Regulation of phospholipid biosynthesis in isolated rat hepatocytes. Effect of different substrates. The effects of choline, ethanolamine and its N-methyl analogs, different fatty acids, and L-methionine on phospholipid biosynthesis via the CDP-ester pathways and the methylation pathway were studied in rat hepatocytes. Phosphatidylethanolamine synthesis was stimulated severalfold by 0.02 to 0.1 mM ethanolamine, especially in the presence of long chain unsaturated fatty acids. At higher concentrations of ethanolamine, phosphorylethanolamine accumulated but the level of CDP-ethanolamine and the rate of phosphatidylethanolamine synthesis did not increase further. The rate of phosphatidylcholine synthesis via the CDP-ester pathway responded in a way analogous to that of phosphatidylethanolamine synthesis upon the addition of choline and fatty acid, except that a 10- to 20-fold higher concentration of choline was required for maximal stimulation, probably due to the rapid oxidation of choline to betaine. Phospholipids containing N-monomethyl- or N,N-dimethylethanolamine were efficiently formed from the corresponding free bases in the absence of ethanolamine and choline. Ethanolamine, but not other bases, inhibited completely phospholipid formation from N-monomethylethanolamine, probably as a result of competition at the level of CDP-ester formation. The data indicate that the cytidylytransferase reactions are rate-limiting steps in the synthesis of phosphatidylethanolamine and probably also phosphatidylcholine. In addition, the availability of diacylglycerol and its fatty acid composition may significantly affect the rate of phospholipid synthesis. The rate of phosphatidylcholine formation via phospholipid N-methylation approximately doubled when L-methionine was added at concentrations similar to that in rat plasma. Under these conditions the rate of phosphatidylcholine synthesis via this pathway was 20 to 40 percent of that via diacylglycerols and CDP-choline. The methylation of phosphatidylethanolamine to phosphatidylcholine remained essentially constant when the rate of phosphatidylethanolamine synthesis was varied 8-fold, but was significantly reduced when the formation of N-monomethyl- or N,N-dimethylphospholipid was stimulated by addition of the corresponding base. These phospholipids not only replaced phosphatidylethanolamine as the substrate for methylation but also increased the rate of phosphatidylcholine formation via this pathway. A method for the determination of nanomole amounts of different ethanolamine compounds is described.


Chromatin changes during the cell cycle of HeHa cells. HeLa S3 cells were synchronized by selective mitotic detachment and chromatin was isolated from synchronized populations at various times during the cell cycle. The isolated chromatins were studied by circular dichroism and for their ability to bind the intercalating dye, ethidium bromide. Chromatin from cells in the middle of the S phase showed an increase in maximum ellipticity in the 250- to 300-nm region of circular dichroism spectra and an increased ability to bind ethidium bromide, when compared to chromatin isolated from mitotic cells. Chromatin from G1 cells had values intermediate between mitotic and mid-S chromatins.


The structure of rat proalbumin.

The structure of rat proalbumin. The structure of rat proalbumin, a liver precursor to rat serum albumin, has been determined to consist of the hexapeptide Arg-Gly-Val-Phe-Arg-Arg attached to the NH2 terminus of the polypeptide chain of rat serum albumin. Edman degradation of a proalbumin preparation for 14 rounds gave the major sequence Arg-Gly-Val-Phe-Arg-Arg-Glu-Ala-His-Lys-Ser-Glu-Ile-Ala. A comparison of cyanogen bromide fragments suggests that these two proteins differ only in this respect. On treatment with cyanogen bromide, these proteins gave three classes of peptides with molecular weights of 30,000, 10,000, and smaller than or equal to 5,000. A combination of gel filtration, electrofocusing, and ion exchange established that these peptides were indistinguishable, with exception of those of 10,000 molecular weight. By amino acid and sequence analyses this fraction from rat serum albumin was found to be the NH2-terminal fragment. Radiochemical amino acid and sequence analyses show that the NH2-terminal hexapeptide is the major fragment released from proalbumin by limited tryptic hydrolysis. The protein that remains cannot be distinguished from rat serum albumin.


Biosynthesis of dermatan sulfate. I. Formation of L-iduronic acid residues. L-[14C]Iduronic acid-containing sulfated galactosaminoglycans were formed by incubation of a fibroblast particulate fraction with UDP-D[14C]glucuronic acid, UDP-N-acetylgalactosamine, and sulfate donor (3'-phosphoadenylylsulfate). The formation of L-iduronic acid was strongly promoted by concomitant sulfation of the polymer. In the absence of sulfate donor 5 to 10% of the [14C]uronic acid residues were L-iduronic acid. However, when 3'-phosphoadenylylsulfate was included in the incubation mixture the amount of L-iduronic acid in the product increased 3 to 5-fold. Furthermore, approximately the same quantity of L-[14C]iduronic acid was recovered from the product formed in a pulse-chase experiment where incorporation of 14C-isotope preceded sulfation. It was therefore concluded that C-5 inversion of D-glucuronic acid to L-iduronic acid occurred on the polymer level as shown previously for the biosynthesis of heparin (Hook, M., Lindahl, U., Backstrom, G., Malmstrom, A., AND Fransson, L-A., J. Biol. Chem. (1974) 249, 3908). This conclusion was supported by the finding that no L[14C]iduronic acid could be detected in the UDP-hexuronic acid pool during this experiment. Nonsulfated and sulfated [14C]galactosaminoglycan products were degraded separately with chondroitinase-AC. The non-sulfated products afforded primarily disaccharide and a small amount of tetrasaccharide, while the sulfated products yielded, in addition, a considerable amount of larger oligosaccharides. Tetrasaccharides from nonsulfated products contained L-iduronic acid indicating that C-5 inversion at solitary sites can occur in the absence of sulfation of adjacent hexosamine moieties. The larger oligosaccharides obtained after chondroitinase-AC digestion of sulfated products yielded L-iduronic acid upon acid hydrolysis and were susceptible to chondroitinase-ABC digestion. The split products were almost exclusively 4-sulfated disaccharides. These results demonstrate that formation of blocks of L-iduronic acid-containing repeat periods is associated with 4-sulfation of adjacent hexosamine moieties.


Regulation and substrate specificity of a steroid sulfate-specific hydroxylase system in female rat liver microsomes. The sulfate-specific hydroxylase system in liver microsomes from rats has been investigated with respect to its substrate specificity. Eighteen different C18, C19, C21, and C27 steroid sulfates and the coresponding free steroids have been incubated with microsomal preparations from male and female rats. The sulfate-specific system was only present in preparations from female rats and primarily catalyzed hydroxylation in position 15beta but also in position 7beta. In contrast to this, male liver microsomes were more efficient than female liver microsomes in hydroxylating free steroids; these were hydroxylated in positions 2alpha,2beta,6alpha,6beta,7alpha,7beta,16alpha, and 18. The sulfate-specific hydroxylase system in female liver microsomes was found to have rigid requirements c concerning the structure of ring D in the substrate molecule; only 17beta-sulfates (C18 and C19 steroids) and 21-sulfates (C21 steroids) were hydroxylated. Less rigid criteria, however, exist concerning the structure of ring A. The following K-m values were determined for microsomal 15beta-hydroxylation: 5alpha-androstane-3alpha,17beta-diol disulfate, 17.2 muM; 5beta-androstane-3alpha,17beta-diol disulfate, 16muM;5alpha-androstane-3alpha,17beta-diol 17-sulfate, 26 muM; and estradiol 17-sulfate, 181 muM. Some of the regulatory mechanism controlling the activity of the sex-specific 15beta-hydroxylase system also have been studied and compared to the mechanism controlling the activities of the less specific 2alpha-, 7alpha-, and 18-hydroxylase systems active on 5alpha-[4-14C]androstane-3alpha,17beta-diol. Biliary drainage did not affect the 15beta-hydroxylase activity, whereas the 2alpha- and 7alpha-hydroxylase activities decreased..


Dihydrotestosterone formation in cultured human fibroblasts. Comparison of cells from normal subjects and patients with familial incomplete male pseudohermaphroditism, Type 2. The conversion of [1,2-3H]testosterone to [3H]dihydrotestosterone has been assessed in fibroblast monolayers grown from skin biopsies of foreskin, scrotum, and various nongential skins from 31 control men who varied in age from newborn to 25 years and three 46,XY subjects with hereditary male pseudohermaphroditism. Under the standardized conditions utilized in this study, the rate of dihydrotestosterone formation was greater in fibroblasts grown from genital skin (foreskin and scrotum) passages exhibit the same differentiation in dihydrotestosterone formation as the skin from which the fibroblasts were grown. Furthermore, 5alpha-reductase, the enzyme that converts testosterone to dihydrotestosterone, exhibits apparent similar substrate specificity in control foreskin fibroblasts and in the foreskin itself. Fibroblasts grown from the foreskin of two patients with familial incomplete male pseudohermaphroditism, type 2, an autosomal recessive disorder of phenotypic sexual differentiation, showed a marked deficiency in the capacity to form dihydrotestosterone. In contrast, fibroblasts grown from the scrotum of one 46,XY male with familial incomplete male pseudohermaphroditism, type 1, an apparent X-linked disorder of phenotypic sexual differentiation, formed dihydrotestosterone at a normal rate.


Characterization of the phosphoenolpyruvate inhibition of mitochondrial protein synthesis. The interaction of phosphoenolypyruvate with isolated rat liver mitochondria has been further investigated. The uptake of P-enolpyruvate is accompanied by the stoichiometric release of intramitochondrial adenine nucleotides resulting in the inhibition of mitochondrial protein synthesis. Addition of specific inhibitors of either the adenine nucleotide carrier or the tricarboylic acid carrier blocks the P-enolpyruvate-stimulated loss of adenine nucleotides and thereby prevents the resultant inhibition of mitochondrial protein synthesis. These data suggest that there is a specific interaction between the mitochondrial adenine nucleotide translocase and the tricarboxylic acid carrier resulting in the control of intramitochondrial adenine nucleotide levels of phosphoenolpyruvate.


Phosphate transport in rat liver mitochondria. Membrane components labeled by N-ethylmaleimide during inhibition of transport. N-ethylmaleimide (NEM) inhibits the transport of phosphate in mitochondria but is without effect on permeation of other metabolities. In spite of its specificity for inhibition of phosphate transport, NEM reacts in an unspecific manner with inner membrane proteins in general. Treatment of mitochondria with [3H]NEM just sufficient to produce inhibition of phosphate transport results in labeling of at least 10 polypeptide components of the inner membrane. A marked increase in the specificity of reaction of NEM for components of the phosphate transport system is attained by first protecting the transport system with p-mercuribenzoate (p-MB) and then by irreversibly blocking reactive sulfhydryl groups unassociated with transport by the addition of unlabeled NEM. Subsequent addition of dithiothreitol removes p-MB and restores 65 to 75 percent of the original phosphate transport activity. Reinhibition of transport with [3H]NEM results in both a 6-fold decrease in the amount of [3H]NEM bound by purified inner membrane vesicles and a substantial reduction in the number of labeled polypeptide components. Five distinct labeled species are detected by this method, one of which is a 32,000 molecular weight protein containing 40 percent of the bound radioactivity, or approximately 160 pmol/mg of inner membrane protein. Correlation of binding of [3H]NEM by inner membrane proteins with inhibition of phosphate transport suggests that the maximum concentration of the NEM-sensitive component of the phosphate transport system is 60 pmol/mg of mitochondrial protein. This value, when combined with V-max of NEM-sensitive transport of 205 nmol times min-1 times mg-1 at O degrees (Coty, W. A., and Pedersen, P. L. (1974) J. Biol. Chem. 249, 2593) yields an approximate minimum turnover for this process of 3500 min-1 at 0 degrees. This turnover number is at least 20-fold greater than similarly calculated values for adenine nucleotide transport and succinate oxidation in rat liver mitochondria at this temperature. Taken together these results suggest that the NEM-sensitive phosphate transport system in rat liver mitochondria has an unusually high catalytic activity compared to other mitochondrial processes, and that at least one of the five NEM-binding proteins is likely to be an essential component of this transport system.


Purified liver microsomal cytochrome P-450. Separation and characterization of multiple forms. During the purification of rabbit liver microsomal cytochrome P-450 (P-450LM), evidence was obtained for the occurrence of at least four distinct forms. These were distinguished by polyacrylamide gel electrophoresis after treatment with sodium dodecyl sulfate in the presence or absence of mercaptoethanol and were shown to have characteristic spectra as the reduced carbon monoxide complexes. They are designated by their relative electrophoretic mobilities. P-450LM2, which was purified to apparent homogeneity, is induced by phenobarbital and has a subunit molecular weight of 50,000. P-450LM4, which was also extensively purified, is induced by beta-naphthoflavone and has a molecular weight of 54,000. P-450LM1,7, which is induced neither by phenobarbital nor beta-naphthoflavone, is a mixtureMIXTURE OF ABOUT EQUAL AMOUNTS OF TWO FORMS WITH MOLECULAR WEIGHTS OF 47,000 AND 60,000 RESPECTIVELY. Some preparations were obtained containing primarily P-450LM1 or P-450LM7. Benzphetamine, ethylmorphine, and p-nitroanisole are hydroxylated preferentially by P-450LM2, and benzpyrene by P-450LM1,7. Biphenyl is hydroxylated in both positions 2 and 4 by all of the preparations, but the latter position is strongly favored by the action of P-450LM2. Testosterone is hydroxylated primarily in position 16alpha by P-450LM2 and in position 6beta by P-450LM1,7. Although the occurrence of additional forms of the cytochrome with highly similar electrophoretic behavior is not ruled out, it appears that the presence of these forms differing in subunit molecular weight may account for the variety of catalytic activities attributed to this pigment of liver microsomes.


Benign osteoblastoma: range of manifestations. A study of twenty-five new cases of osteoblastoma and a review of 172 previously reported cases demonstrated the wide spectrum of manifestations of the lesion. The ratio of males to females was 2:1, with 80 per cent of the patients being under thirty years of age. The duration of complaints prior to diagnosis averaged seventeen months. The majority of the lesions were found in the spine or major long bones of the lower extremity. When complete excision of the tumor was done a cure always was the result, but incomplete curettage also affected a cure in some cases. Three cases presented in detail emphasize the wide variation in biological behavior which this lesion may exhibit. Histological characteristics of aneurysmal bone cyst may frequently appear in the tumor tissue. Radiotherapy does not alter the course of the disease and appears to be contraindicated.


Ewing's sarcoma. A clinicopathological and statistical analysis of patients surviving five years or longer. Of 229 patients with primary Ewing's sarcoma of bone, thirty-seven survived five years or longer. When the patients who survived five years or longer were compared with the 192 who did not, two factors associated with longer survival were identified in this retrospective study: location of the primary tumor within an extremity and inclusion of surgery as part of the initial treatment. The probability of five-year survival for all 229 patients with primary Ewing's sarcoma was 16 per cent. For those seen since 1950, this probability was 18 per cent, and for patients without metastasis, it was 22 per cent.


Performance analysis of an ex vivo geometric total knee prosthesis. Studies were made of a well functioning geometric total knee prosthesis, removed at autopsy from a sixty-one-year-old man, which had been used for eight months. The components were firmly embedded in bone with no evidence of loosening. There was significant deformation of the polyethylene tibial component, but no significant wear on the metallic component. Thin layers of fibrous tissue were found in some areas of the bone-cement interfaces.


Restoration of strong opposition after median-nerve or brachial plexus paralysis. In hands in which the superficial flexors of the fingers and thenar muscles of opposition are denervated, usually as the result of high median-nerve or brachial plexus injury, thumb opposition can be restored by transfer of the superficial flexor of the ring finger to the thumb through a dynamic pulley made from the distal segment of flexor carpi ulnaris which is attached to the proximal segment of flexor carpi ulnaris to the transferred paralyzed superficial ring-finger flexor tendon. Of sixteen patients treated by this method, twelve had good or satisfactory results; one had a poor result; and three were lost to follow-up.


Gangrene of the newborn. A case report.

Gangrene of the newborn. A case report. Gangrene of the newborn is an uncommon condition usually resulting from decreased perfusion of a part, usually an extremity. There are a variety of situations which can result in this condition. Knowledge of the exact sequence of events which lead to the insult and the conditions surrounding it is helpful in determining the etiology and as a consequence the treatment of the lesion. In the patient described, direct pressure from the maternal pelvis was probably the contributing factor which led to venous occlusion of the extremity. Arterial thrombosis, emboli, trauma, congenital heart disease, sepsis, dehydration, coagulopathies, and venipuncture are other possible causes which should be considered. The treatment is in general supportive, allowing the ischemic area to demarcate and slough. Range-of-motion exercises and splinting to avoid contracture are helpful in the rehabilitative phase.


Fatigue fracture: the basic lesion is inthmic spondylolisthesis. The defect in the pars interarticularis in spondylolysis and spondylolisthesis is most often the result of repeated trauma, stress, and factors other than acute fracture. These fatigue fractures develop early in life, may have a strong hereditary basis, and most often represent incidental roentgenographic findings. Attention should be given to the youngster or adolescent with low-back pain and paraspinal muscle spasm. If these patients are followed closely, the incidence of pars interarticularis defect is higher than appreciated. The lesion in some of these individuals may progress to significant vertebral slipping. If the developing defect is recognized early, treatment can be quite satisfactory.


The treatment of trochanteric hip fractures using a compression screw. One hundred and four consecutive intertrochanteric hip fractures treated with a compression screw were reviewed. Despite death and lack of follow-up, the late results in sixty-two cases of fracture were analyzed. Three non-unions and one malunion gave the technique a failure rate of only 6.4 per cent. Twenty-two patients with both stable and unstable fractures were allowed to bear weight as early as tolerated, in an average of fourteen days. Early weight-bearing did not appear to compromise the end result.


The deep posterior compartmental syndrome of the leg. Fourteen cases of deep posterior compartmental syndrome of the leg complicating lower-extremity trauma were analyzed. Caused by increased pressure within the deep posterior compartment, the syndrome was characterized by pain, plantar hypesthesia, weakness of toe flexion, pain on passive toe extension, and tenseness of the fascia between the tibia and the triceps surae in the distal medial part of the leg. Decompression of the compartment within twelve hours of the onset of the syndrome prevented permanent sequelae.


Knee joint infections with Staphylococcus aureus and Micrococcus species. In rabbit knees 600 Staphylococcus aureus organisms produced progressive infection in 50 per cent of the knees injected, whereas 5.3 times 10-7 Micrococcus species caused only transient colonization of the joints. Methicillin, cephacetrile, and clindamycin administered before injection of the staphylococci gave protection. Stainless-steel particles in the knee did not increase susceptibility to infection from injected micrococci but did make established micrococcal infections more persistent. Knees containing steel debris failed to become infected during prolonged staphylococcal bacteremia. Injected autologous blood, either fresh or three days old, did not increase susceptibility to infection. Large doses of steroids, given systemically or intraarticularly, decreased resistance to micrococcal infection and increased the destruction caused by infection due to this organism.


Some physiological aspects of bone marrow pressure. An investigation was made into some of the hemodynamic properties of bone. The parameters studied were bone marrow and medullary venous pressures. The resting level and changes in the two pressures were measured under autoperfusion and constant-flow perfusion conditions. Changes were induced either by drugs or by alteration in blood flow. Marrow pressure was measured with a new tonometric technique. A specially constructed miniature pressure transducer was utilized as the tonometer. Trauma-induced hemorrhage of the marrow was avoided. The marrow pressure was measured across an intact endosteal membrane. The pressure of the normal bone marrow and the medullary venous pressure in the anesthetized dog were nearly equal, approximately twenty millimeters of mercury and about 17 per cent of systemic blood pressure. These values were found whether measured simultaneously or separately in comparable groups of dogs under a variety of conditions. Induced changes in the two pressures produced either by changes in perfusion rate or by drugs were also nearly equal and in the same direction.


The relative safety of polymethylmethacrylate. A controlled clinical study of randomly selected patients treated with Charnley and ring total hip replacements. Forty patients from a group of seventy elected to participate in a prospective randomized clinical study designed to evaluate the systemic effects of polymethylmethacrylate implanted with a Charnley total hip replacement, utilizing patients treated with a Ring total hip prosthesis ad controls. Intraoperative alterations in cardiovascular function were related to use of methylmethacrylate, but postoperative changes in pulmonary function, erythrocyte sedimentation rate, serum lactic acid dehydrogenase, serum glutamicoxalacetic transaminase, and alkaline phosphatase were not. Charnley total hip replacements were associated with more ectopic-bone formation twelve months postoperatively. The two "latent" infections recognized during the thirty-four months of the study were in hips without acrylic fixation.


Transplantation of preserved composite tendon allografts. Composite flexor-tendon grafts (both tendons with sheath and pulleys intact) were performed in adult chickens comparing fresh autografts with allografts stored in 1:5,000 merthiolate under refrigeration for two months. The gross and microscopic findings were assessed one week to one year after transfer. The allografts were somewhat slower to heal but otherwise differed little from the autografts, and were readily accepted eventually with only minor adhesions between the grafted tendon and the sheath.


Alterations of rabbit articular cartilage by intra-articular injections of glucocorticoids. The destruction of rabbit articular cartilage after intra-articular injections of hydrocortisone acetate was investigated using histological, biochemical, and tracer methods. Fissures and cysts increased in number as increasing amounts of hydrocortisone were given. A linear decrease of hexosamine to less than 50 per cent after twelve injections was accompanied by insigificant changes in deoxyribonucleic acid and hydroxyproline content. The synthesis of proteoglycans and proteins was reduced to one-third, while the production of collagen dropped to less than one-fifth. The changes in thymidine incorporation were not significant. Based on these data, a model indicating the sequence of events which leads to joint destruction after intra-articular injections of glucocorticoid is proposed.


Water content and binding in normal and osteoarthritic human cartilage. Water binding in normal and osteoarthritic cartilage slices from human femoral heads was studied using tritiated water to measure the water binding after short exposure, total water content, and avidity with which water is held. In osteoarthritic cartilage, compared with normal cartilage, there was no increase in water binding but water content increased by 9 per cent and the avidity with which the newly bound water was held also increased. These findings were almost exactly duplicated by partial extraction of the proteoglycan from normal tissues with 4 molar guanidinium hydrochloride.


The effect of operating-room environment on the infection rate after Charnley low-friction total hip replacement. After 300 Charnley low-friction arthroplasties done in two hospitals by two surgical teams using clean-air enclosures but no preoperative, operative, or postoperative antibiotics, there were three deep wound infections, two caused by Staphylococcus albus, coagulase negative, and one by Klebsiella. Two of these infections occurred in hips previously operated on and one was in a hip with no previous surgery. These findings suggest that two operating-room environments using the same basic principles without antibiotics can achieve a 1 per cent rate of deep wound infection with no early deep wound infections after total hip replacement, a rate comparable to that reported in other series in which antibiotics were used. However, this study did not answer the questions: Is the environment the primary reason for the low infection rate or is it the discipline required by the environment? Will the rate of late (four to five years) wound infection after operations done in a clean-air enclosure be lower than that after procedures done in a "normal" operating-room environment using preoperative, operative, and postoperative antibiotics?


Malignant hemangioendothelioma of bone.

Malignant hemangioendothelioma of bone. The clinical and histopathological characteristics are described in seven cases of malignant hemangioendothelioma of bone. This rare neoplasm affects all age groups and almost any bone. Metastases appear in the skeleton even before such lesions are seen on pulmonary roentgenograms. Prognosis is poor, with a five-year survival for only two of the six patients in this series. One patient is still alive and free of symptoms more than twelve years after diagnosis of widespread tumor destruction of the frontal skull bones, which was successfully controlled by radiation therapy. In addition, a recently diagnosed case of this tumor arising in a chronic non-fistulous osteomyelitic bone is described.


Factitious lymphedema of the hand.

Factitious lymphedema of the hand. Twenty-two patients with factitious lymphedema of the hand were reviewed. Thirteen were female, with a median age of sixteen years at the onset of symptoms. Of the nine males, the median age at onset of symptoms was thirty-two years. The dominant hand was affected more frequently than the non-dominant hand. Edema was usually caused by a tourniquet, irritation of the skin, or blows to the back of the hand. Neurosis, psychosis, or suicidal tendency was diagnosed in twelve of these patients. Characteristically, the edema was not particularly painful; it frequently ended proximally in a circumferential discolored constriction ring; and it occasionally displayed characteristic lymphangiographic findings of normal or dilated lymphatic channels with increased collateral circulation and multiple blowouts. Although ten of the patients received Workmen's Compensation benefits, the course and psychiatric diagnosis indicated that malingering for secondary financial gain was not a primary goal. Hospitalization is usually required to confirm the diagnosis. Psychotherapy is indicated once the diagnosis is made.


Synovectomy for rheumatoid arthritis of the knee. Synovectomy of the knee results in a satisfactory degree of pain relief in a patient with rheumatoid arthritis. Pain relief was equally good in our series in knees with clinically active synovitis, regardless of the amount of joint destruction, provided no gross malalignment, instability, or degeneration of joint surfaces was present. In 64 per cent of the knees there was some loss of range of motion; in 13 per cent, a gain in range of motion; and in 23 per cent, no change in the total range of motion. Only two knees in the entire series underwent fibrous ankylosis. The patients' estimates of their disease activity after surgery correlated very closely with the pain relief obtained. Most of those who estimated their activity to be very low had good pain relief, while those who had more active disease had less likelihood of a satisfactory result. Although long-term satisfactory results seemed to indicate that synovitis had been effectively suppressed by surgical synovectomy, the data in this series do not prove that synovectomy prevents destruction in the rheumatoid knee.


Forearm fractures. Early functional bracing - A preliminary report. Forty-five fractures of the forearm in forty-three patients were treated with a functional below-the-elbow brace applied three to forty-two days after injury, at a median time of fifteen days. The brace permitted flexion and extension of the wrist and elbow while limiting pronation and supination of the forearm during the fracture's reparative period. The results were rewarding, with only one non-union and minimum impairment of function in the overwhelming majority of cases.


Pins and plaster treatment of comminuted fractures of the distal end of the radius. The technique used in the treatment of seventy-five patients with severely comminuted, often intra-articular fractures near the wrist was as follows- Pins were inserted through the metacarpals and proximal part of the ulna, reduction was done, and the pins were incorporated in a cast from elbow to knuckles. This allowed movement of the fingers and the elbow. The main complications - loss of radial length and pintract infection - are avoided by attention to technical details. Of the patients with adequate follow-up, 86 per cent had good or excellent results.


Colles' fractures. Functional bracing in supination. The classic position of immobilization of Colles' fractures with the elbow in flexion, the forearm in pronation, and the wrist in volar flexion and ulnar deviation is probably the main reason for the common and rapid recurrence of the original deformity. Such a position places the brachioradialis muscle, a strong flexor of the elbow and the only muscle attached to the distal fracture fragment, in an ideal physiological position to exert a deforming force on the fracture fragments. Based on this assumption, further supported by electromyographic studies, a method of treatment was developed which calls for the initial immobilization of the arm in an above-the-elbow cast with the elbow in flexion, the forearm in supination, and the wrist inmoderate ulnar and volar flexion. This cast is changed a few days after application for an Orthoplast brace that permits motion of the elbow and volar flexion of the wrist while preventing pronation and supination of the forearm and dorsiflexion of the wrist. The proposed method did not prevent collapse of the fragments in all instances. However, the degree of collapse was minimum. The position of supination of the forearm and the freedom of motion of all joints seemed to reduce the swelling, stiffness, and incapacitation frequently found during active treatment of these fractures.


Experimental fixation of fractures of the femur with methylmethacrylate. Use of a simple method for injection of cooled methylmethacrylate into the distal part of the femur through the canal of a Küntscher rod to stabilize distal femoral-shaft fractures proved experimentally to greatly increase the torsional stability at the fracture site. This method may allow rapid ambulation of debilitated patients and prevent the complications of extended bed rest.


Fractures of the tibial shaft after skiing and other accidents. Fractures of the tibial shaft from skiing accidents were compared with fractures of other etiology. Skiing fractures more often affected the lower third of the tibia and they seldom were open. Many were comminuted or transverse but displacement tended to be slight. The incidence of complications was low and the periods of hospitalization, of healing, and of inability to work were shorter than for the other fractures. The highly favorable prognosis was evident whatever the method used for treatment. Methods of treatment of tibial fractures should not be compared on the basis of experience with skiing fractures, because of the exceedingly favorable prognosis of the latter.


Acute, complete acromioclavicular separation. Twenty-three patients with acute, complete acromioclavicular separation were studied prospectively. Twelve patients were treated non-operatively and eleven were treated operatively. A twelve-month minimum follow-up, using a point system to evaluate clinical results, demonstrated comparable results with and without surgery. Roentgenographic findings did not correlate with the clinical results. Minimum immobilization and early rehabilitation of the shoulder was the recommended treatment of choice.


Avascular necrosis of the proximal end of the dens. A complication of halo-pelvic distraction. Roentgenographic evidence of avascular necrosis of the upper part of the dens was observed in thirteen of ninety-four patients treated for tuberculous kyphosis or non-tuberculous scoliosis using halo-pelvic distraction. Four patients had some residual loss of motion. A significant relationship was demonstrated between avascular necrosis and the amount of distraction visible on roentgenograms.


Kinesiology after McKee-Farrar total hip replacement. A two-year follow-up of one hundred cases. Measurements of functional performance were made before and at six and twenty-four months after 100 McKee-Farrar total hip replacements in eighty-three patients. The measurements included: range of motion of the hip, hip-muscle torque, weight-bearing activity during standing, forces applied to canes or crutches, and multiple components of walking performance. In the absence of serious complications, most patients had improvement in most components of function, with greater gains occurring during the first six months and lesser but continued gains between the sixth and twenty-fourth postoperative months. Patients with serious operative complications, postoperative infections, or component loosening showed declines in almost every component of function tested.


Chondromalacia of the patella in Chinese. A postmortem study. Two hundred knees of 100 Chinese cadavera were studied with reference to factors which have been implicated in the etiology of chondromalacia of the patella. The findings were as follows: chondromalacia of the patella was less common in Chinese than in Europeans, did not always start in the medial patellar facet, and showed no relationship to the presence of abnormal menisci or to the anatomical characteristics of the medial femoral condyle which have been suggested as causative factors.


Syme's amputation. Follow-up study of weight-bearing in sixty-eight patients. Sixty-eight patients with Syme amputations resulting from industrial injuries were reviewed. The amount of end-weight-bearing was measured with a capacitance transducer. The majority of patients required fitting with a prosthesis designed to relieve end-weight-bearing. The old-fashioned prosthesis with a leather corset was the most effective in achieving reduction of end-weight-bearing.


Scoliosis in Marfan's syndrome. Its characteristics and results of treatment in thirty-five patients. Of sixty-four patients with Marfan's syndrome, thirty-five had scoliosis, which in 44 per cent began in the infantile and juvenile age periods. The curve patterns were double major right thoracic-left lumbar in 48 per cent and single right thoracic in 33 per cent. At the start of treatment the curves were severe (average, 72 degrees) and quite structural (average bending correction, 36 per cent). Seventy-four per cent of the thirty-five patients had back pain. Inadequately treated and untreated curves usually progressed to severe and painful deformity. Milwaukee brace treatment was generally unsuccessful because of the severity and rigidity of the curves or the poor maintenance of correction obtained. Spine fusion combined with Harrington rod instrumentation (fourteen patients) gave an average correction of 41 per cent with an average loss of 7 degrees (average follow-up, 2.3 years). Operative treatment of scoliosis is indicated in most patients with Marfan's syndrome.


Healing of electrosurgical and scalpel wounds in rabbits. Skin wounds in rabbits were tested after healing periods of up to three weeks to determine if scalpel wounds differed from those made electrosurgically. Tensile tests showed that the electrosurgical wounds were initially stronger, but at four days there was no difference. Thereafter the scalpel wounds were stronger and their healing progressed much faster. Histological preparations showed more extensive inflammation and necrosis in the electrosurgical wounds.


A characterization of polymethylmethacrylate bone cement. Polymethylmethacrylate cement is characterized in terms of chemical composition, handling characteristics, and physical properties; the dough time, setting time, handling time, and temperature rise were found to be most affected by environmental temperature and kneading of the dough mass. As the set material ages, the residual monomer content gradually decreases and the strength increases. A volume shrinkage of up to 5 per cent was observed. The porosity, which is increased by rapid mixing, may reach 10 per cent. The mechanical properties of the cement approximate those of polymethylmethacrylate denture base material. Some variability encountered in the handling qualities of the cement could be attributed only to the variability of different batches.


Biochemical confirmation of an experimental osteoarthritis model. Section of the medial collateral and both cruciate ligaments combined with resection of the medial meniscus in rabbit knees caused instability and during the ensuing six months these knees showed progressive histological changes similar to those of human osteoarthritis. Biochemical analysis of the cartilage from such knee joints demonstrated a decrease in proteoglycan, an increase in acid phosphatase, and increases in the rates of synthesis of protein and glycosaminoglycan. These findings, which are quite consistent with those in human osteoarthritis, suggest that this animal model may be of value in the study of the pathogenesis and treatment of human disease.


Acute neurological complications in the treatment of scoliosis. A report of the Scoliosis Research Society. A survey conducted by the Scoliosis Research Society found eighty-seven patients with acute neurological complications resulting from the treatment of scoliosis. The incidence of these complications was determined to be 0.72 per cent. Seventy-four major complications involving the spinal cord were reported, half of them complete paraplegia and half partial paraplegia. Thirty-six per cent recovered completely, 32 per cent had partial recovery, and 32 per cent had no return of function. Thirteen minor complications involving cranial and peripheral nerves were reported. Major complications occurred in forty-two cases of posterior spine fusion with Harrington instrumentation and in twenty cases of posterior spine fusion without instrumentation. Six patients became paraplegic following skeletal traction alone.


Traumatic dislocation of the knee joint. The results in thirty-three of fifty-three traumatic dislocations of the knee followed for more than one year confirmed our conclusion, made in 1971, that early repair of all torn ligaments gives the best results, and that the vascular status following this injury must be observed carefully since vascular repair or thrombectomy within eight hours of injury gives the best chance of preventing gangrene of the leg. The twenty associated fractures in these thirty-three patients were treated successfully with conventional methods, except for three displaced fractures of the medial tibial plateau in which closed reduction failed and internal fixation was required.


Exposure of an arginine-rich protein at surface of cells in S, G2, and M phases of the cell cycle. Phenylglyoxal (PG) is shown to be a cell surface probe specific for arginine moieties in protein: (1) It does not enter the cell as evidenced by lack of PG in the cytoplasm. (2) It does not cause excessive cell leakage as measured by release of 51Cr. (3) It reacts with positively-charged groups in proteins at the cell surface but not with those of phospholipids at the surface; since pronase removes PG from the surface, but phospholipase C does not. (4) Under the conditions used in these experiments, it reacts virtually exclusively with arginine moieties in protein (Freedman et al., '68; Takahashi, '68; Werber and Sokolovsky, '72). Synchronized cells were exposed to radioactive PG to assess quantity of arginine moieties in protein at the surface. There is a sharp decrease in arginine at the cell surface at entry into G1 phase from M and a 24-fold increase upon entry into S phase. There is a slight drop in exposed arginine in late S phase followed by an increase to 26 times the G1 level immediately prior to mitosis. Lactoperoxidase-catalyzed iodination of tyrosine moieties in protein at the surface of synchronized cells shows a very gradual increase in protein as the cells move through the cycle and increase in size. Since the increase in arginine moieties in protein at the surface does not reflect a similar increase in total protein at the surface, an arginine-rich protein appears to be exposed at the cell surface during the division-related phases of the cell cycle.


Conditionally lethal mutations in chinese hamster cells. Characterization of a cell line with a possible defect in the Krebs cycle. A variant Chinese hamster cell line has been isolated from a mutagenized population that has a markedly reduced ability to oxidize a variety of substrates via the Krebs cycle. The production of 14CO2 from 14C-labeled compounds was measured using pyruvate, acetate, beta-hydroxybutyrate, palmitate and glutamate, and in all cases it was neglibible in the mutant. In contrast to this, significant amounts of 14CO2 were produced from 14C-aspartate and 14C-succinate which suggest that some reactions of the Krebs cycle can take place and this conclusion is supported by tracer experiments with labeled compounds. The rate of respiration measured with a Clark oxygen electrode in the mutant was compared to several normal Chinese hamster cell lines and was found to be only 8%. Mitochondria appear to be present in normal numbers and with only minor differences in morphology. The measurement of difference spectra between oxidized and reduced states permits us to conclude that the cytochromes are all present and functional. These results lead us to believe that there may be a defect in the Krebs cycle between alpha-ketoglutarate and succinate. Alternatively a defect in a structural component of the mitochondria or in the electron-transport chain itself may be causing pleiotropic effects in the Krebs cycle and respiration.


Steroid hormone regulation of specific messenger RNA and protein synthesis in eucaryotic cells. Evidence is presented that the induction of specific proteins in the chick oviduct by the steroid hormones estrogen and progesterone, involves a primary effect at the level of gene transcription. The intracellular levels of mRNA's which code for the synthesis of the egg-white proteins, ovalbumin and avidin, have been quantitated in a heterologous protein synthesizing system. It is demonstrated that these levels are directly dependent upon the inducing steroid, estrogen or progesterone, respectively. Ovalbumin mRNA has been purified to apparent homogeneity. This ovalbumin mRNA was then used as a template for the synthesis of a complementary DNA copy catalyzed by the enzyme reverse transcriptase which was isolated from avian myeloblastosis virus. This radioactively labeled complementary DNA was used to demonstrate, by means of DNA excess hybridization, that the ovalbumin gene is represented only once in each haploid genome of the chick cell. Next the complementary DNA copy of the ovalbumin mRNA was used as a genetic probe to determine the precise number of sequences of ovalbumin mRNA present at any one time after the administration of estrogen. It was demonstrated that the unstimulated chick contained no sequences of ovalbumin mRNA. Within a very short period of time after estrogen is administered the ovalbumin sequences begin to appear and reach a steady state level of 140,000 molecules per tubular gland cell. It could also be calculated that each ovalbumin molecule is probably translated some 50,000 times during its life which explains why ovalbumin comprises some 60% of the total protein in the oviduct cell. Following withdrawal of the oviduct from estrogen treatment, ovalbumin mRNA sequences again drop to undetectable levels. However, following a single injection of estrogen to these withdrawn animals, new ovalbumin mRNA sequences could be detected within 30 minutes. These data suggest that estrogen controls the activity of the ovalbumin gene via a pure transcriptional control mechanism. It is also demonstrated that the efficiency of the complementary DNA as a means of quantitating specific mRNA sequences is some 1,000 times more sensitive than the best available in vitro translation system. Finally, the efficacy of four popular translation systems is compared. It is suggested that for initial studies involving hormonal control of mRNA levels, the translation system derived from wheat germ is the simplest and most sensitive.


The molecular genetics of mammalian glucuronidase. The genetic factors known to be involved in the final realization of beta-glucuronidase activity in mice are considered from the standpoint of structural genes determining the catalytic activity of enzyme molecules as well as the recognition features of enzyme molecules that identify them for subsequent processing by the cell; processing genes determining the cellular apparatus involved with the conjugation, intracellular localization and eventual degradation of enzyme molecules; regulatory genes determining rates of enzyme synthesis, especially in response to physiological signals such as hormones; and temporal genes determining the developmental programs for expression of these classes during growth and differentiation. The properties of genetic variants of beta-glucuronidase falling into each of these classes are described. When those results are considered in concert with the properties of genetic variants known for other mammalian enzymes several generalizations emerge. Structural genes of enzymes are not usually linked to the processing genes determining the post-assembly events in the life of that enzyme. In contrast, all of the regulatory and temporal gene sites so far identified are in close proximity to the structural genes they modulate. Regulatory and temporal sites appear to act in a cis fashion to control the amount of enzyme synthesized from the adjacent structural allele on the same chromosome.


The genetics and developmental regulation of L-glycerol 3-phosphate dehydrogenase. In addition to reviewing the genetic regulation of L-glycerol 3-phosphate dehydrogenase during development in the mouse, new evidence is presented that the electrophoretic properties of L-glucerol 3-phosphate dehydrogenase in Mus castaneus are determined by an allele (d) at the Gdc-1 locus. Accordingly there are three alleles at the Gdc-1 locus; the b allele in C57BL/6J mice differs from the d allele in electrophoretic properties and the c allele in BALB/cJ mice differs from the d allele with respect to both heat denaturation and electrophoretic properties. Identical segregation patterns of the L-glycerol 3-phosphate dehydrogenase phenotypes in liver, kidney, and skeletal muscle from offspring of an F2 generation produced from parents with the c/d genotype suggest that the Gdc-1 locus is the major structural locus for L-glycerol 3-phosphate dehydrogenase in these tissues. Heart muscle was pooled from mice of the F2 generation with either c/c or d/d genotypes at the Gdc-1 locus as determined by analysis of liver L-glycerol 3-phosphate dehydrogenase. The chromatographic properties of L-glycerol 3-phosphate dehydrogenase from the heart muscle was determined on DEAE-cellulose ion exchange columns. The elusion profile of the L-glycerol 3-phosphate dehydrogenase indicates that the Gdc-1 locus is also the major structural locus in heart muscle.


Gene activation during spermatogenesis.

Gene activation during spermatogenesis. Cell differentiation during spermatogenesis in the rat has been analyzed in terms of the formation of specific "marker" enzymes. Hyaluronidase and other acrosomal enzymes are formed in spermatids according to a highly predictable time schedule which may be termed a "molecular biological clock". The acrosomal enzymes beta-galactosidase and N-acetyl-beta-glucosaminidase exist as isoenzyme forms distinct from enzymes with similar substrate specificities in the lysosomes of precursor cells. Differentiation of spermatids thus involves the loss of gene expression for lysosomal enzymes and the activation of genes for acrosomal isoenzymes. Spermatogenesis is characterized by the sequential loss of expression of many genes, as evidenced by the loss of beta-glucuronidase in the differentiation of spermatogonia to spermatocytes, and the loss of uridine diphosphatase activity in the differentiation of spermatocytes to spermatids. The apparent absence of ornithine decarboxylase activity from spermatids suggests a dependence of these cells upon Sertoli cells for the provision of putrescine and/or spermidine. Such biochemical cooperativity among germinal cells may be necessary as the genes of spermatids are repressed and late spermatids become metabolically inactive. Spermatogenesis is also characterized by changes in the cellular content and rates of synthesis and phosphorylation of specific acidic chromatin proteins. It is hypothesized that these proteins may participate in the activation or repression of genes during spermatogenesis.


Homocystinuria due to cystathionine synthase deficiency. Studies of nitrogen balance and sulfur excretion. Apparent nitrogen balances and urinary sulfur excretions were determined for normal subjects, seven cystathionine synthase-deficient patients, and a single cystathioninuric patient on semisynthetic diets containing low-adequate amounts of methionine and very low amounts of methionine and very low amounts (12 mg daily, or less) of cystine. The amounts of supplemental cystine required to prevent abnormally high nitrogen or sulfur losses were determined. The five cystathionine synthase-deficient patients who had low residual activities of this enzyme detected in fibroblast and/or liver extracts did not lose more nitrogen or sulfur on diets virtually devoid of cystine than did the normal subjects. These results suggest that the widely expressed opinion that cystine is an essential amino acid for cystathionine syntase-deficient patients requires modification. Residual enzyme activity of only a few percent of normal may obviate such a cystine requirement. These results are compatible with, and lend support to, the working hypothesis which states that the pyridoxine response in cystathionine synthase-deficient patients is mediated by an increase in the residual activity of the affected enzyme.


The effect of estrogen on the lipoprotein lipase activity of rat adipose tissue. The effect of 17beta-estradiol or progesterone administration on adipose tissue lipoprotein lipase activity was studied in male and ovariectomized female rats. Lipoprotein lipase activity was measured in acetone-ether-extracted preparations of adipose tissue with doubly labeled (14C-fatty acid, 3H-glyceryl) chylomicron triglyceride as substrate. Administration of 17beta-estradiol to male rats lowered adipose tissue lipoprotein lipase activity from 8.22 plus or minus 1.8 U/g (1 U = 1 mumol triglyceride hydrolyzed per h) to 4.96 plus or minus 0.5 U/g in the treated group. Ovariectomy increased adipose tissue lipoprotein lipase activity from 10.4 plus or minus 1.8 U/g in controls to 22.7 plus or minus 4.3 U/g. 17beta-Estradiol administration to ovariectomized rats cuased a marked fall in adipose tissue lipoprotein lipase activity: 17beta-estradiol (2.5 mug/day) lowered the enzyme activity to 9.00 plus or minus 1.2 U/g, whereas 25 mug/day further decreased lipoprotein lipase activity to 3.2 plus or minus 0.6 U/g. Blood triglyceride levels increased from 0.8 plus or minus 0.05 mumol/ml in ovariectomized rats to 1.4 plus or minus 0.09 mumol/ml in 25 mug/day 17beta-estradiol-treated rats. Progesterone administration did not affect adipose tissue lipoprotein lipase activity in either male or ovariectomized rats. Heart and lung lipoprotein lipase activity was unaffected by hormone treatment. We suggest that the rise in blood triglyceride concentrations, which accompanies high palsma estrogen levels, could be due to the marked inhibition of adipose tissue lipoprotein lipase activity.


Evidence that histidine is an essential amino acid in normal and chronically uremic man. The requirement for dietary histidine was investigated in four normal and three chronically uremic men. Subjects lived in a metabolic unit where they were fed three isonitrogenous diets in the following order: a 40-g protein diet (28 plus or minus SD 8 days), a semi-synthetic amino acid diet deficient in histidine (35 plus or minus 2 days), and an amino acid diet which contained histidine (31 plus or minus 5 days). With ingestion of the histidine-deficient diet, nitrogen balance gradually became negative, and serum albumin decreased in six subjects. Plasma histidine fell by 82 plus or minus 6 per cent; muscle histidine decreased by 62 plus or minus 19 per cent; the hematocrit fell by 25 plus or minus 9 per cent; and serum iron rose. Subjects felt unwell, and in five cases a skin lesion consisting of fine scales, dry skin, and mild erythema developed. After administration of the histidine-repletion diet, nitrogen balance became positive in  six subjects; serum albumin increased in five cases; plasma and muscle histidine rose; serum iron fell abruptly; a reticulocytosis ensued; and the hematocrit rose. The clinical symptoms and skin lesions disappeared. These observations indicate that histidine is an essential amino acid in normal and chronically uremic man. The absence of dietary histidine is associated with failure of normal erythropoiesis.


The influence of hypertonic mannitol on regional myocardial blood flow during acute and chronic myocardial ischemia in anesthetized and awake intact dogs. The influence of hypertonic mannitol on regional myocardial blood flow and ventricular performance was studied during acute myocardial ischemia in awake, unsedated and in anesthesized dogs and after myocardial infarction in awake unsedated dogs. Regional myocardial blood flow was measured with radioactive microspheres. Generalized increases in regional myocardial blood flow occurred after mannitol in all of the different animal models studied. The increases in coronary blood flow after mannitol were just as impressive in the nonischemic regions as in the ischemic portion of the left ventricle in all of the different models that were examined in this study. Improvement in regional myocardial blood flow to the ischemic area of the left ventricle after mannitol was associated with a reduction in ST segment elevation during acute myocardial ischemia in anesthetized dogs. The increases in regional myocardial flow after mannitol were also associated with increases in contractility, but the increases in flow appeared to be more impressive than the changes in contractility. The data obtained demonstrate that mannitol increases regional coronary blood flow to both ischemic and nonischemic myocardium in both anesthetized and awake, unsedated, intact dogs with acute and chronic myocardial ischemia and that mannitol reduces ST segment elevation during acute myocardial ischemia in anesthetized dogs. Thus the results suggest that under these circumstances the increases in regional myocardial blood flow after mannitol are of physiological importance in reducing the extent of myocardial injury. Since coronary blood flow increased to nonischemic regions the increases in regional myocardial flow demonstrated in this study after mannitol cannot be entirely explained by the mechanism of reduction in ischemic cell swelling.


Isolation and characterization of two hydroxyproline-containing glycoproteins from normal animal lung lavage and lamellar bodies. Two glycopeptides, present in particulate material obtained by pulmonary lavage from normal rabbits, were isolated and characterized. The same two glycopeptides were present in preparations of lamellar bodies from rabbit lung. The estimated molecular weights of the two glycopeptides by sodium dodecyl sulfate-acrylamide gel electrophoresis were found to be 62,000 and 36,000 and both were found to contain hydroxyproline and relatively high amount of glycine (11 and 15 per cent, respectively). Carbohydrate analysis of the two glycopeptides demonstrated the presence of glucosamine, sialic acid, mannose, Fucose, and galactose. Similar glycopeptides of the same molecular weights, and amino acid and carbohydrate compositions have been found in layage mateial isolated from lungs of patients with alveolar proteinosis. The data indicate that thses two collagen-like glycopeptides are major intra-alveolar proteins in many mammals, including humans.


The pathogenesis of arthritis associated with acute hepatitis-B surface antigen-positive hepatitis. Complement activation and characterization of circulating immune complexes. Circulating immune complexes were identified in cryoproteins isolated from serial samples of serum from six patients with acute viral hepatitis with and without arthritic symptoms. Cryoprecipitates were analyzed for the presence of hepatitis-B surface antigen (HBsAg) and hepatitis-B surface antibody (anti-HBs) by hemagglutination inhibition and hemagglutination. Complement components were detected by counter electrophoresis, and immunoglobulins were detected by gel diffusion. HBsAg, IgG, and IgM were identified in cryoprecipitates from all hepatitis patients, but were higher in concentration in patients with arthritis. Only cryoprecipitates from hepatitis patients with arthritis contained IgA and complement components C3, C4, and C5 as well as IgG and IgM, which disappear with resolution of the arthritis. The subtypes of IgG in these cryoprecipitates were predominantly the complement-fixing IgG1 and IgG3, HBsAg and anti-HBs were concentrated several-fold in the cryoprecipitates when compared to the serum concentration. Sequential studies in two patients demonstrated that the initial appearance of anti-HBs in the cryoprotein complex was associated with the detection in the complex of IgM suggesting a primary immune response to HBsAg. The C3 activator fragment (C3A) of the properdin complex was found in fresh serum obtained from three hepatitis patients with arthritis and not in uncomplicated hepatitis. The cryoprecipitable immune complexes from patients with arthritis converted C3PA in fresh normal sera to C3A in vitro whereas cryoprotein isolated from patients with uncomplicated hepatitis had no such effect. Thus, the transient appearance of circulating complement-fixing immune complexes in patients with the arthritis of acute hepatitis is associated with activation of both classical and alternate complement pathways and suggests that they play an important role in the pathogenesis of these serum sickness-like extrahepatic symptoms.


Effects of the 15-methyl analogs of prostaglandins E2 and F2alpha on the pulmonary circulation in the intact dog. The effects of the 15-methul analogs of prostaglandins E2 (PGE2) and F2alpha (PGF2alpha) on the pulmonary circulation were studied in the intact dog under conditions of controlled blood flow. Infusions of either analog into the lobar artery increased lobar arterial pressure by more than 100 per cent. The rise in lobar arterial pressure was accompanied by a rise in lobar venous pressure and in pressure gradient from lobar artery to small vein but no change in pressure in the left atrium. The methyl analogs were about 10 times more potent than PGE2 and PGF2alpha in elevating pulmonary vascular resistance in the dog. The effects of the analogs on the pulmonary vascular bed were similar in experiments in which the lung was perfused with dextran or with blood. Both analogs contracted isolated helical segments of canine intrapulmonary artery and vein in a dose-related manner. In other experiments the effects of passive increases in venous pressure produced by distension of a balloon catheter in the lobar vein were contrasted with the action of the analogs on the pulmonary vascular bed. Balloon distension increased pressure in the lobar artery and small vein but had no effect on pressure in the left atrium. However, in contrast to the increase in gradient with the analogs, balloon distension decreased the pressure gradient from lobar artery to small vein. Results of the present study indicate that the prostaglandin analogs increase pulmonary vascular resistance by actively contricting pulmonary veins and vessels upstream to small veins, presumed to be small arteries. It is concluded that the analogs are potent pressor substances in the pulmonary circulation.


H2O2 release from human granulocytes during phagocytosis. I. Documentation, quantitation, and some regulating factors. The extinction of fluorescence of scopoletin during its oxidation by horseradish peroxidase (HPO) provides a highly sensitive and specific assay for small quantities of peroxide in solution. With this assay, the release of free H2O2 into the extracellular medium by phagocytizing human granulocytes has been documented and quantitated, and some of the regulating factors have been determined. Under basal conditions granulocytes released less than 0.01 nmol/ml of H2O2 (2.5 X 10-6 polymorphonuclear leukocytes/ml). Upon the addition of phagocyte particles (latex, opsonized yeast, or staphylococci), an abrupt increase in extracellular peroxide concentration was observed (greater than 50-fold above basal levels) after latencies as short as 10 s. Release reflected increased intracellular H2O2 production during phagocytosis in that it paralleled the respiratory burst and was absent when phagocytosis was prevented or when cells from patients with chronic granulomatous disease were utilized. Evidence that scpoletin oxidation occurred predominantly in the extracellular medium was obtained by demonstrating a marked inhibition when HPO was omitted from the reaction mixture or when exogenous catalase was added. Similarly, it was found that exogenous serum also inhibited scopoletin oxidation, apparently because of the presence of competing hydrogen donors. H2O2 formation  and release were observed at rates which closely paralleled those of phagocytosis. With O2 consumption as an approximate index of H2O2 formation, the fractions released during maximal rates of particle uptake were calculated as follows: for latex, 15.7%; for staphylococci, 10.3%; and for yeast, 4.9%. It is postulated that release is due to diffusion of free H2O2 from an expanded intracellular pool of this substance that develops during phagocytosis. This poos represents tha net of increased synthesis versus catabolism by various enxymatic pathways for H2O2 disposal within the cells. The close relationship between rates of H2O2 formation and rates of phagocytosis by human granulocytes suggests a role for specialized areas of the cell membrane, involved in particle ingestion, in the trigger mechanism for H2O2 synthesis. The consequences of H2O2 release to other cells or organisms in the immediate environment of phagocytizing granulocytes remain to be determined.


Interaction of leukocyte chemotactic factors with the cell surface. I. Chemotactic factor-induced changes in human granulocyte surface charge. The negative surface charge of human granulocytes was diminished after incubation with the chemotactic factors C5a, dialyzable transfer factor, and the enzymes kallikrein and plasminogen activator. No such change was observed after incubation with human IgG, albumin, horeseradish peroxidase, or a mixture of prekallikrein and plaminogen proactivator. Hydrocortisone inhibited the effect of C5a upon granulocyte surface charge and inhibited its chemotactic activity, suggesting that steroids act at the cell surface. The chemotactic inhibitors cholchicine and cytochalsin B had no effect upon granulocyte surface charge, consistent with their presumed effect upon microtubules and microfilaments, respectively. The data suggest that the decrease in cell surface charge may be a preerequiste for normal cell movement.


Human adipocyte cholesterol. Concentration, localization, synthesis, and turnover. By analysis of 124 specimens in 16 different patients, isolated human adipocyte cholesterol concentration is highly correlated with fat cell size but not with plasma cholesterol concentration. Less than 6 percent of total cholesterol is esterified; after subcellular fractionation, 88 percent of the cholesterol is recovered in the triglyceride-rich supernatant oil. This latter finding supports the observation that fat cell cholesterol is determined by triglyceride content, and hence by fat cell size. After intravenous administrtion of radioactive cholesterol, the sum of a three-exponential equation was fit simultaneously to both the plasma and adipocyte specific activity time curves in six patients. In five of the six, a slowly turning over pool (pool 3) closely fit the adipocyte data. Two model structures, mammillary and catenary, were fitted to the data. There was no synthesis in pool 3 using a mammillary model but a mean 5.3 percent of the total body production rate was found in compartment 3 if a catenary model was assumed. Although a catenary model is biologically unlikely, it could not be excluded. Obesity is associated with an increased cholesterol synthetic rate equal to 20 mg/day for each kilogram of body fat. To test (by an independent method) if this synthesis might be occurring in adipose tissue, human fat cells were obtained under a wide variety of dietary conditions and incubated in vitro with radioactive glucose or acetate. Incorportation of these precursors into sterol could account for no more than 1 mg cholesterol synthesis/kg fat per day. These in vitro data taken together with the in vivo mammillary compartmental analysis data are compatible with the possiblity that the excess cholesterol synthesis of obesity occurs in pool 1, most likely from hepatic or intestinal sites.


A micropuncture study of renal salt and water retention in chronic bile duct obstruction. The mechanism of sodium retention by the kidney in rats with ligation of the common bile duct was studied with micropuncture techniques. 10-14 days after bile duct ligation, rats showed positive sodium balance and ascites formation. Measurements of renal blood flow and glomerular filtration rate yielded values that were not different from those in normal control animals. Likewise, single nephron filtration rte of surface nephrons was the same in the experimental rats as in the controls. Sodium reabsorption, however, was markedly increased in the proximal convoluted tubule, as well as in segments beyond the proximal convolutions. Single nephron filtration fraction, calculated from measurements of efferent arteriolar and arterial hematocrits, was significantly elevated in the cortical nephrons, even though whole kidney filtration fraction was the same as in normal rats. The calculated protein concentration of cortical peritubular blood was higher in the bile duct-ligated rats than in the normal controls. The observations are consistent with the view that sodium retention is the result of enhanced reabsorption primarily by cortical nephrons. The enhanced reabsorption can be accounted for by relative cortical ischemia due to efferent arteriolar vasoconstriction with the consequent elevation of peritubular colloid oncotic pressure.


A sensitive technique by differential scanning for detecting haemoglobins of fetal origin. A simple modification is made to the ultraviolet scanning technique for the detection of the tryptophan notch which is characteristic of haemoglobins of fetal origin. This involves a differential scan using Hb A solution in the reference compartment. This technique has all the advantages of the standard scan method but extends the scope of it by being much more sensitive and decisive.


The flora of renal haemodialysis shunt sites. During investigations of the microbial flora of the skin over haemodialysis shunt sites it has not proved possible to predict clinical infection by a preceding colonization of the shunt site with a pathogenic organism. The normal non-pathogenic flora of the sites is not specifically related to the flora of other sites on the body though Staphylococcus aureus on a shunt site appeared to be acquired principally from the nose when the shunt was in the arm or from the perineum when the shunt was in the leg. Cimino shunt sites had a greater density of organisms than did Scribner shunt sites; this may be related to the disinfection procedures.


Intact and fragmented intracellular immunoglobulin in a case of non-secretory myeloma. A patient is described with myeloma without paraproteinaemia or Bence Jones proteinuria in whom the tumour cells have been shown to contain monoclonal immunoglobulin. The use of immunoelectrophoresis and polyacrylamide gel electrophoresis has established, apparently for the first time, that the immunoglobulin components are present in the form of intact molecules and free light chains.


Indexing and filing of pathological illustrations. An inexpensive feature card retrieval system has been combined with the Systematised Nomenclature of Pathology (SNOP) to provide simple but efficient means of indexing and filing 2 in. x 2 in. transparencies within a department of pathology. Using this system 2400 transparencies and the associated index cards can be conveniently stored in one drawer of a standard filing cabinet.


Haematological changes in active chronic hepatitis with reference to the role of the spleen. The haematological role of the spleen has been investigated in a series of 22 patients with active chronic hepatitis. Severe pancytopenia occurred in one patient after three years of steroid therapy and this episode was associated with an increase in spleen size and a high splenic index of red cell destruction. Although the spleen was usually enlarged in the remainder of treated and untreated patients no others showed increased splenic haemolysis. The red cell survival was slightly reduced in most patients but splenic pooling of red cells and expansion of the plasma volume did not significantly reduce the haematocrit level. No consistent haematological differences were detected between the untreated and the treated patients.


Segmental glomerulonephritis.

Segmental glomerulonephritis. The renal biopsy findings in 40 patients with segmental glomerulonephritis are reported. The term is used to describe a condition in which one or more segments of the glomerular tuft is involved by disease when other segments appear unaffected on light microscopy. The word 'focal' is not used as it may be taken to imply that the changes affect some glomeruli but not others and the evidence for this is not convincing. Segmental glomerulonephritis was a relatively common finding in cases of proteinuria with or without the nephrotic syndrome. The severity of the glomerular changes did not correlate with the ultimate prognosis. On the other hand the tubular and interstitial changes, as assessed by a grading procedure and by point counting, were significantly less severe in those patients who showed clinical recovery than in those who did not.


An ultrastructural study of the urinary bladder in children correlated with histological, bacteriological, and clinical findings. Bladder biopsies from six boys without a history of urinary tract infection were taken during hypospadias repair operations and examined by electron microscopy. The ultrastructure of the epithelium, which was presumed to represent the normal, is described in detail and compared with the reported findings in other mammalian species. The appearances are generally similar although in our material the luminal membrane of the superficial cells was thicker than that surrounding the other epithelial cells but not asymmetrical. We observed membrane-coating granules which have not been reported before in bladder epithelium. The appearances of the normal bladder are compared to those seen in 24 children with urinary infection. In the presence of acute infection in large heterogeneous secondary lysosomes which are present in the intermediate and superficial cells of the normal bladder are reduced in number and the amount of rough endoplasmic reticulum is increased.


Herniation of mucosal epithelium into the submucosa in chronic ulcerative colitis. Herniation of the glandular epithelium into the submucosa has been observed in 11 out of 27 cases of chronic ulcerative colitis. Glandular herniation was associated with thickening of the muscularis mucosae, with interruption of the muscularis mucosae by lymphoid follicles, and, in five of the 11 cases, with significant crowding of the glands of the mucosa. This study strongly suggests that sustained contraction of the muscularis mucosae, which has been shown by others to be a major feature of chronic ulcerative colitis, is the prime factor in the formation of downgrowths or herniations of the glandular epithelium into the submucosa. Comparison of the cases in which cancer developed with those where there was glandular herniation led to the conclusion that they are independent associations of chronic ulcerative colitis, and that glandular herniation plays no part in the development of dysplasia or cancer.


Detection of tetracycline resistance in Streptococcus pneumoniae. Bacteriological details are given of a patient with chronic purulent bronchitis, who was being followed up during a survey of relapse in chronic bronchitis. A strain of Streptococcus pneumoniae, serotype 10, was isolated from the sputum over a period of six months, followed by a type 47A strain and later a type 28 strain. The patient was receiving prophylactic treatment with tetracycline throughout. The type 10 strain was sensitive to tetracycline in vitro by both the disc diffusion and doubling dilution sensitivity tests and mice infected with this strain were protected by tetracycline. In contrast, both the type 47A and type 28 strains were sensitive by the disc diffusion technique, but showed a low degree of tetracycline resistance by the doubling dilution method; mice infected with both these strains were not protected by tetracycline.


The efficacy of long-term oral anticoagulant therapy and its laboratory assessment. The activated partial thromboplastin time is compared with the corresponding prothrombin ratio in 6378 samples of platelet-poor plasma from 446 patients treated for a total of more than 4500 patient/months with oral anticoagulatnts. A relative decrease in the activated partial thromboplastin time following deep vein thrombosis is described, which tends to become less obvious during the first year of treatment and is greater in older patients. Although this relative decrease is also found in patients treated after cerebrovascular accidents, it is not found in patients treated after myocardial infarction or in patients with mitral valve disease treated prophylactically with long-term oral anticoagulants. It is though possible that these changes following deep vein thrombosis might be useful in helping to determine the duration of oral anticoagulant treatment.


Cardiovascular drug interactions.

Cardiovascular drug interactions. Drug interactions may be responsible for certain changes in therapeutic response and toxicity of cardiac drugs. Interactions occur at the sites of drug absorption and elimination as well as at the receptor sites in the pacemaker cells, specialized conducting tissue, and myocardium. Studies of the kinetics of cardiac drugs are being applied clinically in an effort to reduce the danger of adverse drug interactions in heart patients.


Rheoencephalographic and other studies of betahistine in humans: I. The cerebral and peripheral circulatory effects of single doses in normal subjects. Two groups of six young normal male subjects were studied by an improved form of rheoencephalography (intracranial rheoncephalography), by photoplethysmography, and by impedance plethysmography to investigate the possible effects of single doses of betahistine hydrochloride (SERC) and placebo on the normal human cranial, cerebral, scalp, and calf circulations. Two subjects participated in both groups. The results of the two studies were similar and were combined for this presentation. One subject reported slight transient faintness and visual blurring after 20 mg of the drug. No other adverse or side effects were encountered. The results show that the drug causes definite, strong, reproducible, and dose-related responses in the studied circulations, typically those of decreases in their waveform amplitudes and pulse propagation times. Betahistine hydrochloride thus acts as a potent cerebral and peripheral microcirculatory vasodilator in normal humans when given orally. Split-dose studies indicate that responses followed each dose and that the second response was superimposed on the first. The value of using betahistine hydrochloride in overlapping doses is suggested. Atypical cranial, cerebral, scalp, and calf amplitude increases were found in some high-dosage (16-mg) trials. These increases may indicate that betahistine hydrochloride can also act as an arterial vasodilator in sufficiently high dosage, but they equally may indicate that sufficiently profound microcirculatory vasodilatation can elicit secondary arterial circulatory increases.


Cytoarchitecture of the optic tectum in the nurse shark. The cytoarchitecture of the optic tectum of the nurse shark is described and related to the arrangements of afferents from retina, telencephalon and contralateral tectum. Its lamination is not pronounced when compared to tecta of most other non-mammalian species but more comparable to those of mammals. The absence of highly differentiated cells such as pyramidal and true horizontal cells is perhaps correlated with the poor differentiation in general, including the apparent partial overlap of inputs. Some neurons near the midline were found to possess dendrites extending into the contralateral tectum.


Quantitative relationships between nerve and satellite cells in spinal ganglia: an electron microscopical study. II. Reptiles. In the spinal ganglia of two species of reptiles (gecko and lizard) the volume of the perikaryal satellite cell sheath was found directly proportional both to the volume and surface area of the related neuronal body. This result agrees with that obtained in a previous research on two species of mammals (cat and rabbit). A quantitative balance between neuronal bodies and their associated glial tissue therefore exists also in the spinal ganglia of zoological species phylogenetically quite distant from mammals. The quantitative relationship between glial and nerve tissue was found to be lower in the gecko and lizard than in the cat and rabbit. This difference could have a phylogenetic significance, and/or it could be explained by the lower metabolic rate in the nervous system of the poikilotherms in respect to mammals.


A brief self-assessing depression scale. The Wang-Self-Assessing Depression Scale (SADS) was devised to provide a brief self-rating form for measuring depressive symptomatology. The present study compares the SADS with the Zung Self-Rating Depression Scale (SDS) to assess reliability and relative ease of completion. Ninety-three ratings on each scale were obtained from a subject group that included normal volunteers and patients with differing degrees of depression. The paired t-test showed no significant difference between mean SDS scores and mean SADS scores for normal volunteers or subjects rated at any of the four depression levels. Positive correlation was demonstrated between tsds scores and SADS scores for depressed and normal subjects. The period of time required to complete the Wang SADS was found to be significantly shorter than for the Zung SDS, while the number of errors and requests for additional assistance were significantly lower. It was felt that these differences would constitute an advantage in the clinical use of the Wang SADS for diagnosing, evaluating, and monitoring the progress of depressed inpatients and outpatients.


Plasma levels of a new pelletized form of amitriptyline for maintenance therapy. Plasma levels of amitriptyline and nortriptyline were measured in volunteers after one week of daily dosage with either 25 mg t.i.d. of standard amitriptyline tablets or a single 75-mg capsule of a new pelletized form of amitriptyline. Concentrations of both compounds were essentially equal at the end of one week's dosage with either dosage form but were somewhat higher throughout the dosing time interval for the pelletized drug.


Autoradiographic investigations of glial proliferation in the brain of adult mice. II. Cycle time and mode of proliferation of neuroglia and endothelial cells. The cycle time of the proliferating glial cells outside the subependymal layer of the lateral ventricle as well as that of endothelial cells was studied autoradiographically in the brains of adult and untreated mice. To determine the mean cycle time two independent methods were used. A mean cycle time of about 20 hours was obtained for glial and endothelial cells from the decrease of the mean grain number/nucleus as a function of time after tritiated thymidine (3H-TdR) injection. Another group of experiments utilized the "method of labeled S phases". With this method the passage of labeled cells through successive S phases is observed. Passing through S phase following 3H-TdR injection the 3H-labeled cells are double labeled by an additional 14C-TdR injection. This method again resulted in a cycle time of 20 hours for glial and endothelial cells. From the present work and a former study (Korr et al., '73) the following cell cycle parameters were derived: Cycle time 20 hours; S phase 9.4 hours; G2 less than three hours; (G2+M) five hours; G1 five hours. The growth fraction of glial cells related to all glial cells is only 0.004. Furthermore, the present experiments show that in the case of glial cells 17% of the daughter cells after mitosis become pyknotic and are eliminated from the glial cell population. Apart from this cell loss, after mitosis about one-fourth of the daughter cells do not enter the next S phase. These cells leave the growth fraction and are replaced by a corresponding number of non-proliferating glial cells. There is a relatively extensive permanent exchange of cells between the growth fraction and non-growth fraction of glial cell.


Kinetics of salicylates in blood and joint fluid. Samples of blood and joint fluid from 30 patients who had taken acetylsalicylic acid were examined for concentrations of total salicylates. The data were arranged in groups according to diagnosis of the joint disease. Analysis of the data did not show significant differences in the kinetics of total salicylates into blood. In groups, the time to first appearance of 0.3 mg/liter averaged 6 minutes, and levels "close to maximum" concentration averaged about 23.0 mg/liter. Time to first appearance of 0.3 mg/liter of total salicylates in joint lfuid ranged from 10 to 31 minutes. The average level of maximum concentration of total salicylates in joint fluid was 14.8 mg/liter. Transport of total salicylates from blood to joint fluid showed a pattern consistent with the type of joint disease. Support was found for the hypothesis that diffusion was the major factor in the process of movement of total salicylates from blood to joint fluid. The histopathologic changes in different types of synovitis influenced the biophysical properties of the inner part of the joint capsule.


The inferior olivary nucleus of the opossum (Didelphis marsupialis virginiana), its organization and connections. Although the inferior olivary nucleus of the opossum is small, sections stained either for Nissl substance, normal axons or cholinesterase activity reveal distinct medial, dorsal and principal nuclei. The medial nucleus contains three major subdivisions (labelled a, b, c after Bowman and Sladek, '73) and a group of neurons which is comparable to the cap of Kooy. In contrast to the cat and monkey, the major portion of the "medial" nucleus (subgroup a) lies lateral to the principal nucleus in rostral sections. The dorsal nucleus can also be subdivided, as can the principal nucleus which contains distinct dorsal and ventral lamellae. A small area is identified which based on position and connections may conform to the dorsal medial cell group. The experimental portion of the study provides evidence for an olivary projection from the motor-sensory cortex and a massive input from the midbrain (red nucleus, pretectum, midbrain tegmentum). In addition, the opossum inferior olive receives fibers from the deep cerebellar nuclei (cerebellar feedback loops), the spinal cord and the dorsal column nuclei. Of particular interest is the finding that fibers from the nucleus cuneatus and nucleus gracilis have distinctly different olivary targets and that those from the nucleus gracilis, but not the cuneate nucleus, overlap (in part, at least) with the direct spinal fibers. Other examples of overlapping fields of terminal degeneration are present and are discussed. In general our results reveal that although certain relationships between the nuclear divisions are different, the opossum olive conforms well to that of placental mammals and provides a basic mammalian model for future experimental electron microscopic and physiological studies.


Clinical effects of pentazocine in hospitalized medical patients. Of 17,068 hospitalized medical patients monitored in a drug surveillance program, 616 (3.6 per cent) received pentazocine hydrochloride orally and 816 (4.8 per cent) received pentazocine lactate parenterally during one or more admissions. Unsatisfactory analgesic efficacy as judged by each patient's physician was nearly twice as frequent among recipients of pentazocine lactate as among recipients of parenteral morphine or meperidine. Adverse reactions were attributed to pentazocine hydrochloride in 18 recipients (2.9 per cent) and to pentazocine lactate in 37 recipients (4.5 per cent). Neuropsychiatric effects were the most common; they included hallucinations, bizarre feelings, disorientation, and agitation. These effects were dose dependent and frequently serious in nature. Minor gastro-intestinal effects were the second most common adverse reactions.


Cell death in the development of the lateral motor column of the chick embryo. Cell counts were made in the lumbar lateral motor column (l.m.c.) of chick embryos of 5.5, 6, 7, 8, 9, 12, 18 days of incubation and five days posthatching (n equal 68). Only nuclei with nucleoli were counted and corrections were made for double counting (Abercrombie, '46). The population attains a peak value of over 20,000 cells (corrected figure: over 17,000) at 5.5-6.5 days equal stages 28 and 29 (Hamburger and Hamilton, '51). The l.m.c. loses between 7,000 and 8,000 cells between days 6.5 and 9.5, (between stages 29 and 36). In other words, 60% of the population survive. A plateau of approximately 12,300 cells (corrected figure: 10,300) is maintained through five days posthatching. Massive cell degeneration was observed in 7- and 8-day embryos. Counts of distinctly pyknotic cells indicate that at least 5-6% of the total population is in the process of degeneration at any particular time. This figure is probably an underestimation; hence it is virtually certain that the depletion of the l.m.c. is due entirely to cell death. Arguments are preue to the failure of their axons to survive in a competition process at the periphery. Observations of the time pattern of muscle differentiation and their neurotization in the leg further endorse this hypothesis. However, it is not clear whether the axons compete for contact sites on muscle fibers or for a "trophic" agent.


Interspecies study on the effect of perphenazine on milk yield and composition. The effects on lactation of perphenazine, a tranquillizer which increases the level of prolactin in the blood, have been studied in cows, goats, guinea-pigs and rabbits. The drug caused a significant depression of milk yield in cows, goats and rabbits, but was without effect on the milk yield of guinea-pigs. It seems probable that the inhibitory effects on lactation were due to the actions of the drug on appetite and behaviour. It is concluded that perphenazine, because of its other actions, is not a suitable agent for studying the physiological effects of increased levels of endogenous prolactin.


Influence of sympathomimetic drugs on the motility of bovine teat muscles. Spontaneous rhythmical contractions of the teats of living cows were recorded by a plethysmographical method. These contractions were inhibited by isoproterenol, adrenaline, isoxsuprine and phentolamine, but were activated by phenylephrine. Noradrenaline induced activation which was often followed by inhibition. These results obtained in vivo were in agreement with responses obtained in vitro. Apparently beta sympathomimetic effects are evoked easily under in vivo conditions.


Preparation and properties of alpha S-casein from buffalo's milk. Buffalo alpha S-casein (alpha-sb) was found to be heterogenous; on starch gel-urea electrophoresis 4 electrophoretic components were detected. The alpha-sb-fraction was isolated by urea and alcohol fractionation. A preparation containing only the 2 main components was obtained by chromatography on DEAE-cellulose. Its amino acid pattern was different from that of cow alpha s1-casein. The fraction was also free of carbohydrate, cystine and cysteine. Buffalo alpha-sb-casein was less stable than cow alpha s-casein in solutions containing Ca, and the maximum stability of the alpha s-/k-casein complex occurred at a lower Ca concentration.


Autoxidation in milk rich in linoleic acid. I. An objective method of measuring autoxidation and evaluating antioxidants. Autoxidation was studied in milk obtained from cows fed on formaldehyde-treated casein-safflower oil supplement. The fat of this milk contained high levels of linoleic acid. A rapid disappearance of dissolved oxygen (DO), measured by using a oxygen electrode, from milk samples stored at 0 degrees C in tubes without head-space, coincided with the development of oxidized flavours detected organoleptically. A correlation coefficient of 0.9 with 69 degrees of freedom (P smaller than 0.001) was obtained  between the amount of DO disappearing and the taste panel scores for oxidized flavours. Butylated hydroxyanisole, sesamol, nordihydroguaiaretic acid, ethoxyquin, or butylated hydroxyanisole with propylgallate or tocopherols, when added in emulsified form to the milk at the rate of 10-15 mg/l milk checked the development of oxidized flavours and the rapid disappearance of DO. Other antioxidants tested were either ineffective or imparted off-flavours to milk. Samples of mare's milk neither developed oxidized flavours nor showed rapid disappearance of DO over a test period of 8 d. The oxygen electrode provides a convenient and sensitive method for studying autoxidation and evaluating antioxidants in milk.


Nasal mucosal hyperpermeability to macromolecules in atopic rhinitis and extrinsic asthma. This study was designed to test the hypothesis that patients with atopic rhinitis and extrinsic asthma have a nasal mucous membrane defect that allows inhaled macromolecules access to immunocompetent cells. Three groups were studied: normal subjects, patients with extrinsic asthma, and patients with atopic rhinitis. Albumin 125I(20 muc) was applied to the nasal mucosa and venous blood samples were drawn at set intervals up to 4 hours. Thirty-two minutes after administration, a significantly greater percentage of the dose was found in the plasma of patients with atopic rhinitis than in that of normal subjects (p smaller than 0.001). Transport of intact albumin across the nasal mucosa was demonstrated by dialysis, gel filtration, and immunopercipitation experiments in 3/9 normal subjects, 1/9 patients with asthma, and 9/10 patients with rhinitis (p smaller than 0.02). These studies suggest that large, potentially antigenic molecules pass more readily across the nasal mucous membrane of patients with allergic rhinitis than that of normal subjects. No increase in nasal transport was seen in patients with extrinsic asthma. It has not been determined whether this defect is a cause or an effect of atopic rhinitis.


Growth of asthmatic children during treatment with alternate-day steriods. The effects of specific doses of alternate-day treatment with prednisone on linear growth were evaluated in children with severe asthma. It was found that even the control patients who did not receive steroid therapy had heights that were significantly lower than those of normal children of the same age and sex. The average severity of growth suppression in children who received alternate-day or intermittent treatment with steriods did not differ from that of asthmatic control patients. However, evaluation of individual patterns of growth during the follow-up period revealed that children who received small doses of alternate-day treatment (mean dose of prednisone, 9 mg.  q.o.d.; range, 2.5 to 14 mg.) had acceleration of growth, whereas children who received larger treatment doses (mean dose of prednisone, 30 mg. q.o.d.; range, 18 to 58 mg.) had further suppression of growth during the period of study. Additionally, patients who had previously been treated with daily corticosteroids failed to demonstrate "catch-up" growth after introduction of an alternate-day program (mean dose of prednisone, 17 mg. q.o.d.).


Measurement of the prevalence of respiratory allergies by interview questionnaire. A recent publication of the National Center for Health Statistics estimated the prevalence of asthma in 1970 to be 30.2 per 1,000 noninstitutionalized population, and of hay fever (without asthma) as 54.2 per 1,000. These and other data on chronic respiratory conditions were obtained by means of a household survey employing lay interviewers and a standardized questionnaire. There may be a gap between the intended meaning of the key question-did the individual have a chronic condition during the past 12 months-and the respondent's understanding of it. As a consequence, some of those who have been asymptomatic in the recent past may have excluded themselves. Even among people with the chronic condition who have seen a physician about it during the prior year, significant numbers fail to report it to the interviewer, according to validation studies. A method is suggested for making rough adjustments for these undercounts. It results in raising the estimated prevalence to 43.0 for asthma and 71.1 for hay fever. These revisions are dependent on necessarily subjective assumptions about the magnitude of the biases involved.


The dietetic paraprofessional and the external degree. Report on work in progress. The "University without Walls", i.e., the external degree earned by work off-campus, is a relatively new but increasingly important development in higher education. At Pennsylvania State University, such a program leading to an associate degree is being developed for the dietetic technician. Through correspondence study for credit and other avenues, dietetic paraprofessionals can prepare themselves for upgrading to supervisory roles calling for academic preparation. The need is particularly apparent in view of recently issued requirements that supervisory foodservice personnel in extended care facilities have 90 clock hours of instruction. The projected curriculum will permit the earning of credits by correspondence; through courses taught by multi-media; by transferring courses from other institutions; or by examination. Elements of the curriculum planning are detailed.


Comparison of Type A and nutrient standard menus for school lunch. I. Development of the nutrient standard method (NSM). Methodology was developed for planning school lunch menus which meet a specific nutrient standard. The method considered nine indicator nutrients plus calories and percentage of calories coming from fat. The nutrient composition of approximately 625 school lunch menu items were calculated using Agriculture Handbook No. 8 data. All nutrient data were converted to bead units which were summed on an abacus until the meal requirements were met. Preliminary testing of the method showed it to be usable by school lunch menu planners and provided menus meeting certain minimal nutrient constraints. After the test, suggested modifications were incorporated into the method to enhance its use. The method is described and a sample week's given.