Oxygen equilibrium curve of normal human blood and its evaluation by Adair's equation. Oxygen equilibrium curves of fresh, normal human blood have been measured by new methods which allow the control of pH, pCO2, and 2,3-diphosphoglycerate and which yield higher accuracy at the extremes of saturation than was possible previously. The curve determined by these techniques lies slightly to the right of the standard curve of Roughton et al. (Roughton, F.J.W., Deland, E.C., Kernohan, J.C., and Severinghaus, J.W. (1972) in Oxygen Affinity of Hemoglobin and Red Cell Acid Base Status (Astrup, P., and Rørth, M., eds) pp. 73-83, Academic Press, New York). The greatest difference is at low oxygen saturation, probably owing to the fact that the latter data were obtained under conditions which would lead to depletion of cellular 2,3-diphosphoglycerate. The range of p50 (oxygen pressure at half-saturation) values for four normal subjects was 28.3 mm Hg to 29.0 mm Hg. Adair's stepwise oxygenation scheme has been used to analyze the curves with the result that a1 = 0.1514 X 10(-1) (+/- 10%) mm-1; a2 = 0.9723 X 10(-3) (+/- 8%) mm-2; a3 = 0.1703 X 10(-3) (+/- 50%) mm-3; a4 = 0.1671 X 10(-5) (+/- 2%) mm-4 for the best of four data sets. Because these constants are very sensitive to changes in the shape of the oxygenation curve, this analysis is much more useful than p50 measurements in the investigation of the various allosteric effectors of the function of hemoglobin within the red cell.


Enzymatic properties of beta-D-galactoside alpha2 leads to 6 sialytransferase from bovine colostrum. The substrate specificity and kinetic properties of a pure sialyltransferase from bovine colostrum have been examined. The transferase appears to incorporate sialic acid into the sequence, NeuAcalpha2 leads to 6Galbeta1 leads to 4GlcNAc, which is commonly found in glycoproteins. It has a strict substrate specificity for CMP-NeuAc and forms only the alpha2 leads to 6 sialyl linkage with beta-D-galactosides. N-Acetyllactosamine (Galbeta1 leads to 4GlcNAc) and asialo-glycoproteins containing the N-acetyllactosaminyl linkage at the nonreducing ends of the oligosaccharides prosthetic groups are the best acceptor substrates. Isomers of N-acetyllactosamine with beta1 leads to 3 or beta1 leads to 6 glycosidic linkages are less than 1% as effective as acceptor substates as the beta1 leads to 4-linked isomer. Lactose (Galbeta1 leads to 4Glc) is also a poor acceptor, indicating the importance of the 2-acetamido group in the N-acetylglucosaminyl residues. The unnatural substrate beta-methyl-L-arabinopyrano-side, a five-carbon analog of beta-methyl-D-galactoside which contains no 6-hydroxyl, also acts as a poor acceptor of the transferase and the sialylated product has been partially characterized. Kinetic properties of the enzyme in the presence and absence of inhibitors suggest that the transferase has an equilibrium random order mechanism.


Purification of a sialyltransferase from bovine colostrum by affinity chromatography on CDP-agarose. CDP-hexanolamine agarose was used as an affinity adsorbent to purify a CMP-N-acetylneuraminate: beta-D-galactosyl-glycoprotein N-acetylneuraminyltransferase (EC 2.4.99.1) from bovine colostrum. Upon binding of the enzyme to the adsorbent, elution is achieved either nonspecifically, with 0.5 to 1.0 M sodium chloride, or specifically, with CDP. A highly purified sialyltransferase is obtained with a specific activity  440,000 times that of whole colostrum. Fractionation of the purified enzyme by gel filtration gives two species with different molecular weights but equal specific activities toward asialo-alpha1-acid glycoprotein (26.0 to 28.0 micronmol/min/mg of enzyme). The molecular weights of these two forms are about 56,000 and 43,000 as judged by sodium doedcyl sulfate-gel electrophoresis, sedimentation equilibrium, and gel filtration. The catalytic properties of both forms have been examined (Paulson, J. C., Rearick, J. I., and Hill, R. L. (1977) J. Biol. Chem. 252, 2363-2371). It is concluded that the lower molecular weight form may be a partially degraded species of the enzyme of higher molecular weight.


Preparation and properties of immobilized rubredoxin. Rubredoxin, one of the three protein components of the epoxidation/hydroxylation system of Pseudomonas oleovorans was immobilized by attachment to CNBr-activated agarose (Sepharose 4B). Since this represents the first reported example of the preparation of a water-insoluble derivative of an enzyme of this type, the electron transfer and physical properties of the conjugate were examined in order to allow comparison with those of the soluble enzyme. Immobilized rubredoxin exhibits all of the major spectral properties of the soluble enzyme above 300 nm, but some distortion in the 280 nm abosrbance band was observed. The immobilized enzyme accepts electrons from dithionite or form NADPH in the presence of spinach ferredoxin-NADP reductase, and upon reduction the visible absorbance is bleached. Immobilized rubredoxin mediates the reduction of cytochrome c in the presence of NADPH and spinach reductase, although it is less efficient in this role than soluble rubredoxin. The oxidation-reduction potential of immobilized rubredoxin was determined and found to be similar to that of the soluble enzyme. In the presence of 2.5 m guanidine HCL, the immobilized enzyme is considerably more stable than soluble rubredoxin toward denaturation. After anaerobic reduction, iron was readily removed from immobilized rubredoxin by washing in 0.5 m Tris base, PH 9.5 containing 0.07 M mercaptoethanol, and the resulting immobilized apoenzyme could then be reconstituted to give back a conjugate with the original iron content, as judged from its absorbance at 497 NM. Reptition of the entire reduction-dissociation-reconstitution cycle gave the same results as were obtained after the initial reconstitution.


Action of intrinsic sialidase of rat brain synaptic membranes on membrane sialolipid and sialoprotein components in situ. The sialo compounds in the synaptosomal membranes of young rat brain were specifically labeled in vivo by the intracranial injection of radioactive N-acetylmannosamine. More than 95% of the incorporated label was found in glycosidically bound sialic acid. Specific activities of sialic acid in the synaptic membrane gangliosides G71 (monosialo), GD1a (disialo), and GT1 (trisialo) were similar; labeling in GD1b (disialo) was consistently somewhat higher. The highest specific activity of rat brain sialidase was evenly distributed between "small myelin fragment" and synaptosomal membrane fractions, and ouabain-sensitive (Na+, K+)-ATPase also was concentrated in the latter fraction. The greatest amount of bound sialic acid was found in these subcellular fractions having the highest sialidase activity. A microsomal fraction was discovered to contain a small amount of bound sialic acid with a very high degree of radioactive labeling, but no sialidase. Release of sialic acid from the relatively intact membrane preparations by intrinsic membrane-bound sialidase occurred in two recognizable stages. There was a rapid initial release, complete within 30 min, of approximately equal amounts of lipid- and protein-bound sialic acid, corresponding to roughly half of the enzymatically releasable protein-boudn, and somewhat less than one-third of the lipid-bound, sialic acid. The remainder of the membrane sialidase-susceptible sialic acid was released in a second, slower stage. The intrinsic sialidase released 16 +/- 1% of the total sialoprotein and 31 +/- 1% of the total sialolipid sialic acid. Approximately the same amount of sialic acid is releasable from membrane sialolipid by the action of exogenous Vibrio sialidase; almost twice as much is releasable from sialoglycoprotein by this enzyme as compared with the intrinsic membrane sialidase. Each of the various membrane gangliosides appeared to be equally available to the action of the membrane sialidase. The results of this study indicate that both glycolipid- and glycoprotein-bound sialic acid in the synaptic membrane are releasable in situ by the action of the intrinsic synaptic membrane sialidase, and they suggest that this enzyme may act to modulate the physical properties of the membrane. In addition to influencing the rate of hydrolysis of endogenous membrane sialo compounds by intrinsic sialidase, pH had an effect on availability of protein-bound sialic acid. At acid pH, lipid- and protein-bound sialic acid were similarly available, but near neutral pH, gangliosides appeared to be attacked preferentially.


Biosynthesis of lipid-linked oligosaccharides in cotton fibers. Stimulation by lipids from pig liver. An acidic lipid fraction isolated from pig liver (Forsee, W. T. & Elbein, A.D. (1976) Proc. Natl. Acad. Sci. U.S.A. 73, 2574-2578) stimulated the incorporation of mannose from GDP-[14C]mannose into lipid-linked oligosaccharides using a particulate enzyme fraction from maturing cotton bolls. This lipid fraction did not stimulate mannose incorporation into the mannosylphosphorylpolyprenol. The 14C-oligosaccharides, formed in the presence or absence of the pig liver "acceptor" lipid, were released from the lipid-linked oligosaccharides by mild acid hydrolysis and were isolated by paper chromatography. Both sets of 14C-oligosaccharides had similar mobilities suggesting that they were qualitatively similar, except that those formed in the presence of "acceptor" lipid had much more radioactivity. The individual oligosaccharides were purified on a calibrated column of Sephadex G-25 and were then subjected to various treatments to obtain information about their structures. The molecular weights of the larger oligosaccharides ranged from about L1210 to 1720 indicating they probably contained from 6 to 10 sugar residues. Strong acid hydrolysis of the oligosaccharide with a molecular weight of 1350, followed by reduction with NaB3H4, gave a ratio of [3H]hexitol to [3H]hexosaminitol that was compatible with the molecular weight (about 5:2). However, the hexitol fraction contained small amounts of [3H]glucitol in addition to [3H]mannitol. Thus, this oligosaccharide may contain small amounts of glucose in addition to mannose and GlcNAc. When the smaller oligosaccharides (i.e. those having 5 or 6 sugars) were treated with alpha-mannosidase, essentially of all the 14C was released as mannose, but only 30 to 50% of the radioactivity could be released from the larger oligosaccharides by this enzyme. Acetolysis of the oligosaccharide with a molecular weight of 1720 released the radioactivity as mannobiose and mannotriose suggesting that these oligosaccharides contain di- and trisaccharide branches linked to the main chain in 1 leads to 6 linkages.


Thermodynamics of Ca2+ binding to troponin-C. Troponin-C (TnC) from rabbit skeletal muscle contains two high affinity Ca2+ binding sites (sites 1 and 2) that bind Mg2+ competitively (Ca2+-Mg2+ sites) and two Ca2+ binding sites (sites 3 and 4) of lower affinity that do not bind Mg2+ (Ca2+-specific sites). The free energy (deltaG0i), enthalpy (delta H0i) and entropy (deltaS0i) of binding Ca2+ to each of the four sites (i = 1 to 4) on TnC have been evaluated from microcalorimetry and equilibrium dialysis. The enthalpy of Ca2+ binding to each site was identical (-7.7 kcal mol-1); the entropy of Ca2+ binding to sites 1 and 2 was deltaS01,2 approximately equal to 14.7 e.u. whereas delta S03.4 approximately equal to 8.0 e.u. The positive entropy associated with Ca2+ binding to sites 1 and 2 is probably due to displacement of water produced by the alpha-helix formation, known to accompany the binding of Ca2+ to the Ca2+-Mg2+ sites. Thus, Ca2+ binding to the Ca2+-Mg2+ sites is driven by both enthalpy and entropy and the lower Ca2+ affinity for sites 3 and 4 is reflected in the lower entropy of Ca2+-binding. The entropy associated with Ca2+ binding to sites 3 and 4 suggests that some change in protein conformation is occurring upon binding of Ca2+ to these sites.


Congenital dislocation of the hip. The relationship of premanipulation traction and age to avascular necrosis of the femoral head. A retrospective study involving 276 patients with 319 congenitally dislocated hips was undertaken to assess whether premanipulation fraction prevents avascular necrosis. Roentgenographic criteria were utilized and only cases of frank dislocation unassociated with other disorders were included. It was found that in a child less than one year old, premanipulation traction of twenty-one days or more substantially reduces the incidence of avascular necrosis. For the child more than a year old, however, the prognosis is not as good, even if premanipulation traction is prescribed for twenty-one days or more. We found a 14 per cent incidence of avascular necrosis in children less than three months old and suggest that the cartilaginous femoral head prior to the appearance of the ossific nucleus is particularly vulnerable to vascular injury.


Wrist arthrodesis in paralyzed arms of children. A new technique for fusion of the wrist in children six years old and older is described. An iliac graft is sandwiched in a coronal split in the radius and carpal bones. Thirty-four children, most of them suffering residual paralysis from poliomyelitis, were treated with this method. It was successful in thirty-two patients, and these children had improved function of the extremity. They were followed to maturity; shortening of the forearm averaged 4.1 centimeters.


Early arthrodesis for a flail shoulder in young children. In seven children, five to nine years old, with a flail shoulder and a functional hand and elbow, arthrodesis of the shoulder in 80 to 90 degrees of abduction was performed by denuding the humeral head and glenoid of cartilage and fixing the humerus to the glenoid with Steinmann pins first inserted in the humerus in a proximal-distal direction and then driven in the reverse direction into the glenoid. Postoperatively each limb was immobilized in a spica cast, the pins were removed after a month, and the cast was removed after ten to twelve weeks when union was evident. All seven fusions united and function was much improved. When these children were followed to adult life there was only a small loss in humeral length and no change in position of the fused shoulder. Early arthrodesis is advocated in the treatment of a flail shoulder in children.


Instability of the subtalar joint. Diagnosis by stress tomography in three cases. Lateral instability in the subtalar joint was documented by a method of stress tomography in three patients. The subtalar inversion angle averaged 38 degrees +/- S.D. 6 degrees in asymptomatic feet and 57 degrees +/- S.D. 5 degrees in those with unstable subtalar joints . Review of the literature suggests that as many as 10 per cent of patients having the Watson-Jones procedure for abnormal talar tilt also have instability in the subtalar joint. It is felt that the subtalar joint should be evaluated in patients seen with the clinical symptom of lateral instability.


Ewing's sarcoma -- functional effects of radiation therapy. Long-term morbidity was assessed in fifty-five patients with Ewing's sarcoma who survived two years or longer. Primary site, radiation dosage, and age of the patient all had an important bearing on the end result. Guidelines for treatment of the primary tumor were developed. Patients with upper-extremity lesions were generally best treated by radiotherapy. Young children and some of the older patients with lesions in the lower extremity were generally best treated by primary amputation, depending on the site of the lesion.


Cervical orthoses. A study comparing their effectiveness in restricting cervical motion in normal subjects. The effectiveness of five cervical orthoses in restricting cervical motion was evaluated quantitatively in normal subjects using roentgenograms and overhead photographs made at the extremes of three planes of motion, while the effectiveness of the halo with a plastic body-vest was studied in seven patients with cervical fractures or local fusions. Flexion and extension were measured at each cervical intervertebral joint and combined cervical motion was measured for rotation and lateral bending. The best conventional braces restricted only 45 per cent of flexion-extension at the atlanto-axial joint; the halo restricted 75 per cent. The conventional braces were more effective in the middle and lower portions of the cervical spine. The results may prove to be useful guidelines for the selection of an appropriate orthosis to control motion in different planes and at different levels of the spine.


Ipsilateral hip and knee replacements as a single surgical procedure. A total of thirteen surgical procedures were performed on eight patients, all of whom had a history of rheumatoid arthritis. Five patients had bilateral hip and knee replacements and three patients had ipsilateral hip and knee replacements for a total of twenty-six joint replacements. Ambulatory status for all patients was significantly improved and all patients subjectively reported a great reduction in pain postoperatively. Complications included two transient peroneal-nerve palsies in the same patient from which she recovered spontaneously, and one questionable pulmonary embolus which resolved with heparin therapy. The relative advantages of this type of procedure and the indications for and against surgery are discussed.


A longitudinal study of the radiolucent line at the bone-cement interface following total joint-replacement procedures. After 100 consecutive total hip arthroplasties, a radiolucent line at the bone-cement interface as seen on serial roentgenograms was on the acetabular side in fifty-nine hips and on the femoral side, in three. Similarly, after seventy-five consecutive non-hinged knee-replacement arthroplasties, a radiolucent line was found at the bone-cement interface beneath the polyethylene component more frequently in knees in which the more constrained geometric implant was used (forty-two of fity-nine) than in knees in which the less constrained polycentric and modular prostheses were used (three of sixteen). The presence of a lucent line at the bone-cement interface in these joints did not signify looseness or failure of the arthroplasty.


Fractures of the scapula. An analysis of forty cases and a review of the literature. In forty patients treated at The University of Texas Medical Branch Hospitals for fifty-two fractures of forty-one scapulae from 1961 through 1973, most of the fractures were the result of vehicular accidents. The thirty-eight fractures involving only the body, neck, or spine of thirty scapulae (without involvement of the acromion, glenoid, or coracoid process) were successfully treated with a sling and early active motion. Three other fractures involving body were associated with three fractures involving the glenoid. The eleven fractures of the acromiom, glenoid, or coracoid process resulted in loss of motion in ten of the eleven shoulders. For fractures of this type we now recommend immobilization in 60 degrees of abduction, 25 degrees of flexion, and 25 degrees of external rotation, with early active abduction exercises. Open reduction is rarely indicated, only when there is disruption of the glenohumeral joint.


Flexible (silicone) implant arthroplasty of the metacarpophalangeal joint of the thumb. Forty-four thumbs in thirty-four patients with rheumatoid arthritis were treated with a flexible implant arthroplasty of the metacarpophalangeal joint. After follow-ups ranging from two to six and a half years the results were rated good to excellent in forty-two thumbs since they were not painful, had an increased arc of motion of the metacarpophalangeal joint in a more functional range, and were more effective in the activities of daily living. The results in this series indicated that careful reconstruction of the extensor apparatus is essential, as well as stabilization of the interphalangeal joint either by arthrodesis or by tenodesis if there is a hyperextension deformity. Of the six patients who had a fusion of the metacarpophalangeal joint in one thumb and an arthroplasty in the other, five preferred the arthroplasty because the joint was stable and also had a useful arc of motion, while one noted no difference between the two thumbs.


Fracture healing in rat femora as affected by functional weight-bearing. Femoral fractures were created in rats to determine whether there were differences in healing under conditions of immobilization and under conditions of immediate weight-bearing. Histological and roentgenographic differences were present by the second week after fracture and differences in mechanical properties were also present. These differences became progressively greater during the next three weeks. Functional weight-bearing was found to accelerate the rate of fracture healing and to improve significantly the strength of the healing bone.


The effect of fixation with intramedullary rods and plates on fracture-site blood flow and bone remodeling in dogs. Of fifty-seven adult dogs in which standard bilateral mid-ulnar fractures were produced, thirty-four had one side fixed with either a tight-fitting intramedullary Steinmann pin or a four-hole plate while the other side was not fixed, and twenty-three had a plate on one side and an intramedullary rod on the other. The blood flow at the fracture site, as determined by 85Sr clearance, was significantly less on the rod-fixed side than on the plate-fixed side at fourteen and ninety days, while the total ulnar blood flow was significantly higher on the rod-fixed than on the platefixed side at one and fourteen days but was the same on both sides thereafter. When the plate-fixed and rod-fixed fracture sites in the same dog were compared with respect to periosteal and endosteal bone formation as determined by tetracycline labeling anc microradiography, bone formation, like the blood flow, was greater in the callus of the ulnae fixed by plates. However, at ninety days, when healing had advanced to a point when the extent of union could be judged clinically, union had occurred in both ulnae of six of the seven dogs with both types of fixation studied at this time and was delayed in both ulnae of the remaining dog.


Roentgenographic search for avascular necrosis of the head of the femur in alcoholics and normal adults. Roentgenograms of 1,410 hips from 705 alcoholics were compared with those of 200 hips from 100 controls for changes consistent with non-traumatic (idiopathic) necrosis of the femoral head according to the findings diagnostic of necrosis of bone that were outlined by the Decompression Sickness Panel of the British Medical Research Council. Abnormalities were found in 45.3 per cent of the alcoholics and 49 per cent of the controls. None of the findings showed any statistical difference between the controls and the alcoholics. It was concluded that the roentgenographic findings used were not valid in detecting the early or subtle changes of necrosis of the femoral head and that the changes found in both groups are common and probably non-specific.


Tensile strength of wire-reinforced bone cement and twisted stainless-steel wire. To assess the tensile strength of wire-reinforced bone cement as used for posterior spine fusion, standardized bone-cement specimens reinforced with stainless steel and Vitallium wires 0.5 and one millimeter in diameter were tested in tension. The results showed that tensile strength of bone cement was increased significantly by reinforcing it with metal wires, the increase in strength being proportional to the number of wires used. Even after failure of the cement, the reinforcing wires still carried an appreciable amount of load, thus avoiding the catastrophic failure of the cement alone. Specimens reinforced with Vitallium wires in general failed at higher loads than specimens reinforced with similar stainless-steel wires. Pull-out studies showed that a hook or a loop at the embedded end of straight wires significantly increased the load necessary to pull the wires out. The tensile strength of twisted stainless-steel wire composed of two 0.5-millimeter strands increased with the number of turns up to about eight turns per inch (2.54 centimeters) and then decreased.


[By pass of the femoro-popliteal axis by autogenous saphenous graft. 100 by passes with a follow up between 1 and 5 years (author's transl)]. Venous by pass of the femoro-popliteal axis is very useful in certain special indications. A study of 100 by pass operations with a follow up between 1 and 5 years showed the indications and technic. A study of secondary worsening and comparison between three attitudes: abstinence, restoratives reoperations and preventive reoperations showed the superiority of routine detection of asymptomatic lesions and their preventive treatment. In cases of thrombosis of the bypass, a study of the general condition will permit one to choose between abstinence and a complex treatment including successively thrombolysis and then surgical removal of the cause of the thrombosis.


[Reflux into the pancreatic duct during peroperative biliary radiomanometry. Report of 200 cases (author's transl)]. The authors have observed 200 cases of reflux into the pancreatic duct out of 2,500 biliary operations with radiomanometry. Out of the 200 cases, 109 were functional, i.e. due to a common duct with or without an ampoule of Vater, or due to duodenal stockage whether there was a common duct or not. They had no therapeutic consequence. 38 were organic due to an obstruction (impacted gall stone, or odditis which favours reflux) and needed sphincterotomy especially when the pancreatic duct was dilated. In 53 cases, no organic or anatomical explanation was found. Reflux occurring at physiological pressures is more likely to be functional. The degree of reflux does not always depend on the increase in perfusion pressure. The time of the reflux depends partly on its nature. Early reflux is found mainly in cases of obstruction. Reflux secondary to passage into the duodenum is mainly found in functional reflux with stockage. Wirsung's duct is dilated in more than 75% of cases with obstruction and in only 3 cases out of 20 with functional reflux. Reflux into Santorini's duct is 3 times more common when there is papillary obstruction.


[Treatment of the post-thrombotic syndrome: modification of May's femoral by-pass (author's transl)]. During May's femoral by-pass, one should avoid by sapheno-popliteal anastomosis, obstruction of the femoral vein damaged by the post-thrombotic syndrome. After the early work of Kunlin, arterio-venous fistulas are often found in reconstructive venous surgery. By making a transverse end-to-side anastomosis between the main peripheral saphenous vein and the posterior tibial artery behind the internal malleolus, one may obtain a protective effect on the sapheno-popliteal anastomosis, increasing the blood flow towards the main saphenous vein across the perforating veins down to the deep leg veins. The protective effect of the arterio-venous fistula seems to be of interest during inactivity of the muscular pump of the calf (postoperative phase and during rest at night).


[Interest and limits of arteriography in one case of abdomino-pelvic trauma (author's transl)]. Hepatic and pelvic arteriography were carried out in one case of trauma of the liver and pelvis. Initial hepatic arteriography did not give more information than per-operative cholangiography. Secondary hepatic arteriography suggests the presence of lesions which, in fact, do not exist. Pelvic arteriography permitted us to localise on one obturator artery, the origin of a voluminous retroperitoneal hematoma. We were able to stop the hemorrhage by embolisation and this facilitated surgical removal of the hematoma.


[Rational use of antibiotics in digestive surgery]. The abusive and inappropriate use of antibiotics in digestive surgery has led to the selection of multi-resistant strains responsible for sometimes dramatic infective complications. The study of the normal digestive flora under pathological conditions, knowledge of the mechanisms of resistance, led us to propose the use of antibiotics with circumspection, depending on the germ isolated and the probable efficacy of the drugs. Prophylactic antibiotic therapy is only justified with regard to gram positive anaerobic bacteria or, in exceptional cases, where the risk seems high and may irremediably compromise the surgical procedure. Curative antibiotic therapy, after treatment of the infective focus, is often sufficient but depends on the bacteria: Whether anaerobic, staphylococci of Gram negative bacilli, with regard to which one should use narrow spectrum single antibiotics.


[Resistance and biological tolerance of six "inert" parietal protheses. Experimental and critical study (author's transl)]. Six different materials (3 mesh and 3 cloth) commonly used for the repair of abdominal wall hernias were evaluated in rats. 490 animals, distributed among four experimental groups, were implanted with one of the different prostheses through musculature and peritoneum. Serial macroscopic and bacteriological investigations were done. Using an original device, bursting strength of the wound was determined in all groups at intervals. Histological criteria were used to characterize the resistance of the wound and the tolerance of the host to foreign material. Statistical analysis of the results demonstrated that: 1) after the 15th post-operative day, the resistance of the wound was similar for each material tested; 2) during this early period mesh materials exhibited more resistance to bursting pressures than cloth materials; 3) the incorporation of mesh material was constant whereas encystment or extrusion was always observed after implantation of cloth material; 4) no infection occurred with mesh material but significant bacteria were found in 18 p. 100 of cloth material implantations; 5) the extent of the cellular reaction, the enumeration of giant, inflammatory cells and fibroblasts showed superiority of mesh material; 6) the ratio of fibroblasts/inflammatory cells reflected closely the mechanical resistance and tolerance of the foreign material.


Trace analysis of diethylstilbestrol [DES] in animal chow by parallel high-speed liquid chromatography, electron-capture gas chromatography, and radioassays. An analytical method is described for determining residues of the synthetic estrogen diethylstilbestrol (DES) in animal chow at levels as low as 1 ppb. A methanol extract of the chow is subjected to a 3-step cleanup procedure including a Sephadex LH-20 column, liquid-liquid partitioning at pH 14 and 10.2, and a silica gel column. Residues of DES in the cleaned-up extract are analyzed directly by high-speed liquid chromatography or derivatized to pentafluoropropionyl-DES and assayed by electron-capture gas chromatography. Tests with 14C-DES were used to develop and validate the procedure. Ancillary data concerning extraction efficiencies of various solvents, comparisons of various derivatizing reagents, rates of trans-cis isomerization in two solvents, P-values of DES in various solvent-systems and thin-layer chromatographic behavior i


Separation of aza-arenes by high-pressure liquid chromatography. Fast, efficient liquid chromatographic separations of aza-arenes were obtained using both, reversed-phase, and adsorbent packings. Aza-arenes with 2-5 rings are separated within 20 minutes. Sample recovery is quantitative and permits subsequent uv and fluorescence spectrophotometric identifications. The detection limit for most aza-arenes was 1 ng with a 254-nm uv detector. An application to an air pollution problem demonstrates the usefulness of this approach.


Selective osmoreceptor dysfunction in the syndrome of chronic hypernatremia. A patient with the syndrome of chronic hypernatremia (serum Na+: mean = 154, range 139-184 mEq/l, n = 30) and hypodipsia due to a hypothalamic injury was studied to evaluate osmolar and baroreceptor control of arginine vasopressin (AVP) secretion. Resting plasma AVP levels measured by radioimmunoassay were inappropriately low for the degree of plasma hyperosmolality: range = less than 0.5-2.1 pg/ml, n = 10, with corresponding levels of plasma osmolality (P osM) greater than 300 m osmol/kg, suggesting either direct damage to the AVP synthesis and storage area or impaired afferent osmoreceptor function. Direct pituitary damage seemed unlikely, since anterior pituitary function was normal by standard testing. The existence of adequate neurohypophyseal stores of AVP was demonstrated by baroreceptor stimulation with the hypotensive agent trimethaphan (Arfonad): plasma AVP rising to 50.0 pg/ml during transient hypotension (BP = 70/0). Osmoreceptor function was evaluated during acute water loading followed by hypertonic saline infusion. During hypertonic saline infusion plasma AVP levels correlated with P osM (R = .87, P less than .01, n = 8), suggesting some residual osmotic regulation of AVP release. The osmotic threshold for AVP release (the x-axis intercept of the plasma AVP-P osM regression line) was not higher than normal. However, the AVP levels throughout this study remained markedly subnormal for the degree of plasma hyperosmolality (maximum plasma AVP = 1.9 PG/ML when P os M = 327 M OSMOL/KG). Since a substantial amount of AVP was released with baroreceptor stimulation, the inadequate rise in plasma AVP level with hyperosmolality indicates that afferent input from the osmoreceptor/thirst area of the hypothalamus is selectively impaired in this patient. These findings directly demonstrate a dissociation of osmoreceptor function from the AVP secretory apparatus in man.


Plasma 1,25-dihydroxyvitamin D levels in patients receiving anticonvulsant drugs. Recent evidence has linked altered plasma vitamin D metabolite levels to the reported occurrence of hypocalcemia and other metabolic abnormalities in patients receiving anticonvulsant drugs. We have measured plasma levels of 25-hydroxyvitamin D (25-(OH)D) and 1,25-dihydroxyvitamin D (1,25-(OH)2D) in institutionalized patients on diphenylhydantoin (Dilantin) and/or phenobarbital therapy. Values were compared with those obtained in institutionalized patients receiving no drugs and with normal ambulatory subjects. Although plasma 25-(OH)D levels were lower in the patients on drugs, a deficiency of 1,25-(OH)2D, the tissue active metabolite of vitamin D, was not present. These results indicate that in patients taking anticonvulsant drugs, the serum calcium, phosphorus, alkaline phosphatase and parathyroid hormone (PTH) abnormalities are not caused by a defective formation of 1,25-(OH)2D.


Catecholamine excretion in "idiopathic" edema: decreased dopamine excretion, a pathogenic factor? In 16 women with idiopathic edema, urinary dopamine excretion was decreased when compared to control women (146 +/- 13 SE ng/ml/m2 vs. 212 +/- 32, P less than 0.05 in the supine position and 140 +/- 9 vs. 199 +/- 20, P less than 0.005 combined values of supine and recumbent positions) and was also lower when pooled values for urinary dopamine excretions both before and after furosemide were compared in idiopathic edema patients and in control subjects (270 +/- 30 ng/ml vs. 480 +/- 70, P less than 0.05). These patients have lower basal sodium excretions, decreased tubular rejection fractions of sodium in the upright position and lower urinary sodium excretions following furosemide administration. The urinary sodium and dopamine excretions before and following furosemide are positively correlated in control (P less than 0.05), idiopathic edema patients (P less than 0.02) and in both groups combined (P less than 0.005). Idiopathic edema patients have normal urinary noradrenaline and adrenaline excretions but, as previously observed, elevated: 1) plasma renin activity while either recumbent or upright, and 2) plasma aldosterone concentrations while upright. These results suggest that a decrease in urinary dopamine, a catecholamine recently recognized to have natriuretic action, possibly reflects a suppression of the renal dopaminergic system and may contribute to the excessive sodium retention in idiopathic edema either directly or indirectly through the renin-aldosterone system.


Studies in the biotransformation of cortisol to the cortoic acids in man. II. The central role of tetrahydrocortisol and tetrahydrocortisone as intermediates. The possible role of THF and THE as intermediates in the biotransformation of cortisol to the cortoic acids was studied by giving 3H-THF + 14C-cortisol tracers and 3H-THE + 14C cortisol tracers to two subjects each, measuring the 3H/14C isotope ratios of the urinary cortoic acid metabolites and relating these ratios to the dose ratio. Isotope ratios substantially higher than the dose ratio indicate that the tetrahydro compound is a better precursor than cortisol, and isotope ratios that are essentially identical to that of urinary THF or THE, respectively, indicate that the tetrahydro compound may be an obligatory intermediate in the cortisol leads to cortoic acids pathway. The isotope ratio data in these studies clearly establish that THF was a preferential precurosor of the 11 beta-hydroxy cortoic acids (cortolic and beta-cortolic) and THE was a preferential precursor of the 11-ketone cortoic acids (cortolonic and beta-cortolonic). Furthermore, the data strongly suggest that THF and THE may be obligatory intermediates in cortoic acid formation.


Inhibition of intestinal glucagon-like immunoreactivity (GLI) secretion by somatostatin in man. This work was undertaken to investigate the effect of somatostatin on intestinal glucagon-like immunoreactivity (GLI) secretion in man. In normal subjects GLI release is slightly stimulated by oral glucose while this sugar evokes a much greater GLI response in gastrectomized patients. Therefore, our study was performed in a group of such patients (N = 6). As expected, in the control experiments glucose ingestion elicited a clear-cut elevation of GLI plasma levels as measured with two antisera, 78J and R-8 (maximal peaks: 340% and 150% above basal values, respectively). Somatostatin infusion did not modify fasting GLI concentrations but completely abolished GLI response to glucose. Termination of the infusion was followed by a rebound of circulating GLI. The well-known suppressor effect of somatostatin on glucagon and insulin secretion was also detected. Finally, during somatostatin infusion the initial elevation of blood sugar after oral glucose, in the absence of insulin response, appeared considerably delayed. Our data demonstrate that somatostatin behaves as a potent inhibitory agent of GLI secretion in man. A retarding effect of somatostatin on glucose absorption is also compatible with our results.


Localization of estrogen synthetase in the chorionic villus fraction after homogenization of human term placenta. In previous investigations of human placental estrogen synthetase (aromatase), activity was found in the microsomal and mitochondrial fractions after homogenization and subcellular fractionation of placental tissue in isotonic sucrose. We report here that the majority of the placental aromatase sediments in a much heavier particulate fraction than the microsomal or mitochondrial fractions. After low speed centrifugation of the placental homogenate, only about 20% of the homogenate aromatase activity was measured in the supernatant, the precursor of the microsomal and mitochondrial fractions. Even after extensive homogenization in high speed shearing-type blenders and tight-fitting glass homogenizers the majority of the placental aromatase (more than 70%) consistently sedimented at very low centrifugal forces (100 x g ro 10 min) independent of whether the suspending medium contained up to 1.2M sucrose, or IM NaCl, or 50% glycerol. By selectively separating components of the low speed pellet via a sieving procedure or manual dissection of placental tissue, we found that the majority of the enzyme activity is associated with placental chorionic villus fragments. Several extensively washed preparations of villus fragments with aromatase specific activities similar to that found in placental microsomes were synthesis in the villus aromatase fraction was determined to be 1beta,2beta-H removal which is identical with that by placental microsomes and human ovaries. Evidence for the presence of cytochrome P-450 in the villus fraction was obtained by measuring the carbon monoxide-dithionite difference spectrum.


The metabolic response to moderate exercise in diabetic man receiving intravenous and subcutaneous insulin. The responses to moderate exercise of circulating energy fuels and endocrine pancreatic hormones were examined in insulin-dependent diabetics receiving insulin either sc or by continuous iv infusion. Eight subjects received one-third of their usual daily insulin doses sc in the thigh 1 h prior to exercise. Seven subjects exercised during infusion (iv) of insulin at 8-20 mU"min, started 12-14 h earlier. Exercise was on a bicycle ergometer for 45 min at 50% maximum oxygen consumption. The diabetics receiving sc insulin showed a sharp decline in glycemia from elevated resting levels (227 +/- 16 mg/dl), in contrast to the control subjects whose glycemia did not change. The control subjects insulin (IRI) fell, and glucagon (IRG) remained unchanged. In the sc-insulin diabetics, exercise induced a further rise in IRG from elevated levels (296 +/- 76 pg/ml). Resting lactate, pyruvate and alanine were normal and increased as in controls. Though FFA, glycerol and ketone body levels were normal at rest, FFA failed to rise with exercise as in the controls and glycerol and ketone body increments were smaller. RQ increased and remained elevated in contrast to the later fall in controls during exercise. These results are consistent with selective insulin deficiency at rest, and increased insulin effect during exercise. This resulted in greater carbohydrate utilization during exercise, but without the normal shift back toward utilization of fat-derived fuels with continuation of exercise. Diabetics receiving insulin by infusion showed no glycemic change with exercise. Exercise caused greater increases in lactate and pyruvate levels (4-fold), although alanine levels increased only during recovery. The significantly elevated resting FFA levels showed a rise which was sustained at higher than control values during recovery; glycerol and ketone body increments also tended to be greater than in controls. Intravenous insulin sustained euglycemia in exercise, obviating the fall in glycemia with sc insulin. The responses of other metabolite levels were abnormal, and consistent with a subtle degree of underinsulinization.


Hormonal evaluation of the intrauterine progesterone contraceptive system. Nine women were studied for one menstrual cycle prior to the insertion of an intrauterine progesterone contraceptive system (IPCS) delivering 65 microng progesterone/day into the uterus and again at 1 month after its insertion. Eight of these women were again studied between 6-8 months after the insertion of the IPCS. Luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol-17beta, progesterone, prolactin and relaxin were measured in each plasma sample. The data from each study were combined according to the day of the LH peak. Ovulation occurred in all the cycles studied in spite of an elevation in plasma estradiol-17beta and a depression of prolactin and relaxin immunoactivities at the 6-8  month follow up. Menstruations noted at the 6-8 month of use occurred while levels of estradiol-17beta and progesterone were elevated.


Extrathyroidal conversion of thyroxine to 3,3',5'-triiodothyronine (reverse-T3) and to 3,5,3'-triiodothyronine (T3) in humans. In order to estimate the relative magnitude of the two alternative pathways of monodeiodination of thyroxine (T4) in adult humans, the metabolic clearance rates (MCR) and production rates (PR) of 3,3',5'-triiodothyronine (reverse-T3,rT3) and of 3,5,3'-triiodothyronine (T3) were determined in six euthyroid control subjects (C) and in five hypothyroid patients (H) receiving L-T4 as replacement therapy (0.15-0.3 mg/day). MCR was computed by a non-compartmental method of analysis from the plasma disappearance of 125I rT3 and 131I T3 during 72 h following simultaneous injection of tracers. PR was calculated from MCR and the serum concentration of rT3 and T3, respectively, determined by radioimmunoassay. In the H subjects, rT3 MCR averaged 97.1 +/- 12.8 (SD) 1/day and rT3 PR, 34.3 +/- 12.8 microng/day; T3 MCR was 28.7 +/- 6.1 1/day and T3 PR, 20.3 +/- 6.6 microng/day (all corrected to 70 kg body weight). These results were not significantly different from those in the control group; rT3 MCR 104 +/- 24 1/day, rT3 PR 33.0 +/- 9.2 microng/day; T3 MCR 24.0 +/- 5.9, T3 PR 24.2 +/- 4.1. The proportionof total triiodothyronine (rT3 averaged 62% in H patients and was similar (57%) in the C group. The results obtained in the H subjects indicate that the production of rT3 is a major route of T4 metabolism, equal to or exceeding that of T3. From the close agreement between the mean values for rT3 PR in the C and H groups it is concluded that most, if not all of the rT3 produced in normal humans is derived by extrathyroidal conversion from T4.


The effects of temperature and plasma cortisol on distribution of aldosterone between plasma and red blood cells: influence on metabolic clearance rate and on hepatic and renal extraction of aldosterone. Aldosterone enters red blood cells (RBC) to a greater extent at 37 C than at lower temperatures. The ratio of 3H-aldosterone concentration in RBC to that in plasma increases from 0.2 at 4 C to 0.7 at 37 C when cortisol concentration is low. Increasing plasma cortisol increases the RBC/plasma aldosterone ratio. When plasma transcortin (CBG) is saturated with cortisol, the RBC/plasma ratio of 3H-aldosterone approaches 0.93, the ratio observed in RBC incubated in 4% albumin solution. The effects of plasma cortisol and temperature on the RBC/plasma ratio reflect an affinity of aldosterone for plasma CBG greater than the affinity for plasma albumin or RBC. Hepatic extraction averages 92% of plasma and RBC aldosterone. Neither hepatic extraction nor renal extraction (less than or equal to 20%) is significantly altered by changing plasma cortisol concentration. Whole blood MCR of aldosterone is unaffected by redistribution of aldosterone from plasma to RBC when plasma cortisol increases, but both plasma cortisol and the temperature at which blood is separated affect the RBC/plasma ratio of 3H-aldosterone and thus change the calculated plasma MCR. The RBC transport of aldosterone, and its dependence on temperature and plasma cortisol, must be taken into account in the evaluation of plasma aldosterone concentration.


Action of metergoline in suppressing prolactin release induced by mechanical breast emptying. Ten women were studied during the first post-partum week. Mechanical breast emptying with a breast pump for 30 min induced a marked increase in plasma prolactin (PRL) reaching a maximum 30 min after the beginning of stimulation. In all patients oral administration of the antiserotonin agent metergoline, 8 mg daily for two days, completely blocked any PRL secretion induced by breast stimulation. These results confirm the inhibitory effect of metergoline on PRL release.


Effect of methysergide and pimozide on apomorphine-induced growth hormone secretion in men. The dopamine receptor agonist, apomorphine HCl (0.75 mg sc) increased serum growth hormone (GH) concentrations in each of 6 subjects. This increase was not inhibited by pretreatment with the serotonin antagonist, methysergide, but was completely abolished in the 4 individuals pretreated with pimozide, a selective dopamine receptor blocking agent. These results provide further evidence that apomorphine-induced GH secretion is mediated by a dopaminergic mechanism and is independent of a serotonergic mechanism.


Inadequate corpus luteal function in women with benign breast diseases. The corpus luteum function of 109 patients with benign breast disease was appreciated by way of plasma progesterone and estradiol determinations during the luteal phase. These patients had ovulatory cycles according to a biphasic basal body temperature curve; blood samples for plasma progesterone and estradiol estimation were collected between the first and the last day of the thermal plateau following the thermal nadir. Results obtained were compared to those observed in 50 normal ovulatory women. In the patients' group, the mean daily levels for progesterone ranged from 3.5 +/- 0.4 (SE) ng/ml to 8.1 +/- 3.8 (SE) ng/ml according to day of blood collection. These values are significantly different from the corresponding daily values observed in normal women. No significant difference was observed concerning plasma estradiol between patients and normal women. These findings indicate that women with benign breast disease have an inadequate corpus luteum function which may be secondary to an ovulation disorder. Pathophysiological implications resulting from this observation are discussed.


Transient fall and subsequent return of high aldosterone secretion by adrenal adenoma during continued dexamethasone administration. Plasma aldosterone concentration was consistently decreased by 50% or more in 6 patients with aldosterone-producing adenoma on the first day of dexamethasone administration, only to rise subsequently with continued use of dexamethasone while plasma cortisol concentration remained suppressed. The secondary rise in plasma aldosterone was not related to measured changes in known stimuli of aldosterone secretion. It is probable that the observations result from intrinsic alteration of aldosterone synthesis in the adenoma during prolonged ACTH suppression.


Overnight gonadotropin excretion in normal females. FSH and LH in acetone precipitates of timed overnight urine collections were measured by radioimmunoassay. FSH and LH urinary excretion extrapolated to 24 h was determined in 140 normal girls 3-16 years. Timed overnight urines were collected throughout a menstrual cycle from 3 normal adult women. FSH and LH progressively increased with age from 2.0 +/- 0.5 (FSH) and 2.0 +/- 0.4 (LH) IU/24 h for the 3--4 yr age group to 8.1 +/- 1.8 (FSH) and 11.5 +/- 3.8 (LH) IU/24 h for the 15--16 yr age group. Mean FSH and LH excretion for succeeding age groups (3--4, 5--6, 7--8, 9--10, 11--12, 13--14 and 15--16 years) increased significantly for the 5--6, 9--10 and 11--12 year age groups. Mean LH but not FSH excretion increased significantly for the 13--14 year age group. FSH and LH excretion increased progressively with stage of sexual development. In the 3 girls with established menstrual cycles. FSH (43.6 +/- 5.0 IU/24 h) and LH (113.7 +/- 7.8 IU/24 h) peaks occurred on the same day. Luteal phase excretion of FSH and LH was significantly lower than follicular phase excretion. Timed overnight urine collection allows integration of several hours of FSH and LH excretion.


Irradiation of the head. Immediate effect on growth hormone secretion in children. Plasma growth hormone (GH) was determined in samples obtained hourly from 1000 h to 0700 h before, and after a 3 week course of "prophylactic" cranial irradiation, in ten leukemic children who had no clinical or laboratory evidence of central nervous system involvement. The mean per hour value of GH prior to irradiation (4.1 +/- 2.4 ng/ml) was similar to that of  7 endocrinologically normal children (4.09 +/- 2.4 ng/ml), a finding strongly suggesting normal pituitary function prior to irradiation. Following irradiation GH levels at each hour were, in general, lower than before. The difference is statistically significant at 0100 h. The peak GH responses were lower following than preceeding irradiation (p less than 0.02). The findings suggest an immediate suppressive effect of irradiadiation on the apparently normal hypothalamic pituitary axis in children.


Ultrastructural localization of pancreatic polypeptide in human pancreas. Human pancreatic polypeptide (HPP) was localized in surgically obtained human pancreas by immunocytochemical electron microscopy. HPP was localized to the secretory granules of cells distinct from the alpha, beta, or delta cells of endocrine islets. The PP cells in 10 patients surveyed were most frequently located in the periphery of the islets. Immunolocalization distinguishes a fifth endocrine cell type in human islets which distinct ultrastructurally by its smaller granule diameter and lack of PP immunoreactivity.


An abnormality in steroid reductive metabolism in a hypertensive syndrome. Studies in a juvenile hypertensive syndrome associated with suppressed plasma renin activity and hypokalemic alkalosis failed to reveal overproduction of aldosterone or any other known steroid. There was however an abnormal increase in the fraction of unconjugated urinary steroids. Analysis of this fraction following the administration of labeled cortisol revealed that it was largely composed of dihydro metabolites reduced either at 4,5 or at C-20 and that the 4,5-dihydro fraction contained an abnormal increase in 5alpha- relative to 5beta-metabolites. There was, however, no absolute defect in the complete reduction of ring A to form tetrahydro derivatives. These findings, together with observations by Marver and Edelman that 5alpha-dihydrocortisol may be an effective mineralcorticoid, suggest the possibility of an etiologic relationship between the metabolic abnormality and the patient's hypertensive disorder.


Normal insulin binding to cultured fibroblasts from patients with lipoatrophic diabetes. Confluent fibroblasts were assayed for insulin binding to determine whether there was an inherent abnormality in receptor function to explain the insulin resistance of lipoatrophic diabetes (LD). Cells from three patients and four controls were compared for confluent density, receptor saturation, specific and non-specific binding and the concentration of unlabelled hormone producing 50% competition for binding with labelled ligand. Cells from patients with LD did not differ significantly in any of these characteristics from the controls. These findings indicate that there is not a basic defect in insulin receptors of LD patients. A secondary disruption of binding, e.g. by a circulating factor, remains possible.


Pharmacokinetics of alcohol following single low doses to fasted and nonfasted subjects. The absorption and elimination characteristics of alcohol have been studied in healthy fasted and nonfasted human volunteers using low single doses. In non-fasted subjects, carbohydrate reduced overall alcohol bioavailability by about 96 percent, compared to 90 per cent for fat and 75 per cent for protein. Inhibition of absorption in nonfasted subjects appeared to be due to less alcohol being available for absorption rather than a reduced absorption rate. Serum alcohol levels in fasted subjects were interpreted in terms of both first-order and zero-order absorption followed by first-order elimination. Of the two proposed models, that utilizing zero-order absorption provided a marginally better fit to observed data.


Antihypertensive and renin angiotensin effects of metolazone with and without propranolol. Renin angiotensin system parameters and blood pressure (B.P.) were followed monthly in patients with essential hypertension on metolazone, 5 mg daily for three months and with added propranolol, 40 to 160 mg, for the subsequent three months. On metolazone alone at three months, sitting B.P. declined from 166/108 +/- 14/11 mm Hg to 145/98 +/- 14/9 mm Hg (P less than 0.005). Plasma renin activity (PRA) increased from 3.9 +/- ng/ml/hr to 10.4 +/- 8.6 ng/ml/hr (P less than 0.005); plasma angiotensinogen did not change. Venous blood angiotensin I and II levels (pg/ml) rose initially but returned toward control values. A significant decline in plasma renin substrate reactivity (PRSr) in index occurred. Propranolol addition caused further lowering of only systolic B.P. and predominantly in the standing position, more marked at one month (40 mg) than at three months (160 mg). No significant further changes were observed in any of the measured parameters of renin angiotensin system, except for a rise in PRSr index concomitant with B.P. elevation at three months. Metolazone-induced changes in B.P. showed significant correlations at three months with changes in PRSr index. It is concluded that during chronic metolazone administration, the overall activity of the renin angiotensin system was diminished or unchanged. Propranolol did not inhibit metolazone stimulated PRA but did cause further decline in B.P. in the first two months, unrelated to renin angiotensin system.


Pharmacokinetics of cefazolin in patients with renal failure; special reference to hemodialysis. The pharmacokinetic constants of cefazolin were determined comparatively in ten normal subjects, 12 patients with renal failure, and ten patients on repeated hemodialysis. Significant correlations could be established between the creatinine clearance values and the overall elimination rate constants (Ke), serum half-lives (T 1/2), plasmatic areas (Ftot), urinary recoveries (uo-6 hr), and renal clearances of the antibiotic (Cr). The pharmacokinetic changes produced by extrarenal removal were also studied. On the basis of the obtained results, a dosage schedule adjusted to renal status was proposed.


Bioavailability of a sustained-release dyphylline formulation. The bioavailability of dyphylline from sustained-release tablets, studied in seven normal subjects given a single dose of the drug, was 67.8 per cent of that from conventional tablets. In three subjects given 40 mg/kg dyphylline as sustained-release tablets every 8 hours for four days, serum dyphylline concentrations were maintained above 7.88 microng/ml once steady state was achieved. No adverse effects were noted, though in one subject peak serum dyphylline concentrations reached 30 microng/ml after multiple doses.


Long-term diuretic therapy with metolazone of renal failure and the nephrotic syndrome. The effects of the new diuretic metolazone were studied in ten patients with chronic renal insufficiency and ten with nephrotic syndrome. Patients were maintained on metolazone for up to 44 months. Beneficial effects of treatment included loss of edema and improved control of blood pressure. The natriuretic effect of metolazone facilitated the use of sodium bicarbonate to treat acidosis in several patients. Concurrent administration of metolazone and furosemide produced a dramatic diuresis in one patient resistant to either diuretic alone. Adverse effects of metolazone therapy were those characteristic of other effective diuretics, Including serum electrolyte losses and hyperuricemia. Initial treatment produced small increases in serum creatinine among patients with renal insufficiency, suggesting that GFR was decreased secondary to diuresis-induced volume depletion. The study demonstrates that metolazone is both safe and effective over long periods of time.


Anti-keyhole limpet hemocyanin antibody in normal unsensitized individuals. Antibody to keyhole limpet hemocyanin (KLH) has been detected in normal unimmunized subjects by means of a sensitive micropassive hemagglutination technique. Prior sensitization was not detected by either skin testing or lymphocyte transformation. After immunization with KLH there was no correlation between the level of this antibody and subsequently acquired skin test reactivity and lymphocyte transformation.


Ultrastructural identification of mast cells obtained from human bronchial lumens. We have investigated the morphology of mast cells in human bronchial lavage fluid. Under light microscopy, these cells were seen to contain discrete metachromatically positive granules that were indistinguishable from those present in mast cells and basophils from other sources. With electron microscopy, the bronchial mast cells were shown to be mononucleated and to contain granules which primarily consisted of particulate materials. Complex structures such as scrolls and crystalloids were also found in the granules. In order to delineate the relationship between bronchial mast cells and mast cells from other anatomic sites, we studied the ultrastructure of tissue mast cells from bronchial, lung parenchymal, and bone marrow biopsy specimens, and that of basophils from peripheral blood. There were similarities as well as differences in these cells from different sources--in their size, shape, lobes of nuclei, nuclear chromatin distribution, and structure of special, presumably histamine-containing, cytoplasmic granules. Based on the electron microscopic data, we believe the intralumenal cells previously described in human beings can be classified as mast cells. The presence of viable, immunoreactive mast cells in the lumen of bronchi may have a significance in relation to respiratory allergic reactions, especially the immediate-type respiratory response.


Quantitative requirements for C3 to induce Forssman systemic shock and cutaneous hemorrhagic vasculitis in guinea pigs. The requirements of complement (C) to induce systemic and cutaneous Forssman reactions were studied in inbred DHC-BA and Hartley strain guinea pigs. After intravenous injection of Forssman antibody, fatal systemic shock was associated with a marked drop in CH50, C4, and C3 and a lesser decrease in C5 hemolytic activity. Platelet counts and leukocyte counts dropped as well. With the use of the purified low molecular weight factor from cobra venom (CVF) to deplete C3, guinea pigs with less than 1% intravascular C3 were protected from lethal shock. Approximately 1% to 3% C3 activity is required for Forssman cutaneous vasculitis. These results confirm earlier studies that classical complement pathway activation occurs in Forssman shock and demonstrate the exquisite biologic efficiency of C3 in provoking the shock syndrome.


Melittin: an allergen of honeybee venom. The presence of serum IgE antibodies to melittin was tested by the radioallergosorbent test (RAST). Melittin, the principal protein of honeybee venom, was isolated by gel filtration on Sephadex G-75 and covalently bound to cyanogen bromide-activated microcrystalline cellulose. The melittin preparation was homogenous by immunoelectrophoresis with the use of rabbit antiserum to whole honeybee venom and by polyacrylamide electrophoresis in gels containing 1% sodium dodecyl sulfate. Elevated serum IgE antibodies to melittin (three times greater than binding by normal sera) were found in 7 of 24 honeybee venom-sensitive persons and in 5 of 20 nonsensitive beekeepers. In one venom-sensitive patient a particularly high titer of IgE antibody was found. The reaction between solid-phase melittin and IgE antibody could be inhibited by fluid-phase melittin but not by phospholipase A (PLA). Similarly, the reaction of IgE antibody with solid-phase PLA was inhibited by PLA but not by melittin. In passive transfer skin tests with the sensitive patient's serum, positive wheal-and-flare reactions were obtained in 3 nonallergic recipients following melittin challenge; appropriate controls were negative. These results indicate that melittin is an allergen in some honeybee venom-sensitive patients and in an occasional patient melittin may be a major allergen.


A simple method for keeping allergenic extracts at a temperature near 4 degrees C during in-office use for skin testing and immunotherapy. A method is needed for keeping ragweed pollen aqueous allergenic extracts at a temperature near 4 degrees C while they are in use for skin testing and immunotherapy in order to prevent deterioration in potency from warming. A polystyrene foam icebox is described which can be cooled with sacks of coolant which have been reduced to a temperature of - 17 degrees C in the freezing compartment of a standard refrigerator. Both icebox and coolant can be readily obtained from standard commercial sources and are inexpensive. When sufficient adequately cooled coolant is properly employed, this icebox will maintain the temperatures of allergenic extracts in the desirable range of 2 degrees to 4 degrees C while they are in use for skin testing and immunotherapy.


Function and histology of testes from aged coturnix maintained on different photoperiods. The prolonged effects of short, medium, and long photoperiods on longevity, cloacal gland size and function, and the testicular morphology of the aged coturnix were studied. In all testes, the effects of aging were characterized histologically by decreased tubular sperm concentration, increased abnormal sperm and cellular debris, as well as dislodgement of maturing sperm from Sertoli cells. However, testicular abnormality in the long photoperiod quail was more pronounced than in the short photoperiod quail whose testes were judged to be morpholoigcally in better condition, thus indicating an aging-photoperiod relationship. Aging had little or no effect on the cloacal gland function, although its size similar to the testicular histology, was affected by different photoperiods. Cloacal gland size and function can indicate sexual potency, but not necessarily the reproductive capabilities of aged coturnix. Also, an increased incidence of cataract formation was recorded in aged coturnix, unrelated to photoperiods.


Physiological characteristics of a champion runner: age 77. A world champion distance runner, age 77, was evaluated in various anthropometric, pulmonary, hematological, and cardiorespiratory measures. The subject held 14 recognized world records in track and distance running for competitors aged 74, 75, and 76 years and had 11 records pending for the 77-year-age category. He was the 1975 National AAU Master's class IV-b champion in the 400, 800, 1500, 5000 and 15,000 meter races. The results of the physiological measures taken exceeded those of a more sedentary population of comparable age, were similar to reported values for other Master athletes, and approached the standards noted for much younger runners. The largest age-related decrements were observed in measures of maximal heart rate, maximal pulmonary ventilation, and maximal oxygen consumption. The general reduction in aerobic power was attributed to a combination of pulmonary, circulatory, and metabolic factors in addition to training consideration.


Patterns of menopause: a study of certain medical and physiological variables among Caucasian and Japanese women living in Hawaii. Comparisons were made between menopausal women and nonmenopausal controls among Caucasians and Japanese living in Honolulu, to investigate the extent of physical changes and clinical symptoms associated with menopause. The analysis was conducted using the multiphasic screening records of 170 menopausal cases and 162 nonmenopausal controls in Caucasians, and of 159 menopausal women and 187 nonmenopausal controls in Japanese. Discriminant function analysis was employed with relevant anthropometric, medical, and physiological variables. After adjusting for the linear and non-linear effects of age, only surgery and medication were retained as significant discriminant variables. Discriminant functions for Caucasian and Japanese groups were not found to be significantly heterogeneous. With regard to the discrimination of the menopausal and nonmenopausal groups, the data suggested that, while no clinical conditions other than those attributable to the effects of aging were significantly associated with the menopausal state, medication and surgical procedures for female disorders were significantly related to menopause.


Lithium treatment in the elderly. I. Clinical usage. Clinical use of lithium salts in manic states in the elderly is reviewed. Special problems with toxicity are discussed. A dose titration method that has been safely used for 10 years is presented and illustrated with case material. Lower blood levels than usually advised are recommended.


Sociological and social-psychological correlates of anomia among a random sample of aged. In this investigation the distribution of responses on an anomia index (Srole's anomia scale) was examined for a national probability sample of the elderly. Step-wise multiple regression procedures were used to determine the relative importance of the independent variables in explaining the variation of anomia. Of the zero-order correlations, the "sociological" variables, e.g., education, work status, occupational prestige, and yearly income, were the strongest correlates of anomia. The social psychological variables, viz., various attitudinal and perceptual phenomena, also exhibited reasonably strong correlations. Those variables labeled "activity/organizational," e.g., various indices of social engagement, revealed the lowest correlations. In step-wise regression analysis six statistically significant explanatory variables were, in descending order: (1) a judgement of one's relative financial standing, (2) city size, (3) race, (4) work status, (5) education, and (6) occupational prestige.


Change in life satisfaction: a longitudinal study of persons aged 46-70. Longitudinal analyses were performed on changes in life satisfaction among a sample of 378 community residents, aged 46-70. There were no significant changes in the mean life satisfaction scores for any age-sex cohort and no significant differences between the sexes, although there were some individual changes. Life satisfaction at the end of a 4-year-period was significantly related to initial levels of self-rated health, social actiivty, and sexual enjoyment. Attempts to predict residual change in life satisfaction by initial levels or by changes in these variables were largely unsuccessful.


Relationship of siblings to the elderly person's feelings and concerns. This study investigated the feelings and concerns of 64 elderly persons, as revealed by a projective instrument, in relation to number of sex of siblings. Partilal correlations, with effects of age, education, and occupation removed, were computed separately for each sex. The number and proportion of female siblings were found to have a greater influence than male siblings on the elderly's feelings and concerns. For men, sisters appeared to be emotionally supportive, while for women sisters appeared to be more challenging, as indicated by greater social concerns.


Relocation of the aged: a review and theoretical analysis. The literature on the relocation of the aged is examined and the findings are presented within a framework of three types of moves--institution to institution, home to institution, and home to home--with each type having a voluntary and involuntary component. A theoretical model is proposed to explain the often contradictory results reported in this literature. In brief, it is argued that an individuals' response to relocation is largely determined by (a) the perceived predictability and controllability of the events surrounding a move and (b) differences in controllability between pre- and post-relocation environments. The validity of this model is tested by examining existing research and suggestions for future research are made.


Interpersonal skill training with institutionalized elderly patients. A behavioral program to train elderly nursing home patients in interpersonal skills was systematically developed and evaluated. Data obtained from intensive interviews with patients, staff, and professionals were used to construct a training program and self-report and behavioral measures for assessing the program's effectiveness. Twenty-five patients were randomly assigned to the training program or to one of two control condotions. On a behavioral measure of responses to situations that had been included in training, the Skill Training program was superior to the control conditions. However, learning failed to generalize to novel situations. Directions for future skill training research are discussed.


Foster placement for the older psychiatric patient. The effect of age on foster care outcome was examined for 572 male psychiatric patients referred for foster care from five VA hospitals. Subjects were assigned randomly to preparation (experimentals) or continued inpatient care (controls.) Research staff collected  data on social functioning, mood, activity, and adjustment. Only 79 (14%) were 60 years or older. Compared with younger patients, those 60 and over were more likely to be diagnosed as having chronic brain disease and less likely to be diagnosed as have schizophrenia. About 70% of the younger and older experimentals were placed into foster care. Alcoholics and persons with more hospitalizations were significantly less likely to be placed. Patients in foster care changed significantly compared with hospitalized controls by having less social dysfunction and better adjustment. Older subjects improved as a result of being seen in the hospital prior to placement but were not different from younger subjects in how they responded to the foster home. The study suggests that foster care is an appropriate and underutilized resource for elderly psychiatric patients.


Recall of specific suppression: co-dominance of suppression after primary or secondary antigen stimulation. Mouse spleen cells primed in vivo with beta-galactosidase (GZ), and cultured in vitro with trinitrophenyl beta-galactosidase (TNP-GZ), make a very poor anti-TNP response compared to cultures of normal spleen cells. The lack of response is caused by active suppression as indicated by cell mixture experiments. At 2-months following priming, while the helper effect predominates, suppression can be recalled by rechallenge of the mice with GZ. Both primary suppression (after a single injection) and secondary suppression (after two injections) reduce the response of normal or carrier-primed helper cells at equivalent cell ratios in mixture experiments. Both kinds of suppression are carrier specific, slectively suppress high avidity antibody production, and require cell division to suppress the response of normal cells. The only difference between primary and secondary suppression is that primary suppression is obliterated in cultures which are not challenged with antigen until 24 hr after cultures are established. Secondary suppression is still observed when cultures receive TNP-GZ at 24 hr. Suppression appears to be an early component of both primary and secondary responses. The regulatory function of such suppression may be the delay of antibody secretion.


Membrane transport by murine lymphocytes. II. The appearance of thymidine transport in cells from concanavalin A-stimulated mice. We have investigated membrane transport of thymidine and adenosine by bulk murine nonadherent spleen cells from animals treated with the lectin concanavalin A. Separation of cells from radioactive medium was achieved by means of the oil microfuge technique which permits incubation periods as short as 4 sec. We were able to distinguish between the membrane-linked function of transport and uptake which includes subsequent accumulation and metabolism of the transport solute. Previously we reported that cells derived from untreated mice could not be shown to translocate thymidine in a carrier-mediated fashion. In contrast, cells from concanavalin A-treated mice showed two membrane transport systems for thymidine, a high affinity system (Km = 160 micronM) and one with low affinity (Km = 4 mM). The transport of thymidine conformed to the usual criteria for membrane carrier-linked function: increased uptake with time, substrate saturability and chemical specificity. In contrast to thymidine uptake, the transport of adenosine by cells from lectin-treated mice was similar to that shown by cells from untreated animals. Thus a specific membrane-linked activity is altered differentially in the course of mitogen-effected lymphocyte stimulation. We have found that determination of DNA synthesis by the standard method gave values which were as much as 700% too high, since the counts obtained by direct precipitation with TCA of cells incubated with radio-labeled thymidine exceed the cell-associated radiolabel obtained by the rapid sampling technique. With lectin-stimulated cells, the discrepancy observed was up to three times greater. Hence the validity of the standard assay for blastogenesis must be viewed with caution.


The biologic activity of affinity cross-linked oligomers of rabbit immunogloblin G as tested by antigody-dependent cell-mediated cytolysis (ADCC). The abilities of normal spleen cells and macrophage-like and lymphocyte-like tumor cells to serve as effectors in antibody-dependent cell-mediated cytolytic (ADCC) reactions were examined. It was observed that those cells which contained Fc receptors on their surfaces were effective mediators of ADCC whereas those not bearing Fc receptors were inactive. Oligomers of IgG were able to inhibit the ADCC reaction and, with the macrophage line, inhibition and binding closely paralleled each other. With the lymphocyte lines, a large degree of inhibition of ADCC occurred when only small amounts of oligomers were bound; with normal spleen cells inhibition occurred in the same range of oligomer concentrations required to inhibit the tumor-mediated ADCC reactions. The data suggest that the interaction between antibody-coated erythrocytes and effector cells is unexpectedly weak.


The role of an activatable esterase in immune-dependent phagocytosis by human neutrophils. Diisopropylphosphofluridate, cyclohexyl alkylphosphonofluoridates, and cyclohexyl phenylalkylphosphonofluoridates, which are potent, irreversible inactivators of serine esterases, inhibit the phagocytosis of opsonized sheep erythrocytes by human neutrophils. Two types of inhibition were observed: a) 'cell-dependent' inhibition, which is determined by measuring the ingestion of neutrophils pre treated with the esterase inhibitors and washed; and b) 'phagocytosis-dependent' inhibition, which is due to the presence of the inhibitors during phagocytosis. With the cyclohexyl alkylphosphonofluoridates, phagocytosis-dependent inhibition was always greater than cell-dependent inhibition. Cell-dependent inhibition was irreversible and dependent on the duration of the incubation of neutrophils with inhibitor. Both types of inhibition were dependent on the concentration of the inhibitor. Poorly or non-phosphorylating analogues of the cyclohexyl alkylphosphonofluoridates of DFP were not inhibitory; nor did fluoride, the hydrolysis product of these inhibitors, inhibit ingestion under either condition. In addition, neither method of treating the neutrophils resulted in a decrease in neutrophil viability. Furthermore, pretreating the EAC1423 with the inhibitors did not decrease ingestion. We conclude that cell-dependent inhibition is due to the inactivation of an esterase required for phagocytosis which is in or on the neutrophil in an active form, and thus is susceptible to inhibition by the esterase inactivators before contact of the neutrophil with the phagocytic stimulus. Phagocytosis-dependent inhibition is interpreted as being due to inactivation of an esterase required for phagocytosis which is normally in an inactive precursor proesterase form that is activated by the interaction of the neutrophil with the phagocytic stimulus. The distinctly different inhibition profiles of the active and activatable esterases indicate that they are two different activities.


Hypogammaglobulinemia in experimental myeloma: the role of suppressor factors from mononuclear phagocytes. We have investigated the pathogenesis of the polyclonal hypogammaglobulinemia associated with BALB/c plasmacytomas TEPC-183 and SPQC-11 to gain insight into the hypogammaglobulinemia observed in human myeloma. With pokeweed mitogen-driven IgM biosynthesis by mouse splenocytes as the indicator system for suppression, we found that a protein extract of asscites cells obtained from these tumor-bearing animals could suppress immunoglobulin production, whereas like extracts from a non-suppressing plasmacytoma, modified RPC-5, caused no suppression in vitro. Extracts of tumor ascites depleted of mononuclear phagocytes by iron carbonyl treatment showed little suppressor activity. The active extract was not cytotoxic and contained no mycoplasma or common murine viruses. Furthermore, the active suppressor factor appears to be a low m.w. protein that is not affected by treatment with ribonuclease. These results and others are consistent with the idea that the hypogammaglobulinemia of myeloma is due to the formation of immunoregulatory macrophage-like cells which synthesize a suppressor substance.


Isolation and enumeration of peripheral blood monocytes. Monocytes were isolated by counterflow contrifugation from human peripheral mononuclear cells obtained from whole blood by the Ficoll-Hypaque technique. Examination of volume spectra of the monocytes showed a bimodal distribution with peaks at 330 and 370 micron3. The upper peak contained 61.4 +/- 3.3% of the total monocyte population, whereas the remainder were in the lower peak which overlaps the lymphocyte spectrum. The purity of the monocyte preparations from six normal adult donors, identified by peroxisomes observed by electron microscopy, was 90.4 +/- 1.6%.


The reactions of antibodies to paragloboside (lacto-N-neotetraosyl ceramide) with human erythrocytes and lymphocytes. Paragloboside (lacto-N-neotetraosyl ceramide) is a biosynthetic precursor of the ABH and P1 blood group glycosphingolipids and of one class of gangliosides. This study concerns the specificity of rabbit anti-paragloboside antibodies, andtheir reactions with human erythrocytes and lymphocytes. Lacto-N-neotetraose was approximately 10 times as effective as lacto-N-tetraose in inhibiting complement fixation by purified IgG antibodies to paragloboside. The purified antibodies cross-reacted weakly with lactosyl ceramide, asialo GM1, and lacto-N-neotetraosyl polylysine. The purified IgG and IgM antibodies exhibited weak hemagglutinating activity which was enhanced by treatment of erythrocytes with papain or neuraminidase. In contrast to a macroglobulin described by Tsai et al. (J. Immunol. 117:717, 1976), which also binds paragloboside and lacto-N-neotetraose, anti-paragloboside antibodies reacted equally well with normal adult, O1, and umbilical cord erythrocytes. The IgG antibodies to paragloboside reacted with about 60% of B lymphocytes of peripheral blood and 5 to 10% of T cells. The relationship between antibodies to paragloboside and other lectins and antibodies directed against beta-galactosyl groups, including anti-T (Thomsen-Friedenreich) antibodies, is discussed.


Use of bovine conglutinin for the assay of immune complexes. We have used purified bovine conglutinin to develop a solid state radioimmune assay for immune complexes. Employing aggregated human IgG incubated with fresh normal human serum (complement) as a model for immune complexes, we have shown that our conglutinin assay is sensitive and highly specific for immune complexes that have fixed complement. The assay preferentially detects large complexes, is minimally influenced by monomeric IgG, and can be inhibited by high ionic strength, calcium chelation, and acetamido sugars. In addition, we have surveyed several hundred clinical sera from patients with various immunopathologic disorders and find evidence of immune complexes in some.


Effect of cytochalasin B on human lymphocyte responses to mitogens: time and concentration dependence. Human peripheral blood mononuclear cells were exposed to various concentrations of concanavalin A (Con A) and cytochalasin B (CB). Low concentrations of CB (0.5 to 2.0 microng/ml) augmented, whereas high concentrations of CB (greater than 2.0 microng/ml) inhibited the mitogenic response to Con A when Con A and CB were added at the initiation of culture. Augmented responses could also be found by adding normally inhibitory high concentrations of CB after 72 to 96 hr incubation with Con A. Preincubation of the cells with CB 5 min to 4 hr before adding Con A also resulted in potentiation of the mitogenic response. CB did not increase spontaneous transformation and worked as well in various solvents (dimethyl sulfoxide, acetone, and ethanol). The effect was totally reversible by removing CB before adding Con A. CB had no effect on 3H-Con A binding to cells. The potentiating effect of CB was still evident after removal of CB, Con A, or both from cultrues that had been simultaneously exposed to Con A and CB for at least 2 days. CB augmented the response of human lymphocytes to Con A-adsorbed, x-rayed autologous lymphocytes. In the presence of optimal concentrations of CB, x-ray-damaged cells respond better to Con A; the response to suboptimal and supraoptimal concentrations of Con A is better, and the response with delayed addition of alpha-methyl-D-mannoside is better. CB also potentiates the response of human lymphocytes to pokeweed mitogen and phytohemagglutinin. Direct observation of cells in culture demonstrated the development of huge clusters of cells during mitogenic responses to Con A in the absence of CB. Circumstances which inhibited the mitogenic response to Con A totally prevented the development of cellular aggregates (prolonged incubation with high concentrations of CB). Each of the conditions which augmented the mitogenic response to Con A was accompanied by the formation of multiple small, relatively uniform cellular aggregates. These observations suggest that CB affects cell-cell interaction possibly by affecting distribution of Con A on the cell surface, by affecting cell sruface net negative charge, or by increasing the response to chemotactic stimuli. CB may be acting in some manner to enable certain nonresponsive cells to respond to a mitogenic stimulus, although an increase in the response of individual cells cannot be ruled out.


Interferon production in lymphocyte cultures after rubella infection in humans. When stimulated with rubella virus irradiated with ultraviolet light, cultures of lymphocytes from individuals immune to rubella responded with production of 12-14 times more interferon than did cultures of lymphocytes from individuals susceptible to rubella. This property may be retained throughout life, since it was seen in individuals who had had rubella in their recent and remote pasts. Peak levels of interferon were detected six days after stimulation of lymphocytes with rubella virus antigen. In contrast, phytohemagglutinin induced production of interferon equally well in cultures of lymphocytes from persons immune and persons susceptible to rubella, with levels peaking three days after stimulation. Since there was a positive correlation among titers of hemagglutination-inhibiting antibody to rubella virus, incorporation of [14C]thymidine, and production of interferon after stimulation of lymphocyte cultures by rubella virus, interferon synthesis may be considered to be another measurable parameter of cell-mediated immunity in rubella.


Influenza B-associated Reye's syndrome: incidence in Michigan and potential for prevention. Prospective surveillance for Reye's syndrome in Michigan revealed the occurrence of 46 cases between December 15, 1973 and June 1, 1974. In an attempt to determine the incidence of influenza B-associated Reye's syndrome, a randomized point-prevalence survey of 1,041 schoolchildren was done in a county in Michigan where there had been simultaneous outbreaks of influenza B and Reye's syndrome. Of the children tested, 20% had titers of hemagglutination-inhibiting antibody to influenza virus B/Hong Kong/5/72 of greater than or equal 1:20. Based upon this countywide survey, the incidence of Reye's syndrome following influenza B was estimated as between 30.8 and 57.8 cases of Reye's syndrome per 100,000 cases of influenza B. A detailed epidemiologic investigation of the patients who developed Reye's syndrome indicated that the syndrome occurred four times more frequently in children living in rural areas than in children in the urban areas of the state. These studies indicate that, in addition to antecedent viral infections such as influenza B, factor(s) that are most likely extrinsic or environmental also play a role in the pathogenesis of Reye's syndrome.


Toxoplasmosis and the host-parasite relationship in murine schistosomiasis mansoni. Infection with Toxoplasma gondii has been shown to suppress markedly the in vitro splenic lymphocyte response to nonspecific mitogens and the in vivo antibody response to sheep red blood cells. The effect of toxoplasmosis on an in vivo cell-mediated response, granuloma formation around Schistosoma mansoni eggs, was examined in the present study. The granulomas were markedly suppressed from two to 20 weeks after infection with T. gondii. In subsequent experiments the effect of toxoplasmosis-induced immunosuppression on the development of hepatosplenic schistosomiasis was evaluated. Mice with the combined infections had markedly smaller hepatic granulomas and lower mean portal pressures than those infected with S. mansoni alone. Although the prevalence of esophageal varices in the mice with schistosomiasis alone was 60%, there was no visible collateral circulation in the animals with both infections.


Quantitation of amphotericin B with use of high-pressure liquid chromatography. A chemical method for determination of concentrations of amphotericin B in serum and cerebrospinal fluid (CSF) is described. After extraction with methanol, the antibiotic was separated by reverse-phase, high-pressure liquid chromatography and quantitated by absorption at 405 nm. The lower limit of detection of this assay was 0.02 microng/ml. Relative standard deviations of less than 3.6% were noted for multiple determinations of sera containing 0.20 and 1.00 microng of amphotericin B/ml. No interfering peaks were found in extracts of serum or CSF from normal humans or in extracts of serum from patients treated with other drugs and antimicrobial agents, including 5-fluorocytosine. Comparison of the method with microbiological assays showed correlation coefficients of 0.90 and 0.83 for serum and CSF determinations, respectively. This chemical assay is very rapid (less than 30 min), sensitive, accurate, and specific and appears to be suitable for routine clinical use.


Dose-response relationship after immunization of volunteers with a new, surface-antigen-adsorbed influenza virus vaccine. Volunteers (15 per group) were given inoculations of various doses (5-400 international units [IU]) of surface-antigen-adsorbed influenza virus A/Port Chalmers/73 vaccine; this vaccine was prepared from purified virus hemagglutinin and neuraminidase antigens and adsorbed to alhydrogel. The titers of hemagglutination-inhibiting (HAI) and neuraminidase-inhibiting antibodies in serum after immunization showed a clear dose-response relationship. Thus, for volunteers given 400, 100, 25, or 5.0 IU of vaccine, the titers of HAI antibody in serum increased 174-, 32-, 12-, and eightfold, respectively. A similar dose response was observed for production of local neutralizing antibody. Immunization with A/Port Chalmers/73 virus vaccine also induced serum HAI antibody to influenza viruses A/Scotland/74, A/England/72, and A/Hong Kong/68; the increase in titers of antibody to these viruses corresponded directly to the degree of cross-reactivity between the virus tested and the vaccine virus. Four weeks after immunization, all volunteers were challenged with attenuated WRL-105 influenza virus. Evidence of viral infection was found in one, two, and two volunteers in each group of 15 subjects previously immunized with 400, 100, and 25 IU of vaccine, respectively; in six of the 15 volunteers given 5.0 IU of vaccine; and in 10 of 15 control subjects. The results suggest that equivalent immunity was induced in volunteers given greater than or equal to 25 IU of vaccine.


Diarrhea associated with clindamycin and ampicillin therapy: preliminary results of a cooperative study. Patients treated with clindamycin were monitored for development of diarrhea and colitis in a cooperative study at three hospitals in Edmonton, Toronto, and Winnipeg, Canada. Ampicillin-treated patients were matched with clindamycin-treated patients. Diarrhea occurred in 62 (18%) of 343 clindamycin-treated patients compared with 16 (5%) of 315 ampicillin-treated patients (P less than 0.001). Among the three hospitals, the rate of diarrhea associated with clindamycin therapy ranged from 10% to 23% compared with 0 to 10% for ampicillin therapy. The only identified risk factor in the development of diarrhea was increased age; clindamycin-associated diarrhea occurred in 18 (46%) of 39 patients greater than or equal to 60 years old. No correlation was observed between the risk of diarrhea and the duration of treatment, total dosage of drug, route of administration, or type of underlying disease. Pseudomembranous colitis was diagnosed in seven (2%) of 343 patients treated with clindamycin and in one (0.3%) of 315 patients treated with ampicillin.


Gastrointestinal side effects of clindamycin and ampicillin therapy. Hospitalized patients who received clindamycin or ampicillin were evaluated for gastrointestinal side effects for a period of up to six weeks after therapy was discontinued. Of 104 patients receiving clindamycin therapy, 31 (29.8%) developed diarrhea, and two (1.9%) developed pseudomembranous colitis (PMC). Of 138 patients receiving ampicillin, 24 (17.3%) developed diarrhea, and one (0.7%) developed PMC. Diarrhea persisting for three days or more was noted in 13 (12.5%) of the patients receiving clindamycin and in seven (5.1%) of those receiving ampicillin. The tendency to develop diarrhea was positively correlated with serious illness, abdominal or pelvic sepsis, and total dosage of clindamycin. Examination of stools from a patient with PMC that was associated with clindamycin therapy showed a decrease in the number of anaerobic bacteria from the numbers found in stool cultures of normal controls. Those patients who did not develop diarrhea also had fewer anaerobic bacteria and coliform organisms. Lymphocytes from the patient with PMC were hyporeactive to phytohemagglutinin and hyperreactive to clindamycin.


Incidence of diarrhea and colitis associated with clindamycin therapy. A retrospective and prospective study was made of side effects in 200 patients who received clindamycin over a 15-month period. The incidence of gastrointestinal symptoms in this group of patients was 21.5%; of diarrhea, 13.5%; and of colitis, 2.5%. Factors associated with an increased incidence of diarrhea were older age and parenteral route of administration. Simultaneous use of other antibiotics, particularly aminoglycosides, and increased total dosage of clindamycin did not increase the incidence of diarrhea. The gastrointestinal side effects were usually self-limited if the drug was promptly discontinued and fluid support was provided.


Experimental infections with anaerobic bacteria in mice. Experimental anaerobic infections in mice are reviewed with a description of a model of pure Bacteroides fragilis infection. In this experimental situation, B. fragilis produces large subcutaneous abscesses in the groins of mice that can be diagnosed without autopsy. This infection was treated effectively with clindamycin in doses that produced levels of drug in blood similar to those attainable in humans.


Comparison of clindamycin and chloramphenicol in treatment of serious infections of the female genital tract. A study was performed of 102 obstetric-gynecologic patients who were thought to have sepsis or a pelvic abscess. Fifty-three of these women received chloramphenicol and 49 received clindamycin. In addition, all patients received penicillin or a similar antibiotic and an aminoglycoside. Similar clinical results were observed with the two treatment regimens. In eight of the 49 patients who received clindamycin and in three of 52 patients who received chloramphenicol, use of the drug was discontinued because of side effects. These combinations of antibiotics did not eliminate the necessity for major operative drainage, which was required in 40 patients. Resistant organisms were recovered from only two patients. Although sepsis and shock were most frequently associated with gram-negative aerobic bacteremia, they occurred in two patients in whom only anaerobes were recovered from blood cultures. Because the clinical results with the two regimens were equivalent, a decision to use either clindamycin or chloramphenicol should be based on the individual physician's assessment of the toxicity of these agents.


Techniques for anaerobic susceptibility testing. Minimal inhibitory concentrations (MICs) of antimicrobial agents for anaerobic bacteria can be determined by agar dilution and broth dilution (including microdilution) techniques. If MICs are not determined routinely, the disk broth or category methods are recommended for routine use. The Bauer-Kirby disk diffusion method and its interpretative standards should not be used for anaerobes.


Clindamycin and Carbenicillin in treatment of patients with intraabdominal and female genital tract infections. Clindamycin alone and with an aminoglycoside or carbenicillin alone and with an aminoglycoside were used in therapy for 173 patients with intraabdominal or genital tract infections. Excellent or good results were obtained in 115 of 131 patients treated with clindamycin and 33 of 42 patients treated with carbenicillin. Few serious side effects were observed in patients who received either drug. The data indicate that clindamycin is an effective drug in these conditions and suggest that carbenicillin may be equally effective.


In vitro activity and clinical efficacy of clindamycin in the treatment of infections due to anaerobic bacteria. Clindamycin, rosamicin, josamycin, and metronidazole had similar inhibitory activity against 29 clinical isolates of Bacteroides fragilis, i.e., 100% of strains were inhibited by 0.8 microng of metronidazole or josamycin/ml and 100% by 1.6 microng of clindamycin or rosamicin/ml. Metronidazole was bactericidal against 97% of the isolates, and clindamycin or rosamicin (in concentrations of 1.6 microng/ml) was bactericidal against 80%. Erythromycin and josamicin were the least bactericidal agents in vitro. Thirty-two patients with pleuropulmonary and intraabdominal or pelvic infections caused by anaerobic bacteria were treated with clindamycin. Cure was achieved in 27 patients. In another group of 37 patients treated with parenteral clindamycin, diarrhea developed in 30% and was significantly more common in those patients with abdominal or pelvic infection. Only one patient developed pseudomembranous colitis. These observations suggest that clindamycin is an excellent and relatively safe antibiotic for treatment of infections caused by anaerobes when combined with surgery or with other antibiotics selected for activity against aerobic gram-negative bacilli.