Deoxyribonucleic acid replication in human diploid fibroblasts stimulated to proliferate.

The elongation of replicating DNA chains was studied in resting (G0) cultures of WI-38 cells stimulated to proliferate. An alkaline sucrose gradient sedimentation method which avoids shear effects was employed. The earliest detected intermediate is a short (4.5 S) piece of DNA which is converted to 11 S and larger (50 to 60 S) pieces within 1 min. The 50 to 60 S pieces are gradually converted to two distinct size classes, 212 S and 275 S, which are in turn converted to a chromosomal aggregate of about 400 S. The overall rate of elongation of DNA is about 1.4 X 10(6) daltons/min.


Studies on Limulus amoebocyte lysate II. Purification of the coagulogen and the mechanism of clotting.

A coagulogen in Limulus lysate has been purified to apparent homogeneity as judged by sodium dodecyl sulfate-gel electrophoresis. The purified coagulogen identified by its ability to clot with either trypsin or the purified Limulus clotting enzyme (Tai, J.Y., and Liu, T. Y. (1976) Fed. Proc. 35, 1486) has a molecular weight of 24,500. It consists of a single polypeptide chain of about 220 amino acids with glycine and serine as its NH2- and COOH-terminal residues; respectively. When acted upon by the Limulus clotting enzyme, the coagulogen releases a soluble C-peptide of about 45 amino acids and an insoluble coagulin of about 170 amino acids. The latter interacts in a noncovalent fashion to form the clot. Amino acid analyses together with the results of NH2- and COOH-terminal analyses suggest that the clot formation involves the cleavage of an--Arg--Lys--bond. Trypsin acts on the coagulogen to cause clotting by splitting the same peptide bond.


Re-evaluation of the kinetic mechanism of the choline acetyltransferase reaction.

Initial velocity patterns for human placental choline acetyltransferase show a series of converging lines for both the forward and reverse reaction; with Kacetyl coenzyme A = 11.9 micron, Kcholine = 0.41 mM, KCOA = 8.8 micron, and Kacetylcholine = 1.3 mM. The relative rates of acetylcholine synthesis (Vf) to acetylcholine breakdown (Vr) is 4.5. Product inhibition by acetylcholine is competitive with respect to choline and noncompetitive with respect to acetyl-CoA, while product inhibition by choline is competitive with respect to acetylcholine and noncompetitive with respect to coenzyme A. Chlorocholine, diethylaminoethanol, and acetylaminocholine were used as dead-end inhibitors and shown to inhibit competitively with respect to acetylcholine, and noncompetitively with respect to choline, acetyl-CoA, and CoA. At high choline concentrations, uncompetitive substrate inhibition is observed, and inhibition by acetylaminocholine changes from noncompetitive to competitive. Comparing the reactivity of dimethylaminoethanol to choline, and acetyldimethylaminoethanol to acetylcholine, the maximal velocities obtained with these analogues was approximately 25% of the natural substrates. These data are not consistent with the previously proposed ordered Theorell-Chance reaction mechanism, and have been interpreted in terms of a random binding mechanism.


Exchange of cytochrome b5 between phospholipid vesicles.

Detergent-extracted cytochrome b5, which binds readily to phosphatidylcholine vesicles, has been shown to exchange between vesicles by the following experiments. When preformed cytochrome b5-vesicle complexes are incubated with fresh vesicles a new species arises which sediments in the analytical ultracentrifuge with a rate intermediate between those of the original components. When the same mixture is subjected to free boundary electrophoresis no slow moving component corresponding to pure vesicles is seen. The results of sucrose density gradient centrifugation are also consistent with the conclusion that cytochrome b5 exchanges between vesicles. The possibility that the above results were produced by vesicle fusion or aggregation was eliminated by light-scattering studies and a direct assay for vesicle fusion. The ability of cytochrome b5 to exchange between lipid structures has important implications not only in the cytochrome b5-lipid system but also in enzymatic reactions in which cytochrome b5 is involved.


Amino acid sequence of phospholipase A2-alpha from the venom of Crotalus adamanteus. A new classification of phospholipases A2 based upon structural determinants.

The complete amino acid sequence of Crotalus adamanteus venom phospholipase A2-alpha has been determined by analysis of the five tryptic peptides from the citraconylated, reduced, and S-[14C]carboxamidomethylated enzyme. Earlier studies (Tsao, F. H. C., Keim, P. S., and Heinrikson, R. L. (1975) Arch. Biochem. Biophys. 167, 706) provided the information necessary to align the tryptic fragments so that secondary cleavage procedures to establish overlaps were unnecessary. The subunit in the phospholipase A2-alpha dimer is a single polypeptide chain containing 122 amino acids and seven disulfide bonds. The histidine residue implicated in the active site of mammalian phospholipases is at position 47 in the C. adamanteus enzyme and is located in a domain of the molecule which is highly homologous in sequence with corresponding regions of phospholipases from a variety of venom and pancreatic sources. Comparative sequence analysis has revealed insights with regard to the function and evolution of phospholipases A2. Primary structural relationships observed among the snake venom enzymes parallel the phylogenetic classification of the venomous reptiles from which they were derived. It is proposed that phospholipases A2 of this general type be divided into two groups depending upon the presence or absence of distinctive structural features elucidated in this study.


Human beta-globin messenger RNA. I. Nucleotide sequences derived from complementary RNA.

Sequence analysis studies were carried out on human beta-globin mRNA (beta-mRNA) prepared from alpha-thalassemic, sickle cell, and Hb A reticulocytes. Highly purified beta-mRNA served as substrate for the preparation of cDNA by RNA-dependent DNA polymerase. The cDNA was transcribed by Escherichia coli RNA polymerase and the resulting cRNA was analyzed. Over 300 nucleotides were assigned to the beta-mRNA coding region and 37 nucleotides were assigned to the 3'-terminal noncoding region. The normal termination codon is UAA which is separated by 28 nucleotides from an out of phase UAA triplet. The origin of each of the abnormally long beta-globin variants Tak and Cranston is consistent with reduplication of dinucleotides prior to the normal termination codon, and both globin variants can terminate at the out of phase UAA.


The purine nucleotide cycle. Studies of ammonia production by skeletal muscle in situ and in perfused preparations.

Ammonia production by hind limbs of rat was studied in perfused preparations during rest, exercise, and hypoxia, and while the perfusate contained epinephrine or cyanide. Ammonia production by hind limbs was also studied in situ during rest and exercise. Strong correlations were observed between ammonia, IMP, and lactate formation. Adenylosuccinate was not present in detectable amounts in resting muscle (less than 1 nmol/g dry weight) but it appeared and rose sharply in exercising muscle, and then declined during recovery. The maximum found was about 18 nmol/g dry weight. When IMP accumulation exceeded 2.8 mumol/g dry weight, the formation of adenylosuccinate diminished, presumably because of an inhibition of adenylosuccinate synthetase by excess substrate. Exercise resulted in a greatly increased output of ammonia and a decreased output of alanine and glutamine. The tissue contents of aspartate and glutamate were decreased, while that of alanine was increased. The content of malate increased 3-fold during exercise. Epinephrine, hypoxia, and cyanide each caused an increased output of ammonia at the expense of the amino acid content of the tissue. These results provide evidence for the operation of the purine nucleotide cycle in skeletal muscle under various conditions that are associated with an increased rate of glycolysis.


Hepatic microsomal dealkylations. Inhibition by a tyrosine-copper (II) complex provided with superoxide dismutase activity.

The effect of a divalent copper-tyrosine complex has been evaluated in rat liver microsome-catalyzed dealkylations. The copper complex, which is provided with superoxide dismutase activity, inhibits at micromolar concentrations aminopyrine, p-nitroanisol, and 7-ethoxycoumarin dealkylations. It has also been found that cumene hydroperoxide-supported p-nitroanisol demethylation, the formation of a 440 nm species, and the formation of superoxide radicals are inhibited by the divalent copper complex. On the other hand, 3-chloroperbenzoic acid has been found to support a copper complex-insensitive 7-ethoxycoumarin dealkylation. Oxygen uptake by rat liver microsomes is also inhibited by the copper complex. The data support the concept that the copper complex acts as a superoxide dismutase at the level of a cytochrome P-450 intermediate species, liganded with superoxide anions.


Steady state kinetics of 4-hydroxyphenylpyruvate dioxygenase from human liver (III).

Kinetic experiments have been made with an apparently homogenous preparation of human liver 4-hydroxyphenylpyruvate dioxygenase Form 3 (4-hydroxyphenylpyruvate: oxygen oxidoreductase (hydroxylating, decarboxylating), EC 1.13.11.27) at 37 degrees in 0.2 M Tris/HCL, pH 7.5, by measuring the evolved carbon dioxide from the 1-14C-labeled substrate or the formation of homogentisate from the U-14C-labeled substrate. The effect of variations in the concentrations of substrates, products, and metal chelators on the velocity of the forward reaction was studied. The results agree with an Ordered Bi Bi kinetic mechanism (Cleland, W. W. (1963) Biochim. Biophys. Acta 67, 104--137), where 4-hydroxyphenylpyruvate is added prior to oxygen and CO2 released before homogentisate. A Theorell-Chance mechanism has not been excluded.


Membrane polypeptides co-induced with the bacterial bioluminescent system.

Electrophoretic analysis of membrane proteins and electron microscopy of freeze-fracture replicas reveal that certain polypeptides and intramembrane particles are absent in Beneckea harveyi harvested prior to luminescence induction but present after induction. The polypeptides and particles are also absent in mutants which fail to synthesize the luminescent system. These correlations implicate membrane polypeptides in the bacterial bioluminescent system.


Blood compatibility of methyl, methyl vinyl, methyl phenyl, and trifluoropropylmethylvinyl silicone rubber without silica fillers in the spiral-coiled membrane lung.

Spiral-coiled membrane lungs of 0.6 m2 surface area were fabricated from silicone rubber membranes coated with four types of commercially available pure-gum polysiloxane rubber: 1) polytrifluoropropylmethylvinylsiloxane, 2) polymethylphenylsiloxane, 3) polymethylvinylsiloxane, and 4) polydimethylsiloxane. Membranes lungs were tested in an arteriovenous shunt without added heparin after bypass was begun. All group 4 had major rise in resistance to blood flow or clotting in over 33% of the membrane lungs studied, after 6 hr of bypass. Membrane lungs with polydimethylsiloxane (group 4) had no rise in resistance to blood flow during 24 hr of bypass and had the least amount of changes in blood platelet count.


Progress in splat-cooling of dental alloys.

Improved techniques in preparing and sizing dental alloy powders containing Ag, Sn, and Cu have led to amalgams with higher strengths and less spread in dimensional change but very poor shelf life. Since maximum strengths occurred for amalgams containing about 15% more Sn than previously reported, and consequently presumably more of the weak gamma2 phase, the higher strengths are difficult to understand. The poor shelf life may be explained by oxidation of the highly reactive splat-cooled alloy or possibly by the existence of highly reactive metastable phases which transform fairly rapidly to the less reactive equilibrium ones.


Influence of cold plastic deformation on critical pitting potential of AISI 316 L and 304 L steels in an artificial physiological solution simulating the aggressiveness of the human body.

The effect of cold working on critical pitting potential of AISI 316 L and 304 L steels in a buffered physiological solution has been studied. In particular, the importance of deformation degree, orientation of the specimen surface to the deformation direction, and cold working temperature in lowering the critical pitting potential is shown.


Wear of dental amalgam.

The wear of cental amalgam was studied by single- and double-pass sliding and by a silicon carbide abrasion test. A dispersed amalgam showed significantly better resistance to two-body abrasion than the spherical amalgams tested. The wear amalgam was determined by resistance to penetration and by a ductile mode of surface failure over the load range studied. Differences in the wear of amalgam and restorative resins and composites are discussed.


Analysis of biomaterials deposited on soft contact lenses.

Formation of opaque deposits on the anterior (air) surface of hydrophilic soft contact lenses is a problem worthy of investigation by all concerned. These deposits have been analyzed for biomaterials by chemical, biochemical, electrophoretic, and immunological techniques. Qualitative and quantitative chemical colorimetric tests revealed the presence of variable amounts of protein (5-10 microgram/lens), carbohydrate (1.0-1.2 microgram/lens), and phospholipids (0.01-0.05 micronmole/lens). Cholesterol and glucose were not present at detectable levels. Fluorescent antibody tests with appropriate controls gave positive tests for albumin, lysozyme, gamma-G-globulin, and alpha1-lipoprotein in the deposits, all proteins present in tear fluid. Deposits were most effectively  removed from the lenses by the combination of heat, sodium dodecyl sulfate (SDS) detergent, and the thiol reagent dithiothreitol (DTT). SDS-denatured protein migrated on polyacrylamide gels with electrophoretic patterns corresponding to molecular weights for those proteins detected by the above antibody tests. The nature of the bonding interactions of biomaterials to the lenses was probed by chemical reagents used to remove them, employed singly and in all possible combinations. Urea, guanidine hydrochloride, potassium thiocyanate, potassium perchlorate, hydroxylamine, and EDTA were much less effective than SDS and DTT. These data suggest that apolar interactions plus disulfide bonds may be important in stabilizing the deposit structure, and point to improved cleaning procedures.


Crevice corrosion in orthopedic implant metals.

Studies of orthopedic implant alloys in crevice configuration suggest that cobalt-chromium alloys such as Haynes-Stellite 25 are the best choice for multicomponent implants. It is also shown that severe crevice attack may result from use of mixed metals.


Cellulosic ion-exchange membranes for hemodialysis.

The application of cellulosic ion-exchange membranes to hemodialysis was studied in vitro. The membranes were prepared by radiation-grafting methacrylic acid and vinylpyridine to films of DuPont cellophane PD-215 to produce cation-exchange and anion-exchange membranes, respectively. Solutions of urea, creatinine, glucose, and uric acid were studied for their interactions with and diffusion through the membranes. Ultrafiltration rates were also determined. Cuprophane and PD-215 cellophane were studied as controls. Dialysis plots for the membranes revealed a mechanism of "assisted transport." Initially, the solutes were removed from solution by a sorption/adsorption mechanism followed by a steady-state diffusion process. The calculated diffusivities for these later steady-state regions increased linearly with capacity for urea, creatinine, and uric acid, while for glucose the reverse was true. The combined processes involved provided considerably greater mass transport per unit thickness than either DuPont PD-215 cellophane or Cupropane.


Analysis of the ingredients and determination of the residual components of acrylic bone cements.

Rapid and reliable methods for the characterization and quantitative determination of ingredients usually present in self-curing methacrylate bone cements were developed using spectrophotometric, gas chromatographic (GC), and coventional gravimetric procedures. These procedures are applicable to non-crosslinked methacrylate resins. In the presence of some copolymers, polymer blends or crosslinking agents, or other ingredients which will result in the formation of insoluble methacrylate resins, some modification of the identification procedures and quantitative estimation of the resin components will be required. Molecular weight and molecular weight distribution of the powder and cured cement were obtained from viscosity and gel-permeation measurements. Residual low molecular weight materials in the cured cement were determined for various storage times. Residual and water-leachable monomer and residual dimethyl-p-toluidine (DMPT) were measured by GC. In air, The monomer content of the cured resin decreased from 3.3% after 1 hr to 2.4% after 215 days. When stored in water at 37 degrees C, over 65% of the extractable monomer is leached out within 10 min. After 6 hr, the rate of diffusion of monomer into the aqueous phase is low. The cured cement contains as much as 0.6% DMPT, which does not leach out. Residual peroxide, estimated spectrophotometrically, amounts to 0.8%, but decreases to 0.14% after 15 months in water at 37 degrees C. A semiquantitative colorimetric test indicated that the hydroquinone content of specimens analyzed within 24 hr after curing amounted to 9 ppm.


Fracture of the hook of the hamate in athletes.

During an eight-year period, four tennis players, seven golfers, and nine baseball players were seen with a fracture of the hook of the hamate. Eighteen of these twenty patients were disabled by pain and after the fracture fragment was removed, all eighteen were relieved so that they returned to their athletic pursuits. Two patients were asymptomatic, their old fracture being discovered accidentally when they were treated for other injuries. Nineteen of the twenty patients had been examined before coming under our care, but the correct diagnosis had been made in only two. Conservative treatment, including rest, physical therapy, and injections of steroids into the wrist and hand, had not been beneficial. From the history and findings, we believe that these fractures were caused by a direct blow against the hook of the hamate caused by the handle of the tennis racket, golf club, or bat during a swing, and not by indirect force produced by the ligaments and muscles attached to the hook. The fracture was demonstrated in all twenty patients by a roentgenogram (profile view) of the carpal tunnel.


Ununited fracture of the hook of the hamate.

Of nine ununited fractures of the hook of the hamate, eight were treated by surgical excision of the fragment. All eight patients had relief from persistent pain and regained the preinjury level of function. This fracture occurs frequently in athletes and is often not diagnosed because it can be demonstrated only by special roentgenographic views. It is concluded that this fracture may be less rare than is commonly believed and that surgical excision permits early return of function, especially in athletes and laborers who must be able to grip hard objects strongly.


Supracondylar fractures of the humerus: the role of dynamic factors in prevention of deformity.

In a retrospective survey of eighty-two children with supracondylar fractures of the humerus, forty were found to have sufficient clinical and roentgenographic data to classify the fractures as varus or valgus, and to determine the carrying angles at end result. The initial displacement (varus or valgus) of the distal fragment correlated with the final carrying angle, but the use of internal fixation, the adequacy of reduction achieved and the duration of traction or immobilization did not. The sixteen varus supracondylar fractures immobilized with the forearm in supination had a mean varus deformity of about 16 degrees, whereas the eight immobilized in pronation had a varus deformity of only 3 degrees. The findings in the few valgus fractures suggested that the valgus angulation was less at follow-up if they had been immobilized in supination. Dissection of eight cadaver limbs and electromyographic studies of one normal limb suggested that the position of the forearm affects the result by altering muscle tension.


Functional bracing of fractures of the shaft of the humerus.

We treated fifty-one cases of fracture of the shaft of the humerus with a functional method of treatment consisting of a plastic sleeve, individually molded or prefabricated. It maintained good alignment of the fragments and permitted rapid and uninterrupted osteogenesis. The early introduction of functional activity to the entire extremity appears to provide a desirable physiological environment conducive to rapid healing. Non-unions have not been encountered in nonpathological fractures. Healing time has been rapid, and there has been consistent restoration of motion of all joints prior to the completion of healing. The morbidity was minimum.


The spherocentric knee: biomechanical testing and clinical trial.

The spherocentric knee is a non-hinged, intrinsically stable knee-joint prosthesis which allows controlled triaxial rotation and was designed specifically for use in severely deformed or grossly unstable knees. Biomechanical evaluation of the prosthesis implanted in fresh human cadaver limbs demonstrated highly favorable motion, stability, strength, deflection, and energy absorption characterists for the spherocentric prosthesis-bone assembly as compared with control cadaver knees. After an average follow-up of 25.5 months after the first twenty-five spherocentric arthroplasties, the clinical results were highly satisfactory and suggested that the spherocentric knee may be relatively resistant to the problems of wear debris, loosening, and breakage which have plagued other intrinsically stable knee-joint prostheses.


Painful knees in older patients: radionuclide diagnosis of possible osteonecrosis with spontaneous resolution.

The syndrome of osteonecrosis of the femur at the knee presents as sudden onset of knee pain in elderly patients. It is classically associated with a subchondral radiolucency and a positive radionuclide bone scan. The lesion may progress to collapse of the bone. In this report, twelve cases are described in which the clinical entity of osteonecrosis of the femoral condyle was diagnosed because the patients had a clinical picture identical to that of osteonecrosis and also had positive bone scans. However, roentgenographic changes supposedly diagnostic of the condition were not seen. In twelve patients, arthrotomy was avoided. They became asymptomatic and their bone scans returned to normal in a few months.


Metal carcinogenesis: a study of the carcinogenic activity of solid metal alloys in rats.

The carcinogenic activity of seven metallic alloys was studied by implanting solid rods of each alloy in the gluteal muscles of Sprague-Dawley rats which were killed after twenty-four months. When the number of tumors in these rats was compared with the numbers in rats not operated on, in rats which had a sham operation, and in rats implanted with Silastic rods, no statistically significant differences in the incidences of the tumors in the several groups were found.


Damage to the capital femoral epiphysis due to Frejka pillow treatment.

Seven cases of damage to the capital femoral epiphysis in seven infants with congenital dysplasis, subluxation, or dislocation of the hip treated with a Frejka pillow splint are described. The pillow was first applied shortly after birth (up to four months of age) and was continued for three to six months. The roentgenographic changes in the femoral head and neck were consistent with either avascular necrosis of the femoral head or a growth disturbance causing alteration of the normal angle of the neck and shortening of the affected femur.


Determination of the vascularity of the femoral head with technetium 99m-sulphur-colloid.

The results of technetium 99m-sulphur-colloid scans performed at the Los Angeles County-University of Southern California Medical Center and Rancho Los Amigos Hospital to assess the vascular status of the femoral head after sixty-five displaced and thirty undisplaced fractures of the neck of the femur and forty-four traumatic hip dislocations as well as in fifty patients with idiopathic ischemic necrosis of seventy femoral heads were evaluated by histological and clinical examination after follow-up of two years or more. The accuracy of the scan as corroborated by the histological findings and clinical results was 95 per cent. The method is safe, reliable, accurate, and simple in contrast to other methods which have disadvantages related primarily to technical complexities, difficulty in interpretation, and expense.


Bone-mineral content of the femoral neck and spine versus the radius or ulna.

The interrelationship between the bone-mineral content of sites on the radius or ulna and that of the femoral neck and lower thoracic vertebrae of twenty-four human skeletons was determined. The average correlation coefficients, -/r, were 0.82 and 0.57. Regression analysis showed that the bone-mineral content of the radius provides a good estimate of the bone-mineral content of the femoral neck (standard error of estimate approximately 16 per cent). Multiple regression analysis using the age of the subject and the bone-mineral content of the radius provides an equation with a similar standard error of estimate for the vertebrae.


Effect of beta-aminopropionitrile on bone mechanical properties.

In vivo beta-aminopropionitrile treatment caused significant changes in the mechanical properties of rat femora. In femora treated with beta-aminopropinitrile as compared with controls, there was a 44 per cent decrease in bone stiffness and a 47 per cent increase in bone deformation to failure. Bone ash was decreased 5 per cent in the treated group. It is likely that the decrease in bone ash was secondary to impaired intermolecular bone-collagen cross-linking, which is a well established consequence of treatment with beta-aminopropionitrile. Thus, beta-aminopropionitrile influences bone mechanical properties directly by impairing collagen cross-linking and indirectly by altering bone-matrix mineralization.


Orthopaedic aspects of spinal tumors in children.

A retrospective survey has been made of forty children with spinal tumours. Difficulties in establishing the correct diagnosis are mentioned and the value of radiological and cerebrospinal fluid investigations discussed. The major orthopaedic disabilities are spinal deformity or instability, and paraplegia. The main factor in the development of the former is the site of laminectomy: the higher the level the greater is the likelihood of deformity or instability developing. Measures to prevent this distressing complication are discussed. The role of the orthopaedic surgeon in the management of these children is emphasised.


Sub-capital coxa valga after varus osteotomy for congenital dislocation of the hip.

Coxa valga may sometimes occur as a complication of varus osteotomy for congenital dislocation of the hip. Six such cases are described with a minimum follow-up of nine years. In three cases the varus osteotomy had been performed on only one side; in one case it was done on both sides and coxa valga developed bilaterally; and in two further bilateral cases coxa valga developed on one side only. In each case the coxa valga was sub-capital. Detailed radiographic analysis included measurements of neck-shaft angle, acetabular angle and C.E. angle. The epiphysis-shaft angle is described; it is an index of the constant tendency of the capital epiphysis to assume a horizontal position. The cause remains unknown, but damage to the trochanteric growth plate or to the lateral part of the capital growth plate could not be identified as aetiological factors. Poor acetabular cover was considered a possible factor. All the patients in this series had functionally excellent hips, but the long-term prognosis of the hips with partly uncovered femoral heads is doubtful.


A technique of correction and internal fixation for scoliosis.

This report describes a technique for the correction and fusion of scoliosis with the aid of a flexible metal rod that is fixed by wires under tension to the bases of the spinous processes on the convex side of the curve at a number of points. The results in a series of 100 cases are reported. In the last seventy patients, the average correction at one month was 51 per cent, and after two years 45 per cent. In these seventy cases the incidence of pseudarthrosis was 5-7 per cent.


The painful arc syndrome. Clinical classification as a guide to management.

Ninety-seven patients suffering from painful arc syndrome of the shoulder were studied. Local anaesthetic and radiographic contrast investigations were carried out. One-third of the patients had lesions in the posterior part of the rotator cuff which resolved after injections of local anaesthetic and steroid. One-third had anterior lesions in the subscapularis tendon: almost all resolved under the same regime but two required division of the coraco-acromial ligament. The remaining third had lesions of the supraspinatus tendon, usually associated with degeneration of the acromio-clavicular joint: most of these failed to gain relief from the local anaesthetic and steroid. Twenty-two operations were performed either by a transcromial or by a deltoid splitting approach. Excision of the outer end of the clavicle and division of the coraco-acromial ligament abolished the pain in most cases.


The morphometry and pathological anatomy of the lumbar spine in South African negroes and caucasoids with specific reference to spinal stenosis.

Direct measurements were made on 2,166 lumbar vertebrae of 433 adult negro and caucasoid skeletons. On statistical analysis, forty-five vertebrae in twenty-seven skeletons were found to be stenotic, the mid-sagittal diameter being the significantly reduced dimension. Whereas spinal stenosis syndromes are rare in South African negroes, the lumbar canal is marginally narrower in the negro. There is a uniformity of configuration and capacity of the lumbar spinal canal, which transcends race and sex. By a new method of determining the dorsal limit of the lumbar canal on lateral plain radiography, the overall average lower limit of normal of the mid-sagittal diameter is established at 15 millimetres, and of the transverse diameter 20 millimetres. Bony degenerative changes are more likely to cause neurological compression in the nerve root tunnel than in the spinal canal. The role of skeletal narrowing of the spinal canal as an exclusive cause of the spinal stenosis syndrome may have been exaggerated.


Diffusion of antibiotics from acrylic bone-cement in vitro.

The diffusion of Fucidin, gentamicin, and clindamycin from acrylic cement was tested in an in vitro system. The activity of Fucidin was very short-lived and only against gram-positive organisms; gentamicin inhibited gram-positive and gram-negative organisms for twenty-two and eleven days respectively; clindamycin had significant action only against gram-positive organisms and retained some activity for fifty-six days. We suggest that the destruction of organisms in the tissues is more likely to be achieved by topical and intravenous administration of antibiotics during the operation than by incorporation of antibiotic in the cement.


Antibiotic-loaded acrylic cement.

Laboratory experiments and clinical investigations have confirmed the various claims made originally by Buchholz and Engelbrecht (1970) that antibiotic-loaded acrylic cement releases the antibiotic into the surroundings in useful concentrations. Palacos R cement released higher concentrations than CMW, Simplex and Sulfix brands of cement and over longer periods. Concentrations of gentamycin and fucidin were sufficient to penetrate dead cortical bone. These conclusions need to be assessed with animal studies, mechanical testing and clinical results before the ideal place of antibiotic-loaded acrylic cement is established.


The ultrastructure of normal digital flexor tendon sheath and of the tissue formed around silicone and polyethylene implants in man.

Three normal digital flexor tendon sheaths and the corresponding tissue formed around five silicone rod tendon implants, two silicone rubber mammary prostheses and one polyethylene tubing implant have been examined by light microscopy and by transmission and scanning electron microscopy. No principal difference in morphology was found. The surface facing tendon or implant was almost invariably covered with an irregular layer of amorphous material and filaments; only occasionally were collagen fibrils or cells exposed. Beneath the surface there were abundant collagen fibrils and some cells; besides fibroblasts, cells rich in filaments and often with numerous glycogen granules, mitochondria and peripherally located vesicles were found. These cells were frequently surrounded by a thick layer of an amorphous matrix. The results indicate that the implants caused remarkably little tissue reaction.


Chondrosarcoma of the bones of the hand.

Nineteen chondrosarcomas are reported arising in proximal phalanges or metacarpal bones of the hand mainly in elderly patients, predominantly women. The usual clinical presentation was of a progressively painful large tumour, often arising in a dormant lesion near the metacarpo-phalangeal joint. Radiologically most showed some bone expansion with a poorly defined area of destruction and a considerable soft-tissue swelling. Histologically, malignancy was usually obvious, but confusion might arise from the inclusion of bland areas of chondromatous tissue that probably represented the original lesion. Four tumours, initially curetted and grafted, recurred locally and necessitated amputation of the digit or ray. Amputation was the primary treatment for fourteen other tumours and was curative except in one patient who eventually needed amputation through the forearm for a large second recurrence. One tumour was satisfactorily controlled by excision of the affected phalanx. None of these nineteen tumours is known to have metastasised. Correct treatment implies a carefully considered balance between conservation of function and complete removal of all tumour tissue.


Conformity in condylar replacement knee prosthesis.

Experiments were carried out to determine the optimum conformity between the femoral and tibial condyles in condylar replacement knee prostheses. Wear tests and observations from removed prostheses indicated that both high and low conformity produced characteristic abrasion and fatigue. Partly conforming condyles provided stability under load-bearing but allowed laxity to occur. Fixation to resist the various forces on the tibial components was enhanced by a short central intramedullary peg. Partial conformity is proposed as the optimum configuration between femoral and tibial components.


Polytenosynovitis caused by Toxoplasma gondii.

Two cases are reported of polytenosynovitis involving the wrists and ankles and caused by toxoplasmosis, together with the investigations that led to the diagnosis. Medical and surgical treatment of the first patient and medical treatment only of the second patient gave satisfactory results.


Posterior displacement osteotomy of the calcaneus.

In cases of established calcaneus after anterior poliomyelitis the deformity can be greatly reduced by combining an extensive plantar release with an oblique transverse osteotomy of the calcaneus that permits displacement upwards and backwards of the posterior weight-bearing part of the bone. The procedure greatly improves the mechanical advantage of subsequent tendon transplantations to the heel. Between 1956 and 1969 fifteen such osteotomies were carried out and the long-term results have been reviewed.


A new method of measuring patellar height.

To measure the patellar height the ratio of the articular length of the patella to the height of the lower pole of the articular cartilage above the tibial plateau is measured on a lateral radiography of the knee, flexed beyond 30 degrees. Normal values lie between 0-54 and 1-06. The subluxing patella is at the upper end of the normal range, but, in chondromalacia, the male patellae were lower than average, but the female patellae were normal.


Glycoprotein transport in the surface mucous cells of the rat stomach.

The intracellular transport of glycoproteins pulse-labeled in vitro with tritiated leucine and galactose in the surface mucous lining cells (SMC) of the fundus of the rat stomach was studied by electron microscope autoradiography. The SMC survive for several hours in pieces of the fundus incubated in a bicarbonate-buffered medium. The SMC have a normal ultrastructure for at least 4 h of incubation. Kinetic activity is normal for at least 5 h, as demonstrated by the normal nuclear incorporation of tritiated thymidine; The SMC incorporate labeled leucine and galactose at normal rates up to 4 h and 6 h, respectively. In contrast to the SMC, the cells of the gastric glands show signs of degeneration within 1 h after the start of incubation. In the SMC the secretory protein forms a smaller part of the total protein synthesized than in other secretory cells studied. The intracellular tranpsort of the leucine-labeled moiety of the glycoproteins follows the normal pathway. The RER loses 35% of its transportable labeled protein within 30 min. The Golgi complex is maximally labeled at 40 min and the mucous granules after 120 min. Galactose is attached to the glycoproteins mainly in the Golgi complex. Glycoproteins are not secreted within 2 h after synthesis of their protein moiety.


Release of spectrin-free vesicles from human erythrocytes during ATP depletion. I. Characterization of spectrin-free vesicles.

Human erythrocytes incubated without glucose at 37 degrees C (in vitro aging) release spectrin-free vesicles after 12 or more hours. The release of vesicles is dependent upon ATP depletion. If the endogenous level of ATP is maintained, vesicle release is completely inhibited up to 54 h. Vesicle release is independent of hemolysis because in vitro aged cells and cells that maintain their ATP levels lose identical amounts of hemoglobin up to 45 h. 93 percent of all membrane particles released constitute a uniform population of spheres with a diameter of 185 +/- 23nm. These vesicles are of slightly varying densities due to varying contents of hemoglobin. Vesicles contain half the amount of membrane protein that is found in intact membranes when referred to the content of phospholipids phosphorus. This is primarily due to the absence of spectrin. However, their content of protein component III, glycophorin, and cholesterol remains the same as in intact membranes. Thus, the major integral membrane proteins are present in vesicles in similar quantities were surface area as in cells except for the enzyme acetylcholinesterase that is enriched up to twofold. The phospholipids composition of these vesicles is representative of the intact membrane except that the amount of phosphatidic acid is 10-fold higher and the amount of phosphatidylethanolamine is slightly lower than in erythrocytes. These results suggest a selective release of membrane domains that lack peripheral membrane proteins and are enriched in acetylcholinesterase. This release of spectrin-free vesicles from cells aged in vitro could represent an acceleration of the physiological aging process.


Mitogenic activity of pituitary hormones on cell cultures of normal and carcinogen-induced tumor epithelium from rat mammary glands.

Cell suspension containing normal or tumor epithelium were readily obtained by enzymatically digesting rat mammary glands from perphenazine-treated (prolactin-hypersecreting) cycling, female virgin animals or hormone- responsive mammary tumors from animal treated with dimethylbenzanthracene. Cell suspensions were fractioned into predominantly epithelial and predominantly stromal cells by their differential rates of attachment to culture dishes. Both normal mammary and tumor epithelial cells were characterized by the presence of specific cell-junctional complexes, desmosome-like structures, surface microvilli, and their ability to synthesize casein. Serum-dependent protease activity was greater in cultures derived from tumors, and cells from such cultures grew in agarose whereas those from the non-neoplastic gland did not. The addition of prolactin to the culture medium stimulated DNA synthesis in primary or secondary epithelial cultures from tumors, whereas additional insulin and hydrocortisone with prolactin were required for similar levels of DNA synthesis in cultures from non-neoplastic glands. The fraction of cells synthesizing DNA was, however, smaller than that with 10 percent serum measured in the same time period. Both growth hormone and epidermal growth factor stimulated DNA synthesis but to a lesser extent than did prolactin. Prolactin with hydrocortisone and insulin were relatively inactive in promoting DNA synthesis of the nonepithelial cells whereas pituitary fibroblast growth factor was more active. These mitogenic effects were obtained when the hormones were added to the medium at near physiological concentrations, and paralleled the known activities of the hormones in control of mammary gland growth and development in the rat.


Inhibitory effect of colchicine on amylase secretion by rat parotid glands. Possible localization in the Golgi area.

Colchicine inhibited amylase secretion by isolated rat parotid glands only 6 h after administration of the drug in vivo. This delayed effect was not the result of the inability of the drug to reach its reaction site. When parotid glands were emptied of their secretory granules by isoproterenol treatment, the subsequent replenishment of cells with granules was inhibited by colchicines. Colchicine concomitantly produced alterations of the Golgi complexes, the cisternae of which were reduced in size and surrounded by clusters of microvesicles. Incubation of parotid glands with colchicines for prolonged durations failed to alter stored amylase secretion as stimulated by isoproterenol, but it inhibited the release of de novo synthesized enzyme. Another colchicines-binding activity, firmly bound to the particular fraction of homogenates, was found, of which a part may represent membrane located microtubular protein. An assembly-disassembly cycle of microtubules appears to exist in the parotid gland, as in the liver. However, only 14 percent of tubulin was found to be polymerized as microtubules in parotid glands as opposed to 40 percent in the liver. The present data suggest that colchicine primarily inhibits the transfer of secretory material towards or away from the Golgi complexes but not the hormone-stimulated secretion of stored amylase.


Protein of insect sperm mitochondrial crystals. Crystallomitin.

Mitochondrial derivatives of insect sperm usually contain a crystalline protein that shows a 45-nm main period, made up of 20-nm subperiods, determined by the coiling of filament bundles. Filaments are 2 nm thick and have a globular appearance. The crystals contain two main polypeptides, 52,000 and 55,000 daltons. These polypeptides are closely related, contain a high percentage of proline, and are insoluble in sodium dodecyl sulfate due to disulfide cross links. We suggest for this class of protein the name crystallomitin.


Nuclear matrix. Isolation and characterization of a framework structure from rat liver nuclei.

A nuclear framework structure termed the nuclear matrix has been isolated and characterized. This matrix forms the major residual structure of isolated nuclei and consists largely of protein with smaller amounts of RNA, DNA, carbohydrate, and phospholipid. The nuclear matrix can be further resolved by combined treatment with DNase and RNase. The remaining nuclear protein structure, after extraction of 90 percent of the nuclear protein, 99.9 percent of the DNA, and 98 percent of the RNA and phospholipid, is termed the nuclear protein matrix. Electron microscopy of this final nuclear protein matrix reveals an interior framework structure composed of residual nucleolar structures associated with a granular and fibrous internal matrix structure. The internal matrix framework is derived from the interchromatinic structures of the nucleus, and is connected to a surrounding residual nuclear envelope layer containing residual nuclear pore complex structures. Sodium dodecyl sulfate-acrylamide gel electrophoresis of the nuclear matrix proteins demonstrates three major polypeptide fractions, P-1, P-2, and P-3, with average molecular weights of approximately 69,000, 66,000 and 62,000, as well as several minor polypeptides which migrate at approximately 50,000 and at higher molecular weights (>100,000). Polypeptides with molecular weights identical to those of P-1, P-2 and P-3 are also components of isolated nuclear envelopes and nucleoli, whereas isolated chromatin contains no detectable matrix polypeptides. This suggests that the major matrix polypeptides are localized in specific structural regions of the nucleus, i.e., nuclear envelope, nucleoli, and interchromatinic structures. The presence of cytochrome oxidase activity in the isolated nuclear matrix indicates that at least some integral proteins of the nuclear membrane are associated with the matrix.


On the mechanism of ATP-induced shape changes in human erythrocyte membranes. I. The role of the spectrin complex.

Human erythrocyte ghosts have been shown, by scanning electron microscopy, to undergo ATP-dependent shape changes. Under appropriate conditions the ghosts prepared from normal disk-shaped intact cells adopt a highly crenated shape, which in the presence of Mg-ATP at 37 degrees C is slowly converted to the disk shape and eventually to the cup shape. These changes are not observed with other nucleotides or with 5'-adenylyl imidodiphosphate. Anti-spectrin antibodies, incorporated along with the Mg-ATP into the ghosts in amounts less than equivalent to the spectrin, markedly accelerate the shape changes observed with the Mg-ATP alone. The Fab fragments of these antibodies, however, have no effect. The conclusion is that the structural effect produced by the ATP is promoted by the cross-linking of spectrin by its antibodies, and may therefore itself be some kind of polymerization or network formation involving the spectrin complex on the cytoplasmic face of the membrane. The factors that contribute to the shape of the ghost and of the intact erythrocyte are discussed in the light of these findings.


Terminal phase of cytokinesis in D-98s cells.

The events leading to the completion of cytokinesis after the formation of the midbody and intercellular bridge in D-98S cells were studied with light and electron microscopy. Pairs of daughter cells corresponding to different stages of cytokineses, as determined previously form time lapse films, were selected from embedded monolayers for serial sectioning. Separation of daughter cells is preceded by the reduction in diameter of the intercellular bridge from 1-1.5 mum to approx. 0.2 mum. Two processes contribute to this reduction: (a) The intercellular bridge becomes gradually thinner after telophase; a progressive breakdown of midbody structures accompanies this change; and (b) the more significant contribution to reduction in bridge diameter occurs through the localized constriction of a segment of the intercellular bridge.. The microtubules within the constricted portion of the bridge are forced closer together, and some microtubules disappear as this narrowing progresses. The plasma membrane over the narrowed segments is thrown into a series of wavelike ripples. Separation of daughter cells is achieved through movements of the cells which stretch and break the diameter-reduced bridge. The midbody is discarded after separation and begins to deteriorate. Occasional pairs of daughter cells were found in which incomplete karyokineses resulted in their nuclei being connected by a strand of nuclear material traversing the bridge and midbody. Such cells do not complete cytokinesis but merge together several hours after telophase. This merging of daughter cells coincides with the nearly complete breakdown of the midbody.


Movements and associations of ribosomal subunits in a secretory cell during growth inhibition by starvation.

In Chironomus tentans salivary gland cells, the cytoplasm can be dissected into concentric zones situated at increasing distances from the nuclear envelope. After RNA labeling, the newly made ribosomal subunits are found in the cytoplasm mainly in the neighborhood of the nucleus with a gradient of increasing abundance towards the periphery of the cell. The gradient for the small subunit lasts for a few hours and disappears entirely after treatment with puromycin. The large subunit also forms a gradient but one which is only partially abolished by puromycin. The residual gradient which which is resistant to the addition of the drug is probably due to the binding of some large ribosomal units to the membranes of the endoplasmic reticulum (J.-E. Edstrom and u. Lonn. 1976. J. Cell Biol. 70:562-572, and U. Lonn and J.-E. Edstrom. 1976. J. Cell. Biol. 70:573-580). If growth is inhibited by starvation, only the puromycin-sensitive type gradient is observed for the large subunit, suggesting that the attachment of these newly made subunits to the endoplasmic reticulum membranes will not occur. If, on the other hand, the drug-resistant gradient is allowed to form in feeding animals, it is conserved during a subsequent starvation for longer periods than in control feeding animals. This observation provides a further support for an effect of starvation on the normal turnover of the large subunits associated with the endoplasmic reticulum. These results also indicate a considerable structural stability in the cytoplasm of these cells worth little or no gross redistribution of cytoplasmic structures over a period of at least 6 days.


Relationships between levels of membrane-bound glucuronidase and the associated protein egasyn in mouse tissues.

Mouse beta-glucuronidase has a dual intracellular localization, being present in both endoplasmic reticulum and lysosomes of several tissues. Previous studies demonstrated that the protein egasyn is complexed with microsomal but not lysosomal glucuronidase and that a mutant lacking egasyn is deficient in microsomal, but not lysosomal, glucuronidase. By means of a recently developed radioimmunoassay for egasyn, the relationship between microsomal glucuronidase levels and egasyn levels has been examined in various adult tissues, during postnatal development in liver, and after androgen induction of glucuronidase in kidney. The results indicate that the relative availability of egasyn determines the balance between glucuronidase incorporation into membranes and that into lysosomes.


Annulate lamellae in phloem cells of virus-infected Sonchus plants.

The occurrence of annulate lamellae (AL) in differentiating phloem of Sonchus oleraceus (Compositae) singly infected with sowthistle yellow vein virus (SYVV) and doubly infected with a combination of SYVV and beet yellow stunt virus is documented by electron microscopy. Cell types in which AL were found were immature sieve elements and phloem parenchyma cells. AL were found only in cells that also contained SYVV particles although a direct association between the virus and AL was not apparent. The substructure of the AL and the relationships between the AL and the nuclear envelope and endoplasmic reticulum  are similar to those reported in other descriptions of this organelle in the literature. This report appears to be the first one concerning the association of AL with a plant virus disease.


Changes in cell surface and cortical cytoplasmic organization during early embryogenesis in the preimplantation mouse embryo.

Membrane topography and organization of cortical cytoskeletal elements and organelles during early embryogenesis of the mouse have been studied by transmission and scanning electron microscopy with improved cellular preservation. At the four- and early eight-cell stages, blastomeres are round, and scanning electron microscopy shows a uniform distribution of microvilli over the cell surface. At the onset of morphogenesis, a reorganization of the blastomere surface is observed in which microvilli becomes restricted to an apical region and the basal zone of intercellular contact. As the blastomeres spread on each other during compaction, many microvilli remain in the basal region of imminent cell-cell contacts, but few are present where the cells have completed spreading on each other. Microvilli on the surface of these embryos contain linear arrays of microfilaments with lateral cross bridges. Microtubules and mitochondria become localized beneath the apposed cell membranes during compaction. Arrays of cortical microtubules are aligned parallel to regions of apposed membranes. During cytokinesis, microtubules become redistributed in the region of the mitotic spindle, and fewer microvilli are present on most of the cell surface. The cell surface and cortical changes initiated during compaction are the first manifestations of cell polarity in embryogenesis. These and previous findings are interpreted as evidence that cell surface changes associated with trophoblast development appear as early as the eight-cell stage. Our observations suggest that morphogenesis involves the activation of a developmental program which coordinately controls cortical cytoplasmic and cell surface organization.


Choline acetyltransferase activity of spinal cord cell cultures increased by co-culture with muscle and by muscle-conditioned medium.

Activity of the enzyme choline acetyltransferase (CAT), which mediates the synthesis of the neurotransmitter, acetylcholine, was increased up to 20- fold in spinal cord (SC) cells grown in culture with muscle cells for 2 wk. This increase was directly related to the duration of co-culture as well as to the cell density of both the SC and muscle involved and was not affected by the presence of the acetylcholine receptor blocking agent, alpha-bungarotoxin. Glutamic acid decarboxylase (GAD) activity was often markedly decreased in SC-muscle cultures while the activities of acetylcholinesterase and several other enzymes were little changed. Increased CAT activity was also observed when SC cultures were maintained in medium which had been conditioned by muscle cells or by undifferentiated cells from embryonic muscle. Muscle-conditioned medium (CM) did not affect the activities of SC cell GAD or acetylcholinesterase. Dilution or concentration of the CM directly affected its ability to increase SC CAT activity , as did the duration and timing of exposure of the SC cells to the CM. The medium could be conditioned by muscle cells in the presence or absence of serum, and remained effective after dialysis or heating to 58 degrees C. Membrane filtration data were consistent with the conclusion that the active material(s) in CM had a molecular weight in excess of 50,000 daltons. We conclude that large molecular weight material that is released by muscle cells is capable of producing a specific increase in CAT activity of SC cells.


Quantitation of nexus junctions in the granulosa cell layer of rabbit ovarian follicles during ovulation.

Electron microscopy was used in a semi-quantitative study to determine changes in the abundance and size of surface nexuses and changes in the abundance of interiorized nexuses in growing and mature ovarian follicles during the ovulatory process. Mature follicles contain larger granulosa cells than follicles in the early stage of antral formation. Also, the granulosa cells of mature follicles have a slightly greater number of surface nexuses (without a change in nexus length), and more interiorized nexuses, compared to immature follicles. As a mature follicle approaches rupture, there is an appreciable decrease in the number of surface nexuses per granulosa cell. There is also a slight reduction in the number of interiorized nexuses at this time. It is concluded that this decrease in both surface nexuses and interiorized nexuses may be a  consequence of ovulatory changes during which the rate of granulosa cell division is greater than the rate of formation of new nexuses. Additionally, the disruption to cell-to-cell cohesion during the ovulatory process appears to be independent of the interiorization of surface nexuses.


The effect of oxygen and vitamin E on the lifespan of human diploid cells in vitro.

Human diploid cells (WI-38) were serially subcultivated at partial pressures of oxygen (Po2) ranging from 5.6 mm Hg to 608 mm Hg. At a Po2 of 5.6 mm Hg, the number of doublings to phase out was less than that of control cells at a Po2 of 137 mm Hg. Cultures grown at Po2's of 24, 49, or 137 mm Hg grew at the same rate and phased out after a similar number of population doublings. Population lifespan was markedly shortened by chronic exposure to elevated Po2's, a phenomenon that was, in part, reversible. d-1-alpha-Tocopherol (10 microgram/ml or 100 microgram/ml) homogenized into the medium at each weekly subcultivation did not extend the lifespan of cells at reduced, ambient, or elevated oxygen tensions. These results indicate that neither oxygen toxicity nor free radical reactions play a significant role in limiting the lifespan of WI-38 cells grown in vitro under ambient oxygen tensions (Po2 137 mm Hg).


Spermatogenic cells of the prepuberal mouse. Isolation and morphological characterization.

A procedure is described which permits the isolation from the prepuberal mouse testis of highly purified populations of primitive type A spermatogonia, type A spermatogonia, type B spermatogonia, preleptotene primary spermatocytes, leptotene and zygotene primary spermatocytes, pachytene primary spermatocytes and Sertoli cells. The successful isolation of these prepuberal cell types was accomplished by: (a) defining distinctive morphological characteristics of the cells, (b) determining the temporal appearance of spermatogenic cells during prepuberal development, (c) isolating purified seminiferous cords, after dissociation of the testis with collagenase, (d) separating the trypsin-dispersed seminiferous cells by sedimentation velocity at unit gravity, and (e) assessing the identity and purity of the isolated cell types by microscopy. The seminiferous epithelium from day 6 animals contains only primitive type A spermatogonia and Sertoli cells. Type A and type B spermatogonia are present by day 8. At day 10, meiotic prophase is initiated, with the germ cells reaching the early and late pachytene stages by 14 and 18, respectively. Secondary spermatocytes and haploid spermatids appear throughout this developmental period. The purity and optimum day for the recovery of specific cell types are as follows: day 6, Sertoli cells (purity>99 percent) and primitive type A spermatogonia (90 percent); day 8, type A spermatogonia (91 percent) and type B spermatogonia (76 percent); day 18, preleptotene spermatocytes (93 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent).


Organization of cell junctions in the peritoneal mesothelium.

Intercellular junctions in the mesothelium of the visceral (mesentery and omentum), and parietal (diaphragm, pre-aortic, and iliac region) peritoneum were examined in rats and mice by using freeze-cleaved preparations. In addition to usual intercellular junctions (cell body junctions), special junctions are found between cell processes and the surface of the neighboring cell (cell process junctions). Cell body junctions are provided with tight junctions and communicating (gap) junctions. The former consist of one to two junctional strands which show a characteristic staggered arrangement, and focal discontinuities. In cell process junctions, the strands form loops or appear as short, free-ending elements; their polymorphism suggests considerable lability, probably in connection with their assembly and disassembly. The existence of free-ending strands indicates that such structures can be used as attachment devices without being concomitantly involved in the formation of occluding zonules. In both types of junctions, the strands can be resolved into bars, approximately 80- 100nm long, frequently provided with terminal enlargements and intercalated particles which occur singly or in small clusters. These particles are morphologically similar to those present in communicating (gap) junctions. The mesothelium is also provided with isolate composite macular junctions. Throughout the mesothelium, the cleavage plane follows the outer contour of junctional strands and particles, suggesting that strand-to-strand interactions in the apposed membranes are weaker than interactions between each strand and underlying cytoplasmic structures. In their general geometry and cleavage characteristics, the mesothelial junctions resemble the junctions found in the venular endothelium.


[Generalised fecal peritonitis. Etiopathogenic and therapeutic considerations in 30 cases (author's transl)].

The authors analyse a series of 30 cases of generalised fecal peritonitis which represents a less frequent form of peritoneal infection. The etiology was mainly diverticular perforation (50 p. cent) or neoplasia (25 p. cent). The mortality was high (63 p. cent). It seems, in the light of the results, that the best treatment consists of operating in two stages, the first being the eradication of the colonic lesion which should be carried out as a routine without attempting immediate anastomosis. Wide drainage with peritoneal lavage and the use of "noxythioline" may improve the results of a disease which always remains very severe.


[Treatment of fractures of the cervical spine and of the dorso lumbar spine with neurological complications (author's transl)].

Our surgical therapeutic attitude in neurological complications of fractures of the spine seems to us justified by the results which we have obtained. It never caused any aggravation and permitted better relief of the complications of bed rest during the early stages. It always permitted satisfactory vertebral position. It is of course difficult to say definitely whether it was responsible for the neurological recovery often observed. It is nevertheless probable in a certain number of cases and in any case, this recovery will occur under better conditions more favourable for later rehabilitation. But one should emphasise if one wishes to improve further the results, that this reduction should be performed very rapidly.


[Peri-ano-rectal cysts and tumours of vestigial origin in the adult (author's transl)].

Four cases of cysts or tumours developing from embryological remnants in the peri-ano-rectal space are reported. The authors propose for these rare lesions a coherent and simplified classification in two main groups: teratomas on the one hand, developmental cysts on the other hand. The various methods of diagnosis are studied. The precise diagnosis of a congenital cyst can only be made by pathological examination of the specimen after surgical removable. Any rapid local change implied malignant degeneration. Such a risk suggests immediate removal of any known ano-rectal cyst or tumour. The route of approach is usually perineal. Kraske's position gives good access to the retro-ano-rectal space. Possible pelvic spread of these lesions sometimes requires the use of the abdominal approach, either straight away or secondarily when total removal is impossible by the perineal route alone.


[Acute postoperative arterial ischemia (author's transl)].

The authors analyse 16 cases of acute ischemia observed over a period of one month after an operation of general surgery. The classical cardiac origin, e.g. infarct, arrythmia, was found in only three patients. The septic etiology was predominant, septic arterial emboli but not infective or acute thrombosis during septic or post-operative shock. The clinical aspect showed no special characteristics. Surgery remains the principal form of treatment. The results are good, except in acute arterial thrombosis justifying plastic surgery which may give better results than simple thrombectomy.


[Wounds of the hand by injection of fluids under high pressure (author's transl)].

The result of mechanisation and technical improvements in industry, wounds by high pressure pistols are very serious. They are often neglected at first for they may be undiagnosed. To the toxicity of the injected product, which diffuses far from the minimal point of entry, may be added the destructive effect of the high pressure on the cell spaces of the hand, which are narrow and inextensible. Surgical treatment includes the association of antibiotics, anti-inflammatory drugs, analgesics and wide removal of the affected tissues as well as the foreign material. Active physiotherapy after immobilisation in a functional position, will permit recovery of satisfactory movements. Amputation is unfortunately often necessary, thus great efforts should be made to prevent work accidents.


[Primary cancer of the gallbladder. Analysis of a surgical series of 24 cases. Attempted anatomical classification of its prognosis. Proposal of a preventive approach].

This analysis of 24 cases of primary carcinoma of the gall bladder is compared with series in the literature. The extremely poor prognosis of this carcinoma, recognised by all authors, may be explained by the unfavourable background, delay in diagnosis at all stages, early regional spread, difficulty in laying down the therapeutic indications. A Pathological classification for prognosis has been proposed for the surgeon with stages: O, I, II, II a and b, IV a and b, in relation to the spread of the gall bladder carcinoma and reflexions inspired by analysis of the operative results in the literature. Even with better knowledge of predisposing factors, and if new methods of treatment gave greater hope in the future, more frequent use of cholecystectomy seems the best prevention of bladder carcinoma in any case of cholelithiasis whether symptomatic or not, in the absence of any contra-indication.


[Familial colonic polyposis (author's transl)].

During the last 5 years, 7 families suffering from familial polyposis coli have been investigated; we found 23 members affected and we operated 10 of them. One of them died of metastases and 9 members of these pedigrees died of carcinoma which appeared at the average age of 41,5 years. This confirms that this condition is the most serious precancerous illness and needs regular controls of all members of these families. At the occasion of this review, we describe the main features of this condition and briefly our surgical attitude.


[New acquisitions on the arterial vascularization of the 3d and 4th portions of the duodenum. Surgical applications].

The authors studied the blood supply of the 3rd and 4th parts of the duodenum in 15 adult subjects. Although the duodeno-pancreatic arches are constantly found, the branches to the duodenum have a variable lay-out. On the right, the superior mesenteric artery, the distribution of which is always regular on both sides of the 3rd part of the duodenum. On the other hand, on the left, the superior mesenteric artery, there is sometimes a vascular hiatus which may extend to the duodeno-jejunal angle. Although in 60 p. cent of cases this part of the duodenum is well supplied with blood on both surfaces, in 40 p. cent of cases on the other hand, there exists an avascular area on both sides. This lay-out is not favourable for division and anastomsis of this segment of the digestive tract.


[Late urological complications of reconstructive surgery of the aortic bifurcation (author's transl)].

Ureteral obstruction following aorto-iliac or aorto-femoral dacron prosthesis has rarely been described. It seems, however, as shown in our three cases, that this complication is more common than was previously believed. In all three cases, ureteral obstruction was attributed to retroperitoneal fibrosis enclosing the graft. The threat of such a complication should lead the surgeon to carry out intravenous urography in all patients with a vascular prosthesis.


[General anesthesia in rabbits for digestive surgical research. A spontaneous ventilation technic by Nembutal-diazepam combination].

General anesthesia in the rabbit in spontaneous ventilation, followed by normal awakening and prolonged survival, has been studied here in order to have a series of chronic preparations which the authors could use in their research on digestive surgery. These type of anesthesia requires less personnel, but raises certain difficulties due to the closeness of the thresholds of efficacy and toxicity, the length of the experiment and program requirements. The reliable technic exposed here permits prolonged anesthesia (up to eight hours) with normal survival of the animals. The mortality due to anesthesia was low.


Separation of estrogen glucorinides, sulphates and phosphates on ion-exchange cellulose by high-pressure liquid chromatography.

In continuation of previous work, the selectivity of anion-exchange cellulose systems with respect to thirty estrogen conjugates was studied. The influence of the type and particle size of cellulose anion exchangers, the type and concentration of the counter anion, pH and temperature was investigated. On the basis of the results the optimal choice of the ion-exchange material, the eluent and the operating conditions is demonstrated. The results of the optimization are exemplified by a number of chromatograms showing rapid separations, including the determination of the estrogen conjugate profile of human pregnancy urine.


Use of a modified Tenax GC column packing for the direct gas chromatographic analysis of phenols in water at the ppm level.

Dilute solutions of phenols in water at the ppm level may be analysed gas chromatographically by direct injection onto a column with a packing similar to that used in the Viking 1975 spacecraft: Tenax GC (2,6-diphenyl-p-phenylene oxide polymer) modified with polymetaphenyl ether liquid phase to eradicate irreversible adsorption. Symmetrical peaks were observed for injections of less than 1 ng of phenol and some of its alkylated derivatives. Chromatographic properties of modified Tenax GC columns are reported along with their application to the analysis of some industrial effluents with previous work-up.


Bonded peptide stationary phases for the separation of amino acids and peptides using liquid chromatography.

Bonded optically active tripeptides have been applied as stationary phases for liquid chromatography. Significant retention variations are shown for some UV absorbing amino acids when compared to a silica gel column using the same mobile phase. The separation of certain isomeric dipeptides has been accomplished using  the bonded optically active tripeptide stationary phases. It is shown that these separations are superior to those possible on silica gel, although the efficiencies of the column are low. Phenylthiohydatoin (PTH)-amino acids have also been analyzed using the bonded tripeptide L-Val-L-Ala-L-Ser on silica gel CT and 1% citric acid-water as the mobile phase. No two PTH-amino acids showed the same capacity ratio out of 25 tested and the separation of 15 in one isocratic run is presented. The implications and future for the use of bonded opitcally active peptides as stationary phases for liquid chromatography are discussed.


Single- and two-step extraction and thin-layer detection procedures for benzoylecgonine (cocaine metabolite) alone or in combination with a wide variety of commonly abused drugs in urine screening programs.

Three extraction procedures for the detection of benzoylecgonine (a major metabolite of cocaine) in urine are presented. Each technique has its advantages, depending on the needs of a clinical operation. Procedures I and II involve the use of ion-exchange resin-loaded paper. Procedure I has a sensitivity of 1 microng/ml and requires 20 ml of urine, and is recommended when the aim is to test for the abuse of cocaine only. Procedure II is a two-step extraction method in which a wide variety of abused drugs are extracted by the first step and the benzoylecgonine left in the aqueous buffer phase is extracted in the second step. The sensitivity for benzoylecgonine using this procedure is 2 microng/ml and it requires 20-50 ml of urine. Procedure III involves the direct extraction of benzoylecgonine using 5 ml of urine and has a sensitivity of 0.5 microng/ml.


Thin-layer chromatography of chlorinated catechols.

The thin-layer chromatography of catechol and six chlorinated catechols has been studied on five adsorbents. Acetic acid in the developing solvent was essential for the production of sharp zones. The best separation of the individual compounds occurred on alumina, whereas silica gel appeared to be good for group separation (clean-up) from other compounds.


Retention behaviour of a bonded reversed phase in a high-performance liquid chromatographic assay of serum theophylline.

A simple, rapid assay for serum theophylline is described. Optimisation of the high-performance liquid chromatographic system has allowed the analysis of minimun sample volume (50 micronl) in the shortest time (8 min) with good between-batch precision (C.V. = 8.3%)and accuracy (recovery ca. 100%). The behaviour of theophylline and other xanthine derivatives, theobromine, caffeine and 8-chlorotheophylline, on a bonded octadecyl reversed-phase system suggests that C18 phase effects solute separation by mixed retention mechanisms rather than by pure reversed-phase chromatography. This behaviour is attributed to the participation of underivatised silanol groups in the chromatographic process, and observation corroborated by independent investigation. The mechanism of the silanol participation is difficult to rationalise; however, a normal-phase  liquid partition system complies with the experimental data.


Comparison of non-biospecific effects in immunoaffinity chromatography using cyanogen bromide and bifunctional oxirane as immobilising agents.

Polypeptide antigen, glucagon, antibodies to glucagon and non-immune globulins were immobilised on agarose using CNBr and a bifunctional oxirane. Irrespective of the ligand immolilised, positively charged groups introduced to conjugates by CNBr caused electrostatic interactions with impurities and soluble biospecific ligands. Solvents required for elution of bound antibodies and antigens were more strongly deforming when immunoaffinity conjugates were prepared with CNBr than with the oxirane. This is attributed to compound affinity resulting from reinforcement of biospecific by non-biospecific interactions. Strongly deforming solvents were still required for oxirane conjugates, however, when antibodies had high affinity for antigen.


High-pressure chromatography of tobacco mosaic virus on Spheron gels.

High-pressure chromatography on Spheron 300 BTD and Spheron 1000 was used to separate tobacco mosaic virus from other components of an extract from infected tobacco leaves, including F1, a protein having a molecular weight of 0.58 X 10(6). Comparative experiments have shown that the virus is not adsorbed by the carrier and is eluted in the free volume of the column; the virus is free from contamination by vegetable-juice components and is regained quantitatively. The virus can be separated partially, but distinctly, from the F1 protein by using a column of microgranular Spheron 300. Polyhydric phenols in the vegetable extract are adsorbed on Spheron and eluted from the column after a delay.


Method for improved protein separation in human and animal sera.

A method of improved protein separation in human and animal sera on columns of diethylaminoethyl cellulose is described. The improvement in protein separation is achieved by employing suitably modified elution gradients, the construction of which is described. A comparison with other work shows that improved resolution has been achieved over shorter elution times.


Somatostatin modulation of pancreatic glucagon, insulin, glucose and free fatty acids following beta-adrenergic stimulation.

This study was undertaken to determine the effect of somatostatin on acute, orciprenaline mediated, beta-adrenergic stimulation of free fatty acids, blood glucose, insulin, and glucagon in healthy subjects. After orciprenaline and somatostatin insulin and glucagon decreased, whereas blood glucose and free fatty acids increased, probably in part as a result of the lesser inhibition of glucagon (50%) than of insulin (83%). From these observations it is tentatively concluded that the inhibitory effects of somatostatin on insulin and glucagon release in man are a consequence of beta-adrenergic receptor involvement. These effects are possibly mediated through increased destruction of cAMP, blocking of camp dependent secretion or impairment of calcium uptake.


Thyroid function tests during the early phase of subacute thyroiditis.

Six patients with subacute thyroditis were followed with serial measurements of T4, FT4, TSH and RAI uptake. Five of the six underwent TRH stimulation early in the course of their illness. All six patients had elevated or high normal values for T4 and FT4 at the time of their clinical presentation (mean = 13.8 microgram per 100 ml and 3.9 ng per 100 ml, respectively). RAI uptakes were 1% or less in all six. TRH testing revealed a suppressed TSH response (mean deltaTSH less than0.1 muU/ml) in all five patients tested, suggesting hyperthyroidism. After initial studies were performed, five patients were treated with L-triiodothyronine (L-T3) and one with aspirin. All patients improved over a two to four week period of time, no relapses being noted.


Catecholamine metabolism in thyroid disease. II. Norepinephrine secretion rate in hyperthyroidism and hypothyroidism.

We have measured the secretion rate of norepinephrine (NE) in 6 euthyroid subjects, 6 hyperthyroid and 6 hypothyroid patients, infused at a constant rate (0.1 microC/kg/min) for 1 h with tritiated norepinephrine (New England Nuclear Inc.). Plasma NE concentrations were measured by a modification of the fluorometric method of Anton and Sayre. A significant linear relationship was observed between plasma NE and age in normal subjects. Accordingly, values for plasma NE have been corrected for age. Plasma NE concentration was 18.3 +/- 4.2 ng/100 ml (mean+/-SEM) in normal subjects compared with 17.5 +/- 3.3 ng/100 ml in hyperthyroid patients. There was a significant elevation of plasma NE (30.3 +/- 2.9 ng/100 ml) in hypothyroid patients (P less than 0.05). A significant elevation was observed in plasma NE secretion rates in hypothyroidism 4.62 +/- 0.98 microgram/kg/day (P less than 0.02) when compared to the control group (1.46 +/- 0.35 microgram/kg/day). No significant difference was observed between the hyperthyroid group (1.74 +/- 0.49 microgram/kg/day) and the controls. These data indicate that the plasma NE secretion rate is normal in hyperthyroidism, and is significantly elevated in hypothyroidism thereby explaining the higher plasma NE concentrations seen in hypothyroidism.


Increased plasma 11-deoxycorticosterone during spironolactone medication.

The increase of plasma deoxycorticosterone (DOC) levels after administration of spironolactone is a real effect and not due to cross-interference of the drug or its metabolites with the DOC-assay. This is proved by in vitro and in vivo results. Of all the metabolites only canrenone interferes to some extent with DOC after liquid-liquid-extraction and paper chromatography. The antiserum, however, is so highly specific, that the final cross-interference of the total procedure amounts to less than 2% of the DOC plasma levels. Moreover, in patients with Addison's disease only a very small increase of DOC plasma levels can be observed after administration of spironolactone in contrast to normal subjects.


Plasma 17alpha-hydroxyprogesterone in spironolactone-treated hypertensives.

When measured by a specific radio-assay method which excludes drug interference, plasma 17alpha-hydroxyprogesterone levels were found to be similar in spironolactone treated hypertensives, in untreated hypertensives, and in normotensive volunteers. No in vivo evidence for a significant inhibitory effect of spironolactone on adrenal 11beta, 17alpha and 21 hydroxylases was uncovered.


Change in affinity of insulin receptors following oral glucose in normal adults.

125I-insulin binding to circulating monocytes has been studied in 4 normal volunteers in the basal state as well as at 2 and 5 hours after ingestion of 100 g of glucose. In each study five hours after glucose ingestion, the competition-inhibition curve was shifted to the left and was steeper than that in the basal study; the amount of insulin that caused a 50% decrease in specific binding of 125I-insulin in the basal study was 3-11 fold higher than at 5 hours after glucose. These changes in binding after glucose ingestion were largely due to major alterations of receptor affinity. We conclude that acute changes in receptor affinity occur normally as part of the physiologic regulation of target cell sensitivity to hormonal stimulation.


Conversion of cortisol to cortisone by the human uterus and its reversal in pregnancy.

Inter-conversion of cortisol (F) and cortisone (E) was investigated by incubating minced tissue with tritiated cortisol or cortisone and then separating the products by Sephadex LH-20 column chromatography. In non-pregnant subjects conversion of F to E predominated (43.4+/-3.4% vs 0.1+/-0.4% for E to F). In early pregnancy F leads to E decreased and E leads to F rose while at term E leads to F (46.3+/-9.1%) exceeded F leads to E (15.1+/-6.8%). These results were in accord with those obtained by assaying the endogenous concentrations. In non-pregnant subjects the F/E ratio (1.1+/-0.6) was lower than that found in serum (6.3+/-2.2) while at term the uterine F/E (9.0+/-1.8) was similar to that of serum (8.8+/-2.0). These changes resulted in an 8-fold increase in uterine F compared with a 3-fold increase in serum F, while uterine E fell to 1/2 and serum E doubled. Thus, during pregnancy there is a dramatic reversal of the reaction in the uterus in favour of the active hormone. It seems possible that the increase in cortisol thus brought about may play an anti-immune role in uterine wall, the single tissue apart from blood in direct contact with fetal tissue.


Cortisol in human breast cancer tissue.

Endogenous cortisol levels were measured by radioimmunoassay in 140 primary and 31 metastatic human breast cancer specimens. The adjacent normal breast was assayed in 15 cases. Mean values for normal breast were 17.3 ng/g tissue. Significantly higher levels were found in cancer tissue. Primary lesions contained 26 ng/g and metastases 32.5 ng/g cortisol. These results lend support to our previous data on increased cortisol binding activity in human breast cancer.


The augmentation effects of secretin on the insulin responses to known stimuli: specificity for glucose.

Secretin stimulates insulin release directly and augments insulin responses when administered prior to an insulinogenic stimulus. The magnitude and specificity of this augmentation effect of secretin pretreatment was evaluated in normal subjects to pulses of the following stimuli: 5 g glucose (n = 6), 1 g arginine (n = 8), 2 microgram isoproterenol (n = 6), 0.5 g tolbutamide (n = 5) and 0.5 mg glucagon (n = 6). Secretin was administered either as a 15 U pulse with a 3 U/min infusion for 25 min or as 4-150 U pulses. A period of 30 min elapsed after either the cessation of the infusion or the final 150 U pulse. In all studies, the acute insulin responses immediately following secretin were observed and had returned to baseline levels prior to the administration of any insulinogenic stimulus. Secretin only augmented the acute insulin responses to 5 g glucose pulses (pre-secretin glucose; 28 +/- 20 muU/ml, mean +/- SD; post-secretin glucose pulse: 45 +/- 37 muU/ml, P less than 0.5), an increase of 56 +/- 21% (P less than .005). A highly linear relationship was noted (r = .98, P less than .01) between the acute insulin response to glucose before and after secretin suggesting that the insulin response to the control glucose pulse is a major determinant of the glucose stimulated response after secretin. Secretin pretreatment did not augment the insulin responses to arginine, isoproterenol, tolbutamide and glucagon. The specificity of secretin for augmenting glucose stimulated insulin release suggets a possible role for secretin in carbohydrate metabolism.


Effects of angiotensin II blockade by saralasin in normal man.

To evaluate the role of angiotensin II in normal man, sar1-ala8-angiotensin II (saralasin), a specific competitive inhibitor or angiotensin II, was given by graded iv infusion (1-20 microgram/kg/min over 1 h) to 6 healthy young volunteers. Significant angiotensin II-dependence of blood pressure was found only after the combined stimulus of sodium restriction (10 mEq daily for 4 days) and ambulation. Systolic blood pressure fell during saralasin infusion from 106.2 +/- 2.0 to 95.8 +/- 2.9 mm (P less than .05). At the same time, plasma renin activity rose strikingly from 3.3 +/- 0.6 to 21.6 +/- 8.5 ng/ml/h, and then fell to 9.3 +/- 4.9 over the post-infusion hour despite continued ambulation. Peak plasma renin activity correlated well with the simultaneous fall in blood pressure (r = .91; P less than .01), suggesting a compensatory response to the vascular effects of angiotensin blockaded. On an ad lib. diet, saralasin infusion had no consistent effect on plasma renin activity serum potassium, or blood pressure, but in supine volunteers plasma aldosterone increased from 9.8 +/- 2.1 to 16.5 +/- 4.8 ng/dl, an average increase of 55 +/- 17% (P less than .05). Although mena plasma cortisol decresed, individual changes in cortisol did not correlate with changes in plasma aldosterone. These data suggest that saralasin has an agonist effect on basal aldosterone secretion.


Secretion of parathyroid hormone by abnormal human parathyroid glands in vitro.

The secretory response of abnormal parathyroid glands obtained surgically from eleven patients with primary and secondary hyperparathyroidsm was tested in vitro. Short term flask studies were used to measure release of parathyroid hormone (PTH) at high (3.0 mM) and low (0.5 mM) calcium. Of eight adenomas, all but one showed increased release of hormone when exposed to low calcium ("responsive to calcium"), the degree of stimulation at three hours ranging from 15 to 209%. By comparison, two normal human glands were stimulated an average of 180%. Glands from three patients with secondary hyperplasia were also responsive to calcium. Thus, parathyroid glands from patients with primary and secondary hyperparathyroidism were characterized by a spectrum of responsiveness to calcium, and absolute "autonomy" was unusual. Even at high calcium concentrations, hormone release persisted at a low but definite level ("basal secretion"). The total number of functioning parathyroid cells is therefore a principal determinant in the oversecretion of PTH in hyperparathyroidism.


Hypoparathyroidism: a possible cause of osteomalacia .

A 17 year old man with longstanding hypocalcemia and hyperphosphatemia presented with incapacitating bone pain and progressive weakness nad bowing of the legs. The serum abnormalities were due to idiopathic hypoparathyroidism as evidenced by a decreased serum concentration of parathyroid hormone and an appropriate rise in urinary cyclic AMP and phosphate excretion, and serum calcium concentration, in response to exogenously administered parathyroid extract. The serum concentration of 1,25-dihydroxycholecalciferol was appropriately decreased. The bone findings were due to osteomalacia as documented by physical findings, bone roentgenograms, and bone biopsy. Normal renal tubular function, blood pH, and serum concentration of 25-hydroxycholecalciferol and elevated serum alkaline phosphatase excluded the common causes of osteomalacia. The data are consistent with the hypothsis that lack of parathyroid hormone causes both hypocalcemia and a decreased serum concentration of 1,25-dihydroxycholecalciferol which, in turn, limit the availability of calcium and cause defective synthesis of bone matrix resulting in abnormal mineralization.


Effect of simultaneous potassium and saline loading on plasma aldosterone levels.

To assess the relative importance of the renin-antiotensin system and potassium in the acute regulation of aldosterone secretion in the salt depleted state, normal subjects were infused with an asotonic saline solution containing potassium chloride (KCl) and the responses compared to the infusions of isotonic saline (500 ml/h) and KCl alone (20 mEq K+/h). Subjects were studied recumbent in balance on a 10 mEq sodium/100 mEq potassium diet. The rates of infusing saline and potassium were selected to produce comparable but opposite effects on the levels of plasma aldosterone. As plasma renin activity (PRA) declined and plama potassium increased during the combined saline-KCl infusions, plasma aldosterone levels appeared to be the arithmetic sum of the changes in these parameters. In fact, the calculated changes in aldosterone levels per unit change in PRA or potassium from each infusion alone, correlated well with the observed levels, further supporting an additive relationship. These data support the concept that in the salt depleted state the renin-angiotensin system and potassium are equally important in the acute regulation of plasma levels of aldosterone.


Plasma renin and angiotensin II measurement in hypertensive and normal subjects: correlation of basal and stimulated states.

Plasma renin activity (PRA) and angiotensin II (AII) were measured in peripheral venous samples obtained from the following subgroups: normal ambulant subjects (n = 61): normal subjects after 7 days treatment with a thiazide diuretic (n = 27): hypertensive subjects before and after a thiazide diruretic (n = 35); severely hypertensive subjects after diazoxide treatment (n = 14) and untreated severely hypertensive subjects (n = 27). In addition aortic (n = 27) and renal vein (n =48) samples were taken from the latter group during investigation of possible renovascular hypertension. Values for PRA and AII were clearly correlated in all groups. Further, calculated regression lines of PRA upon AII did not differ significantly from that calculated for normal subjects, with the exception of the intercept of the diazoxide treated group. However, when the regression line for renal venous samples was compared with that calculated for peripheral venous samples of all other subjects (199), there was a relative increase in the ratio of PRA to AII at all values. Thus, except in the renal vein, measurement of AII and PRA appear to reflect changes in activity of the renin-angiotensin system equally well and there is no support for the view that there is a discrepancy between measured AII and PRA in peripheral venous blood in the clinical situations examined here.


The simultaneous radioimmunoassay of seven steroids in human spermatic and peripheral venous blood.

Seven unconjugated neutral steroids, including testosterone and some of its precursors and metabolites, were measured in the peripheral and spermatic venous blood males, employing specific radioimmunoassays after the fractionation of steroids on Lipidex-5000 (hydroxyalkoxypropyl Sephadex) microcolumns. Respective mean concentrations (ng/ml) and ranges of steroids estimated in peripheral and spermatic venous blood in all groups of patients were as follows: pregnenolone, 0.71 (0.29-2.39) and 10.97 (0.83-30.1); progesterone, 0.31 (0.02-0.57) and 10.17 (1.51-33.24); 17 alpha-hydroxyprogesterone, 1.04 (0.48-2.20) and 37.33 (1.68-141.00); androstenedione, 1.01 (0.26-2.65) and 11.87 (0.97-30.18); testosterone, 3.84 (0.63-10.64) and 255.1 (2.85-619.1); 5alpha-dihydrotestosterone, 0.19 (0.07-0.28) and 3.74 (0.04-9.71); androsterone, 0.27 (0.12-0.47) and 0.97 (0.20-2.15). Concentrations are similar to those estimated by mass spectrometry and protein binding assays, except for androsterone which has not previously been measured in this context. The low, but significant testicular secretion of both 5alpha-hydrotestosterone and androsterone suggests that these two steroids are testicular androgen metabolites, and that androgen metabolism in this tissue may be monitored by way of their measurement in spermatic vein blood.


Hyperprolactinemia in girls with idiopathic precocious puberty under prolonged treatment with cyproterone acetate.

Pasma prolactin response to the acute injection of sulpiride (1.5 mg/k BW im) was measured at 0800-0900h in 4 girls with idiopathic precocious puberty before and after 6 to 11 months of continuous therapy with 50 mg daily of cyproterone acetate (CA) orally administered. Two additional girls were examined after 26 and 28 months of therapy, respectively. Mean baseline prolactin concentrations were significantly higher in untreated girls with precocious puberty as compared to that of normal controls of the same chronological age and of a comparable degree of sexual maturation (14.5 +/- 1.9 SE vs. 7.4 +/- 1.8 SE ng/ml and vs. 9.5 +/- 1.8 ng/ml, respectively; P less than .02). Treatment with CA caused a significant further increase of plasma prolactin concentration (P less than .02 as compared to pre-treatment values); no correlation was observed between prolactin concentration and duration of treatment. No significant change in the integrated areas of prlactin response to sulpiride occurred after prolonged CA therapy. The results suggest that in idiopathic precocious puberty the action of CA upon the hypothalamic-hypophyseal complex is not solely antigonadotropic and that prolactin secretion is enhanced in patients given this drug.


Thyroxine concentration in human milk.

Thyroxine (T4) concentrations were measured by RIA in 37 milk samples from 19 healthy eutryroid mothers, obtained between 3-165 days postpartum. The mean milk T4 concentration in the first week postpartum was 0.38 +/- 0.07 microgram/100 ml (mean +/- SEM). The mean T4 concentrations between 8 and 48 days rose to 4.27 +/- 0.50 microgram/100 ml (mean +/- SEM), and decreased to 1.11 +/- 0.25 microgram/100 ml (mean +/- SEM) after 50 days postpartum. The data suggest that human milk can provide a significant exogenous source of T4 to the infant. In the hypothyroid infant, the amount of T4 in human milk may delay clinical recognition of the disease. Although this exogenous source of T4 may alleviate the disease, it is insufficient to prevent the detrimetal effects of hypothyroidism.


Inhibition of the renin-aldosterone response to furosemide by indomethacin.

In 12 human volunteers, indomethacin was shown to inhibit renal prostaglandin produciton as reflected by RIA of urinary PGE2. The increases in plasma renin activity and plasma and urinary aldosterone following acute furosemide challenge were markedly blunted in the presence of indomethacin. In light of recent development indicating an intimate relationship between prostaglandins and renin release, these results provide further evidence for a pivotal role of renal prostaglandins in modulating the responsiveness of the renin-angiotensin-aldosterone axis in Mam.


Hypercalcemic hyperparathyroidism in hypophosphatemic rickets.

A 25-year-old white woman with sporadic hypophosphatemic rickets presented with a 7 year history of chronic mild hypercalcemia, osteitis fibrosa cystic and hypercalcemic nephropathy. Serum immunoreactive parathyroid hormone was elevated by greater than 100-fold and a 3.5 g parathyroid tumor was found at operation. Survey of the literature reveals that of 9 previous cases in which hypercalcemic hyperparathyroidism occurred in association with hypophosphatemic rickets, only two had classical x-linked familial hypophosphatemic rickets. It appears more than likely that this unusual combination of skeletal diseases represents the chance occurrence of primary hyperparathyroidism in patients with underlying x-linked familial hypophosphatemic rickets rather than a complication of phosphate therapy.


The influence of severe illness on gonadotropin secretion in the postmenopausal female.

Severe weight loss in amenorrheic premenopausal women may significantly depress gonadotropin secretion. Gonadotropin leves were studied in 111 postmenopausal women to determine if weight loss and cachexia could similarly affect gonadotropin function. Thirty-three healthy ambulatory postmenopausal women and twenty-seven healthy hospitalized women admitted electively were found to have a wide range of elevated values, whose mean did not differ significantly. Mean levels for both LH and FSH were significantly suppressed (P less than .005) in severely ill postmenopausal women both with or without weight loss. (formula: see text) Recovety from illness in six patients was associated with a rapid rise in FSH levels while LH remained depressed. Two severely ill patients studied over a 6 h period revealed constant depression of both gonadotropins without the characteristic pulses seen in four normal control postmenopausal women. These results indicate that gonadotropin secretion may be suppressed in severely ill postmenopausal women whether weight loss is present of not, and the central nervous regulatory mechanism responsible for intermittant release of gonadotropins is impaired. The disparate FSH and LH recovery indicates that if separate gonadotrophs exist, the LH gonadotroph is more severely affected than the FSH gonadotroph and/or that the responsible regulatory mechanisms are different.