High uptake of myo-inositol by rat pancreatic tissue in vitro stimulates secretion.

A recent study by Hokin-Neaverson, M., Sadeghian, K., Majumder, A.L., and Eisenberg, F. (1975) Biochem. Biophys. Res. Commun. 67, 1537-1544, demonstrates that free myo-inositol in the pancreas is significantly increased during intense cholinergic stimulation of secretion. Incubation of rat pancreatic tissue in medium with 100 mM myo-inositol increases 10-fold the endogenous content of free myo-inositol and elicits a prompt and sustained 50% increase in the rate of release of amylase activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis reveals that the electrophoretic pattern of the protein mixture released in the presence of 100 mM myo-inositol is the same as that of the secretory output released in the presence of 10 muM carbamylcholine. Microscopic examination of tissue pieces indicates that there is no significant decrease in zymogen granule content of the pancreatic acinar cells during incubation in medium with 100 mM myo-inositol. Jamieson, J.D., and Palade, G.E. (1967) J. Cell Biol. 34, 597-615, have shown that pulse-labeled secretory proteins in guinea pig pancreas first appear in zymogen granules 1 hour postpulse, becoming maximally accumulated in these storage sites by 2 hours postpulse. myo-Inositol (100 mM) stimulates release of pulse-labeled secretory proteins only if incubation in medium with 100 mM myo-inositol is initiated anytime during the first 80 min postpulse. The findings thus indicate that a high uptake of myo-inositol by rat pancreatic tissue in vitro selectively stimulates the release of just those secretory proteins being packaged in newly forming zymogen granules.


Formyl-methyl-methylenetetrahydrofolate synthetase-(combined). An ovine protein with multiple catalytic activities.

Formyltetrahydrofolate synthetase, methenyltetrahydrofolate cyclohydrolase, and methylenetetrahydrofolate dehydrogenase in sheep liver co-purify 200-fold to yield a homogeneous preparation containing a single protein species observed on discontinuous polyacrylamide gel electrophoresis in the presence of dodecyl sulfate. The synthetase and dehydrogenase activities migrate with the single protein band observed on polyacrylamide gel electrophoresis in two discontinuous buffer systems near pH 8. The protein sediments with a single symmetrical boundary with sedimentation coefficient (S20,w) = 8.30 S and elutes from molecular exclusion chromatography columns at a position corresponding to a diffusion coefficient (D20,w) = 3.99 X 10(-7) cm2 sec-1. Dodecyl sulfate-polyacrylamide gel electrophoresis of the protein intramolecularly cross-linked with dimethylsuberimidate shows one protein species of Mr = 218,000 in addition to the species of Mr = 108,500 characteristic of the unmodified protein. NH2-terminal analysis of the protein using 5-dimethylaminonaphthalene-1-sulfonyl (dansyl) chloride in the presence of dodecyl sulfate results in the recovery of a single dansyl amino acid, alanine. The three activities are thus shown to reside in a protein composed of two apparently identical subunits, and the trivial name formyl-methenyl-methylenetetrahydrofolate synthetase(combined) is proposed for this enzyme to suggest the multiple catalytic activities associated with the single protein species.


Fragmentation of the 95,000-dalton transmembrane polypeptide in human erythrocyte membranes.

The 95,000-dalton polypeptide in human red blood cell membranes constitutes about 25% of the membrane protein. Previous labeling studies have shown that different regions of this polypeptide are exposed to the inside and outside of the cell and have suggested a role for the protein in anion exchange across the membrane. This polypeptide has been fragmented by chymotrypsin digestion of intact red cells and by treatment of purified polypeptide with 2-nitro-5-thiocyanobenzoic acid, hydroxylamine, and N-bromosuccinimide. The sites of cleavage by each of these reagents have been located relative to the NH-2 and COOH-terminals of the intact 95,000-dalton polypeptide. Polypeptide obtained from cells labeled with 1-isothiocyanate-4-benzene [35S]sulfonic acid (an inhibitor of anion transport), 125I and lactoperoxidase, or 32P has been similarly fragmented and these labels have been assigned to specific regions of the polypeptide. There are at least two sites of phosphorylation of the polypeptide; the major sites lies within 10,000 daltons of the NH2-terminal requiring that this portion of the polypeptide lie inside the cell. Sites of chymotrypsin cleavage and 125I and lactoperoxidase labeling are in a 7,000-dalton region toward the COOH-terminal of the polypeptide; this region must lie outside the cell. Between these two regions the polypeptide must traverse the lipid bilayer an odd number of times. 1-Isothiocyanate-4-benzenesulfonic acid also labels the protein near the site of chymotrypsin cleavage.


Reversible inhibition of glucose transport in human erythrocytes by a series of pyridine derivatives.

A series of pyridine derivatives was chosen on the basis of structural similarity to glucose and to the hydrogen bonding capacity of this sugar. These compounds were found to reversibly inhibit glucose transport in a systematic fashion. The inhibition data can be rationalized by assuming that both competitive and noncompetitive binding sites exist on the cell membrane. The competitive site appears to have a much lower dissociation constant than the noncompetitive site for most derivatives. The effects of changes in substituents on the pyridine ring closely match, both in ring position and in the magnitude of the change in binding constant, those reported for derivatives of glucose. The stimulation of fluorodinitrobenzene inactivation of glucose transport by 2-amino-5-chloropyridine is in accord with this analysis. The strongest inhibitors bind about 100-fold more tightly than glucose itself. This series of pyridine compounds would appear to present a useful starting point for the development of site-specific irreversible inhibitors.


Effect of a supernatant protein on microsomal squalene epoxidase and 2,3-oxidosqualene-lanosterol cyclase.

Squalene epoxidation catalyzed by rat liver microsomes requires oxygen NADPH, and the 105,000 x g supernatant (S105). The supernatant can be replaced by a partially purified S105 protein (SPF) and phospholipids (Tai, H., and Bloch, K. (1972) J. Biol. Chem. 247, 3767). When washed microsomes are preincubated anaerobically with [14C]squalene and S105 without NADPH, followed by centrifugation and washing to remove the unbound squalene and S105, epoxidation in the presence of O2 and NADPH occurs subsequently at the same rate as in direct assays containing all required components from the start. Partially purified SPF (65-fold) shows the same effect. Washed microsomes preincubated anaerobically with squalene alone, or with bovine serum albumin instead of S105, also take up large amounts of squalene, but the squalene so incorporated is only poorly converted to epoxide. The epoxidation of endogenous squalene formed in liver homogenates from [14C]mevalonate is also stimulated by S105. The incorporation of squalene into microsomes is temperature dependent. 2,3-Oxidosqualene-lanosterol cyclase (cyclase) also requires S105 for optimal activity. It is suggested that the S105 protein acts internally within the microsomal membrane system facilitating the access of substrate to specific enzyme sites.


Control of mutation frequency by bacteriophage T4 DNA polymerase. I. The CB120 antimutator DNA polymerase is defective in strand displacement.

The ts CB1200 (antimutator) mutation in bacteriophage T4 DNA polymerase increases the accuracy of DNA replication since it results in a decrease in the frequency of mutations in other phage genes. The CB120 polymerases differs from the wild type enzyme in the slow rate at which it copies templates where primer extension requries displacement of polynucleotides base-paired to the template strand, even in the presence of the T4 DNA unwinding protein (gene 32-protein). The ratio of nucleotides turned over (DNA-dependent conversion of deoxynucleoside triphosphate to deoxynucleoside monophosphate) to nucleotides stably incorporated into product is 10 to 100 times higher with the mutant than wild type enzyme, depending on the DNA used as the template. This high turnover rate may increase the efficiency of removal of noncomplementary nucleotides by the antimutator enzyme and is in agreement with the findings of Muzyczka et al, (Muzyczka, N., Poland, R. L., and Bessman, M. J. (1972) J. Biol, Cehm. 247, 7116-7122) with the L141 and L42 antimutator T4 DNA polymerases. Since the 3'- to 5'-exonuclease activity of the CB120 mutant polymerase is not higher than that of the wild type enzyme, it is suggested that the high turnover rate may result from increased opportunity to remove newly incorporated nucleotides due to the slow rate at which the mutant enzyme moves to the next template nucleotide. In the accompanying paper we show that the CB120 antimutator polymerase also initially selects incorrect nucleotides for incorporation less frequently than the wild type enzyme. Thus this antimutator polymerase appears to have both greater accuracy in nucleotide selection and an enhanced ability to remove incorrect nucleotides.


Control of mutation frequency by bacteriophage T4 DNA polymerase. II. Accuracy of nucleotide selection by the L88 mutator, CB120 antimutator, and wild type phage T4 DNA polymerases.

The accuracy of nucleotide selection by wild type, L88 mutator, and CB120 antimutator T4 DNA polymerases has been compared by measuring both stable incorporation of complementary and noncomplementary nucleotides into polymer and the DNA-dependent conversion of deoxynucleoside triphosphate to monophosphate. The increased accuracy of the CB120 antimutator enzyme is shown by a ratio of utilization of incorrect to correct nucleotides with poly(dA)-poly(dT) as template which is only 10 to 30% of that of the wild type enzyme. In contrast, the ratio of incorrect to correct nucleotide utilized by the L88 mutator enzyme with this template was higher than that of the wild type enzyme, in agreement with the report of Hershfield (Hershfield, M.S. (1973) J. Biol, Chem. 248, 1417-1423). The antimutator, mutator, and wild type enzymes each have a much higher apparent Km for the noncomplementary nucleotides than for complementary nucleotides with this template. The L88 mutator polymerase has a higher "Km" for poly(dA)-poly(dT) than the wild type enzyme in reactions with both complementary and noncomplementary nucleotides. The antimutator polymerase has an elevated "Km" for polymer only for reactions with noncomplementary nucleotides. The wild type and L88 mutator enzymes also utilized both noncomplementary nucleotides much more frequently than the CB120 antimutator enzymes with poly [d(A-T)] as the template-primer. The T4 gene 32DNA unwinding protein appears to facilitate the correct reading of this template since it decreases the ratio of incorrect nucleotides utilized by both the wild type and L88 mutator polymerases.


Alpha-Hydroxylation of lignoceric and nervonic acids in the brain. Effects of altered thyroid function on postnatal development of the hydroxylase activity.

Rat brain postnuclear preparations catalyzed the alpha-hydroxylation of nervonic acid with an apparent Km of 3 muM. Evidence has been presented which suggests that nervonic acid in the brain is hydroxylated by the same enzyme system which hydroxylates lignoceric acid. The hydroxylase activity in brains of normal (euthyroid) rats increased rapidly from a low in the period immediately following birth to a maximum at the 23rd day and then declined to a low level characteristic of the mature brain. Neonatal hypothyroidism retarded the development of the activity and shifted its peak to the 39th day after birth. Conversely, neonatal hyperthyroidism accelerated the entire developmental pattern and shifted the peak to the 16th day after birth. The hydroxylase activity in mouse brain was also increased by thyroid hormone administration from the 13th through the 18th day after birth. Unlike normal mice, the low activity in jimpy mice was not affected by this treatment. It is concluded that thyroid hormones play an important role in the control of brain fatty acid alpha-hydroxylation. The stimulation of alpha-hydroxy fatty acid synthesis in response to hyperthyroidism during the early postnatal period may be one of the major effects of thyroid hormones in accelerating myelination of the central nervous system.


On the mechanism of regulation of omega oxidation of fatty acids.

The stimulatory effect of starvation on omega oxidation of stearate by the 20,000 X g supernatant fluid of rat liver homogenates was studied. The effect was obtained after starvation for 24 hours. Starvation for longer times did not further increase omega oxidation. The stimulatory effect of starvation on omega oxidation of stearic acid was accompanied by a reduced incorporation of stearic acid into phosphatidic acid, diglycerides, and triglycerides. Substitution of the 100,000 X g supernatant fluid from liver homogenate of starved rats with 100,000 X g supernatant fluid from liver homogenates of control rats reduced the microsomal omega oxidation of stearic acid with a simultaneous increase in incorporation of stearic acid into the different glycerides. Under the latter conditions almost no free stearic acid could be isolated from the incubation mixture after the incubation. Of three different soluble factors necessary for glyceride formation, ATP appeared to be the most important from a regulatory point of view. Thus the soluble fraction of liver homogenate from a starved rat was shown to contain suboptimal concentrations of ATP. Addition of physiological amounts of ATP to the 20,000 X g supernatant fluid of homogenate of liver of starved rats had the same effect as addition of 100, 000 X g supernatant fluid from liver homogenate of control rats, i.e. decrease in omega oxidation and increase in formation of glycerides. Addition of sn-glycerol 3-phosphate and CoA-SH in amounts optimal for glyceride formation to the 20,000 X g supernatant fluid of liver homogenate of starved rats had only small effects on omega oxidation and glyceride formation. The results are consistent with a competition for free fatty acids between the acyl-CoA synthetases involved in biosynthesis of glycerides and the microsomal hydroxylase(s) involved in omega oxidation of fatty acids. The concentration of ATP in the soluble fraction is of importance in this competition. The possibility is discussed that this competition is of importance also under in vivo conditions and that a decreased rate of esterification in the starved state is responsible for the higher excretion of omega-oxidized fatty acids in urine in the ketotic state.


Modification of the 5' terminus of mRNA by an RNA (guanine-7-)-methyltransferase from HeLa cells.

The 5' termini of many viral and cellular mRNAs contain sequences of the type m7G(5")pppNm. An RNA (guanine-7-)-methyltransferase that specifically methylates the 5'-terminal guanosine residue of RNAs ending in the dinucleoside triphosphate G(5')pppN- has been purified from the cytoplasm of HeLa cells. Approximately two-thirds of the methyltransferase activity detected in an assay employing umnethylated vaccinia virus mRNA as acceptor was located in the cytoplasm when cells were disrupted by Dounce homogenization; 30% of the cytoplasmic activity was associated with ribosomes but was removed by washing with 0.5 M KCl. The enzyme was purified 165-fold from the cytoplasm by removing nucleic acid by phase partition followed by ammonium sulfate precipitation and column chromatography on DEAE-cellulose, denatured DNA-agarose, and CM-Sephadex. The partially purified enzyme preparation methylated heterologous tRNAs as well as vaccinia mRNA, but the tRNA methyltransferases could be separated from the mRNA activity by sucrose gradient sedimentation and gel filtration on Sephadex G-200. The product of the partially purified enzyme using vaccinia mRNA as substrate was exclusively 7-methylguanosine located in the terminal dinucleoside triphosphate. In addition to RNAs and synthetic polyribonucleotides terminating in a dinucleoside triphosphate, free G(5')pppG could be methylated but GTP, GDP, and G(5')pppG could not. The enzyme also methylated the dinucleoside diphosphate G(5')pppG but much less efficiently than G(5')pppG. An S20, W of 3.8, a Stokes radius of 3.6 nm, and a molecular weight of 56,000 were obtained from sucrose gradient sedimentation and Sephadex G-200 column chromatography.


Carbohydrate structure of glycopeptides isolated from an hepatic membrane-binding protein specific for asialoglycoproteins.

Two variant glycopeptide constituents of an hepatic membrane protein responsible for the clearance of circulating asialoglycoproteins have been isolated and characterized. Their relative abundance has been estimated to be 14 and 2 mol, respectively, per mol of intact protein. The larger component, glycopeptide I, is comprised of sialic acid, galactose, mannose, glucosamine, and aspartic acid in a molar ratio of 3:3:2:5:1. The smaller neutral fraction contains only two sugars and the molar relationship of mannose, glucosamine, and aspartic acid is 8:2:1. Based on the results of sequential digestion with specific glycosidases and periodate degradation, a carbohydrate sequence for both glycopeptides is proposed.


Deacetylation reaction catalyzed by Salmonella phage c341 and its baseplate parts.

Phage c341 was shown to cleave off 14C-acetyl groups from a 14C-acetylated polysaccharide, prepared by the enzymatic acetylation of alkali-treated Salmonella anatum O-polysaccharide catalyzed by a membrane fraction from S. anatum (O-10 transacetylase). No deacetylation reaction was detected if the substate was prepared from alkali-treated O-polysaccharide of Salmonella newington. The structural difference of these O-polysaccharides is the anomeric configuration of the linkage between the common mannosyl-rhamnosyl-galactose repeating units. A soluble protein obtained from phage c341 lysate, which has previously been identified as the free form of the baseplate parts of this phage, showed the deacetylase activity, a result indicating that the baseplate is responsible for the enzymatic activity of the phage particles. These results suggest that as in the case of other Salmonella phages such as epsilon15, epsilon34, and P22 that contain glycosidases as baseplates, the baseplate deacetylase of c341 plays a role for phage adsorption through the formation of enzyme-substrate type complexes with the receptor O-polysaccharide.


Physical properties of antitumor glutaminase-asparaginase from Pseudomonas 7A.

Glutaminase-asparaginase from Pseudomonas 7A appears to have four subunits with a molecular weight of 36,000 +/- 500 by sedimentation equilibrium in 5.9 M guanidine HCl and 34,000 by amino acid analysis. Analytic sedimentation equilibrium of the native enzyme showed a molecular weight of 140,000 +/- 3,300 with no signs of association or dissociation. Moving boundary and zone sedimentation in buffer showed normal behavior with sedimentation coefficients of 7.92 and 7.75 S, respectively. In contrast, the enzyme appeared to polymerize during zone sedimentation when the initial protein concentration was greater than 1 mg/ml and the buffer contained asparagine, glutamine, or 5-diazo-4-oxonorvaline. An extension of our method for active enzyme sedimentation is described which utilizes the changes in absorption during hydrolysis of asparagine by low concentrations of enzyme. Polymerization was not seen under these conditions.


Interaction of human interferons with immobilized hydrophobic amino acids and dipeptides.

Human fibroblast interferon binds to L-tryptophan, D-tryptophan, L-phenylalanine, and L-tyrosine, all immobilized directly to cyanogen bromide-activated agarose, as well as to L-tryptophan and D-tryptophan methyl ester, both immobilized via molecular arms. The retention of fibroblast interferon is selective and results in a 2300-fold purification. Human leukocyte interferon binds neither to L-tryptophan attached directly to an agarose matrix nor to L-tryptophan immobilized via a molecular arm; it binds, however, to immobilized L-tryptophyl-L-tryptophan and L-tryptophyl-L-tryrosine. When retained, both interferons cannot be displaced unless ethylene glycol is included in the eluant, indicating a hydrophobic interaction. The interaction takes place under physiologic solvent conditions, thus revealing the high intrinsic hydrophobicity of both interferons.


Chemical properties of two antigens controlled by the major histocompatibility complex of the chicken.

The chemistry of antigens controlled by the major histocompatibility complex of the chicken has been investigated. Peripheral blood leukocytes of five chicken strains were radioactively labeled by incorporation of tritiated amino acids, enzymatic iodination, or mild periodate oxidation followed by reduction with tritiated sodium borohydride. Membrane-bound antigens were solubilized with Nonidet P-40 and purified by a two-step immunoprecipitation procedure. The resulting immunoprecipitates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing or nonreducing conditions. The chicken major histocompatibility antigens (B antigens) are composed of polypeptide chains with molecular weights (in the reduced form) of 40,000 to 43,000 and 11,000 to 12,000, as deduced from their mobilities in internally calibrated gels. The larger chains, when isolated from different chicken strains, can have slightly different mobilities in such gels; thus the mobilities of the large polypeptides from strains WA and WB correspond to molecular weights of 40,000 and 43,000, respectively. Gels run under nonreducing conditions give similar results; in particular no dimers or oligomers of the larger chains are detectable, thus ruling out an immunoglobulin-like structure for the B antigens. The large chains are labeled by the periodate oxidation-reduction procedure, suggesting that they are glycoproteins (probably containing sialic acid), while the small polypeptides cannot be labeled by this method. An additional protein or proteins of apparent molecular weight about 30,000 could be precipitated from 3H-amino-acid-labeled leukocyte lysates by one particular anti-B serum. This protein(s), named B-L, differs from the B major histocompatibility antigens in that it is not associated with the small chain of molecular weight 11,000 to 12,000 and does not occur on erythrocytes. Thus, at least two protein chains are coded for (or have their synthesis controlled by) genes in the chicken major histocompatibility complex.


Polycentric total knee arthroplasty. A two-year follow-up study.

Polycentric total knee arthroplasty provided significant relief of pain in 86 per cent of 500 knees. The independence and activity levels of the patients increased dramatically. The frequency of major complications as reflected by reoperation was 10 per cent in this series. There was a 2.8 per cent deep infection rate. One-third of the infected knees were salvaged and two-thirds required arthrodesis. Loosening of a component was noted in 2.4 per cent. After operation the average range of motion was from 6 to 101 degrees of flexion, for a range of 95 degrees; this was a 5-degree increase over average preoperative motion. Ninety-six per cent of the patients expressed satisfaction with the surgical result.


Geometric total knee arthroplasty. A two-year follow-up study.

In a study of our experience with the first 119 geometric arthroplasties of the knee done at the Mayo Clinic, reported after a minimum follow-up period of two years, we noted that relief of pain was the primary goal in 84 per cent of the cases and this was achieved in the majority. The average total motion was relatively unchanged but it became more useful postoperatively.


A comparison of four models of total knee-replacement prostheses.

Twenty-nine knees with unicondylar, sixty-four with duocondylar, fifty with Guepar, and fifty with geometric prostheses were studied. The follow-up ranged from two to three and one-half years. The unicondylar prosthesis was used in the mildest cases and gave the least complications, but the quality of results was not superior to that achieved with the other prostheses. The duocondylar model was best suited for knees with rheumatoid arthritis and mild deformity. The geometric prosthesis was the best condylar prosthesis for osteoarthritis with moderate to severe deformity, but gave the worst results in knees with rheumatoid arthritis. The Guepar prosthesis was used in the worst knees and gave the best results, but it had the highest infection rate and was the most difficult to salvage. A radiolucency was observed in about 60 per cent of the condylar replacements around the tibial component and in 45 per cent of the Geupar replacements around the femoral component. The significance of this cannot yet be determined but it suggest that the fixation may not be ideal. In all types, residual pain was most frequently attributed to the patellar compartment. Patellectomy was not a solution.


Modular total knee-replacement arthroplasty. A review of eighty-nine patients.

Fifty-eight osteoarthritic and thirty-one rheumatoid patients underwent modular total knee-replacement arthroplasty. The major indication for the operation was relief of pain. Contraindications to this resurfacing arthroplasty included varus-valgus instability of over 20 degrees, combined varus-valgus instability with flexion contracture of over 40 degrees, marked recurvatum, and predominant patellofemoral symptoms. In 59 per cent of the osteoarthritic and 58 per cent of the rheumatoid patients, complete relief of pain was evident when they were evaluated twenty-four months after surgery, while another 35 per cent of each group had only mild pain related to inclement weather. Their ability to walk long distances without support or limp was increased. Range of motion and ability to climb stairs were not significantly improved.


Failed polycentric total knee prostheses.

Four failed polycentric total knee prostheses were evaluated by clinical examination of the patients and surface analysis of the removed prosthetic components. The failures were secondary to a poor relationship of the prosthetic components in three cases and to abutment of the tibial spine against the femoral condyle in the fourth. A poor relationship of the prosthetic components was not necessarily secondary to malpositioning, because the positioning may have been accurate but the relationships became abnormal when the knee was unstable. Scanning electron-microscope examination revealed various wear patterns but no significant correlation was noted between the morphological type and the degree of wear. Accurate placement of the components and restoration of knee stability were critical requirements for the polycentric total knee prosthesis to be successful.


Patterns of movement of totally intrinsic-minus fingers based on a study of one hundred and forty-one fingers.

A detailed study of metacarpophalangeal flexion and interphalangeal extension movements of 141 fingers with complete intrinsic-muscle paralysis due to leprosy showed that long flexors and long extensors produce movement at the metacarpophalangeal and proximal interphalangeal joints simultaneously, and not successively as is generally believed. The amounts of flexion resulting from long flexor activity are almost equal at the two joints and metacarpophalangeal flexion is achieved without excessive flexion of the proximal interphalangeal joint, but this is masked by the claw-finger deformity. The movement resulting from activity of the long extensor is complex and there are three or more qualitatively different patterns of extension. Although the long extensor produces simultaneous extension at the metacarpophalangeal and proximal interphalangeal joints, the latter consistently lags behind the former so that full extension is not achieved at the proximal interphalangeal joint even when the metacarpophalangeal joint is maximally extended. The diverse patterns of extension are not related to duration of degree of clawing or to any particular finger.


Evaluation of the two-stage flexor-tendon reconstruction in severely damaged digits.

From July 1971 to July 1974, the two-stage tendon-grafting procedure of Hunter and Salisbury was performed in thirty-two severely damaged digits in twenty-five patients. Of these digits, twenty-three fingers and five thumbs could be evaluated for gain in total active flexion (expressed as per cent of preoperative passive flexion) and for gain in total active motion (expressed as per cent of total preoperative passive motion) after follow-ups ranging from six to fifty months. The results for total active flexion were 60.9 per cent good, 21.7 per cent fair, and 17.4 per cent poor, and for total active motion 21.7 per cent good, 56.5 per cent fair, and 21.7 per cent poor. Complications were frequent after both stages and included infection, migration of the rod, and adhesions within the proximal end of the newly formed sheath. Flexion contractures were a significant problem.


Late reconstruction of the ligaments following acromioclavicular separation.

Complete separation of the acromion from the clavicle in nine young patients was repaired by a new technique with use of a tendon graft. The operation was performed, on average, twenty-nine weeks after the injury. After surgery seven patients were enabled to engage in strenuous sports or construction work and the other two to return to daily activities with occasional heavy lifting.


Compensating osteotomy at the base of the femoral neck for slipped capital femoral epiphysis.

A compensating osteotomy for correction of the varus and retroversion components of severe slipped capital femoral epiphysis is presented. It is safer than a cuneiform osteotomy done through the callus of the slipped epiphysis because the line of osteotomy is distal to the major blood supply of the posterior retinaculum. The threaded Steinmann pins used for fixation sross both the osteotomy site and the epiphyseal plate, and therefore not only are the mechanical relationships restored, but further slipping is prevented also.


Stiffness of the proximal interphalangeal joints in rheumatoid arthritis. The role of flexor tenosynovitis.

In nineteen hands (seventy-four fingers) of eleven women and one man with rheumatoid arthritis there was restriction of active and passive motion of the proximal interphalangeal joints, with signs of flexor tenosynovitis but no clinical or roentgenographic evidence of involvement of the joint. The nineteen hands were treated by flexor tenosynovectomy (palm only in nine, palm and carpal tunnel in five, both palm and digits in four, and digit, palm, and wrist in one) combined with manipulation of the joint under regional anesthesia. After an average follow-up of twenty-one months (range, six to thirty-six months), the average range of active motion had increased from 40 to 84 degrees and the average range of passive motions, from 57 to 87 degrees. Only three patients had unsatisfactory results, one because of persistent unexplained swelling and two because of recurrence of the tenosynovitis.


Milwaukee brace correction of idiopathic scoliosis. A biomechanical analysis and a restrospective study.

We analyzed the biomechanics of Milwaukee brace treatment of idiopathic scoliotic patients through simulation in five computer-constructed model spines. The contributions to correction of each component of the brace were examined in these model spines, and some of the mechanical principles that determine the outcome of brace treatment were studied. The validity of the stimulation findings was then tested by a retrospective analysis. Simulation was used to predict the outcome of milwaukee brace treatment in sixty-eight patients. In 81 per cent of these patients, the actual outcome agreed with the prediction. The study suggests that careful adherence to mechanical principles in the use of a Milwaukee brace will result in successful treatment of more patients.


Tenderness at motor points. A diagnostic and prognostic aid for low-back injury.

In patients with low-back injury the motor points of some muscles may be tender. Of fifty patients with low-back "strain", twenty-six had tender motor points and twenty-four did not, while forty-nine of fifty patients with radicular signs and symptoms suggesting disc involvement had tender motor points, and the one without such tender points had a hamstring contusion which limited straight leg raising. Of fifty controls with no back disability, only seven had mild tender points after strenuous activity, while forty-six of another fifty controls with occasional back discomfort had mild motor-point tenderness. In all instances the tender motor points were located in the myotomes corresponding to the probable segmental levels of spinal injury and of root involvement, when present. Patients with low-back strain and no tender motor points were disabled for an average of 6.9 weeks, while those with the same diagnosis but tender motor points were disabled for an average of 19.7 weeks, or almost as long as the patients with signs of radicular involvement, who were disabled for an average of 25.7 weeks. Tender motor points may therefore be of diagnostic and prognostic value, serving as sensitive localizers of radicular involvement and differentiating a simple mechanical low-back strain from one with neural involvement.


Total hip reconstruction in chronically dislocated hips.

The pathological anatomy of chronically dislocated hips makes reconstruction for more difficult in them than in most cases. The acetabular component must be seated at the site of the original triradiate cartilage and the femur must be shortened four or more centimeters to prevent excessive limb lengthening. This means that the femoral component must be seated in the smallest, strightest portion of the intramedullary canal. A specially designed prosthesis is often needed. Twenty-two hips were operated on in this series, and all patients had relief of pain and improvement of gait. One major complication occured: a sciatic-nerve palsy due to overlengthening of the femur and improper postoperative positioning.


Pyogenic arthritis of the sacro-iliac joint. Long-term follow-up.

Nine cases of sacro-iliac pyarthrosis are presented. The difficulty in localizing the infection is attributable mostly to failure to appreciate the posteriorly situated physical findings. This, and the difficulty with early roentgenographic demonstration of the lesion, may lead to unnecessary abdominal exploration (as in two of our patients) or to prolonged delay in diagnosis and hence spread of the infection. Awareness of the usual physical findings and prompt use of radioisotope scanning to localize the infection led to earlier diagnosis and avoidance of surgery in three patients seen recently. Antibiotic therapy, with or without surgery, led to cure in all patients, with minimum sequelae.


The use of the Zuelzer hook plate in fixation of olecranon fractures.

Twenty-five cases of fracture of the olecranon process were treated using a modification of the Zuelzer hook plate, and uniformly excellent or good results were obtained in all cases in which there were no other associated traumatic lesions about the elbow joint. Regardless of the degree of comminution of the fracture, obliquity of the fracture line, or age of the patient, almost a full range of motion was obtained in every case. No failure of the appliance or loss of position occurred in any case despite the fact that active motion was usually instituted in one to two weeks.


An intramedullary fixation device for the proximal part of the femur. Nine years' experience.

Nine years' experience with a combined intramedullary rod and triflanged nail in eighty-four non-pathological fractures in the subtrochanteric region of the femur is reported and a system of classifying these fractures based on morphology is offered. The appliance was successful in permitting early mobilization of patients and afforded a high rate of union of the fractures. The problems of varus displacement of the distal fragment and protrusion of the device into the joint were not encountered, and there was only one mechanical failure.


Primary synovial chondromatosis of the ankle. A case report.

A case of primary diffuse synovial chondromatosis of the ankle with long-term follow-up is reported. Trauma is implicated as a precipitating factor. This unusual, distinctive synovial neoplasm presents readily recognizable pathological features (Fig. 2). Specific diagnostic criteria are presented. The differential diagnosis of primary synovial chondromatosis includes secondary reactive synovial chondromatosis and synovial chondrosarcoma. The usual sources of osteochondritic loose bodies, on the other hand, are traumatic joint disruption, osteoarthritis, and infectious disorders. Acute primary synovial chondromatosis is adequately treated by removal of the loose bodies and synovectomy. Postoperative prognosis is dependent on the stage of the disease at the time of surgery. In this acute case, the patient had minimum impairment, and ankle roentgenograms twenty years after the arthrotomy were normal (Fig. 3).


The results of early treatment of typical congenital dislocation of the hip in Malmö.

The late results of early treatment of congenital dislocation of the hip in the von Rosen splint have been recorded in a follow-up study of 111 children with an average age of ten years (range eight to sixteen). The clinical diagnosis had been made within two days of birth and in most cases confirmed by radiological examination. Splintage was in all cases begun within the first few days and maintained for an average period of ten weeks. Two cases of failure of the initial treatment were encountered but had good final results. The follow-up study included a radiological examination in which were recorded the general configuration of the joint, the CE-angle described by Wiberg, the presence or absence of an os acetabuli and the roundness of the femoral head. The findings were compared with those in a control group of 222 normal children. All the treated hips developed normally except for a single case of minor dysplasia and a single case of slight enlargement of the femoral head and neck following appearances suggestive of avascular necrosis at the age of two years.


Patello-femoral joint mechanics and pathology. 1. Functional anatomy of the patello-femoral joint.

Cadaver knee joints were mounted so that life-like forces of weight-bearing were simulated. The patello-femoral contact areas were defined under load throughout the range of movement by the dye method. During movement from extension to 90 degrees of flexion a band of contact sweeps across the patella from inferior to superior pole, but the odd facet makes no contact. At about 135 degrees of flexion separate medial and lateral contact areas form, the medial one limited to the odd facet. From extension to 90 degrees of flexion the patella holds the quadriceps tendon away from the femur, but in further degrees of flexion an extensive "tendo-femoral" contact area forms. Between 90 degrees and 135 degrees of flexion the patella rotates and the ridge between the medial and odd facets engages the femoral condyle. The odd facet is shown to be a habitual non-contact area and the ridge to be subject to high load, observations which correlate with cartilage lesions described in Part 2 of the paper.


Patello-femoral joint mechanics and pathology. 2. Chondromalacia patellae.

Two distinct lesions affect the articular cartilage of the patella. Surface degeneration occurs particularly on the odd facet; it is age dependent, often present in youth and it becomes more frequent with increasing age. It probably does not occasion patello-femoral pain in youth, but may predispose to degenerative arthritis in that joint in later years and is regarded as a consequence of habitual disuse. The term "basal degeneration" is used to describe a lesion in which there is a fasciculation of collagen in the middle and deep zones of cartilage without, at first, affecting the surface. It was found astride the ridge separating the medial from the odd facet in twenty-three adolescents who had complained of prolonged patello-femoral pain. They were treated by excision of the disc of affected cartilage, with relief of pain in most cases. The pathogenesis of basal degeneration is related to the functional anatomy of the patella.


The Shiers arthroplasty of the knee.

Thirty-one patients with rheumatoid arthritis were reviewed after Shiers arthroplasty of the knee joint for which the main indication was intractable pain. Seven patients had the operation done to both knees. This review was done to assess the long-term results two to seven years later. All patients were clinically and radiologically assessed, and our results showed that pain and instability recurred within eighteen months. Thus it is likely that a prosthesis which allows rotation, and in which the bearing surfaces are metal on plastic, will replace the Shiers prosthesis.


Pseudoarthrosis after spinal fusion for scoliosis.

A series of 246 patients with scoliosis and attempted fusion had exploration performed six months later in order to detect and treat any pseudarthrosis at an early stage and so prevent subsequent loss of correction. Bilateral or unilateral pseudarthroses occurred in 25 per cent and were of three types--definite, hairline and doubtful. Single unilateral pseudarthroses accounted for 6 per cent and were of little if any clinical significance. The hairline pseudarthroses could not be seen radiologically and were easily missed at exploration. In general the pseudarthroses were least common in the more rigid parts of the spine and in curves which by nature of their aetiology or long duration had become most rigid. Neither the initial severity of the curve nor the degree of correction obtained before the initial attempted fusion had any apparent effect on the incidence. Follow-up for an average of four years has shown that a pseudarthrosis is of little significance with regard to the ultimate result provided it is recognised early and repaired.


Steroid-induced avascular necrosis of the head of the humerus. Natural history and management.

Ninety-five patients with steroid-induced avascular necrosis of bone have been personally treated by the author. Of these, eighteen had a lesion of the head of the humerus, on one or both sides. The conditions for which the steroids were given included post-transplantation, lupus erythematosus, glomerulonephritis and asthma. The characteristic lesion began as a subchondral osteolytic area which frequently progressed to collapse. The articular cartilage divided from the subchondral bone, either becoming detached as a free cap or at a later stage reattaching. In some cases the lesion was minimal and the symptoms were slight. Conservative treatment has consisted of pendulum exercises and avoidance of abduction, particularly against resistance. In fourteen patients this led to satisfactory function with only intermittent symptoms. Four patients required replacement of five humeral heads with Neer's prostheses. After one to seven years the results of all five were classified as excellent in terms of absence of symptoms and a free range of movement.


Anterior cervical subluxation.

Anterior subluxation of the cervical spine from flexion-rotational violence is often overlooked. Either radiography is omitted or the slight displacement visible on the films is not recognised. A method for recognising these injuries is suggested. Five cases are described showing that in these subluxations the soft-tissue damage is severe, that late increasing displacement can occur, with varying neurological sequelae, and that instability can persist indefinitely. Four of the patients required stabilisation by operation, in contrast with complete dislocations in which 80 to 90 per cent fuse spontaneously.


Hidden flexion injury of the cervical spine.

This paper describes seven patients who developed late vertebral deformity after flexion injuries of the cervical spine. In four the clinical and radiological features were subtle and because the patients walked into an emergency department the severity of the injury was not initially appreciated. Certain specific clinical and radiological features of flexion injury are described and emphasis is placed on the importance of correct management. A radiological tetrad is described which should alert the surgeon to the possibility of damage to the posterior interspinous complex of the cervical spine and so lead to further radiological investigations. Despite the frequency of flexion injuries the alarming complications described in this paper are rare.


Seat belt fractures of the cervical spine.

Three unusual injuries of the cervical spine in wearers of safety belts are reported. In each case, the presence of diagonal bruising and abrasions in the line of the sash strap indicated its involvement in the mechanism of the injury.


Legg-Calvé-Perthes' disease. Morphological studies in two cases examined at necropsy.

Specimens of femoral heads were studied at necropsy in two cases of Legg-Calve-Perthes' disease. One was that of a boy aged four years ten months who died from appendicitis; the other was from a boy aged six years who died from a malignant glioma. Both had been treated for one and a half years for Legg-Calve-Perthes' disease which was in a stage of repair at the time of death. The diseased femoral heads were moderately flattened but the surface cartilage was intact. Epiphysial bone and bone marrow were partly replaced by cartilage, fibrous tissue and granulation tissue, and new bone was being formed. Inflammatory reaction was inconspicuous. Enchondral bone formation was only slightly decreased, and the structure of the growth plate was undisturbed. There was no sign of systemic bone disease. In the first case the changes indicated that more than one episode of ischaemia had occurred, and an occlusion--probably from an old thrombus--was demonstrated in the posterior inferior retinacular artery of the femoral head. The last episode of ischaemia, furthermore, had caused infarction of part of the metaphysial bone. In both cases, the central area of the metaphysial bone of the affected femur contained fat, but there were few haemopoietic cells and it therefore looked pale. The findings are discussed in relation to previous work on the pathology in Legg-Calve-Perthes' disease, recent information on the vascularisation of the femoral head in children, and experimental and comparative animal studies.


Overgrowth of the femur after fracture of the shaft in childhood.

Twenty-six children conservatively treated for fracture of the femoral shaft have been reviewed with regard to differences in limb length seven to ten years after the injury. In nine patients the clinical measurements were checked against radiographic measurements of both femora and tibiae. The following conclusions were reached. Nearly two-thirds of the patients had overgrowth of the femur of 10 millimetres or more. Shortening of 15 to 20 millimetres at the fracture site was well compensated for by accelerated growth. Growth acceleration seemed to take place during the healing period and the difference at the end of healing was permanent. Overgrowth was promoted by comminuted and long oblique fractures and by overriding of the fracture ends, but was not influenced by the age at fracture, the duration of treatment or the level of fracture of the shaft. Growth of the tibia was not affected by the femoral fracture.


Parastremmatic dwarfism.

A girl aged ten, of Cape Coloured stock, with typical features of parastremmatic dwarfism has been investigated. The clinical manifestations included disproportionate dwarfism, limb deformity, a short stiff neck, and marked thoracic kyphosis. The radiographic changes were dramatic, the skeleton having a "flocky" appearance due to patches of radiolucency in an irregular lattice of sclerosis. The metaphyses and epiphyses of the long bones were grossly expanded, and the vertebrae were flattened and distorted. The clinical and radiographic features of five other previously reported individuals with parastremmatic dwarfism were very similar to those of our patient. The differential diagnosis of this condition includes metatrophic dwarfism, diatrophic dwarfism and the spondylo-epiphysial dysplasias. However, the unique "flocky" radiographic appearance of the bones permits diagnostic precision. There is some evidence to indicate that parastremmatic dwarfism might be transmitted as an autosomal dominant, although this is by no means certain. The fact that our patient had seven normal siblings and unaffected parents would be compatible woth autosomal recessive inheritance.


Massive osteolysis complicated by chylothorax successfully treated by pleurodesis.

A rare complication of massive osteolysis (disappearing bones, Gorham's disease)is presented--namely involvement and invasion of the thoracic duct by the angiomatous tissue spreading locally from involved upper thoracic vertebrae. The resultant chylothorax in our patient was investigated and the site of the lymphatic fistula established by lymphography. Operative obliteration of the chylous leak was successfully achieved and the patient has survived. A review of the literature shows this complication to have been fatal in six other cases, including Gorham's first patient.


Madelung's disease of the wrist and dyschondrosteosis.

Twenty-six index patients with primary Madelung's deformity of the wrist were examined, as were sixty-five of their relatives. Standard radiographs were taken and measured to determine whether mesomelic dwarfism, a feature of dyschondrosteosis, was present. No case of mesomelia was found. No evidence of Madelung's deformity was found in the relatives of the index patients, who were all female. It is concluded that primary Madelung's deformity of the wrist is not an expressions of dyschondrosteosis, that it is not inherited, and that it does not occur in the male. One index patient with dyschondrosteosis was examined. Eight of her relatives were similarly affected, and three of these were male. It is suggested that previously recorded male cases of Madelung's deformity of the wrist were expressions of dyschondrosteosis or of secondary deformity.


A radiological sign of entrapment of the median nerve in the elbow joint after posterior dislocation. A report of two cases.

Two boys with entrapment of the median nerve in the elbow joint after closed reduction of a posterior dislocation with fracture of the medial epicondyle showed a characteristic radiological sign in the anteroposterior radiograph after two to three months. The sign was a depression in the cortex on the ulnar side of the distal humeral metaphysis, with interruption of the local periosteal reaction. At operation in both patients the depression was found to correspond with the place where the median nerve reached the posterior surface of the humerus. Radiographs taken after transverse section of the nerve above and below the joint capsule and end-to-end suture showed gradual disappearance of the cortical depression.


The control of pain after Keller's operation by the instillation of local anaesthetic before closure. A prospective controlled trial.

With the aim of controlling pain after Keller's operation, a solution of a long-acting local anaesthetic was instilled into the field before closure in a series of thirty patients. In a controlled prospective trial this simple technique by itself proved to be significantly better than either foot block at the ankle or the usual analgesics and was free from local or systemic complications.


Waldeyer's hernia.

One of the rare sites of internal abdominal hernia is Waldeyer's peritoneal fossa. A case report in a young adult male is presented with preoperative roentgen diagnosis on a small bowel follow-through. Anatomic types of paraduodenal fossae are described and some points of surgical importance are stressed.


Venous intravasation of pantopaque during myelography. Report of two cases and a review of the literature.

Pantopaque venous intravasation, an uncommon complication of myelogram, has been reported since 1945. Traumatic spinal puncture has been known to be associated with Pantopaque venous intravasation, but its close relationship with the level of spinal puncture at the L5-S1 intervertebral disc space has not been emphasized. Spinal puncture at that level should be discouraged in order to prevent Pantopaque venous intravasation and pulmonary embolism during myelography.


Gastric leiomyoblastoma in a child.

A gastric leiomyoblastoma can occur in any age group, but the prognosis and sex distribution are different in the child-adolescent group than in adults. In the adult, the lesion is generally considered to be benign, though in the child-adolescent group it may metastasize to regional lymph nodes. The possibility of a leiomyoblastoma is suggested by multicentric intramural gastric tumors discovered in a female child or adolescent who has had gastrointestinal hemorrhage.


Spontaneous nephro-duodenal fistula.

Nephroduodenal fistulas are an uncommon clinical entity characterized by upper abdominal pain and associated with pyelonephritis and renal calculus formation. Most of the recent cases reported are secondary to inflammatory changes in the kidney. Two cases are reported here, one of a nephroduodenal fistula and another of a combined nephroduodenal and nephrocolic fistulae. Carefully monitored X-ray studies determined in both cases the exact site and extent of the fistulae. The optimal treatment is nephrectomy.


Resident evaluation - is our method effective?

In an earlier paper, "Resident Evaluation--Is Your Method Effective?", it was emphasized that the objectives for the residency program in Radiology should be clearly delineated and used to judge the performance of the resident in all stages of the training program. After having had approximately three years' experience in the use of the procedures outlined in the original paper, we are recommending some changes and modifications. The purpose of the evaluation is to provide clear and detailed evidence of the performance of the resident over the period of his training program. As outlined in the previous paper, the basic principles -- learning is a very individual matter, learning depends on the motivation of the individual, and learning depends on the satisfaction in the learning task--remain unaltered. Changes have been made in the design, execution and details of evaluation. The final sucess of any training program will necessarily be measured by the degree to which the residents attain the outlined objectives of the program and the degree to which the graduates of the program are accepted into the specialty by the accrediting bodies.


Half-body radiotherapy of advanced cancer.

Half-body radiotherapy to deliver doses up to 1000 rads in a single exposure evolved to meet a need in treating patients with advanced cancer. Large fields are required and a high radiation output. Treatments are well tolerated, although radiation sickness occurs in 80% of upper half and 33% of lower half-body treatments. Marrow depression is not a problem and the second half body can be treated when the peripheral blood count has returned to normal. Subjective improvement occurs in most patients and this group had a median survival of six months.


Localization of gallium-67 citrate in salivary glands following radiation therapy.

Gallium-67 citrate was found to localize in the salivary glands of eight patients after therapeutic doses of radiation to a treatment field including the glands. In two patients, autopsy evidence to the effect that no primary disease was present to explain such uptake was obtained. In another two patients the abnormal uptake was not observed on follow-up. The recognition that radiation sialadenitis is a potential cause of false-positive gallium scintiscans is important in their interpretation.


Ataxia-telangiectasia.

The clinical and radiographic features of twenty-two cases of ataxia-telangiectasia are reviewed. Severity of pulmonary manifestations tended to correlate closely with severity of immunodeficiency. Observations are made upon the problem of intercurrent malignancy in ataxia-telangiectasia. The radiographic studies of patients seen at the National Institutes of Health with A-T were reviewed retrospectively and correlated with degrees of immunodeficiency determined clinically. Three fairly distinct groups with high, intermediate and low incidence of sinopulmonary disease were found. Comment is made on the problem of increased malignancy in A-T and possible clues in the detection by the radiologist.


Angiographic demonstration of a thoracic aortic aneurysm with rupture into the pulmonary artery.

A case of rupture of a thoracic aortic aneurysm into the left pulmonary artery is presented. This is an unusual complication of an aortic aneurysm. The importance of angiography in its accurate and early diagnosis is described.


Roentgen features of the rheumatoid mid- and hindfoot.

The roentgen features of rheumatoid involvement of the mid- and hindfoot are analyzed utilizing anatomic and pathologic material from "control" and rheumatoid cadavers and a detailed radiographic study of a rheumatoid population. Characteristic findings include diffuse abnormalities predilecting the calcaneus, talocalcaneonavicular and tarsometatarsal joints with joint space narrowing, bony fusion and reactive sclerosis. Rheumatoid localization in the mid- and hindfoot is not unusual, particularly in long-standing disease, and is almost universally associated with metatarsophalangeal alterations.


Lipid vesicle-cell interactions. I. Hemagglutination and hemolysis.

The interaction of lipid vesicles (liposomes) of several different compositions with erythrocytes has been investigated. Lecithin liposomes, rendered positively charged with stearylamine, exhibit potent hemagglutination activity in media containing low concentrations of electrolytes. The hemagglutination titer is found to be a linear function of the zeta potential of the lipid vesicles. Hemagglutination is reduced when the surface potential of the cells is made more positive by pH adjustment or enzyme treatment. Similarly, hemagglutination is reduced by increasing concentrations of electrolytes. Hemagglutination is examined theoretically and is shown to be consistent with vesicle-cell interactions that are due to only electrostatic forces. Vesicles containing lysolecithin in addition to lecithin and stearylamine cause lysis of erythrocytes, provided the lipids of the vesicles are above the crystal-liquid crystal phase transition temperature. In addition, hemolysis requires close juxtaposition of the vesicle to the cell membrane; vesicles precoated with antibodies exhibit severely diminished hemolytic activities, only a small fraction of which can be attributed to a reduction in hemagglutination titer. Evidence is presented indicating that a single vesicle is sufficient to lyse one cell. With regard to hemagglutination and hemolysis, lipid vesicles of simple composition mimic paramyxoviruses such as Sendai virus.


Ultrastructure of mitosis in the cowpea rust fungus Uromyces phaseoli var. Vignae.

Aspects of the ultrastructure of mitotic nuclei of the fungus Uromyces phaseoli var. vignae are described from both intercellular hyphae in the cowpea host and infection structures induced to differentiate in vitro. The interphase nucleus-associated organelle (NAO) consists of two trilamellar acircular disks connceted by an osmiophilic bar. The intranuclear spindle develops between these disks when they separate. The spindle contains pole to pole, interdigitating, chromosomal, and fragmentary microtubules arranged to form a central bundle along the surface of which lie the metaphase chromosomes. No metaphase plate is found. There are up to three microtubules per kinetochore and approximately 14 chromosomes on the haploid spindle. Telophase elongation appears to involve extension of pole to pole microtubules with no evidence for the remaining presence of interdigitating microtubules. Concomitantly, numerous cytoplasmic microtubules develop from each NAO disk where few or none are present in other phases. Reformation of the interphase NAO involves the formation of a sausage-shaped intermediate at late telophase. The nuclear envelope remains intact and the nucleolus persists throughtout division. Various aspects of the spindle and NAOs appear to be evolutionary intermediates between Ascomycetes and higher Basidiomycetes, thus supporting the theory of Basidiomycete evolution from the former group and demonstrating an encouraging correlation between mitotic characteristics and other phylogenetic markers.


Presence of adenylate cyclase activity in Golgi and other fractions from rat liver. II. Cytochemical localization within Golgi and ER membranes.

The presence of adenylate cyclase (AC) in liver Golgi and microsomal fractions from ethanol-treated rats was tested cytochemically using 5'-adenylyl imidodiphosphate (AMP-PNP) lead phosphate method. Parallel biochemical assays showed that rat liver Golgi AC was only partially inhibited by lead: in the presence of 1 mM Pb++ 80% of the enzyme was preserved, while when 2 mM Pb++ was used 25% remained. No cAMP was formed when the AMP-PNP medium was incubated in the presence of 1 or 2 mM Pb++ but in the absence of cell fractions, indicating that at these concentrations Pb++ does not cause the nonenzymatic hydrolysis of AMP-PNP. Therefore, the reaction product observed by cytochemical localization is not due to the nonenzymatic hydrolysis of AMP-PNP by Pb++. In Golgi subfractions, lead phosphate reaction product was widely distributed among Golgi elements: it was seen in association with the majority of the very low density lipoprotein-filled secretory droplets which predominated in the two lightest Golgi fractions (GF1 and GF2) as well as within the majority of the cisternae found in the heaviest Golgi fraction (GF3). In the latter, reaction product was heaviest along the dilated peripheral rims of the cisternae. In all cases, the reaction product was localized to the outside or cytoplasmic face of the Golgi membranes. When microsomes were incubated cytochemically for AC, deposits were found on the cytoplasmic surface of smooth endoplasmic reticulum (ER) membranes, but none were observed on rough ER membranes. The results confirm the biochemical data reported previously indicating the presence of AC in Golgi and smooth microsomal fractions from rat liver and further demonstrate that the activity is indeed indigenous to Golgi elements and not due to plasma membrane contaminants. They also indicate that AC is widely distributed among Golgi and smooth ER elements. Thus, AC is not restricted in its distribution to plasma membranes as usually assumed.


Exocytosis in secretory cells of rat lacrimal gland. Peroxidase release from lobules and isolated cells upon cholinergic stimulation.

Release of peroxidase from secretory cells of rat lacrimal gland upon cholinergic stimulation was studied in vitro with single lobules and isolated cells (lacrimocytes). Isolated lobules, kept in Eagle's medium, remain structurally intact and reaction product of peroxidase is confined to cisternae of rough endoplasmic reticulum, elements of the Golgi apparatus, and all secretory granules. Morphologically, exocytosis occurs by membrane fusion and discharge of granule content. The highest rate of peroxidase released from lobules is observed at 10(-4) M carbamylcholine. The specific activity of peroxidase released into the medium is fourfold higher as compared to the lobules. Release of peroxidase is suppressed by atropine when added before or after the addition of carbamylcholine. At 4 degrees C, no peroxidase release occurs upon cholinergic stimulation. The exocytotic release of peroxidase is dependent on energy supply, as indicated by substantial inhibition (at 37 degrees C) under anoxic conditions or in the presence of dinitrophenol, KCN, or carboxyatractyloside. Furthermore, the process is sensitive to colchicine and vinblastine. Isolated lacrimocytes, consiting of 95% secretory acinar cells, are prepared by digestion with collagenase, hyaluronidase, and trypsin. They retain the characteristic polarity of secretory cells in situ, and localization of peroxidase is the same as in lobules. Since isolated lacrimocytes respond to cholinergic stimulation in the same way as lobules, the receptors are not damaged by the isolation procedure and appear to be associated directly with the exocrine cell. Oxygen uptake by isolated lacrimocytes is about 14 nmol O2 X min-1 X 10(-6) cells; it is about doubled by uncoupling with dinitrophenol. Oxygen uptake rises by 20-30% above the resting rate upon cholinergic stimulation. This additional uptake is suppressed by atropine or by added cholinesterase, indicating that continuous receptor occupancy may be required for the energy demand by exocytosis. On the basis of the specific activity of peroxidase in the medium, the energy demand resulting from cholinergic stimulation is estimated to be 0.08 mumol ATP (or energy-rich phosphate bonds) per microgram of protein released from the lacrimocytes.


Initial attachment of baby hamster kidney cells to an epoxy substratum. Ultrastructural analysis.

In the presence of serum-containing medium, BHK cells attached and spread during a 1-h period onto a 3-5 nm thick serum layer absorbed on the substratum surface. The closest approach of the plasma membrane to the serum layer was observed to be about 9nm, which was determined by tilting the sectioned cells in a goniometer holder. Bundles of microfilaments or other cytoplasmic specializations were not observed in association with the regions of close contact. However, in the space between the plasma membrane and the adsorbed serum layer, a diffusely stained material could be visualized after fixation/staining by the tannic acid-glutaraldehyde technique. This technique also permitted increased clarity of visualization of trilaminar appearance of the plasma membrane. The distribution and mobility of anionic sites on the surfaces of attached and spreading cells was determined by labeling with polycationic ferritin. We observed movement of polycationic ferritin into large clusters on the cell surface, collapse of cell surface microextensions, and endocytosis, all of which were similar to our previous findings utilizing cells in suspension. However, the absolute amount of ferritin bound to the upper cell surface was less than that previously observed when suspended cells were put under similar labeling conditions. Also, polycationic ferritin did not appear to penetrate between the lower cell surface and the substratum.


Transport and countertransport of thymidine in ATP depleted and thymidine kinase-deficient Novikoff rat hepatoma and mouse L cells: evidence of a high Km facilitated diffusion system with wide nucleoside specificity.

Incubation of cultured Novikoff rat hepatoma and mouse L cells in a glucose-free basal medium containing 5 mM KCN and 5 mM iodoacetate for about 10 minutes resulted in a complete depletion of the cells of ATP. ATP-depleted wild type cells or thymidine kinase-deficient sublines of Novikoff or L cells took up thymidine rapidly from the medium without concentrating it intracellularly, and exhibited countertransport of thymidine. Thus uptake was by facilitated diffusion. This transport system differs from the substrate-specific, low-Km (0.5 muM] thymidine transport system previously described for various types of cultured cells in that it exhibits an at least 100-fold higher Km and transports equally well various ribo- and deoxyribonucleosides. The results suggest that the rate-limiting step in thymidine incorporation into the nucleotide pool by wild type cells is phosphorylation rather than transport, or that the cells possess two transport systems, a facilitated diffusion system with low substrate specificity and a second system which involves substrate phosphorylation by thymidine kinase.


Density regulation of amino acid transport in cultured, androgen-responsive tumor cells.

In androgen-responsive cells, activity of a sodium-dependent transport system for neutral amino acids is shown to be density-dependent, whereas activity of a sodium-independent system is not.  Transport of the non-metabolizable amino acids, alpha-aminoisobutyric acid )alpha-aib) was largely sodium-dependent, wherease transport of cycloleucine (cyleu) was by  both sodium-dependent and sodium-independent systems.  The km for cyleu transport changed with density because only the sodium-dependent portion of this transport was density-dependent.  By contrast, the Vmax for alpha-aib transport changed with density, indicating that either amount of functional membrane carrier of the activity of the sodium pump was affected.


Density regulation of amino acid transport in cultured, androgen-responsive tumour cells.

In androgen-responsive cells, activity of a sodium-dependent transport system for neutral amino acids is shown to be density-dependent, whereas activity of a sodium-independent system is not. Transport of the non-metabolizable amino acid, alpha-aminoisobutyric acid (alpha-aib) was largely sodium-dependent, whereas transport of cycloleucine (cyleu) was by both sodium-dependent and sodium-independent systems. The Km for cyleu transport changed with density because only the sodium-dependent portion of this transport was density-dependent. By contrast, the Vmax for alpha-aib transport changed with density, indicating that either the amount of functional membrane carrier or the activity of the sodium pump was affected.


Density-dependent regulation of proliferation rate in cultured, androgen-responsive, tumour cells.

Short-term cultures of androgen-responsive Shionogi 115 (S115) cells exhibited density-dependent regulation of proliferation rate in the presence or absence of testosterone. The average surface area per cell exposed to the growth medium was inversely proportional to population density. By contrast, long-term cultures (serially passaged in testosterone-containing medium for several months) did not exhibit density-dependent regulation of proliferation rate when grown in testosterone-containing medium. In this medium, cells became elongated and no longer exhibited any obvious decrease in exposed surface area with increasing density. Nevertheless, when subcultured into testosterone-free medium, these cells reverted to an epithelial morphology and exhibited density-dependent regulation of proliferation rate. These relationships suggested that the proliferation rate of cells decreased with density in proportion to the decrease in exposed surface area...


Cell cycle dependent changes in potassium transport.

K transport has been investigated during progression of cultured Ehrlich ascites tumor cells through the cell cycle. Using a double thymidine block technique, Ehrlich cells carried in continuous culture have been synchronized, as verified by simultaneous monitoring of cell number, cell volume, 3H-thymidine incorporation and mitotic index. Unidirectional influx, efflux and cell content of K have been monitored throughout the cell cycle. The nature of the pump mediated, ouabain-sensitive K flux and the furosemide-sensitive component of K flux, presumably representing K-K exchange, have also been evaluated. In early S period the ouabain sensitive component, representing the Na-K pump, comprises 52% of the total unidirectional K influx and subsequently declines during G2 period to a minimum of 40% in mid G2. During M and early S the activity again rises. As the ouabain sensitive component becomes maximal in late S period, the furosemide sensitive component diminishes from approximately 30% of the total influx to approximately 10%. The same pattern is observed in the G2 period. As the pump component diminishes, the furosemide sensitive component increases. Furosemide sensitive K efflux has also been monitored and the pattern is equivalent to that observed in the invlux studies. No change in net K flux is observed in the presence of furosemide. This indicates that the furosemide sensitive component represents an exchange component for K. These results are consistent with the conclusion that the alterations in exchange and pump fluxes are physiological events associated with progression of the cell cycle.


Reversible independent alterations in glucose transport and metabolism in cultured human cells deprived of glucose.

We have measured uptake of 3H-hexoses into diploid human cells by exposing them to brief pulses of isotopic sugar during the log-growth, subconfluent-growth, and confluent-growth (contact inhibited) phases of the strain HSWP derived from human skin. 3H-deoxyglucose appears to be taken up three times faster than 3H-glucose. After exposure to 3H-glucose for longer than one minute, the cells excrete approximately 70% of the isotope into the medium as lactate. If lactate production (and hence excretion) is abolished by treating the cellls with iodoacetic acid or dinitrofluorobenzene, neither of which inhibits transport, the uptake of 3H-glucose is found to be in fact somewhat larger than that of 3H-deoxyglucose. If cells are deprived of glucose for 24 hours, apparent uptake of 3H-glucose is enhanced 10-fold or more. This latter increase is accounted for by 2- to 3-fold enhancement of true transport plus retention of greater than 90% of the radioactivity, since little lactate is formed or excreted in glucose-deprived cells. Deoxyglucose, galactose, or pyruvate when present during glucose deprivation each have quantitatively different effects on the cells' capacity to produce lactate from a short pulse of glucose, but none of them prevents the enhancement of hexose transport. After restoration of 5 mM glucose to starved cells, their metabolsim returns to normal (in the sense that approximately 70% of the glucose taken up in a pulse is again excreted as lactate), with a half-time of 0.5 hour; but the transport of hexoses returns to control levels much more slowly, with a half-time of approximately 6 hours. The two processes appear to be independently regulated.


Effect of ESF preparations on different types of erythroblasts.

A new method of quantitative (14)C-autoradiography was applied for evaluating possible effects of erythropoietin(ESF) on the DNA synthesis rate of differentiated erythroid murine bone marrow cells identified as proerythroblasts, basophilic and polychromatic erythroblasts...


Effect of 5-bromodeoxyuridine on the appearance of cytoplasmic poly-A containing RNA.

Cytoplasmic poly-A containing RNA, synthesized by cultured chick embryo cells, was examined during growth in 5-bromodeoxyuridine. The kinetics of 3H-adenosine incorporation into this species of RNA, when compared to the rest of the cytoplasmic RNA, and to control cells, indicates that the rate of synthesis of this RNA is slower in BrdU treated cells. An examination of the rate at which a steady state distribution of radioactivity, between the poly-A segment and non-poly-A portion of poly-A containing RNA is reached also indicated that this species is synthesized at a lower rate in BrdU treated cells.


Growth abnormalities of cultured human skin fibroblasts derived from individuals with hereditary adenomatosis of the colon and rectum.

A heritable propensity to develop malignant lesions is found in individuals with familial adenomatosis of the colon an rectum (ACR) and the Gardner's syndrome variant, an autosomal dominant trait. In the present study, the growth characteristics of cultured skin fibroblasts (SF) derived from normal-appearing flat skin biopsies of ACR families, representing all phenotypes, and appropriate controls were investigated. SF were obtained from stocks between the second and fifth passages and growth to confluency in Eagle's Minimal Essential Medium (EMEM) supplemented with 15% fetal calf serum (FCS). Following trypsinization, cells were replanted in EMEM supplemented with either 1% or 15% FCS at an initial density of 4 x 10(3) cells/cm2 and counted daily for five days. Normal SF representing several age groups (both sexes) and those obtained from non-afflicted individuals of ACR families grew only in 15% FCS. In contrast, SF from ACR subjects and from embryonal skin grew both in 1% and 15% FCS. SF from several clinically asymptomatic adults, children or ACR patients, grew in 1% FCS as well. Cell cultures from ACR individuals showed regions of criss-crossed arrays and multilayered pattern. These growth properties were not observed in normal cell cultures. The SF from ACR individuals did not grow in methocel, nor did they form tumors in athymic mice. These results suggest the occurrence of previously undetected biochemical alterations in SF taken from ACR genotypes.


Ornithine decarboxylase inactivation in HeLa cells.

Ornithine decarboxylase (ODC) can be induced up to 100-fold over basal levels four hours after addition of glutamine to the medium of HeLa cells growing in suspension culture. As demonstrated in several other cell types, ODC is inactivated very rapidly in HeLa cells, and the rate of inactivation is seen to vary with a half life of 9-15 minutes in uninduced cells and rises to ca. 60 minutes at the peak of induction. Quantitatively, the change in rate of inactivation cannot completely account for the observed rise in activity, thus synthesis or activation of ODC must also be involved in the induction process. The inactivation process requires metabolic energy and it can be sustained by glycolytic derived energy. Other factors which are known to inhibit protein breakdown in mammalian cells, such as sodium fluoride, insulin, or tosyl phenylalanyl chloromethyl ketone, had no effect on the rate of inactivation of ODC. Attempts to demonstrate ODC inactivation in a cell free system at neutral pH were unsuccessful.


Selective isolation of reversible cold sensitive variants from Chinese hamster ovary cell cultures.

Variants of the Chinese hamster ovary cell line CHO-KI (ATTCCCL61)which grow almost normally at 38.5C but very poorly or not at all at 30C were obtained after treatment with mutagens and application of an indirect selection procedure.  Two kinds of variants were recovered.  In the first of these, the cold sensitive phenotypw is expressed completely only at low cell densities.  At higher cell desity, growth continues at the nonpermissive temperature, but at a reduced rate.  In the second class, the cold snesitive phenotype is independent of cell density.  Two members of the  latter class were studied in detail.  In both lines, after shift to the nonpermissive temperature, the rateof H-thymidine incorporation declines markedly; the rates of H-uridine and H-phenylalanine uptake are less drastically reduced.  Autoradiographs indicate that the decline in thymidine uptake at the nonpermissive temperature is due to an elongation of part of the cell cycle, so that a smaller proportion of the cells lie in the synthetic (S) phase of the cell cycle with a consequent reduction in the fraction labeled cells.  The uridine labeling patterns of the mutants appear to rule out a ribsomal lesion.  Low temperature growth inhibition of both cell strains was reversible.  In one of the cell lines, an apparent stretching of the cells at the low temperature produces substantial alterations in cell shape.


Growth of in vitro colony-forming cells from normal human peripheral blood leukocytes cultured in diffusion chambers.

The growth of granulopoietic progenitor cells (CFU-C) in diffusion chambers during culture of peripheral blood leukocytes from 10 normal subjects has been studied. At various times after initiation of diffusion chamber culture, cells harvested from the chambers were transferred to agar culture for measurement of CFU-C concentration. Under these conditions colonies could be grown successfully in agar culture provided pronase, necessary for the chamber harvesting procedure, was first removed by careful washing. A marked increase in the number of CFU-C, up to 25-fold the initial value, was observed in 8 out of 10 subjects. Here the growth pattern was similar, independent of the initial CFU-C throughout the diffusion chamber culture period was very poor. The growth of CFU-C from a given individual's blood was shown to be reproducible in repeated studies in 2 subjects, one of whom showed a proliferative and the other a non-proliferative pattern. Evidence suggests that the increase in CFU-C in diffusion chambers is the result of both self-renewal of these cells and influx from a more primitive compartment, although the present data do not allow an estimate of the relative magnitude of each.


[Wirsung's duct and chronic pancreatitis. Changes in the morphology of the pancreatic ducts after pancreatic duct shunts. 23 cases followed-up].

The authors study in chronic pancreatitis the morphology of Wirsung's duct in 31 patients who had undergone repeated operations, 23 of them were submitted in a first stage to an anastomosis between the pancreatic duct and the digestive tract. The main causes of failure were obstructions of the anastomosis, biliary complications and continuation of the pancreatic disease. The difference in prognosis between pancreatitis with a dilated pancreatic duct, and those with a filiform duct, is perhaps due to lesions of different histological appearance and course. The best results were obtained in patients able to give up alcohol and in whom it was possible to carry out a broader anastomosis on a dilated and unobstructed pancreatic duct.


[Surgical treatment of stage II arteriopathies of the lower limbs].

The authors report their results of 218, operations carried out in patients at stage II of peripheral arterial disease during the period 1970 to 1974. In 138 patients who were submitted to direct surgery, the results appeared specially satisfactory in cases of aorto-iliac and ilio-femoral obstruction (83 and 87 p. 100 good results at 4 years). They were still 76 p. 100 in cases of femoro-popliteal obstruction.


[Contribution of scintigraphy in acute and chronic pancreatitis. Apropos of 200 examinations].

The authors report 200 examinations and discuss the role of radio-isotope scans of the pancreas with electronic subtraction of the liver image in acute and chronic pancreatitis. As an emergency, the examination is of definite diagnostic value, the absence of fixation with a suggestive clinical context, is strong evidence of pancreatic disease and normal uptake eliminates the latter. On the other hand, in painful syndromes in which chronic pancreatitis is suggested, the diagnostic interest is very limited for the low uptake on radio-isotope scan occurs at a late stage and is not specific. Radio-isotope scans are worth a place in exploration of the pancreas in acute pancreatitis.


[Revascularization of arteries of the leg by bypass in chronic arteriopathies of the lower limbs. Apropos of 50 cases].

The authors report 50 by-pass operations followed up for from 6 months to 4 years, which were designed to restore the blood supply via the leg arteries in peripheral arterial disease of the lower limbs. 40 patients were at stages III and IV; acute ischemia required emergency operation in 6 cases. 2 Patients died immediately after operation. 30 by-passes remained permeable. A cure of the trophic disorder and a functional improvement were obtained in 39 patients with only 8 subsequent amputations. In the light of the world literature and their personal experience they discuss the main pronostic factors and the indications for this method which has proved to be a remarkable progress in the treatment of severe arterial disease of the lower limbs.


[Amputations for arteritis. Criteria of choice of the level of the amputation].

The practice in arteritis, of economic amputations conserving the knee, is justified by the better quality of the functional results obtained and, also, the risk of later bilateral amputation. This risk, in the series studied here was estimated at 21.6 p. 100. This conservative attitude does, however, have certain risks, the main risk being the necessity for a new amputation at a higher level. The proportion of failures leading to high mid-thigh amputation was of the order of 22 p. 100. To ensure the best possible security in economic amputations, it is important to respect a certain number of criteria which are studied here: the general and psychological condition of the patient, the arteriographic findings, the functional investigations and, finally, the operative test which assesses the vitality of the tissues at the level of the area of section.


[Etiopathogenesis of varicocele and therapeutic conclusions].

The many pathogenic theories for varicocele, mostly hypothetic, have shown the uncertainty of the cause of the disease including the use of the word "essential". Anatomical, phlebomanometric and phlebographic studies determined the main cause of varicocele, the short course of the aorto-mesenteric artery which, by stricture of the left renal vein in the standing position, produces reno-spermatic venous reflux producing varicose dilatation of the left spermatic plexus and, thus, varicocele. In addition to this main cause, the vascular background with a qualitative deficiency of the connective tissue and prolonged standing during work, are the main factors which favour the onset of varicocele. An active, early therapeutic attitude and high ligature of the left spermatic vein give the best results.


[Jejuno and ileo-ileal invaginations revealing mutiple melanic metastases of the small intestine. Case report].

The authors report the case of 60 year old man with multiple jeuno-ileal metastases from a malignant melanoma of the skin revealed by jejuno-jejunal intussusceptions. In the light of this case, the authors emphasize the frequency and preferential localisation of these metastases at the level of the jejunum and ileum, the usual mildness of their symptoms until the day where acute obstruction requires laparotomy. The multiple lesions, their diversity and their very rapid course.


[Treatment of cutaneous nevocarcinomas].

For long, malignant melanoma of the skin was never treated other than surgically; in spite of considerable effort in this field, a large number of them remained with a very poor prognosis. In the severest cases, it was noted that the prognosis was little influenced by recent therapeutic progress. The latter, however, in the light of recent experience, seemed to justify the hope which was placed in it initially. The object of this report is to relate modern progress in the treatment based on clinical and pathological findings. It is the result of work undertaken during recent years by the research group on malignant melanomas of the Lariboisiére-Saint-Louis hospitals.


[Research on intrazygoportal disconnection by use of Vosschulte's ring. Experimental study in the dog].

The disappointing results of porta-caval anastomosis in the treatment of portal hypertension have led surgeons to treat directly oesphageal varices by interazygoportal disconnection through Vosschulte's ring. An experimental study on 29 dogs, permitted the authors to make several discoveries. The ring introduced through the pylorotomy should have a caliber identical to that of the oesphagus. The course of the stricture then occurs towards mutilant annular fibrosis of all the oesphageal wall. The marked morbidity due to regurgitation linked to stenosis, may be reduced by early instrumental dilatation.


[Pre-operative irradiation in the treatment of cancers of thoracic esophagus].

The authors analyse an homogeneous series (2 operators) of 650 patients operated on for thoracic oesophageal carcinoma, of which 185 were submitted to pre-operative irradiation with cobalt. Irradiation led to a definite increase in the resectability rate 74 p. 100 as against 62 p. 100. There was an increase in the mid-term survival rate, especially an increase of 5 p. 100 at 2 years, 8 p. 100 at 3 years, 6 p. 100 at 4 years; The 5 survival rate was little modified. It was 12 p. 100 for 290 cases, resection alone, and 15 p. 100 for 137 resections after irradiation.


[Surgical treatment of villous tumors of the rectum].

Out of 36 villous tumours observed in 31 patients, 50 p. 100 were sessile and 1/3rd were degenerated, their degeneration was much more frequent in the case of sessile villous tumours. There were 5 biopsy errors. Out of 23 villous tumours, class A, 11 underwent local removal with two relapses, 7 recto-sigmoidal resections and 3 amputations of the rectum and 2 Hartmann operations. Out of 12 villous tumours classified as B and C, 5 underwent local removal, 4 resections, 3 palliative amputations. In all, there were 5 relapses. Tumours class C are severe and have the same prognosis as carcinoma of the rectum. Certain benign villous tumours are very difficult to remove locally owing to their broad base and their volume, which may necessitate mutilation. The present therapeutic attitude of the authors is due to the severity of relapses which may be seen late and possible biopsy errors. Broad removal is recommended for benign villous tumours which are broadly implanted and very voluminous. If the villous tumour is above the pouch of Douglas, resection is preferable to local removal. On the other hand, local removal is justified for benign villous tumours placed above the pouch of Douglas, which may be pediculated or sessile with a narrow base and with normal neighbouring mucosa.


[Uretero-ileoplasty in bilharzian uretero-hydronephrosis].

The authors report 22 cases of uretero-ileo-plasty for uretero-hydronephrosis due to schistosomiasis. The indications for operation depend on the following criteria: the degree of dilatation which varies from simple atonia to very large hydronephrosis which one must not wait for, ureteral stenosis, vesico-ureteral reflux, the degree of renal failure assessed by studies of creatinine and urea clearance and the resistance to treatment. The operative technique which is not specific for bilharziasis includes uretero-ileo-plasty which is often bilateral, for even in asymmetrical cases, the least affected ureter is often of poor quality. There were failures in two cases due to irreversible renal failure, and in two cases, due to peritonitis. The late results of the other cases appear very favourable: increased vesical capacity, diminution of cystalgia, comfort and improved, general health, were the main factors. Stenosis of the anastomosis, vesico-ureteral reflux and urinary infection, acidosis, lithiasis are rare or not very severe.


[Diagnostic traps and procedure to follow in radio-invisible biliary calculi].

Instead of the term biliary microlithiasis, the authors prefer the general concept of radio-invisible biliary lithiasis. The size, the homogeneity, the site and chemical composition of the gall stones, which are the usual criteria of biliary microlithiasis, also are valid for radio-invisibility; thus the diagnostic traps are the only original characteristics of a pathology with hazy outlines, defined as one which escapes well conducted a radiological exploration. Biliary micro-lithiasis therefore raises a triple problem. The indications for biliary surgical exploration, in spite of normal cholangiography, seem to us undoubted in acute relapsing pancreatitis, once the usual medical causes have been eliminated. It should be more relative in pure biliary pain which first requires, not only repeated cholangiography, but also constant clinical and laboratory signs which suggest that the symptoms are organic and that gall stones are in formation. After laparotomy, cholecystectomy is required, not only in perceptible lithiasis, but also when the gall bladder bile contains mud, débris, sand or cholesterol spheroids. In the other cases, the choice between abstention or cholecystectomy depends on the clinical and laboratory context. The surgical attitude with regard to the common bile duct is discussed in three possibilities depending on the case, e.g. routine exploration, abstention or exploration in certain cases.


[Synovectomies of the knee. Apropos of 47 cases tuberculous synovitis].

The authors carried out synovectomy of the knee in 70 cases, including 47 for tuberculous synovitis, with follow up periods of 2 years and half to 18 years. The subjects included: -30 children from 2 to 15 years; -17 adults from 16 to 50 years, with 3 cases of more than 50 years. Very good: greater or equal to 110 degrees in 2 cases; -Good: greater or equal to 90 degrees in 26 cases; -Average: greater or equal to 45 to 90 degrees in 11 cases; -Poor: less than 45 degrees in 8 cases. The authors believe that synovectomy in tuberculous synovitis of the knee without an osteocartilagenous lesion, or with slight lesions of the joint is more efficacious than medical treatment alone and may shorten the duration of treatment.


[Stenosis of the small intestine after strangulated hernia].

Return to the abdominal cavity of an intestinal loop which had suffered from ischemia, may induce, after a free interval, disordered transit or frank intestinal obstruction, sometimes peritonitis. This is always dangerous, especially in elderly patients. The authors report 5 cases which illustrate this danger, in daily surgery, and recall the rules for treatment of strangulated hernia.


[5 kg sternal chondroma, treated by resection and silastic prosthesis. Satisfactory result].

Resection of the sternum and adjacent, cartilages raises difficult problems of reconstruction. The tumour, which, exceeding 5 kg was considered histologically as a chondroma, was replaced by a silastic prosthesis with an excellent result 13 months later.


[Route of approach of distal deep femoral artery].

Proximal implantation of by-passes between the femoral and popliteal or femoral and leg arteries, may be carried out on the middle portion of the profunda femoris artery, between the first and second perforating arteries. An approach some distance from Scarpa's triangle, permits one to reach fairly easily, within a muscular triangle between the vastus medialis and the adductor longus. Two cases illustrate the interest of this route of approach described in Lyons for the first time in 1970.


[Our indications in surgical treatment of supra-condylar fractures of the femur].

The treatment of supracondylar fractures of the femur should be surgical. The authors consider the requirements of a strict technic of osteosynthesis. The use for this purpose: either a plate and strip, or a plate and screw, and justify their respective indications.


Gas chromatographic analysis of heptafluorobutyryl derivatives of some carbamate insecticides.

The gas chromatography (GC) of a number of carbamate insecticides as their heptafluorobutyryl derivatives has been studied. The reaction makes use of heptafluorobutyric anhydride with trimethylamine in benzene as a catalyst. Conversion of the insecticides to their products is complete in 15-20 min at room temperature. The same reaction proved satisfactory for trifluoroacetylation. The excess reagent and trimethylamine are removed by partitioning with water in the reaction test tube. An aliquot of the organic phase is used for analysis. The heptafluorobutyryl derivatives are stable for several days on the lab bench without removal of the aqueous phase from the test tube. GC detection was carried out with a Coulson conductivity detector in the halogen (reductive) mode. About 1.0 ng produced a 50% full-scale response for most of the carbamates (3 min retention time) on 3% OV-1. This method was applied to the analysis of several carbamates spiked in corn, mustard greens, turnip greens, lettuce and cabbage.


Sephadex LH-20 multiple-column chromatography for the simultaneous separation of progesterone, deoxycorticosterone and 17alpha-hydroxyprogesterone from small plasma samples.

A simple and convenient chromatographic method is described for the simultaneous and complete separation of the unconjugated steroids progesterone, deoxycorticosterone and 17 alpha-hydroxyprogesterone on 40-cm columns of Sephadex LH-20 using a water-saturated solvent system containing n-heptane-chloroform (1:1) plus 0.25% of ethanol. Manual operation of up to ten columns run in parallel is facilitated by the use of graduated, cylindrical, solvent reservoirs on top of the columns. Thus, negligible column-to-column, and only limited day-to-day, variations occur in the elution patterns, and constant high recoveries are obtained. When combined with a previously described 60-cm LH-20 chromatography, eight important corticosteroids can be isolated individually from 16 to 20 small plasma samples per day.


[Injuries of Lisfranc's joint: severe sprains, dislocations, fractures. Study of 39 personal cases and biomechanical classification].

A study of 39 cases permitted us an analysis of the lesions of Lisfranc's joint, from severe fracture dislocation to fractures which are rarely reported in this joint. Standard XRay views seem to us insufficient and we propose carrying out oblique views and, sometimes, tomographic views. We were thus able to obtain information permitting a mechanical functional classification, and thus interpret the lesions in relation to the cause and thus adapt treatment. A long-term follow up of 23 cases suggests frequently invaliding sequelae after involvement of the tarso-metatarsal joints. The authors suggest resection and arthrodesis straightaway in dislocations.


Thin-layer chromatography for detection of peptide cleavage or integrity during reactions of the Z-alanylglycines with aniline or phenylhydrazine under papain catalysis.

Z-l-Ala-gly and Z-dl-ala-gly can yield anilides and phenylhydrazides through peptide cleavage or peptide integrity during papain-catalyzed reactions with aniline (NH2Ph) or phenylhydrazine (NH2NHPh). Since Z-d-ala-gly yielded only uncleaved Z-d-ala-gly-NHPh or Z-d-ala-gly-NHNHPh, these were used as standards in thinlayer chromatography (TLC) for detecting integrity. Known Z-l-ala-NHPh and Z-l-ala-NHNHPh were the standards for cleavage. Depending on the time of incubation, cleaved or unsplit products, or both, were readily detected. The solvent systems that were used for TLC were also effectively employed for separations of reasonable amounts of mixtures of cleaved and uncleaved products through thick-layer chromatography on Chrom AR. This was followed by isolation of the separated components.