Differentiating intracerebral neuroblastoma: report of a case and review of the literature.

A case of the rare primary intracerebral neuroblastoma is reported. The patient, a 13-year-old girl, was subjected to craniotomy because of a right frontal tumor. The removed tumor tissue was composed of immature, highly cellular areas with numerous mitotic figures, adjacent to fields with Homer-Wright rosettes, and areas consisting of neuroblasts and mature neurons. There was no evidence of a primary tumor elsewhere in the body and 25 months after the operation and postoperative irradiation, the patient is without any signs of recurrence. Determination of catecholamines and their metabolites in the urine and cerebrospinal fluid gave normal results. Dense-corded vesicles could not be detected in the tumor cells by electron microscopy. A review of previously reported cases of primary intracerebral neuroblastomas is included. The cell of origin and the possibility of transition of a differentiating intracerebral neuroblastoma to ganglioneuroma are discussed.


Adenosine deaminase activity in patients with renal adenocarcinoma.

Erythrocyte and lymphocyte adenosine deaminase (ADA) levels were studied in 31 patients with renal cell carcinoma (RCC). Decreased lymphocyte ADA levels occurred in patients with RCC. Erythrocyte ADA levels were reduced only in blood type B and O patients. Nephrectomy resulted in a rise in lymphocyte and erythrocyte ADA levels. Progression of clinical disease was associated with a fall in lymphocyte ADA values in all patients and with a rise in erythrocyte levels only in blood type A patients. Our results suggest that changes in erythrocyte and lymphocyte ADA levels in RCC patients are acquired and may offer insight into host-tumor interactions.


The quantitative relationship of carcinoembryonic antigen to tumor cells of different organs.

Carcinoembryonic antigen (CEA) was measured in extracts of cell suspensions from tumors of the colon, stomach, breast, lung and kidney. Cell suspensions were also obtained from normal tissue surrounding the primary tumors. The number of neoplastic cells in each cell suspension was determined by cytologic criteria. Highest CEA concentrations were found in cells obtained from carcinoma of the bowel. There also appeared to be a greater difference between the concentrations of CEA in colonic tumors and either surrounding normal tissue or serum, than in tumors of other organs. There was no apparent correlation of the concentration of CEA in tumor cells and the presence or absence of metastases. The presence of CEA in normal cells from tissue adjacent to tumors was suggestive of increased synthesis of CEA by these cells in some cases.


Pseudoleukemia cutis: report of a case in association with molluscum contagiosum.

Histologic sections from a solitary cystic cutaneous lesion that showed atypical mononuclear cells in the dermis and within blood vessels were diagnosed by several general pathologists and dermatopathologists as leukemia cutis. The patient, who had no other cutaneous lesions, was consequently submitted to an extensive investigation for leukemia, which proved negative. Additional and deeper sections from the original block revealed that the cellular infiltrate so suspicious of leukemia cutis was secondary to rupture of a lesion of molluscum contagiosum. The correct histopathologic diagnosis, therefore, was pseudoleukemia cutis. The lessons of the case are that 1) further study of the specimen, solitary as it was and asymptomatic as the patient was, would have obviated worry and the expense and inconvenience of an extensive systemic investigation, and that 2) the diagnosis of leukemia cutis should never be made solely on the basis of histologic sections of skin, but rather after examination of blood and bone marrow.


The bone marrow biopsy, osteoscan, and peripheral blood in non-hematopoietic cancer.

Bone marrow biopsies were obtained from the iliac crests of 101 male patients with unresectable cancers of non-hematopoietic origin. Tumor cells were identified in 31% of the biopsies. Bone scintophotography studies were performed on 67 patients with positive or suspicious scans reported in 67%. Abnormal peripheral blood smears were encountered in 51% of 100 patients. Histologically established marrow metastases associated with a positive osteoscan and an abnormal peripheral smear occurred in 8% of the patients. The association of either one of the latter two with a positive histologic specimen was observed in about one-half of the patients. The significance of these associations is discussed.


Cancer of the esophagus in southern Iran.

The minimal yearly incidence of cancer of the esophagus in Fars Province in Southern Iran, based on hospital records in the capital city, is five to 10 times as high as the yearly incidence in Connecticut for the age group 15-44. Since these represent only a portion of cases among residents of the Province, the true rate is probably considerably higher. An unusual proportion of these cases (32% overall) presented with primary tumor in the upper third of the esophagus. The location of the tumor was related strongly to age, but not to sex after taking age into account. The proportion of esophageal cancer cases who were female was significantly greater than that of other GI tract cancers combined. It appears that the risk in males and females in this area is at least nearly equal, and risk could conceivably be higher in females (similar to the appearance of the disease in Northern Iran).


Hyperparathyroidism in persons treated with X-rays for tuberculous cervical adenitis.

Follow-up examinations of one hundred persons treated with x-rays for tuberculous adenitis between 1930 and 1946 have been carried out to determine if there is an increased incidence of hyperparathyroidism (HPT) after radiation exposure. Neck explorations were done in patients with hypercalcemia and signs and symptoms compatible with HPT. Individuals with thyroid masses were also operated upon when examination of fine needle specimens gave suspicions of malignancy. Eleven subjects were found to have developed parathyroid adenoma or hyperplasia. Four other individuals have hypercalcaemia but are asymptomatic. The mean absorbed dose in the parthyroid glands varied between 75 and 2,200 rads. Six individuals received more than 1,200 rads; four of them later developed HPT, while no HPT occurred below a dose of 300 rads. The high incidence of HPT among patients who had been heavily exposed to radiation suggests a cause and effect relationship between radiation treatment and development of HPT.


Biliary carcinoma associated with biliary cysts.

The 24th reported case of biliary carcinoma associated with biliary cyst is reported and the previously reported cases are reviewed. Analysis of the 24 cases in which carcinoma developed shows that stones were present in the cyst in 25% of cases. The most common cell type was adenocarcinoma (70%) and the most common site for tumor was in the posterior cyst wall (57%). One half of the patients reported developed carcinoma a mean of 4 years following an internal drainage procedure for biliary cystic disease. Of the reported cases of biliary cysts associated with cystlithiasis, one-third had associated biliary carcinoma. It is suggested that excisional therapy be performed when possible in all patients with biliary cystic disease, and in particular, in those with associated cystlithiasis.


Lymphangiomyomatosis with chylous ascites: treatment with dietary fat restriction and medium chain triglycerides.

Lymphangiomyomyoma is a tumor of lymphatic channels and lymph nodes, clinically manifested by recurrent chylous pleural effusions and ascites. The disease is usually progressive and unresponsive to surgery, chemotherapy, or irradiation. A case of lymphangiomyomatosis in a 24-year-old woman who presented with chylous ascites is described. The patient was treated with dietary fat restriction supplemented with medium chain triglycerides and has remained free of recurrent effusions for 2 years.


Endometrial stromal sarcomas.

This study is a clinicopathologic evaluation of seven patients with endometrial stromal sarcoma of pure homologous type. Histologic distinction from endometrial stromatosis was based on the presence of high mitotic activity and nuclear anaplasia. One sarcoma appeared to have originated from invasive stromatosis. All patients died within 27 months although the tumor was judged to have been confined to the uterus at the time of hysterectomy in five patients. The aggressive clinical behavior and high mortality rate documented in this and other studies indicate that stromal sarcoma should be distinguished from the better differentiated and more indolent endometrial stromal tumor known as stromatosis or endolymphatic stromal myosis. It is suggested that surgery be supplemented by irradiation and/or chemotherapy as primary treatment for stromal sarcoma in view of its dismal prognosis.


Ependymoma: a follow-up study of 101 cases.

One hundred and one patients with histologically confirmed ependymomas were studied over a 22-year period. Choroid plexus papilloma and sub-ependymoma were not included. About half of the tumors were intracranial, with the majority of these infratentorial. The intraspinal tumors were equally divided between intramedullary and the "cauda" group. The majority of the intracranial tumors occurred in children, while almost all the intraspinal tumors were in adults. The histologic classification consisted of "typical ependymoma" (cellular, papillary and myxopapillary patterns) and "anaplastic ependymoma". The intracranial and intramedullary tumors showed a predominantly cellular pattern, while the myxopapillary type was found only in the "cauda" group. The histology seems to be of limited value in assessing the prognosis in an individual patient with ependymoma. The postoperative prognosis was poor in the intracranial tumors, although radiotherapy increased the survival time without affecting the eventual fatal outcome. The prognosis in the intraspinal group was much better, with three-fourths of the patients living for at least 10 years. No patient with an anaplastic tumor survived for more than 6 years.


Carcinoma of the breast in women under 30 years of age: a clinical and histopathological study of all cases reported as carcinoma to the Swedish Cancer Registry, 1958-1968.

During 1958, 91 cases of breast carcinoma in women below 30 years of age were reported to the Swedish Cancer Registry. A histologic reexamination showed that 16 of these women had only benign proliferative conditions. Among the remaining 75 patients, two had noninvasive carcinoma, eight advanced carcinoma, and 65 operable invasive carcinoma. The carcinomas were characterized by little fibrous stroma and little or no elastosis. The survival rate of the patients with invasive breast carcinomas was similar to that in a total geographic material of all ages with breast carcinoma. The carcinoma was treated during pregnancy or within 1 year after delivery in 15 patients, with the same survival rate as for those whose carcinoma was not complicated by pregnancy.


A new topographic approach to the spread of breast cancer: the grid method.

In a series of 181 patients with breast cancer treated by mastectomy with axillary lymph node dissection, the authors developed and used an improved topographic technique, which they call the "Grid Method", which maps out the extent of a given cance of the breast. Three types of tumor spread are defined: Type 1, limited (L)(31%); Type 2, multifocal (M)(13.2%); and Type 3, extensive (E)(55.8%). This topographic information allows a more accurate assessment of prognosis when used in conjunction with the morphologic findings. Moreover, the topographic "Grid" study is simple to perform and easy to assess. Of prime importance is the relationship between the topographic and mammographic findings.


A topographic approach to breast cancer: the relation of topographic and mammographic findings.

One hundred and twenty-eight patients with breast cancer who, on the basis of mammographic study, were treated by mastectomy were analyzed. The purpose of this endeavor was to correlate the roentgenologic features with the pathologic findings. The correlation between them was very good: the type of tumor opacity, the microcalcifications, and the various other radiologic patterns were compatible with both localized and extensive breast cancer. Mammography appears to be valuable in the selection of patients suitable for conservative treatment.


The combined effect of breast cancer risk factors.

An extension of the logistic model (Cox, 1970) is applied to the prospective study of breast cancer in Guernsey (Bulbrook and Hayward, 1967). Four important risk factors, age at menarche, family history, age at first birth and etiocholanolone excretion, are identified and shown to have additive effects on a logistic scale for the probability of developing breast cancer. The feasibility of screening a high risk group of women is considered and shown unlikely to be of practical value.


The early detection of colorectal cancer.

A review of the American Cancer Society's statistics for colorectal cancer indicates that there has been little improvements in the survival rate for this disease in the past 25 years. Although there have been advances in surgical techniques, radiation therapy, and chemotherapy, the key to improved survival rates is earlier diagnosis. A significant percentage of patients continues to present with regional or distal metastasis at the time of their initial diagnosis. Both proctosigmoidoscopy and guaiac impregnated filter slide paper methods have been productive in diagnosing this disease at an earlier stage. The "Hemoccult" test, however, is inexpensive, can be used on a routine basis, is easier for patients to perform themselves, and is aesthetically pleasing. It represents a significant cost savings compared to proctosigmoidoscopy and should be advocated for routine use in all patients over 40 years of age. Although there continues to be a role for local and regional mass screening programs, significant improvements in colorectal survival rates cannot be expected until routine application of this type of screening is practiced.


Skin-tumor-initiating ability of benzo(a)pyrene-7,8-diol-9,10-epoxide (anti) when applied topically in tetrahydrofuran.

The skin-tumor-initiating abilities of various metabolites of benzo(a)pyrene (BP) were determined in mice by using a two-stage system of tumorigenesis. We previously reported that BP-7,8-dihydrodiol (+/- trans) was approximately as potent as BP, suggesting that it may be a proximate carcinogen, but the alleged ultimate carcinogen of BP [BP-7,8-dihydrodiol-9,10-epoxide (anti)] was a weak tumor initiator (Cancer Lett.2: 115, 1976). Because of its high reactivity, the tumor-initiating ability of the BP-7,8-dihydrodiol-9,10-epoxide (anti) was determined by using acetone, benzene, and tetrahydrofuran (THF) as the solvent vehicles. The 'diol-epoxide' of BP was found to be an effective tumor initiator when applied topically in THF. The effectiveness of the various vehicles for the 'diol-epoxide' was as follows: THF greater than benzene greater than acetone; however, acetone was the best solvent for BP tumor initiation. The BP-9,10-dihydrodiol and BP-3-hydroxy were found to be weak tumor initiators. BP-3-hydroxy was also tested for tumor-promoting ability and was found to be inactive in this capacity.


Malignant transformation and mutagenesis in mammalian cells induced by vicinal diol-epoxides derived from benzo(a)pyrene.

Benzo[a] pyrene and the syn- and anti-isomers of the 7,8-diol 9,10-oxide and of the 9,10-diol 7,8-oxide derived from this hydrocarbon have been tested for their abilities to induce malignant transformation in M2 mouse fibroblasts and mutagenesis in V79 Chinese hamster cells. The anti-isomer of the 7,8-diol 9,10-oxide induced more mutations and transformation than did the other three vicinal diol-epoxides. The two 9,10-diol 7,8-oxides were moderately mutagenic but did not induce any transformation. In contrast, benzo[a]-pyrene induced transformation in M2 fibroblasts but was not mutagenic in the V79 cells.


Non-epithelial uterine tumours induced in CBA mice by 1,2-dimethylhydrazine.

In CBA mice treated with weekly subcutaneous injections of 1,2-dimethylhydrazine an unusually high incidence of uterine sarcomas was observed in two successive experiments. The tumours are easily transplantable. The description of their histological structure is presented and their histogenesis is discussed.


Acute lethality of D- and L-ethionine in Swiss mice.

The acute, 7-day LD50 in Swiss mice of D-ethionine administered intraperitoneally, was determined as 185 mg/kg with 95% confidence limits of 163 and 210 mg/kg. L-ethionine, the stereoisomer, which is more potent in inhibiting liver RNA synthesis, was not lethal at 2500 mg/kg. This acute toxicity of D-ethionine was suppressed by L-methionine and adenine. Blood accumulation in the thoracic cavity and pericardium was the probable cause of death.


Ultraviolet-induced DNA-repair synthesis in lymphocytes from patients with chronic lymphatic leukemia.

Ultraviolet-induced DNA-repair synthesis was studied in peripheral lymphocytes from 10 patients with chronic lymphatic leukemia (CLL). An ultraviolet-dose curve was established for each patient and compared with curves for healthy individuals. The same level of DNA-repair synthesis was recorded in CLL and normal lymphocytes. This contrasts with a report from another laboratory. Evidence is presented of the necessity of inhibiting replicative DNA-synthesis with hydroxyurea in order to adequately estimate the DNA-repair synthesis.


Distribution of small bowel tumors.

We have studied the anatomic distribution of 784 small bowel carcinomas and have analyzed available epidemiologic data to determine the relation of these rare cancers to other gastrointestinal tumors and to diet. The anatomic distribution of small bowel cancer in unusual: most tumors occur in the duodenum or proximal jejunum, relatively few tumors occur in the central bowel, and only in the lower ileum do the expected number of tumors appear. Review of available epidemiologic data shows a correlation between small bowel cancer and lower, but not upper gastrointestinal cancer. The incidence of small bowel cancer is positively correlated with protein and fat consumption.


Accelerating action of tea on mouse skin carcinogenesis.

The effect of different tannin-containing aqueous solutions on the carcinogenic action of benzo(a)pyrene (BP) on the mouse skin was investigated. It was found that the appearance of tumours was accelerated in mice treated with either tea or an oak extract after a single application of BP. The time of appearance of tumours in mice treated with gallic acid or tannic acid after treatment with BP was similar to that in the group treated with BP alone. The overall incidence of tumours was similar in all five groups.


Whatever happened to NSC--? An analysis of clinical results of discontinued anticancer agents.

Twenty-six investigational anticancer drugs formerly supplied by the National Cancer Institute are no longer available due to the lack of requests for their use in clinical trials. This report examines the data on these drugs to determine how well they were clinically evaluated. Generally, the studies are incomplete, and 34% of the compounds have not been studied beyond phase I trials. Clinical pharmacology data were not obtained for most of the drugs. In addition, the information available for drugs that did undergo phase II trials is grossly inadequate and does not permit a confident decision to withdraw them from investigational use. Instances of "hints" of activity and interesting drug properties are cited to stimulate further study. Finally, a list of compounds scheduled for future termination is given within the framework of this analysis to provoke thought toward obtaining more phase II data before these drugs fall into disuse.


Clinical response and toxicity with cis-dichlorodiammineplatinum(II) in children.

cis-Dichlorodiammineplatinum(II) (DDP) was studied in 16 children with far-advanced malignancies. Three dosage schedules were tried: regimen A, 20 mg/m2/day x 5 days for 3-4 weeks (11 patients); regimen B, 50 mg/m2 once a week (four patients); and regimen C, 60 mg/m2/day x 2 days every 3-4 weeks (one patient). Four of 16 patients (25%) showed partial response, including one with osteogenic sarcoma, one with neuroblastoma, one with seminoma, and one with medullary carcinoma of the thyroid. Two patients showed clinical improvement. The major toxic manifestations included nausea and vomiting (16 of 16), renal toxicity (three of 16), transient pancytopenia (six of 12), and hearing loss (two of 16). It is apparent that DDP has activity in pediatric tumors; however, a more precise response rate must be delineated in a larger series of patients.


Uptake of labeled dianhydrogalactitol into human gliomas and nervous tissue.

The distribution of iv administered 3H-dianhydrogalactitol (DAG) in the plasma, cerebrospinal fluid (CSF), brain, and tumor tissue was studied in 11 patients. DAG entered the CSF and was slowly eliminated, with a half-life of 20 hours. Unchanged DAG accounted for about 6%-30% of the total radioactivity in the CSF. All the tumors accumulated the drug to a higher extent than the intact white matter, except the one meningioma studied. The highest uptake was observed in the relatively benign astrocytic tumors.


Effects of treatment with cyclophosphamide on hormone-dependent and hormone-independent tumor cells in transplanted GR mouse mammary tumors.

Previous studies have indicated that hormone-responsive mammary tumors of GR mice are mixed populations of hormone-dependent and autonomous cells. We have now investigated whether these two cell types differ in susceptibility to cytostatic treatment. Experiments in which cyclophosphamide was injected in tumor-bearing mice did not reveal significant differences in percentage of inhibition between hormone-dependent, hormone-responsive, and hormone-independent tumors. Furthermore, the estrogen and progesterone receptor contents of the residual tumor masses after cyclophosphamide treatment were about the same as those of untreated tumors. When the cytostatically treated tumors were transplanted, the degree of hormone responsiveness of the transplants did not differ from that of transplants derived from untreated tumors, nor did their hormone receptor contents. These results indicate that the hormone-dependent and autonomous cells of GR mouse mammary tumors are inhibited to similar extents by cyclophosphamide. The possible significance of these results for combined endocrine therapy and chemotherapy is discussed.


Changes in physical capacity, heart size and function in patients after myocardial infarction, who underwent a 4-to 6-week physical training program.

From findings in 1,200 coronary patients after myocardial infarction, it was shown that a stationary course of treatment in a rehabilitation center, where a gradually intensifying, individually adapted physical training program is administered, can lead to statistically highly significant improvements in the functional efficiency of the heart, as well as in physical capacity, in a short period of 4-6 weeks. It should be stressed that these positive effects can be demonstrated in the early stages of rehabilitation from infarction, that is, directly following discharge from the hospital, in the same way as in a late course of treatment of the repeated therapy; it is also important to point out that similar favorable results were obtained for older groups as for younger ones under all parameters, within the same time. A 6-week training period is more effective than a 4-week one.


Rehabilitation of the patient after myocardial infarction: the pivotal role of the primary care physician.

Invalidism is often unnecessarily prolonged after myocardial infarction because of misinformation about the natural history of the illness and because of psychosocial difficulties. To correct these problems, the rehabilitative approach should begin at the onset of illness and remain as a continuing feature in the long-term care of the patient; rehabilitation must thus be the responsibility of the patient's primary physician, who may be a general physician or a practicing cardiologist. Cardiovascular rehabilitation services should be designed to restore the patient, as rapidly as possible, to a productive, active and satisfying life. For patients with significant residual cardiac damage or particular psychological social or vocational problems, conventional medical management is often insufficient to ensure return to a full, active and productive life. The clinician should appreciate this problem early in the course of the illness and institute referral for specialized consultant assessment and management. However, all aspects of the long-term care and rehabilitation of the patient after myocardial infarction must remain under the aegis of the primary physician, ultimately using the personnel, facilities and services of the local community.


Infarct sizing with technetium-99m stannous pyrophosphate scintigraphy in dogs and man; relationship between scintigraphic and praecordial mapping estimates of infarct size in patients.

The present study was performed in order to evaluate the ability of technetium-99m stannous pyrophosphate (99mTc-PYP) myocardial scintigrams to size infarcts in experimental animals and man. In 10 dogs with proximal left anterior descending coronary artery occlusion and acute anterior myocardial infarcts, there was a significant correlation between scintigraphic infarct size and histological infarct weight (P less than 0.01). In 25 patients with acute anterior or anterolateral myocardial infarcts, there was a significant correlation between relatively large infarct size determined scintigraphically and the acute development of left ventricular failure. There was some overlap, however, in 99mTc-PYP scintigraphic infarct size between patients who did and did not develop left ventricular failure with infarction. Presumably this is explained by some patients having had earlier myocardial damage and thus developing left ventricular failure with relatively small new infarcts. There was also a statistically significant, but weak, correlation in patients between scintigraphic infarct size and precordial ST segment mapping including peak ST segment elevation (P less than 0.05) and the number of praecordial sites with ST segment elevation equal to or greater than 2 mm (P less than 0.01). The data suggest that 99mTc-PYP scintigrams and praecordial mapping measure some similar but some dissimilar aspects of infarct size in patients, and that 99mTc-PYP scintigraphy does size acute anterior and anterolateral infarcts in experimental animals and patients.


Effect of regional myocardial ischaemia on left ventricular isovolumic contraction.

In anaesthetised, open-chest dogs, global isovolumic contractility indices were reduced by acute regional ischaemia involving 12 to 14% or more of left ventricular mass. These effects were not altered by controlled reduction of blood flow to the non-ischaemic region, but were exaggerated by isoprenaline during normal perfusion of the non-ischaemic region. The results were consistent with a contraction model characterised by simple reduction of the effective length of the contractile elements.


Cardiovascular and antiarrhythmic effects of aprindine (AC1802) during partial occlusion of a coronary artery in the pig.

The effect of intravenously administered aprindine (AC1802) as a prophylactic agent against ventricular arrhythmias was studied in pigs. During the first 30 min of ischaemia 5 of the 22 untreated animals died because of ventricular fibrillation against 1 of the 23 animals pretreated with aprindine (P=0.09). Ventricular tachycardias were observed in 10 untreated animals and in none of the aprindine group (P=0.0002). The incidence of other arrhythmias was significantly less in the aprindine group compared with the untreated group (P less than 0.02).


Calcium kinetics in the aorta of spontaneously hypertensive rats.

In the search for abnormalities in hypertensive vessels Ca kinetics was studied in the aorta of spontaneously hypertensive rats (SHR). The uptake of 45Ca was significantly less in hypertensive animals but the uptake of 'Lanthanum resistant' 45Ca was not. The efflux of 45Ca from aorta of SHR was different (P less than 0.01) from that of control rats because of faster initial efflux. The half-life of the first 2 components of efflux curves was significantly shorter in SHR. Results are interpreted as consequences of decreased Ca binding by smooth muscle membrane in hypertensive vessels.


Right and left ventricular compliance in the hereditary cardiomyopathy of the Syrian hamster.

Ventricular pressure-volume curves were examined in 10 pre-oedematous cardiomyopathic Syrian hamsters, aged 120 d, and 10 oedematous cardiomyopathic hamsters, aged 210 d, and compared with 10 and 8 age-matched controls, respectively. Previous studies had shown filling pressures and cardiac output to be normal in the pre-oedematous stage. In contrast, the oedematous stage was characterised by elevated filling pressures and increased cardiac output, raising the question whether this stage represents true myocardial failure or circulatory congestion associated with decreased ventricular compliance. Compliance, defined as dV/dP, was derived simulataneously for both ventricles from post-mortem pressure-volume curves from 0 to 2.66 kPa (0 to 20 mmHg). Left ventricular tissue elastic modulus, E, defined as dsigma/depsilon or the incremental stress (sigma) for an increment of strain (epsilon), was derived from the compliance curves and certain linear dimensions of the heart. At isobaric intervals, compliance did not differ between control and myopathic ventricles. At pressures corresponding to previously measured, naturally prevailing end-diastolic pressures, the myopathic ventricles were significantly dilated in both stages; end-diastolic compliance was normal in pre-oedematous hamsters, but was significantly decreased in the oedematous animals. Left ventricular elastic modulus was elevated at all values of sigma in both groups of myopathic animals.


The cornified envelope of terminally differentiated human epidermal keratinocytes consists of cross-linked protein.

A small proportion of the protein of stratum corneum of human epidermal callus is insoluble even when boiled in solutions containing sodium dodecylsulfate and a reducing agent. This protein is present in the cornified envelope, a structure located beneath the plasma membrane. When cornified envelopes were dissolved by exhaustive proteolytic digestion and the products analyzed by chromatography, approximately 18% of the total lysine residues were found as the cross-linking dipeptide epsilon-(gamma-glutamyl) lysine. Labeled cornified envelope protein was synthesized by human epidermal keratinocytes allowed to differentiate terminally in culture. The extent of cross-linking, determined from the proportion of radioactive lysine in epsilon-(gamma-glutamyl) lysine after exhaustive proteolysis, was similar to that in stratum corneum. The properties of the cornified envelopes (insolubility in detergent and reducing agents, and solubility following proteolytic digestion) are readily explained by a structure consisting of a cross-linked protein lattice.


Zea mays chloroplast ribosomal RNA genes are part of a 22,000 base pair inverted repeat.

Zea mays chloroplast rDNA exists in two identical units. Each unit contains one sequence for the 16, 23 and 5S rRNAs in the order given. The 16 and 23S sequences in each unit are separated by a 2100 base pair (bp) spacer. The DNA sequence for 5S RNA is closely linked to that for the 23S RNA. Within the above unit, the three RNAs are transcribed from a single DNA strand. The two rDNA units on the circular chloroplast DNA molecule are separated from each other by 18,500 bp in one direction and by 106,100 bp in the other direction. The two rDNA units have an inverted orientation with respect to each other. Each rDNA unit is part of a 22,000 bp sequence which is repeated with inverted orientation.


Binding of [125I]concanavalin A by interspecific amoeba hybrids.

Two types of interspecific amoeba hybrids, viz. PnIc (Amoeba proteus nucleus in A. indica cytoplasm) and InPc (A. indica nucleus in A. proteus cytoplasm), were tested for their [125I]Concanavalin A (Con A) binding activity at different periods of time. The cell surface binding of the labelled Con A was reduced to approx. 40 and 75% in the PnIc and InPc cells, respectively, 96 h after the cells were made hybrids. A significant increase in the binding of radioactive Con A was observed after the homologous nucleus was back transferred into InPc cells, but not in PnIc cells. No significant difference in the binding of the radioactive Con A was found between the progeny of the hybrid amoebae and the control amoebae. The Con A binding activity of the interspecific amoebae has been discussed in relation to the changes in the cytoplasmic membrane structures which presumably play a role in the formation of cell surface in the amoebae.


Glycosaminoglycan and glycoprotein synthesis by cranial neural crest cells in vitro.

In the developing chick embryo, cranial neural crest cells, that will subsequently give rise to facial mesenchyme tissues, migrate beneath the surface ectoderm in a cell-free and hyaluronate-rich matrix. To determine how the crest cells could contribute to this matrix, we cultured crest cells from stage 9 embryos for 2 days and then labeled them for 18--20 h with various precursors of glycosaminoglycans (GAG) and glycoproteins. [3H]fucose and [3H]glucosamine were incorporated into Pronase-sensitive macromolecules associated with the cell layer, but little labeled glycoprotein was released into the medium. Hyaluronate was the major GAG synthesized and was distributed between the cells and medium. Less chondroitin-sulfate was synthesized. In comparison, older cultures as well as fibroblasts produced different proportions of GAG. Our results confirm the autoradiographic findings of Pratt et al. (1975) and suggest that crest cells may contribute GAG to the matrix during migration.


Studies of protein synthesis during sea urchin oogenesis. II. Synthesis of tubulin.

Sea urchin oocytes were incubated in vitro with [14C]lysine. Tubulin synthesis was demonstrated by gel filtration and/or electrophoresis of the colchicine-tubulin complex precipitated by vinblastine. The incorporation of [14C]lysine into tubulin represented about 12% of the incorporation into the total proteins at vitellogenesis.


DNA replication in human diploid cells of different origin.

DNA-fiber autoradiography has been used to study the replication of chromosomal DNA from cultured human cells of different origin. In normal embryonic fibroblasts, brain cells, heart cells, hepatocytes, and post-natal fibroblasts and lymphocytes the rate of DNA replication in a single fork varies from 0.2 to 1.0 micrometer/min with a mean of about 0.6 micrometer/min; and the size of replication units varies from 80 to 400 micrometer with a mean of about 200 micrometer. The data obtained indicate that in cultured somatic cells of various tissues of the same organism, the rate of DNA replication and the number of functioning replication units are practically the same.


Some effects of chlorambucil on nuclear protein phosphorylation in the Yoshida ascites sarcoma.

The ability of nuclei, isolated from Yoshida ascites sarcoma cells, to phosphorylate nuclear proteins in the presence of [gamma-32P]ATP has been investigated. Comparisons were made between a strain sensitive to the effects of the alkylating agent, chlorambucil, with a corresponding resistant strain both before and after drug-treatment of tumour-bearing animals. There was no gross quantitative differences between the drug-sensitive and-resistant untreated cells but treatment resulted in increased levels in the sensitive strain. Qualitative differences were seen before treatment in the phosphorylation pattern of the tris-saline-soluble nuclear sap fraction. The high molecular weight species in the fraction from sensitive cells showed phosphorylations which were absent, or present at very low levels, in the corresponding fraction from drug-resistant cells. Changes were observed in the tris-saline-soluble and non-histone protein fractions from drug-sensitive cells following treatment of tumour-bearing animals. Only minor alterations in patterns of phosphorylation were seen in fractions from drug-resistant cells.


Biological effects of 2,4-pentadiene-1-ol.

The biological effects of 2,4-pentadiene-1-ol (CH2 = CH--CH = CH--CH2--OH) the starting monomer in the preparation of a biocompatible polymer were studied. The experiments in rats revealed both a high local (oedema, necrosis) and systemic toxicity. Within 5-10 min after dermal application of 0.2 ml the compound evoked intensive hyperaemia of acral parts (ears, snout, paws) and induced severe breathing difficulties which led to death in half of the animals after a single administration. The most evident macroscopic and microscopic pathologic changes were found in the place of application and in the lungs (haemorrhage in alveolae). Both clinical and histological findings indicated that the toxicity is due to an increased permeability of capillaries.


Subcellular distribution, a factor in risk evaluation of pentachlorophenol.

Pentachlorophenol (PCP) is a potent uncoupler of mitochondrial phosphorylation in vitro and also interferes with microsomal detoxication functions in vitro. This favours flavin mediated oxygenation compared with flavin cytochrome P-450 dependent reactions. Gas chromatographic analysis of subcellular fractions, obtained by zonal centrifugation showed markedly lower PCP concentration in mitochondria and a high accumulation in microsomes compared with cytosol. This increases the likelihood that PCP in vivo causes a malfunction in microsomal detoxication.


Disturbance of microsomal detoxication mechanisms in liver by chlorophenol pesticides.

The pesticide pentachlorophenol known as an uncoupler of mitochondrial oxidative phosphorylation was shown to disturb liver microsomal detoxication functions by a selective inhibition of the terminal oxygenation enzyme P-450. At lower concentrations the flavin moiety of this enzyme chain is not inhibited but rather is stimulated, whereby a qualitative shift in detoxication of aromatic amines from C-oxygenation to N-oxygenation is obtained. The effects were due to the pentachlorophenol itself and not to a metabolite. Similar effects of varying strength were also obtained with other chlorophenol pesticides; 2,4,di-, 2,4,6,-tri and 2,3,4,6-tetrachlorophenol, di- and hexachlorophen, tri- and nonachloro-2-hydroxydiphenyl ethers. The relevance of these findings to the possible synergistic influence of chlorophenols on the carcinogenic effects of polyaromatic amines and hydrocarbons is discussed.


The metabolism and antitumour activity of the enantiomers of cis- and trans-4-methylcyclophosphamide.

4-Methylcyclophosphamide, an analogue of the antitumour agent cyclophosphamide, exists in cis and trans forms, each of which comprises a pair of optical isomers. The extents of metabolism by rat liver microsomes during 20 min were compared for the four steroisomers incubated separately, and for the racemic cis and trans-derivatives in admixture, using mass spectrometry and gas chromatography respectively. In comparative antitumour tests against the ADJ/PC6 plasma cell tumour in mice, the racemic cis and trans forms of 4- and 6-methylcyclophosphamide had similar therapeutic indices. The four stereoisomers of 4-methylcyclophosphamide exhibited an approx. two-fold range in therapeutic index so that there was no marked effect on either metabolism or antitumour activity occasioned by change of configuration either at C-4 or at phosphorus.


Effect of aflatoxin B1 on hepatic polyribosomes and protein synthesis in the rat.

This study was concerned with the effect of aflatoxin B1 on protein synthesis in rat liver. Specifically, the effect of the administration of aflatoxin B1 (6.0 mg/kg body weight) on hepatic polyribosomes (free and membranebound), protein synthesis in vitro, and initiation factors activity of rats within 3-12 h was investigated. The results revealed that aflatoxin B1 rapidly led to disaggregation of free and membrane-bound polyribosomes, to inhibition of in vitro protein synthesis by both populations of polyribosomes, to little or no effect on the activities of initiation factors, and to defective ribosomes, particularly the 60S ribosomal subnits, of both types of polyribosomes. Comparative studies on the effects of aflatoxin B1 and actinomycin D revealed progressive disaggregation by both hepatic free and membrane-bound polyribosomes after aflatoxin B1 but not only of the free polyribosomes after actinomycin D.


Isolation of a non-thionein copper-binding protein from liver of copper-injected rats.

1. The characterization of a low molecular weight, non-thionein, Cu-binding protein isolated from rat liver is reported. The protein was isolated following chronic administration of Cu(NO3)2 using a combination of Sephadex G75 and Sephadex DEAE A-25 chromatography. The protein did not bind to fully equilibrated Sephadex DEAE which formed the basis of the isolation procedure. 2. The final protein pereparation was found to be homogenous by a variety of electrophoretic techniques and was distinguished from metallothionein on the basis of its behaviour on ion exhcange and electrophoretic systems, spectral properties, and amino acid composition and metal content. It contains 6.8% cysteine and was found to bind Cu in a ration of 1.5:1 based on a molecular weight of 11 000. 3. These results confirm the necessity to use techniques other than gel filtration alone to obtain adequate separation of low molecular weight metal-binding protein fractions.


Benzo(a)pyrene metabolism in mouse epidermis. Analysis by high pressure liquid chromatography and DNA binding.

Mouse epidermal homogenates contain an inducible aryl hydrocarbon hydroxylase (AHH) complex that catalyzes the formation of benzo(a)pyrene-7,8-dihydrodiol from benzo(a)pyrene (BP) as assessed by high pressure liquid chromatography (HPLC). 5,6-Benzoflavone (5,6-BF), 7,8-benzoflavone (7,8-BF) and 17-beta-estradiol decreased and butylated hydroxytoluene (BHT) enhanced oxidative metabolism of BP when added in vitro. Epoxide hydrase activity (hydration of benzo(a)pyrene-4,5-epoxide) (BP-4,5-epoxide) was enhanced by 17-beta-estradiol, 5,6-BF, and 7,8-BF. BHT exhibited no significant effect and 1,2-epoxy-3,3,3-trichloropropane (TCPO) inhibited hydrase activity. The capacity of epidermal homogenates to catalyze the covalent binding of BP to DNA indicated that addition of both 5,6-BF and 7,8-BF decreased binding. BHT and TCPO did not significantly affect DNA-binding.


Action of monovalent cations on the biological properties of PR toxin, a mycotoxin from Penicillium roqueforti.

PR toxin impairs liver cell metabolism by inhibiting RNA and protein synthesis. In vitro, the drug inhibits the transcription carried out by isolated rat liver nuclei and the translation promoted by polysomes. The action of monovalent cations on the biological activities of PR toxin has been studied. The increased ionic strength due to the presence of salt in the incubation medium, lowers the inhibitory action of PR toxin on in vitro transcription and translation activities; this action is reversible. Besides the overall effect of the ionic strength, ammonium salts possess a specific ability to suppress irreversibly the biological properties of PR toxin (in vivo toxicity and capacity of inhibiting RNA and protein synthesis). The mechanism of this action is discussed.


The metabolic activation of 7-methylbenz(a)anthracene in mouse skin.

The metabolism of 7-methylbenz(a)anthracene by rat-liver preparations and by mouse skin has been studied using a combination of thin-layer and high pressure liquid chromatography and all five possible trans-dihydrodiols have been detected as metabolites but in different proportions. The roles of these dihydrodiols and of the related vicinal diol-epoxides in the metabolic activation of 7-methylbenz(a)anthracene in mouse skin has been studied using Sephadex LH-20 column chromatography. The results show that the hydrocarbon-nucleic acid products formed in mouse skin in vivo most probably arise from 3,4-dihydro-3,4-dihydroxy-7-methylbenz(a)anthracene 1,2-oxide which, on the basis of this and other evidence, appears to be the reactive intermediate involved in the metabolic activation of 7-methylbenz(a)anthracene in this tissue.


Alkylated purines in the DNA of various rat tissues after administration of 1,2-dimethylhydrazine.

[1,2-3H] Dimethylhydrazine (1,2-DMH) (300 mg/kg; 4 mCi/kg) was injected s.c. to BD-VI rats and the degree of DNA alkylation was measured in various tissues 3 h after the administration. The O6-methylguanine/7-methylguanine ratio was approx. 4 times higher in the colon's DNA (0.0565), the principal target organ of this carcinogen, than in the liver's DNA (0.0136). These findings are briefly discussed in relation to the organ specificity of this carcinogen.


Synthesis of phosphonate and ether analogs of rac-phosphatidyl-L-serine.

The chemical synthesis of four phosphonate-containing phosphatidylserine analogs namely, L-serine (+/-)-[2,3-bis(hexadecyloxy) and 2,3-bis(Palmitoyloxy)-propyl] phosphonates, and L-serine (+/-)-[3,4-bis(hexadecyloxy and 3,4-bis(palmitoyloxy)-butyL]phosphonates is descirbed. (+/-)-2,3-Bis(hexadecyloxy) and 2,3-bis(palmitoyloxy)-prophylphosphonic acids and (+/-)-3,4-bis(hexadecyloxy)butylphosponic acid were prepared by reaction of tris(trimethylsily) phosphite on the corresponding haloalkane. Condensation of the above phosphonic acids or (+/-)-3,4-bis(palmitoyloxy)butylphosphonic acid with N-carboxy-L-serine dibenzyl ester in the presence of trichloroacetonitrile or triisopropylbenzenesulfonyl chloride yielded the protected serine intermediates, which on hydrogenolysis gave the desired L-serine analogs. By a similar route, 1,2-dihexadecyl-rac-glycero-3-phosphoric acid was converted to 1,2-dihexadecyl-rac-glycerophospho-L-serine (L-serine (+/-)-2,3-bis(hexadecyloxy)propyl hydrogen phosphate(ester).


Cholesteryl sulphate and phosphate in the solid state and in aqueous systems.

Cholesteryl sodium sulphate (CS) crystallizes as the dihydrate, the crystal structure of which is known. On heating the dihydrate, solid state phase transitions are observed at 65 degrees C and 95 degrees C and melting occurs at 165 degrees C. The structure of the high temperature phase has not been determined. Cholesteryl dihydrogen phosphate (CP) is not isostructural with any phases of CS. It undergoes a phase transition at 50 degrees C and melts at 190 degrees C. In systems with water CS is unstable whereas it was possible to determine the phase diagram of CP. In most of the composition range a crystalline hydrate is in equilibrium with a gel-phase. The latter has remarkable properties in that lamellar order exists with the 46 A lipid bilayer interleaved with water layers up to 1000 A. The monofilm behaviour of CS and CP at different pH levels is also reported.


Phosphonosphingoglycolipid, a novel sphingolipid from the viscera of Turbo cornutus.

A novel lipid which contained long-chain base, fatty acid, galactose and N-methylamino-ethylphosphonic acid in an equimolar was isolated from the viscera of Turbo cornutus. The method used for the structural elucidation of this lipid were partial acid hydrolysis, alkaline hydrolysis, periodate oxidation and Smith degradation. The structure of break-down products were mainly identified by combined gas chromatography and mass spectrometry. The structure of the novel lipid was determined to be 1-O-[6'-O-(N-methylaminoethyl-phosphonyl) galactopyranosyl] ceramide. Mass spectra of galactose-N-methylaminoethylphospnate and glycerol-N-methylamino-ethylphosphonate are given.


Interactions between fluorescent labeled phosphatidyl serine and cations.

Phosphatidyl serine (PS) was reacted with dimethylaminoanphthalenesulfonyl (DNS) chloride to produce a fluorescent phospholipid derivative (DNS--PS). By comparison of the emission maximum of the fluorochrome in solvents of varying dielectric constants and in aqueous suspension it was concluded that the DNS moiety of sonicated DNS--PS micells was within the glycerol region of the lipid. Addition of CaC12 to aqueous suspensions of DNS--PS micells caused a shift in the emission maximum from 524 nm to 490 nm, a two-fold increase in the relative fluorescence and an increase in the polarization of fluorescence and finally aggregation. MgC12 produced similar but smaller changes in the fluorescence parameters. In the presence of calcium the thermal transition of the fatty acid tail region could be monitored as an increase in the fluorescence intensity of the DNS group. Without calcium an almost linear decrease in relative fluorescence was observed. These data suggested that calcium caused the fluorochrome to be shifted from the glycerol region of the lipid structure into the fatty acid tail region. These observations indicate how the conformation of a biological membrane might be altered by the interaction between acidic phospholipids and calcium.


The interaction of lanthanide and calcium salts with phospholipid bilayer vesicles: the validity of the nuclear magnetic resonance method for determination of vesicle bilayer phospholipid surface ratios.

Contrary to a recent report (B. Sears et al., Biochemistry 15 (1976) 1635), it has been determined that the ratio of the number of phospholipids on the inner and outer surfaces of phospholipid bilayer vesicles can be accurately determined by NMR paramagnetic ion shift reagent studies of vesicles. It is concluded that the metal interacts with all of the phospholipid on the exposed bilayer surface. A ratio of outer phospholipid to inner surface phospholipid of 2.1 +/- 0.1 is obtained regardless of the nucleus studied, position of the nucleus relative to the metal ion binding site, molar ratio of metal to phospholipid over three orders of magnitude, or location of the metal ion of the inside or outside of the vesicle. Additionally, P-31 NMR studies using LaC13 and CaC12 indicate that Ca2+ weakly interacts with egg PC vesicles and that the lanthanides are adequate substitutes for Ca2+ since neither metal is found to perturb measurably the average polar head group conformation.


Dynamics of phospholipid aggregation in ethanol--water solutions.

The kinetics of liposome and vesicle formation of a synthetic lecithin has been studied by light scattering techniques. It is shown, that by evaporation of alcohol from a lipid--alcohol--water mixture, the aggregates formed undergo several changes in shape. A hypothesis is presented, visualising the formation of liposomes or vesicles from monomers, which is consistent with the experimental observations.


[Cytobiopsy as the best present method of intraoperative diagnosis of tumors of the pancreas].

A report has been made over six years experience with a fine needle aspiration biopsy, from authors who have developed a cytological intraoperative quick diagnosis of pancreas tumours. 116 patients have been treated up to this time. This method has proved to be important for the diagnosis of small pancreas tumours, while especially in such cases the indication to pancreas resection is used only in case of malignant tumours. In relation to other normal histological freezer microtome, the cytological method of fine needle biopsy has the advantage of being completely without complication. One can make as many punctures as needed and exactly this is what assues accurate diagnosis. In 90 cases with certain diagnosis, 65 correct conclusions and 12 suspect conclusions of tumour were obtained, and only five cases were erroneously judged negative; in eight cases the poor material did not allow a diagnosis. Therefore, wide application of cytobiopsy can be recommended.


[Total colectomy in the rat : its metabolic and functional consequences].

A number of total colectomies with end-to-end ileo-rectal anastomosis was performed in the inbred Lister rat. The following data were studied: survival rate, body weight curves, variations of appetite, thirst and diuresis, daily weight and electrolytes content of feces, intestinal transit time, histology of terminal ileum, rectum, liver and kidney. After an initial lose of body weight, rats gained their original weight at about two months after colectomy; at this time a syndrome was observed caracterized by anemia, leucocitosis with neutrophilia, ipoproteinemia, remarkable increase of thirst (up to 400%) and of water and sodium content of feces; appetite and diuresis were normal; intestinal transit tice was slightly reduced. Histologically there was a "coloni" transformation of the terminal ileum and vacuolization of the proximal renal tubules.


[Serum gastrin levels in patients with recurrent peptic ulcer].

Serum gastrin behaviour after stimulation with a protein meal was examined in 20 patients of both sexes suffering from x-graphically and endoscopically accertained recurring peptic ulcer, and a marked increase was found in the hormone in the majority of patients when observations were made 15 and 45 minutes after the meal. The genesis of this increase is therefore discussed, with consideration of the importance that extra-gastric sites of production of the hormone may have in such penomenon.


[Metabolic and histological changes induced in the rat by massive resection of the small intestine].

The aim of this paper was o study the long-term changes induced in the rat as a consequence of a 90% resection of small intestine. At 6 months after operation the surviving rats were stable and showed: hypoproteinemia, hyperlipemia, and a decrease of bile flow, total lipids, and cholesterol of bile. What residual intestine concern there was an increase in length and size, and decreased transit time; stomach, duodenum and colon too showed increase size. Histologically there was hypertrophy of villi, regressive changes of pancreas and kidney, as well as deposit of iron in the kidney. The reasons for these changes are discussed.


[Surgical treatment of neoplasms of the vestibule of the mouth].

In this work the Authors show their own operative technique, attained in the surgical treatment of the neoplasms of the vestibule of the mouth. This operative technique also shows the possibility of reducing two surgical treatments to one. In other words it is possible to remove the primitive lesion and carry out, at the same time, the emptiness of those lymphonodulos chains the lymphtic vessels of the region are tributary to. In such a way the time elapsed between the operation and the following nuclear therapy is considerably reduced. Besides, though the exeresis implies an aesthetical injury, not considerable anyway, no functional deficiencies are shown since all the mouth functions, mastication, phonation, language, deglutition etc. are promptly recovered.


Effects of prostaglandin inhibition on intrarenal hemodynamics in acutely saline-loaded rats.

We studied the effect of inhibition of the prostaglandin (PG)-synthesizing enzyme system in female Sprague-Dawley rats following acute expansion of the extracellular fluid volume (ECV). In 57 conscious rats expansion of the ECV with isotonic saline corresponding to an increase in body weight of 10% was induced. Prior to ECV expansion 31 rats received indomethacin (10 mg/kg of body wt) by stomach tube. In six non-ECV-expanded rats indomethacin had no effect on glomerular filtration rate (GFR) and renal plasma flow (RPF). In ECV-expanded rats pretreated with indomethacin, GFR was unaltered but 125I-hippuran clearance decreased, and filtration fraction significantly increased. Intrarenal 86Rb distribution was similar in control and ECV-expanded rats. Indomethacin caused a slight increase in relative cortical 86 RB activity in non-ECV-expanded rats, but had no effect on intrarenal 86Rb distribution in ECV-expanded rats. No difference in intracortical glomerular perfusion was noted between control and ECV-expanded rats. In indomethacin-treated ECV-expanded rats an increase in relative inner cortical perfusion was observed. Absolute perfusion remained unaltered. Thus the decrease in total RPF was entirely due to decreased perfusion of outer cortical nephrons. Renal prostaglandins therefore may play a permissive role for physical factors to promote renal sodium excretion in acute ECV expansion via changes in intrarenal hemodynamics.


Effect of angiotensin-converting enzyme inhibitor (SQ 20881) on the plasma concentration of angiotensin I, angiotensin II, and arginine vasopressin in the dog during hemorrhagic shock.

The effect of an angiotensin-converting enzyme inhibitor on the circulating levels of angiotensin I, angiotensin II, and arginine vasopressin was studied in dogs subjected to hypotensive hemorrhagic shock. In dogs subjected to hemorrhage but not given the inhibitor, angiotensin II rose 20-fold (from 69 to 1,343 pg/ml of plasma), whereas in dogs subjected to hemorrhage but pretreated with the inhibitor, angiotensin II rose only 2-fold (from 92 to 171 pg/ml of plasma). In the pretreated dogs angiotensin I rose 30-fold (from 108 to 3,232 pg/ml of plasma). There was no statistically significant difference between the vasopressin levels found in the untreated dogs and the levels found in dogs given the inhibitor (1,016 and 1,095 pg/ml of plasma). Of the 15 dogs in the untreated group, five died before retransfusion was completed (four of cardiac failure and one of cardiac arrhythmia); none of the 10 dogs in the inhibitor-treated group died. These observations suggest that the very high levels of angiotensin II observed following severe hemorrhage do not contribute significantly to the increased secretion of vasopressin and that the inhibitor protects against death, possibly by suppressing the very high blood levels of angiotensin II observed following this type of experimental hemorrhagic shock.


Elastin and collagen accumulation in rabbit ascending aorta and pulmonary trunk during postnatal growth. Correlation of cellular synthetic response with medial tension.

Absolute and relative quantities of elastin, collagen, and DNA in anatomically defined segments of rabbit ascending aorta (AA) and pulmonary trunk (PT) were compared at intervals from birth to 2 months of age. Identical in size, weight, and composition at birth, the vessels maintained similar lengths and diameters at each age but diverged markedly in weight and scleroprotein content after 1 week. By 2 months, 3 times as much elastin and 1.7 times as much collagen had accumulated in the AA as compared to the PT. By contrast, the increase in total DNA content was the same for both segments. Differences in total fibrous protein accumulation, total elastin accumulation, and elastin content relative to DNA paralleled differences in estimated total medial tangential tension. Proportions of elastin and collagen relative to dry weight increased markedly only between 4 and 2 weeks of age and not thereafter despite continuing rapid growth, steadily increasing medial tension, and increasing total scleroprotein content. Thus, medial cells were capable of adapting their quantitative scleroprotein synthetic response to differences in medial tension throughout growth but established a fixed qualitative response within 2 weeks.


Effect of hyaluronidase and methylprednisolone on myocardial function, glucose metabolism, and coronary flow in the isolated ischemic rat heart.

Ischemia in the isolated perfused rat heart resulted in an increase in coronary vascular resistance. Studies were undertaken to determine the effect of hyaluronidase and methylprednisolone on this increase in resistance as well as on glycolytic rate and mechanical function of ischemic hearts. Neither hyaluronidase nor methylprednisolone affected the rate of glucose utilization in working perfused control or ischemic rat hearts. However, both agents prevented a reduction in coronary flow during a 2-hour ischemic period. Associated with the higher coronary flows were higher tissue concentrations of creatine phosphate and lower concentrations of lactate. These agents also prevented accumulation of tissue water in the ischemic hearts. Such changes would appear to be beneficial to the ischemic heart, although mechanical function of post-ischemic hearts was not enhanced by the presence of either hyaluronidase or methylprednisolone. The results, however, suggest that the reduction in myocardial infarct size noted with hyaluronidase and methylprednisolone may be due to their prevention of further reduction of coronary flow in marginally eschemic tissue.


Quantitative studies of microcirculatory structure and function. III. Microvascular hemodynamics of cat mesentery and rabbit omentum.

We made simultaneous measurements of intravascular pressure and red blood cell velocity for vessels which make up the modular configuration of microvascular networks in mesentery and omentum. An analysis of these variables and the computed volumetric flow rates is presented for arterioles which had a maximum diameter of 56 micrometer through the "true capillaries" (typically 7 micrometer for mesentery and 8 micrometer for omentum) to 56-micrometer venules. The spatial variance of pressure and flow is related to topographical features of each network. Vascularization statistics for each network are presented and demonstrate a unique ratio of potential microvascular exchange area to module planar area, with values of 0.71 +/- 0.22 (SD) for omentum and 0.19 "/- 0.03 (SD) for mesentery. Analysis of the volumetric flow rate for each module demonstrates a linear relationship to the planar area of tissue serviced by each modular network. In situ perfusion rates of 1180 ml/min per 100 g and 105 ml/min per 100 g were determined for omentum and mesentery, respectively. The hemodynamic resistance of the omental and mesenteric circuitry was evaluated, and in the case of the omentum, found to be inversely proportional to the planar area of the module. The arterial to venous distribution of pressure and flow for the mosaic of contiguous modules in omentum and mesentery is described and related to the deployment of parallel and serial microvessels of each network.


Wall shear stress distribution in a model canine artery during steady flow.

The wall shear stress pattern was measured in a rigid plastic cast of a canine artery during steady flow by means of an electrochemical technique. The topographic distribution of shear stress is very nonuniform, with regions of high and low shear in close proximity. The steady shear stresses are highest at the leading edge of flow dividers and at the entrance regions to branch vessels. The shapes of the shear stress curves in the celiac branch are primarily a function of the ratio of branch flow to total aortic flow. However, the shapes of the shear stress curves in the adjacent anterior mesenteric branch remain the same for different anterior mesenteric branch flow ratios, although the shear increases with the branch flow ratio. An unstable pattern of flow separation and reattachment is found at the anterior mesenteric flow divider lip and remains localized to that region. A correlation is suggested between sites of high shear stress, extremes in the range of stress, and unstable stress patterns and sites at which atherosclerosis has been shown to develop.


Some observations on the kinetics of the Jaffé reaction for creatinine.

The Jaffé reaction for creatinine assay appears to follow pseudo-first-order kinetics; first-order rate constants are different for different samples. Rate constants for 10 different serum samples varied from a low value of 0.0040 +/- 0.0003 s-1 to 0.0084 +/- 0.0008 s-1. We describe an approach for determining first-order rate constants from kinetic data and discuss the effects of the above observations on the mathematical formulations required for reliable kinetic determinations of creatinine.


Enzyme-coupled measurement of uric acid in serum with a centrifugal analyzer.

An early-reading blank-corrected end-point determination of uric acid in serum has been developed for use with a centrifugal analyzer. The method is based on a modification of the uricase (urate:oxygen oxidoreductase, EC 1.7.3.3)/catalase (hydrogen peroxide:hydrogen peroxide oxidoreductase EC 1.11.1.6)/aldehyde dehydrogenase (aldehyde:NAD(P)+ oxidoreductase, EC 1.2.1.5)-coupled analytical scheme reported by Haeckel [Z. Klin. Chem. Klin. Biochem. 14, 101 (1976)]. Sensitivity and precision of the method are excellent, and results compare well with those obtained by the Kageyama procedure [Clin. Chim. Acta 31, 421 (1971)].


Phospholipid composition of blood plasma, erythrocytes, and "ghosts" in sickle cell disease.

We examined the phospholipid composition of the plasma, of whole erythrocytes, and of "ghosts" of the erythrocytes from healthy volunteers, patients with sickle cell disease without crisis, and such patients in crisis, and found that phosphatidylglycerol in the plasma and "ghosts" was very significantly increased in sickle cell crisis but not in the absence of crisis. The significance of these findings is discussed.


Determination of acetaminophen concentrations in serum by high-pressure liquid chromatography.

We describe a method for determination of serum acetaminophen concentrations in serum by reversed phase high-pressure liquid chromatography. The homolog N-propionyl-p-aminophenol was used as an internal standard. The procedure, which requires only a single extraction with diethyl ether, can be optimized to be linear over the ranges of 10 to 100 or 1 to 20 mg/liter. Within-run CV was 1.2%; between-run CV was 4.4% and 4.9% at two different concentrations. Many commonly used drugs were tested and found not to interfere. The procedure is simple and rapid enough for use on an emergency basis in cases of overdosage, and can be optimized for measurement of either therapeutic or toxic concentrations.


Lysozyme determined in serum or urine by a simple nephelometric method.

We describe a nephelometric procedure for determination of lysozyme. A quantitative result is obtained in 2 to 3 min with 50 microliter of serum or urine. At a lysozyme concentration of 12.0 mg/liter, between-day precision was 3.4% and the analytical recovery 96.5-105.5%. The values obtained were a linear function of enzyme concentration below 50 mg/liter, and they agreed with those obtained by absorptiometric determination of the decrease in turbidity of a suspension of Micrococcus lysodeikticus.


Simultaneous radioassay of serum vitamin B12 and folic acid.

The radioassays of vitamin B12 and folic acid can be carried out in a single tube to give the simultaneous assay of both vitamins in 100 microliter of serum. Release of bound vitamins from their endogenous binders and the destruction of these binders are effected by a heating step at pH 9.3. The subsequent binding reactions with hog intrinsic factor and milk binder protein proceed advantageously and simultaneously in the same tube at pH 9.3. A single set of dual reagents replaces two sets of reagents that would normally be used for separate radioassays. Complete separation of bound radioactivities, [57Co]cyanocobalamin and 125I-labeled folate derivative, is obtained in a dual-channel gamma counter with no requirement for any correction for spill-over of counting data. Analytical results are comparable to those found for previously developed individual radioassays. The simultaneous assay has decreased technical time of analysis for these interrelated vitamins by about 50%.


Calculation of radioimmunoassay standard curves.

A method for calculating radioimmunoassay standard curves, based on the theory of Ekins et al., is described. Because a four-parameter model is used, nonlinear standard curves are the result. The calibration curve is fitted to the measured standard points by means of a weighted least-squares method. The program based on this model can be easily processed on a desk-top calculator. For all 250 runs of six different assays, very good standard curves could be obtained. The mean deviation between the concentrations of the standard points and the corresponding calculated values was about 6%. In 26% of the cases it could be shown that the model we describe gave significantly better results than did two simpler ones.


An automated competitive binding procedure for measuring thyroxine in serum.

A competitive binding assay for serum thyroxine has been automated, with the use of small, reusable Sephadex columns to separate thyroxine from endogenous thyroid-binding globulin and later to separate the bound and free thyroxine. Sixty samples an hour are run through columns, which are held in an aluminum turntable rotated by a fraction collector motor. Reagents and samples are fed to the columns by a proportioning pump. Waste eluates are collected and drained to the sink by a Teflon tray positioned between the columns and counting tubes, also held by the turntable. A cut out area in the tray allows one to collect the bound fraction, which can then be counted in a scintillation counter. Values are calculated and printed by a desk calculator interfaced with the scintillation counter. The "day-to-day" CV for this method is 5%. The accuracy is satisfactory when tested by comparison with other methods, by recoveries, and by linearity of dilutions.


Evaluation with the centrifugal fast analyzer of a chemical activation procedure for creatine kinase MB isoenzyme.

The differential activation method for the determination of the "myocardial" isoenzyme of creatine kinase (MB) is based on the computed difference in activity of serum aliquots activated by the combination of dithiothreitol and glutathione (skeletal plus myocardial creatine kinase) and by glutathione alone (skeletal creatine kinase). The unique ability of the Centrifugal Fast Analyzer to perform analyses in parallel is used to precisely measure (CV = 0.1-1.5%) the activity of the two chemically activated aliquots. Statistical considerations concerning the reliability of this difference estimation are discussed with respect to both the precision of the rate measurements and to the relative amount of isoenzyme MB present. Another potential source of error in the analysis of the two differently activated aliquots, namely variation in lag phases, is circumvented by use of a linear-search FOCAL software package. This program searches the data for a linear segment of maximum slope, automatically rejecting those data that appear in lag or depletion regions of the curve representing the progress of the reaction. Correspondence of the results obtained with those of comparison techniques (chromatography and electrophoresis) are discussed.


Determination of morphine by electron capture gas-liquid chromatography.

We describe a micromethod for determining as little as 0.1 microgram of morphine per liter of human urine. The procedure is about 20-fold more sensitive than are currently used gas-liquid chromatographic methods. It is particularly suitable for use as a confirmatory method for large-scale radioimmunoassay screening tests. The procedure involves extraction of morphine from urine and analysis of its heptafluorobutyryl derivative by gas-liquid chromatography with electron capture detection. Morphine can be determined by this procedure over a range of 0.1-200 microgram/liter of urine, 2 pg of the pure derivative being the lowest amount detectable. Urine samples from patients receiving methadone were analyzed by thin-layer chromatography, radioimmunoassay, and our procedure. The results by our procedure agreed well with those obtained by radioimmunoassay.


Simultaneous radioimmunoassay of thyrotropin and thyroxine in human serum.

The importance of early diagnois and treatment of congenital hypothyroidism has been well established, and several screening programs have been undertaken to detect neonates with this disorder by measurement of concentrations of thyrotropin or thyroxine in the serum. However, measurement of either hormone alone may fail to identify all affected patients. Accordingly, we have established a simultaneous double-antibody, dual-isotope radioimmunoassay for both. Sensitivity, slope, analytical recovery, and precision characteristics of the simultaneous assay do not differ from those of each assay performed separately. Values for the two analyses in the single and simultaneous assays correlate well (r = 0.951 for thyroxine, 0.983 for thyrotropin). This assay system permits determination of both hormones within 72 h after specimen collection and thus should allow more rapid evaluation, diagnosis, and treatment of infants with congenital hypothyroidism.


Detection of heterozygotes for phenylketonuria. Total body phenylalanine clearance and concentrations of phenylalanine and tyrosine in the plasms of fasting subjects compared.

Two tests have been compared for detection of heterozygotes for phenylketonuria, one based on determination of plasma phenylalanine and tyrosine concentrations in fasting individuals and the other on kinetic evaluation of the plasma elimination curve after intravenous loading with L-phenylalanine. The plasma elimination curve was biexponential and the kinetics were evaluated according to the two-compartment model. The constant, beta, expressing the rate of elimination from plasma at pseudo-equilibrium, the rate constant for the elimination from the central compartment, and the total body clearance were determined. Of these three, total body clearance, which on the average was reduced by 32% in the phenylketonuric heterozygotes, showed the best discriminatory ability, but was not better than the information on concentrations of phenylalanine and tyrosine in detecting heterozygotes for phenylketonuria.


Detection of heterozygotes for phenylketonuria by constant intravenous infusion of L-phenylalanine.

We measured the rate of elimination of phenylalanine by constant intravenous infusion of L-phenylalanine in 14 parents of children with phenylketonuria and in 21 subjects with a negative family history for this disease. When reciprocals of the observed elimination rates were plotted against the reciprocals of the increase in the plasma phenylalanine concentrations, approximately straight lines resulted. The theoretical maximum elimination rate, the mean value for which was 32 mmol/h in the reference subjects, was reduced by 41% in the phenylketonuric heterozygotes. The elimination rate at an increase in plasma phenylalanine concentration of 0.5 mmol/liter discriminated the phenylketonuric heterozygotes from normal homozygotes, with no overlap between the groups. A lower plasma tyrosine concentration in the phenylketonuric heterozygotes than in the reference subjects at the same rate of elimination of phenylalanine indicated an increased rate of elimination of tyrosine at a fixed concentration of this amino acid in these subjects.


Comparison of the Hoesch and the Watson-Schwartz tests for urinary porphobilinogen.

A comparison of the Hoesch and the Watson-Schwartz tests shows that the latter, although slightly more complicated, generally yields more concise results and is superior in sensitivity and specificity for porphobilinogen. The recommendation of the Hoesch test for use as a "bedside screening" method seems unrealistic. Before the diagnosis of an "inducible" porphyria is made, a positive Hoesch test requires that indoles, indoleacetic acid, methyldopa, end-stage alcoholic malnutrition, and phenazopyridine HCl be excluded, to avoid misinterpretation.


Determination of free thyroxine in serum by radioimmunoassay.

We describe a sensitive and specific radioimmunoassay for free thyroxine, directly determined in the diffusate obtained from equilibrium dialysis of serum. The intra- and interassay precisions were 7.5 +/- 0.27 and 9.1 +/- 0.58 ng/liter (mean +/- SD), respectively. The free thyroxine concentrations of normal persons and patients with thyroid disease were as follows: normal, 7.6 +/- 1.7; euthyroid patients receiving phenytoin, 6.9 +/- 1.4; hypothyroidism, 2.9 +/- 1.3; and hyperthyroidism, 32.3 +/- 12.3 ng/liter (mean +/- SD).


Plasma free amino acid patterns in pregnancy: relationships to gestational age and applications to detection of fetal distress.

Data on free amino acids in serial samples of plasma from 15 pregnancies from 14 women have been compared to data from normal nonpregnant women by use of pattern-recognition program. The overall amino acid patterns were found to be a reproducible function of gestational age. Two samples from a woman with a pregnancy terminated by a spontaneous abortion were clearly resolved from all the other samples by this procedure.


High-performance liquid chromatographic assay for 25-hydroxyvitamin D3 in serum.

We describe a precise and specific method for measuring 25-hydroxyvitamin D3 in 1 ml of human serum. Extraction with chloroform/methanol followed by chromatography on a 0.5 X 2-cm silica gel column yields a sample that is sufficiently free of extraneous material for high-performance liquid chromatography on a column of microporous silica gel (10 micron average particle diameter). The measurement is not influenced by vitamins D2 or D3, 25-hydroxyvitamin D2, or any of the more hydroxylated metabolites of the vitamin D group. Results by this method correlate well with a competitive protein-binding assay (r = 0.961), but with a negative bias of 6.9 +/- 3.3 microgram/liter. We measured concentrations of 25-hydroxyvitamin D3 in serum drawn during February from 24 persons who were judged normal by physical examinations. The range was 5.5-23.8 microgram/liter (mean, 15.0 +/- 5.2 microgram/liter). The day-to-day CV for the assay was 5.46%.


Simultaneously obtained skin-puncture serum, skin-puncture plasma, and venous serum compared, and effects of warming the skin before puncture.

Specimens of skin-puncture serum and plasma and venous serum were simultaneously obtained from healthy adults and in each specimen the concentrations of 12 chemical constituents were measured. No clinically important difference was found in the concentrations of the constituents measured in skin-puncture serum and plasma with or without warming the skin before puncture. When the concentration of each of the measured constituents was compared in skin-puncture specimens and venous serum there were important differences in the concentrations of glucose, potassium, total protein, and calcium. Except for glucose, the concentrations of these were higher in venous serum. The degree of hemolysis was the same in skin-puncture serum and plasma, but less in venous serum. The greater hemolysis in skin-puncture specimens was not reflected in a clinically important increase in the values of lactate dehydrogenase and potassium. We conclude that there is a difference in the concentration of some chemical constituents in skin-puncture specimens and venous serum and that these differences should be considered when results for these types of specimens are compared.


Urinary organic acids quantitated by age groups in a healthy pediatric population.

Urine samples from 92 individuals of a healthy pediatric population ranging in age from less than 24 h to 12 years were analyzed for organic acid content (amino acids excepted). A few samples from adults were included for purposes of comparison. Acids were extracted by using an anion-exchange procedure, converted to trimethylsilyloxime derivatives, and analyzed by gas chromatography with the help of a small computer system. Identifications were confirmed by using a combined gas chromatograph/mass spectrometer/computer system. The data has been tabulated according to five pediatric age groups. Despite large individual variation within age groups, significant differences in amounts of several acids excreted (based on urinary creatinine concentration) have been determined between age groups. Such differences might be due to one or a combination of several factors, including changes with age in creatinine excretion, in physiologic maturity, and in diet. Such data may help to provide a basis for diagnosing metabolic abnormalities in pediatric patients.


Ammonia production during clot retraction and its use in assay of fibrinoligase.

Clotting of recalcified plasma is followed by an increase in its ammonia content that lasts 4 to 6 h. This ammonia production closely parallels the increase in acid-insoluble fibrin, which is evidence that the ammonia results from the action of fibrinoligase. If the clot is removed, ammonia production stops. The initial velocity of ammonia production is directly proportional to the fibrinogen concentration in plasma. Thus the rate-limiting factor in normal shed blood is the fibrinogen concentration. A maximum of 6.4 +/- 1.5 (SD) molecules of ammonia are produced per molecule of fibrinogen. Determination of the total ammonia produced is the fastest direct method of estimating the extent of frbrin cross-linkage in whole plasma. A method is proposed for assaying fibrinoligase, based on the rate of ammonia production in the presence of casein as substrate. Normal values are 7.6 +/- 2.9 (SD) mumol/min per liter of plasma.


Analytical performance and comparability of the determination of cholesterol by 12 Lipid-Research Clinics.

Twelve Lipid-Research Clinic laboratories performed automated cholesterol analyses on four control-serum pools of known cholesterol concentration, using the Liebermann-Burchard reaction. The analyses were done during a two-year period, with the same standards, methodology, and quality-control procedures. Estimates of analytical bias, variability, and short- and long-term trends for each instrument and for the entire group of LRC instruments are presented. High accuracy, precision, and interlaboratory comparability were achieved through the rigorous standardization and control of the entire analytical procedure. The significance of these results for long-term collaborative studies is discussed. Individual laboratory biases averaged from 0.5 to 2.0% below Abell-Kendall reference values. Between-run variability was about equal to within-run variability and inter-laboratory variation was substantially less than intra-laboratory variation. The total standard deviation for all instruments was about 0.04 g/liter. Only 8-15% of this variation was due to differences between instruments. The between-instrument standard deviation ranged from 0.011 to 0.015 g/liter; the between-run, within-instrument standard deviation ranged from 0.023 to 0.030 g/liter; and within-run standard deviation ranged from 0.023 to 0.028 g/liter. The significance of the achieved results for long-term collaborative studies is discussed.


Serum alkaline phosphatase: normal values by sex and age.

We report here the normal range of serum alkaline phophatase activity as measured by the method proposed by Hausamen et al. [Clin. Chim. Acta 15, 241 (1967)] with a much larger sample size than used in previous investigations. In the statistical analysis the sample population is subdivided by sex and age, two variables which are known to influence enzyme activity. No statistically significant influence of blood type on enzyme activity was observed. The normal range of enzyme activity is reported in percentiles.


Effect of washing procedures on trace-element content of hair.

A pooled sample of hair was divided and portions prepared for analysis by three washing procedures, to evaluate the effect of washing procedure on the subsequent trace-element (Zn, Cu, Mg) content. The methods selected were a detergent wash, a hexane-ethanol wash, and an acetone-ether-detergent wash. For all elements, there was a significant difference among the results after these wash procedures. Magnesium content of hair was most affected by washing, containing less than half of the magnesium of the unwashed hair. The detergent wash removed the most zinc and magnesium; the acetone-ether-detergent wash removed the most copper. Our results indicate that the trace-element analysis of hair is sensitive to the preparation technique and therefore is an unreliable source of information about trace-element status.


Influence of methotrexate on results by three methods for determining protein in cerebrospinal fluid.

Because high intravenous and intrathecal doses of methotrexate are used in treatment for various neoplasms, this drug can reach high concentrations in cerebrospinal fluid. At those encountered clinically, it produced significant positive interference with values for cerebrospinal fluid protein as determined with the du Pont aca. A small interference was found with the Folin-Lowry method, and virtually no interference with an automated immunochemical procedure.


Measurement of serum and plasma ionic calcium with the "Space-Stat 20 Ionized Calcium Analyzer".

Ca2+ was measured in sera and plasma from 36 normal subjects (18 men 23-57 years old, and 18 women 20-52 years old) with the Space-Stat 20 Ionized Calcium Analyzer. Within-day and between-day precision (CV) for commercial serum-based specimens varied from 0.64-0.96% and 1.1-3.8%, respectively, depending on their mean concentrations. Plasma and serum showed within-day CV's of 1.05 and 0.79%, respectively. The linear regression of serum Ca2+ (y) on plasma Ca2+ (x) was y = 0.63x + 0.45 mmol/litre with a highly significant (P less than 0.001) coefficient of correlation (r) of 0.69. An approximate 5% increase in the mean Ca2+ concentration in serum over that found in plasma is probably due to heparin complexing. No significant sex-related difference was observed for either serum or plasma. The mean Ca2+ concentration of 29 refrigerated specimens tended to decrease in seven days (by 2.3%, not statistically significant). The normal Ca2+ range in serum and plasma (n = 35) was 1.08-1.18 and 1.02-1.14 mmol/litre, respectively.


Simultaneous assay for vitamin B12 and folate compared with separate assay.

Radioligand methods are now well established for measurement of serum vitamin B12 and folate. Often these two assays are requested together. Thus a way of doing them simultaneously would save considerable time and expense. The objectives of this study were to determine the precision of a simultaneous assay and to compare it with previously established assays for these vitamins.


Computer program for evaluation, physicochemical characterization and optimization of competitive protein binding assays: comparison of four curve-fitting models in peptide and steroid radioimmunoassays.

A modular construced computer program for evaluation, physicochemical characterization and optimization of compititive protein binding assays is put forward. The program allows standard curve fitting by the following models: spline approximation, semi-logarithmic interpolation, logit-log regression and logit-log interpolation. The suitability of these models was studied in a series of routine steroid radioimmunoassays (deoxycorticosterone, deoxycortisol, cortisol and aldosterone) and peptide radioimmunoassays (angiotensin I and angiotensin II). The spline approximation model was used as an approximate reference model. In all assays the logit-log interpolation model exhibited the best correlation with the spline approximation model. In the steroid assays the logit-log regression model was found to be inferior to the other models; in the peptide assays it was equal to or superior to the semi-logarithmic interpolation model. The study on the physicochemical parameters of the antigen-antibody reactions demonstrated that affinity of steroid antibodies was much lower than that of the peptide antibodies. Consequently the degree of antibody saturation at zero dose in the routine peptide assays was greater than 50% and in the routine steroid assays was less than 50%. The relationship of assay characteristics to the applicability of particular curve-fitting models is discussed.


Heme catabolism in human kidneys. Effect of various nephritides.

The role of the kidneys in heme metabolism was examined by determining heme oxygenase and biliverdin reductase activities in needle biopsy specimens from the kidneys of 11 female and 14 male patients with different renal diseases. The results show that both enzymes are present in human renal tissue. Our results indicate that heme oxygenase and biliverdin reductase increase slightly with age, but that there is no sex difference. A rising serum creatinine concentration is not reflected in the activity of heme degrading enzymes. Neither do the different types of nephritides, without detectable hemolysis, show significant differences in the renal activities of heme oxygenase and biliverdin reductase. This study thus outlines the basal physiological role of the human kidneys in heme degradation.


The differentiation of the porphyrias by means of high pressure liquid chromatography.

The analysis of faecal and urinary porphyrins by high pressure liquid chromatography (H.P.L.C.) provides characteristic profiles and facilitates rapid diagnosis of variegate (porphyria cutanea tarda hereditaria), symptomatic porphyria (porphyria cutanea tarda symptomatica), hereditary coproporphyria, acute intermittent porphyria, erythro-hepatic protoporphyria and congenital porphyria (erythropoietic porphyria).


Calcium levulinate medication. A pitfall in the diagnosis of organic acidurias.

Five children, who received calcium levulinate (calcium 4-oxopentanoate) intravenously in pharmacological doses excreted in the following 24-h period both 4-oxopentanoic acid (3.5--11.0 mg/24 h) and 4-hydroxypentanoic acid (4.5--10.4 mg/24 h). Attention is drawn to the fact that in gas chromatographic-mass spectrometric systems these compounds closely resemble the two acids found in children with beta-ketothiolase deficiency.


Relation of electrophoretic pattern of amylase isoenzymes to severity of pancreatic disease.

Serum and urinary amylase isoenzymes were studied in an attempt to evaluate the usefulness of isoamylase analysis in the diagnosis of pancreatic diseases. The amylase in human serum and urine was separated into 4 to 5 distinct isoenzymes, which have mobilities consistent with those of the pancreatic isoamylases (Amylase-1, -2, -4 and -6) and the salivary isoamylases (Amylase-3, -5 and -7). The normal isoamylase pattern was Amylase-1, -2, -3 and -5. Amylase-4 and -6 could not be found in normal serum and urine. Amylase activity in Amylase-1 and/or -2 increased remarkably, having the normal isoamylase pattern, in the patients with elevated serum amylase activity after stimulation with pancreozymin-secretin or after endoscopic retrograde cholangiopancreatography. However, in the patients with acute pancreatitis, salivary type isoamylases disappeared and only the pancreatic type components were identified. It seems that the appearance of Amylase-4 and disappearance of the salivary components indicate severe pancreatic inflammation and that the apparent specificity of this isoenzyme pattern affords a mean for conforming the diagnosis of acute pancreatitis. However, the occurrence of an inherited trait in the pancreatic isoamylases of healthy individuals suggests that a rise in the pancreatic components may not necessarily indicate pancreatic disorders.


Radioimmunoassay for subunit B in isoenzymes CK-MB and CK-BB of creatine phosphokinase.

We report the development of a double-antibody system for radioimmunoassay of CK-B subunit in isoenzymes CK-MB and CK-BB of creatine phosphokinase. With our method, 4.1 ng/ml of isoenzyme CK-BB can be detected. Within-run and between-run coefficients of variation are respectively 5.3% and 19.6% Analytical recovery of added antigen is 98.9 +/- 14.7%. The CK-B concentration in sera from 20 healthy adults was less than 75 ng/ml. In sera from 25 patients with a rise in CK-total activity, but without CK-MB or CK-BB activity tested with the previous reported immunological methods, we found CK-B concentrations in the range 0--86 ng/ml. In contrast, in 23 sera from patients with acute myocardial infarction within the last 48 hours CK-B concentration was in the range 104--225 ng/ml.


Electrophoretic characterization of pyrimidine 5'-nucleotidase of human erythrocytes and its distinction from acid phosphatase.

Electrophoresis of red cell pyrimidine 5'-nucleotidase was carried out on cellulose acetate strips. One major band of activity was found in preparations from human erythrocytes. This enzyme showed a specificity for the pyrimidine nucleotides UMP and CMP. No activity was detected with AMP. Pyrimidine 5'-nucleotidase activity could be separated from that of acid phosphatase with the use of alpha-glycerophosphate. This method may be useful in the study of pyrimidine 5'-nucleotidase deficiency in red cells.


Bile 5'-nucleotidase in the serum of jaundiced rats.

Rat serum 5'-nucleotidase, L-leucyl-beta-naphthylamidase and beta-glycerophosphatase activities are increased whilst alkaline p-nitrophenylphosphatase and alkaline phosphodiesterase activities are unchanged or decreased three days after bile duct ligation. Affinity chromatography on an immobilised antiserum raised against highly purified liver plasma membranes showed that although 5'-nucleotidase in normal serum is unrelated to the 5'-nucleotidase of liver plasma membrane, the 5'-nucleotidase of bile and much of the 5'-nucleotidase in the jaundiced serum are closely related to the plasma membrane enzyme. Since bile is rich in 5'-nucleotidase, the changes in level of this enzyme after bile duct ligation are most simply explained by leakage of bile into the blood; changes in the patterns of the other enzymes are shown to be consistent with this explanation. The jaundiced serum was examined by gel exclusion chromatography and flotation in sucrose gradients for the presence of small fragments of plasma membrane as reported in human jaundiced sera, but no such fragments could be detected three days after bile-duct ligation.


Quantitative determination of 5-hydroxyindole-3-acetic acid in body fluids by high-performance liquid chromatography.

A method for the quantitative determination of 5-hydroxyindole-3-acetic acid in urine and cerebrospinal fluid is described. The method is based on reversed phase high-performance liquid chromatography and fluorometric detection. The experimental error was below 5% in urine and 8% in cerebrospinal fluid. Urine and cerebrospinal fluid samples were analysed both by the described method and by a mass fragmentographic method. The correlation coefficients for the two methods were 0.986 in the urine samples and 0.965 in the cerebrospinal fluid samples. Analysis of 100 samples can be achieved within 3 days. The described method offers a possibility to determine 5-hydroxyindole-3-acetic acid in urine and cerebrospinal fluid with the same precision and capacity as do mass fragmentography but with the use of a less sophisticated equipment.


Lactate dehydrogenase isoenzymes of human erythrocyte membranes.

Human erythrocyte membranes and cytosol were fractionated in detergent-containing polyacrylamide gels and stained for lactate dehydrogenase activity. A highly significant difference in the distribution of three isoenzymes of lactate dehydrogenase (LD-1, LD-2 and LD-3) between membranes and cytosol was found, a striking feature being the poor binding of LD-3 to the membrane fraction.


Relationship between carbamazepine concentrations in plasma and saliva in man as determined by liquid chromatography.

The concentration of carbamazepine in plasma and saliva was determined in 7 subjects receiving carbamazepine. Plasma and saliva samples were assayed was an excellent linear relationship between carbamazepine concentrations in saliva and plasma (r = 0.991, p less than 0.001) over a plasma concentration range of 0.2--8.0 microgram/ml. The saliva/plasma ratio for carbamazepine was 0.26 +/- 0.02 (S.D.). Since carbamazepine binding to plasma proteins is in the order of 75%, the saliva concentration seems to reflect the concentration of the free drug in plasma. There is very little intrasubject and intersubject variation in the concentration ratio of carbamazepine. This study demonstrates that carbamazepine determination in saliva is a convenient method for optimizing blood levels of patients as well as for the study of pharmacokinetic properties.


Reduced level of glycine cleavage system in the liver of hyperglycinemia patients.

The activity of the glycine cleavage system in liver of hyperglycinemia with organic acidemia was 1/6 to 1/20 tha of normal human livers. The reduced activity results from the reduction in concentration of the enzyme of the glycine cleavage system. All the protein components of the glycine cleavage system examined were reduced when the activity was determined seprately. H-protein was purified from patients' and control livers, and there was found no difference in their chromatographic properties.


Evaluation of a microspectrofluorimetric method or serum vitamin A.

A microspectrofluorimetric method for vitamin A was compared with spectrophotometric and trifluoroacetic acid-colorimetric procedures. Values obtained by the fluorimetric method showed a high degree of correlation with those obtained by other standard procedures. However, it was found that the fluorimetric method yielded values which were several-fold higher than those obtained with the spectrophotometric method when sera from pregnant women were tested. The fluorescence from sera of pregnant subjects showed a triphasic decline even when the samples were stored at --20 degrees C.


Homocysteine and cysteine loads in patients with homocystinuria due to cystathionine synthase deficiency: effects of vitamin B-6.

The metabolic response of patients with homocystinuria due to cystabhionine synthase deficiency to oral loads of homocysteine indicates: that even severely affected patients with homocystinuria have pools of cystine in their tissues; that control of sulfur amino acid metabolism favors increased concentrations of methionine rather than homocystine in the plasma; and that even patients who apparently are not B-6-responsive respond differently to the loads of homocysteine when challenged during B-6-treatment compared with their response before B-6 treatment. Loading tests with homocysteine indicate that B-6 treatment be of some benefit even in individuals who do not have an obvious biochemical response to such therapy.