The effects of diethyl ether, enflurane, and isoflurane at the neuromuscular junction.

The actions of diethyl ether, enflurane, and isoflurane at the neuromuscular junction were examined in isolated guinea pig lumbrical muscles. These anesthetics depressed the ability of carbachol to depolarize the endplate region; this depression of depolarization did not show competitive kinetics. None of the anesthetics altered the affinity of the acetylcholine receptor for d-tubocurarine, i.e., the dissociation constant of d-tubocurarine was unchanged. Since diethyl ether, enflurane, and isoflurane produced no observable alteration of the receptor, the antagonism of the drug-induced depolarization of the neuromuscular junction appears to be exerted at a stage subsequent to reaction with the receptor. (Key words: Anesthetics, volatile, diethyl ethers; Anesthetics, volatile, euflurane; Anesthetics, volatile, isoflurane; Neuromuscular relaxants, d-tubocurarine; Neuromuscular junction.).


Surgical management of high aortoillac occlusion.

Seventeen patients with high aortoiliac occlusion, 15 chronic and two acute, were evaluated at the Walter Reed Army Medical Center during the period 1964-1973. Fifteen patients with chronic occlusion and one patient with an acute occlusion were operated upon without an operative death. The operative technique outlined emphasizes temporay interruption of renal blood flow during the initial aortic thromboendarterctomy phase of the procedure. In those patients in whom renal artery control was secured, no instance of postoperative renal insufficiency was noted. Symptomatic improvement occurred in all patients. Ultimate follow-up results will be dependent on the amount of associated vascular disease.


Anastomotic aneurysms.

This report is concerned with an analysis of 16 anastomotic aneurysms which developed in 12 patients after arterial surgery in which dacron sutures were used for all graft anastomoses. Knitted dacron grafts had been used in all but two of the original surgical procedures; a woven dacron graft had been used in one procedure and a saphenous vein graft in the other. The majority of anastomotic aneurysms occurred in the femoral triangle at the site of an end-to-side anastomosis. Five anastomotic aneurysms were caused by infection, and a variety of etiologic factors were responsible for the 11 non-infected aneurysms. Fourteen anastomotic aneurysms in 10 patients were treated surgically. A good result had been obtained in six patients; one patient required amputation of an extremity and three patients died postoperatively as a result of cardiac disease or pulmonary embolization. This report also contains a discussion of the factors affecting the occurrence of anastomotic aneurysms and a description of specific operative measures designed to decrease the incidence of these aneurysms.


Filtration characteristics of three new in-line blood transfusion filters.

Three types of microemboli filters were examined with the scanning electron microscope. The single layer 40 mu pore filter traps aggregates in the pores. The aggregates are extruded and pass on with the blood and can be demonstrated in a Dacron wool filter downstream. The polyurethane foam filter is an in depth multiple pore filter of variable pore size. This filter allows many aggregates to escape through the pores in the beginning of a transfusion and these aggregates were seen in a Dacron wool filter downstream. As aggregates become trapped, the pore sizes decrease, filtration improves, and flow through the filter becomes reduced. The Dacron wool filter absorbs the particulate material to the fibers of wool. This filter has a large capacity and was not observed to leak aggregates even when subjected to high filtration pressures.


Development and treatment of post-traumatic pulmonary platelet trapping.

In anesthetized dogs soft tissue trauma resulting in hypovolemia was associated with pulmonary trapping of 51Cr tagged platelets. The efficacy of various forms of therapy aimed at preventing pulmonary trapping of platelets following trauma was evaluated. Lung biopsies were assayed before trauma and at hourly intervals for accumulation of autologous 51Cr tagged platelets reinfused 24 hours prior to the experiment. The study showed that pulmonary platelet trapping secondary to soft tissue trauma could be prevented by treatment with Dextran 40, Dextran 70 and methyl-prednisolone administered one hour after trauma and by pretreatment with aspirin.


The value of laparoscopy in acute pelvic pain.

Laparoscopy was performed in 223 patients with acute pelvic pain but without a definite diagnosis. The clinically suspected diagnosis was confirmed by laparoscopy in only 57 patients (25%). Laparotomy was thus avoided in 145 patients (65%). The endoscopic findings in the three clinical entities included here are presented: tubal pregnancy, acute appendicitis or torsion of adnexal mass. This study emphasizes the poor correlation between the clinical diagnosis based on history, pelvic examination and physical signs, and the final laparoscopic findings. The value of laparoscopy in evaluation of acute pelvic disease is stressed.


Hemodynamics of the interposition mesocaval shunt.

Eighty interposition mesocaval shunts, using a knitted Dacron large diameter prosthesis, have been performed during the past five and one-half years. Patients were evaluated from the standpoint of protection from recurrent esophageal hemorrhage, shunt patency, encephalopathy and cumulative survival analysis. In a selected group of patients, hemodynamic measurements were also obtained in the pre, intra, and postoperative periods. These included measurements of wedged hepatic vein pressure, superior mesentric venous blood flow, and residual superior mesenteric, hepatic sinusoidal and inferior vena cava pressures following the shunt procedure. Additionally, direct shunt flow measurements utilizing a square wave of electromagnetic flowmeter were also performed. Results indicate that the shunt patency is 95%; adequate decompression of the portal system was accomplished; recurrent variceal hemorrhage did not occur if the shunt remained patent; the incidence of encephalopathy was low (11%); and the operative mortality for the entire series was 9%. Continued perfusion of the liver was documented in 44% of patients and appears to be a function of the residual total portal resistance largely controlled by inferior vena caval pressure at the level of graft replacement. Life survivhat the interposition mesocaval shunt appears to be an effective technique for the control of variceal hemorrhage, has important hemodynamic advantages and can be applied to most patients for the control of variceal hemorrhage due to portal hypertension.


Surgical management of abdominal aortic aneurysms: factors influencing mortality and morbidity--a 20-year experience.

Abdominal aortic aneurysmectomy is being performed with progressively lower operative mortality and morbidity. Three hundred thirty seven patients have had elective aneurysm repair since 1954. Factors affecting mortality and morbidity in the last 108 cases are analyzed. Seventy-four per cent of patients had pre-existing disease, either cardiac, pulmonary, renal, cerebrovascular, diabetes mellitus, or hypertension. Six patients died following operation, a mortality rate of 5.5%. One died of pulmonary and 5 of cardiac causes. No patient died of renal failure or required dialysis. A signficant feature of management is the regimen of fluid therapy using dextrose in lactated Ringer's solution during and after operation to minimize  hypotensive and renal complications. No patient developed a wound infection, graft infection, wound dehiscence, stroke, or intestinal ischemia. Serious postoperative complications were largely cardiac or pulmonary. Despite recent liberalization of indications for operation, comparative figures show continued reduction in operative mortality from 17% during 1954-1961, or 7.4% during 1962-1967, to 5.5% in the 1968-1974 era. This declining mortality is related to earlier diagnosis using non-invasive methods (sonogram), simplified operative techniques, improvement in fluid management, innovations in cardiopulmonary therapy, and recognition and proper handling of unusual manifestations of aortic aneurysms.


New technique for construction of coposite Dacron vein grafts for femoro-distal popliteal bypass in the severely ischemic leg.

Vascular surgeons are in agreement that autogenous saphenous veins are best suited for bypasses from the common femoral artery to the distal popliteal artery in the management of femoropopliteal occlusive disease associated with the severely ischemic foot. Such a graft should be of adequate size (more than 3 mm in diameter) throughout its length for a successful outcome. In some patients the vein is of good size for 15 or 20 cm then branches into several small veins. Reports by most surgeons are unfavorable concerning the use of prostheses and bovine heterografts for anastomosis to the distal popliteal artery or to one of its branches. Our experience with composite dacron vein graft bypasses employing a fluted end-to-end anastomosis had been unfavorable and was similar to the  experience of Dale (1962). In July 1973 we were forced to improvise the technique of end-to-side anastomosis joining the end of a dacron prosthesis to the side of the vein graft for a femorodistal popliteal bypass. During the ensuing 15 months we have carried out this composite graft only when the greater saphenous vein was not of adequate size throughout. In 17 limbs the composite graft was placed between the common femoral artery and the distal popliteal artery and on 6 occasions to the posterior tibial and peroneal arteries. Nineteen limbs exhibited either gangrene, impending gangrene, ischemic ulceration or severe rest pain. In four extremities intermittent claudication of a progressive and disabling degree was the indication for operation. Eleven of the 22 patients were diabetic. Run-off beyond the popliteal artery was poor in 16 of the 23 limbs and inflow was subnormal in three patients. During the followup period, 10 grafts have occluded, one day to 6 1/2 months postoperatively, two due to inflow deficiency, 5 due to poor outflow, one to an error in technique, and two occluded without known cause. Two patients came to major amputation following closure of their grafts, 3 and 7 months postoperatively. Results with the composite graft are compared with the bovine heterograft and the homologous vein graft.


The use of Mersilene mesh in repair of abdominal wall hernias: a clinical and experimental study.

Use of synthetic materials in herniplasty has been a controversial issue. In order to determine the influence of Mersilene mesh on the strength of healing abdominal wounds and its effectiveness in repair of hernia, experimental and clinical studies were undertaken. Experimental study included 175 male rats divided into three groups subjected to either: 1) an incision made only through the skin and closed with 3-0 silk sutures; 2) a 2.5 cm midline incision through the musculature and peritoneum closed with 2-0 Mersilene suture; or 3) the same procedure as group 2 with the addition of a Mersilene mesh onlay graft. Bursting strength of abdominal wounds as determined in all groups at intervals. Wounds of the group treated with the mesh exhibited significantly greater (P less than 0.01) bursting strength. Clinical trial consisted of 100 consecutive adult patients in which an onlay graft of Mersilene mesh was used in the hernioplasty. Mesh was used as an adjunct in patients with: 1) large ventral hernias; 2) direct hernias resulting from severely attenuated transversialis fascia; 3) indirect hernias associated with a large internal ring and a weak posterior inguinal wall; or 4) combined direct and indirect hernias. All were followed for a minimum of one year to determine the incidence of complication and rate of recurrence. This study suggests that: 1) Mersilene mesh increases the strength of healing abdominal wounds in rats; and 2) repair of large hernias with Mersilene mesh results in an acceptable morbidity and a lowered rate of recurrence.


Recurrence after preperitoneal herniorrhaphy in the adult.

The recurrence rate was determined for 1,186 men aged 18 to 96 years (average, 56 years) operated on for inguinal herniation between Jan 1, 1968 and June 30, 1974. (Mean follow-up was three years five months). Fourteen hundred twenty defects were repaired through a modified preperitoneal approach. Eight patients (0.6%), average age 76 years, dide postoperatively. One hundred seven patients (9.0%), average age 65 years, have died since. Seventeen percent had bilateral herniation repaired simultaneously. Ninety-four (6.6%) have had recurrence. Indirect inguinal decects predominated (58%), a recurrence rate of 3.7%. Five hundred twenty-three primary direct defects were repaired; 8.2% recurred. There were 237 repairs for recurrence in 216 patients (average age, 59 years), a recurrence rate of 9.7%.


[Stroma formation in epithelial malignant tumors].

The mechanism of stroma-formation and rearrangement of the stroma was studied on 221 cases of cancer of the lung, stomach and mammary gland using histological, histochemical and electron-microscopy methods of investigation.  It was established that multiplying epithelial elements of teh tumour induced proliferation of histiogenic and vascular fibroblasts, activating them.  Active fibroblasts play a double role: they either produce fibres and interstitial matter of the tumorous stroma, or participate in desintegration of preceding and newly formed collagenous structures.  The processes of stroma-formation comply with the conventional schemes of normal fibrillogenesis and rearrangement of the interstitial tissue.  It was shown that fibroblasts of the tumour stroma was capable of phagocytosis of a mature collagen.  It is supposed that in the course of the tumorous growth correlation between the parenchyma and stroma is maintained, the leading role being played by the epithelium.


[The objectives and prospects of quantitative histocytochemistry].

The article is of a surveying character and is dedicated to objectives of quantititative histo mammary gland).  Problems connected with standardization of preparation of the material and carrying out of histochemical reactions, a complex of histochemical criteria and principles of their selection, are considered.  A variant of mathematical treatment of material by a computer, including analysis of distribution and correlation characteristic, is suggested.  The complex functional-morphological method (marker) suggested, with its quantitative assessment, is, in the opinion of the author, a qualitatively new basis for mathematical descriptions of organs and systems and their modelling and opens up new perspectives both in physiology and pathology.


Polypoid change of the glomerular basement membrane.

A peculiar and previously undescribed alteration in the glomerular basement membrane, which we designate as polypoid change, was found in a 4-year-old child with mongolism (trisomy 21). Light and electron microscopical studies were performed. Although the pathogenesis and significance of this lesion remain uncertain, we have compared the lesion with known processes that affect the glomerular basement membrane.


Changes in the electrocardiogram, vectorcardiogram and ballistocardiogram after hemodialysis in chronic uremia.

In 12 cases with uremia, the electrical activity and form and amplitude of Bcg (displacement) were investigated before and after hemodialysis. The uremia was secondary to chronic glomerulonephritis (9 cases), familial nephritis (1 case), polycystic kidney disease (1 case) and chronic pyelonephritis (1 case). During the study digitalis was not given to the patients. The ECG and Bcg abnormalities were irreversible in most cases. In 9 cases the VCG showed hypertrophy of the left ventricle. Considering these facts it is assumed that myocardial hypertrophy might play an important part in the development of irreversible pathological repolarization and hemodynamic troubles in chronic uremia.


Changes in aortic glycosaminoglycans and lipoprotein lipase activity in rats with age and atheroma.

The changes in the individual glycosaminoglycans of the aorta and in lipoprotein lipase activity of the aorta, liver and heart have been studied at various stages in the development of mild atheroma in the rat. Three responses were seen: (a) Hyaluronic acid initially decreased, then increased; (b) Heparan sulphate and chondroitin sulphates A and C initially increased, then decreased. (c) Chondroitin sulphate-B and heparin increased with progressing lipid infiltration and decreased markedly only in the later stages. Ageing changes were also investigated in the rat aorta: total cholesterol, phospholipids and triglycerides increased progressively from weaning to 9 months of age. Hyaluronic acid decreased after weaning, reached a minimum at 6 months and then increased thereafter. Heparan sulphate and chondroitin sulphate-C reached a maximum at 6 months and then decreased thereafter. Chondroitin sulphates A and B showed a similar but less marked pattern of change with age. Heparin progressively increased with age. Aortic lipoprotein lipase activity increased in the early stages of atheroma and then decreased as the lipid infiltration became more severe. The ageing study showed that enzyme activity was quite high at weaning. decreased considerably at 3 months, but thereafter fell only slightly.


Distribution of acidic glycosaminoglycans in the intima, media and adventitia of bovine aorta and their anticoagulant properties.

1. The constituents of acidic glycosaminoglycans in the tunica intima, media and adventitia were studied by means of an enzymatic assay with chondroitinases and by electrophoretic characterization. 2. The content of the acidic glycosaminoglycans was higher in the intima than in the adventitia based on defatted dry tissue weight. The gel filtration pattern showed that, with chondroitinase-AC, the proportion of the intact glycosaminoglycans is greater in the adventitia than in the intima. 3. Electrophoresis before and after digestion with chondroitinases indicated that the three layers contain chondroitin-4- and -6-sulfates, dermatan sulfate, heparan sulfates and hyaluronic acid. In the case of the adventitia, bands migrating like the heparin standard were also detected. 4. Paper chromatographic separation of the unsaturated disaccharides after digestion with chondroitinases revealed that the proportion of chondroitin-6-sulfate is predominant in the intima, whereas the proportion of heparan sulfates, hyaluronic acid and dermatan sulfate is one and one-half-2 times higher in the adventitia than in the intima. 5. Anticoagulant activity, measured by thrombelastography, of the acidic glycosaminoglycans in the three layers clearly indicated that the glycosaminoglycans in the adventitia possess the highest potency among the three layers. It was considered that the higher activity of the glycosaminoglycans in the adventitia is due to the relatively greater proportion of heparan sulfates and dermatan sulfate.


Metabolism of glycosaminoglycans in atheromatous rats. Enzymes concerned with synthesis, degradation and sulphation of glycosaminoglycans.

Some important enzymes concerned with the biosynthesis of the precursors of glycosaminoglycans (gg), degradation of gg and biological sulphation have been studied in rats fed an atherogenic diet. L-Glutamine-D-fructose-6-phosphate amino-transferase and glucosamine-6-phosphate-N-acetylase--2 enzymes concerned with the biosynthesis of hexosamine precursors  of gg--decreased in the liver in rats fed the atherogenic diet. UDPG pyrophosphorylase, UDPG dehydrogenase and UDPG glucuronic acid-5'-epimerase, which are concerned with the biosynthesis of the uronic precursors of gg, also decreased in the liver in the diet-fed rats. The activities of some of the enzymes concerned with degradation of gg-hyaluronidase, beta-glucuronidase beta-hexosaminidase, cathepsin and aryl sulphatase--increased both in the liver and aorta. The hepatic concentration of PAPS significantly decreased in the diet-fed rats. The sulphate-activating system, which includes ATP sulphurylase, APS kinase and sulphotransferase, also decreased. Thus the overall picture is one of decreased synthesis of gg and their increased degradation in the atheromatous rats.


Previous reproductive history in mothers presenting with spontaneous abortions.

Information on the previous reproductive history of 3467 mothers presenting with a spontaneous abortion is compared with that of a control series presenting with a livebirth. In 1384 abortions the chromosome constitution of the fetal products was determined. The mothers who had delivered a chromosomally normal abortion more often had a history of previous repeated abortions, while those of chromosomally abnormal abortions tended to have fewer repeated abortions but an excess of previous viable children with Down's syndrome. Data are also presented on 23 mothers with two spontaneous abortions of known karyotype; these, together with similar data obtained elsewhere, made it possible to estimate an overall rate of recurrence of chromosome defects in conceptions of the same parents, and the probability that affected fetuses will survive to a viable stage in pregnancy. The clinical and genetic implications of the findings are briefly discussed.


Some sources of stress found in mothers of spina bifida children.

The scores on a malaise inventory for 51 mothers of spina bifida children born between 1956 and 1962 in South Wales revealed significantly higher scores when the child was incontinent, had a severe locomotor disability, had an IQ less than 80, and was attending a special school rather than a normal school. No difference in scores was found between the sexes or for size of family, and only a slight difference occurred between the malaise scores of mothers of mildly and moderately handicapped children.


The Fowler-Weir approach to appendicectomy.

The medical approach of Fowler-Weir enables rapid opening of the abdominal cavity and provides good access to the appendix. Although there is less muscle trauma the wound seems as painful postoperatively as with the classic muscle-splitting approach. However, morbidity is less as shown by a decrease in the wound and deep infection rate compared with that after appendicectomy by the McBurney approach.


Atropine, sodium cromoglycate, and thymoxamine in PGF2 alpha-induced bronchoconstriction in extrinsic asthma.

In six patients with extrinsic bronchial asthma the inhalation of prostaglandin (PG) F2 alpha in a small dosage produced significant bronchoconstriction, whereas PGE2 produced bronchodilatation. In these patients cholinergic blockade with atropine partially inhibited the PGF2 alpha-induced bronchoconstriction, but the alpha-receptor-blocking drug thymoxamine and sodium cromoglycate did not. These results suggest that the effect of PGF2 alpha is mediated through cholinergic receptors in the airways, and this effect is grossly exaggerated in asthma. The failure to inhibit PGF2 alpha-induced bronchoconstriction with sodium cromoglycate and the observation of an inhibitory effect of sodium cromoglycate in both allergic and exercise asthma suggest that locally formed PGF2 alpha may not be the main factor in the pathogenesis of bronchial asthma.


Progress in understanding Huntington's chorea.

The present paper analyses the new data accumulated since 1972 concerning the etiology and pathogenesis of Huntington's chorea. Particular attention is paid to the respective roles of the dopaminergic and GABA-ergic systems.


Adverse reactions to trimethoprim-sulfamethoxazole, with particular reference to long-term therapy.

Since trimethoprim-sulfamethoxazole (TMP-SMX) was first marked occasional reviews have surveyed the pattern of adverse reactions. Skin rashes characteristic of sulfonamide sensitivity have predominated, with relatively few of a serious exfoliative nature. Hematologic adverse reactions recorded follow the pattern known to occur with sulfonamides, with a few cases related to the action of trimethoprim on human folate metabolism. Such an effect is more likely to occur when the patient's folate status is already jeopardized; it is rare in relation to the widespread use of the drug combination. Long-term administration does not per se seem to represent an additional hazard provided the dose is the correct one and the hematologic monitoring of the patient is performed regularly. The suggestion that TMP-SMX has a toxic effect on the kidney has not been substantiated. An estimated 250 million "standard treatment courses" have been given in the first 6 years of marketing and, overall, the picture of adverse reactions corresponds with that expected from a sulfonamide of relatively low toxicity.


A Mg2+- and Ca2+-stimulated adenosine triphosphatase at the outer surface of Ehrlich ascites tumor cells.

A Mg2+- and Ca2+-stimulated adenosine triphosphatase (ATPase) at the outer surface of intact Ehrlich ascites tumor cells is described. A surface-bound adenosine triphosphate (ATP)-splitting activity at a lower rate was also demonstrated in the absence of Ca2+ but with Mg2+, Na+, and K+ present in the isotonic medium. Hence, when part of the Mg2+ was exchanged for Ca2+, a marked increase of the ATP-splitting activity was observed. The stimulatory effect of Ca2+ was seen only if both Na+ and K+ were present in the isotonic incubation medium. Thus, the enzyme activity was Mg2+- and Ca2+-dependent. Ca2+, together with the monovalent cations was inhibitory compared with Mg2+ under similar conditions. The apparent Km for ATP for the Mg2+-stimulated ATPase is 0.05 mM, while that of the Mg2+- and Ca2+-stimulated enzyme is 0.10 mM. The Vmax of the former is 0.8 mu-mole per 100 mg Schneider protein per 30 sec compared with 1.92 mu-moles per 100 mg Schneider protein per 30 sec for the latter. The calculated Km for the Mg2+- and Ca2+-stimulated ATPase after subtraction of the Mg2+-stimulated part is 0.22 mM. Ethacrynic acid and N-ethylmaleimide both inhibited the Mg2+- and Ca2+-stimulated ATPase by about 10 percent, while the ouabain inhibition was 15 percent. Cytochalasin B did not influence the enzyme activity, whereas La3+ had a slight stimulatory effect.


Bioassay of alkyl halides and nucleotide base analogs by pulmonary tumor response in strain A mice.

The production of lung adenomas in strain A mice following multiple injections of 17 alkyl halides and of 3 base analogs was investigated. A slight but significant increase in the average number of lung tumors per mouse was noted following the administration of methyl iodide, n- and i-propyl iodide, sec- and tert-butyl chloride, i-, sec-, and tert-butyl bromide, and n- and sec-butyl iodide. The administration of comparable doses of ethyl bromide, ethyl iodide, n-butyl chloride, benzyl chloride, and 1-chloromethylnaphthalene to mice resulted in no significant increase in the frequency of lung tumors over that seen in vehicle-treated control mice. n-Butyl bromide and tert-butyl iodide similarly appeared to have no significant effect on the lung tumor frequency, but these compounds were too toxic to be tested at the high dosages used with the other alkyl halides. 5-Iodo-, 5-bromo-, and 5-fluorodeoxyuridine also appeared to have no significant effect on the lung tumor frequency. These results indicate that a high proportion of low-molecular-weight alkyl halides may be weakly carcinogenic and provide evidence supporting an electrophilic hypothesis of carcinogenesis.


The effect of flavonoids on aerobic glycolysis and growth of tumor cells.

Certain bioflavonoids inhibit the glycolysis of variety of tumor cells by interfering with the generation of adenosine diphosphate and inorganic phosphate which are required for glycolysis. Tetra- and pentahydroxy flavones with hydroxyl groups as 3, 3', 4', 5, and 7 (e.g., quercetin) are the most active. They inhibit the activity of isolated Na+-K+-adenosinetriphosphatase of the plasma membrane and of mitochondrial adenosinetriphosphatase, but under appropriate conditions do not interfere with the ion transport increase the the translocation efficiency of the ion pump. It was shown that in several tumor cells loosely coupled ion pumps are responsible for the high rate of aerobic glycolysis, the effect of quercetin on the growth of several cell lines was examined. Since bicarbonate and serum albumin were found to counteract the effect of quercetin, the cells were grown in tissue cultures at low concentrations of these compounds. Pronounced inhibition of growth was observed at 5 to 20 mug of quercetin per ml of growth medium.


A model of cell cycle control: effects of thymidine on synchronous cell cultures.

Further evidence is presented in support of a model for growth control in which commitment for cell division is determined by an event in the preceding cell cycle. A study was made of conditions affecting synchronous growth following treatment of murine mastocytoma cells with excess thymidine at different phases of the cell cycle. Cells were synchronized by a physical procedure involving velocity sedimentation in a zonal rotor. Pulse treatment of such cultures with thymidine at times corresponding to the S, G2, and M periods had no effect on further growth. However, addition at G1, although having no immediate effect, arrested cell growth in the next cell cycle. This temporal effect may account for the decay of synchrony observed during double thymidine blockade or thymidine-FUdR blockade. When the time interval between two such blocks was 7 hr or less, P815Y cells were arrested after one synchronous division. At this critical time a majority of cells were at, or near, G1. It is suggested that thymidine exerts a hitherto unrecognized effect at the G1 interval.


Evidence for a new mineralocorticoid in patients with low-renin essential hypertension.

Patients with low-renin essential hypertension have certain features consistent with excessive mineralocorticoid activity. Because known mineralocorticoids are normal in the majority of low-renin essential hypertension patients, an unknown mineralocorticoid was sought in the urine of such patients. Urine extracts from patients with low-renin essential hypertension were assayed for mineralocorticoid activity in adrenalectomized rats and found to contain more such activity than could be accounted for by the known mineralocorticoids in the extracts. The factor responsible for the unexplained mineralocorticoid activity was purfied and then identified by mass spectral analysis as 16beta-hydroxydehydroepiandrosterone (16beta-OH-DHEA). Synthetic 16beta-OH-DHEA was found to have a mineralocorticoid potency one-fortieth that of aldosterone in the rat bioassay. The mineralocorticoid effects of both the urine extracts and the synthetic steroid were blocked in the rat by spironolactone, a mineralocorticoid antagonist. A specific assay for 16beta-OH-DHEA was developed, and its level in the urine was found to be elevated in patients with low-renin essential hypertension.


Splanchnic blood flow in essential hypertension and in hypertensive patients with renal artery stenosis.

Splanchnic blood flow (SBF) was measured simultaneously with cardiac output (dye dilution) and intrarterial blood pressure by constant infusion of indocyanine green in 11 control subjects, 13 patients with essential hypertension (EH) and seven hypertensive patients with renal artery stenosis (RAS). The cardiac index (CI) was lower (P less than 0.05) in patients with EH (3.17 plus or minus 0.07 L/min/m-2) (mean plus or minus SEM) than in control subjects (3.43 plus or minus 0.09). Associated with the lower CI was a significantly (P less than 0.01) lower SBF (0.797 plus or minus 0.02 L/min/m-2 vs 0.889 plus or minus 0.04). Patients with RAS presented with higher (P less than 0.01 vs EH, nonsignificant vs control subjects) cardiac index (3.66 plus or minus 0.17) and even lower SBF (0.749 PLUS OR MINUS 0.02). Furthermore, there was a negative correlation (r = - 0.652) between the mean arterial pressure and the SBF when results for all patients were considered. The correlation remained (r = - 0.568) in the EH group and the slope of regression line was not different from that for all subjects. The CI and SBF were weakly correlated (r = 0.423) in control subjects and patients with EH, whereas in patients with RAS, a negative correlation was found (r = - 0.778). This study indicates that the SBF, although significantly decreased in patients with EH, remains proportional to the CI in control subjects and in essential hypertensive patients. No redistribution of CI in regard to the splanchnic circulation occurs in EH. In contrast, in patients with RAS a dissociation of CI and SBF occurs and the fraction of the CI which passes through the splanchnic vascular bed is markedly reduced. The close correlation between mean arterial pressure and SBF suggests that both parameters are influenced by a common pathophysiological factor.


De subitaneis mortibus. XII. Asymmetrical hypertrophy of the heart.

Subjects with asymmetrical hypertrophy of the heart are prone to sudden death. Neither the pathogenesis of the eccentric hypertrophy nor the mechanism of sudden death is fully understood. In this report we describe certain postmortem findings in the hearts of 22 subjects who died suddenly, silently and unexpectedly, and in whom the only significant abnormality at autopsy was asymmetrical hypertrophy of the heart. Deep clefts were present in the septum in seven hearts, the small coronary arteries were abnormally narrowed in ten, the sinus node was sclerosed by fibrosis in 12, there was variable narrowing of the atrioventricular (A-V) node artery in many and the His bundle was too thin in three. There were multiple cysts or channels in the central fibrous body and of the adjacent A-V node and His bundle in four hearts. Most of the hearts displayed a fetal dispersion of the A-V node and His bundle throughout the central fibrous body, but this was particularly conspicuous in 13 hearts. These abnormalities in all parts of the conduction system suggest a variety of possible mechanisms by which the heart could become electrically unstable but do not indicate that one single mechanism is at fault in all. They offer some explanation for the reported high incidence of atrial fibrillation in such patients, and why they fare so badly with this arrhythmia. While the pathogenesis of asymmetrical hypertrophy may in some part be attributable to narrowed small coronary arteries or to an abnormal sequence or speed of septal and ventricular activation or to mechanical deficiency caused by deep septal clefts, none of these features was universally present in our series. Both asymmetrical hypertrophy of the heart and the sudden death which so frequently accompanies it probably develop by a variety of pathogenetic mechanisms.


Hypertensive pulmonary vascular disease associated with patent ductus arteriosus. Primary or secondary?

In an infant with patent ductus arteriosus and pulmonary hypertension, elevated pulmonary vascular resistance persisted following ligation of the ductus. Histologic examination of a pulmonary biopsy at two years of age and tissue obtained at autopsy at three years of age showed obstructive pulmonary vascular disease. The question as to whether the organic vascular lesions are secondary to the effects of the patent ductus or part of pulmonary hypertension cannot be resolved. The age of the patient favors a primary etiology.


Comparison of five radioimmunoassays and enzyme bioassay for measurement of digoxin in blood.

Digoxin-containing sera from 86 patients were analyzed for this drug by bioassay and radioimmunoassay. Each serum was analyzed in duplicate by six procedures: inhibition of Na+-K+-dependent ATPase and five "kit" radioimmunoassays from four different commercial sources. Mean values for two of the radioimmunoassays differed significantly from those for the bioassay. One radioimmunoassay mean value was significantly different from the other five mean values. We conclude that normal values for digoxin radioimmunoassay should be determined for each kit, and should not be adopted from published data.


A radioimmunoassay for isoniazid.

A sensitive radioimmunoassay is described, for measuring isoniazid in therapeutic and subtherapeutic concentrations. This radioimmunoassay was used in an exploratory clinical study to measure, as a function of time, the concentration of this drug in plasma of patients receiving isoniazid therapy.


Genetic aspects of orthopedic conditions.

Genetic conditions seen by orthopedists may be divided into Mendelian disorders, chromosome abnormalities and multifactorial conditions. Mendelian disorders involve the abnormality of a single gene and obey the rules of Mendelian inheritance. Chromosome abnormalities are caused by the absence or duplication of a sufficient number of genes to allow this abnormality to be detected by chromosome studies. Chromosome studies are used to confirm the diagnosis of well described syndromes of deletion or trisomy. These studies are also necessary to determine whether mongolism has been caused by non-disjunction or translocation, so that proper genetic counseling can be carried out in these conditions. Multifactorial conditions are determined by several genes or more commonly, by a combination of genetic and environmental factors. These conditions recur in families in a greater incidence than one would expect in the general population but do not obey the rules of Mendelian inheritance. Genetic counseling cannot be performed without an absolute diagnosis, an absolute knowledge of the patterns of inheritance and a well documented family pedigree. The basic principles of genetics covered in this article should allow the orthopedist to understand when genetic counseling is indicated for his patients.


Developmental programming for retinotectal patterns.

Programming events at stages 28-31 in Xenopus specify the cellular positional information that individual retinal ganglion cells will use to derive theri appropriate locus specificity for assembly of the retinotectal map.  The 'programme' that emerges in the stage 31 retina affects the entire ganglion cell population (99 percent of which is generated later) and refers positional information to intraretinal axes (AP and DV) and zero-points.  Its expression in intact retinae was not modified by repeated reintroduction into pre-stage 28 orbits, prolonged eye culture in vitro, or severe disruption of the timing and sequence of ganglion cell births or of optic fibre arrivals in the tectum.  In contrase, intraretinal reorganizations did not produce major modifications (e.g. after transection, partial ablation, fragment fusion etc.) in the set of locus specificities arising in various retinal regions and, in some instances, in the reference axes themselves.  The modified programmes were characteristic of the components undergoing reorganization, but were convergent (many sets of reorganizing components gave a few final patterns); they appeared to involve a stable and rapid reprogramming of certain components by others, in a hierarchical fashion.  The remaining experiments focus on the problem of localizing the 'trigger' for the transition from the unspecified to the specified state in the retina at stages 28-31.  Specification can occur in vitro, based on reversible AP and DV orientational markers which are present in the pre-stage 28 eye primordium; in heterochromically grafted eyes, specification was neither precipitated nor delayed by altering the stage of the host.  Finally, chemical dissection of the differentiating eye primordium confirmed the inference (from Jacobson's [3-H] thymidine labelling kinetics) that a specific gangliogenic precursor cell type exists in the stage 28 retinal neuroepithelium, and suggested that differentiative events in these cells trigger the specification process.


[Fatty-acid pattern of human fat and liver tissues in alcoholics and non-alcoholics: fatty-acid pattern as a measure of alcoholism (author's transl)].

Samples from the right lobe of the liver and from fat tissue of the abdominal wall were removed shortly after death from 116 persons. Fatty-acid pattern of both tissues (lauric, myristic, palmitic, palmitoleic, stearic, oleic, linoleic acids) was measured and compared with the degree of fatty infiltration of the liver. The data were also correlated with chronic excess alcohol consumption in the pre-terminal phase. It was found that with increasing fatty inflitration there was a rise in the relative proportion of palmitoleic acid in liver and subcutaneous fat tissues, with a fall in the relative proportion of stearic acid in the latter. Comparison of results in alcoholics (19 subjects) and non-alcoholics (91) showed the expected higher fatty infiltration of the liver in the former (x equal 13.4% and 5.6%, respectively). Both in liver and subcutaneous fat tissues the relative proportion of palmitoleic acid was significantly higher in alcoholics. But while in these the proportion of palmitoleic acid in the liver was higher than in subcutaneous tissue, the relationship was the converse among the non-alcoholics.


[Myotonic dystrophy (urological features and histochemical findings in muscle) (author's transl)].

Dilatation of the renal pelvis, ureters and bladder appeared in two brothers with myotonic dystrophy (aged 26 and 29 years), probably as a result of involvement of the smooth muscles of these organs. Histochemical differentiation of skeletal muscle fibres (main fibre types I and II, after Engel) indicated a predominant size reduction in type I fibres on demonstrating myofibrillary ATPase. In addition, demonstration of this enzyme also revealed a marked shift of the fibre type proportions in favour of type I. Isolated reduction in gammaG-globulins was also demonstrated. A defect of cellular and humoral response was excluded.


[Evaluation of econazol in 594 cases of skin mycosis (author's transl)].

A total of 594 patients were treated with the new antimycotic agent econazol (Pevaryl). The diagnosis was proven microscopically and on culture except in cases of pityriasis versicolor where it was proven microscopically only. Econazol was given to 130 patients as a 1% solution, to 128 patients as a 1% milk, and to 336 patients as a 1% spray solution. In 333 cases of foot mycosis or eczema marginatum econazol spray powder was given in addition. As measured by the cure rate the spray solution (92%) was not significantly more effective than the milk (89%) or the simple solution (87%). Criteria for cure included negative microscopy and fungal cultures a week after treatment had ceased as well as clinical cure. A total of 90% of all cases (n = 536) could be considered as cured microscopically and on culture after an average of 3.6 weeks (pityriasis versicolor) and 4.8 weeks (tinea pedis, manus, inguinocruralis). In a further 4.4% (n = 26) the fungus could be demonstrated microscopically despite a clinical cure and in 7 of these cases culture was also positive. The cure rate was independent of the responsible pathogen. The preparations were tolerated extremely well. In 7 cases, however, transient dermatitic irritations were seen in the inguinocrural region, mainly caused by the simple solution.


Macrophage metabolism during phagocytosis and digestion of normal and IgG antibody-coated sheep erythrocytes.

The respiration rate, aerobic glycolysis and glycolytic enzyme activities of purified mouse peritoneal macrophages increase during phagocytosis of either free or antibody-coated SRBC. These changes, however, seem to be roughly parallel to the amount of phagocytized antigen. In the digestive phase, Ag-Ab complexes induce more marked changes in macrophage metabolism than a comparable amount of free antigen. Lactate production, activity of several glycolytic enzymes and alanine-aminotransferase as well as glycogen content are greatly affected. Since in cell transfer experiments macrophage-associated SRBC can prime normal recipients for antibody response, whereas cell-associated SRBC-antibody complexes cannot, it is concluded that the changed macrophage metabolism drives antibody-coated antigen into metabolic channels allowing its rapid and total degradation.


Studies on intestinal adenosine triphosphatases. I. Application of a semiautomated method to the rat intestinal brush borders.

Elaboration of a semiautomated kinetic test on LKB 8600 apparatus for ATPase is described, using the PK-LDH system. As optimal ionic conditions 3 mmol-1 - minus 1 potassium chloride and 100 mmol-1 - minus 1 sodium chloride are proposed for measurement of (Na+-K+)-ATPase activities of rat intestinal brush borders. NH+4 can substitute for K+. The coefficients of variation of the method are 2.4% for Mg2+-ATPase and 4.9% for (Na+-K+)-ATPase determinations.


Congenital aneurysm of the left atrium. Case report and analysis of the literature.

A congenital aneurysm of the left atrium is an unusual disease, which can be confused with other diseases of the heart and of the mediastinum. It is a frequent cause of complications, like dysrhythmias and embolism. A surgically treated case is reported.


[Isolation of a new keto-acid, produced from epsilon-N-trimethyllysine by Neurospora crassa].

A new epsilon-N-trimethyllysine metabolite has been isolated from the mycelium of Neurospora crassa. The labelled compound produced from incubations in vivo and in vitro from epsilon-N-trimethyl ([14-C]H3)L-lysine has been identified as 2-keto-epsilon-N-trimethyl-hexanoic acid by reducing its 2,4-dinitrophenyl hydrazone back to epsilon-N-trimethyllysine by hydrogenolysis in a Parr bomb. Analyses on TLC and in four different ion exchange chromatographic systems show the appearance of a ninhydrin positive product having the same Rf and the same retention time as epsilon-N-trimethyllysine; it contains more than 85% of the radioactivity of the 2,4-dinitrophenylhydrazone of the keto acid.


Antibiotic inhibitors of mitochondrial ATP synthesis.

Fourteen antibiotics have been found to inhibit oxidative phosphorylation and uncoupler-stimulated adenosinetriphosphatase in mitochondria. Four different types of binding sites for these inhibitors have been found. The first (1) binds aurovertin to purified MF1 ATPase in the stoichiometric ratio of two aurovertin molecules per molecule of ATPase. Site II is the locus for efrapeptin (A23871) and may be a catalytic site on purified ATPase. The remaining two sites have been demonstrated only in mitochondria or submitochondrial particles when the APTase is bound to other membrane components. Oligomycin, venturiciden, venturicidin X and ossamycin probably all bind at site III. Leucinostatin (A20668) binds at site IV. At low concentrations, this antibiotic acts like oligomycin; at higher concentrations it uncouples oxidative phosphorylation. Venturicidin appears to prevent leucinostation from binding at site IV for it allows uncoupling to occur at very low concentrations of the latter antibiotic. Venturicidin aglycone, which is a more effective inhibitor than its parent compound, does not exert this effect. It is concluded that sites III and IV are in juxtaposition and that when venturicidin binds at site III its sugar moiety projects into the area of site IV to prevent leucinostation from binding at its inhibitory site.


Coupling of "high-energy" phosphate bonds to energy transductions.

Recent results suggest consideration of a new concept for oxidative phosphorylation in which a prime function of energy is to bring about release of ATP formed at the catalytic site by reversal of hydrolysis. Data with submitochondrial particles include properties of an uncoupler insensitive Pi=HOH exchange, a rapid reversible formation of bound ATP in presence of uncouplers, and predictable patterns of 32-Pi incorporation into ATP in rapid mixing experiments. ADP is confirmed as the primary Pi acceptor in mitochondrial ATP synthesis, but with chloroplasts ADP is also rapidly labeled. Other findings with pyrophosphatase and  with transport ATPase harmonize with the new concept. Measurements of the reversal of ATP cleavage and binding by myosin suggest that oxygen exchanges result from reversible cleavage of ATP to ADP and Pi at the catalytic site and that the principal free energy change in ATP cleavage occurs in ATP binding. Reversal of conformational changes accompanying ATP binding and cleavage is proposed to drive the actin filament in contraction. Thus energy transductions linked to ATP in both mitochondria and muscle may occur primarily through protein conformational change.


Chloroplast membranes and coupling factor conformations.

The demonstrated role of proton translocation and resulting electrochemical activity gradients (protonmotive force) in ATP synthesis by chloroplasts is noted. Evidence for the participation of conformational changes in the terminal ATPase (coupling factor, or CF1) is reviewed. Hydrogen exchange into ordinarily cyptic groups of the molecule occurs only when the subtending membranes are put under the stress of a protonmotive force. Since up to 100 hydrogen atoms per mole are involved in the energy-dependent exchange the conformational change permitting tham access to the medium must be a major one. Chemical reagents are beginning to be used to attack groups on CF1 that are exposed only when the membranes are energized. N-ethylmaleimide binds covalently, sulfate causes as yet unspecified damage, and permanganate leads to oxidative damage to CF1 under energized conditions. The last two reagents are analogues of phosphate, and ADP must be added for them to inhibit. On the basis of this and other differences between the conditions needed for inhibition by permanganate or sulfate, and that by N-ethylmaleimide or the hydrogen exchange, a somewhat complex scheme involving several successive or alternative conformations of CF1 can be postulated. Questions are raised as to the way in which a conformational change in a bound protein could be caused by a proton activity gradient across its supporting membrane, and as to whether the altered conformations might constitute a part of the energy transformations leading to ATP synthesis.


Laparoscopic recovery of mature human oocytes.

This report provides a detailed description of a method by which mature or maturing human oocytes can safely be recovered by follicular aspiration during laparoscopy. By combining gonadotropin administration with laparoscopic recovery, oocytes which have resumed or completed meiotic division can be obtained for study without subjecting the patient to more hazardous surgical procedures. Fifteen oocytes were obtained from sixteen patients. On electron microscopic examination, four were found to have extruded polar bodies and were therefore classified as mature. Daily rather than intermittent administration of HMG, larger HMG dose, longer HCG aspiration intervals, and controlled aspiration pressures appeared to be the factors which produced the best yield of mature oocytes.


[Comparison of temperature measurements during laparoscopic sterilization and the histologic findings of the cauterized segments of the fallopian tube (author's transl)].

During laparotomies the temperature generated by tubal sterilization with the laparoscopic equipment was measured continually in 32 fallopian tubes. The thermal damage to the fallopian tube caused by different energies and coagulation durations was examined microscopically. During the coagulation of the fallopian tube, with an energy of 20-30 watt temperatures of around 100 degrees C are produced at the point of coagulation. The effective coagulation time is dependant upon the energy created and the duration of the coagulation. The best histologic results were obtained with a duration of the coagulation of 60 seconds at an energy of 30 watts. This corresponds to an effective coagulation time of 50 seconds. The temperature of the coagulation instrument becomes normal within 1 to 2 minutes after the coagulation. If the electric energy is increased to 50 or 100 watt, sparking is observed almost consistently. With these energies, temperatures up to 600 degrees C are observed. The temperature of the metal parts of the coagulation forceps then become normal after 3-4 minutes. In order to avoid burns during laparoscopic tubal sterilizations and in order to obtain a good histologic result the procedure should be done with an electrical energy of 30 watt and a duration of the coagulation of 60 seconds.


Heterokaryon incompatibility genes in Neurospora crassa detected using duplication-producing chromosome rearrangements.

Evidence is presented for five or six previously undetected heterokaryon incompatibility (het) loci, bringing to about ten the number of such genes known in Neurospora crassa. The genes were detected using chromosome duplications (partial diploids), on the basis of properties previously known for het genes in duplications. Duplications homozygous for het genes are usually normal in growth and morphology, whereas those heterozygous are strikingly different. The heterozygotes are inhibited in their initial growth, produce brown pigment on appropriate medium, and later "escape" from their inhibition, as a result of somatic events, to produce wild-type growth. - Five normal-sequence strains were crossed to 14 duplication-producing chromosome rearrangements, and the duplication progeny were examined for properties characteristic of duplications heterozygous for known het genes. Each cross produced duplications for a specific region of the genome, depending on the rearrangement. Normal-sequence strains were wild types from nature, chosen from diverse geographic locations to serve as sources of genetic variation. - The duplication method was very effective. Most of the longer duplications uncovered het genes. The genes are: het-5 (on linkage group IR, in the region covered by duplications produced using rearrangement T (IR LEADS TO VIR)NM103), het-6 (on IIL, covered by T(IIL LEADS TO VI)P2869 and T(IIL LEADS TO IIIR)AR18 duplications), het-7 (tentatively assigned to IIIR, T(IIIR LEADS TO VIL)D305), het-8 (VIL, T(VIL LEADS TO IR)T39M777), het-9 (VIR LEADS TO IVR)AR209), and het-10 (VIIR, T(VIIR LEADS TO IL)5936.


The use of duplication-generating rearrangements for studying heterokaryon incompatibility genes in Neurospora.

Heterokaryon (vegetative) incompatibility, governing the fusion of somatic hyphal filaments to form stable heterokaryons, is of interest because of its widespread occurrence in fungi and its bearing on cellular recognition. Conventional investigations of the genetic basis of heterokaryon incompatibility in N. crassa are difficult because in commonly used stocks differences are present at several het loci, all with similar incompatibility phenotypes. This difficulty is overcome by using duplications (partial diploids) that are unlikely to contain more than one het locus. A phenotypically expressed incompatibility reaction occurs when unlike het alleles are present within the same somatic nucleus, and this parallels the heterokaryon incompatibility reaction that occurs when unlike alleles in different haploid nuclei are introduced into the same somatic hypha by mycelial fusion. - Nontandem duplications were used to confirm that the incompatibility reactions in heterokaryons and in duplications are alternate expressions of the same genes. This was demonstrated for three loci which had previously been established by conventional heterokaryon test-het-e, het-c and mt. These were each obtained in duplications as recombinant chromosome rearrangements. The particular method of producing the duplications is irrelevant so long as the incompatibility alleles are heterozygous. - The duplication technique has made it possible to determine easily the het-e and het-c genotypes of numerous laboratory and wild strains of unknown constitution. In laboratory strains both loci are represented simply by two alleles. Analysis of het-c is more complicated in some wide strains, where differences have been demonstrated at one or more additional het loci within the duplication used and multiple allelism is also possible. - The results how that the duplication method can be used to identify and map additional vegetative incompatibility loci, without the necessity of heterokaryon tests.


The response of the right ventricle to experimentally induced pulmonary artery obstruction.

Banding of the main pulmonary artery was undertaken in 11 litter-mate puppies and 8 young adult beagle dogs to study the response of the right ventricle to pressure loading. Five puppies and 3 young adult dogs acted as controls, undergoing sham surgical procedures without pulmonary artery banding. In one banded puppy and one banded young adult dog, adequate banding was not achieved and each was subsequently considered with its respective control group. A trans-band gradient of 15 mm Hg or greater was deemed a successful banding procedure. Banded puppies showed an increase in the trans-band pressure gradient in the year following surgery whereas young adult dogs showed a fall in the gradient, despite satisfactory banding as shown by cardiac catheterization 14 days after surgery. Cardiac muscle analyses indicated that the right ventricle of puppies responded to pulmonary banding by an increase in total organ weight, total tissue protein, DNA and RNA without altering the protein/DNA and RNA/DNA ratios. By contrast, young adult dogs showed only slight and insignificant increases in the protein/DNA and RNA/DNA ratios. Thus the response of the right ventricle to pulmonary artery banding in puppies is hyperplastic in nature, while in young adult animals the response to a similar stress load is with mild muscular hypertrophy and dilatation. This differential cellular response to the same stimulus based on the age of the organism is another demonstration of the principle that growth responses in the young involve cellular proliferation while those in the adult consist primarily of cell enlargement. It is, therefore, perhaps erroneous to refer  to cardiac enlargement in newborns and infants as cardiac hypertrophy.


[Hepatic allopurinol oxidizing enzyme in mice].

The relationship between allopurinol oxidizing enzyme and aldehyde oxidase was investaged in mice. The oxidation of both N-methylnicotinamide and allopurinol appears to be catalized by a single enzyme, aldehyde oxidase (aldehyde-oxygen oxidoreductase EC, 1.2.3.1.). This conclusion is based on the following evidence; The postnatal changes of allopurinol and N-methylnicotinamide oxidizing activities were similar during growth and the levels of both activities increased in a parallel fashion upon the attainment of sexual maturity. The rates of loss of the activities of both enzymes by heat denaturation as well as dexamethasone administration were similar. The inhibitors of allopurinol oxidizing enzyme also suppressed N-methylnicotinamide oxidation. Competition of N-methylnicotineamide and allopurinol for oxidation was demonstrated. The rate of increase of the activities in both enzymes was almost parallel during each step of the purification from mouse liver supernatant. It was ascertained that xanthine oxidase in the enzyme preparation does not influence allopurinol oxidation.


Pathologic anatomy of the cardiomyopathies. Idiopathic dilated and hypertrophic types, infiltrative types, and endomyocardial disease with and without eosinophilia.

This presentation summarizes necropsy observations in patients with three types of cardiomyopathy: idiopathic, infiltrative, and endomyocardial disease. The idiopathic variety is subdivided into two types depending on the size of the ventricular cavity. In the dilated ventricular type the left ventricular wall is frequently less than 1.5 cm. thick, intracardiac thrombi are common, the atrioventricular valve rings usually are mildly dilated, and focal myocardial and endocardial scars are common. In the nondilated type (hypertrophic cardiomyopathy), the ventricular septum is usually thicker than the left ventricular free wall, which also is thick (greater than 1.5 cm.). When the septum is similar in thickness to the left ventricular free wall (symmetric), left ventricular outflow obstruction does not occur. When the septum is thicker than the left ventricular free wall (asymmetric), left or right ventricular outflow obstruction may or may not be present. The orientation of myocardial fibers one to another in the ventricular septum in the nondilated (hypertrophic) type is abnormal, whereas it is normal in the dilated ventricular type. Intracardiac thrombi are rare and atrioventricular valve rings are never dilated in the nondilated type of idiopathic cardiomegaly. The infiltrative types of cardiomyopathies include iron, calcium, lipids, mucopolysaccharides, granulomas, amyloid, and neoplasms. The first four usually are located within myocardial cells and the latter three, between myocardial cells. It is probable that all these myocardial infiltrates are capable of producing cardiac dysfunction, primarily on a restrictive basis. Endomyocardial disease may or may not be associated with eosinophilia. When the latter occurs, the eosinophils are structurally normal. Death is related to congestive cardiac failure. This category is actuality also in idiopathic.


Immunohistochemical findings in the renal vascular lesions of progressive systemic sclerosis.

Four selected patients with progressive systemic sclerosis showed deposition of immunoglobulins and complement in diseased renal arteries and arterioles. Although three patients had hypertension, in two of these malignant, the third patient did not have hypertension over a four year period. These findings suggest that immune complexes may be involved in the pathogenesis of some cases of progressive systemic sclerosis, the primary target being the vascular system.


Reproduction in a female patient with Down's syndrome. Case report of a 46, XY child showing slight phenotypical anomalies, born to a 47, XX, + 21 mother.

A non-mongoloid boy born to a mongoloid mother is described. He showed aplasia of the left 5th finger and some clinical and dermatoglyphic features frequently found in Down's syndrome. Chromosome analysis revealed few hyperdiploid but no G-trisomic cells. An undetected G-trisomy mosaic, or a mechanism of extrachromosomal inheritance, and an embryonic development in a pathological milieu are discussed.


Antigenicity of trophoblast and possible antigen-masking effects during pregnancy.

The interaction between cultured human trophoblast cells and materanl lymphocytes was used as an in vitro model to investigate trophoblast antigenicity. Cytotoxic effects in the trophoblast monolayer were apparent after 72 hr incubation and depended on the presence of non-lymphoid cell types in addition to lymphocytes. Lysis of trophoblast was preceded by blast cell formation and apparently involved close contact between the maternal cells and the trophoblast cells. The nature of the cytotoxic reaction suggested the presence of histocompatibility factors on trypsinized trophoblast cells. This manifestation could be due to removal of fibrinoid or enhancing antibody from the cultured cells. The ability of trypsinized trophoblast cells to synthesize mucoprotein was investigated by the Hale colloidal iron test and found to be unimpaired. The effect of maternal serum on the recognition of human trophoblast antigens by maternal lymphocytes was used as an in vitro model to investigate the occurrence of enhancing antibody in maternal serum. The cytotoxic effects of maternal lymphocytes on trophoblast were completely prevented by the presence of maternal serum, this protective effect being reduced significantly by removal of IgG from the maternal serum. A slight protective effect of allogeneic pregnancy serum was also observed. It is suggested that these findings support a role for immunological enhancement in maintaining the foetal allograft.


Subpopulations of mouse spleen lymphocytes. III. Cellular interactions in the response to concanavalin A.

The profile of response to concanavalin A (con A) of purified mouse T cells was found to differ appreciably from that of non-fractionated spleen cells, in agreement with results previously published by other investigators. Experiments designed to elucidate the reasons underlying these differences have revealed that the response of the spleen cells to con A is determined by a complex interplay between several cell types. (a) B cells contribute to the overall incorporation of thymidine in the presence of con A-stimulated T cells. However, the B cells participate in the response only if the T cells are dividing. (b) A population of 'adherent cells' is present in the spleen, which enhances the stimulation of the spleen cells by low doses of con A but suppresses the response to high doses of mitogen. These adherent cells include most likely the conventional macrophages, but probably also a population of 'suppressor T cells'. (c) Such 'suppressor T' cells can be readily detected among the peritoneal exudate cells. Addition of the exudate cells to cultures of purified T cells enhances the response to low doses of con A. This effect can be further increased by treating the peritoneal cells with a cell T-specific antiserum and complement, i.e. by eliminating the T cells.


Dynamic viscoelastic study of the effect of beta-actinin on the interaction between F-actin and heavy meromyosin.

Beta-Actinin, a regulatory protein of muscle, greatly decreased the dynamic rigidity modulus of an acto-heavy meromyosin solution. However, it was found that the mechanical mixing procedure used resulted in fragmentation of the decorated F-actin particles and beta-actinin inhibited the reassociation of the fragmented particles. This was the reason for the decrease of the dynamic rigidity modulus of the acto-heavy meromyosin complex caused by beta-actinin. When beta-actinin was added to the acto-heavy meromyosin solution in the presence of ATP, it did not affect the visco-elasticity. F-actin particles dissociated by ATP were not fragmented by the mixing procedure, which was responsible for the apparent inactivation of beta-actinin in the presence of ATP.


[Preparative electrophoresis of peptidic glycosaminoglycans. Application to the fractionation of peptidic glycosaminoglycans of arterial wall (author's transl)].

A new procedure for the fractionation of glycosaminoglycans by electrophoresis on Pevikon has been described. Mixtures of glycosaminoglycans were fractionated by preparative electrophoresis on Pevikon in pyridine formate or glycine-HCl buffers. By this procedure, 200 mg of a mixture of hyaluronic acid, heparan sulphate, dermatan sulphate and chondroitin sulphate isolated from arterial wall could be successfully separated without loss of material. The purified fractions were analysed by enzymatic and chemical procedures. The molar ratios of uronic acid to hexosamine and of sulphate to hexosamine and the amino acid content of each glycosaminoglycan have been determined. The peptidic content (less than 1%) is represented by five amino acids, viz. serine, glycine, alanine, aspartic acid, and glutamic acid.


Hepatic mitochondrial function in ketogenic states. Diabetes, starvation, and after growth hormone administration.

The study was designed to evaluate hepatic mitochondrial function during ketotic states. The ketogenic models studied were streptozotocin-induced diabetic ketoacidosis, 48 h of starvation, and after growth hormone administration. In the last-mentioned model we observed increased free fatty acids but not ketonemia. Oxidative phosphorylation was measured using the citric acid cycle substrates pyruvate and succinate, the amino acid glutamate, a ketone body beta-hydroxybutyrate, and a long-chain fatty acid palmitoyl-l-carnitine. State 3 (ADP stimulated) and state 4 (ADP limited) respiration, respiratory control ratio (state 3/state 4), and the ADP/O ratios were normal in the controls and the experimental groups. Uncoupled respiration produced by dinitrophenol with a variety of substrates was unchanged in the experimental groups compared to the controls. Fatty acid oxidation was studied in detail. The rate of utilization of palmitoyl-l-carnitine by controls or experimental groups did not depend on the product formed (citrate, acetoacetate). No significant changes were observed in the oxidation of palmitoyl-CoA (+ carnitine) or with an intermediate-chain fatty acid hexanoate. The specific activity of hepatic mitochondria carnitine palmitoyltransferase did not change in any of the three experimental groups. It is concluded that during diabetic ketoacidosis, starvation, and growth hormone administration, there is (a) no alteration in hepatic mitochondrial function; (b) no change in the intrinsic capacity of hepatic mitochondria to oxidize fatty acids; and (c) no change in the specific activity of mitochondrial carnitine palmitoyltransferase. The mechanism by which the body restrains flux through the mitochondrial oxidative machinery remains to be fully determined.


Effects of insulin and dietary myoinositol on impaired peripheral motor nerve conduction velocity in acute streptozotocin diabetes.

The factors influencing the development of impaired sciatic motor nerve conduction velocity (MNCV) in acute experimental diabetes were examined. Decreased MNCV developed by the 14th day after streptozotocin administration but only in rats which became hyperglycemic. Insulin treatment, begun on day 3, failed to prevent imparied MNCV in diabetic rats in which improved or normal weight gain and a decreased degree of hyperglycemia was induced. However, insulin treatment prevented the development of impaired MNCV in a group of diabetic rats in which the tail vein plasma glucose concentration was never found to exceed 160 mg/dl during days 6 through 14, andin which the mean plus or minus SEM of the average plasma glucose concentration for each animal during the same period was 75 plus or minus 18 mg/dl. In normal rats fed diets containing 0.011% or 0.069% free myoinositol (a presumably normal range), sciatic nerve free myoinositol concentrations were 90- and 60-fold higher than those in plasma. On these diets the development of impaired MNCV in the diabetics was associated with a decrease in nerve free myoinositol as compared with nerves from normals fed the same diet, despite similar plasma levels in the normals and diabetics. Plasma and nerve free myoinositol increased with increasing dietary myoinositol content in both normals and diabetics, and nerve myoinositol content could be acutely increased by an i.p. myoinositol load. By supplementing the diets with 1.0% myoinositol, the difference in nerve myoinositol in normal and diabetic rats on day 14 was abolished; on this diet the development of impaired MNCV in the diabetics was moderated or totally prevented, despite persistent hyperglycemia and elevated nerve sorbitol and fructose concentrations. Insulin treatment that prevented impaired MNCV prevented a decrease in nerve myoinositol in diabetics. These studies suggest that insulin deficiency, and possibly hyperglycemia, are primary factors in the development of imparied MNCV in acute experimental diabetes. However, the development of impaired MNCV appears to be related in some manner to a derangement in the regulation of nerve free myoinositol content, which appears to be subject to modification by increases in plasma myoinositol concentration over a critical range.


Is the macrophage the stimulating cell?

Using CAF1 spleen cells to stimulate parental strain BALB/c spleen cells in a mixed lymphocyte culture, column separation of responding cells increased their response whereas the same treatment of stimulating cells reduced their activity approximately 95 per cent. Peritoneal macrophages from CAF1 mice were found to stimulate BALB/c spleen cells poorly if present in comparable numbers or if they were cultured for 24 hours before adding responding cells. However, if the F1 macrophages were in contact with the BALB/c cells for only 4 hours, their stimulating effect was increased strikingly. Under these conditions BALB/c macrophages had no effect. It is concluded that the macrophage is probably the most effective stimulating cell and may be the only cell with this capability.


Streptozotocin-induced diabetes mellitus and the host-parasite relation in murine schistosomiasis mansoni.

The effect of streptozotocin-induced diabetes mellitus on both the host and the parasite was studied in mice infected with Schistosoma mansoni. Neither the drug nor the marked rise in concentration of blood glucose had any effect on penetration of the skin by the cercariae, their subsequent maturation into adult worms, or their output of eggs. During the acute stages of the infection (at eight weeks), the diabetic animals showed marked suppression of the host granulomatous reaction to schistosome eggs trapped in the liver, accompanied by an alleviation of hepatosplenic disease. During the chronic stages of infection (at 16 weeks), there was a pronounced megalocytosis of the hepatocytes only in the infected animals with streptozotocin-induced diabetes; these animals also had an exacerbation of hepatosplenic disease.


Chronic mucocutaneous candidiasis with macrophage dysfunction, a plasma inhibitor, and co-existent aplastic anemia.

A 56-year-old man developed chronic mucocutaneous candidiasis (MCC) and pernicious anemia. Nine years later he developed aplastic anemia which ultimately was fatal. A small thymoma was found at autopsy. He was anergic and his mononuclear leukocytes (MNL) failed to undergo a proliferative response in culture to soluble antigens. His monocytes did not mediate a proliferative response by lymphocytes from sensitized control donors when stimulated with Monilia albicans antigen but did mediate a mixed leukocyte reaction normally. His plasma contained a poten inhibitor of -3H-thymidine incorporation by sensitized control MNL when stimulated with soluble antigens but was not inhibitory of the mixed leukocyte reaction, lymphoproliferative responses to plant mitogens, and was not shown either in vivo or in vitro to depress hematopoiesis. Patient lymphocytes were responsive to plant mitogens, Monilia antigen, in the mixed leukocyte reaction, and produced macrophage migration inhibitory factor in response to Monilia antigen. After plasmapheresis, delayed hypersensitivity and lymphoproliferative responses to soluble antigens were temporarily restored. This case implicates the macrophage in the pathogenesis of MCC and demonstrates some consequences of chronic monocyte dysfunction. The inhibitor of some expressions of cell-mediated immunity was removed by plasmapheresis.


Metabolic clearance rate, production rate and mammary uptake of progesterone in the goat.

The dynamics of progesterone uptake and metabolism in the mammary gland of the goat have been measured and related to the metabolic clearance rate and production rate of the hormone determined by tracer kinetic techniques. The metabolic clearance rate of progesterone from blood was 3-13 plus or minus 0-35 (S.E.M.) 1/min in ten experiments on six goats; values tended to be slightly higher in pregnant than in non-pregnant goats. The production rate of progesterone at oestrus, and at day 3 of the normal cycle, was less than 0-01 mug/min. During the luteal phase of the oestrous cycle the production rate was 8-5 and 14-6 mug/min in 2 animals, and in the second half of pregnancy, 15-3 plus or minus 0-6 mug/min (5 animals). Progesterone was extracted from the circulation by the mammary gland of conscious goats with an efficiency of 49-4 plus or minus 11-3% in non-pregnant, and 51-7 plus or minus 11-5% in pregnant aniamals. The mean clearance rate of progesterone by the udder was 0-279 1/min, 8-8% of the metabolic clearance rate. Mammary uptake of progesterone in goats with an actively secreting corpus luteum was 0-64 plus or minus 0-29 mug/min, which gave an estimated value of 0-11-1-88 ng/min/g mammary gland. The mammary extraction of progesterone was investigated in a goat 3 days after oestrus when any high affinity receptor sites would presumably be unoccupied. During the infusion of progesterone into a mammary artery, tissue sample were taken from various organs, including the mammary gland, and the concentration of labelled compounds at steady state was determined. A high mammary extraction of progesterone was found to be determined. A high mammary extraction of progesterone was found to be attributable principally to progesterone metabolism. The metabolites of progesterone were removed from the gland in venous blood and were not stored to any appreciable extent in mammary tissue. Experiments in vitro confirmed the findings in vivo that mammary tissue metabolized labelled progesterone and also pregnenolone and androstenedione; metabolism of dehydroepiandrosterone, oestradiol-17 beta, oestrone and cortisol was relatively small. Confirmation of our previous finding that the mammary gland of the goat can synthesize progesterone from labelled pregnenolone infused into the gland in vivo, further implicated this organ as an active site of metabolism of certain steroids. The physiological role of steroid metabolism in the mammary gland is discussed.


Baseline (spontaneous) symptoms in healthy persons--a prospective study.

Many healthy people have unexplainable complaints. Healthy subjects who had not suffered from any disease in the previous two months and who were not taking any drugs during the previous two weeks were interrogated daily for seven days to determine whether they experienced any abnormal feeling (baseline or spontaneous symptoms). Of 398 subjects interrogated, 49 (12.5 per cent) gave a history of having suffered from one or more symptoms. Incidence of baseline symptoms was higher in the medical group compared with the nonmedical group and in females compared with males. In the study of adverse drug reaction, controls for spontaneous symptoms in the population under surveillance are essential.


Inhibitory effects of bilirubin on cellular immune responses in man.

A significant depression in cell-mediated immunity as measured by lymphoproliferative responses to phytohemagglutinin and responsiveness to mixed lymphocyte culture was observed when adult lymphocytes or cord blood lymphocytes were incubated with increasing concentrations of bilirubin. The inhibitory effect of bilirubin could only be demonstrated with suboptimal concentrations of PHA (0.01  and 0.005%) and was more marked in premature infants than in term neonates or adults. This effect was partially reversible after short preincubation with bilirubin, but was more protracted with preincubations of 24 hours or more. Inhibition of MLC responsiveness of 80.1 plus or minus 5.1% was also demonstrated at a bilirubin concentration of 20 mg/dl. Specific cytotoxicity to rubella virus-infected cells, measured by a 51Cr-release microassay, was not found to be depressed. Bilirubin thus appears to have an inhibitory effect on immune responsiveness which is greater on the afferent limb than on the effrent limb of immunity.


The role of gingival connective tissue in determining epithelial differentiation.

Free grafts of connective tissue, without epithelium, were transplanted from  either the keratinized gingiva or the non-keratinized alveolar mucosa (controls) into areas of the alveolar mucosa in seven monkeys. The grafts were placed in pouches created in the connective tissue as close as possible to the overlying epithelium. After 3-4 weeks, the transplants were exposed by removal of the overlying tissue in order to allow epithelialization from the surrounding non-keratinized alveolar mucosa. The transplants were examined clinically and histologically at time peiods between 1 and 12 months. The gingival connective tissue grafts became covered with keratinized epithelium displaying the same characteristics as those of normal gingival epithelium. The alveolar mucosa transplants were covered with non-keratinized epithelium. This indicated that gingival connective tissue is capable of inducing the formation of a keratinized gingival epithelium.


Nuclear size and cytochemical changes during autolysis in free gingival autograft epithelium.

Karyometric and cytophotometric measurements of DNA and RNA have been evaluated in gingival graft epithelium. The study was conducted during the first 20 hours from the time of grafting when the graft epithelium undergoes a process of autolysis. Both basal cell and prickle cell layers were used. The results showed an increase in optical density for Feulgen-DNA according to the time of grafting. At the same time pentose nucleic acid (RNA) and nuclear size showed decreasing values. It was concluded that pycnosis was the usual course of nuclear autolysis in graft epithelium. In addition gingival graft epithelium was found to be an useful model for the study of cell autolysis in vivo.


Complement factors in gingival crevice material from healthy and inflamed gingiva in humans.

The presence and levels of complement factors C3, C4, C5, and C3 proactivator, were determined by electroimmuno assay in gingival cervice material from five individuals with healthy gingiva and from six patients with chronically inflamed gingiva. Higher concentrations of C3 and C4 were found in samples from chronically inflamed gingiva when compared to those healthy gingiva. The amount of C3 in material from healthy as well as from inflamed gingiva was related to that in plasma, when the albumin in plasma and gingival crevice material was used as a reference. The same was found for C4 in samples from healthy gingiva but in those from inflamed gingiva, the values were significantly lower when related to plasma levels. C5 could not be found in material from healthy gingiva but was present in material from inflamed gingiva. C3 proactivator was present in material from inflamed gingiva in the converted form. No C3 proactivator was found in material from healthy gingiva with the methods used. Analysis of C3 in samples from inflamed gingiva, using crossed immunoelectrophoresis, showed that C3 was converted in these samples. The results indicate that the complement system may be activated in gingival crevice material from inflamed gingiva.


Early formation of dental plaque on platic films. 1. Light microscopic observations.

In human subjects with healthy gingiva thin transparent plastic films were applied to the buccal surfaces of premolars in the upper and lower jaws. The films were left in place for peiods of 15, 30, 60, 120 and 240 minutes. The presence of coccoid bacteria, leukocytes and epithelial cells was investigated in an area adjacent to the gingival margin. The results showed that by 15 minutes coccoid bacteria had become attached to the artificial tooth surface. The number of microorganisms gradually increased during the time of the study. Large numbers of cocci and the formation of micro-colonies were observed after 120 and 240 minutes. Increasing numbers of leukocytes and epithelial cells were also found during the period of observation.


Partial-mount recording of plaque and periodontal pockets.

Plaque ananand pocket depths were recorded in milimetres from the tissue margin, coronally or apically, respectively. Plaque and pocket depths were recorded by examining only a segment of the dentition using eight different combination of teeth. These results  d the results obtained from full-mouth examinations in the same individuals were compared. All eight methods of partial-mouth recording gave results that were good in terms of both relative and absolute value when compared to the total-mouth index. The results were valid irrespective of the method chosen. The anterior tooth index was preferred because this segment of the dentition was the easiest to examine. It eliminated the difficulty of making the decision of which teen to substitute when any of the other teeth were missing and produced results for plaque scores and pocket scores that were very good estimates of the total-mouth index.


Periodontal changes in distant teeth due to trauma from occlusion.

The present investigation studies the changes that occur in periodontal supporting tissues of lower canine and premolar of marmosets (Callithrix jacchus), due to experimental modification of occlusal relationships in the region of the molar. For our purposes an extracoronal arch wire of 0.7 mm diameter was fixed into occlusal perforations of the upper third premolar and first molar, on both sides. Eight male marmosets weighing from 250 to 350 grams were used. After each experimental time interval of 7, 14, 21 and 28 days two animals were sacrificed. The results clearly showed areas of tension with stretch periodontal fibers in the distal region of the canine and areas of pressue with osseous and redicular resorption in the mesial region of the first premolar. The tissue changes may be considered to result from transmission of abnormal forces through the proximal contacts of teeth associated with displacements and minor modifications in their occlusal relationships. Similar changes may occur in a tooth distant from the teeth exposed to primary excessive occlusal forces.


The collagen content of skin and gingival tissues in ascorbic acid-deficient monkeys.

Five green monkeys were used in this study.  Three animals were fed a scorbutigenic diet and the remaining two served as controls.  Biopsies were dissected from the vestivular gingiva and dorsal skin and polyvinyl sponges were implanted subcutaneously during 4 week periods throughout the experimental period.  The animals were sacrificed after 12 weeks and the periodontal ligament was removed.  Collagen extracts were prepared from the gingival and granulation tissues and treated with bacterial collagenase.  Following acid hydrolysis the total amount of proline and hydroxyproline was determined in various tissue preparations.  In the tissues examined, the content of hydroxyproline decreased in the scorbutic animals throughout the experimental period.  The decrease in the hydroxyproline content of the gingiva started within the first 4 weeks and was faster than that of the skin, indicating that the extent of decrease is dependent of the turnover rate of the collagen in the tissues.  The synthesis of hydroxyproline was almost totally impaired in the granulation tissue formed in the sponges implanted after the 8th week of experimentation.  Collagenase treatment of collagen extracts resulted in a release of proline and hydroxyproline in a higher ratio in extracts from the experimental animals than in extracts from the controls.  It is concluded that ascorbic acid is a prerequisite for the maintenance of the collagen pool in the tissues and that lack of this vitamin results in the formation of a collagenase degradable protein fraction with a low hydroxyproline content.


Histopathologic features of the initial and early stages of experimental gingivitis in man.

The pathologic alterations occurring in the gingival tissues of humans immediately following the beginning of plaque accumulation have not heen elucidated previously.  Seven males, 22--31 years of age, free of clinical manifestations of dental and periodontal disease, and exhibiting a Plaque Index score of zero for the previous 28 days served as subjects.  Plaque control measures were discontinued for 8 days and biopsies were taken from the buccal marginal gingiva of the first premolars on days 0, 2, 4, and 8.  Paraffin- and Epon-embedded section, treated with a variety of histochemical stains were analyzed microscopically and cell counts were done on 1-micron Epon section.  At 2 and 4 days following the beginning of plaque accumulation, the vessels subjacent to the juctional epithelium exhibited vasculitis and alterations in the perivascular collagen.  There was a statistically significant increase in the number of neutrophils residing in the junctional epithelium.  By the end of the 8-day period, the number of small mononuclear cells, mostly lymphocytes, in the connective tissues had increased by 3-fold and the area of collagen fiber alteration by 4-fold.  In addition, the number of fibroblasts per unit area of connective tissue decreased significantly.  Thus, within the period of 8 days following the beginning of plaque accumulation, an early lesion exhibiting many features characteristic of delayed hypersensitivity develops.


Elevation of a serum component in periodontal disease capable of modulating chemotactic infiltration.

Serum from dental patients was examined for its capacity to affect migration of polymorphonuclear leukocytes toward a complement-independent chemotactic factor, bacterial filtrate of Escherichia coli, (BF).  Seven subjects with advanced periodontitis, documented with clinical measurements of pocket depth, loss of attachment, indices for gingival inflammation and oral hygiene, were paired with normal subjects of similar age and sex.  This study shows that patients whose only apparent clinical symptom was severe periodontal inflammation harbor a heat-stable, serum component which neutralizes factors chemotactic for polymorphonuclear leukocytes.  A role for this component in modulation of inflammatory infiltration is discussed.


In vitro cultivation of adult human gingiva I. primary epithelial outgrowth from gingival explants.

The in vitro behavior of human gingiva was studied using a simple tissue culture technique.  Gingival explants from adult patients undergoing periodontal surgery were cultivated in three different media for periods up to three weeks.  It was demonstrated that primary epithelial cell cultures can be obtained under simple and predictable conditions.  The migrating cells were compact and uniform in shape and resembled the spinous cell layer of a squamous stratified epithelium.  Mitotic activity was regularly observed all over the outgrowth, which continued to grow throughout the entire experimental period.


Surface characteristics of cells from different layers of keratinized and non-keratinized oral epithelia.

Portions of clinically healthy non-keratinized and keratinized oral epithelia were removed from adult vervet monkeys (Cercopithecus aethiops).  The epithelium was separated from the underlying lamina propria by trypsin digestion, following which the epithelial cells were separated from the various epithelial layers.  These were examined with the light and scanning electron microscope.  Cells from non-keratinized epithelia have surface microplications while those from keratinized epithelia show microvilli and pits.  Two distinct types of cell are therefore present.  It is suggested that the different surface appearances are related to different types of mechanical adhesion between the cells.


The fate of vital and devitalized bone grafts in the healing of interradicular lesions.

The fate of vital and devitalized grafts of mature concellous bone from the jaws were studied in interradicular lesions in monkeys.  98 bifurcation defects were produced in premolars and molars and maintained by periodontal dressing inserted for four weeks.  Ten to 15 weeks later fresh cancellous bone was transplanted into 32 defects.  Other cancellous bone grafts were divitalized by boiling and transplanted into 34 bifurcations.  The remaining 32 defects received no grafts.  The animals were sacrificed to yield observation periods from 0 to 90 days.  After decalcification of the specimens and embedding in paraffin, serial sections were cut at 8 microns and stained.  The fate of viable and dead transplants were similar during the healing of the interradicular lesions.  Except for a few osteocytes which seemed to survive transplantation in the fresh cancellous bone, the osteocyte lacunae were devoid of cells in both types of transplants after 1 week.  The deepest located bone grafts became incorporated in new bone developed from the interradicular septum.  The more superficially located grafts were rejected or were surrounded by a cementum-like substans.  The influence of the grafts on bone regeneration was small and based solely on their osteoconductive effect.


Autoradiographic study of 3-h-proline incorporation by rat periodontal ligament, gingival connective tissue and dental pulp.

The rates of 3-H-proline incorporation by the rat periodontal ligament, the gingival connective tissue and the dental pulp were studied by autoradiography.  The rate of 3-H-proline incorporation by the periodontal ligament was 2.8 times higher than by the gingival connective tissue and 5 times higher than by the dental pulp.  These differences were significant (p smaller than 0.01).  The pattern of 3-H-proline  incorporation by the periodontal ligament was significantly different (p smaller than 0.01) from that by the gingival connective tissue and the dental pulp.  The latter two tissues exhibited a similar pattern of 3-H-proline incorporation.  The ratio of the rates of 3-H-proline incorporation by the three tissues did not correlate with the ratio of the cellular densities in the same three tissues.


Stereochemical preferences for curarimimetic neuromuscular junction blockade I: enantiomeric monoquaternary amines as probes.

Seven pairs of monoquaternary enantiomeric neuromuscular junction blocking agents were prepared in which the carbon asymmetry is adjacent to the quaternized nitrogen moiety. The tertiary amines from which the blocking species were obtained are carnegine, laudanosine, N-methylpavine, corydine, isocorydine, glaucine, and boldine. Curarimimetic potencies, obtained with an in vivo cat tongue-hypoglossal nerve preparation, were obtained for the enantiomeric methiodides of each of these amines. Possible contributions to activity be preferential binding to blood components or by selective inhibition of acetylcholinesterase also were studied. The combined studies indicate that there is a modest preference by the neuromuscular junction of the cat for monoquaternary blockers with the (s)-confirguation.


Stereochemical preferences for curarimimetic neuromuscular junction blockade II. enantiomeric bisquaternary amines as probes.

Two pairs of bisquaternary enantiomeric neuromuscular junction blocking agents as well as their diasteriomeric mesoforms were prepared in which the carbon asymmetry is adjacent to the quaternary center. The tertiary amines from which the blocking species were obtained by methyl iodide treatment were N-methylpavine and 1,1'-dodecamethylenebis(6,7-dimethoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline). Blocking potencies were determined by the mouse inclined screen assay and by the cat tongue-hypoglossal nerve technique. The mouse assay showed no statistical difference between the enantiomeric probes derived from N-methylpavine and only a modest superiority of the (R-R) isomer over the (S-S) isomer in the case of the tetrahydroisoquinoline compounds. The cat assay showed a modest statistically significant (R-R) greater than (S-S) difference in potencies in both kinds of probes. The diastereomeric meso-compounds were less active than the enantiomers in mice but were of intermediate activity in the cat determination. Acetylcholinesterase-inhibiting activity was determined for each probe to discount potency differences from this source, and no significant differences in blocking potency attributable to preferential enzyme inhibition by the probes were noted.


Sterochemical preferences for curarimimetic neuromuscular junction blockade III: enantiomeric bisquaternary amines related to benzoquinonium as probes.

Enantiomeric neuromuscular junction blocking agents, which are of the benzoquinonium type but which have a methyl group introduced adjacent to the quaternary moieties to provide an asymmetric center were synthesized and tested to determine whether the neuromuscular junction exhibits the relatively modest (R) greater than (S) superiority shown toward previously tested bisquaternaries. Testing included a mouse inclined screen assay and an in vivo cat hypoglossal nerve-tongue preparation, as well as standard estimations of anticholinesterase activity since the candidate compounds are known to have such a component in their activity spectrum. The observed 2:1 difference in blocking activity favoring the compound with an (R)-configuration is the same as that for previously tested bisquaternaries, both in direction and magnitude. Furthermore, it cannot be accounted for by preferential inhibition of acetylcholinesterase by the (S)-enantiomer. Absolute configurations of the enantiomers were assigned on the basis of comparison with compounds of known configuration.


Determination of iodochlorhydroxyquin and corticosteroids in pharmaceutical formulations.

Iodochlorhydroxyquin was separated from various corticosteroids using an acetonitrile-diatomaceous earth column. Iodochlorhydroxyquin was eluted with cyclohexane, and the corticosteroid was eluted with chloroform. Iodochlorhydroxyquin was determined by both a UV absorbance method and a new compleximetric method using the nickel chelate of iodochlorhydroxyquin. The corticosteroid was determined by the blue tetrazolium and isoniazid procedures. The average percent recovery for these four methods was 100.8, 99.4,100.7, and 99.9, respectively, for 10 known mixtures. The standard deviation for the absorbance for 10 determinations of the nickel complex was 0.002 absorbance unit (0.31%). Various characteristics of the nickel and other complexes were evaluated, including the sensitivity, solubility, and wavelength of maximum absorbance in 14 different solvents. The analyses of 23 typical products are reported, for which the standard deviation, expressed as a percentage of the amount declared, was 1.31% for the UV, 1.34% for the compleximetric, 1.49% for the blue tetrazolium, and 1.22% for the isoniazid procedures. Methods of determination in the presence of interferences are discussed.


Effect of deoxyuridine coadministration on toxicity and antitumor activity of fluorouracil and floxuridine.

The addition of deoxyuridine (UDR) to fluorouracil (FU) or floxuridine (5-fluoro-2' deoxyuridine) (FUDR) produced a substantial increase in their toxicity in BDF1 mice. Antitumor assays using sarcoma 180 tumor-bearing mice showed a concomitant increase in tumor growth inhibition for the nucleoside-drug combination over identical doses of the single drug. However, no significant increase in antitumor activity with the combination treatment was demonstrated when equitoxic doses were given. Additional support for the therapeutic equality of the single and combination drug regimens was the similarity of the therapeutic indexes for each treatment regimen involving either fluorouracil or floxuridine. The results suggested that any therapeutic benefit achieved with the combination therapy could be duplicated with either fluorouracil or floxuridine at a higher dose.