Allan-Herndon syndrome. I. Clinical studies.

A large family with X-linked mental retardation, originally reported in 1944 by Allan, Herndon, and Dudley, has been reinvestigated. Twenty-nine males have been affected in seven generations. Clinical features include severe mental retardation, dysarthria, ataxia, athetoid movements, muscle hypoplasia, and spastic paraplegia with hyperreflexia, clonus, and Babinski reflexes. The facies appear elongated with normal head circumference, bitemporal narrowing, and large, simple ears. Contractures develop at both small and large joint. Statural growth is normal and macroorchidism does not occur. Longevity is not impaired. High-resolution chromosomes, serum creatine kinase, and amino acids are normal. This condition, termed the Allan-Herndon syndrome, appears distinct from other X-linked disorders having mental retardation, muscle hypoplasia, and spastic paraplegia.     

THE ABPHYL SYNDROME IN ZEA MAYS. I. ARRANGEMENT. NUMBER AND SIZE OF LEAVES

A range of phyllotactic variation in Zea mays has been obtained from progeny derived from a single ‘opposite-leaved’ plant. Some segregants exhibit a form of alternate phyllotaxy with poor separation between nodes, while others duplicate the original decussate condition. Numerous intermediate examples have also been observed. Some have both spiral and opposite arrangements on the same plant; others, which begin their ontogeny with the normal distichous arrangement, switch at different stages of maturity to spiral or decussate arrangement. Leaves from ABPHYL plants are up to one-half the width of comparable normal leaves although their length is similar. Since ABPHYL plants may have twice as many leaves as do normal siblings, total area and dry weight of leaf blades per ABPHYL plant are greater than in normal siblings. Leaf width of both ABPHYL and normal plants correlates well with the number rather than the width of epidermal cells.     

Beta-glycosidases and diabetic microangiopathy. II. An insulin-dependent isozyme of beta-N-acetylglucosaminidase.

Previously we reported that beta-glycosidase activities were markedly decreased in the kidney but increased in the serum of diabetic rats. To examine these changes, the isozymes of beta-N-acetylglucosaminidase [EC 3.2.1.30] of rats were examined by DEAE-cellulose column chromatography. At least 3 major isozymes were found in both the kidney and liver. The main isozyme was type II isozyme in normal rat kidney and type III in normal rat liver. The activity of the type II isozyme in the kidney was markedly lowered when the total activity was decreased in diabetes and its normal activity was restored on insulin treatment, in parallel with increase in the total activity in diabetes. No significant change was found in the chromatographic pattern of isozymes in the liver in diabetes. In diabetic rat serum, the increase of total activity was found to be due to increase of type I and II isozymes.     

Bloom's syndrome. IV. Sister-chromatid exchanges in lymphocytes.

An abnormally great amount of exhange between both sister and nonsister-but-homologous chromatids is a highly characteristic feature of cultured blood lymphocytes from individuals with Bloom's syndrome. However, a population of lymphocytes which exhibit a normal amount of exchange can be detected in the blood of some individuals with this syndrome. This coexistence of cells with a greatly increased number of sister-chromatid exchanges and others with a normal number results in a phenotypic dimorphism, in apparent contradiction to the autosomal recessive mode of inheritance of the syndrome.     

Normal and benign human prostatic epithelium in culture. I. Isolation.

Isolation of normal human glandular epithelia and their growth and maintenance in vitro have been major problems. The primary objective of studies presented here was to isolate postpubertal, normal human, viable prostatic epithelium for in vitro cultivation. The long-term objective of these investigations was to develop an in vitro human cell model system for studies on prostatic carcinogenesis. A method for isolation of viable, normal and benign human prostatic epithelium, using collagenase for tissue dissociation, is described. Intact acini were isolated, which, on plating gave rise to vigorously growing monolayer cultures of epithelial cells. The purity of epithelial cultures partly depended upon the source of tissue. Specimens of normal prostate and those of benign tissue derived from open prostatectomies provided primarily pure epithelial cultures with occasional fibroblast colonies in some cultures, which could be removed. Cultures from some specimens of transurethral resection of the prostate (TURP) contained many fibroblast colonies due to incomplete separation of acini from the stroma. This resulted from incomplete digestion of denatured tissue caused by electrocauterization during surgery. Cultures established in this manner are being used to study the effects of hormones, vitamins and other growth regulators in order to establish growth requirements of these cells in vitro, which would facilitate their long-term maintenance.     

Antigen modulation of the secondary immune response. VI. Contribution of cells recruited from precursor pool to expression of memory.

The adoptive transfer system has been used extensively to study the ability of antigen triggered memory cells to become antibody forming cells and/or to proliferate and expand the memory cell population. Selective antigen triggering of the memory cells for low and high affinity antibody formation has also been studied in this way. One of the main counter-arguments to the interpretation of these data is that the presence of antigen in the adoptive host may lead to recruitment of new memory cells from either a host or donor precursor population. In this paper we examined the contribution of both host and donor precursor cells to the total antibody response in adoptive secondary recipients. The following donor-host combinations were used in which the recipients were given 1 mg fluid antigen intravenously: (A) normal (non-immune) donors to normal irradiated recipients; (B) normal donors to carrier primed irradiated recipients; (C) carrier primed donors to normal irradiated recipients; (D) normal donors to carrier primed recipients with challenge and subsequent transfer to additional carrier primed recipients; (E) carrier primed donor to normal recipients to carrier primed recipients; (F) repeat of B and C above with multiple antigen administration; (G) purified immune (DNP-BGG) donor T cells mixed with normal B cells transferred to normal irradiated recipients. In most cases recruitment was seen but this represented less than 4% of the responses seen with immune cells. Thus we conclude that this level of recruitment does not compromise the use of the adoptive transfer system for studying selective antigen triggering of memory cells.     

Driving activity. A quantitative study.

A comparative study of driving activity between normal subjects and neurological patients was performed. Driving activity was considered as the energy of the visual evoked potentials filtered at the same frequency of stimulation (1, 2, 4, 7, 10, 12 and 15 cps) using a CAT 400 C computer as a digital filter. The hemispheric symmetry of the responses was measured by the Pearson product moment correlation coefficient and the signal energy ratio. Each symmetry measure for every patient was compared with the normal values and considered abnormal when differences were greater than 3 SD from the normal mean. Of 25 patients 14 of them with a normal EEG, 23 presented severe alterations in the symmetry of the filtered visual evoked responses. Each patient showed a peculiar pattern of abnormality. It is concluded that the procedure described is a very powerful method in the discrimination of brain lesions.     

Lipoproteins in lecithin-cholesterol-acyltransferase(LCAT)-deficiency. II. Further studies on the abnormal high-density-lipoproteins.

The lipoproteins from two sibs with familial lecithin-cholesterol-acyltransferase(LCAT)-deficiency were further characterized. Comparatively lipoproteins from patients with secondary LCAT-deficiency were studied. Both groups of patients had particles of unusual size and shape in the alpha1-(HD-2)-lipoprotein subfraction. The abnormal HDL-2 particles were disk-like in appearance with a major axis of about 180 A and a minor axis of about 40 A and tended to aggregate into long coinlike stacks. The abnormal HDL-2 particles contained the normal protein constituents of HDL Apo A-I, Apo A-II and Apo C but in addition a major polypeptide with a M.W. of 39000 not seen in significant amounts in normal high-density-lipoproteins. This polypeptide was found identical in size, isoelectric focusing and immunochemically with an arginine-rich normal polypeptide constituent of very-low-density-lipoproteins designated apoprotein E. Presence of this protein marker in the HDL allowed the specific immunological detection of the abnormal HDL-2 (LP-E) in plasma. Further minor biochemical abnormalities were observed in the lipoproteins of the patients with familial LCAT-deficiency. However, the main protein constituents of their HDL, the Apo A, Apo C and Apo E polypeptides, were found to be identical electrophoretically and by analytical isoelectric focusing with their normal counterparts. The data suggest that the basic genetic defect in the hereditary disease leads to a deficient activity of the LCAT-enzyme and that all abnormalities in the lipoprotein spectrum are secondary.     

Combined deficiency of factor V and factor VIII. A report of another case.

A patient with combined factor V and factor VIII deficiency is presented. The bleeding manifestations were: easy bruising, post-traumatic bleeding, bleeding after tooth extractions. The main laboratory feature was a prolonged partial thromboplastin time which was corrected by the addition of adsorbed normal plasma but not by the addition of normal serum, hemophilia A plasma of another patient with combined factor V and factor VIII deficiency. The thromboplastin generation test was clearly abnormal and was corrected by the addition of adsorbed normal plasma but not by addition of normal serum. Prothrombin consumption was also defective. Prothrombin time was slightly prolonged too, Thrombin time, platelet and vascular tests were within normal limits and there was no hyperfibrinolysis. Factor VIII was 8% of normal, whereas factor V was 14% of normal. Factor VIII associated antigen was normal. All other clotting factors were within normal limits. The parents of the propositus were consanguineous (first cousins) but had normal factor V and factor VIII activity and normal factor VIII antigen. The same was true for other family members. The hereditary transmission of the condition appears autosomal recessive.     

Membrane function in cystic fibrosis. I. Putrescine transport in normal and cystic fibrosis fibroblasts

Putrescine transport was examined in normal and cystic fibrosis fibroblasts. No differences were observed in accumulation pattern, kinetics of uptake, or efflux between CF and normal cells. In both growing and growth-arrested CF and normal fibroblasts, exogenously supplied putrescine remained unchanged for at least 60 min. Some differences were observed in the response of CF and normal cells to environmental (media) changes.This research was supported by a grant from the Cystic Fibrosis Foundation and by a grant from the National Institutes of Health, Training Grant (GM01316 11 GNC).     

Normal mode refinement: crystallographic refinement of protein dynamic structure. I. Theory and test by simulated diffraction data.

A dynamic structure refinement method for X-ray crystallography, referred to as the normal mode refinement, is proposed. The Debye-Waller factor is expanded in terms of the low-frequency normal modes whose amplitudes and eigenvectors are experimentally optimized in the process of the crystallographic refinement. In this model, the atomic fluctuations are treated as anisotropic and concerted. The normal modes of the external motion (TLS model) are also introduced to cover the factors other than the internal fluctuations, such as the lattice disorder and diffusion. A program for the normal mode refinement (NM-REF) has been developed. The method has first been tested against simulated diffraction data for human lysozyme calculated by a Monte Carlo simulation. Applications of the method have demonstrated that the normal mode refinement has: (1) improved the fitting to the diffraction data, even with fewer adjustable parameters; (2) distinguished internal fluctuations from external ones; (3) determined anisotropic thermal factors; and (4) identified concerted fluctuations in the protein molecule.     

The reliability of eye movement quantitative evaluation. I. Saccadic eye movements.

We adopted the estimate of the intraclass coefficient of reliability, R, to evaluate the reliability of saccadic eye movements quantitative analysis. At a one-week interval we recorded refixative saccadic eye movements twice from fifteen healthy subjects by means of the binocular electrooculographic technique. R was computed for the constants and the slopes of the amplitude/duration and the amplitude/peak velocity relationships, for the mean precision values and for the mean latency values adjusted for subject's age. Our data demonstrated that the reliability of saccade parameters is fairly good for the amplitude/peak velocity relationship, good for the precision and very good for the amplitude/duration relationship. Finally, we believe that the normal variability values we obtained can be usefully employed in neurophysiological longitudinal studies not only in normal subjects, but also in pathological condition provided the more reliable parameters and the more adequate strategies to compute normal variability values are chosen.     

Time domain dielectric spectroscopy study of human cells. II. Normal and malignant white blood cells.

The dielectric properties of human lymphocyte suspensions were studied by time domain dielectric spectroscopy (TDDS). Nine populations of malignant and normal lymphocytes were investigated. Analysis of the dielectric parameters of cell structural parts were performed in the framework of Maxwell-Wagner mixture formula and the double-shell model of cell. The specific capacitance of the cell membranes was estimated by the Hanai-Asami-Koisumi formula. It was shown that the dielectric permittivity, capacitance and conductivity values of cell membranes are higher for normal lymphocytes than for the malignant ones. The difference of the same parameters for normal B- and T-cells is also discussed.     

Os penis of the rat. V. The distal cartilage process

The morphogenesis and morphology of the distally positioned cartilage of the os penis, the processus cartilagineus, are described in rats aged from 1 to 100 days. Based on observations of metachromacy of the process stained with toluidine blue it was found that a processus cartilagineus only exists in the period between 35 and 50 days of age. Before 35 days, the structure consists of connective tissue proper, and after 50 days the cartilage starts to calcify partially. The present paper also initiates studies of experimentally caused alterations of the normal development of the processus cartilagineus by subjecting 35-day-old rats to castration, with subsequent sacrifice at 100 days. Castration at that age causes a complete interruption of normal development of the processus cartilagineus as the structure in 100-day-old castrated rats has distinct morphological characteristics in common with those of 35-day-old normal rats. The present paper, thus, confirms that normal development of the processus cartilagineus seems to be male-hormone-dependent.     

Lipid metabolism in cultured cells. XVI. Lipoprotein binding and HMG CoA reductase levels in normal and tumor virus-transformed human fibroblasts.

The loss in feedback control of cholesterol biosynthesis in tumor cells was examined in tissue culture. Human fibroblasts from normal subjects, SV40 tumor virus-transformed cell lines, and homozygous familial hypercholesterolemic cells as reference, were grown in tissue culture. Experiments were conducted to relate the regulatory enzyme for cholesterol biosynthesis, HMG CoA reductase, and the membrane-located binding receptors for low density lipoproteins (LDL) that mediate feedback control in normal cells. Monolayers of virus-transformed tumor cells exhibited specific (125)I-labeled LDL binding of 152 +/- 21 ng/mg cell protein, which was essentially the same as that of normal fibroblasts (135 +/- 20 ng/mg). Binding of LDL by familial hypercholesterolemic cells used as controls was only 8 +/- 3 ng/mg under the same test conditions. Basal levels of HMG CoA reductase in tumor cells of 45.2 +/- 6.5 units/mg cell protein were about twice those of normal cells. However, in contrast to the lack of feedback control of this enzyme observed with tumors in vivo, in both the normal and the transformed cells in vitro, activity of the enzyme decreased about fourfold when serum lipids were added. These findings demonstrate that tumor cells growing in vitro contain a normal complement of the membrane-located binding receptors for low density lipoproteins and, although the basal levels are higher than normal, an effective feedback regulation of the enzyme HMG CoA reductase is retained.     

Canine cardiac myosin with special referrence to pressure overload cardiac hypertrophy. I. Subunit composition.

In studies of myosin from left and right ventricles of normal hearts and hypertrophic hearts at 5 weeks and 13 weeks after aortic banding, polyacrylamide gel electrophoresis shows intermediate molecular weight components which derive from heavy chains fragmented in the presence of dodecyl sulfate. The proportion of degraded heavy chains is greater in myosin from hypertrophic hearts than normal hearts, with comparable degradation in left and right ventricle myosin. The observed fragmentation of myosin results from proteolysis due to contaminant proteases or a thermally activated, heat-stable nonenzymatic process, or both. The susceptibility of heavy chains to crude myofibrillar proteases differs in normal and hypertrophic cardiac myosin; however, the kinetics of tryptic digestion are identical for both myosins. With precautions to minimize proteolytic artifacts on dodecyl sulfate-polyacrylamide gel electrophoresis, preparations of myosin from left and right ventricles of normal and hypertrophic hearts exhibit comparable subunit composition, with approximately molar ratios of heavy chains, light chain L1, and light chain L2. Comparable stoichiometry for the light chain fraction is determined by high speed sedimentation equilibrium at pH 11 and direct fractionation of the different cardiac myosins. We do not confirm reports (e.g. Wikman-Coffelt, J., Fenner, C., Smith, A., and Mason, D. T. (1975) J. Biol. Chem. 250, 1257-1262) of different proportions of light chains in left and right ventricle myosin of normal and hypertrophic canine hearts. The light chains display microheterogeneity, with L1 generating two isoelectric variants and L2 generating two major and two minor variants, but identical mobilities and isoelectric values are obtained in the different myosin preparations.     

NMR in cancer. VII. Sodium-23 magnetic resonance of normal and cancerous tissues.

Sodium-23 magnetic resonance was performed on four types of cancers and six types of normal tissues of rats and mice. The spin-lattice relaxation time of the tumors was generally longer than that of the normal tissues, with the most marked difference occurring between rat liver (T1 = 6.5 msec) and Novikoff hepatoma (T1 =23.7 msec). Estimation of tissue sodium from the signal intensity of the resonance indicated that all four types of tumors contained more sodium than any of the normal tissues.     

Human colonic adenocarcinoma cells. I. Establishment and description of a new line.

A series of human colonic epithelial cell lines have been cultured from a single patient: LS-180 the original adenocarcinoma, LS-174T a trypsinized variant, and normal colonic tissue. The malignant cells, 20 to 40, mum in diameter and oval to polygonal, exhibited characteristics of normal colonic mucosal cells, namely, abundant microvilli prominent in secretory cells, and the presence of intracytoplasmic mucin vacuoles. The cultured adenocarcinoma cells, but not normal, demonstrated neoplastic properties by producing high levels of carcinoembryonic antigen (CEA) and by the ability to be propagated in hamster cheek pouches and in immunodeprived mice. The CEA production by the newly established line LS-180 released 900 times more CEA per cell into the culture medium and bore 30 times more cell-associated material than the established line, HT-29. These cell lines may permit detection of distinctive chemical, physiological, pharmacologic, and immunologic characteristics of neoplastic colonic cells.     

Androgen metabolism by rat epididymis. 3. Effect of castration and anti-androgens.

The in vivo and in vitro metabolism of 3H-testosterone by rat epididymis and the changes in epididymal weight have been studied after castration and treatment with anti-androgens. The utilization of 3H-testosterone was greatly reduced after castration as was the formation of 5alpha-reduced 17 beta-hydroxy metabolites. The formation of the 17 -keto metabolites was unaffected. Castration had no effect on the ratio between water and ether soluble radioactivity. Administration of testosterone propionate, necessary for giving normal stimulated prostate weight (150 mug/day), restored the metabolism of testosterone to approximately normal values. Estradiol benzoate and progesterone inhibited metabolism of testosterone in vitro and greatly reduced the formation of DHT (17 beta-hydroxy-5alpha-androstan-3-one) and 3 alpha-diol(5 alpha-androstane-3 alpha-17 beta-diol) by experiments both in vivo and in vitro. No effect of cyproterone acetate could be demonstrated on either the in vitro or in vivo metabolism of testosterone. Castration for 14 days reduced the epididymal weight to about 30% of that found in intact animals. Administration of testosterone propionate restored the epididymal weight to about 80% of normal. Estradiol benzoate and cyproterone acetate given to intact rats led to a decrease in the epididymal weight. Progesterone had no such effect. In 14 days castrated rats receiving testosterone propionate all three anti-androgens reduced the weight of the epididymis. In conclusion, our results show that the metabolic conversion of testosterone in epididymis to DHT and 3 alpha-diol is dramatically dependent on the hormonal status of the animal; castration or treatment with anti-androgens causes a reduced formation of the "active" androgens whilst testosterone replacement treatment restores the metabolism of testosterone to normal.     

Resistance to Suppression by Dexamethasone of Plasma 11-O.H.C.S. Levels in Severe Depressive Illness

Use of the midnight dexamethasone suppression test showed that the plasma 11-hydroxycorticosteroid (11-O.H.C.S.) level did not undergo its normal reduction in 14 out of 27 patients with severe depression. Resistance to dexamethasone suppression correlated with the clinical rating of the severity of depression, while recovery from depression was associated with return of normal responsiveness to dexamethasone. The explanation of these findings is unknown.