Structure-activity relationships of dimeric Catharanthus alkaloids. 2. Experimental antitumor activities of N-substituted deacetylvinblastine amide (vindesine) sulfates. While structure-activity relationships for vinblastine (VLB), vincristine, deacetyl-VLB, and deacetyl-VLB amide (vindesine, VDS) in several tumor and leukemia models have been reported previously, the present study explores these relationships for a series of N-substituted vindesine analogues. These compounds were prepared from the reaction of deacetyl-VLB acid azide with the appropriate amines and were characterized by mass spectral analysis, 1H and 13C NMR spectra, electrometric titration, and infrared spectra. N-Alkylvindesines have reduced activity compared to that of VDS against the Gardner lymphosarcoma (GLS). N-beta-Hydroxyethyl-VDS surpasses vindesine in its activity against the Ridgway osteogenic sarcoma and the GLS, whereas against the B16 melanoma it is less active than VDS. N-beta-(4-Hydroxyphenethyl)-VDS, envisaged as a substrate for the enzyme tryosinase, was shown to be more active than VDS against the B16 melanoma but has only marginal activity against the GLS. In terms of collective antitumor activity against the model systems used, vindesine emerges as the congener with optimum qualities. Bis(N-ethylidenevindesine) disulfide, the first example of a bridged bisvindesine and comparable to VDS in its antitumor profile, shows evidence of activity against a P388/VCR leukemia strain known to be resistant to maytansine as well as to vincristine.


Antianaphylactic agents. 1. 2-(Acylamino)oxazoles. The synthesis and biological properties of 35 2-(acylamino)oxazoles are described. The majority of the compounds inhibit the release of slow-reacting substance of anaphylaxis (SRS-A) in vitro from sensitized guinea pig chopped lung. In addition, several of the compounds inhibited the release of SRS-A from passively sensitized human chopped lung and protected guinea pigs from the effects of anaphylaxis in a modified Herxheimer test.


Pharmacologically active sulfoximides: 5-hexyl-7-)S-methylsulfonimidoly) xanthone-2-carboxylic acid, a potent antiallergic agent. The antiallergic activity of some xanthone derivatives containing a sulfoximide substitutent has been investigated. While 2-(S-methylsulfonimidoyl)xanthone itself was found to be inactive, a series of 7-(S-methylsulfonimidoly)-xanthone-2-carboxylic acids showed good levels of activity in the passive cutaneous anaphylaxis screen. N-Substituted sulfoximide derivatives were, without exception, less active than the corresponding unsubstituted compounds. The activity of the 7-(S-methylsulfonimidoyl)xanthone-2-carboxylic acids could be enhanced by the introduction of an alkyl or alkoxy substituent at C-5. As a result of these studies, 5-hexyl-7(S-methylsulfonimidoyl)xanthone-2-carboxylic acid has been selected for further investigation as an antipasthmatic agent.


Serotonin receptor binding affinities of tryptamine analogues. Using a rat fundus model, the serotonin (5-HT) receptor binding affinities of 27 tryptamine analogues were determined. Factors which might affect affinity were examined, e.g., lipids solubility, as reflected by partition coefficient, and pKa. Structure-activity relationships were developed and are discussed in terms of substituents on the terminal amine, the side chain, and the indole 1 position, the 5 position, and at other positions on the indolic nucleus. If lipid solubility and metabolism can be accounted for, there appears to be a parallelism between 5-HT receptor binding affinities and the hallucinogenic (psychotomimetic) potencies of several of these compounds.


Potential organ- or tumor-imaging agents. 18. Radioiodinated diamines and bisquaternaries. The purpose of this research was to employ diamines and their quaternary derivatives as carrier molecules for gamma-emitting radiation. The diamine putrescine is widespread in nature and has been reported to selectively concentrate in the rat ventral prostate and pancreas. This study confirms the selective uptake of radioactivity in the rat ventral prostate, but not in the pancreas, following administration of [14C]putrescine. The radioiodinated analogues of putrescine showed no predilection for either of these organs. On the other hand, radioactivity associated with a radioidinated quaternary derivative (3) was found to accumulate incartilaginous tissues such as trachea, intervertebral disks, and chondrosarcoma tumor in a manner simular to hexamethonium.


A new chemical series active against African trypanosomes: benzyltriphenylphosphonium salts. Antitrypanosomal activity for benzyltriphenylphosphonium salts is reported for the first time. Testing was conducted using Trypanosoma rhodesiense infected mice. Of 70 phosphorus-containing compounds tested, 21 were active. Sixteen of these active chemical species were benzyltriphenylphosphonium salts. Four were nonbenzyl triphenyl compounds. The remaining active drug was a benzyldiphenylphosphonium salt.


Synthesis and pharmacological activity of 2,3,4,5,-tetrahydro-1,5-methano-1H-3-benzazepines. 2,3,4,5-Tetrahydro-1,5-methano-1H-3benzazepine (4) has been synthesized from 2,3-dioxobenzonorbornene. Oxidative cleavage of the diketone to cis-1,3-indandicarboxylic acid, followed by closure to the corresponding amhydride, conversion to the imide, and lithium aluminum hydride reduction, gave 4. Compound 4 and its N-derivatives show no analgesic activity in the mouse hot-plate assay and little antagonist activity in a tail-flick assay.


Resolution and absolute configuration of trans-2-(2,5-dimethoxy-4-methylphenyl)cyclopropylamine, a potent hallucinogen analogue. An hallucinogen analogue, trans-2-(2,5-dimethoxy-4-methylphenyl)cyclopropylamine (DMCPA), was resolved into ints two enantiomers by fractional crystallization of salts with d- or l-O,O-dibenzoyltartaric acid. A comparison of the ORD and CD curves of the N-5-bromosalicylidene derivatives of trans-2-phenylcyclopropylamine of known absolute configuration and of the title compound established the stereochemistry of the latter to be (1R,2S)-(-) and (1s,2r)-(+). We have earlier shown that the (-) isomer shows selective behavioral effects in cats and mice. In present study it was found that the (-) isomer selectively elicits rabbit hyperthermia when compared with the (+) isomer. In view of the stereoselective ability of the (-) isomer to elicit hallucinogen-like behavioral profiles in these animal models, the proof of absolute configuration lends further support to a new model which interrelates the active binding, conformation of phenethylamine hallucinogens to that of serotonin and tryptamines.


Simultaneous solubilization of steroid hormones II: androgens and estrogens. The simultaneous solubilization of some androgens and estrogens in aqueous polysorbate 40, tetradecyltrimethylammonium bromide, and sodium lauryl sulfate was studied. The solubilizations of estradiol and testosterone were independent of each other in all three association colloids. However, if the estrogen component was ethinyl estradiol, the solubilization was dependent on the addition order. The estrogen precipitates more readily than testosterone in polysorbate 40 and tetradecyltrimethylammonium bromide, but the opposite is true in sodium lauryl sulfate. The simultaneous solubilizations of methyltestosterone or ethisterone with the estrogens tested were different from those of testosterone. The solubilization behavior of the steroids is discussed, starting with the pseudophase model and different solubilization loci. Results indicated that the free energy change of micellar binding, delta Gb, decreases with increased steroid polarity. The simultaneous solubilization cannot be predicted by delta Gb but may be explained by differences in the solubilization mechanism.


Dynamic dialysis in drug-erythrocyte interactions. Application of dynamic dialysis to drug binding to erythrocytes was found to be more complex than previously described. Drug molecules located intracellularly in erythrocytes must diffuse through erythrocytes membranes before diffusing through the dialysis membrane to the external sink in dynamic dialysis. One must consider drug within the dialysis sack and within the erythrocyte and their corresponding volumes. When binding occurs in these internal compartments, different equations result for computing Dt, the total drug within the dialysis bag. With these equations and theoretically generated data, the results of "double dialysis" on the data produced in dynamic dialysis studies of drug-erythrocyte interactions were examined. The intracellular binding of drug by the erythrocyte to a single class of preexisting noninteracting sites could be misinterpreted as cooperative binding, or binding to at least two classes of sites, when the data of dynamic dialysis are treated in the Scatchard format.


Simple and specific electron-capture GLC assay for plasma and urine ephedrine concentrations following single doses. An electron-capture GLC procedure for determination of plasma ephedrine concentrations is described. The procedure is capable of determining 2 ng/ml of ephedrine and is adequate for following profiles after 25-mg single doses. Pentane extraction of the drug and the internal standard and formation of the N-pentafluorobenzoyl derivatives were followed by GLC. Plasma ephedrine concentrations following a 24-mg dose of ephedrine hydrochloride to a human volunteer are presented. Formation of N-trifluoroacetyl, N-pentafluoropropionyl, N-heptafluorobutryl, and N-pentafluorobenzoyl derivatives and their GLC-mass spectrometric identification are discussed together with comparative electron-capture sensitivities of these derivatives toward a nickel-63 detector. The detection of the N-pentafluorobenzoyl derivative of ephedrine is at least 100-fold greater in sensitivity than detection of the N-trifluoroacetyl derivative.


Fusion of disubstituted benzenes.

Fusion of disubstituted benzenes. The entropy of fusion of 84 disubstituted benzenes was essentially constant and independent of the participation of the compounds in intramolecular or intermolecular hydrogen bonding. It was also independent of the shapes, sizes, and dipole moments of the rigid molecules studied. While the entropy of fusion was independent of these parameters, the melting point and the heat of fusion showed a direct dependence on molecular properties.


Use of nitrogen-specific detector for GLC determination of plasma bucainide. A sensitive and specific GLC determination of plasma bucainide was developed using a nitrogen-specific detector. The method permits the determination of bucainide as low as 5.0 ng/ml of plasma and provides sufficient sensitivity and reproducibility for clinical use.


Rotating-disk method for determining cutaneous metabolism. Since design and evaluation of topical dosage forms should account for both skin permeation and cutaneous metabolism, an in vitro system was formulated to determine the metabolic component of viable guinea pig skin utilizing the diffusion layer property of the rotating disk. The drug investigated was vidarabine, an antiviral agent, which was rapidly metabolized to 9-beta-D-arabinofuranosylhypoxanthine. The aqueous diffusion coefficient of the drug was determined by the capillary cell method. The rotating-disk system was standardized using benzoic acid. The dorsal skin of a guinea pig was removed after shaving, and the epidermal section was excised by a keratome. After the section was mounted on the stainless steel disk with a tissue adhesive, the preparation was immersed in a 10-ml beaker containing 5 ml of drug solution at 37 degrees. At suitable intervals, samples were withdrawn, separated by TLC, and assayed by liquid scintillation. The enzyme rate constant was 1.54 x 10(-1) sec-1.


GLC assay of verapamil in plasma: identification of fluorescent metabolites after oral drug administration. The fluorometric assay for verapamil in plasma is not useful after oral drug administration because of interference by inactive, but fluorescent, metabolites extracted along with the parent drug. A GLC method using a flame-ionization detector after silylation allows the separation of unchanged active verapamil from the metabolites and quantitation to a sensitivity of 0.025 microgram/ml. After a single oral dose of drug in dogs, up to 80% of "fluorescent verapamil" represented inactive metabolites.


Stability-indicating high-performance liquid chromatographic determination of chlorpropamide, tolbutamide, and their respective sulfonamide degradates. A quantitative high-performance liquid chromatographic method for the determination of chlorpropamide, tolbutamide, and their respective hydrolysis products, p-chlorobenzenesulfonamide and p-toluenesulfonamide, in solid dosage forms was developed. The method is stability indicating and can be used to determine the sulfonamide hydrolysis product and the intact drug in the presence of minor degradates. Method reproducibility, demonstrated by repeated injections of a calibration standard, was 1.21%. The lower limit of quantitation of the hydrolysis products, p-chlorobenzenesulfonamide and p-toluenesulfonamide, was 0.2 microgram/5-microliter injection. The accuracy of the method for intact drugs was determined by comparison of the HPLC results to those obtained by the appropriate USP or BP assays. The mean of the results obtained by the two methods differed by 0.7% for chlorpropamide and 0.3% for tolbutamide. Pure drug samples were spiked with amounts of the hydrolysis products ranging from 20 to 120% of the intact content. The mean percent recovery for p-chlorobenzenesulfonamide was 98.6%; for p-toluenesulfonamide, it was 100.6%. A qualitative TLC procedure for the detection of chlorpropamide, p-chlorobenzenesulfonamide, dipropylurea, propylurea, n-propylamine, tolbutamide, p-toluenesulfonamide, dibutylurea, butylurea, and n-butylamine is also described.


Enthalpy of bile salt-lecithin mixed micelle formation. The enthalpies for the dissolution of lecithin by sodium salts of cholic, deoxycholic, and chenodeoxycholic acids and their glycine and taurine conjugates are reported. Exothermic enthalpies were found in each case. It is suggested that heat evolution is due to a bile salt-lecithin interaction other than hydrophobic interactions. These results provide strong support for the "mixed disk" model for the complex lecithin-bile salt micelle, which requires that a substantial fraction of the bile salt molecules be incorporated within a lecithin bilayer where hydrogenbonded pair formation can occur. Calorimetric studies of the interaction between sodium cholate and nonionic, cationic, and anionic detergents yielded exothermic heats. These results suggest that these bile salt molecules partition into the detergent micelle interior as hydrogenbonded pairs.


Heterocyclic analogs of amphetamine: Thioureas, dithiocarbamates, and negatively substituted amides. A series of heterocyclic analogs of amphetamine was synthesized. The heterocycles employed included the 2-furyl, 2-thienyl, 3-methyl-2-thienyl, 3-pyridyl, and 6-methyl-2-pyridyl rings. The aliphatic amine group was converted to the N-methylthiourea, dithiocarbamate, methanesulfonyl, trifluoromethanesulfonyl, and trifluoracetyl functions since similar conversions of the beta-phenethylamine structure had shown blood pressure-lowering effects and some loss of behavioral effects. p-Chlorophenyl and 1-naphthyl analogs were also converted to these derivatives. Behavioral and other biological effects, including antiarthritic, passive cutaneous anaphylactic, and antimicrobial, were observed. The 3-methyl-2-thienyl analog of amphetamine significantly increased papillary muscle contractile force without producing arhythmias.


Percutaneous absorption of butylparaben through guinea pig skin in vitro. Percutaneous absorption of aqueous butylparaben through guinea pig dorsal skin was studied using a diffusion chamber. Polysorbate 80 increased the solubilized concentration but decreased penetration of the preservative. Polyethylene glycol 400 also reduced the amount of penetration. Propylene glycol was less effective than polyethylene glycol 400. Preservative activities of these systems on several microorganisms were evaluated on agar plates. The relationship between the butylparaben penetration and preservative activity is discussed.


High-performance liquid chromatographic assay for the antiprotozoal agent, tinidazole, in human plasma. A rapid and specific high-performance liquid chromatographic (HPLC) assay for the quantitative determination of tinidazole in human plasma is described. After a simple extraction procedure, the compound is analyzed by HPLC using a reversed-phase column and a UV detector. Quantitation is accomplished using an external standard, and peak areas are determined with an integrating computer. The average recovery of tinidazole over a concentration range of 0.20-20.0 microgram/ml was 86.9 +/- 2.8% SD. The maximum sensitivity of the assay is approximately 0.10 microgram/ml; it is suitable for use in pharmacokinetic studies following administration of therapeutic levels of tinidazole to humans.


Thermal characterization of citric acid solid dispersions with benzoic acid and phenobarbital. The glass transition temperatures of citric temperatures of citric acid glass were determined by differential scanning calorimetry to be 10.2 and 13.5 degrees for in situ and bulk-prepared samples, respectively. Mechanical stress on citric acid glass induced foci for crystallization. Benzoic acid addition to citric acid glass decreased its glass transition temperature while phenobarbital addition increased its glass transition temperature, the latter forming a glass solution.


Paired-ion reversed-phase high-pressure liquid chromatographic assay of pentobarbital-pyrilamine suppositories. The assay of suppositories containing pentobarbital and/or pyrilamine in a water-soluble polyethylene glycol base by high-pressure liquid chromatography is described. No extraction is required. The suppository is dissolved in the mobile phase. This solution is diluted with an internal standard stock solution containing phenobarbital. Chromatographic conditions include a C18 bonded microporous silica column and a mobile phase of 65% 4 x 10(-3) M n-butyl sodium sulfonate in 1% acetic acid and 35% acetonitrile. The procedure using commercial products gave results comparable to those obtained by GLC.


Chemical reactions in cephalosporin allergy: high-pressure liquid chromatographic analysis of cephalosporin aminolysis kinetics. Cephalosporin reaction with protein amino groups is fundamental to cephalosporin allergy. Cephalothin and cefazolin reaction kinetics with epsilon-aminocaproic acid, B-alanine, and glycine in aqueous solution were investigated. All reactions were conducted at 35 degrees and 0.5 ionic strength and were followed by ion-exchange high-pressure liquid chromatography. The aminolysis rate constants can be expressed as terms representing uncatlyzed or water-catalyzed amine reaction, self-assited nucleophilic reaction, and hydroxide-ion-catalayzed nucleophilic amine attack on the beta-lactam moiety. Cephalothin and cefazolin react with amines as readily as penicillin G. The UV spectra of several cephalothin-glycine reaction products were recorded, and their possible structures are discussed.


Hydrocortisone stability in human feces. Hydrocortisone stability in human feces was studied under various conditions to determine whether stability accounts for the variable effects of hydrocortisone enemas. Recovery from feces and assay specificity were assured using dual isotopes, TLC separation, and liquid scintillation counting. Hydrocortisone degraded slightly from 7 to 26% in 24 hr when incubated in fresh human feces at 37 degrees. Less than 7% degradation occurred in feces stored at 10 degrees, and negligible degradation occurred with hydrocortisone in water at 37 degrees. Fecal bacteria may account for the observed degradation. Hydrocortisone stability in feces may contribute to local persistence and may account partly for its efficacy in ulcerative colitis treatment.


Plasma pentazocine radioimmunoassay.

Plasma pentazocine radioimmunoassay. A sensitive and specific radioimmunoassay of dog and human plasma pentazocine is described. Rabbit antiserum and the second antibody method separated bound from free pentazocine. The radioimmunoassay employed an 125I-labeled radioligand and required extraction from the sample prior to quantitation. The method had a detection limit of approximately 200 pg/assay tube (1 ng/ml). The assay was used successfully to measure pentazocine in the plasma of beagle hounds given 0.3 mg of pentazocine/kg iv. The decline in plasma levels fitted a two-compartment body model with a 100-min mean overall half-life and a 3.2-liters/hr mean plasma clearance rate.


Drug entrapment for controlled release in radiation-polymerized beads. The preparation of beads including polymer, vinyl monomer, and drug was carried out by low temperature, radiation-induced polymerization. Complete spherical particles were obtained when ethanol was the precipitation medium Polymers such as polymethyl methacrylate and polystyrene were dissolved in various glass-forming monomers, and the drug was dispersed in the mixture. The mixture was dropped into cold precipitation media. The formed monomeric particle was irradiated at low temperatures to produce polymerization. The drug release profiles from polymerized particles were changed by varying the glass-forming monomer and the precipitation medium.


UV studies of nucleic acid vase complexation with isoproterenol in different solvents. Charge transfer complex formation between nucleic acid bases and isoproterenol was demonstrated from UV absorption measurements. The solvent polarity effects on equilibrium constants were investigated. The solvent systems containing 0.1 N HCl were 20% aqueous ethanol, water, and water containing sodium chloride. The equilibrium constants, calculated from UV absorption data by the application of the Benesi-Hildebrand equation, were small and increased with increasing ionic strength. Equilibrium constant wavelength dependence was demonstrated in some cases.


In vitro and in vivo availability of hydrophilized phenytoin from capsules. The effect of phenytoin hydrophilization on the liquid penetration rate into prepared plugs, on the disintegration time, on the in vitro release rate, and on in vivo absorption in humans was studied. Hydrophilization was performed by intensive mixing of the hydrophobic drug with a small amount of methylcellulose solution. Liquid penetration into the treated plugs was independent of the liquid wetting potency and extremely high compared to the pure drug plugs. Analogous results were obtained for the disintegration time and in vitro release rates from capsules loaded with pure and treated drug. A bioavailability study in seven healthy volunteers showed immediate absorption of the treated drug but a 1-hr absorption lag time for the pure drug.


Oxybutynin influence on autonomic measures in dogs. In the conscious dog, the most apparent oxybutynin effect was a dose-related tachycardia. Associated with this heart rate increase were a very slight, sometimes significant, elevation in diastolic pressure and an insignificant increase in systolic pressure. Under pentobarbital anesthesia, the systolic/diastolic arterial pressure oxybutynin responses were reversed and showed a dose-related systolic and diastolic hypotension. However, the tachycardic response to oxybutynin still appeared. The arterial pressure and heart rate responses produced by the autonomic agents were altered by the oxybutynin treatment in a pattern indicative of an anticholinergic mechanism of action. Differences in many response profiles were observed with either conscious or anesthetized dogs, but the statistically significant inhibition of the acetylcholine-induced systolic and diastolic arterial pressure and bradycardic responses were constant in both conditions. Oxybutynin is an anticholinergic agent with mild to moderate cardiovascular activity.


Amino acid effect on aspirin stability in propylene glycol. Temperature stability studies were conducted on 0.36 M (6.5% W/V) aspirin solutions including either 0.02 M L-methionine or 0.02 M histidine in propylene glycol. Aspirin was determined spectrophoto-fluorometrically as salicylic acid content at 412 nm. A 0.36 M aspirin in polyethylene glycol 400 solution was studied concurrently. Aspirin degradation rate constants, k, obtained from semilogarithmic plots of percent drug remaining versus time at 30-70 +/- 0.5 degrees were used for preparing Arrhenius plots. Good correlation was seen between predicted aspirin stability and experimental k25 degrees values. L-Methionine and histidine markedly reduced aspirin stability.


Radiochemical plasma salicylamide assay using ring-labeled tritiated salicylamide. A rat plasma salicylamide assay was developed using ring-labeled tritiated salicylamide, synthesized by reacting salicylamide with tritium oxide in the presence of heptafluorobutyric acid. The reaction yielded 3H-salicylamide of specific activity up to 8.41 mCi/mmole, 60% yield. Plasma containing 3H-salicylamide and its metabolites was extracted with a toluene-based scintillation fluid, which was subsequently counted. Specificity for free salicylamide was demonstrated by radio chemical and standard fluorescence plasma salicylamide level-time curves. Specificity resulted from nonextraction of the salicylamide sulfate and glucuronide metabolites. Sulfatase and beta-glucuronidase treatment allowed the analysis of plasma sulfate and glucuronide conjugates as free salicylamide. This procedure should be effective for the analysis of salicylamide and its metabolites in the presence of similar phenolic compounds.


High steady-state levels of uric acid produced in rats by dietary training and potassium oxonate. To reduce the inherent variability in serum uric acid levels of animals allowed ad libitum exposure to food containing potassium oxonate and uric acid, male Sprague-Dawley rats were trained to eat their daily food allotment in a 1.25-hr period each morning. After training the rats were fed a food mixture containing 5% potassium oxonate and 2% uric acid (w/w each). Serum blood levels of uric acid reached a steady state within 2 hr; these levels were maintained for an additional 4 hr. It is believed that the stomach emptying rate is a zero-order process under these experimental conditions.


High-performance liquid chromatographic determination of cephacetrile. A rapid and accurate quantitative determination of cephacetrile in finished bulk and dosage forms is reported. The high-performance liquid chromatographic method is free of interference by acetyl hydrolysis products and synthesis by-products. The assay can be performed in about 15 min, affording less than 0.7% coefficients of variation within and between days. The chromatographic results are in good agreement with the microbiological assay requested by the "Code of Federal Regulations" for certification of cephacetrile sodium.


Preparation of semisynthetic (+)-tubocurarine chloride. Semisynthetic (+)-tubocurarine chloride (II) was prepared by monoquaternization of (+)-tubocurine. The method involved treating (+)-tubocurine with a 0.5 M equivalent of hydrochloric acid prior to quaternization with methyl iodide, followed by neutralization and iodide-chloride ion-exchange. Column chromatography and crystallization procedures were utilized for pure semisynthetic II preparation. The neuromuscular junction blocking activities of the semisynthetic and commercial II were determined by the in vivo cat hypoglossal nerve-tongue muscle preparation. No delectable differences among physical constants, spectral data, and neuromuscular junction blocking activities were noted between the commercial product and the semisynthetic II. This result substantiates the chemical and biological data for the well-accepted new formula for II. The unexplained M + n14 mass spectral peaks shown by the curare-type bases are characteristic of the molecular species rather than a result of contaminants.


Simultaneous determination of isosorbide dinitrate and its mononitrates in human plasma by capillary column GLC. A previously described electron-capture GLC method for determination of isosorbide dinitrate in human plasma was adapted for the simultaneous determination of isosorbide dinitrate, isosorbide 2-mononitrate, and isosorbide 5-mononitrate using a capillary column. Quantitation was done with two internal standards. The lower limits of detection were approximately 0.5 ng/ml of plasma for isosorbide dinitrate, 2 ng/ml for isosorbide 2-mononitrate, and 20 ng/ml for isosorbide 5-mononitrate.


Improved method for morphine extraction from biological samples. Methadone morphine, or naloxone extraction from brain homogenates, plasma, and urine is described. An aqueous sample was loaded on a surgical gauze support, which was washed with extracting solvents. Aqueous samples remained on the support, and nonpolar drugs partitioned into the lipophilic extracting solvent. The procedure recovered 80-100% of nanogram levels of methadone, morphine, or naloxone from biological samples. In addition, an approximate 10-fold timesaving capacity was demonstrated compared to standard liquid-liquid extraction techniques.


Effect of water-soluble carriers on morphine sulfate release from a silicone polymer. The influence of gelatin, sodium lauryl sulfate, lactose, and sodium alginate on morphine sulfate diffusion from cylindrical silicone polymer pellets was examined in isotonic pH 7.4 phosphate buffer. These water-soluble carriers caused the pellets to swell in aqueous media. Sodium alginate exerted the greatest influence on drug release. The morphine sulfate diffusion rate from the cylindrical pellets increased as the matrix alginate content increased up to 20%. Water-soluble carrier incorporation into silicone polymeric matrixes permits controlled release of water-soluble drugs that otherwise would be released extremely slowly from the polymer. Drug diffusion from the silicone matrix containing sodium alginate followed second-order kinetics. The release mechanism probably involves the creation of pores or pathways through the matrix secondary to the swelling.


Wikstromol, antitumor lignan from Wikstroemia foetida var. oahuensis Gray and Wikstroemia uva-ursi Gray (Thymelaeaceae). The ethanol extracts of Wikstroemia foetida var. oahuensis and Wikstroemia uva-ursi showed antitumor activity against the P-388 lymphocytic leukemia (3PS) test system One PS-active constituent of both plants was the lignan wikstromol. Several inactive compounds were identified as daphnoretin, pinoresinol, and syringaresinol.


Paradoxical traps in therapeutics: some dilemmas in medical ethics. The doctor-patient relationship is examined an emphasis on the comparison between professional and moral principles. Many therapeutic measures have opposite-directed alternative steps with an equal degree of justification, so that no logical preference is attainable and conflicts ensue. Thus patients come for relief and are ordered to endure further pain and discomfort; or weaker individuals exaggerate their complaints hypochomdriacally, and thus need a great deal of understanding, yet paradoxically they are prone to receive less support than stronger ones. Further conflicts arise between our devotion to human well-being and dignity, and our obligation to disrespect some of their rights for self-determination. Furthermore, various dutifully performed doctoring activities run counter to our own social needs and interests; last, but not least, human imperfection colours some of our decisions, putting a definite blemish on their value. In conclusion, physicians must bear the constant burden of paradoxically-opposed alternatives, and they confront pitfalls of worongdoing at every therapeutic step. Their only guidelines are intuition and professional dedication.


When consent is unbearable: an alternative case analysis. Dr Agich takes up a previous difficult case related by Dr Kottow in an earlier issue of the Journal. He analyses the three ethical problems as presented in the case and offers his own opinion of it as well as his own conclusions with regard to the medical ethical aspects of it. Unlike Dr Kottow, Dr Agich's reading of the case indicates that the application of the principle of informed consent does not rule out ethical decisions for the physician, but emphasizes the relevance of ethical analysis beyond the issue of informed consent.


Help yourself to good health?

Help yourself to good health? In recent years more support has been given to the idea of Health Education and there are strong reasons for believing that such schemes of prevention may receive much more financial backing from governments. It is being realised, also, that many of our 'ills' may be attributed to an over-reliance on medical technology. There is reason to believe that in years to come the emphasis will be placed on the fostering of mental health in programmes of Health Education enabling the individual to take a responsible attitude in learning how to seek out health hazards for himself without over-reliance on others.


A follow-up neurobiological study: why volunteer? There is usually great concern over the use of psychiatric patients for clinical research, as it raises the ethical and legal issues of human dignity and autonomy. In this paper the authors describe and evaluate a follow-up neurobiological study of patients who had been discharged from a psychiatric research ward at least ten months earlier. It is pointed out that such studies are rare and that the writers were provided with the unique opportunity to examine attitudinal and motivational dimensions involved in the patients' agreement to participate in the study.


Regulation, professional responsibility, and market forces in the health care field. In this paper, the author first deals briefly with the general interrelationship of market forces, government regulation, and professional responsibility. He then reviews the Federal Trace Commission's ongoing program involving competition in the health care field. Finally, he attempts to cover some prevalent concerns and what he believes are some misconceptions in the medical community about the commission's involvement in the health area.


Departmental review in medical schools: focus and functions. Approximately two-thirds of all U.S. and Canadian medical schools have provisions for the evaluation of their departments on a periodic or ad hoc basis. Most of these institutions have initiated the departmental review process since 1970. It appears that use of the practice is increasing and is becoming an important tool in providing guidance for departmental activities, programs, and leadership. Departmental review may be conducted in a variety of ways with varying levels of intensity, flexibility, and skill, depending on the environment, resources, dean, department chairmen, faculty, and attitudes of university administrators. Departmental review may be employed to identify, elaborate, document, and/or contribute to the resolution of current and potential organizational concerns. When carefully administered, the process can be a stabilizing rather than a disruptive force; however, by itself, it does not solve major institutional problems.


The costs of an institutional review board. The Institutional Review Board (IRB) at the University of Texas Health Science Center at San Antonio is charged with the responsibility of review of all protocols involving human subjects in research. A large medical institution has many clinical protocols requiring review. The review of hundreds of protocols per year requires much time from the IRB staff, the committees which review the protocols, and the investigators who prepare them. This cost is borne by the institution. Additional federal regulations add to the complexity of review and require additional time on the part of all concerned. An attempt has been made to document the costs of operation of the IRB in a single group of institutions. About 850 new or renewal applications are reviewed each year for $100,000 or about $100 per application. This is a sizable financial burden to be placed on the average medical school. Legislation is now proposed to support IRB activities by direct grants from the federal government.


The regional health education center: Illinois AHEC as an organizational prototype. Illinois is one of 11 states to develop area health education center (AHEC) projects as recommended by the Carnegie Commission on Higher Education and funded by the federal government. The experience of the University of Illinois Medical Center with its project suggests the evolution of a regional health education center concept in which AHEC functions are undertaken within the context of regional schools of medicine. A conceptual model is presented here which relates to regionalized health professions education, with medical education as the locus for its development.


The premed stereotype.

The premed stereotype. For this study of attitudes surrounding premedical education, the authors examined the consequences of competitive pressure for admission to medical school. Responses of premedical and other students were compared on a number of topics including: self-descriptions, premedical stereotypes, occupational values, attitudes toward various occupations, and perceptions of college pressures. Differences are reported for premedical subgroups, including science and nonscience majors, men and women, and minority group members and others.


Postgraduate training for general practice in the United Kingdom. Although the role of general practice is well established in the United Kingdom's National Health Service, formal postgraduate training for primary care practice is a recent development. Trainees may enter three-year programs of coordinated inpatient and outpatient training or may select a series of independent posts. Programs have been developed to train general practitioners as teachers, and innovative courses have been established. Nevertheless, there is a curious emphasis on inpatient experiences, especially since British general practitioners seldom treat patients in the hospital. In their outpatient experiences trainees are provided with little variety in their instructors, practice settings, and medical problems. The demands on this already strained system will soon be increased due to recent legislation requiring postgraduate training for all new general practitioners. With a better understanding of training for primary care in the National Health Service, those planning American primary care training may avoid the problems and incorporate the attributes of British training for general practice.


Medical student clinics: the basis for a clerkship in family practice. In this paper, the authors describe a popular elective clerkship in family practice a Community Hospital, Santa Rosa, California. The key element in the clerkship for the past six years has been the student's General Medical Clinic. Here, under the supervision of a family physician teacher, senior medical students care for patients with a wide variety of problems. The students work as a team with family nurse practitioners. The clinic has succeeded in offering patients an alternative to the emergency room and an entrance to the hospital's Model Family Practice Center. The students give the clerkship a very high rating as an educational experience. It has also been viable economically.


Identification with the patient in a psychiatry clerkship. In an earlier study medical students were found to view psychiatry with considerable anxiety partly because of an underlying fear of contact with psychiatrists and mentally ill patients. After supervising medical students for several years, the present authors found that the students' reactions to psychiatric patients may include, besides fear, a variety of behaviors which seem to reflect a process of identification with the patient. In this paper they describe various patterns of this process and their effects upon student-patient interaction.


The effects of the polyene antibiotic mepartricin on polymorphonuclear leucocyte function: an in-vitro study. Mepartricin, a polyene antibiotic with candidacidal and trichomonicidal activity, was found to be without toxic effects for human polymorphonuclear leucocytes; the drug seems to be unable to enter the human cells. Some synergism between the antifungal activities of mepartricin and of human leucocytes is seen if Candida cells are pre-incubated with sub-lethal concentrations of the drug.


Relationship between antibiotic resistance, the production of "virulence factors", and virulence for experimental animals in Staphylococcus aureus. Variants that had lost some of their antibiotic-resistance determinants were selected from a multiple-antibiotic-resistant strain of Staphylococcus aureus. When tested by subcutaneous injection into guinea-pigs, and measured as the number of cocci needed to produce a skin lesion of an arbitrarily chosen diameter, the virulence of strains fell progressively with loss of resistance determinants. When the staphylococci were injected intracutaneously into mice, however, the results were less easy to interpret, but loss of resistance appeared to be associated with a reduction of the slope of the dose-response line. There was no association between the antibiogram of the strains and their production of certain enzymes and haemolysins.


Evaluation of the lysostaphin-susceptibility test for the classification of staphylococci. A collection of 39 staphylococcal strains of known peptidoglycan structure was cultivated on media supplemented with different peptones or with glycine or inosine. The susceptibility to lysostaphin 50 mg/litre was then checked by a plate-dilution method. The results of the test were dependent on the conditions of growth. Another collection of 403 staphylococcal strains from international collections, clinical material and healthy human skin was also tested for susceptibility to lysostaphin. No uniform pattern of the lysostaphin sensitivity in individual staphylococcal species was found. The lysostaphin-susceptibility test thus seems to be of little value in the classification of staphylococci for the purposes of medical microbiology.


Age-related susceptibility of mice to staphylococcal infection. The virulence of six strains of staphylococci in experimental subcutaneous infection in mice of three age groups (3, 10 and 21 days) was studied. The results showed an age-related susceptibility to infection, in that the newborn mice were more susceptible than older mice to death and lesion formation. Resistance, i.e., ability to survive challenge of 10(7) c.f.u., developed at about 5 days. The strains used varied in toxin and enzyme pattern and there were marked differences in response to challenge as measured by mortality and lesion development. The Staphylococcus epidermidis strain was least virulent, while some strains of S. aureus produced lesions at low doses (10(4) c.f.u.). Two distinct types of lesion were observed, abscesses and necrotic lesions. Development of necrotic lesions appeared to be correlated with the ability to produce toxin in vitro.


Techniques utilizing real time stereo scanning electron microscopy in the microdissection of biological tissues. The use of the scanning electron microscope (SEM) for the microdissection of biological tissue is described. The dissections were accomplished by the use of electrolytically pointed needles that were positioned in a simply constructed holder. The operations were viewed using real time stereo SEM, and recorded on videotape. Various methods for photographing the videotape recordings are outlined.


A rapid method for estimating volume ratios. A rapid stereological method is proposed for estimating the volume ratio V(K2)/V(K1) of two bodies K1, K2 such that K2 is embedded in K1. If K1, K2 fit approximately into a 'star specimen' model, then a single section, taken at a certain level through K1, provides a rather accurate estimate of V(K2,/V(K1). For a population of moderately similar star specimens, a constant sectioning level can be estimated from a sample using a least squares criterion. A pilot experiment involving mouse lymph nodes, aimed at estimating the mean and the variance of the individual fractions V (paracortex)/V(node), indicates a fair robustness against deviations from the model.


A new method for scanning electron microscopic visualization of dermal elastic fibres. Autoclaving sections of dermis for 8 h followed by fixation, dehydration and xylol-air drying yields a pure preparation of elastic fibres for scanning electron microscopy which retains the native architecture of this component. Elastic fibres were intertwined in a complex fashion with numerous branches. Fibres were predominantly cylindrical in the upper dermis and became larger and more elliptical in the deeper dermis. This method produces a means to study of organization of elastic fibres in a variety of disorders in which dermal changes are prominent.


Electron microscope autoradiography of a diffusible intracellular constituent, using freeze-dried frozen sections of mammalian intestinal epithelium. Autoradiograms were prepared from freeze-dried frozen sections of rat intestinal epithelium, previously incubated in vitro with 3H-galactose. Although the sections showed evidence of disruption by ice-crystal growth during freezing, they retained sufficient recognizable ultrastructure to facilitate the identification of several subcellular compartments. Statistical analysis of the grain distribution indicated that a significant non-uniform distribution of the labelled galactose occurs in the absorptive cells during transport, and is maintained in the sections throughout the various stages of autoradiography.


Topography and thickness of air-dried human platelets measured by correlative transmission and scanning electron microscopy. Human platelets rapidly air-dried on carbon-coated grids were examined by transmission and scanning electron microscopy. Whole cell mounts were photographed in a transmission electron microscope (TEM), coated with gold, and then examined in a scanning electron microscope (SEM). The thickness of the cytoplasm towards the centre of the cells was estimated to be 20-40 nm, and the rim of dense material surrounding the cells was 40 nm thick. Some dense bodies stood out as much as 100 nm above the dried cytoplasm. These measurements are important for evaluating cytoplasmic volume during microprobe analyses of air-dried platelet preparations.


Partial regeneration of the above-elbow amputated rat forelimb. I. Innate responses. Although a number of recent studies describe the facilitation of limb regeneration by electrical and other forms of stimulation, little is known of innate regenerative capacity in the mammalian limb. The present report describes spontaneous regenerative responses following subtotal forelimb amputation in the young white rat. In one group of animals the forelimb was amputated through the lower humerus and the skin sutured closed. In a second group, adjacent muscle tissue still attached to bone at its origin(s) was interposed between the cut surface of the humerus and the skin. Among animals of the first group (skin closure only) bone growth and limb regenerative responses were generally not observed. Animals of the second group displayed significant elaborations of cartilage and bone at the limb terminus. The appearance and subsequent modification of these tissues suggest that some capacity for limb regeneration exists innately in the young rat and can be more readily evoked than has been recognized heretofore. It is concluded that extant and forthcoming reports of electrically stimulated skeletal tissue growth, repair and regeneration among eutherial mammals should be examined to determine whether reported responses to stimulation represent advances beyond what might be expected from innate replacement processes alone.


Partial regeneration of the above-elbow amputated rat forelimb. II. Electrical and mechanical facilitation. This investigation was undertaken to examine the observations of Becker ('72) pertaining to the electrical facilitation of partial limb regenerative responses by means of Ag-Pt wire couples applied to the limb stumps of young, forelimb-amputated white rats. Additionally, in order to examine the possible role of mechanical effects of such device implantations, we have employed uncoupled devices delivering no current or potential difference. In the present experiments, in response to coupled device implantation, cartilage and bone were actively formed in the vicinity of the Pt electrode tip. These tissues contributed to the lengthwise extension of the limb and to the partial restoration of the distal humeral extremity. In limbs bearing the uncoupled electrical devices, qualitatively similar responses were noted, but osteogenesis was diminished in extent compared to that seen in limbs bearing the active or coupled devices. It is therefore necessary to consider the role of mechanical factors in the elicitation of the observed regenerative responses. Myogenesis was enhanced in electrically stimulated limbs, but not in those rats bearing uncoupled devices.


The management of laryngeal cancer.

The management of laryngeal cancer. Carcinoma of the larynx is best managed in an interdisciplinary centre with wide therapeutic and rehabilitative services. Current management is confused, but may be simplified by consideration of three groups: 1) no fixation of laryngeal structures, 2) fixation of laryngeal structures and extension beyond the larynx, 3) all others including carcinoma in situ, verrucous carcinoma, transglottic tumor, and squamous carcinoma with marked airway obstruction. The rational treatment of Group 1 glottic tumors is primary radiation, which produces 75% crude and 92% corrected five year tumor free survival. If surgery is undertaken as primary treatment or for salvage, a vertical hemilaryngectomy may preserve the voice. Group 1 supraglottic carcinomas may be divided into supraglottic and marginal. Thirty per cent have palpable nodes at diagnosis and a further 20% occult disease in the cervical chain. Irradiation of neck nodes or block dissection is an integral part of treatment. The indications for a supraglottic horizontal partial laryngectomy are outlined. Where indicated this has five year survival figures of 70%. Recurrence is usually in the neck. Marginal tumors have a 50% five year survival rate when treated by conservation surgery combined with pre-operative radiation. Recent radiotherapeutic advances have improved treatment of these lesions. Thirty to 40% of patients with Group 2 tumors have regional metastases at presentation. There are no fixed protocols for treatment of these patients, less than 50% of whom will survive five years tumor free. The difficulty in carrying out a protocol based on pre-operative radiation with planned surgery is outlined. Under optimum conditions treatment should be primary radiation with salvage surgery for failures or recurrence, for the results are almost as good as primary surgery but 30% of larynges are saved. The difficulties of diagnosing recurrent tumor in irradiated tissue are discussed. Care must be taken to recognize that group of patients in whom tissue edema is the result of perichondritis rather than tumor recurrence, because in these patients surgery is extremely hazardous. Problems of diagnosis and methods of treatment of carcinoma in situ, and verrucous carcinoma are described. Transglottic carcinomas are defined and treatment is primary total laryngectomy with appropriate neck dissection. Similarly if tumors are causing major airway obstruction, treatment is by primary laryngectomy.


Granulomatos cervical lymphadenitis.

Granulomatos cervical lymphadenitis. One hundred and thirty-nine cases of pathologically involved cervical lymph nodes have been reviewed, and granulomatous lymphadenitis found in 29 patients (20.9 per cent). There were two cases of sarcoidosis, 10 of tularemia, five patients (3.6 per cent) had tuberculous lymphadenitis and 12 (8.7 per cent) fulfilled commonly accepted criteria for infection with atypical mycobacteria. The clinical features of the latter condition, which is probably insufficiently recognized, are reviewed, and surgery is recommended as the primary treatment modality.


[Sialoscintimetry in disorders of the facial nerve]. The comparative study of submandibular gland function before and after unilateral section of the chorda tympani, is used to verify the validity of sequential scintigraphy as a method of appraising the function of the salivary glands. Sequential scintigraphy of the submandibular glands was evaluated as a new, simple, and dependable test of the function of the submandibular glands, in topographical and prognostic assessment of 10 patients with unilateral idiopathic facial paralysis. The results of this neurodiagnostic test were compared and were found to be in close agreement with the results obtained by submandibular salivary flow measurement. The use of sequential scintigraphy of the submandibular glands constitutes a valuable tool in the clinical evaluation of patients with peripheral facial nerve disorders.


A critical evaluation of the glycerol test in Ménière's disease. The glycerol test is currently widely used to evaluate Menière's disease, particularly in regard to its suitability for decompressive surgical therapy. Our previous findings have shown that patients with Menière's syndrome respond extremely well to placebo treatment. We have investigated the effect of psychological factors in the outcome of the glycerol test. It was administered twice to each of a group of patients with a typical Menière's syndrome. On one occasion the patient was told the hearing would improve, and on the other that the hearing would worsen. The same dose of glycerol was provided for each test, but with a different flavor. The sequence of test was randomly applied. The audiologist was unaware of instructions given to the patient. The patients usually responded as instructed. These results are discussed with particular regard to the risk of patient selection for surgery.


The mechanics of benign paroxysmal vertigo. Benign paroxysmal vertigo (BPV) is a disorder of the vestibular labyrinth. The clinical features can be explained by an abnormality in the posterior semicircular canal. Under the influence of gravity, a density differential between the endolymph and the cupula will cause displacement of the cupula when changes in head position occur. The presence or absence of fatiguability is a useful test as it helps define etiology, prognosis, and therapy. At the risk of adding yet another classification of nystagmus to the literature, we submit that division of BPV into two types (fatiguable and nonfatiguable) will simplify and rationalize the management of this common complaint.


Inverted papilloma and squamous cell carcinoma. Since 1960, 15 patients with histologically proven inverted papilloma of the nose and paranasal sinuses have been treated at the University Hospital, City Hospital, and V. A. Hospital in Boston. Eight of these patients have been found to have squamous cell carcinoma in the pathological specimens; four of these patients have died of carcinoma. Wide surgical excision, usually through a lateral rhinotomy with exenteration of the invaded sinus as indicated, was the therapy of choice. In spite of the published incidence of carcinoma in inverted papilloma of approximately 7-24 per cent, we have found a much higher rate. We therefore, suggest that meticulous microscopic examination of multiple sections of inverted papilloma specimens be carried out in a search for evidence of squamous cell carcinoma. Inverted papilloma should be treated as a premalignant lesion by appropriate radical surgery.


Styloid, velar, and pharyngeal muscles in cleft palate. Anatomical findings in elderly cadaver with unrepaired cleft palate. Dessection of the musculature of the palate, pharynx, and styloid process was carried out in a cadaver aged 78 years with an unrepaired complete cleft palate. A new muscle named "the accessory stylohyoid" was found to be attached to the styloid process and the lesser cornu of the hyoid bone bilaterally. Anatomical changes in other muscles especially the stylopharyngeus and palatopharyngeus are described. The functions of these muscles are discussed on the basis of the anatomical findings and previous observations of other investigators.


Nasal aerodynamics.

Nasal aerodynamics. The historical development of nasal aerodynamic measurement is discussed. Attention is drawn to the absence of a consensus on adequate rhinometric techniques and standards, which hampers the elevation of rhinometry to the universal clinical status of audiometric, impedance, or nystagmus measurement. A computer-aided technique is described which overcomes some problems of rhinometry, by measurement of the amount of energy the body devotes to moving respiratory air through each separate nasal cavity. Although the approach is different, our preliminary results confirm previous work and suggest that assessment of the work of breathing is a promising technique in the investigation of nasal respiratory airflow.


Restriction insensitivity in bacteriophage T5 I. Genetic characterization of mutants sensitive to EcoRI restriction. Unmodified bacteriophage T5 is able to grow normally on bacterial hosts carrying three different Escherichia coli restriction systems, EcoK, EcoPI, and EcoRI. Under the same conditions, the plating efficiency of bacteriophage gamma is less than 10(-9). At least in the case of EcoRI, this lack of in vivo restriction is not due to lack of restriction sites on the T5 DNA molecule. These observations suggest that bacteriophage T5 specifies one or more restriction protection systems. Mutants (ris) of T5 have been isolated which confer sensitivity to EcoRI restriction but not to EcoK or EcoPI. The mutations are located in the pre-early region of the genetic map but are too far apart to be alleles of a single gene. Complementation studies show that the ris mutants can be helped to grow on the EcoRI-restricting host by coinfection with T5+. This result provides evidence for a restriction protection function but does not necessarily show that the ris mutants are defective in such a system.


Mitochondrial calcium uptake during infection of chicken embryo cells with Semliki Forest virus. The key role of the mitochondria in the regulation of cellular Ca2+ led to a study of mitochondrial Ca2+ uptake during the infection of chicken embryo cells with Semliki Forest virus. Mitochondrial Ca2+ uptake was stimulated during the first 5 h of infection but declined later in infection. The early stimulation suggests an increase of cytoplasmic ionized Ca2+, whereas the later decrease indicates mitochondrial injury. This functional deterioration was correlated with an increase of the permeability of the inner mitochondrial membrane. Polarographic experiments showed that electron transport is impaired, whereas transduction of energy to Ca2+ uptake is intact.


Restriction insensitivity in bacteriophage T5. II. Lack of EcoRI modification in T5+ and T5ris mutants. Neither bacteriophage T5+ nor its EcoRI-sensitive ris mutants became modified during growth on an EcoRI-modifying host. For this reason, the rare ris plaques able to grow on the EcoRI-modifying host were always due to revertant phage rather than to modified ris mutants. The ris mutations resulted in the creation of new EcoRI cleavage sites in the terminally repetitious first-step transfer DNA, and analysis of T5 ris revertants showed loss of these sites and restoration of the wild-type restriction pattern. Natural EcoRI sites present in the second-step transfer DNA were never lost in T5ris revertants, indicating that these are irrelevant to in vivo restriction and are protected during growth on the restricting host.


Mapping of class II promoter sites utilized in vitro by T7-specific RNA polymerase on bacteriophage T7 DNA. Restriction endonuclease Bgl II cleaves T7 DNA at a unique site (28.76% on the standard T7 map), yielding two fragments of molecular weights 18.9 x 10(6) (A) and 7.6 x 10(6) (B). Fragment B, representing the leftmost portion of the genome, has been purified by zone sedimentation. Transcription of fragment B by T7-specific RNA polymerase gives only r-strand-specific RNA. Analysis of the products by polyacrylamide gel electrophoresis reveals four major RNA species which have apparent molecular weights of 2.1 x 10(6), 1.36 x 10(6), 0.85 x 10(6) and 0.125 x 10(6), respectively. Each of these RNAs is reduced in size when transcription is carried out with fragment B, which has been shortened by treatment with Escherichia coli exonuclease III. Therefore, each of the transcripts must be terminated at the right end of fragment B. Analysis of the molecular weights of the four transcripts produced from whole and from exonucleolytically shortened fragment B suggests that these transcripts are read from promoters located at 13.5, 18.9, 22.6, and 27.9%, respectively, on the standard T7 map. Hence, there are at least four promoters governing the transcription of the class II region. Transcripts initiated at these promoters on intact T7 DNA appear to read through the class II and part of the class III genetic region and terminate at the strong terminator for T7-specific RNA polymerase near 61%. Transcription of fragment B which has been cleaved with the restriction endonuclease Hpa I seems to activate a fifth promoter for T7-specific RNA polymerase. This promoter appears to be identical to the promoter previously described by Oakley and Coleman (Proc. Natl. Acad. Sci. U.S.A. 74:4266-4270, 1977) that maps near 15% on the standard T7 map. Little or no RNA is read from T7 Bgl II fragment B, which has a mobility expected for a transcript read from this promoter. However, upon cleavage with Hpa I, this promoter is utilized approximately 10-fold more efficiently than the other class II promoters. The mechanism of this activation is not yet known.


Terminally redundant deletion mutants of bacteriophage BF23. Deletion mutants of bacteriophage BF23 were isolated and the positions of the deletions were determined. Two different deletable regions were detected: one in the same region as previously reported for bacteriophage T5, which is closely related to BF23; and the other within both terminal repetitions. The former deletable region lay between positions 0.31 and 0.36, which represented the fractional lengths of the BF23 ( + ) DNA as measured from its left end. The latter deletion was evenly divided between the two terminal repetitions. The deletion in the left terminal repetition lay between positions 0.044 and 0.078 and was repeated in the corresponding region of the right terminal repetition between positions 0.966 and 1.0. The size of the DNA transferred to host cells during the first step of DNA transfer by BF23 carrying deletions in the terminal repetitions of its DNA was less than the size of DNA transferred during the first step by wild-type BF23 by an amount equal to the size of the deletion in each terminal repetition. This finding suggests the existence of a specific mechanism for delineating the position at which the first step of DNA transfer is stopped.


Influenza B virus genome: assignment of viral polypeptides to RNA segments. It was shown that all eight RNA segments of influenza B viruses are most likely monocistronic and code for eight virus-specific polypeptides. A genetic map of the influenza B virus genome was established, and six polypeptides (P1 protein, nucleoprotein, hemagglutinin, neuraminidase, M protein, and nonstructural protein) were unambiguously assigned to specific RNA segments. Molecular weight estimates of the eight individual genes are obtained by using the glyoxal method. These results suggest that each influenza B virus RNA segment has a greater molecular weight than the influenza A virus RNA segment which codes for the analogous gene product.


Physical characterization of the superhelical DNA genome of an enveloped mycoplasmavirus. Mycoplasmavirus MVL2 is a nonlytic enveloped virion containing DNA. This DNA has been shown to be a double-stranded circular superhelical molecule of 11.8 kilobase pairs (7.8 X 10(6) daltons). The superhelix density is greater than that of phi X174 RFI but less than that of PM2 phage DNA. A physical map of the MVL2 genome has been obtained using restriction endonucleases.


Characterization of mycoplasmavirus MVL2 DNA. The size and molecular configuration of mycoplasmavirus MVL2 DNA are presented. The DNA was shown to be a covalently closed circular duplex molecule with a molecular weight of 7.4 X 10(6). In sucrose gradients at neutral pH, the form I and form II DNA molecules have sedimentation values of approximately 29S and 23S, respectively. Under alkaline conditions, the form I and form II have S values of 70 and 20, respectively.


Subcellular distribution of newly synthesized virus-specific polypeptides in Moloney murine leukemia virus infected cells. Immune precipitation analysis of pulse-labeled proteins present in subcellular fractions of mouse embryo cells infected with Moloney murine leukemia virus showed the presence of anti-gp70 serum-precipitable viral envelope gene products mainly in the microsomal fractions of these cells. In contrast, anti-p30 serum-specific gag (group specific antigen) gene products were found to be distributed in similar amounts in both the microsomal and postmicrosomal supernatant fractions of pulse-labeled cells.


Inhibition of coronavirus 229E replication by actinomycin D. The yields of human coronavirus 229E grown in L132 cells were markedly inhibited by actinomycin D, the 50% inhibitory dose being 0.1 micron/ml. Inhibition was maximal during the early phase of virus replication, did not appear to involve viral RNA synthesis per se, and was shown to be dependent on the input multiplicity of infection.


Inhibition of vaccinia virus growth by zinc ions: effects on early RNA and thymidine kinase synthesis. Accumulation of thymidine kinase activity in vaccinia virus-infected cells was severely inhibited by zinc ions if the drug was added within 1 h postinfection. If added later, zinc ions had no effect on the enzyme synthesis. A fraction of RNA which is normally synthesized in infected cells, was missing from a proper part of the gradient if the cells were treated with zinc ions within 1 h postinfection (as has been shown by cosedimentation of pulse-labeled RNAs in isokinetic gradients). It is suggested that a transcriptional (or posttranscriptional) step is involved in zinc-caused inhibition of vaccinia virus growth.


Inhibition of entry of the lipid-containing bacteriophage PR4 by fatty acid derivatives. Amphiphilic fatty acid derivatives having a 14- to 20-carbon cis-unsaturated alkyl chain were found to be potent inhibitors of the entry process of the lipid-containing bacteriophage PR4.


Effects of 17beta-estradiol on the replication of rubella virus in an estrogen-responsive, continuous cell line. A 70% reduction in the number of infectious rubella virions normally produced in MCF-7 cells occurred when the cells were stimulated with 17beta-estradiol. Analysis of virus-induced RNA indicated a delay in the conversion of replicative intermediate to virion RNA.


"sarc" sequence transcription in Moloney sarcoma virus-transformed nonproducer cell lines. (3)H-labeled complementary DNA(sarc), complementary to the murine sarcoma virus (MSV)-specific portion of the Moloney MSV (M-MSV) genome, was prepared. M-MSV-specific RNA was then quantitated in the cytoplasm of several M-MSV-transformed, non-virus-producing, clonal NIH 3T3 cell lines. These lines, designated 71 N clones 5, 6, and 3, have been characterized previously by the degree to which they exhibit transformation properties and transcribe Moloney murine leukemia virus-related RNA (S. Salzberg and M. Green, J. Virol. 13:1001-1004, 1974; N. Tsuchida and M. Green, J. Virol. 14:587-591, 1974). By the criteria of cell morphology and agglutination by concanavalin A, cells of clone 5 are highly transformed, cells of clone 6 are almost normal in the sense that they resemble the parent NIH 3T3 cells, and cells of clone 3 are phenotypically intermediate. In the present study, the amounts of cytoplasmic MSV-specific RNA correlated well with the relative degrees of transformation of the cell lines, varying over 35-fold between the least transformed (clone 6) and most transformed (clone 5) lines. Superinfection of either clone 5 or clone 6 with Moloney murine leukemia virus resulted in a fivefold increase in the MSV-specific RNA in the cell cytoplasm. Evidence from (3)H-labeled complementary DNA:cell DNA hybridization studies indicated that the quantity of M-MSV-specific RNA in the nonproducer lines was not directly related to DNA provirus copy number in the cell DNA. Although clones 5 and 6 differ greatly in transformation characteristics and in MSV-specific RNA content, they each apparently contain about two copies of MSV-specific DNA sequence per haploid genome. Thus, factors such as site of provirus integration may be of primary importance in determining virus-specific transcription and cell transformation.


Recombination of bacteriophage phi X174 by the red function of bacteriophage lambda. Recombination of bacteriophage phi X174 was effectively promoted when the Red function of lambda was supplied by either co-infection with lambda or induction of lambda lysogens. Mutations in red alpha and red beta genes of lambda abolished recombination nearly completely, whereas a mutation in gam gene reduced it only slightly. The Red-promoted recombination of phi X174 occurred in recA, recB, and polA mutants as well as in wild-type strains of Escherichia coli. It was further stimulated when phi X174 mutants were irradiated with UV light before infection.


Autoregulation of adenovirus type 5 early gene expression. III. Transcription studies in isolated nuclei. The rate of adenovirus RNA synthesis was compared in nuclei isolated from cells infected at 40.5 degrees C in the presence of 1-beta-d-arabinofuranosylcytosine with adenovirus 5 or an early temperature-sensitive mutant of adenovirus type 5, H5ts125 (ts125). In nuclei isolated at various times after infection, the maximum amount of virus RNA synthesis occurred at 6 h after infection, after which time virus RNA synthesis declined in nuclei from wild-type infections but remained high in nuclei from ts125 infections. At 12 h after infection, the amount of virus RNA synthesis was 8- to 11-fold higher in nuclei from ts125 infections than in nuclei from wild-type infections. However, the kinetics of virus RNA synthesis in nuclei isolated from both infections were similar. When a ts125-infected culture was shifted to 32 degrees C for 3 h (12 to 15 h after infection) before nucleus isolation, the amount of virus RNA synthesis in the isolated nuclei was reduced to nearly wild-type levels. A pulse-chase experiment showed little difference in degradation rates of virus RNA in isolated nuclei from wild-type and ts125 infections. Hybridization of RNA synthesized in vitro to restriction fragments of adenovirus type 5 DNA was consistent with early virus RNA. These results support the idea that the 72,000-dalton DNA-binding protein encoded by the mutant gene in ts125 can regulate early adenovirus gene expression by inhibiting initiation of transcription of the adenovirus genome.


Carbohydrate structure of Sindbis virus glycoprotein E2 from virus grown in hamster and chicken cells. Sindbis virus was used as a probe to examine glycosylation processes in two different species of cultured cells. Parallel studies were carried out analyzing the carbohydrate added to Sindbis glycoprotein E2 when the virus was grown in chicken embryo cells and BHK cells. The Pronase glycopeptides of Sindbis glycoprotein E2 were purified by a combination of ion-exchange and gel filtration chromatography. Four glycopeptides were resolved, ranging in molecular weight from 1,800 to 2,700. Structures are proposed for each of the four glycopeptides, based on data obtained by quantitative composition analyses, methylation analyses, and degradation of the glycopeptides using purified exo- and endoglycosidases. The largest three glycopeptides (S1, S2, and S3) have similar structures but differ in the extent of sialylation. All three contain N-acetylglucosamine, mannose, galactose, and fucose, in a structure similar to oligosaccharides found on other glycoproteins. Glycopeptide S1 has two residues of sialic acid, whereas glycopeptides S2 and S3 contain 1 and 0 residues of sialic acid, respectively. The smallest glycopeptide, S4, contains only N-acetyglucosamine and mannose, and is also similar to mannose-rich oligosaccharides found on other glycoproteins. Each of the complex glycopeptides (S1, S2, or S3) from virus grown in BHK cells is indistinguishable from the corresponding glycopeptides derived from virus grown in chicken cells. Glycopeptide S4 is also very similar in size, composition, and sugar linkages from virus derived from the two hosts. These results suggest that chicken cells and BHK cells have similar glycosylation mechanisms and glycosylate Sindbis glycoprotein E2 in nearly identical ways.


Interruption-deficient mutants of bacteriophage T5 I. Isolation and general properties. Mutations of bacteriophage T5 were isolated which lack one or more of the natural single-chain interruptions that occur in the mature DNA of this virus. Interruption-deficient mutants were detected by screening survivors of hydroxylamine mutagenesis for altered DNA structure by electrophoresis in agarose slab gels. Over 60 independent mutants were isolated from a survey of approximately 800 phages particles. All of the mutants were viable and could be grouped into two classes. Mutants in one class lacked one of the localized sites where interruptions occur in T5 DNA. To date, mutants that affect five different sites have been obtained. Mutants in the other class were essentially free from interruptions or had a reduced frequency of interruptions throughout the genome. The members of this class included several amber mutants. Complementation tests indicated that at least two genes are required for the presence of interruptions in mature T5 DNA.


Interruption-deficient mutants of bacteriophage T5. II. Properties of a mutant lacking a specific interruption. An examination was made of the properties of T5HA4, a mutant of bacteriophage T5 that lacks the single-chain interruption that occurs at 7.9% from the left end of the genome. The DNAs of T5HA4 and the wild type were compared by electrophoresis in agarose gels of both single-stranded fragments produced by denaturation and duplex fragments generated by sequential treatment with exonuclease III and SI nuclease. These studies demonstrated that T5HA4 also lacks an interruption that occurs at 99.6% in wild-type DNA. The interruptions at 7.9 and 99.6% therefore occur within the 8.3% of T5 DNA that is terminally repetitious. Evidence on the location of other interruptions within the terminal repetition was also obtained. Analysis of T5HA4 with a restriction endonuclease indicated that the interruption deficiency is not due to a deletion or addition mutation. The injection of T5HA4 DNA into a host bacterium was found to occur, as with the wild type, in a two-step manner. The interruption at 7.9% is therefore not required for stopping DNA transfer after the initial 8% segment has been injected.


Structure of the murine leukemia virus envelope glycoprotein precursor. The glycosylated env gene precurosr (Pr80env) of Moloney murine leukemia virus has been isolated by selective immunoprecipitation. Use of the drug tunicamycin to inhibit nascent glycosylation or specific cleavage with endoglycosidase H demonstrated that the precursor contained an apoprotein with a molecular weight of 60,000. The finished virion glycoprotein (gp70) was largely resistant to the action of endoglycosidase H. Chromatography of the glycopeptides of Pr80env in conjunction with endoglycosidase H digestion studies suggested that the precursor contained two distinct major glycosylation sites. Analysis of partial proteolytic cleavage fragments of Pr80env before and after endoglycosidase H treatment placed the two glycosylation sites within a 30,000-dalton region of the apoprotein sequence. Kinetic experiments showed that carbohydrate processing as well as proteolytic cleavage are late steps in the maturation of Pr80env.


Nuclear accumulation of influenza viral RNA transcripts and the effects of cycloheximide, actinomycin D, and alpha-amanitin. The use of virus-specific (32)P-labeled complementary DNA and (125)I-labeled virion RNA as hybridization probes has allowed us to quantitate the number of molecules of complementary RNA (cRNA) and progeny virion RNA in MDCK cells infected with influenza virus. We compared the distribution of cRNA between the nucleus and the cytoplasm in cycloheximide-treated cells to that found in untreated cells, beginning 1 h after infection. A greater percentage of the total cRNA was detected in the nucleus of the drug-treated cells at all times investigated. For the first 2 h after infection about 50% of the cRNA synthesized in the cycloheximide-treated cells was found in the nucleus. These nuclear cRNA molecules were characterized and shown to be polyadenylated transcripts of each of the genome virion RNA segments. Viral cRNA synthesis was not completely inhibited by the addition of actinomycin D at the beginning of infection, with or without the concomitant addition of cycloheximide. A large fraction (about 90%) of these cRNA sequences were detected in the nucleus. Characterization of these nuclear cRNA molecules showed that they contained polyadenylic acid and represented transcripts of both those segments coding for proteins synthesized predominantly early after infection ("early" proteins) and those virion RNA segments coding for "late" proteins. Also, in vitro translation of these cRNA molecules showed that they were functional virus mRNA's. In contrast to actinomycin D, alpha-amanitin completely inhibited cRNA synthesis when added at the beginning of infection, and addition of this drug after 1.5 h had no effect on further cRNA synthesis.


Novel bacteriophage lambda mutation affecting lambda head assembly. A novel phage lambda mutation, called dc10, which interferes with proper lambda head assembly has been isolated and characterized. Phage lambda carrying this mutation is (i) unable to form plaques at 30 or 37 degrees C but does so at 42 degrees C and (ii) unable to form plaques at 42 degrees C on pN-constitutive hosts. Both properties are due to dc10 since all phage revertants for one phenotype simultaneously lose the other phenotype and vice versa. The dc10 mutation has been mapped in the B gene and has been shown to be dominant over the corresponding wild-type product. At 30 degrees C the dc10 mutation results in the formation of abnormal petit lambda heads made up of pE, pB, pC, and pNu3. Under pN-constitutive conditions, the dc10 mutation results in the formation of abnormal petit lambda heads made of pE, X1, and X2 only. A model to explain the data is presented.


Oligonucleotide mapping studies of standard and defective Sindbis virus RNA. Oligonucleotide mapping studies of the RNA from standard and defective interfering particles of Sindbis virus demonstrate that 3'- and 5'-terminal regions of the genome are conserved in the defective RNAs. These studies also suggest that defective RNAs contain multiple deletions.


Quantitative measurement of intracellular RNA genomes of Rauscher murine leukemia virus by competition hybridization in DNA excess. The technique of competition hybridization in DNA excess was used to determine the intracellular distribution of RNA genomes of Rauscher murine leukemia virus. An examination of subcellular RNA fractions revealed that 59% of intracellular viral RNA genomes were associated with the nuclear-enriched fraction, 41% with the cytoplasmic fraction, and 18% with the polysomal-enriched fraction. Also, an analysis of total cellular RNA disclosed that 20% of intracellular viral RNA genomes were polyadenylated and 80% were not polyadenylated.


Parental adenovirus type 2 genomes recovered early or late in infection possess terminal proteins. At both early (3 h) and late (18 h) times after infection of KB cells with adenovirus 2, more than 90% of parental nuclear viral genomes exist as complexes which contain terminally linked proteins. Density shift experiments employing 5-bromo-2'-deoxyuridine indicate that these parental DNA molecules remain complexed with terminal proteins after DNA replication. The persistent linkage of proteins to the termini of intranuclear viral DNA suggests that these proteins have an essential role in adenovirus replication.


A new embryologic classification for uro-gynecologic malformations: the syndromes of mesonephric duct induced müllerian deformities. A new embryologic classification is proposed to explain the association of unilateral renal agenesis with abnormalities of the female reproductive tract. An awareness of the syndrome of combined müllerian and mesonephric duct abnormalities may permit accurate anatomic diagnosis and spare patients needless surgical exploration.


Stent versus nephrostomy: is there a choice? When temporary or palliative urinary diversion is necessary the Gibbons ureteral stent has reduced markedly the need for nephrostomy. Of 48 patients 41 (85 per cent) had successful insertion of stents. Only 1 of these patients had to undergo nephrostomy at a later date. Complications have been minimal and the patients' tolerance has been excellent.


Retroperitoneal drainage of ureterointestinal conduits. Records of 132 patients undergoing ileal or colonic conduit urinary diversion were reviewed. Drains were placed retroperitoneally in all cases. Urinary leakage occurred most frequently in patients with malignancies and was associated with previous pelvic irradiation or simultaneous cystectomy. Most leaks closed spontaneously with non-operative management.


Primary non-urachal adenocarcinoma of the bladder. The clinical course of 34 patients with non-urachal adenocarcinoma of the bladder was reviewed and compared to the world experience. The 5-year survivorship was 19 per cent, with only 1 patient being free of disease. Metaplasia with formation of glandular elements reflects the unstable potential of the transitional cell and may be associated with the biologic aggressiveness of the tumor.


Transurethral laser urethrotomy in man: preliminary report. After in vitro and in vivo endoscopic laser investigation on normal and experimentally strictured urethras in dogs transurethral laser urethrotomy was done on men. A neodymium: yttrium aluminum garnet laser with a flexible laser light guide was used. The endoscope developed for this purpose is described in detail. The first laser operations were without complications.


Experience with penile prosthesis in spinal cord injury patients. During the last 2 years 30 patients in the spinal cord injury service at our hospital received a penile prosthesis implant. The records of 25 patients were reviewed and 20 patients were interviewed 3 to 27 months postoperatively. A detailed evaluation of each patient shows that in properly selected patients the results of this operation can be satisfactory and gratifying.


Prostatic secretion leukocyte studies in non-bacterial prostatitis (prostatosis). Non-bacterial prostatitis is a common affliction among men and lacks objective criteria for clear identification. We studied 43 consecutive patients and 20 normal controls for the presence of bacteria and quantitation of prostatic secretion leukocytes. Significantly greater numbers of macrophages per volume of prostatic secretion were found in the patients. Multiple observations of such increased leukocyte proliferation could be used to establish true prostatic inflammation.


The impact of current staging procedures in assessing disease extent of prostatic adenocarcinoma. We studied 454 patients with prostatic adenocarcinoma who were assigned a preliminary clinical stage on the basis of serum acid phosphatase, routine bone survey and physical examination. Subsequently, they were assigned a final clinical stage after radioisotopic bone scanning, lymphangiography and staging pelvic lymph node dissection. Only 53, 54, 57 and 26 per cent, respectively, of patients initially assigned the preliminary clinical stage of IB, II, III or IVA remained at that stage after the additional studies.


Results of biopsy after early stage prostatic cancer treatment by implantation of 125I seeds. We have treated 77 patients for clinically early stage carcinoma of the prostate, 9 stage A2, 63 stage B and 5 stage C, with direct implantation of 125I seeds into the prostate and pelvic lymphadenectomy. It is estimated that a minimum dose of 15,000 rad but a maximum dose of 35,000 rad is delivered to the prostate over several months. Of the 77 patients 14 (18 per cent) had metastatic disease in the pelvic lymph nodes. In 22 cases perineal needle biopsy was done 12 to 18 months after treatment and in 3 cases a second biopsy was performed after 2 to 3 years. Persistent tumor was present in 11 biopsies. Cytological changes were observed in 8 of these, primarily cytoplasmic vacuolation and nuclear pyknosis. There seemed to be no relationship between grade and stage of disease and histological evidence of persistence of tumor after radiation. One patient with persistent tumor in the postoperative biopsy has shown progression of disease after 2 years and another with a negative biopsy has a bony metastasis. The remaining 10 patients with persistent tumor have shown no sign of progression of disease during a 2 to 4-year interval.


Spontaneous remission of chyluria.

Spontaneous remission of chyluria. The natural history of chyluria is not well known. We have followed 72 patients with chyluria who have not had treatment and the condition has disappeared in 36 cases (50 per cent). In these cases the duration of chyluria varied from 3 days to 20 years, with an average of 44.3 months, but it was less than 6 months in about half of the cases.


Weather and the incidence of urinary stone colic. There were 204 patients in one specific district with renal colic who were studied to determine the relationship of the condition to climatic conditions. It was found that stones were prone to cause colic on warm days when air pressure was falling.


Ultrasound-guided percutaneous nephrostomy in the pediatric patient. Percutaneous nephrostomy drainage may be established rapidly using ultrasonic guidance in severely ill pediatric patients with obstructive uropathy. B-mode ultrasonography has proved to be rapid, accurate and free of irradiation hazards, making it applicable to children with little or no sedation.


Salvaging the iatrogenic megaureter.

Salvaging the iatrogenic megaureter. From 1965 through 1975 we treated 30 children with 44 megaureters that were acquired after the reimplantation of normal caliber ureters. One-third of these children had had at least 2 prior surgical procedures on the bladder and 30 per cent were azotemic. There was no difference in the salvage rate of acquired megaureters with reflux versus obstruction. Temporary proximal diversion often was useful in the short-term management. Only 70 per cent of the patients completed reconstruction and only 70 per cent of those reconstructed were successful by all parameters. Ureteral tailoring was generally a necessary adjunct to successful repeat reimplantation. There was a significantly lesser salvage rate in those ureters that had had more than 1 prior surgical procedure. The best results were obtained when all surgical maneuvers were done transvesically because there was a significant incidence of acquired atonic vesical dysfunction after repeated dissections in the perivesical area.


Hypothermia during infant cystoscopy.

Hypothermia during infant cystoscopy. Two cases of hypothermia during prolonged infancy cystoscopy are reported. A significant decrease in the core temperature also was noted when irrigating the bladders of small mongrel dogs with room temperature water. The complications of infant hypothermia are reviewed and the factors that predispose the infant to acute changes in body temperature are emphasized. The measures of maintaining the core temperature in the pediatric surgical patient are discussed and particular situations when prolonged cystoscopy may occur, necessitating warming of irrigation fluids, are presented.


Cecil urethroplasty with concurrent scrotoplasty for repair of hypospadias. During 13 years about 8 per cent of 234 patients operated upon for hypospadias presented surgical problems of reconstruction of the urethra from the urethral meatus proximal to the penoscrotal junction. These cases commonly are referred to as perineal or penoscrotal hypospadias. The problem occurs when the urethral groove fails to develop adequately and the scrotal folds have failed to rotate caudally. This hypospadias is in contrast to penoscrotal or distal hypospadias, when the urethral groove is developed but fusion is incomplete. In these patients the Cecil procedure has been used but modified to extend the urethra from behind the penoscrotal junction to the glans at the second stage of the urethroplasty. The modification consists of rotating the scrotum caudally from its high location. The tension and acute angulation that contributed to frequent complications previously associated with such attempts have been avoided and results are satisfactory.


Delayed development of the scrotum in exstrophy. Early attempts at orchiopexy should be avoided in children with exstrophy, since the testes will often descend spontaneously by puberty. In many cases the scrotum will grow much larger and will become pendulous by puberty. In a patient with a rudimentary scrotum orchiopexy will only result in fixing the testes in an abnormally high position owing to the lack of a sac into which to place them. If the scrotum develops later the testicles will probably be unable to descend further. Scrotal skin should not be used for urethroplasty operations in children with exstrophy and special techniques should be applied if it is necessary to repair hernias in these patients. In the child with partially descended testes but with only a small patch of wrinkled scrotum efforts may be undertaken to increase scrotal size by topical hormone treatment (or by injections) before the onset of puberty to help fertility and emotional development.


Mixed gonadal dysgenesis.

Mixed gonadal dysgenesis. Mixed gonadal dysgenesis is an intersex syndrome characterized by a unilateral streak gonad, persistent müllerian duct structures and ambiguous genitalia. These patients are chromatin negative and exhibit XO/XY mosaicism. The disorder probably results from a cytogenetic error that occurs early in embryogenesis. These patients usually are reared as female subjects but masculinity begins at puberty because of the qualitatively and quantitatively normal testicular androgenic function. The patients are diagnosed on the basis of a Y chromosomal component and the presence of a unilateral testis and contralateral streak gonad at the time of exploratory laparotomy. The testis and the streak gonad should be removed because of the potential development of a gonadoblastoma and the virilization that occurs at puberty.


Ectopic adrenal in inguinal hernia sacs. Nodules of the ectopic adrenal cortex were identified in hernia sacs in 5 men. This condition seems to result from developing adrenal cortical tissue accompanying a descending testis. Ectopic adrenal must be distinguished from metastatic clear cell carcinoma. Such ectopic foci could potentially result in neoplasms or be a source of hormone production after adrenalectomy.


Percutaneous U-loop nephrostomy.

Percutaneous U-loop nephrostomy. Single-tube nephrostomy drainage can be converted to U-loop drainage using percutaneous methods. Because only local anesthesia is required the technique can be used for patients who are poor operative risks.


Traumatic caliceal-parenchymal disruption. A unique case is described of traumatic disruption of an intact collecting system (calices, renal pelvis and ureter) from an intact kidney. Evidence is presented for a congenital weakness at the level of disruption.


Chemotherapy of advanced transitional cell carcinoma of the renal pelvis: report of 3 cases treated with vinblastine and chloroethyl-cyclohexyl-nitrosourea. Three patients with advanced transitional cell carcinoma of the renal pelvis were treated with 5 mg./m.2 body surface vinblastine intravenously weekly and 130 mg./m.2 chloroethyl-cyclohexyl-nitrosourea orally with 6-week intervals. One patient had a partial remission for 1.5 months and 2 had stationary disease for 6 and 2 months.


Ectopic single ureter and severe renal dysplasia: an unusual presentation. An ectopic single ureter in the male subject is a rare anomaly, which commonly drains a dysplastic kidney to the prostatic urethra or the seminal vesicle. The further the ureteral orifice is located from the trigone of the bladder, the more severe is the ipsilateral renal dysplasia. A case is described in which an ectopic ureter and ectopic and dysplastic kidney were found at the time of inguinal herniorrhaphy. The ureter was connected to the epididymis of the testicle. This is the most severe form of ureteral ectopia and it has not been reported previously.


Leiomyoma of the female urethra.

Leiomyoma of the female urethra. Leiomyoma of the urethra is a rare condition, which is more common in female than in male subjects. There are only 14 documented cases of leiomyoma of the female urethra in the literature. A careful analysis of these cases revealed the average patient age to be 34.1 years. The site of the lesion determined the clinical features and the surgical approach. No common etiological factor could be ascertained. Two additional cases of leiomyoma of the female urethra are reported.


Non-neoplastic cystic lesions of the tunica albuginea: an electron microscopic and clinical study of 2 cases. Cystic lesions of the tunica albuginea are rare and of uncertain histogenesis, and may present diagnostic difficulties. Recently, we encountered 2 such lesions that we were able to study by electron microscopy. We report the cases and present the electron microscopic observations that suggest the histogenesis of these lesions.


Malignant paraganglioma of the prostate and retroperitoneum. A case of malignant paraganglioma of the prostate, metastatic to the retroperitoneal area, is reported. The literature is reviewed and the possible origin of the disease is discussed. We believe that this is the first report of this entity of prostatic origin.


Anterior bladder tube flap reconstruction of the urethrovesical neck after radical retropubic prostatectomy. Difficulty with urethrovesical neck anastomosis after radical retropubic prostatectomy led us to form an anterior bladder tube flap for anastomosis to the transected urethra in 5 selected cases. We found that combining the anterior bladder tube flap technique with radical retropubic prostatectomy facilitates the urethrovesical neck anastomosis and improves the transient postoperative incontinence sometimes encountered. Results of the 5 patients in whom this technique was used form the basis for a brief discussion of the technique.


Postvaccine era measles epidemiology.

Postvaccine era measles epidemiology. Measles incidence in the United States was calculated from estimates of the United States Immunization Survey for the years 1965 to 1975. These data were examined according to birth cohorts and cross-sectionally by age to determine the effect of live measles virus vaccine on measles epidemiology. The results showed a high rate of infection for children aged 1 year, followed by a decrease in the infection rates for preschool children, with rates increasing agin at ages 5 and 6 years. It was hypothesized that an effective vaccine-induced herb immunity exists in preschool children until they are of school age. When incidence rates were based on susceptible children, the highest rates occurred in the school-aged populations.


The National Football Head and Neck Injury Registry. Report and conclusions 1978. The National Football Head and Neck Injury Registry has documented 1,129 injuries since 1971 that involved hospitalization for more than 72 hours, surgical intervention, fracture-dislocation, permanent paralysis, or death. Of this group of injuries, 550 were fracture-dislocations of the cervical spine, of which 176 were associated with permanent quadriplegia. It appears that during the last two decades, there has been a decrease in the incidence of direct fatalities, head injuries associated with intracranial hemorrhage, and injuries associated with death. Conversely, cervical spine injuries with fracture-dislocation and with permanent quadriplegia have increased. We believe that these observations are the result of the development of a protective helmet-face mask system that has effectively protected the head, and by so doing has allowed it to be used as a battering ram in tackling and blocking techniques, thus placing the cervical spine at risk of injury.


Upper trunk brachial plexopathy in football players. Ten football players seen from 1973 through 1977 at the University of Wisconsin Hospitals were found to have clinical and electrodiagnostic evidence of injury to the upper trunk of the brachial plexus. Each had upper limb paresis following one or more blows to the head or shoulders. The development of persistent weakness often was preceded by burning paresthesias in the upper limb. Our experience suggests that the syndrome of burning paresthesias and subsequent arm weakness frequently is secondary to stretching of the brachial plexus.