjects as a guide, serum alkaline phosphatase was found to show the greatest individuality, sodium and potassium the least. Other constitutents showed varying degrees of individuality, but for all these analytes, the usual population-based reference ranges were found to be either insensitive or irrelevant to the study of concentration changes over time within most healthy subjects. Our results generally confirmed those of smaller but comparable earlier studies.


Gas-chromatographic profiling of urinary acidic and alcoholic catecholamine metabolites. Urinary acidic and alcoholic catecholamine metabolites were isolated separately from 24-h urines by simple extraction procedures. After derivatization, extracts were analyzed by means of simultaneous gas chromatography on two different stationary phases. The quality of the assay was checked by running "normal" and enriched commercial (Hyland) control urines in each series. Normal values are given for vanilmandelic acid, homovanillic acid, and 3-methoxy-4-hydroxyphenylethylene glycol, expressed as a function of age. We present excretion patterns showing these metabolites and also 3,4-dihydroxyphenylacetic acid, vanillacetic acid, 3,4-dihydroxyphenylethylene glycol, and vanilethanol for 12 patients with neurogenic tumors. Values obtained for vanilmandelic acid by this method correlate well with those obtained by the electrophoretic/colorimetric method of Hermann [Am. J. Clin. Pathol. 41, 373 (1964)].


Myoglobin concentrations and muscle-enzyme activities in serum after myocardial infarction and cardiac arrhythmia. Sera from patients with myocardial infarction and cardiac arrhythmias were analyzed for myoglobin concentration and the activities of total creatine kinase, creatine kinase isoenzyme-2, and lactate dehydrogenase isoenzyme-1 at the time of hospital admission and during the first few days of hospitalization. The nine patients with a final diagnosis of myocardial infarction had abnormally high values for total creatine kinase, creatine kinase-2, lactate dehydrogenase-1, and myoglobin. Myoglobin concentrations were highest on admission in six patients and on the day after admission in the other three patients. Creatine kinase-2 manifested maximum activity on the day after admission for all patients with myocardial infarction. Lactate dehydrogenase-1 did not reach maximal values until the second or third day after admission. The six patients with arrhythmias did not show any significant increases in creatine kinase-2 or lactate dehydrogenase-1. Myoglobin and total creatine kinase, however, were increased in the four patients who had received cardioversion. The specificity and diagnostic usefulness of these serum measurements are discussed.


Comparison of several methods for semiquantitative determination of urinary protein. Two types of urine protein dipsticks and the sulfosalicylic acid method were compared for their accuracy and specificity, with use of urine samples supplemented with various proteins. Dipsticks yield accurate results when the protein under consideration is restricted to albumin; the sulfosalicylic acid method accurately determines many kinds of proteins in addition to albumin. Detergents affect each of the methods, but changes in salt concentration only affect results by dipstick procedures. Dipsticks, which are based on the protein-error principle for indicators, are subject to some of the conditions that apply to the bromcresol green method for serum albumin determination.


A new colorimetric method for the determination of plasma lecithin-cholesterol acyltransferase activity. A new method has been developed to improve the linearity of the plasma lecithin-cholesterol acyltransferase reaction by adding synthetic dipalmitoyl lecithin sol to the incubation medium and subsequently measuring the change in free cholesterol content after incubation by a combined enzymatic method using cholesterol oxidase and peroxidase. The method was found to be simpler than other techniques such as gas-liquid chromatography or isotope methods and therefore may be suitable for the routine assays of plasma lecithin-cholesterol acyltransferase activities in clinical laboratories.


A new colorimetric method for determination of serum glucose. A new colorimetric method using 4-aminoantipyrine-dimethylaniline in a glucose oxidase-peroxidase system is proposed to determine glucose in serum. The developed color was stable at the optimum pH of glucose oxidase, 5.6. The method is more sensitive than the o-dianisidine method and is reproducible. The correlation between the proposed method and the o-dianisidine method is 0.989.


A gas chromatographic method for the estimation of phenprocoumon, 3-(1-phenyl-propyl)-4-hydroxycoumarin (Marcoumar, Liquamar), in human serum or plasma. A rapid and reliable gas-chromatographic method is described for the estimation of phenprocoumon from human serum or plasma. No interference was found from other usual oral anticoagulant drugs. The method can be used for therapeutic and toxic levels. The appearance of two volatile derivatives of phenprocoumon (in constant ratio) is discussed.


Inhibition of human placental-type alkaline phosphatase variants by peptides containing L-leucine. The inhibition of normal human placental alkaline phosphatase, its rare "D-variant" phenotype, and the "Nagao isoenzyme" found in an ovarian cancer fluid by peptides containing L-leucine were studied. The Nagao isoenzyme and the D-variant phenotype were distinct from the normal placental enzyme, but similar to each other in their inhibition by L-leucine and dipeptides containing L-leucine in their amino termini. On the other hand, L-leucinamide, L-Leu-L-Leu-L-Leu and L-Leu-Gly-Gly inhibitions did not readily discriminate between D-variant and normal phenotypes, but did so between D-variant and Nagao isoenzymes. The distinctions among these enzymes in their responses to peptide inhibitors may reflect differences in their primary structures.


A rapid column chromatographic procedure for the routine measurement of taurine in plasma of normals and depressed patients. A rapid method has been developed for the routine measurement of taurine in plasma. A preliminary clean-up step has made it possible to assay multiple samples in a relatively short period of time using the excellent sensitivity and specificity of the analytical column of the automatic amino acid analyzer. The speed and specificity of the method permits its use for the routine measurement of taurine in blood samples. Employing this method, a value of 45+/-4nmol taurine per ml platelet free plasma (mean+/-S.E.M.) was obtained for normal healthy adults. Analysis of plasma from depressed patients who are free of drugs as well as from depressed patients under treatment with drugs gave values of 29+/-2 and 25+/-3 nmol taurine per ml platelet free plasma (mean+/-S.E.M.), respectively.


Minimal effects of free fatty acids on the competitive protein-binding assay of serum thyroxine on reusable Sephadex columns. In rats; following oral administration of cream and intravenous heparin, plasma free fatty acids and triiodothyronine Sephadex uptake were elevated. Simultaneously apparent serum thyroxine, measured by the Tetrasorb-E kit, was increased, but not when measured using reusable Sephadex columns. Addition of oleate to a human serum albumin solution or to human serum produced similar effects; appreciable rises in apparent thyroxine were noted in the kit assay but only minor changes occurred when Sephadex columns were used. In the latter procedure almost all [14C]oleic acid added to serum was removed in the first barbital buffer wash, before application of the thyroxine-binding globulin solution. In the serum thyroxine assay on reusable Sephadex columns, there is minimal interference by free fatty acids because they are removed by the first barbital buffer wash.


Immunochemical heterogeneity of calcitonin in man: effect on radioimmunoassay. Determinations of blood levels of human calcitonin by radioimmunoassay have varied considerably in different laboratories. Much of the controversy over calcitonin levels can be attributed to the multiplicity of immunoreactive forms of the hormone (iCT), the differing region specificities of the antisera utilized for measurement by radioimmunoassay, protein effects, different rates of degradation of the various iCT fractions and the specific methodology of the radioimmunoassay.


Quality control in the simultaneous assay of anticonvulsants using an automated gas chromatographic system with a nitrogen detector. An automated gas-liquid chromatographic technique for the routine determination of phenobarbitone, primidone and phenytoin using a nitrogen-detector is described. The high selectivity of the detector for nitrogen-containing compounds allows a direct extraction procedure and the use of a phenytoin analogue as an internal standard obviates the need for accurate aliquot measurement after extraction. The anticonvulsants are chromatographed as methyl derivatives produced by on-column methylation with trimethylphenyl ammonium hydroxide following automatic liquid injection. Quality control data from an internal quality control scheme is presented and the performance of the laboratory in an inter-laboratory control scheme is reported and discussed.


Heterogeneity in electrophoretic mobility and in molecular size of lactate dehydrogenase-immunoglobulin G complexes. Some human sera have been found to contain lactate dehydrogenase (LDH)-immunoglobulin complexes. A serum with LDH-IgG3 complexes was further analysed. Complexes with different net charge and different molecular size were found. Their isoelectric points, measured by isoelectric focusing, were in the range of pH 6.3 to 7.2. Gel-filtration experiments showed that the molecular size varied from about 300 000 to 1 000 000. Most of the complexes could be precipitated from solution with 30--40% saturated ammonium sulfate or a low ionic strength.


Estimation of hydroxylysine in urine and serum of patients with chronic uremia. Free hydroxylysine and hydroxylysing glycosides were separated from urine and serum extracts on cation exchange resin and assayed spectrophotometrically. The method in conjunction with gel filtration in Bio-Gel P2 allowed to separate from urine also polypeptide hydroxylysine and hydroxylysine bound in small molecules of neutral or acidic character. Glycosylgalactosylhydroxylysineand galactosylhydroxylysine were separated by partition and/or ion exchange chromatography. Patients with chronic renal insufficiency had elevated serum levels and urinary excretion of hydroxylysine glycosides with increased excretion of hydroxylysine bound in polypeptides and in small molecules of neutral or acidic character. The excretion of free hydroxylysine was often within normal limits. When compared to values found in normal growing subjects and in adult patients with increased bone turnover and normal renin function the urinary excretion of hydroxylysine glycosides in chronic uremia was more markedly increased than excretion of hydroxyproline polypeptides and total hydroxyproline.


Preparation of high-quality iodine-125-labelled pituitary human follicle-stimulating hormone (hFSH) for radioimmunoassay: comparison of enzymatic and chloramine-T iodination. A method is described for the enzymatic radioiodination of human follicle-stimulating hormone (hFSH) by a system consisting of lactoperoxidase, hydrogen peroxide and Na125I. It was compared with the Chloramine-T modified technique. A satisfactory specific activity of the labelled hormone was obtained with the enzymatic iodination with much greater immunoreactivity was stability than after Chloramine-T.


Age-related changes in human skin collagen galactosyltransferase and collagen glucosyltransferase activities. Collagen galactosyltransferase and collagen glucosyltransferase activities were assayed in human skin specimens of about 100 mg wet weight. The assay of the glucosyltransferase activity was found to be highly specific. The assay of the galactosyltransferase activity was somewhat less specific, but there was no difference in specificity between the foetal and adult human skin samples. The activities of the two collagen glycosyltransferases in human skin extract were found to vary with age, being highest in foetal skin, and higher in the skin of young children that in that of adults. The galactosyltransferase and glucosyltransferase activities in foetal skin were respectively about 4 times and 6 times those in adult skin. The magnitudes of the changes with age in the two collagen glycosyltransferase activities were smaller than those occurring in the activities of the two other intracellular enzymes of collagen biosynthesis namely prolyl and lysyl hydroxylase. This difference suggests that the four intracellular enzyme activities of collagen biosynthesis are not regulated in an identical manner.


Serum triiodothyronine: clinical experience with a new radioimmunoassay kit. A new kit for measuring total serum triiodothyronine (T3) by radioimmunoassay ('T3 RIA'; Radiochemical Centre, Amersham) was evaluated using sera from 1114  patients and normal controls. The kit performed reliably with intraassay and interassay variability figures of 3.9% and 9.3%, respectively, at 'medium' concentrations of T3. A T3 measurement could be obtained conveniently within 18 h after overnight incubation at room temperature. There were no critical steps dependent on time or temperature. Serum T3 values showed no significant sex difference. There was no significant change in mean serum T3 between the ages of 15 and 69 years, but it fell by 0.15 nmol/l for every 5 years beyond the age of 70. Mean serum T3 (+/-SD) for 335 normal euthyroid subjects ages 15-69 years was 2.11+/-0.46 nmol/l (range: mean+/-2 SD = 1.19-3.03 nmol/l). 64% of pregnant euthyroid women and 12% of those taking oral contraceptives had elevated serum T3 levels, as did all hyperthyroid patients, apart from one with T4 toxicosis. Overlapping T3 values from hyperthyroid patients and euthyroid subjects with elevated concentrations of thyroid binding proteins could be separated completely by two correction techniques which related total serum T3 to the corresponding T3 resin uptake test, viz an 'augmented free T3 index', or a map plot of T3 vs T3 resin uptake. A 25% incidence of T3 toxicosis was observed. One hyperthyroid patient with T4 toxicosis, and seven euthyroid patients with 'biochemical T4 toxicosis' were investigated. 73% of moderately and 30% of severely hypothyroid patients had normal serum T3 levels. This overlap was not reduced by applying correction techniques. Our studies demonstrate the value of serum T3 measurements in screening  for and diagnosing hyperthyroidism. As T3 measurements become more widely available, it would seem reasonable to subclassify hyperthyroid patients into three types: those with 'T3/4 toxicosis', 'T3 toxicosis' and 'T4 toxicosis'.


Synchronous nyctohemeral rhythms in human blood melatonin and in human post-mortem pineal enzyme. Serum melatonin was measured by radioimmunoassay at 4-hourly intervals for 24 h in five normal men, and shown to exhibit nyctohemeral variations. The highest concentrations of serum melatonin occurred at 02.00 hours and the lowest concentrations at 14.00 hours. The synthetic enzymes for melatonin, serotonin-N-acetyl transferase (SNAT) and hydroxyindole-O-methyltransferase (HIOMT) were measured in post-mortem pineal organs from thirty-six men and women, and the activity of both enzymes was directly related to the time of death of the subject, resulting in a similar nyctohemeral curve as that observed with serum melatonin. The SNAT and HIOMT enzymes were stable when measured in pineal organs held under the usual post-mortem conditions. The observations indicated a nyctohemeral rhythm in human pineal function and suggests that any clinical interpretation of pineal dysfunction in disease states should take into consideration this normal rhythm.


Chromosome polymorphisms in karyotypes from amniotic fluid cell cultures. Frequencies of 12 fluorescent chromosome polymorphisms were scored from Q-banded karyotypes derived from 108 midtrimester diagnostic amniotic fluid cell cultures. The most frequent variants were the bright fluorescent short arm of chromosome 13 (P = 0.458) and the bright fluorescent marker close to the centromere on chromosome 3 (P = 0.426).The polymorphism pattern was found to be different between the maternal (blood culture) and fetal (amniotic fluid cell culture) karyotypes in each of the 25 paired cases studied. This technique is valuable in prenatal diagnosis to exclude possible maternal cell contamination and outgrowth in cases where the amniotic fluid cell cultures reveal a female karyotype.


Apparently balanced de novo translocations in patients with abnormal phenotypes: report of 6 cases. Six patients have been ascertained because of abnormal phenotypes but with apparently balanced de novo translocations. Five of them were mentally retarded with multiple congenital anomalies. The sixth patient had normal mental development but revealed ambiguous genitalia and multiple congenital anomalies. No syndromal diagnosis was possible in any of the six cases. The appearance of apparently balanced reciprocal translocations in association with abnormal phenotype may be coincidental, or the two may be causally related. If the latter is true, the causal relationship may be based upon: 1) a submicroscopic chromosomal loss, 2) position effect, or 3) a mutation at the site of the break in one or both translocated chromosomes.


The incidence of Duchenne muscular dystrophy in the South East of Scotland. In a survey carried out to determine the incidence of Duchenne muscular dystrophy in the South East of Scotland, 47 cases were ascertained among 177,413 live male births for the years 1953 to 1968. The overall incidence is 1 case in 3,775 live male births (26.5 +/- 3.2 X 10(-5)). This incidence figure has been compared with those reported in nine other studies, and any differences appear likely to be due to varying degrees of ascertainment. So far there is no evidence of any decrease in incidence of the disease in S.E. Scotland as a result of genetic counselling and antenatal foetal sexing.


The acrocephalosyndactyly syndromes: a metacarpophalangeal pattern profile analysis. Metacarpophalangeal pattern profile (MCPP) analysis was applied to patients with Pfeiffer and Chotzen syndromes, dominantly inherited types of acrocephalosyndactyly (ACS). A characteristic MCPP was obtained for the group. However, it did not discriminate between patients with Chotzen syndrome and those with Pfeiffer syndrome. Six patients in a single family showing Pfeiffer syndrome exhibited this unique MCPP profile which was not present in non-affected family members. Furthermore, three normal-appearing relatives were identified as affected by this technique, and this diagnosis was subsequently confirmed by radiographic and genetic data. The results show that: 1) the MCPP in Pfeiffer and Chotzen syndromes is unique and can be used to make the diagnosis of an ACS syndrome in persons who show few or no clinical stigmata of disease; 2) the MCPP does not discriminate between these two ACS types.


Knee ligament injuries: a standardized evaluation method. An analysis of the data with total scores in non-acute terms obtained from 280 patients suggests that knee injuries can be evaluated as: Good-Excellent=generally normal or near normal function, with few or no signs and minimal symptomatology, 41-50 points; Fair (+)=slight functional disability, with few signs and mild symptomatology, 36-40 points; Fair (-)=moderate functional disability, with significant clinical signs and moderate symptomatology, 31-35 points; Poor=severely disabled, with marked signs and symptoms of a compromised knee, less than 30 points. These gross categorizations correlate well with the patient's own evaluations of his condition and the physicians' clinical assessment of the status of a given knee. Although the majority of patients have been followed for four years or less with this method, it is felt that the method presented has provided: A consistency and organization of the workup, diagnosis, treatment, and study of knee ligament injuries, not previously available at our institution; an objective measure of individual patient response to treatment and eventual recovery; in general, more objective means of assessing modalities of therapy and modes of primary repair and reconstruction of ligaments; a tool facilitating communication both among physicians and between physicians and patient; a means by which the long term follow-up of an individual patient by more than one physician can become a more efficient undertaking; an increasing data bank of standardized observations from which many ideas are yet to evolve; and most importantly, an improvement and the means possible for further improvement in the care rendered patients with knee ligament injuries; also, the present computerized format has given rise to additional benefits not directly related to the immediate problem of knee ligament injury. One such benefit is the ability to request and receive rapidly, a concise, yet complete legal summary of a patient's hospital course.


Talipes equinovarus: "hard" versus "soft" tissues. Nonsurgical treatment is contraindicated in clubfeet which are obviously resistant. Early surgical release of the contracted "soft" tissues can effectively prevent the above described "hard" tissue deformities.


The natural course of untreated osteoarthritis of the knee. Ninety-four knee joints in 71 patients had primary gonarthrosis (osteoarthritis of the knee) for periods of 10-18 years. The course of gonarthrosis was unfavorable in that a majority of the cases became worse over the years and some also developed pain at rest. The prognosis in gonarthrosis seemed worse than in coxarthrosis (osteoarthritis of the hip). The radiographic changes in the affected knee joint progressed over the years. However, in most knees the changes remained limited to the compartment first affected so that knees with initially medial involvement had exclusively medial involvement even 10-18 years later. Varus deformity was related to poor stability and to a generally poor prognosis. An early onset of symptoms was unfavorable with regard to development of pain. The majority of otherwise healthy patients became incapable of using public transportation because of knee pain on walking.


Resurfacing of femoral head after osteotomy: a report of three cases. Three cases of osteotomy for osteoarthritis of the hip which demonstrate the ability of cartilage to resurface previously eburnated areas are presented. These cases support the claim that cartilage regeneration of the femoral head after osteotomy does take place. Histologically, this new surface consists of fibrocartilage.


The ultrastructure of the osteoclast and its functional implications. Recent findings on the ultrastructure of the osteoclast indicate that special attention should be given to the ruffled border, clear zone, and the vacuoles and vesicles of the cell and their significance for the mechanism of breakdown of bone matrix. The ruffled border is seen as an extensive area of cell surface where secretion of enzymes as well as uptake of matrix components takes place. The clear zone encircles the ruffled border completely and thus forms an integral part of the resorbing apparatus. Vacuoles and vesicles are thought to secret enzymes as well as take up extracellular material and possible digest or transport these products in the cell. The changes that occur in the ultrastructure of the osteoclast after exposure to parathyroid hormone and calcitonin indicate an important role of the ostioclast in bone metabolism. The cell can increase its activity very rapidly in response to parathyroid hormone, and decrease its activity in response to calcitonin.


Bone strength: before and after removal of unthreaded and threaded pin and screws. Bones with retained screws gradually regain normal strength but upon removal the strength drops to that of bones with fresh holes. This strength loss is probably attributable to microdamage occurring to the bone surfaces at the time of removal.


The reversal of tissue differentiation around screws. Mechanical factors influence the morphology and function of differentiated as well as undifferentiated cells. Observations of tissues surrounding screws, during a 4-week period of stability followed by a 4-week period of instability, demonstrate resorption of the newly formed callus and subsequently to screw lossening. Four weeks of instability followed by 4 weeks of stability produced not only a cessation of the osteoclastic activity, but also the appearance of an osteoclastic activity. Thus, the holding power of screws is not finally determined at the moment of their insertion nor after cellular differentiation has occurred around the screws. Local mechanical conditions throughout the period of fracture consolidation are of eqaul importance.


Epiphyseal tilt produced by experimental overload. After segmental resection of the diaphysis of the radius in growing dogs, the distal ulna epiphyseal plate is submitted to a mechanical overload which provokes a progressive tilt of the epiphysis. A full exail realignment is developed in two or three months as shown in radiographic follow-up studies. Histologically, the first lesion is a fissure starting and the convex side of the growth line, passing along the epiphyseal plate and located somewhere between the proliferative layer which is seldom affected and the zone of enchondral ossification. The separated segment of the growth cartilage situated between the fissure and metaphysis stops growing and the cartilaginous tissue becomes disorganized and necrotic. It is then reabsorbed, and replaced by vascular-connective tissue coming from the bone marrow.  Growth usually goes on form the deep proliferative layer of the epiphyseal plate when its vascular connections are intact. After three months a normal epiphyseal line is rebuilt correcting the defective axis of the bone.


Postmenopausal osteoporosis: cause, prevention and treatment. At this point, I think I should point out that it now seems possible that postmenopausal osteoporosis, like smallpox and poliomyelitis, can be eradicated. Although full replacement doses of oestrogen prevents progression of the disease in women who have already lost a great deal of bone, this is like locking the stable door after the horse is gone. Present evidence is that smaller doses will probably suffice to prevent loss of bone at the menopause: mestranol 20 microng (Aitken et al, 1973) or conjugated oestrogens USP 0.625 mg (Meema, Bunker and Meema, 1975). The minimal prophylactic osteotrophic dose has not yet been determined since the two doses indicated were the smallest tested. These two studies provide hope that a dose can be found which will prevent postmenopausal osteoporosis without necessarily producing endometrial hyperplasia. It is very likely that the prevention of bone loss by small, prophylactic doses of oestrogen will reduce the number of fractures of vertebrae, wrists and hips so common in postmenopausal women and will reduce the mortality presently caused by hip fractures.


Current concepts of rhinoplasty.

Current concepts of rhinoplasty. Like all surgical operation, aesthetic rhinoplasty is a technique that is constantly undergoing change. These changes are brought about by accumulated experience, improved instrumentation, and not the least by the pressure of changing cultural and social requirements. The fine subtleties of rhinoplasty are difficult to learn by the student and difficult to master by the mature surgeon. Technique improves with the accumulation of surgical experience and constant observation and study of one's results. The learning process should never stop. The most significant changes in rhinoplasty technique since the operation was designed are emphasis on attention to detail and less, rather than more, extensive surgery. The removal of less bone, cartilage, and soft tissue is emphasized as well as meticulous attention to details of repositioning and alignment of all structures.


Revision rhinoplasty.

Revision rhinoplasty. Revisionary rhinoplasty is discussed in terms of conceptual organization. Its scope is greater than primary rhinoplasty in that it contains elements of augmentation and reconstruction not seen in the usual primary rhinoplasty. A better general understanding of the relationship of the structural elements to the appearance and the structural elements to the lining is required. Defects encountered in revisionary rhinoplasty reflect the contribution of the three nasal elements, which are the cover, lining, and support. Success depends on accurate evaluation of the distortion produced by each element and their correction. Correction implies maintenance of correct position.


Failure of technetium bone scanning to detect pseudarthroses in spinal fusion for scoliosis. A prospective study of 11 patients suggests that present techniques of technetium bone scanning do not assist in recognizing the presence of well-established pseudarthrosis in spinal fusions for scoliosis.


The anterior heel for metatarsalgia in the adult foot. A conservative approach to foot problems is especially useful in patients with diabetes, rheumatoid arthritis, diminished circulatory changes, and those who are too debilitated for surgical treatment. If one will start with either the medial heel wedge or the anterior heel correction, some response should be evident within 4 to 6 weeks. If the response after a trial period of approximately two to three months for a heel wedge or another two to three months for the anterior heel doesn't relieve pain, then perhaps some other problem might now become easier to localize, i.e. Morton's toe syndrome, hallux valgus, soft or hard corns, or hammertoe deformities. When the anterior heel is prescribed many foot problems other than metatarsalgia will stop being symptomatic and surgery treatment can be bypassed. A physician must know about the supply of shoes in the community and, if necessary, instigate a better inventory of available shoes. In addition, it is necessary to establish good rapport with the shoe repair man so that he will not intrude in your patient rapport or alter your directions. Patients also need advice about losing weight. Frequently a loss of 15 or 20 pounds will change a patient's complaint from one of extremely discomforting daily weight-bearing to a tolerance of a fair amount of walking and at least a reduction of the complaints to a more endurable and functional level. One can't expect the shoe correction to do everything for everyone. The anterior heel isn't the whole solution to the complicated problem but it helps to have patients begin to see results in more comfort in their shoe wear.


The twisted nose.

The twisted nose. To the patient seeking correction of a twisted nose, it is as important to obtain a good shape as it is to achieve normal breathing. The functional problem must be solved regardless of the extent of the resection required. The plastic surgeon may choose among many techniques, some of which have been elucidated. Our preferred approach has been described in detail. We reiterate that experience has shown that a deviated septum which requires stabilization after surgery will recur to a greater or lesser degree at a later date. A twisted nose, to be permanently corrected, should maintain its corrected position without external support at the end of the operation.


Suprascapular nerve entrapment. A report of two cases treated with suprascapular notch resection. Suprascapular nerve entrapment occurs in the shoulder after a severe forward flexing injury (hyperprotraction) because of tethering of the nerve in the scapular notch. Unrelenting pain in this shoulder region is relieved by section of the transverse scapular ligament in most cases. Two patients, reported here, had temporary relief from simple transverse scapular ligament release and later lasting relief of pain from scapular notch resection. These observations suggest that primary wide notch resection may be the preferable procedure.


Practical applications of skin biomechanics. The biomechanical properties of skin have an important influence on plastic surgical decisions. They aid the surgeon in planning elective incisions, excisions, or scar revisions. They provide insight into the most appropriate method of coverage of skin defects as well as the design of an artificial skin substitute. Skin biomechanics can, in part, be characterized in vivo by either a skin extensiometer or by studying the deformation of skin defects. These methods indicate the magnitude and directional orientation of skin tensions which are dependent partly on the mechanical characteristics of the dermal fibers and partly on the pattern in which they are woven. The tensions to which the skin are subjected can be classified as either static or dynamic in origin. Static skin tensions are the natural tensions existing in skin. The magnitude of static tensions varies between individuals, at different sites in the same person, and in different directions in many sites. The dynamic tensions are caused by a combination of forces which are associated with joint movement, mimetic and other voluntary muscle activity, and gravity. Knowledge of these tensions allows the surgeon to align the operative site in the direction of maximal tension and to approximate the wound with the least amount of tension. As a consequence of this, the scar healing between the cut edges of the wound should be narrow and inconspicuous.


Development of cleavage line patterns in the human fetus. The dermal fibroelastic connective tissue of the fetus is arranged in such a manner as to represent cleavage lines (lines of tension). Cleavage lines were observed as early as the 28 mm stage but in most cases they do not appear until the 50 mm stage. The various regions of the body present individual cleavage line patterns which undergo alterations as the fetus grows. The pattern alterations are influenced by (1) rate of proportional growth and (2) principal direction of growth of certain structures in the various regions. The cleavage line pattern of the newborn differs from that of the adult in certain regions.